Immunoconjugates targeting cd46 and methods of use thereof

Abstract

This article discloses an immune conjugate comprising a CD46 binding domain and an effector. This article further provides a method for treating cancer comprising administering to an individual with cancer a pharmaceutical composition comprising an immune conjugate containing a CD46 binding domain and an effector.

Description

Prior Technology

[0001] CD46, also known as CD46 complement regulatory protein, differentiation cluster 46, and membrane cofactor protein, is an inhibitory complement receptor. Overexpression of CD46 has been observed in several cancers, such as breast cancer, colorectal cancer, liver cancer, lung cancer, and prostate cancer. In some cases, CD46 overexpression is a characteristic of negative prognostic factors. For example, CD46 overexpression has been associated with shorter progression-free survival and shorter overall survival in breast cancer and ovarian cancer patients. New targeted therapies and treatment regimens for cancer treatment are needed. Summary of the Invention

[0002] This invention provides an immune conjugate for treating conditions characterized by CD46 expression on the cell surface, such as metastatic castration-resistant prostate cancer and multiple myeloma.

[0003] In some embodiments, the immune conjugate is present at concentrations of approximately 1.0 to 4.5 mg / kg, approximately 1.0 to 4.0 mg / kg, approximately 1.0 to 3.5 mg / kg, approximately 1.0 to 3.0 mg / kg, approximately 1.0 to 2.57 mg / kg, approximately 1.0 to 2.5 mg / kg, approximately 1.0 to 2.4 mg / kg, approximately 1.5 to 4.5 mg / kg, approximately 1.5 to 4.0 mg / kg, approximately 1.5 to 3.5 mg / kg, approximately 1.5 to 3.0 mg / kg, approximately 1.5 to 2.57 mg / kg, approximately 1.5 to 2.5 mg / kg, approximately 1.5 to 2.5 mg / kg, approximately 1.5 to 2.4 mg / kg, approximately 1.5 to 2.0 mg / kg, approximately 1.8 to 4.5 mg / kg, approximately 1.8 to 4.0 mg / kg, approximately 1.8 to 3.5 mg / kg, approximately 1.8 to 3.0 mg / kg, and approximately 1.8 to 2.57 mg / kg. The immune conjugate was administered to the human individual at a dose of about 1.8 to about 2.5 mg / kg, about 1.8 to about 2.4 mg / kg, or about 1.8 to about 2.0 mg / kg. In some embodiments, the immune conjugate was administered to the human individual at a dose of about 1.5 to about 2.5 mg / kg. In some embodiments, the immune conjugate is administered to the human individual at a dose of about 1.0, about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2.0, about 2.1, about 2.2, about 2.3, about 2.4, about 2.5, about 2.6, about 2.7, about 2.8, about 2.9, about 3.0, about 3.1, about 3.2, about 3.3, about 3.4, about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, or about 4.0 mg / kg. In some embodiments, the immune conjugate is administered to the human individual at a dose of about 1.8, about 2.4, or about 3.2 mg / kg. In some embodiments, the immune conjugate is administered to the human individual at a dose of about 1.8 mg / kg. In some embodiments, the immune conjugate is administered to the human individual at a dose of about 2.4 mg / kg. In some embodiments, the immune conjugate is administered to the human individual at a dose of about 3.2 mg / kg.

[0004] In some embodiments, the immune conjugate is administered to the human individual via intravenous infusion. In some embodiments, the immune conjugate is administered to the human individual every 7 days, every 14 days, every 18 days, every 21 days, or every 30 days. In some embodiments, the immune conjugate is administered to the human individual every 21 days.

[0005] In some embodiments, the recombinant antibody system is bound to an effector, wherein the effector comprises a drug (or a prodrug thereof), a peptide, a protein, a detectable label, a liposome containing the drug (or a prodrug thereof), a radioactive nucleus, a viral particle, or a chelating agent. In some embodiments, the effector comprises a drug. In some embodiments, the drug is an anticancer drug. In some embodiments, the drug is a chemotherapeutic agent. In some embodiments, the drug is a microtubule inhibitor, a DNA disruptor, or a polymerase inhibitor. In some embodiments, the drug is a microtubule inhibitor. In some embodiments, the microtubule inhibitor is auristatin (or a derivative thereof), dolastatin-10 (or a derivative thereof), or maytansine (or a derivative thereof). In some embodiments, the microtubule inhibitor is monomethylauristatin F (MMAF), auristatin E (AE), monomethylauristatin E (MMAE), valine-citrulline MMAE (vcMMAE), or valine-citrulline MMAF (vcMMAF). In some embodiments, the microtubule inhibitor is monomethylauratestatin E (MMAE).

[0006] In some embodiments, the ratio of the effector to the recombinant antibody is about 3 to about 5. In some embodiments, the ratio of the effector to the recombinant antibody is about 4.

[0007] In some embodiments, the effector is bound to the recombinant antibody via a linker. In some embodiments, the linker is a peptide linker, a small molecule linker, or a linker comprising a peptide and a small molecule. In some embodiments, the linker comprises maleiminohexylacetyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB).

[0008] In some embodiments, the cancer is prostate cancer. In some embodiments, the prostate cancer is castration-resistant prostate cancer. In some embodiments, the cancer is multiple myeloma. In some embodiments, the multiple myeloma is relapsed or refractory multiple myeloma.

[0009] In some embodiments, the immune conjugate binds to CD46 expressed on the cell surface and is internalized into the cell. In some embodiments, the immune conjugate is internalized into the cell via macropinocytosis.

[0010] In another embodiment, the present invention provides a method for treating cancer in a human individual in need, the method comprising administering to the individual an immune conjugate comprising: (a) a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region including three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3, and a light chain (LC) variable region including three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 respectively comprise the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, and LC CDR1, LC CDR2, and LC CDR3 respectively comprise the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively. (a) the amino acid sequence of 6; and (b) monomethyl auristatin E (MMAE) bound to the recombinant antibody via a maleiminohexyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB) linker; wherein the immunoconjugate is administered at a dose of about 1.0 to about 4.0 mg / kg.

[0011] In another embodiment, the present invention provides a pharmaceutical composition comprising: (a) an immunoconjugate at a concentration of about 10.0 ± 5.0 mg / mL, and (b) a histidine buffer; wherein the immunoconjugate comprises: (a) a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region comprising three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3, and a light chain (LC) variable region comprising three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 comprise the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and LC CDR1, LC CDR2, and LC CDR3 comprise the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively; and (b) an effector that binds to the recombinant antibody.

[0012] In some embodiments, the pharmaceutical composition comprises about 10 to about 30 mM histidine buffer. In some embodiments, the pharmaceutical composition comprises about 10 to about 20 mM histidine buffer. In some embodiments, the pharmaceutical composition further comprises a cryoprotectant. In some embodiments, the cryoprotectant is a sugar. In some embodiments, the cryoprotectant is sucrose or trehalose. In some embodiments, the pharmaceutical composition further comprises a stabilizer. In some embodiments, the stabilizer prevents denaturation of the recombinant antibody, prevents aggregation of the immunoconjugates, or both. In some embodiments, the stabilizer is polysorbate. In some embodiments, the stabilizer is polysorbate 80. In some embodiments, the pharmaceutical composition has a pH of about 5.0 to about 7.0.

[0013] In some embodiments, the stabilizer is a polymer. In some embodiments, the polymer is a synthetic or semi-synthetic polymer. The polymer may be a linear polymer, such as polyvinylpyrrolidone or polyvinyl alcohol. The polymer may be a copolymer, such as a PVA-PEG graft copolymer. The polymer may be ionic, such as sodium carboxymethyl cellulose, sodium alginate, chitosan, or polyethylene glycol. The semi-synthetic polymer may be a nonionic polymer, such as HPMC, HPC, or HEC. In some embodiments, the stabilizer is a surfactant. The surfactant may be an ionic surfactant, such as sodium docusate, sodium lauryl sulfate, or polyethyleneimine, or a nonionic surfactant, such as Tween surfactants, poloxamer, D-α-tocopherol, polyethylene glycol succinate, or a block copolymer of polyethylene oxide-polyethylene oxide-polyethylene oxide. In some embodiments, the stabilizer is a food protein, amino acid, or copolymer. In some embodiments, the stabilizer is Captisol, Monosteol, microcrystalline cellulose and carboxymethyl cellulose, sorbitol, or cellulose gel.

[0014] In some embodiments, the pharmaceutical composition comprises a buffer. The buffer may be selected from acetates, citrates, tartrates, histidine, glutamic acid, phosphates, Tris, glycine, bicarbonates, succinates, sulfates, or nitrates. In some embodiments, the pharmaceutical composition comprises a tonic modifier. The tonic modifier may be selected from mannitol, sorbitol, lactose, dextrose, trehalose, sodium chloride, potassium chloride, glycerol, and glycerol. In some embodiments, the pharmaceutical composition comprises a buildup agent. The buildup agent may be a sugar or polyol selected from sucrose, trehalose, lactose, sorbitol, mannitol, and glycerol. The buildup agent may be an amino acid selected from arginine, aspartic acid, glutamic acid, lysine, proline, glycine, histidine, methionine, and alanine. The buildup agent may be a polymer or protein selected from gelatin, PVP, PLGA, PEG, dextran, cyclodextrins and their derivatives, starch derivatives, HAS, and BSA. In some embodiments, the pharmaceutical composition comprises an antioxidant. Antioxidants may be selected from histamine, methionine, ascorbic acid, glutathione, vitamin E, or poly(ethyleneimine). In some embodiments, the pharmaceutical composition includes an antimicrobial preservative. Pharmaceutical preservatives may be selected from benzyl alcohol, m-cresol, phenol, and 2-phenoxyethanol. In some embodiments, the pharmaceutical composition may include a chelating agent and / or a complexing agent. Chelating agents may be edetate disodium, diethylenetriamine pentaacetate, citric acid, hexaphosphate, thioglycolic acid, or zinc.

[0015] In some embodiments, the recombinant antibody system is bound to an effector, wherein the effector comprises a drug (or a prodrug thereof), a peptide, a protein, a detectable label, a liposome containing a drug (or a prodrug thereof), a radionuclides, a viral particle, or a chelating agent. In some embodiments, the effector comprises a drug. In some embodiments, the drug is an anticancer drug. In some embodiments, the drug is a chemotherapeutic agent. In some embodiments, the drug is a microtubule inhibitor, a DNA disruptor, or a polymerase inhibitor. In some embodiments, the drug is a microtubule inhibitor. In some embodiments, the microtubule inhibitor is aurestatin (or a derivative thereof), sulphoside-10 (or a derivative thereof), or maytansin (or a derivative thereof). In some embodiments, the microtubule inhibitor is monomethylaurestatin F (MMAF), aurestatin E (AE), monomethylaurestatin E (MMAE), valine-citrulline MMAE (vcMMAE), or valine-citrulline MMAF (vcMMAF). In some embodiments, the microtubule inhibitor is monomethylaurestatin E (MMAE). In some embodiments, the ratio of the effector to the recombinant antibody in the group of immune conjugates is about 3 to about 5. In some embodiments, the ratio of the effector to the recombinant antibody in the group of immune conjugates is approximately 4.

[0016] In some embodiments, the effector is bound to the recombinant antibody via a linker. In some embodiments, the linker is a peptide linker, a small molecule linker, or a linker comprising a peptide and a small molecule. In some embodiments, the linker comprises maleiminohexylacetyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB).

[0017] In another embodiment, the present invention provides a pharmaceutical composition comprising an immunoconjugate at a concentration of about 10.0 ± 1.0 mg / mL, about 20 mM histidine buffer, about 8.0% sucrose, and about 0.01% polysorbate 80; wherein the immunoconjugate comprises: (a) a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region comprising three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3, and a light chain (LC) variable region comprising three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 respectively comprise the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, and LC CDR1, LC CDR2, and LC CDR3 respectively comprise SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 1, SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively. (a) The amino acid sequence of 6; (b) Monomethyl auristatin E (MMAE) bound to the recombinant antibody via a maleiminohexyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB) linker.

[0018] In another embodiment, the present invention provides a method for treating relapsed or refractory multiple myeloma (RRMM) in a human individual in need, the method comprising administering to the individual a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region containing three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3, and a light chain (LC) variable region containing three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 respectively contain the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, each having 0 to 3 amino acid modifications, and LC CDR1, LC CDR2, and LC CDR3 respectively contain the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, each having 0 to 3 amino acid modifications.

[0019] In another embodiment, the present invention provides a method for treating relapsed or refractory multiple myeloma (RRMM) in a human individual in need, the method comprising administering to the individual a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region containing three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3, and a light chain (LC) variable region containing three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 contain the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and LC CDR1, LC CDR2, and LC CDR3 contain the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively.

[0020] In another embodiment, the present invention provides a method for treating metastatic castration-resistant prostate cancer in a human individual in need, the method comprising administering to the individual a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region containing three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3, and a light chain (LC) variable region containing three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 respectively contain the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, each having 0 to 3 amino acid modifications, and LC CDR1, LC CDR2, and LC CDR3 respectively contain the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, each having 0 to 3 amino acid modifications.

[0021] In another embodiment, the present invention provides a method for treating metastatic castration-resistant prostate cancer in a human individual in need, the method comprising administering to the individual a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region containing three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3, and a light chain (LC) variable region containing three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 contain the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and LC CDR1, LC CDR2, and LC CDR3 contain the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively.

[0022] In some embodiments, a recombinant antibody system for treating relapsed or refractory multiple myeloma or castration-resistant prostate cancer is combined with an effector, wherein the effector comprises a drug (or a prodrug thereof), a peptide, a protein, a detectable label, a liposome containing a drug (or a prodrug thereof), a radionuclides, viral particles, or a chelating agent. In some embodiments, the effector comprises a drug. In some embodiments, the drug is an anticancer drug. In some embodiments, the drug is a chemotherapeutic agent. In some embodiments, the drug is a microtubule inhibitor, a DNA disruptor, or a polymerase inhibitor. In some embodiments, the drug is a microtubule inhibitor. In some embodiments, the microtubule inhibitor is aurestatin (or a derivative thereof), sulphurin-10 (or a derivative thereof), or maytansin (or a derivative thereof). In some embodiments, the microtubule inhibitor is monomethylaurestatin F (MMAF), aurestatin E (AE), monomethylaurestatin E (MMAE), valine-citrulline MMAE (vcMMAE), or valine-citrulline MMAF (vcMMAF). In some embodiments, the microtubule inhibitor is monomethylaurestatin E (MMAE). In some embodiments, the ratio of the effector to the recombinant antibody is about 3 to about 5. In some embodiments, the ratio of the effector to the recombinant antibody is about 4.

[0023] In some embodiments, the effector of a method for treating relapsed or refractory multiple myeloma or castration-resistant prostate cancer is bound to the recombinant antibody via a linker. In some embodiments, the linker is a peptide linker, a small molecule linker, or a linker comprising a peptide and a small molecule. In some embodiments, the linker comprises maleimine hexacyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB).

[0024] In some embodiments, the recombinant antibiotic system used in a method for treating relapsed or refractory multiple myeloma or castration-resistant prostate cancer is expressed at concentrations of approximately 1.0 to approximately 4.5 mg / kg, approximately 1.0 to approximately 4.0 mg / kg, approximately 1.0 to approximately 3.5 mg / kg, approximately 1.0 to approximately 3.0 mg / kg, approximately 1.0 to approximately 2.7 mg / kg, approximately 1.0 to approximately 2.5 mg / kg, approximately 1.0 to approximately 2.4 mg / kg, approximately 1.5 to approximately 4.5 mg / kg, approximately 1.5 to approximately 4.0 mg / kg, approximately 1.5 to approximately 3.5 mg / kg, approximately 1.5 to approximately 3.0 mg / kg, approximately 1.5 to approximately 2.7 mg / kg, approximately 1.5 to approximately 2.5 mg / kg, approximately 1.5 to approximately 2.4 mg / kg, approximately 1.5 to approximately 2.0 mg / kg, approximately 1.8 to approximately 4.5 mg / kg, approximately 1.8 to approximately 4.0 mg / kg, and approximately 1.8 to approximately 3.5 mg / kg. The recombinant antibody system is administered at doses of approximately 1.8 to approximately 3.0 mg / kg, approximately 1.8 to approximately 2.57 mg / kg, approximately 1.8 to approximately 2.5 mg / kg, approximately 1.8 to approximately 2.4 mg / kg, or approximately 1.8 to approximately 2.0 mg / kg. In some embodiments, the recombinant antibody system is administered at a dose of approximately 1.5 to approximately 2.5 mg / kg. In some embodiments, the recombinant anti-inflammatory system is administered at a dose of about 1.0, about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2.0, about 2.1, about 2.2, about 2.3, about 2.4, about 2.5, about 2.6, about 2.7, about 2.8, about 2.9, about 3.0, about 3.1, about 3.2, about 3.3, about 3.4, about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, or about 4.0 mg / kg. In some embodiments, the recombinant anti-inflammatory system is administered at a dose of about 1.8, about 2.4, or about 3.2 mg / kg. In some embodiments, the recombinant anti-inflammatory system is administered at a dose of about 1.8 mg / kg. In some embodiments, the recombinant anti-inflammatory system is administered at a dose of about 2.4 mg / kg. In some embodiments, the recombinant antibody system is administered at a dose of approximately 3.2 mg / kg.

[0025] In some embodiments, the recombinant antibody system for treating relapsed or refractory multiple myeloma or castration-resistant prostate cancer is administered to the human individual via intravenous infusion. In some embodiments, the recombinant antibody system is administered to the human individual every 7 days, every 14 days, every 18 days, every 21 days, every 28 days, or monthly. In some embodiments, the recombinant antibody system is administered to the human individual every 21 days.

[0026] In another embodiment, the present invention provides a method for treating relapsed or refractory multiple myeloma (RRMM) in a human individual in need, the method comprising administering an immune conjugate to the individual, wherein the immune conjugate comprises (i) a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region comprising three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3, and a light chain comprising a light chain (LC) variable region comprising three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 comprise the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and LC CDR1, LC CDR2, and LC CDR3 comprise the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively; the antibody is conjugated via a linker to (ii) monomethylaurestatin E. (MMAE) binding, wherein the linker comprises maleiminohexylidene-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB).

[0027] In another embodiment, the present invention provides an immune conjugate comprising: a recombinant antibody comprising: a first heavy chain of SEQ ID NO: 9, a first light chain of SEQ ID NO: 10, a second heavy chain of SEQ ID NO: 9, and a second light chain of SEQ ID NO: 10; and one, two, three, or four adducts; wherein each of the one, two, three, or four adducts comprises monomethylauratestatin E conjugated to the recombinant antibody via a maleiminohexyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB) linker. (MMAE); wherein each of the first, second, third, or fourth adductor pairs binds to one pair of cysteine ​​residues of the recombinant antibody, wherein the cysteine ​​residue pairs are selected from: C219 of the first heavy chain and C214 of the first light chain; C219 of the second heavy chain and C214 of the second light chain; C225 of the first heavy chain and C225 of the second light chain; and C228 of the first heavy chain and C228 of the second light chain. In some embodiments, the immune conjugate comprises two pairs of such adductor pairs.

[0028] In another embodiment, the present invention provides a pharmaceutical composition comprising an immune conjugate comprising: a recombinant antibody comprising: a first heavy chain of SEQ ID NO: 9, a first light chain of SEQ ID NO: 10, a second heavy chain of SEQ ID NO: 9, and a second light chain of SEQ ID NO: 10; and one, two, three, or four adducts; wherein each of the one, two, three, or four adducts comprises monomethylauratestatin E conjugated to the recombinant antibody via a maleiminohexyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB) linker. (MMAE); wherein each of the first, second, third, or fourth adduct pairs binds to one pair of cysteine ​​residues of the recombinant antibody, wherein such cysteine ​​residue pairs are selected from: C219 of the first heavy chain and C214 of the first light chain; C219 of the second heavy chain and C214 of the second light chain; C225 of the first heavy chain and C225 of the second light chain; and C228 of the first heavy chain and C228 of the second light chain; the concentration of the immunoconjugate is approximately 10.0 ± 1.0 mg / mL, approximately 20 mM histidine buffer at pH 6.0, approximately 8.0% sucrose, and approximately 0.01% polysorbate 80.

[0029] In another embodiment, the present invention provides a pharmaceutical composition comprising: an immunoconjugate at a concentration of about 10.0 ± 1.0 mg / mL, about 20 mM histidine buffer, about 8.0% sucrose, and about 0.01% polysorbate 80; and wherein the immunoconjugate comprises: (a) a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region comprising three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3, and a light chain (LC) variable region comprising three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 respectively comprise the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, and LC CDR1, LC CDR2, and LC CDR3 respectively comprise SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 1, SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively. (a) the amino acid sequence of 6; and (b) monomethyl auristatin E (MMAE) bound to the recombinant antibody via a maleiminohexyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB) linker.

[0030] In another embodiment, the present invention provides a pharmaceutical composition comprising: an immune conjugate, a pharmaceutically acceptable buffer, and a pharmaceutically acceptable stabilizer; wherein the immune conjugate comprises (a) a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region comprising three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3, and a light chain (LC) variable region comprising three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 comprise the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and LC CDR1, LC CDR2, and LC CDR3 comprise the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively; and (b) an effector that binds to the recombinant antibody. In some embodiments, the pharmaceutical composition has a pH of about 5.0 to about 7.0. In some embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable buffer; wherein the buffer comprises citrate, phosphate, acetate, glycerol, histidine, succinate, malate, or α-ketoglutarate. In some embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable buffer; wherein the buffer comprises about 10 mM to about 30 mM histidine, and the pH is about 5 to about 7. In some embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable buffer; wherein the buffer comprises citrate, phosphate, acetate, glycerol, histidine, succinate, malate, or α-ketoglutarate; wherein the buffer comprises about 20 mM histidine, and the pH is about 6.0. In some embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable stabilizer; wherein the stabilizer prevents denaturation of the recombinant antibody, prevents aggregation of the immune conjugates, or both. In some embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable stabilizer; wherein the stabilizer comprises a nonionic surfactant. In some embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable stabilizer; wherein the stabilizer comprises polysorbate. In some embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable stabilizer; wherein the stabilizer comprises about 0.01% polysorbate-80. In some embodiments, a pharmaceutical composition comprising an immune binder, a pharmaceutically acceptable buffer, and a pharmaceutically acceptable stabilizer further comprises a pharmaceutically acceptable cryoprotectant. In some embodiments, a pharmaceutical composition comprising an immune binder, a pharmaceutically acceptable buffer, and a pharmaceutically acceptable stabilizer further comprises a pharmaceutically acceptable cryoprotectant; wherein the cryoprotectant comprises sugar.In some embodiments, the pharmaceutical composition comprising an immune binder, a pharmaceutically acceptable buffer, and a pharmaceutically acceptable stabilizer further comprises a pharmaceutically acceptable cryoprotectant; wherein the cryoprotectant comprises a sugar containing about 6% to about 10% sucrose or trehalose. In some embodiments, the pharmaceutical composition comprising an immune binder, a pharmaceutically acceptable buffer, and a pharmaceutically acceptable stabilizer further comprises a pharmaceutically acceptable cryoprotectant; wherein the cryoprotectant is about 8.0% sucrose.

[0031] In some embodiments, the pharmaceutical composition comprises an immunoconjugate, a pharmaceutically acceptable buffer, and a pharmaceutically acceptable stabilizer; wherein the immunoconjugate comprises (a) a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region containing three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3, and a light chain (LC) variable region containing three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 respectively contain the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, and LC CDR1, LC CDR2, and LC CDR3 respectively contain the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively. (a) the amino acid sequence of 6; (b) an effector bound to the recombinant antibody; wherein the recombinant antibody is bound to the effector, wherein the effector comprises a drug (or a prodrug thereof), a peptide, a protein, a detectable label, a liposome containing a drug (or a prodrug thereof), a radioactive nucleus, a viral particle, or a chelating agent. In some embodiments, the pharmaceutical composition comprises an effector; wherein the effector comprises a drug. In some embodiments, the pharmaceutical composition comprises an effector; wherein the effector comprises an anticancer drug. In some embodiments, the pharmaceutical composition comprises a drug; wherein the drug is a chemotherapeutic agent. In some embodiments, the pharmaceutical composition comprises a drug; wherein the drug is a microtubule inhibitor, a DNA disruptor, or a polymerase inhibitor. In some embodiments, the pharmaceutical composition comprises a drug, namely a microtubule inhibitor; wherein the microtubule inhibitor is aurestatin (or a derivative thereof), sulphurin-10 (or a derivative thereof), or maytansin (or a derivative thereof). In some embodiments, the pharmaceutical composition comprises a drug, namely a microtubule inhibitor; wherein the microtubule inhibitor is monomethylaurestatin F (MMAF), aurestatin E (AE), monomethylaurestatin E (MMAE), valine-citrulline MMAE (vcMMAE), or valine-citrulline MMAF (vcMMAF). In some embodiments, the pharmaceutical composition comprises an immunoconjugate comprising a recombinant antibody and an effector as described above; wherein the ratio of the effector to the recombinant antibody in the group of immunoconjugates is about 3 to about 5. In some embodiments, the pharmaceutical composition comprises an immunoconjugate comprising a recombinant antibody and an effector as described above; wherein the ratio of the effector to the recombinant antibody in the group of immunoconjugates is about 4. In some embodiments, the pharmaceutical composition comprises an immunoconjugate comprising a recombinant antibody and an effector as described above; wherein the effector is bound to the recombinant antibody via a linker.In some embodiments, the pharmaceutical composition comprises an effector that binds to a recombinant antibody via a linker as described above; wherein the linker is a peptide linker, a small molecule linker, or a linker comprising a peptide and a small molecule. In some embodiments, the pharmaceutical composition comprises an effector that binds to a recombinant antibody via a linker as described above; wherein the linker comprises maleiminohexylacetyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB). In another embodiment, the present invention provides a method for treating cancer in a human individual in need, comprising cells expressing CD46, the method comprising administering to the individual an immune conjugate comprising a recombinant antibody, the antibody comprising: a first heavy chain comprising SEQ ID NO: 9, a first light chain comprising SEQ ID NO: 10, a second heavy chain comprising SEQ ID NO: 9, and a second light chain comprising SEQ ID NO: 10; and one, two, three, or four adducts; wherein each of the one, two, three, or four adducts comprises monomethylauratestatin E conjugated to the recombinant antibody via a maleiminohexyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB) linker. (MMAE); wherein each of the first, second, third, or fourth adductor pairs binds to one pair of cysteine ​​residues of the recombinant antibody; wherein the cysteine ​​residue pairs are selected from: C219 of the first heavy chain and C214 of the first light chain; C219 of the second heavy chain and C214 of the second light chain; C225 of the first heavy chain and C225 of the second light chain; and C228 of the first heavy chain and C228 of the second light chain. In some embodiments, a method of treating cancer is provided; wherein the cancer is relapsed or refractory multiple myeloma (RRMM). In some embodiments, a method of treating cancer is provided; wherein the cancer is metastatic castration-resistant prostate cancer (mCRPC). In some embodiments, the method of treating cancer includes administering an immune conjugate to an individual; wherein the immune conjugate comprises two pairs of such adducts. In some embodiments, the method of treating cancer as described above further includes detecting CD46 in the cells. In some embodiments, the method of treating cancer as described above further includes detecting CD46 in the cells; wherein the detection includes immunofluorescence microscopy or immunohistochemistry. In some embodiments, the method of treating cancer as described above further includes detecting CD46 in the cells; wherein the detection includes flow cytometry. In some embodiments, the method of treating cancer as described above further includes detecting CD46 in the cells; wherein the detection includes detecting amplification of chromosome position 1q21. In some embodiments, the method of treating cancer as described above is provided, wherein the immune conjugate is administered to a human individual via intravenous infusion. In some embodiments, the method of treating cancer as described above is provided, wherein the immune conjugate is administered to a human individual every 7 days, every 14 days, every 18 days, every 21 days, or every 30 days. In some embodiments, the method of treating cancer as described above is provided, wherein the immune conjugate is administered to a human individual every 21 days for at least three cycles. In some embodiments, a method of treating cancer as described above is provided, wherein the immune conjugate is administered at a dose of about 1.2 to about 3.0 mg / kg.In some embodiments, a method of treating cancer as described above is provided, wherein the recombinant antibody system is administered at a dose of about 1.8, about 2.4, about 2.7, or about 3.0 mg / kg. In some embodiments, a method of treating cancer as described above is provided, wherein the weight of the desired human individual, in kg, is: the actual weight of the human individual if the actual weight of the human individual is less than the adjusted weight of the individual; the adjusted weight of the human individual if the actual weight of the human individual is greater than or equal to the adjusted weight of the individual and the adjusted weight of the human individual is less than 100 kg; or 100 kg if the adjusted weight of the human individual is greater than or equal to 100 kg. In some embodiments, a method of treating cancer as described above is provided, wherein the weight of the desired human individual, in kg, is the actual weight. In some embodiments, a method of treating cancer as described above is provided, wherein the weight of the desired human individual, in kg, is the adjusted weight.

[0032] In another embodiment, the present invention provides a method for treating metastatic castration-resistant prostate cancer in a human individual in need, the method comprising administering to the individual an immune conjugate comprising: (i) a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region containing three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3, and a light chain comprising a light chain (LC) variable region containing three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 respectively contain the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, and LC CDR1, LC CDR2, and LC CDR3 respectively contain the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6; the antibody is conjugated via a linker to (ii) monomethylauratestatin E. (MMAE) conjugate; wherein the conjugate comprises maleiminohexylidene-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB); wherein the immunoconjugate is administered at a dose of about 1.2 to about 3.0 mg / kg.

[0033] In another embodiment, the present invention provides a method for treating relapsed or refractory multiple myeloma in a human individual in need, the method comprising administering to the individual an immune conjugate comprising: (i) a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region containing three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3, and a light chain comprising a light chain (LC) variable region containing three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 respectively contain the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, and LC CDR1, LC CDR2, and LC CDR3 respectively contain the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6; the antibody is conjugated via a linker to (ii) monomethylauratestatin E. (MMAE) conjugate, wherein the conjugate comprises maleiminohexylacetyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB); wherein the immunoconjugate is administered at a dose of about 1.8 to about 3.0 mg / kg. In some embodiments, a method for treating metastatic castration-resistant prostate cancer in a human individual or a method for treating relapsed or refractory multiple myeloma in a human individual is provided; wherein the calculated weight of the human individual in kg is: the human individual's actual weight if it is less than the individual's adjusted weight; the human individual's adjusted weight if it is greater than or equal to the individual's adjusted weight and the individual's adjusted weight is less than 100 kg; or the human individual's adjusted weight if it is greater than or equal to 100 kg. In some embodiments, a method is provided for treating metastatic castration-resistant prostate cancer or relapsed or refractory multiple myeloma in a human individual; wherein the calculated weight of the human individual, expressed in kg, is an adjusted body weight. In some embodiments, a method is provided for treating metastatic castration-resistant prostate cancer or relapsed or refractory multiple myeloma in a human individual; wherein the weight of the human individual, expressed in kg, is an actual body weight.

[0034] In some embodiments, a method for treating metastatic castration-resistant prostate cancer in a human individual or a method for treating relapsed or refractory multiple myeloma in a human individual further includes detecting CD46 in the cells. In some embodiments, a method for treating metastatic castration-resistant prostate cancer in a human individual or a method for treating relapsed or refractory multiple myeloma in a human individual further includes detecting CD46 in the cells; wherein the detection includes immunofluorescence microscopy or immunohistochemistry. In some embodiments, a method for treating metastatic castration-resistant prostate cancer in a human individual or a method for treating relapsed or refractory multiple myeloma in a human individual further includes detecting CD46 in the cells; wherein the detection includes detecting amplification at chromosome position 1q21. In some embodiments, a method for treating metastatic castration-resistant prostate cancer or relapsed or refractory multiple myeloma in a human individual, as described above, is provided; wherein the immune conjugate is administered to the human individual via intravenous infusion. In some embodiments, a method for treating metastatic castration-resistant prostate cancer or relapsed or refractory multiple myeloma in a human individual, as described above, is provided; wherein the immune conjugate is administered to the human individual every 7 days, every 14 days, every 18 days, every 21 days, or every 30 days. In some embodiments, a method for treating metastatic castration-resistant prostate cancer or relapsed or refractory multiple myeloma in a human individual, as described above, is provided; wherein the immune conjugate is administered to the human individual every 21 days for at least three cycles.

[0035] In another embodiment, the present invention provides a method for treating cancer in a human individual in need, the method comprising administering to the human individual an immune conjugate comprising: a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region including three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3; and a light chain (LC) variable region including three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 respectively comprise the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, and LC CDR1, LC CDR2, and LC CDR3 respectively comprise the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6; an effector binding to the recombinant antibody; and wherein the immunomodulator is administered at a dose of about 1.0 to about 5.0 mg / kg or at a dose of about 1.0 to about 4.0 mg / kg. A dose of mg / kg is administered. In some embodiments, a method of treating cancer is provided; wherein the cancer is prostate cancer. In some embodiments, a method of treating prostate cancer is provided; wherein the prostate cancer is metastatic castration-resistant prostate cancer. In some embodiments, a method of treating cancer is provided; wherein the cancer is multiple myeloma. In some embodiments, a method of treating multiple myeloma is provided; wherein the multiple myeloma is relapsed or refractory multiple myeloma. In some embodiments, the method of treating cancer as described above further includes detecting CD46 expression in the cells of the cancer. In some embodiments, the method of treating cancer as described above further includes detecting CD46 expression in the cells of the cancer, wherein the detection includes immunofluorescence microscopy or immunohistochemistry. In some embodiments, the method of treating cancer as described above further includes detecting CD46 expression in the cells of the cancer, wherein the detection includes flow cytometry. In some embodiments, the method of treating cancer as described above further includes detecting CD46 expression in the cells of the cancer, wherein the detection includes detecting amplification at chromosome position 1q21. In some embodiments, a method for treating cancer as described above is provided; wherein the cancer has higher CD46 expression than non-cancerous tissue of the same tissue type from the individual or from a healthy individual. In some embodiments, a method for treating cancer as described above is provided; wherein the cancer comprises an increase in the copy number of chromosome 1q21.In some embodiments, a method of treating cancer as described above is provided; wherein the immune conjugate is administered at concentrations of approximately 1.0 to approximately 4.5 mg / kg, approximately 1.0 to approximately 4.0 mg / kg, approximately 1.0 to approximately 3.5 mg / kg, approximately 1.0 to approximately 3.0 mg / kg, approximately 1.0 to approximately 2.7 mg / kg, approximately 1.0 to approximately 2.5 mg / kg, approximately 1.0 to approximately 2.4 mg / kg, approximately 1.5 to approximately 4.5 mg / kg, approximately 1.5 to approximately 4.0 mg / kg, approximately 1.5 to approximately 3.5 mg / kg, approximately 1.5 to approximately 3.0 mg / kg, approximately 1.5 to approximately 2.7 mg / kg, approximately 1.5 to approximately 2.5 mg / kg, approximately 1.5 to approximately 2.5 mg / kg, approximately 1.5 to approximately 2.4 mg / kg, approximately 1.5 to approximately 2.0 mg / kg, approximately 1.8 to approximately 4.5 mg / kg, approximately 1.8 to approximately 4.0 mg / kg, approximately 1.8 to approximately 3.5 mg / kg, and approximately 1.8 to approximately 3.0 mg / kg. The immune conjugate is administered at a dose of about 1.8 to about 2.7 mg / kg, about 1.8 to about 2.5 mg / kg, about 1.8 to about 2.4 mg / kg, or about 1.8 to about 2.0 mg / kg. In some embodiments, a method of treating cancer as described above is provided; wherein the immune conjugate is administered at a dose of about 1.2 to about 3.0 mg / kg. In some embodiments, a method of treating cancer as described above is provided; wherein the immune conjugate is administered at a dose of about 1.0, about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2.0, about 2.1, about 2.2, about 2.3, about 2.4, about 2.5, about 2.6, about 2.7, about 2.8, about 2.9, about 3.0, about 3.1, about 3.2, about 3.3, about 3.4, about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, or about 4.0 mg / kg. In some embodiments, a method of treating cancer as described above is provided; wherein the immune conjugate is administered at a dose of about 1.8, about 2.4, about 2.7, or about 3.0 mg / kg. In some embodiments, a method of treating cancer as described above is provided; wherein the immune conjugate is administered at a dose of about 1.8 mg / kg. In some embodiments, a method of treating cancer as described above is provided; wherein the immune conjugate is administered at a dose of about 2.4 mg / kg. In some embodiments, a method of treating cancer as described above is provided; wherein the immune conjugate is administered at a dose of about 2.7 mg / kg. In some embodiments, a method of treating cancer as described above is provided; wherein the immune conjugate is administered at a dose of about 3.0 mg / kg.In some embodiments, a method for treating cancer in a human individual as described above is provided; wherein the weight of the human individual, in kg, is: the actual weight of the human individual if the actual weight of the human individual is less than the adjusted weight of the individual; the adjusted weight of the human individual if the actual weight of the human individual is greater than or equal to the adjusted weight of the individual and the adjusted weight of the human individual is less than 100 kg; or 100 kg if the adjusted weight of the human individual is greater than or equal to 100 kg. In some embodiments, a method for treating cancer in a human individual as described above is provided; wherein the weight of the human individual, in kg, is the actual weight. In some embodiments, a method for treating cancer in a human individual as described above is provided; wherein the weight of the human individual, in kg, is the adjusted weight. In some embodiments, a method for treating cancer in a human individual is provided, the method comprising administering to the human individual an immune conjugate comprising a recombinant antibody as described above; wherein the recombinant antibody system is administered to the human individual via intravenous infusion. In some embodiments, a method of treating cancer in a human individual is provided, the method comprising administering to the human individual an immune conjugate containing a recombinant antibody as described above; wherein the recombinant antibody system is administered to the human individual every 7 days, every 14 days, every 18 days, every 21 days, or every 30 days. In some embodiments, a method of treating cancer in a human individual is provided, the method comprising administering to the human individual an immune conjugate containing a recombinant antibody as described above; wherein the recombinant antibody system is administered to the human individual every 21 days for at least three cycles. In some embodiments, a method of treating cancer in a human individual is provided, the method comprising administering to the human individual an immune conjugate containing an effector as described above; wherein the effector comprises a drug (or a prodrug thereof), a peptide, a protein, a detectable marker, a liposome containing a drug (or a prodrug thereof), a radionuclides, viral particles, or a chelating agent. In some embodiments, a method of treating cancer in a human individual is provided, the method comprising administering to the human individual an immune conjugate containing an effector as described above; wherein the effector comprises a drug. In some embodiments, a method for treating cancer in a human individual as described above is provided; wherein the effector comprises an anticancer drug. In some embodiments, a method for treating cancer in a human individual as described above is provided; wherein the effector comprises a drug; wherein the drug is a chemotherapeutic agent. In some embodiments, a method for treating cancer in a human individual as described above is provided; wherein the effector comprises a drug; wherein the drug is a microtubule inhibitor, a DNA disruptor, or a polymerase inhibitor. In some embodiments, a method for treating cancer in a human individual as described above is provided; wherein the effector comprises a drug; wherein the drug is a microtubule inhibitor.In some embodiments, a method for treating cancer in a human individual as described above is provided; wherein the microtubule inhibitor is aurestatin (or a derivative thereof), sulphurin-10 (or a derivative thereof), or maytansin (or a derivative thereof). In some embodiments, a method for treating cancer in a human individual as described above is provided; wherein the microtubule inhibitor is monomethylaurestatin F (MMAF), aurestatin E (AE), monomethylaurestatin E (MMAE), valine-citrulline MMAE (vcMMAE), or valine-citrulline MMAF (vcMMAF). In some embodiments, a method for treating cancer in a human individual as described above is provided; wherein the microtubule inhibitor is monomethylaurestatin E (MMAE). In some embodiments, a method for treating cancer in a human individual is provided, the method comprising administering to the human individual an immune conjugate comprising an effector and a recombinant antibody as described above; wherein the ratio of the effector to the recombinant antibody is about 3 to about 5. In some embodiments, a method for treating cancer in a human individual as described above is provided; wherein the ratio of the effector to the recombinant antibody is approximately 4. In some embodiments, a method for treating cancer in a human individual as described above is provided; wherein the effector is bound to the recombinant antibody via a linker. In some embodiments, a method for treating cancer in a human individual as described above is provided; wherein the linker is a peptide linker, a small molecule linker, or a linker comprising a peptide and a small molecule. In some embodiments, a method for treating cancer in a human individual as described above is provided; wherein the linker comprises maleiminohexyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB). In some embodiments, a method for treating cancer in a human individual as described above is provided; wherein the immune conjugate binds to CD46 expressed on the cell surface and is internalized into the cell. In some embodiments, a method for treating cancer in a human individual as described above is provided; wherein the immune conjugate is internalized into the cell via macropinocytosis.

[0036] In another embodiment, the present invention provides an immune conjugate comprising: a recombinant antibody comprising: a first heavy chain of SEQ ID NO: 9, a first light chain of SEQ ID NO: 10, a second heavy chain of SEQ ID NO: 9, and a second light chain of SEQ ID NO: 10; and one, two, three, or four adducts; wherein each of the one, two, three, or four adducts comprises monomethylauratestatin E conjugated to the recombinant antibody via a maleiminohexyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB) linker. (MMAE); wherein each of the first, second, third, or fourth adductor pairs binds to one pair of cysteine ​​residues of the recombinant antibody, wherein the cysteine ​​residue pairs are selected from: C219 of the first heavy chain and C214 of the first light chain, C219 of the second heavy chain and C214 of the second light chain, C225 of the first heavy chain and C225 of the second light chain, and C228 of the first heavy chain and C228 of the second light chain; for the treatment of cancers in human individuals containing cells expressing CD46. In some embodiments, an immune conjugate as described above is provided for the treatment of cancers in human individuals, wherein the cancer is relapsed or refractory multiple myeloma (RRMM). In some embodiments, an immune conjugate as described above is provided for the treatment of cancers in human individuals, wherein the cancer is metastatic castration-resistant prostate cancer (mCRPC). In some embodiments, an immune conjugate as described above is provided for the treatment of cancers in human individuals, wherein the immune conjugate comprises two pairs of such adductor pairs. In some embodiments, an immune conjugate as described above is provided for treating cancer in a human individual comprising cells expressing CD46, wherein the cells comprise CD46 as determined by immunofluorescence microscopy or immunohistochemistry. In some embodiments, an immune conjugate as described above is provided for treating cancer in a human individual comprising cells expressing CD46, wherein the cells comprise CD46 as determined by flow cytometry. In some embodiments, an immune conjugate as described above is provided for treating cancer in a human individual comprising cells expressing CD46, wherein the cells comprise an amplification of chromosome position 1q21. In some embodiments, an immune conjugate as described above is provided for treating cancer in a human individual, wherein the immune conjugate is formulated for intravenous infusion. In some embodiments, an immune conjugate as described above is provided for treating cancer in a human individual, wherein the immune conjugate is administered to the human individual every 7 days, every 14 days, every 18 days, every 21 days, every 28 days, or monthly. In some embodiments, an immune conjugate as described above for treating cancer in a human individual is provided, wherein the immune conjugate is administered to the human individual every 21 days.In some embodiments, an immune conjugate as described above for treating cancer in a human individual is provided, wherein the immune conjugate is administered at a dose of about 1.2 to about 3.0 mg / kg. In some embodiments, an immune conjugate comprising a recombinant antibody as described above for treating cancer in a human individual is provided, wherein the recombinant antibody is administered at a dose of about 1.8, about 2.4, about 2.7, or about 3.0 mg / kg. In some embodiments, an immune conjugate as described above for treating cancer in a human individual is provided; wherein the weight of the human individual, in kg, is: the human individual's actual weight if the human individual's actual weight is less than the individual's adjusted weight; the human individual's adjusted weight if the human individual's actual weight is greater than or equal to the individual's adjusted weight and the human individual's adjusted weight is less than 100 kg; or 100 kg if the human individual's adjusted weight is greater than or equal to 100 kg. In some embodiments, an immune conjugate as described above is provided for treating cancer in a human individual; wherein the weight of the human individual, expressed in kg, is the actual body weight. In some embodiments, an immune conjugate as described above is provided for treating cancer in a human individual; wherein the weight of the human individual, expressed in kg, is the adjusted body weight.

[0037] In another embodiment, the present invention provides an immune conjugate for treating metastatic castration-resistant prostate cancer in human individuals of need, comprising (i) a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region containing three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3, and a light chain containing a light chain (LC) variable region containing three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 contain the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and LC CDR1, LC CDR2, and LC CDR3 contain the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively; the antibody is conjugated via a linker to (ii) monomethylauratestatin E. (MMAE) conjugate, wherein the conjugate comprises maleiminohexylidene-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB), wherein the immunoconjugate is administered at a dose of about 1.2 to about 3.0 mg / kg.

[0038] In another embodiment, the present invention provides an immune conjugate for treating refractory multiple myeloma in human individuals in need, comprising (i) a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region containing three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3, and a light chain containing a light chain (LC) variable region containing three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 contain the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and LC CDR1, LC CDR2, and LC CDR3 contain the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively; the antibody is conjugated via a linker to (ii) monomethylauratestatin E. (MMAE) conjugate, wherein the conjugate comprises maleiminohexylidene-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB), wherein the immunoconjugate is administered at a dose of about 1.8 to about 3.0 mg / kg.

[0039] In some embodiments, an immune conjugate is provided for treating metastatic castration-resistant prostate cancer or refractory multiple myeloma in a human individual, wherein the calculated weight of the human individual, in kg, is: the human individual's actual weight if the human individual's actual weight is less than the individual's adjusted weight; the human individual's adjusted weight if the human individual's actual weight is greater than or equal to the individual's adjusted weight and the individual's adjusted weight is less than 100 kg; or 100 kg if the human individual's adjusted weight is greater than or equal to 100 kg. In some embodiments, an immune conjugate is provided for treating metastatic castration-resistant prostate cancer or refractory multiple myeloma in a human individual, wherein the weight of the human individual, in kg, is the actual weight. In some embodiments, an immune conjugate is provided for treating metastatic castration-resistant prostate cancer or refractory multiple myeloma in a human individual, wherein the weight of the human individual, in kg, is the adjusted weight. In some embodiments, an immune conjugate is provided for treating metastatic castration-resistant prostate cancer or refractory multiple myeloma in a human individual, wherein the cancer comprises a cell containing CD46 as determined by immunofluorescence microscopy or immunohistochemistry. In some embodiments, an immune conjugate is provided for treating metastatic castration-resistant prostate cancer or refractory multiple myeloma in a human individual, wherein the cancer comprises a cell containing CD46 as determined by flow cytometry. In some embodiments, an immune conjugate is provided for treating metastatic castration-resistant prostate cancer or refractory multiple myeloma in a human individual, wherein the cancer comprises a cell containing an amplification of chromosome position 1q21. In some embodiments, an immune conjugate is provided for treating metastatic castration-resistant prostate cancer or refractory multiple myeloma in a human individual; wherein the immune conjugate is formulated for intravenous infusion. In some embodiments, an immune conjugate is provided for treating metastatic castration-resistant prostate cancer or refractory multiple myeloma in a human individual; wherein the immune conjugate is administered to the human individual every 7 days, every 14 days, every 18 days, every 21 days, every 28 days, or monthly. In some embodiments, an immune conjugate is provided for treating metastatic castration-resistant prostate cancer or refractory multiple myeloma in a human individual; wherein the immune conjugate is administered to the human individual every 21 days for at least three cycles.

[0040] In another embodiment, the present invention provides an immune conjugate for treating cancer in human individuals in need, the immune conjugate comprising: (a) a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region comprising three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3, and a light chain (LC) variable region comprising three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 comprise the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and LC CDR1, LC CDR2, and LC CDR3 comprise the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively; (b) an effector that binds to the recombinant antibody; and wherein the immune conjugate is administered at a dose of about 1.0 to about 5.0 mg / kg. In some embodiments, an immune conjugate is provided for treating cancer in a human individual, wherein the cancer is prostate cancer. In some embodiments, an immune conjugate is provided for treating cancer in a human individual, wherein the prostate cancer is metastatic castration-resistant prostate cancer. In some embodiments, an immune conjugate is provided for treating cancer in a human individual, wherein the cancer is multiple myeloma. In some embodiments, an immune conjugate is provided for treating multiple myeloma in a human individual, wherein the multiple myeloma is relapsed or refractory multiple myeloma. In some embodiments, an immune conjugate is provided for treating cancer in a human individual, wherein the cancer comprises cells expressing CD46 as determined by immunofluorescence microscopy or immunohistochemistry. In some embodiments, an immune conjugate is provided for treating cancer in a human individual, wherein the cancer comprises cells expressing CD46 as determined by flow cytometry. In some embodiments, an immune conjugate is provided for treating cancer in a human individual, wherein the cancer comprises amplification of chromosome position 1q21. In some embodiments, an immune conjugate is provided for treating cancer in a human individual; wherein the cancer has higher CD46 expression than non-cancerous tissue of the same tissue type from the individual or from a healthy individual.In some embodiments, an immune conjugate is provided for treating cancer in a human individual; wherein the immune conjugate is prepared at a concentration of about 1.0 to about 4.5 mg / kg, about 1.0 to about 4.0 mg / kg, about 1.0 to about 3.5 mg / kg, about 1.0 to about 3.0 mg / kg, about 1.0 to about 2.7 mg / kg, about 1.0 to about 2.5 mg / kg, about 1.0 to about 2.4 mg / kg, about 1.5 to about 4.5 mg / kg, about 1.5 to about 4.0 mg / kg, about 1.5 to about 3.5 mg / kg, about 1.5 to about 3.0 mg / kg, about 1.5 to about 2.7 mg / kg, about 1.5 to about 2.5 mg / kg, about 1.5 to about 2.4 mg / kg, about 1.5 to about 2.0 mg / kg, about 1.8 to about 4.5 mg / kg, about 1.8 to about 4.0 mg / kg, about 1.8 to about 3.5 mg / kg, or about 1.8 to about 3.0 mg / kg. The drug is administered at doses of approximately 1.8 to approximately 2.7 mg / kg, approximately 1.8 to approximately 2.5 mg / kg, approximately 1.8 to approximately 2.4 mg / kg, or approximately 1.8 to approximately 2.0 mg / kg. In some embodiments, an immune conjugate is provided for treating cancer in a human individual; wherein the immune conjugate is administered at a dose of approximately 1.2 to approximately 3.0 mg / kg. In some embodiments, an immune conjugate is provided for treating cancer in a human individual; wherein the immune conjugate is administered at a dose of about 1.0, about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2.0, about 2.1, about 2.2, about 2.3, about 2.4, about 2.5, about 2.6, about 2.7, about 2.8, about 2.9, about 3.0, about 3.1, about 3.2, about 3.3, about 3.4, about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, or about 4.0 mg / kg. In some embodiments, an immune conjugate is provided for treating cancer in a human individual; wherein the immune conjugate is administered at a dose of about 1.8, about 2.4, about 2.7, or about 3.0 mg / kg. In some embodiments, an immune conjugate for treating cancer in a human individual is provided; wherein the immune conjugate is administered at a dose of about 1.8, about 2.4, about 2.7, or about 3.0 mg / kg. In some embodiments, an immune conjugate for treating cancer in a human individual is provided; wherein the immune conjugate is administered at a dose of about 1.8 mg / kg. In some embodiments, an immune conjugate for treating cancer in a human individual is provided; wherein the immune conjugate is administered at a dose of about 2.4 mg / kg. In some embodiments, an immune conjugate for treating cancer in a human individual is provided; wherein the immune conjugate is administered at a dose of about 2.7 mg / kg.In some embodiments, an immune conjugate for treating cancer in a human individual is provided; wherein the immune conjugate is administered at a dose of about 3.0 mg / kg. In some embodiments, an immune conjugate for treating cancer in a human individual is provided; wherein the weight of the human individual in kg is: the actual weight of the human individual if the actual weight of the human individual is less than the adjusted weight of the individual; the adjusted weight of the human individual if the actual weight of the human individual is greater than or equal to the adjusted weight of the individual and the adjusted weight of the human individual is less than 100 kg; or 100 kg if the adjusted weight of the human individual is greater than or equal to 100 kg. In some embodiments, an immune conjugate for treating cancer in a human individual is provided; wherein the weight of the human individual in kg is the actual weight. In some embodiments, an immune conjugate for treating cancer in a human individual is provided; wherein the weight of the human individual in kg is the adjusted weight. In some embodiments, an immune conjugate comprising a recombinant antibody for treating cancer in a human individual is provided; wherein the recombinant antibody is formulated for intravenous infusion. In some embodiments, an immune conjugate comprising a recombinant antibody is provided for treating cancer in a human individual; wherein the recombinant antibody system is administered to the human individual every 7 days, every 14 days, every 18 days, every 21 days, or every 30 days. In some embodiments, an immune conjugate comprising a recombinant antibody is provided for treating cancer in a human individual; wherein the recombinant antibody system is administered to the human individual every 21 days. In some embodiments, an immune conjugate comprising an effector is provided for treating cancer in a human individual; wherein the effector comprises a drug (or a prodrug thereof), a peptide, a protein, a detectable label, a liposome containing a drug (or a prodrug thereof), a radioactive nucleus, a viral particle, or a chelating agent. In some embodiments, an immune conjugate comprising an effector is provided for treating cancer in a human individual; wherein the effector comprises a drug. In some embodiments, an immune conjugate comprising an effector is provided for treating cancer in a human individual; wherein the effector comprises an anticancer drug. In some embodiments, an immune conjugate comprising an effector is provided; wherein the effector comprises a chemotherapeutic agent. In some embodiments, an immune conjugate comprising an effector is provided; wherein the effector comprises a drug; wherein the drug is a microtubule inhibitor, a DNA disruptor, or a polymerase inhibitor. In some embodiments, an immune conjugate comprising a microtubule inhibitor is provided, wherein the microtubule inhibitor is aurestatin (or a derivative thereof), sulphurin-10 (or a derivative thereof), or maytansin (or a derivative thereof). In some embodiments, an immune conjugate comprising a microtubule inhibitor is provided, wherein the microtubule inhibitor is monomethylaurestatin F (MMAF), aurestatin E (AE), monomethylaurestatin E (MMAE), valine-citrulline MMAE (vcMMAE), or valine-citrulline MMAF (vcMMAF).In some embodiments, an immune conjugate comprising an effector is provided; wherein the effector comprises a drug; wherein the drug is a microtubule inhibitor, and wherein the microtubule inhibitor is monomethylaurestatin E (MMAE). In some embodiments, an immune conjugate comprising an effector and a recombinant antibody is provided; wherein the ratio of the effector to the recombinant antibody is about 3 to about 5. In some embodiments, an immune conjugate comprising an effector and a recombinant antibody is provided; wherein the ratio of the effector to the recombinant antibody is about 4. In some embodiments, an immune conjugate comprising an effector and a recombinant antibody is provided; wherein the effector is bound to the recombinant antibody via a linker. In some embodiments, the immune conjugate comprises an effector bound to the recombinant antibody via a linker; wherein the linker is a peptide linker, a small molecule linker, or a linker comprising a peptide and a small molecule. In some embodiments, the immune conjugate comprises an effector bound to the recombinant antibody via a linker; wherein the linker comprises maleiminohexyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB). In some embodiments, an immunoconjugate comprising an effector and a recombinant antibody is provided; wherein the immunoconjugate binds to CD46 expressed on the surface of a cell and is internalized into the cell. In some embodiments, an immunoconjugate comprising an effector and a recombinant antibody is provided; wherein the immunoconjugate is internalized into the cell via macropinocytosis.

[0041] In another embodiment, the present invention provides a pharmaceutical formulation for treating metastatic castration-resistant prostate cancer in human individuals in need, comprising an immunoconjugate at a concentration of about 10.0 ± 1.0 mg / mL, about 20 mM histidine buffer, about 8.0% sucrose, and about 0.01% polysorbate 80; wherein the immunoconjugate comprises: a recombinant antibody comprising: a first heavy chain of SEQ ID NO: 9, a first light chain of SEQ ID NO: 10, a second heavy chain of SEQ ID NO: 9, and a second light chain of SEQ ID NO: 10; and one, two, three, or four adducts; wherein each of the one, two, three, or four adducts comprises monomethylauratestatin E conjugated to the recombinant antibody via a maleiminohexyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB) linker. (MMAE); wherein each of the first, second, third, or fourth adduct pairs binds to one pair of cysteine ​​residues of the recombinant antibody, wherein such cysteine ​​residue pairs are selected from: C219 of the first heavy chain and C214 of the first light chain; C219 of the second heavy chain and C214 of the second light chain; C225 of the first heavy chain and C225 of the second light chain; and C228 of the first heavy chain and C228 of the second light chain.

[0042] In another embodiment, the present invention provides a pharmaceutical formulation for treating refractory multiple myeloma in human individuals in need, comprising an immunoconjugate at a concentration of about 10.0 ± 1.0 mg / mL, about 20 mM histidine buffer, about 8.0% sucrose, and about 0.01% polysorbate 80; wherein the immunoconjugate comprises: a recombinant antibody comprising: a first heavy chain of SEQ ID NO: 9, a first light chain of SEQ ID NO: 10, a second heavy chain of SEQ ID NO: 9, and a second light chain of SEQ ID NO: 10; and one, two, three, or four adducts; wherein each of the one, two, three, or four adducts comprises monomethylauratestatin E conjugated to the recombinant antibody via a maleiminohexyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB) linker. (MMAE); wherein each of the first, second, third, or fourth adduct pairs binds to one pair of cysteine ​​residues of the recombinant antibody, wherein such cysteine ​​residue pairs are selected from: C219 of the first heavy chain and C214 of the first light chain; C219 of the second heavy chain and C214 of the second light chain; C225 of the first heavy chain and C225 of the second light chain; and C228 of the first heavy chain and C228 of the second light chain. Simple Explanation of the Diagram

[0043] [picture] [1] The flow cytometry trajectory of YS5FL bound to prostate cells is shown.

[0044] [picture] [2] The flow cytometry trajectory of YS5FL bound to multiple myeloma cells is illustrated.

[0045] [picture] [3] is a schematic diagram showing the structure of the FOR46 immune conjugate described herein.

[0046] [picture] [4] shows the stoichiometric hydrophobic interaction trajectory of the binding of MMAE and YS5FL in FOR46.

[0047] [picture] [5A] is a CT scan showing metastatic lesions in a castration-resistant prostate cancer patient 001-09-28 (administered with 2.7 mg / kg FOR46) on day 15 of cycle 3 and before treatment.

[0048] [picture] [5B] is a diagram illustrating the decrease in serum PSA in the patient on 001-09-28.

[0049] [picture] [6] is a swimming graph showing the status of patients in a dose escalation trial for prostate cancer. PR: Partial response; EOS: End of study; EOT: End of treatment; PD: Progressive disease.

[0050] [picture] [7A] is a diagram showing the response of a multiple myeloma patient 006-05-008 to treatment with 1.8 mg / kg FOR46.

[0051] [picture] [7B] is a diagram showing the response of a multiple myeloma patient 001-06-012 to treatment with 2.4 mg / kg FOR46.

[0052] [picture] [7C] is a diagram showing the response of a multiple myeloma patient 003-06-014 to treatment with 2.4 mg / kg FOR46.

[0053] [picture] [8] Swimming graph showing the status of patients in the dose escalation and expansion trials of multiple myeloma. EOS: End of study; EOT: End of treatment; PD: Progressive disease. Implementation

[0054] [Cross-reference]

[0055] This application claims the benefit of U.S. Provisional Application No. 63 / 062,740, filed August 7, 2020, which is incorporated herein by reference in its entirety.

[0056] CD46, also known as CD46 complement regulatory protein, differentiation cluster 46, and membrane cofactor protein, is an inhibitory complement receptor. Overexpression of CD46 has been observed in several cancers, such as breast cancer, colorectal cancer, liver cancer, lung cancer, and prostate cancer. In some cases, CD46 overexpression is a characteristic of negative prognostic factors. For example, CD46 overexpression has been associated with shorter progression-free survival and shorter overall survival in patients with breast and ovarian cancer. This article provides CD46-targeting antibodies and immune conjugates for the treatment of cancer. This article further provides specific dosing and administration regimens for delivering CD46-targeting antibodies and immune conjugates to individuals in need. This article further provides formulations of CD46-targeting antibodies and immune conjugates for administration to individuals in need, providing, for example, adequate stability and cryoprotection. [definition]

[0057] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one skilled in the art to which the claimed subject matter pertains. It should be understood that the foregoing general description and the following detailed description are merely illustrative and explanatory and do not limit any of the claimed subject matter. Section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter. In this application, the singular is used to include the plural unless otherwise specifically stated. It should be noted that, unless the context clearly requires otherwise, the singular forms “a / an” and “the” as used in the specification and the appended claims include plural indicators. In this application, the use of “or” means “and / or” unless otherwise stated. Furthermore, the use of the term “including” and other forms such as “include / includes / included” is not restrictive.

[0058] As used herein, ranges and quantities can be expressed as “about” a specific value or range. “About” also includes exact quantities. Therefore, “about 5 µL” means both “about 5 µL” and “5 µL”. Generally, the term “about” includes quantities expected to be within experimental error.

[0059] The terms “antibody” and “immunoglobulin” are used interchangeably in this document and are used in the broadest sense to encompass fully assembled antibodies, antibody fragments that can bind to antigens (e.g., Fab, F(ab')2, Fv, single-chain antibodies (scFv)), diabodies, antibody chimeras, hybrid antibodies, bispecific antibodies, and similar antibodies.

[0060] The terms "monoclonal antibody" and "mAb" are used interchangeably in this document and refer to antibodies derived from substantially homogeneous antibody groups, that is, individual antibody systems of the antibody group are identical except for the possible small number of naturally occurring mutations.

[0061] The terms "natural antibody" and "natural immunoglobulin" refer to heterotetrameric glycoproteins of approximately 150,000 Daltons, composed of two identical light (L) chains and two identical heavy (H) chains. Each light chain is connected to the heavy chain by a covalent disulfide bond, and the number of disulfide bonds varies in the heavy chains of different immunoglobulin isoforms. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has a variable domain (VH) followed by multiple constant domains at one end. Each light chain has a variable domain (VL) at one end and a constant domain at the other end; the constant domains of the light chain are aligned with the first constant domain of the heavy chain, and the variable domains of the light chain are aligned with the variable domains of the heavy chain. It is believed that specific amino acid residues form the interface between the variable domains of the light and heavy chains.

[0062] As used in this article, the term "highly variable region" refers to the amino acid residues of the antibody responsible for binding to the antigen. The highly variable region includes amino acid residues from the complementarity-determining region or the CDR (i.e., residues 24-34 (L1), 50-56 (L2), and 89-97 (L3) in the light chain variable domain and residues 31-35 (H1), 50-65 (H2), and 95-102 (H3) in the heavy chain variable domain; Kabat et al. (1991) Sequences of Proteins of Immunological Interest, 5th Edition, US Department of Health and Human Services, NIH Publication No. 91-3242 (referred to as "Kabat et al." in this paper) and / or residues from the highly variable ring (i.e., residues 26-32 (L1), 50-52 (L2), and 91-96 (L3) in the light chain variable domain and residues (H1), 53-55 (H2), and 96-101 (13) in the heavy chain variable domain); Chothia and Lesk, (1987) J. Mol. Biol., 196:901-917). The “framework” or “FR” residues are those variable domain residues other than the highly variable region residues as believed in this article.

[0063] In some cases, the CDR of an antibody is determined according to: (i) the Kabat numbering system, Kabat et al. (1991) Sequences of Proteins of Immunological Interest, 5th edition, US Department of Health and Human Services, NIH Publication No. 91-3242; or (ii) the Chothia numbering system, which will be referred to herein as “Chothia CDR” (see, for example, Chothia and Lesk, 1987, J. Mol. Biol., 196:901-917; Al-Lazikani et al., 1997, J. Mol. Biol., 273:927-948; Chothia et al., 1992, J. Mol. Biol., 227:799-817; Tramontano A et al., 1990, J. Mol. Biol. 215(1): 175-82; and U.S. Patent No. 7,709,226); or (iii) the ImMunoGeneTics (IMGT) numbering system, as described, for example, in Lefranc, M.-P., 1999, The Immunologist, 7: 132-136 and Lefranc, M.-P. et al., 1999, Nucleic Acids Res., 27:209-212 (“IMGT CDR”); or (iv) MacCallum et al., 1996, J. Mol. Biol., 262:732-745. See also, for example, Martin, A., “Protein Sequence and Structure Analysis of Antibody Variable Domains,” in Antibody Engineering, Kontermann and Diibel eds., Chapter 31, pp. 422-439, Springer-Verlag, Berlin (2001).

[0064] Regarding the Kabat numbering system, the CDR in antibody heavy chain molecules is typically located at amino acid positions 31 to 35, and may, depending on the circumstances, include one or two other amino acids following amino acid positions 35 (referred to as 35A and 35B in the Kabat numbering system) (CDR1), 50 to 65 (CDR2), and 95 to 102 (CDR3). Using the Kabat numbering system, the CDR in antibody light chain molecules is typically located at amino acid positions 24 to 34 (CDR1), 50 to 56 (CDR2), and 89 to 97 (CDR3). As is well known to those skilled in the art, using the Kabat numbering system, the actual linear amino acid sequence of the antibody variable domain may contain fewer or other amino acids due to shortening or lengthening of the FR and / or CDR, and therefore, the Kabat number of an amino acid may not be the same as its linear amino acid number.

[0065] As used herein, the term "antigen binding site" refers to the portion of an antigen-binding molecule that binds specifically to an antigenic determinant. More specifically, the term "antigen binding site" refers to a portion of an antibody that includes a region that binds specifically to a portion or the whole of an antigen and is complementary to that portion or the whole of the antigen. When the antigen is large, the antigen-binding molecule may bind only to a specific portion of the antigen, which is called an antigenic determinant. Antigen binding sites may be provided by, for example, one or more variable domains (also called variable regions). Preferably, the antigen binding site includes a variable region (VL) of the antibody light chain and a variable region (VH) of the antibody heavy chain.

[0066] "Specific binding" means that the binding is selective for the antigen and can be distinguished from unwanted or non-specific interactions. The ability of an antigen-binding molecule to bind to a specific antigen can be measured by enzyme-linked immunosorbent assay (ELISA) or other techniques familiar to those skilled in this technique (e.g., surface plasma resonance (SPR) technique (analyzed on a BIAcore instrument) (Liljeblad et al., Glyco J 17, 323-329 (2000)) and conventional binding assays (Heeley, Endocr Res 28, 217-229 (2002)). In one embodiment, as measured, for example by SPR, the degree to which an antigen-binding molecule binds to an unrelated protein is less than about 10% of the degree to which the antigen-binding molecule binds to the antigen. In a particular embodiment, the molecule that binds to the antigen has a dissociation constant (Kd) of ≤1 μM, ≤100 nM, ≤10 nM, ≤1 nM, ≤0.1 nM, ≤0.01 nM or ≤0.001 nM (e.g. 10⁻⁷ M or less, e.g. 10⁻⁷ M to 10⁻¹³ M, e.g. 10⁻⁹ M to 10⁻¹³ M).

[0067] Immunoglobulins can be classified into different classes based on the amino acid sequence of their heavy chain constant domains. There are five main classes of human immunoglobulins: IgA, IgD, IgE, IgG, IgM, and IgY, and some of these can be further subdivided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known. Different isotypes have different effector functions. For example, human IgG1 and IgG3 isotypes have ADCC (antibody-dependent cell-mediated cytotoxicity) activity. The light chains of antibodies (immunoglobulins) from any vertebrate species can be aligned with one of two distinct types (called κ and λ) based on the amino acid sequence of their constant domains.

[0068] As used herein, "chimeric antibody" refers to an antibody in which a portion of the heavy chain and / or light chain is derived from a specific source (e.g., protein) or species, while the remaining portion of the heavy chain and / or light chain is derived from a different source (e.g., protein) or species.

[0069] As used herein, the term "recombinant human antibody" is intended to include all human antibodies prepared, expressed, manufactured, or isolated by recombinant means, such as antibodies isolated from host cells such as NSO or CHO cells, antibodies isolated from transgenic animals (e.g., mice) targeting human immunoglobulin genes, or antibodies expressed using a recombinant expression vector transfected into host cells. Such recombinant human antibodies have variable and constant regions arranged in a rearranged manner. In some cases, recombinant human antibodies have undergone in vivo somatic hypermutation. Therefore, the amino acid sequences of the VH and VL regions of recombinant antibodies are derived from and associated with human germline VH and VL sequences, but may not be sequences naturally present in the human antibody germline lineage in vivo.

[0070] As used herein, the term "valence" indicates the presence of a specified number of binding sites in an antigen-binding molecule. Therefore, the terms "bivalent," "tetravalent," and "hexavalent" respectively indicate the presence of two, four, and six binding sites in an antigen-binding molecule. The bispecific antibody system according to the invention is at least "bivalent" and may be "trivalent" or "multivalent" (e.g., "tetravalent" or "hexavalent"). In a specific state, the antibody of the invention has two or more binding sites and is bispecific. That is, even in the presence of more than two binding sites (i.e., a trivalent or multivalent antibody system), the antibody may be bispecific. Specifically, the invention relates to a bispecific bivalent antibody having a binding site for each antigen that binds to it in a specific manner.

[0071] As used herein, a "monospecific" antibody means an antibody having one or more binding sites, each of which binds to the same antigenic determinant of the same antigen.

[0072] The terms "individual / subject" and "patient" are used interchangeably herein and refer to any mammal. In some embodiments, the mammal is a human. In some embodiments, the mammal is a non-human. Neither of the terms requires, nor is limited to, supervision (e.g., continuous or intermittent supervision) by a healthcare worker (e.g., physician, registered nurse, nurse practitioner, physician assistant, nursing home or hospice worker).

[0073] As used herein, the term "amino acid sequence identity percentage (%)" is defined as, if necessary, the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in a given sequence after alignment and the introduction of gaps to achieve the maximum sequence identity percentage, without considering any retained substitutions as part of the sequence identity. Alignment for the purpose of determining the amino acid sequence identity percentage can be achieved in various ways within the art, for example using publicly available computer software such as EMBOSS MATCHER, EMBOSS WATER, EMBOSS STRETCHER, EMBOSS NEEDLE, EMBOSS LALIGN, BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software. Those skilled in this art can determine the appropriate parameters used to measure the degree of alignment, including any algorithm required to achieve maximum alignment with the full length of the sequences being compared.

[0074] The terms "cancer" and "tumor" are used interchangeably herein and encompass all types of tumorigenic processes and / or cancerous growth. In embodiments, cancer includes primary tumors as well as metastatic tissues or malignant cells, tissues, or organs. In embodiments, cancer encompasses all histopathological stages, such as stages of cancer invasiveness / severity. In embodiments, cancer includes recurrent and / or resistant cancer.

[0075] As used herein, "treatment" (and its grammatical variations such as "treat" or "treating") refers to a clinical intervention aimed at altering the natural course of disease in the treated individual, and can be aimed at prevention or at addressing the clinical pathological course of the disease. The desired therapeutic effects include, but are not limited to, preventing the onset or recurrence of disease, relieving symptoms, mitigating any direct or indirect pathological consequences of the disease, preventing metastasis, slowing the rate of disease progression, improving or alleviating the disease condition, and alleviating or improving prognosis. In some embodiments, the molecules of this invention are intended to delay disease development or slow disease progression.

[0076] As used in this article, the "ideal weight" ("IBW") is 50 kg + 2.3 kg × (actual height – 60 in) (male) and 45.5 kg + 2.3 kg × (actual height – 60 in) (female).

[0077] As used in this article, "adjusted weight" ("AJBW") is IBW + 0.4 × (actual weight – IBW). [anti] [CD46] [Recombinant Antibody]

[0078] In some embodiments, this document discloses a recombinant antibody (or its antigen-binding fragment) that specifically binds to CD46. In some embodiments, the antibody, its antigen-binding fragment, or a variant monoclonal antibody. In some embodiments, the antibody, its antigen-binding fragment, or a variant human antibody, murine antibody, humanized antibody, or chimeric antibody. In some embodiments, the antibody comprises or consists of a functional fragment of a full-length antibody (e.g., an antigen-binding fragment of a full-length antibody), such as a monovalent Fab, a bivalent Fab'2, a single-chain variable fragment (scFv), or a functional fragment or variant thereof. In some embodiments, the recombinant antibody (or its antigen-binding fragment) comprises an immunoglobulin variable heavy chain domain (VH). In some embodiments, the recombinant antibody (or its antigen-binding fragment) comprises an immunoglobulin variable light chain domain (VL). In some embodiments, the recombinant antibody (or its antigen-binding fragment) comprises both VH and VL.

[0079] In some embodiments, the recombinant antibody (or its antigen-binding fragment) comprises an Fc region. In some embodiments, the recombinant antibody (or its antigen-binding fragment) is a full-length antibody. In some embodiments, the recombinant antibody (or its antigen-binding fragment) comprises a first light chain including a light chain variable region and a light chain constant region; a first heavy chain including a heavy chain variable region and a heavy chain constant region; a second light chain including a light chain variable region and a light chain constant region; and a second heavy chain including a heavy chain variable region and a heavy chain constant region. In some embodiments, the first and second light chains have at least 95%, 96%, 97%, 98%, 99%, or 100% sequence identity. In some embodiments, the first and second light chains bind the same antigenic determinant. In some embodiments, the first and second heavy chains have at least 95%, 96%, 97%, 98%, 99%, or 100% sequence identity. In some embodiments, the first and second heavy chains bind the same antigenic determinant.

[0080] In some embodiments, the recombinant antibody (or its antigen-binding fragment) is derived from a non-human antibody (e.g., rabbit or mouse). In some cases, the humanized form of the non-human antibody contains minimal non-human sequence to maintain the original antigen specificity. In some cases, the humanized antibody system is a human immunoglobulin (receptor antibody), wherein the CDR of the receptor antibody is replaced by residues of the CDR of a non-human immunoglobulin (donor antibody), such as rat, rabbit, or mouse donors having the desired specificity, affinity, binding, binding kinetics, and / or capacity. In some cases, one or more framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues of the donor antibody. Complementarity Determinant Region (CDR)

[0081] In some embodiments, the recombinant antibody binding to CD46 comprises an immunoglobulin variable heavy chain domain (VH) which includes at least one, two, or three complementarity-determining regions (CDRs) disclosed in Table 1 or sequences substantially identical thereto (e.g., sequences having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).

[0082] In some embodiments, the recombinant antibody binding to CD46 comprises an immunoglobulin variable light chain domain (VL) which includes at least one, two, or three complementarity-determining regions (CDRs) disclosed in Table 2 or sequences substantially identical thereto (e.g., sequences having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).

[0083] In some embodiments, the recombinant antibody binding to CD46 comprises VH, which contains at least one, two, or three complementarity-determining regions (CDRs) disclosed in Table 1 or sequences substantially identical thereto (e.g., sequences having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity); and VL, which contains at least one, two, or three complementarity-determining regions (CDRs) disclosed in Table 2 or sequences substantially identical thereto (e.g., sequences having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).

[0084] In some embodiments, the recombinant antibody binding to CD46 comprises VH, which includes CDR1 of SEQ ID NO: 1, CDR2 of SEQ ID NO: 2, and CDR3 of SEQ ID NO: 3.

[0085] In some embodiments, the recombinant antibody binding to CD46 comprises VL, which includes CDR1 of SEQ ID NO: 4, CDR2 of SEQ ID NO: 5, and CDR3 of SEQ ID NO: 6.

[0086] In some embodiments, the recombinant antibody binding to CD46 comprises VH, which comprises CDR1 of SEQ ID NO: 1, CDR2 of SEQ ID NO: 2, and CDR3 of SEQ ID NO: 3; and VL, which comprises CDR1 of SEQ ID NO: 4, CDR2 of SEQ ID NO: 5, and CDR3 of SEQ ID NO: 6. [surface] [1.] [like] [Kabat] [Resistance as defined by others] [CD46] [Antibodies] [VH CDR] [Amino acid sequence] [.] [Antibody] [SEQ ID NO] [CDR1] [SEQ ID NO] [CDR2] [SEQ ID NO] [CDR3] YS5FL 1 GLTVNNYA 2 ISYDGNNK 3 AKGGGYFDL [surface] [2.] [like] [Kabat] [Resistance as defined by others] [CD46] [Antibodies] [VL CDR] [Amino acid sequence] [.] [Antibody] [SEQ ID NO] [CDR1] [SEQ ID NO] [CDR2] [SEQ ID NO] [CDR3] YS5FL 4 SSNIGAGYD 5 GNN 6 SSYTSGTWL

[0087] In some embodiments, the CDR described herein comprises one, two, or three amino acid modifications. In some embodiments, the modification is substitution, addition, or deletion. In some embodiments, the CDR described herein comprises one, two, or three retained amino acid substitutions. In some embodiments, the one, two, or three amino acid modifications do not substantially alter the binding to human CD46. In some embodiments, the one, two, or three amino acid modifications alter the binding to human CD46. In some embodiments, VH-CDR3 and / or VL-CDR3 comprise amino acid substitutions that alter the binding to human CD46, immunogenicity, or some other characteristic. In some embodiments, the amino acid substitute is alanine (A). Variable heavy chain region and variable light chain region

[0088] In some embodiments, the recombinant antibody binding to CD46 comprises VH, which comprises the amino acid sequence disclosed in Table 3 or a sequence substantially identical thereto (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).

[0089] In some embodiments, the recombinant antibody binding to CD46 comprises VL, which comprises the amino acid sequence disclosed in Table 4 or a sequence substantially identical thereto (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).

[0090] In some embodiments, the recombinant antibody binding to CD46 comprises VH, which comprises the amino acid sequence disclosed in Table 3 or a sequence substantially identical thereto (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity); and VL, which comprises the amino acid sequence disclosed in Table 4 or a sequence substantially identical thereto (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).

[0091] In some embodiments, the recombinant antibody binding to CD46 comprises VH, which comprises the amino acid sequence of SEQ ID NO: 7 or a sequence substantially identical thereto (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).

[0092] In some embodiments, the recombinant antibody binding to CD46 comprises VL, which comprises the amino acid sequence of SEQ ID NO: 8 or a sequence substantially identical thereto (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).

[0093] In some embodiments, the recombinant antibody binding to CD46 comprises VH, which comprises the amino acid sequence of SEQ ID NO: 7 or a sequence substantially identical thereto (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity); and VL, which comprises the amino acid sequence of SEQ ID NO: 8 or a sequence substantially identical thereto (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity). [surface] [3.] Amino acid sequence of the anti-CD46 variable heavy chain binding domain. [name] [SEQ ID NO] [Amino acid sequence] YS5FL 7 QVQLVQSGGGVVQPGRSLRLACAASGLTVNNYAMHWVRQAPGKGLEWVAVISYDGNNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGGGYFDLWGRGTLVTVSS [surface] [4.] Amino acid sequence of the anti-CD46 variable light chain binding domain. [name] [SEQ ID NO] [Amino acid sequence] YS5FL 8 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNNNRPSGVPDRFSGSKTSASLAITGLQAEDEADYYCSSYTSGTWLFGGGTKLTVL Heavy chains and light chains

[0094] In some embodiments, the recombinant antibody binding to CD46 comprises a heavy chain that includes the amino acid sequences disclosed in Table 5 or sequences substantially identical thereto (e.g., sequences having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).

[0095] In some embodiments, the recombinant antibody binding to CD46 comprises a light chain containing the amino acid sequences disclosed in Table 6 or sequences substantially identical thereto (e.g., sequences having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).

[0096] In some embodiments, the recombinant antibody binding to CD46 comprises a heavy chain comprising an amino acid sequence disclosed in Table 5 or a sequence substantially identical thereto (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity); and a light chain comprising an amino acid sequence disclosed in Table 6 or a sequence substantially identical thereto (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).

[0097] In some embodiments, the recombinant antibody binding to CD46 comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 9 or a sequence substantially identical thereto (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).

[0098] In some embodiments, the recombinant antibody binding to CD46 comprises a light chain containing the amino acid sequence of SEQ ID NO: 10 or a sequence substantially identical thereto (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).

[0099] In some embodiments, the recombinant antibody binding to CD46 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 9 or a sequence substantially identical thereto (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity); and a light chain comprising the amino acid sequence of SEQ ID NO: 10 or a sequence substantially identical thereto (e.g., a sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity). [, , ] [] [surface] [5.] Anti-CD46 heavy chain amino acid sequence. [name] [SEQ ID NO] [Amino acid sequence] YS5FL 9 QVQLVQSGGGVVQPGRSLRLACAASGLTVNNYAMHWVRQAPGKGLEWVAVISYDGNNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGGGYFDLWGRGTL VTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTH TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK [surface] [6.] Anti-CD46 light chain amino acid sequence. [name] [SEQ ID NO] [Amino acid sequence] YS5FL 10 QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYGNNNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCSSYTSGTWLFGGGTKLT VLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS

[0100] In some embodiments, the anti-CD46 antibody disclosed herein comprises an immunoglobulin constant region (e.g., an Fc region). An exemplary Fc region may be selected from the heavy chain constant region of IgG1, IgG2, IgG3, or IgG4; more specifically, the heavy chain constant region of human IgG1 or IgG4. In some embodiments, the immunoglobulin constant region (e.g., the Fc region) is altered (e.g., mutated) to enhance or degrade one or more of the following: Fc receptor binding, antibody glycosylation, number of cysteine ​​residues, effector cell function, or complement function. [Effective agent]

[0101] In some embodiments, this document discloses an immune conjugate comprising an anti-CD46 antibody attached to an effector (or its prodrug). In some embodiments, the effector is a drug (or its prodrug), a small molecule, a protein, a peptide, an antibody, a ligand, a receptor, a cytotoxic agent, a cell inhibitor, a liposome, a nanoparticle, a radioactive nucleus, a cytokine, a chemokine, a toxin, a detectable marker, a viral particle, or a chelating agent.

[0102] In some embodiments, the effector is a drug (or its prodrug). In some embodiments, the effector is an anticancer agent (or its prodrug). In some embodiments, the effector is a chemotherapeutic agent (or its prodrug). In some embodiments, the effector is a microtubule inhibitor (or its prodrug), a DNA disruptor (or its prodrug), or a polymerase inhibitor (or its prodrug).

[0103] In some embodiments, the effector is a microtubule inhibitor (or its prodrug). In some embodiments, the microtubule inhibitor is auristatin (or a derivative thereof), dolastatin-10 (or a derivative thereof), or maytansine (or a derivative thereof). In some embodiments, the microtubule inhibitor is monomethylauristatin F (MMAF), auristatin E (AE), monomethylauristatin E (MMAE), valine-citrulline MMAE (vcMMAE), or valine-citrulline MMAF (vcMMAF). In some embodiments, the microtubule inhibitor is monomethylauristatin E (MMAE).

[0104] In some embodiments, the effector comprises or consists of a compound of formula A: (Formula A) Molecular formula: C 39H 67N 5O 7

[0105] In certain embodiments, the effector comprises a detectable marker. Suitable detectable markers include (but are not limited to) radiopaque markers, nanoparticles, PET markers, MRI markers, radioactive markers, and similar markers. In radioactive nuclei and in various embodiments of the invention, gamma emitters, positron emitters, X-ray emitters, and fluorescent emission systems are suitable for localization, diagnosis, and / or grading and / or treatment, while beta and alpha emitters, as well as electron and neutron trapping agents (such as boron and uranium), can also be used for treatment. [Immune conjugates]

[0106] In one embodiment, this document provides an immune conjugate comprising an anti-CD46 antibody and an effector. In some embodiments, the methods described herein employ such immune conjugates.

[0107] In some embodiments, the immune conjugate comprises an anti-CD46 antibody (or an antigen-binding fragment thereof) as described herein. In some embodiments, the immune conjugate comprises a YS5FL antibody (or an antigen-binding fragment thereof).

[0108] In some embodiments, the effector binds to the anti-CD46 antibody. In some embodiments, the effector is attached to the anti-CD46 antibody via a linker. In some embodiments, the linker is a peptide linker, a small molecule linker, or a linker comprising a peptide and a small molecule. Exemplary peptide linkers include (but are not limited to) peptide linkers comprising glycine, serine, or a combination of glycine and serine.

[0109] In some embodiments, the linker is cleavable. In some embodiments, the linker cleaves only upon internalization into cells. In some embodiments, the cleavable linker cleaves only upon internalization into cancer cells. In some embodiments, the cleavable portion of the linker is a peptide (e.g., a dipeptide, such as ValCit). In some embodiments, the cleavable linker is cleaved by cathepsins. In some embodiments, the linker comprises maleimide. In some embodiments, the linker comprises lanolinic acid. In some embodiments, the linker comprises maleimide and lanolinic acid. In some embodiments, the linker comprises maleimide, lanolinic acid, and a cleavable dipeptide.

[0110] In some embodiments, the linker comprises or is composed of maleiminohexyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB).

[0111] In some embodiments, the connector comprises or is composed of a compound of formula B: (Formula B)

[0112] In some embodiments, the effector is attached to the light chain of the anti-CD46 antibody. In some embodiments, the effector is attached to the constant region of the light chain of the anti-CD46 antibody. In some embodiments, the effector is attached to the heavy chain of the anti-CD46 antibody. In some embodiments, the effector is attached to the constant region of the heavy chain of the anti-CD46 antibody.

[0113] In some embodiments, the effector moiety is attached to the cysteine ​​residues of the anti-CD46 antibody. In some embodiments, the anti-CD46 antibody system is partially reduced prior to binding to the effector moiety to reduce 1-4 interchain disulfide bonds, without reducing intrachain disulfide bonds. Partial reduction exposes cysteine ​​residue pairs, making them available for binding to adducts such as mc-vc-PAB-MMAE. In some embodiments, the following interchain cysteine ​​pairs of YS5FL are exposed: C219 of the first heavy chain and C214 of the first light chain; C219 of the second heavy chain and C214 of the second light chain; C225 of the first heavy chain and C225 of the second light chain; and C228 of the first heavy chain and C228 of the second light chain. In some embodiments, the effector, such as mc-vc-PAB-MMAE, binds to 0, 1, 2, 3, or 4 pairs of cysteine ​​residues on YS5FL.

[0114] In some embodiments, the ratio of effector to anti-CD46 antibody is c. In some embodiments, the ratio of effector to anti-CD46 antibody is 2:1, 4:1, 6:1, or 8:1. In some embodiments, the ratio of effector to anti-CD46 antibody is about 4:1. In some embodiments, the average ratio of effector to anti-CD46 antibody is about 3.7:1. In some embodiments, if the immune conjugate contains two or more effectors, then all effectors are identical. In some embodiments, if the immune conjugate contains two or more effectors, then at least two effectors are different. In some embodiments, the ratio of effector to anti-CD46 antibody is about 4:1, and all effectors are identical. Exemplary immunoconjugates

[0115] The exemplary immune conjugates provided herein comprise an anti-CD46 YS5FL antibody linked to a monomethylaurestatin E (MMAE) effector via maleiminohexylacetyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB). In some embodiments, the ratio of MMAE to YSFL antibody is approximately 4:1.

[0116] In some embodiments, the immunoconjugate comprises an antibody conjugate of Formula C below, wherein it comprises the heavy chain of SEQ ID NO: 9 and the light chain of SEQ ID NO: 10. This immunoconjugate is also referred to herein as FOR46 and comprises a YS5FL antibody attached to MMAE via an mc-vc-PAB linker. Formula C

[0117] In some embodiments, the anti-CD46 immunoconjugate described herein is manufactured via a process comprising a disulfide bond of reduced or partially reduced immunoglobulin. In some embodiments, the anti-CD46 immunoconjugate described herein is manufactured via a process comprising an interchain disulfide bond of reduced or partially reduced immunoglobulin. In some embodiments, the reducing agent is dithiothreitol (DTT) or tris(2-carboxyethyl)phosphine (TCEP). In some embodiments, the effector-linker complex comprising a maleimine reactive group binds to a reduced cysteine ​​pair of immunoglobulin. In some embodiments, the effector-linker complex is mc-vc-PAB-MMAE.

[0118] In some embodiments, the effector-linker complex is bound to C219, C225, or C228 of the YS5FL heavy chain (SEQ ID NO: 9) or C214 of the YS5FL light chain (SEQ ID NO: 10), or any combination thereof. In some embodiments, the effector-linker complex is bound to C219 of the YS5FL heavy chain and C214 of the YS5FL light chain. In some embodiments, the anti-CD46 immunoconjugate comprises two YS5FL heavy chains and two YS5FL light chains, and the effector-linker complex is bound to C219 of the first YS5FL heavy chain, C214 of the first YS5FL light chain, C219 of the second YS5FL heavy chain, and C214 of the second YS5FL light chain. In some embodiments, the anti-CD46 immunoconjugate comprises two YS5FL heavy chains, and the effector-linker complex is bound to C225 of the first YS5FL heavy chain and C225 of the second YS5FL heavy chain. In some embodiments, the anti-CD46 immune conjugate comprises two YS5FL heavy chains, and the effector-conjugate complex is bound to C228 of the first YS5FL heavy chain and C228 of the second YS5FL heavy chain. In some embodiments, the immune conjugate comprises two, four, six, or eight effectors, and the effectors are bound to any one, two, three, or four of the following cysteine ​​pairs: C219 of HC1 and C214 of LC1; C219 of HC2 and C214 of LC2; C225 of HC1 and C225 of HC2; and C228 of HC1 and C228 of HC2. Immune conjugates that bind to target cells and their activity against target cells

[0119] In some embodiments, the anti-CD46 antibody or immunoconjugate described herein binds to and is internalized by CD46 expressed on the surface of target cells (e.g., cancer cells). In some embodiments, the antibody or immunoconjugate is internalized into the target cell via macropinocytosis. In some embodiments, the antibody or immunoconjugate targets the lysosomes of the cell upon internalization. In some embodiments, the antibody or immunoconjugate initiates internalization into the cell without cross-linking.

[0120] In some embodiments, the anti-CD46 antibody or immune conjugate described herein mediates the killing of target cells (e.g., cancer cells) upon internalization. In some embodiments, the anti-CD46 antibody or immune conjugate induces apoptosis of target cells (e.g., cancer cells) upon internalization. In some embodiments, the anti-CD46 antibody or immune conjugate inhibits cell division of target cells (e.g., cancer cells) upon internalization. In some embodiments, the anti-CD46 antibody or immune conjugate selectively inhibits cell division of cancer cells upon internalization but does not inhibit cell differentiation of non-cancer cells upon internalization. [Manufacturing antibodies or their antigen-binding fragments]

[0121] In some embodiments, the antibody (and its antigen-binding fragment) is manufactured using any method known in this art for synthesizing antibodies, specifically by chemical synthesis or by recombinant expression techniques.

[0122] In some embodiments, the antibody (or its antigen-binding fragment) is expressed in a recombinant manner. In some embodiments, the nucleic acid encoding the antibody (or its antigen-binding fragment) is assembled from chemically synthesized oligonucleotides. In some embodiments, the nucleic acid molecule encoding the antibody is generated from a suitable source (e.g., an antibody cDNA library or cDNA library generated from any tissue or cell expressing immunoglobulins) by PCR amplification using synthetic primers that can confound the 3' and 5' ends of the sequence or by selection using oligonucleotide probes targeting a specific gene sequence.

[0123] In some embodiments, antibodies (or antigen-binding fragments thereof) are produced by immunizing animals such as mice to generate polyclonal or monoclonal antibodies.

[0124] In some embodiments, an expression vector containing the antibody's nucleotide sequence or the antibody's nucleotide sequence is transferred into a host cell using conventional techniques (e.g., electroporation, liposome transfection, and calcium phosphate precipitation), and the transfected cells are subsequently cultured using conventional techniques to produce the antibody. In some embodiments, antibody expression is regulated by constitutive, inducible, or tissue-specific promoters.

[0125] Various host expression vector systems can be used to express the antibodies (or their antigen-binding fragments) described herein. These include (but are not limited to) microorganisms such as: bacteria transformed using recombinant phage DNA, plasmid DNA, or cohesive plasmid DNA expression vectors containing sequences encoding antibodies or their binding fragments (e.g., *Escherichia coli* and *Bacillus subtilis*); and yeast transformed using recombinant yeast expression vectors containing sequences encoding antibodies or their binding fragments (e.g., *Saccharomyces*). Pichia); insect cell systems infected with recombinant viral expression vectors (e.g., baculoviruses) containing sequences encoding antibodies or their binding fragments; plant cell systems infected with recombinant viral expression vectors (e.g., cauliflower mosaic virus (CaMV) and tobacco mosaic virus (TMV)) or transformed with recombinant plastid expression vectors (e.g., Ti plastids) containing sequences encoding antibodies or their binding fragments; or mammalian cell systems containing recombinant expression constructs containing promoters derived from mammalian cell genomes (e.g., metallothionein promoters) or promoters derived from mammalian viruses (e.g., adenovirus late promoters; vaccinia virus 7.5K promoters) (e.g., COS, CHO, BH, 293, 293T, 3T3 cells).

[0126] For long-term, high-yield production of recombinant proteins, stable performance is preferred. In some embodiments, cell lines with stable antibody performance are manufactured. After introducing foreign DNA, the engineered cells are grown in a concentrated medium for 1-2 days, and then converted to a selective medium. Selectable markers in the recombinant plasmids can be used to selectively confer resistance.

[0127] In some embodiments, any method known in this technology for purifying antibodies may be used, such as chromatography (e.g., ion exchange, affinity (especially affinity of protein A for a specific antigen) and particle size column chromatography), centrifugation, differential dissolution, or purification by any other standard technique for purifying proteins. [Vessel]

[0128] Vectors may include any suitable vector derived from eukaryotic or prokaryotic sources. In some cases, vectors are derived from bacteria (e.g., *Escherichia coli*), insects, yeasts (e.g., *Pichia pastoris*), algae, or mammalian sources. Exemplary bacterial vectors include pACYC177, pASK75, the pBAD vector series, the pBADM vector series, the pET vector series, the pETM vector series, the pGEX vector series, pHAT, pHAT2, pMal-c2, pMal-p2, the pQE vector series, pRSET A, pRSET B, pRSET C, the pTrcHis2 series, pZA31-Luc, pZE21-MCS-1, pFLAG ATS, pFLAG CTS, pFLAG MAC, pFLAG Shift-12c, pTAC-MAT-1, pFLAG CTC, or pTAC-MAT-2.

[0129] Exemplary insect vectors include pFastBac1, pFastBac DUAL, pFastBac ET, pFastBac HTa, pFastBac HTb, pFastBac HTc, pFastBac M30a, pFastBact M30b, pFastBac, M30c, pVL1392, pVL1393, pVL1393 M10, pVL1393 M11, pVL1393 M12, FLAG vectors such as pPolh-FLAG1 or pPolh-FLAG2, or MAT vectors such as pPolh-MAT1 or pPolh-MAT2.

[0130] In some cases, yeast vectors include Gateway® pDEST™ 14 vector, Gateway® pDEST™ 15 vector, Gateway® pDEST™ 17 vector, Gateway® pDEST™ 24 vector, Gateway® pYES-DEST52 vector, pBAD-DEST49 Gateway® target vector, pAO815 Pichia pastoris vector, pFLD1 Pichia pastoris vector, pGAPZA, B and C Pichia pastoris vectors, pPIC3.5K Pichia pastoris vector, pPIC6 A, B and C Pichia pastoris vector, pPIC9K Pichia pastoris vector, pTEF1 / Zeo, pYES2 yeast vector, pYES2 / CT yeast vector, pYES2 / NT A, B and C yeast vector, or pYES3 / CT yeast vector.

[0131] Exemplary algal vectors include pChlamy-4 vector or MCS vector.

[0132] Examples of mammalian vectors include transient or stable expression vectors. Transient mammalian expression vectors may include pRK5, p3xFLAG-CMV 8, pFLAG-Myc-CMV 19, pFLAG-Myc-CMV 23, pFLAG-CMV 2, pFLAG-CMV 6a, b, c, pFLAG-CMV 5.1, pFLAG-CMV 5a, b, c, p3xFLAG-CMV 7.1, pFLAG-CMV 20, p3xFLAG-Myc-CMV 24, pCMV-FLAG-MAT1, pCMV-FLAG-MAT2, pBICEP-CMV 3, or pBICEP-CMV 4. Stable mammalian expression vectors may include pFLAG-CMV 3, p3xFLAG-CMV 9, p3xFLAG-CMV 13, pFLAG-Myc-CMV 21, p3xFLAG-Myc-CMV 25, pFLAG-CMV 4, p3xFLAG-CMV 10, p3xFLAG-CMV 14, pFLAG-Myc-CMV 22, p3xFLAG-Myc-CMV 26, pBICEP-CMV 1, or pBICEP-CMV 2.

[0133] In some cases, cell-free systems are mixtures of cytoplasmic and / or nuclear components derived from cells and are used for in vitro nucleic acid synthesis. In other cases, cell-free systems utilize prokaryotic or eukaryotic cell components. Typically, nucleic acid synthesis is achieved in cell-free systems based on, for example, Drosophila cells, Xenopus eggs, or HeLa cells. Exemplary cell-free systems include (but are not limited to) the E. coli S30 extraction system, the E. coli T7 S30 system, or PURExpress®. [Host cell]

[0134] The host cell can be any suitable cell, such as naturally derived cells or genetically modified cells. In some cases, the host cell line produces the host cell. In some cases, the host cell line is a eukaryotic cell. In others, the host cell line is a prokaryotic cell. In some cases, eukaryotic cells include fungi (e.g., yeast cells), animal cells, or plant cells. In some cases, prokaryotic cells are bacterial cells. Examples of bacterial cells include Gram-positive bacteria or Gram-negative bacteria. Gram-negative bacteria are often anaerobic, rod-shaped, or both.

[0135] In some cases, Gram-positive bacteria include Actinomycetes, Firmicutes, or Soft-walled Bacteria. In other cases, Gram-negative bacteria include Aquagenic Bacteria, Radiation-resistant *Hematoxylin and Fibrobacteria*-*Aquatic Thermophila*, Fibrobacteria-*Aureobacteria* / *Bacteroidetes* (Group FCB), Fusobacteria, *Bacillus*, *Nitrospirillum*, *Planctomyces*-*Verrucous / *Chlamydia* (Group PVC), Proteobacteria, Spirochetes, or Mutual Trophobes. Other bacteria may be Acidobacteria, *Curvoidobacteria*, *Aureobacteria*, *Cyanobacteria*, *Deferobacteria*, *Dendrobacteria*, *Heat Desulfurization Bacteria*, or *Heat Coliforms*. Bacterial cells may be *Escherichia coli*, *Clostridium botulinum*, or *Escherichia coli*.

[0136] Exemplary prokaryotic host cells include (but are not limited to) BL21, Mach1™, DH10B™, TOP10, DH5α, DH10Bac™, OmniMax™, MegaX™, DH12S™, INV110, TOP10F', INVαF, TOP10 / P3, ccdB Survival, PIR1, PIR2, Stbl2™, Stbl3™, or Stbl4™.

[0137] In some cases, animal cells include cells derived from vertebrates or invertebrates. In others, animal cells include cells derived from marine invertebrates, fish, insects, amphibians, reptiles, or mammals. In still others, fungal cells include yeast cells, such as brewer's yeast, baker's yeast, or wine yeast.

[0138] Fungi include the class Ascomycota, such as yeasts, molds, filamentous fungi, Basidiomycetes, or Zygomycetes. In some cases, yeasts include either Ascomycota or Basidiomycetes. In some cases, Ascomycota includes the subphylum Yeast (true yeasts, such as *Saccharomyces cerevisiae*, baker's yeast) or the subphylum Exomycota (such as *Schizosaccharomyces*). In some cases, Basidiomycetes includes the class Agaricales (such as *Tremella fuciformis*) or the subphylum Stylorhynchoides (such as *Ustilago maydis*).

[0139] Exemplary yeasts or filamentous fungi include, for example, the following genera: *Saccharomyces*, *Schizosaccharomyces*, *Candida*, *Pichia*, *Hansenula*, *Kluyveromyces*, *Zygosaccharomyces*, *Yersinia*, *Hydrospermia*, *Rhodotorula*, *Aspergillus*, *Fusarium*, or *Trichoderma*. Examples of yeasts or filamentous fungi include, for example, the following species: *Saccharomyces cerevisiae*, *Schizosaccharomyces cerevisiae*, *Candida utilis*, *Candida boidini*, *Candida albicans*, *Candida tropicalis*, *Candida asteroides*, *Candida glabrata*, *Candida krusei*, *Candida parapsilosis*, *Candida guilliermondii*, *Candida viswanathii*, *Candida Portugueseia*, *Rhodotorula glutinis*, *Pichia metanolica*, *Pichia angusta*, *Pichia pastoris*, *Pichia aberrant*, *Hansenula polymorpha*, *Kluyveromyces lactis*, *Zygosaccharomyces rouxii*, and *Yarrowia lipolytica*. lipolytica), spore yeast, red thallus yeast-black yeast, small nest mold, Aspergillus avocado, green yeast, Trichoderma reesei, Yersinia lipolytica, Brettanomyces bruxellensis, Candida astrum, Schizosaccharomyces nigra, Torulaspora delbrueckii, Zygosaccharomyces bailii, Cryptococcus neoformans, Cryptococcus gattii, or Saccharomyces boulardii.

[0140] Exemplary yeast host cells include (but are not limited to) Pichia pastoris yeast strains such as GS115, KM71H, SMD1168, SMD1168H and X-33; and Saccharomyces cerevisiae yeast strains such as INVSC1.

[0141] In some cases, other animal cells include cells derived from mollusks, arthropods, annelids, or sponges. In some cases, other animal cell lines are mammalian cells, such as cells from primates, apes, horses, cattle, pigs, dogs, cats, or mice. In some cases, mice include mice, rats, hamsters, gerbils, chinchillas, fancy rats, or guinea pigs.

[0142] Exemplary mammalian host cells include (but are not limited to) 293A cell line, 293FT cell line, 293F cell line, 293H cell line, CHO DG44 cell line, CHO-S cell line, CHO-K1 cell line, FUT8 KO CHOK1 cell line, Expi293F™ cell line, Flp-In™ T-REx™ 293 cell line, Flp-In™-293 cell line, Flp-In™-3T3 cell line, Flp-In™-BHK cell line, Flp-In™-CHO cell line, Flp-In™-CV-1 cell line, Flp-In™-Jurkat cell line, FreeStyle™ 293-F cell line, FreeStyle™ CHO-S cell line, GripTite™ 293 MSR cell line, GS-CHO cell line, HepaRG™ cell line, and T-REx™ cell line. Jurkat cell line, Per.C6 cell line, T-REx™-293 cell line, T-REx™-CHO cell line and T-REx™-HeLa cell line.

[0143] In some cases, mammalian host cell lines are stable cell lines or cell lines that have incorporated relevant genetic material into their own genome and have the ability to express the products of genetic material after many generations of cell division. In other cases, mammalian host cell lines are variable cell lines or cell lines that have not incorporated relevant genetic material into their own genome and do not have the ability to express the products of genetic material after many generations of cell division.

[0144] Exemplary insect host cells include (but are not limited to) Drosophila S2 cells, Sf9 cells, Sf21 cells, High Five™ cells, and expresSF+® cells.

[0145] In some cases, plant cells include cells derived from algae. Exemplary plant cell lines include (but are not limited to) those derived from *Chlamydomonas reinhardtii* 137c or *Synechococcus slenderus* PPC 7942. [Treatment methods]

[0146] In one instance, this article provides a method for treating cancer by administering an anti-CD46 antibody or immune conjugate as described herein.

[0147] In some embodiments, the cancer is multiple myeloma. In some embodiments, the cancer is relapsed multiple myeloma. In some embodiments, the cancer is remission-associated multiple myeloma. In some embodiments, the cancer is either relapsed or remission-associated multiple myeloma.

[0148] In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is castration-resistant prostate cancer. In some embodiments, the cancer is metastatic prostate cancer.

[0149] In one embodiment, this document provides an anti-CD46 antibody or immune conjugate as described herein for use as a pharmaceutical product. In another embodiment, this document provides an anti-CD46 antibody or immune conjugate as described herein for treating a disease, particularly for treating cancer. In another embodiment, this document provides an anti-CD46 antibody or immune conjugate as described herein for a method of treating cancer. In another embodiment, this document provides an anti-CD46 antibody or immune conjugate as described herein for treating a disease in an individual in need. In another embodiment, this document provides an anti-CD46 antibody or immune conjugate as described herein for a method of treating an individual with cancer, the method comprising administering a therapeutically effective amount of the anti-CD46 antibody or immune conjugate as described herein to the individual. In another embodiment, this document provides an anti-CD46 antibody or immune conjugate as previously described herein in the manufacture or preparation of a pharmaceutical product for treating a disease in an individual in need. In another embodiment, this document provides a pharmaceutical product for a method of treating cancer, the method comprising administering a therapeutically effective amount of the pharmaceutical product to an individual with cancer. Drug administration and delivery

[0150] The anti-CD46 antibodies or immune conjugates described herein can be formulated, administered, and given in accordance with good medical practice for use in treatment. In this case, factors to consider include the specific disease being treated, the specific mammal being treated, the individual patient's clinical condition, the cause of the disease, the site of delivery, the method of administration, the timing of administration, and other factors known to the medical practitioner.

[0151] In some embodiments, the antibodies or immune conjugates described herein are administered to human individuals via intravenous infusion. In some embodiments, the antibodies or immune conjugates are administered to human individuals every 7 days, every 14 days, every 18 days, every 21 days, or every 30 days. In some embodiments, the antibodies or immune conjugates are administered to human individuals every 21 days.

[0152] In some embodiments, the antibody or immune conjugate is administered to a human individual at a dose of about 1.0 to about 5.0 mg / kg. In some embodiments, the dosage of the antibody or immune conjugate is about 1.0 to about 4.5 mg / kg, about 1.0 to about 4.0 mg / kg, about 1.0 to about 3.5 mg / kg, about 1.0 to about 3.0 mg / kg, about 1.0 to about 2.7 mg / kg, about 1.0 to about 2.5 mg / kg, about 1.0 to about 2.4 mg / kg, about 1.5 to about 4.5 mg / kg, about 1.5 to about 4.0 mg / kg, about 1.5 to about 3.5 mg / kg, about 1.5 to about 3.0 mg / kg, about 1.5 to about 2.7 mg / kg, about 1.5 to about 2.5 mg / kg, about 1.5 to about 2.4 mg / kg, about 1.5 to about 2.0 mg / kg, about 1.8 to about 4.5 mg / kg, about 1.8 to about 4.0 mg / kg, about 1.8 to about 3.5 mg / kg, about 1.8 to about 3.0 mg / kg, about 1.8 to about 2.7 mg / kg. mg / kg, about 1.8 to about 2.5 mg / kg, about 1.8 to about 2.4 mg / kg, or about 1.8 to about 2.0 mg / kg. In some embodiments, the antibody or immune conjugate is administered to a human individual at a dose of about 1.5 to about 2.5 mg / kg. In some embodiments, the antibody or immune conjugate is administered to a human individual at a dose of about 1.2 to about 3.0 mg / kg.

[0153] In some embodiments, the antibody or immunoconjugate is administered to a human individual at a dose of about 1.0, about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2.0, about 2.1, about 2.2, about 2.3, about 2.4, about 2.5, about 2.6, about 2.7, about 2.8, about 2.9, about 3.0, about 3.1, about 3.2, about 3.3, about 3.4, about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, or about 4.0 mg / kg. In some embodiments, the antibody or immunoconjugate is administered to a human individual at a dose of about 1.8, about 2.4, about 2.7, about 3.0, or about 3.2 mg / kg. In some embodiments, the antibody or immunoconjugate is administered to a human individual at a dose of about 1.8 mg / kg. In some embodiments, the antibody or immunoconjugate is administered to a human individual at a dose of about 2.4 mg / kg. In some embodiments, the antibody or immunoconjugate is administered to a human individual at a dose of about 2.7 mg / kg. In some embodiments, the antibody or immunoconjugate is administered to a human individual at a dose of about 3.0 mg / kg. In some embodiments, the antibody or immunoconjugate is administered to a human individual at a dose of about 3.2 mg / kg. In some embodiments, the antibody or immunoconjugate is administered to a human individual at a dose of about 1.5 mg / kg. In some embodiments, the antibody or immunoconjugate is administered to a human individual at a dose of about 2.5 mg / kg. In some embodiments, the antibody or immunoconjugate is administered to a human individual at a dose of about 3.0 mg / kg. In some embodiments, weight is measured in kg. In some embodiments, the weight of the human individual is actual body weight. In some embodiments, weight is measured in kg. In some embodiments, the weight of the human individual is adjusted body weight (AJBW). [Measurement] [CD46] [Performance]

[0154] In one embodiment, this document provides a method for treating cancer in an individual by: (1) determining that the cancer contains CD46, and (2) administering an anti-CD46 antibody or immune conjugate as described herein. In some embodiments, the cancer expressing CD46 is sensitive to treatment with the anti-CD46 antibody or immune conjugate. In some embodiments, the anti-CD46 antibody or immune conjugate is a more effective anticancer agent when the cancer expresses CD46 or expresses a higher level of CD46 compared to a non-cancerous control. In some embodiments, the non-cancerous control is a matched non-cancerous control tissue from a subject / individual who does not have cancer. For example, if the cancer is prostate cancer, the non-cancerous control tissue could be a healthy prostate.

[0155] In some embodiments, the anti-CD46 anti-system is used to determine CD46 expression in cancer. CD46 expression in cancer (e.g., cancer cells, cancerous lesions, metastatic cells) can be detected by various methods such as immunofluorescence microscopy, immunohistochemistry, or flow cytometry.

[0156] In another embodiment, the copy number of the CD46 gene is determined in cancer. The CD46 gene is located on the q arm (1q32) of chromosome 1 at position 32. In some embodiments, 1q amplification indicates a higher degree of CD46 expression. In some embodiments, 1q amplification includes amplification of 1q32. In some embodiments, 1q amplification includes amplification of 1q21, and the amplification of 1q32 is derived from the amplification of 1q21. In some embodiments, gene amplification includes an increase in the copy number of the CD46 gene. In some embodiments, the copy number of the CD46 gene is 3 or greater. In some embodiments, the copy number of the CD46 gene is 4, 5, 6, 7, or 8. [Pharmaceutical compositions and formulations]

[0157] In another embodiment, the present invention provides a pharmaceutical composition comprising an anti-CD46 antibody or immune conjugate as described herein, for example, for any of the treatments described above. In one embodiment, the pharmaceutical composition comprises an anti-CD46 antibody or immune conjugate provided herein and at least one pharmaceutically acceptable excipient. The preparation of the pharmaceutical composition comprising the anti-CD46 antibody or immune conjugate described herein should be known to those skilled in the art according to the present invention, as illustrated in Remington's Pharmaceutical Sciences, 18th edition, Mack Printing Company, 1990, which is incorporated herein by reference.

[0158] In some embodiments, the pharmaceutical composition comprises a buffer. In some embodiments, the buffer comprises histidine. In some embodiments, the pharmaceutical composition comprises about 10 to about 40 mM, about 10 to about 30 mM, or about 10 to about 20 mM of histidine buffer. In some embodiments, the pharmaceutical composition comprises about 10 mM, about 15 mM, about 20 mM, about 25 mM, about 30 mM, about 35 mM, or about 40 mM of histidine buffer. In some embodiments, the pharmaceutical composition comprises about 20 mM of histidine buffer.

[0159] In some embodiments, the pharmaceutical composition comprises a cryoprotectant. In some embodiments, the cryoprotectant comprises a sugar. In some embodiments, the cryoprotectant comprises sucrose or trehalose. In some embodiments, the cryoprotectant comprises sucrose. In some embodiments, the pharmaceutical composition comprises about 4% to about 12%, about 4% to about 11%, about 4% to about 10%, about 4% to about 9%, about 4% to about 8%, about 5% to about 12%, about 5% to about 11%, about 5% to about 10%, about 5% to about 9%, about 5% to about 8%, about 6% to about 12%, about 6% to about 11%, about 6% to about 10%, about 6% to about 9%, about 6% to about 8%, about 7% to about 12%, about 7% to about 11%, about 7% to about 10%, about 7% to about 9%, or about 7% to about 8%. In some embodiments, the pharmaceutical composition comprises about 8% sucrose.

[0160] In some embodiments, the pharmaceutical composition comprises a stabilizer. In some embodiments, the stabilizer prevents denaturation of the recombinant antibody, prevents aggregation of the immune conjugates, or both. In some embodiments, the stabilizer is polysorbate. In some embodiments, the stabilizer is polysorbate 20. In some embodiments, the stabilizer is polysorbate 80. In some embodiments, the pharmaceutical composition comprises about 0.001% to 0.1%, 0.001% to 0.05%, 0.001% to 0.04%, 0.001% to 0.03%, 0.001% to 0.02%, or 0.001% to 0.01% of polysorbate (e.g., polysorbate 80). In some embodiments, the pharmaceutical composition comprises about 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, or 0.1% of polysorbate (e.g., polysorbate 80). In some embodiments, the pharmaceutical composition comprises about 0.01% of polysorbate (e.g., polysorbate 80).

[0161] In some embodiments, the pharmaceutical composition has a pH of about 5.0 to about 7.0. In some embodiments, the pharmaceutical composition has a pH of about 5.0, 5.5, 6.0, 6.5, 7.0, or 7.5. In some embodiments, the pharmaceutical composition has a pH of about 6.0.

[0162] In some embodiments, the pharmaceutical composition contains the anti-CD46 antibody or immune conjugate described herein at concentrations of about 5.0 mg / ml to 15.0 mg / ml, 5.0 mg / ml to 14.0 mg / ml, 5.0 mg / ml to 13.0 mg / ml, 5.0 mg / ml to 12.0 mg / ml, 5.0 mg / ml to 11.0 mg / ml, 5.0 mg / ml to 10.0 mg / ml, 6.0 mg / ml to 15.0 mg / ml, 7.0 mg / ml to 15.0 mg / ml, 8.0 mg / ml to 15.0 mg / ml, 9.0 mg / ml to 15.0 mg / ml, or 10.0 mg / ml to 15.0 mg / ml. In some embodiments, the pharmaceutical composition contains the anti-CD46 antibody or immune conjugate described herein at concentrations of about 5.0 mg / ml, 6.0 mg / ml, 7.0 mg / ml, 8.0 mg / ml, 9.0 mg / ml, 10.0 mg / ml, 11.0 mg / ml, 12.0 mg / ml, 13.0 mg / ml, 14.0 mg / ml, or 15.0 mg / ml. In some embodiments, the pharmaceutical composition contains the anti-CD46 antibody or immune conjugate described herein at concentrations of about 5.0 mg / ml ± 1.0 mg / mL, 6.0 mg / ml ± 1.0 mg / mL, 7.0 mg / ml ± 1.0 mg / mL, 8.0 mg / ml ± 1.0 mg / mL, 9.0 mg / ml ± 1.0 mg / mL, 10.0 mg / ml ± 1.0 mg / mL, 11.0 mg / ml ± 1.0 mg / mL, 12.0 mg / ml ± 1.0 mg / mL, 13.0 mg / ml ± 1.0 mg / mL, 14.0 mg / ml ± 1.0 mg / mL, or 15.0 mg / ml ± 1.0 mg / mL. In some embodiments, the pharmaceutical composition contains the anti-CD46 antibody or immune conjugate described herein at a concentration of about 10.0 mg / ml ± 1.0 mg / mL. Exemplary formulations

[0163] An exemplary formulation of the anti-CD46 antibody or immunoconjugate described herein comprises the anti-CD46 antibody or immunoconjugate described herein at a concentration of approximately 10.0 mg / ml ± 1.0 mg / mL; approximately 20 mM histidine buffer; approximately 8.0% sucrose; approximately 0.01% polysorbate 80; pH 6.0. [Products]

[0164] In another aspect of the invention, an article is provided containing material for treating the cancer described above. The article includes a container and markings or instructions for use on or accompanying the container. Suitable containers include, for example, bottles, vials, syringes, IV solution bags, etc. The container can be formed from various materials such as glass or plastic. The container contains a composition effective for treating the condition, either alone or in combination with another composition, and may have a sterile access port (for example, the container may be an IV solution bag or vial with a stopper that can be punctured by a hypodermic needle).

[0165] The label or package insert indicates that the composition is used to treat a selected condition. Furthermore, the article may comprise (a) a first container containing the composition, wherein the composition contains the bispecific antibody of the present invention; and (b) a second container containing the composition, wherein the composition contains another cytotoxic or other therapeutic agent. The article in this embodiment of the invention may further comprise a package insert indicating that the composition can be used to treat a specific condition.

[0166] Alternatively, the article may further comprise a second (or third) container containing a pharmaceutically acceptable buffer, such as antibacterial water for injection (BWFI), phosphate-buffered saline, Ringer's solution, and dextran solution. It may further include other materials required from a commercial and user's point of view, including other buffers, diluents, filters, needles, and syringes. [Example]

[0167] These examples are provided for illustrative purposes only and do not limit the scope of the patent applications presented herein. [Example] [1] [:] [YS5FL] [Binds on the surface of cancer cells]

[0168] The CD46 cell surface line was detected by flow cytometry. Cells were harvested, centrifuged, and resuspended in FACS buffer (PBS + 2% FBS) at a concentration of 1 × 10⁶ cells / mL. 100 μL of the cell suspension was dispersed in each well of a 96-well plate, and 100 μL of YS5FL was added to each well at 10 μg / mL. The cells were incubated at 4°C for 1 hour. The cell line was washed three times with FACS buffer. After the third wash, the cells were resuspended in 100 μL of Alexa Fluor-488 mouse anti-human IgG1 Fc secondary antibody diluted 1:500 and incubated at 4°C in the dark for 1 hour. The cell line was washed three times with 200 μL PBS at 2000 RPM for 5 minutes. After the final wash, the cells were resuspended in 300 μL of cold PBS and analyzed using a FACSVerse™ (BD Biosciences) flow cytometer. YS5FL specifically binds to the surface of LnCap-C4-2B, LnCap-C4, DU145, PC3-luc, and Hs27 prostate cancer cells, but does not bind to non-tumor BPH1 cells. [picture] [1]. Similarly, YS5FL specifically binds to the surface of RPMI8226, MM.1S, MM.1R and INA6 multiple myeloma cells. [picture] [2]. [Example] [2] [:preparation] [FOR46] [Immune conjugates]

[0169] The structure of YS5FL, which is bonded to the MMAE effector via the mc-vc-PAB linker, is shown in [picture] [3] In the study, purified YS5FL mAb (10 mg / ml) was adjusted to pH 6.8 with sodium phosphate buffer and then treated with TCEP (TCEP / mAb ratio 2.1) at 22°C for two hours. The reduced mAb was reacted with mc-vc-PAB-MMAE (drug / mAb ratio 6) in 9% dimethylacetamide for 15 minutes. The mAb was reduced a second time for one hour, and then conjugated a second time for 60 minutes. The reaction was quenched by lowering the pH to 5.0 with 1M acetic acid to produce a FOR46 immunoconjugate with a drug:antibody ratio of approximately 3.7, as determined by hydrophobic interaction chromatography. [picture] [4] [Example] [3] [:] [FOR46] [Drug Products]

[0170] The FOR46 immune conjugate was formulated into a pharmaceutical product for administration to human individuals. The formulation contained 10.0 ± 1.0 mg / mL of FOR46 drug substance; 20 mM L-histidine buffer; 8.0% (w / v) sucrose; and 0.01% (w / v) polysorbate 80, pH 6.0. The formulation was designed to provide adequate stability (prevention of antibody denaturation and agglutination), buffering, and cryoprotection for storage at -20°C. After storage at 5°C for one month, the formulation retained >90% binding affinity and cell-based activity; >90% monomer content; <15 μg / mL residual MMAE; and substantially no visible particles. [Example] [4] [Dose escalation study] [-] [use] [FOR46] [Treatment of metastatic castration-resistant prostate cancer]

[0171] A dose-escalation clinical trial was conducted to determine the maximum resistance level (or maximum test dose) of FOR46 in human individuals with metastatic castration-resistant prostate cancer (mCRPC), including treatment-associated small cell / neuroendocrine prostate cancer (tSCNC). Eligible patients had developed one or more androgen signaling inhibitors, maintained castration-resistant testosterone levels (< 50 ng / dL), and demonstrated organ function defined as follows: heme (Hgb) > 8 g / dL, absolute neutrophil count (ANC) > 1500 / μL; platelet count (Plt) > 100kJ; aspartate transaminase:alanine transaminase ratio (ALT / AST) < 2.5 × upper limit of normal (ULN); bilirubin (Bili) < 1.5 mg / dL; and creatinine < 1.5 × ULN. Prior chemotherapy for mCRPC was not permitted. Eligible patients received or were expected to receive FOR 46 via IV infusion every 21 days. Thirty-three individuals were enrolled at 10 doses ranging from 0.1 to 3.0 mg / kg. The median age was 66 (range 42–81); the median baseline PSA was 41 (range 0.2–1627); and seven individuals had visceral metastases. Patient demographics are presented in Table 7. [surface] [7]. Patient demographics in prostate cancer dose escalation trials [feature] [(N = 33)] [Current Data] Median age (range) 67.5 (42-81) gender 33 men Race – White / Asian / Other / Black / American Indian 26 / 1 / 1 / 4 / 1 The median (range) of the previous scheme. 3 (2-8) Disease development type at the start of the study PSA Nodules only (no bone disease) Bone (± nodular diseases) Diseases of internal organs (lungs, liver, adrenal glands, CNS) ± other parts 13 3 16 4 Number of patients with visceral diseases 7

[0172] Accelerated titration was used, followed by a 3+3 dose escalation design. In individuals with high body mass index (BMI) experiencing overdose toxicity (neutropenia and fatigue), the dosage was changed from actual weight to adjusted weight. Secondary prevention of G-CSF was for individuals experiencing ≥ grade 3 neutropenia during a previous treatment cycle. Treatment was continued if the investigator determined potential clinical benefit in the absence of overdose toxicity. A 50% reduction in serum prostate-specific antigen (PSA) levels provided preliminary objective evidence of response to treatment.

[0173] The 33 individuals were divided into 10 groups receiving different doses. Group and patient status are summarized in... [surface] [8] The reduction in PSA and tumor burden is summarized in [surface] [9] In this study, at doses of 1.2 mg / kg or higher (n = 24), 9 individuals (38%) showed a 50% reduction in PSA levels (PSA50 response), and 15 individuals (63%) showed no PSA reduction. Among the 8 individuals with measurable disease, as measured by RECIST criteria, 3 reported objective partial responses (PR), and 6 had stable disease lasting 9 to 39 weeks. Eisenhauer et al., New response evaluation criteria in solid tumors: Revised RECIST guideline (1.1 edition), European Journal of Cancer 45 (2009) 228-249. The median number of treatment cycles was 6 (range 1-28), and 11 were ongoing.

[0174] The PSA levels and RECIST results for groups 4-10 after each infusion cycle are presented in the table below. [surface] [10-16]. The outcomes for all patients are summarized in... [picture] [6] in.

[0175] Patient 12 showed the greatest substantial reduction in tumor burden. A CT scan after three cycles of treatment with 2.7 mg / kg showed complete shrinkage of the largest tumor. [picture] [5A]. The sum of the maximum diameters of the target lesions (SLD, including pulmonary nodules and perirectal soft tissue mass) decreased from 5.7 cm at baseline to 2.0 cm after cycle 6 (65% reduction). At the same time, serum PSA decreased by 71%, and non-target lesions, including RP nodules, also decreased. [picture] [5B].

[0176] Neutropenia is diagnosed by measuring, for example, [surface]

[17] Absolute neutrophil count (ANC) analysis is shown. Grade 2 or higher neutropenia was observed in 12 of the 16 patients treated with at least 1.8 mg / kg FOR46. [surface] [8.] Individual group, patient ID, and current status in the prostate cancer dose escalation trial. EOT: End of treatment; C: Course of disease; D: Day; * Dose increased to 2.1 mg / kg. [Patient ID] [state] [4th] [Group] [(1.2 mg / kg)] 003-04-008 EOT due to disease exacerbation following COVID-19 treatment pause (C9) 004-04-009* C26D1 in progress 001-04-010 EOT due to disease exacerbation (C3) [5th] [Group] [(1.8 mg / kg)] 004-05-011 EOT due to disease exacerbation (C13) 001-05-012 EOT due to disease exacerbation (C3) 004-05-013 EOT due to exacerbated neuropathy (C11) 004-05-014* EOT due to disease exacerbation (C15) 002-05-015 EOT due to disease exacerbation (C5) 004-05-016 EOT due to disease exacerbation (C9) 004-05-017 EOT due to exacerbated neuropathy (C10) [6th] [Group] [(2.4 mg / kg)] 001-06-018 C17D1 in progress 004-06-019 EOT due to disease exacerbation (C6) 004-06-020 Pt decreases before C2D1 - Unable to assess reaction [No. 7] [Group] [(2.1 mg / kg)] 004-07-021 EOT due to disease exacerbation (C3) 003-07-022 EOT due to disease exacerbation (C3) 003-07-023 EOT patient withdrawal (C1) - axillary adenosis; RUE swelling [8th] [Group] [(2.4 mg / kg)] [w / AJBW)] 001-08-024 EOT due to disease exacerbation (C9) 004-08-025 EOT due to disease exacerbation (C10) 001-08-026 No dose reduction (165 mg) was observed during the ongoing C10D1-C10 treatment. 001-08-027 No dose reduction was observed during the ongoing C9D1-C9 treatment (191 mg). [9th] [Group] [(2.7 mg / kg)] [AJBW] 001-09-028 No dose reduction (183 mg) was observed during the ongoing C7D1-C7 treatment. 005-09-029 During the C5D1-C2 process, the dose was reduced to 2.4 mg / kg, and during C3, it was reduced to 1.8 mg / kg. 004-09-030 No dose reduction was observed during the ongoing C6D1-C6 treatment (173.9 mg). [No. 10] [Group] [(3.0 mg / kg)] [AJBW] 003-10-031 The C4D1-C2 dose was reduced to 2.4 mg / kg; the C3 dose was reduced to 2.1 mg / kg. C4 administration was delayed due to colitis, and the dose was reduced to 1.8 mg / kg. 004-10-032 Death due to disease progression (C1) 001-10-033 C3D1 - Dosage reduced to 2.7 mg / kg [Dosage Increase] 003-09-101 C1D1 [surface] [9]. Overview of response to FOR46 in prostate cancer dose escalation trials. PR: Partial Response [Patient ID] [dose] [(mg / kg)] [% PSA] [change] [PSA] [change] [RECIST] [#] [cycle] 003-04-008 1.2 +37 9 004-04-009 1.2 -94 ≥50% 26+ 001-04-010 1.2 -51 ≥50% 3 004-05-011 1.8 +75 14 001-05-012 1.8 +34 3 004-05-013 1.8 -56 ≥50% 13 004-05-014 1.8 -31 red 14 002-05-015 1.8 -14 red 2 004-05-016 1.8 -50 ≥50% 9 004-05-017 1.8 -79 ≥50% 9 001-06-018 2.4 -51 ≥50% 18+ 004-06-019 2.4 +76 6 004-07-021 2.1 +55 3 003-07-022 2.1 -34 red 5 001-08-024 2.4 AJBW -12 red 9 004-08.025 2.4 AJBW +27 10 001-08-026 2.4 AJBW -3 red 4 001-08-027 2.4 AJBW +20 9 001-09-028 2.7 AJBW -71 ≥50% PR 10+ 004-09-030 2.7 AJBW -79 ≥50% 7+ 003-10-031 3.0 AJBW 34 PR 4+ 001-10-033 3.0 AJBW -74 ≥50% PR 4+ [] [, , ] [surface]

[10] . Serum PSA (mg / ml) levels and tumor size (RECIST) in group 4 metastatic castration-resistant prostate cancer patients treated with 1.2 mg / kg FOR46. SCR: screening; C: disease duration; D: day; N / N: incomplete response, non-progressive disease; SD: stable disease; PD: progressive disease; * The dose was increased to 2.1 mg / kg AJBW at C17. [patient] [ID] [SCR] [C1] [C2] [C3] [C4] [C7] [C10] [C14] [C18] [C22] [C26] 003-04-008 RECIST 399 98 mm 352.9 419.3 414.2 484.0 SD -6.1% 92 mm 507 SD -9.8% 88 mm EOT PD -- -- -- [] 004-04-009* RECIST 78.7 NM 9.4 2.4 0.58 0.66 N / N 1.17 N / N 4.13 N / N 10.4 N / N 14.2 N / N 17.37 N / N 19.33 N / N 001-04-010 RECIST 1603 18 mm 1626 >149 794.6 1502 PD -- -- -- -- [surface] [11 (] [No.] [1] [part] [].] Serum PSA (mg / ml) levels and tumor size (RECIST) in group 5 patients with metastatic castration-resistant prostate cancer treated with 1.8 mg / kg FOR46. SCR: Screening; C: Disease duration; D: Day; SD: Stable disease; PD: Progressive disease; *004-05-014 Discontinued after cycle 15 due to peripheral neuropathy and fatigue / asthenia; AE: Adverse events. [patient] [ID] [SCR] [C1] [C2] [C3] [C4] [C5] [C6] [C7] [C8] [C9] [C10] 004-05-011 RECIST 45.6 43 55.0 78.8 95.0 96.1 SD 47 98.9 126.7 146.4 SD 51 163.5 171.5 221 SD 52 001-05-012 RECIST 382 134 545.5 623.6 731.8 1027 PD 174 -- -- -- -- -- -- 004-05-013 RECIST 57.7 63 57.5 28.4 25.4 26.6 SD -14.2% 54 31.6 42.2 59.0 SD -9.5% 57 66.2 55.0 53.4 SD -6.77% 59 004-05-014 RECIST 129 40 152.7 198.4 141.1 150.1 SD -7.5% 37 109 106.2 111 SD -17.5% 33 113.9 104.9 127.7 SD -14.2% 35 002-05-015 RECIST 685 NM 884 758 991 1179 N / N 1136 1147 -- -- -- -- 004-05-016 RECIST 91.2 NM 71.5 81.7 54.6 49.1 N / N 35.7 48 54.8 N / N 54.7 77.5 99.1 PD new lesions 004-05-017 RECIST 1.49 NM 1.44 0.8 0.66 1.0 N / N 0.4 0.3 0.3 N / N 0.4 0.73 1.8 EOT N / N [surface] [11 (] [continue] [).] Serum PSA (mg / ml) levels and tumor size (RECIST) in group 5 patients with metastatic castration-resistant prostate cancer treated with 1.8 mg / kg FOR46. C: Disease duration; D: Day; SD: Stable disease; PD: Progressive disease; *004-05-014 Discontinued after cycle 15 due to peripheral neuropathy and fatigue / asthenia; AE: Adverse events. [patient] [ID] [C11] [C12] [C13] [C14] [C15] 004-05-011 RECIST 235 254 250.8 312.6 53 -- 001-05-012 RECIST -- -- -- -- -- 004-05-013 RECIST 60.6 48.27 53.79 64.27 PD -- 004-05-014 RECIST 135.5 173.3 212.5 256.9 SD -17.5% 33 297.3 Stopped due to AE* 002-05-015 RECIST 004-05-016 RECIST 004-05-017 RECIST [surface] [12.] Serum PSA (mg / ml) levels and tumor size (RECIST) in group 6 metastatic castration-resistant prostate cancer patients treated with 2.4 mg / kg FOR46. SCR: screening; C: disease duration; D: day; SD: stable disease; N / N: incomplete response, non-progressive disease; NE: not evaluable. [patient] [ID] [SCR] [C1] [C2] [C3] [C4] [C5] [C6] [C7] [C8] 001-06-018* RECIST 47.6 NM 16.4 14.5 8.1 9.8 N / N 16.5 19.2 22.3 N / N 28.9 004-06-019* RECIST 3.53 NM 6.93 15.3 12.2 15.3 N / N 18.7 25.3 43.6 -- 004-06-020 RECIST (mm) 7.5 179 5.6 NE -- -- -- -- -- -- -- [patient] [ID] [C9] [C10] [C11] [C12] [C13] [C14] [C15] [C16] [C17] 001-06-018* RECIST 25.2 21.2 N / N 17.8 14.2 17.5 16.1 N / N 17.8 21.7 19.1 004-06-019* RECIST -- -- -- -- -- -- -- -- 004-06-020 RECIST (mm) -- -- -- -- -- -- -- -- [surface] [13.] Serum PSA (mg / ml) levels and tumor size (RECIST) in group 7 patients with metastatic castration-resistant prostate cancer treated with 2.1 mg / kg FOR46. SCR: screening; C: disease duration; PD: progressive disease; NE: not evaluable; *dose reduced from C2 to 1.8 mg / kg actual body weight; #dosage based on AJBW. [patient] [ID] [SCR] [C1] [C2] [C3] [C4] [C5] 004-07-021 RECIST 3.88 104 4.2 5.5 6.5 -- 113 -- 003-07-022 RECIST (mm) 24.7 NM 34.1 31.4 27.7 20.7 PD 22.7 003-07-023# RECIST (mm) 119.5 138 487 NE -- -- -- [surface] [14.] Serum PSA (mg / ml) levels and tumor size (RECIST) in group 8 patients with metastatic castration-resistant prostate cancer treated with 2.4 mg / kg (adjusted body weight) FOR46. SCR: screening; C: disease duration; NM: unmeasurable; SD: stable disease; N / N: incomplete response, non-progressive disease. [patient] [ID] [SCR] [C1] [C2] [C3] [C4] [C5] [C6] [C7] [C8] [C9] [C10] [C11] 001-08-024 RECIST 5.8 52 mm 5.8 5.1 5.3 5.9 SD 50 mm 7.1 9.5 13.5 SD 55 mm 14.2 15.2 21.75 PD 67 mm [] 004-08-025 RECIST 164 61 mm 130 186 181 222 SD 63 mm 167 217 206 SD 229.8 250.5 301 357 EOT 001-08-026 RECIST 0.68 NM 0.62 0.60 0.64 0.75 N / N 0.98 1.4 0.7 N / N 2.1 1.6 1.921 N / N 001-08-027 RECIST 82.6 NM 79.5 97.8 95.4 117 N / N 135 160 153.8 N / N 170.7 190.8 [surface] [15.] Serum PSA (mg / ml) levels and tumor size (RECIST) in group 9 patients with metastatic castration-resistant prostate cancer treated with 2.7 mg / kg (adjusted body weight) FOR46. SCR: screening; C: disease duration; PR: partial response; NM: unmeasurable; N / N: incomplete response, non-progressive disease. [patient] [ID] [SCR] [C1] [C2] [C3] [C4] [C5] [C6] [C7] [C8] 001-09-028 RECIST 22.4 57 mm 24.9 10.9 10.9 15.5 25 mm PR 31.0 33.97 54.5 20 mm PR 005-09-029 RECIST 0.2 NM 0.20 0.21 0.20 0.20 N / N 004-09-030 RECIST 134 NM 162 96 52 34.2 N / N 34.7 30.81 [surface] [16.] Serum PSA (mg / ml) levels and tumor size (RECIST) in group 10 patients with metastatic castration-resistant prostate cancer treated with 3.0 mg / kg (adjusted weight) FOR46. SCR: screening; C: disease course; PR: partial response; D / C: discontinued care; *C2 dose reduced to 2.4 mg / kg (adjusted weight); #C2 dose reduced to 2.7 mg / kg (adjusted weight). [patient] [ID] [SCR] [C1] [C2] [C3] [C4] 003-10-031* RECIST 5.80 24 mm 2.58 1.7 1.78 2.09 13 mm 004-10-032 RECIST 51.32 65.99 D / C Anning House 001-10-033# RECIST 188.9 16 cm 221.6 65.4 57.3 50.8 8.5 cm PR [surface]

[17] [.] Absolute neutrophil count (×10⁹ / L) in patients with metastatic castration-resistant prostate cancer treated with FOR46. Neutropenia (Grade 2; Grade 3; Grade 4); * G-CSF; ** Adjusted weight; *** Adjusted weight – incomplete dose due to infusion reaction; D / C: discontinued care. [patient] [ID] [SCR] [Wt] [AJBW] [Ht] [BMI] [dose] [C1D1] [C1D8] [or] [9] [C1D15] [C2D1] [C2D8] [C2D15] 001-06-018 6.54 114.7 185 33.5 275 2.83 ^^^^0.19 ^1.04 5.63 3.46 ^^1.17 004-06-019 2.34 126.8 182.9 37.9 304.3 2.47 ^^^^0.1* -- 2.37 4.6 4.1 004-06-020 7.00 123.6 177.8 39.1 296.6 4.86 ^^^^0.21* -- -- -- -- 004-07-021 4.09 76.6 179.1 23.9 160.9 3.52 2.72 2.98 4.8 1.63 ^^1.47 003-07-022 6.2 68 166 24.7 141.5 4.6 2.3 ^^^0.7 6.4 13.7 7.0 003-07-023 8.0 112.4 180.3 34.6 189.2** 7.8 3.1 2.2 -- -- -- 001-08-024 2.96 112.6 176.5 36.1 213*** 2.60 2.47 3.31 1.98 2.61 1.93 004-08-025 4.43 71.7 169 25.1 172.6** 4.70 ^^1.42 ^^^^0.31 4.37 8.18 7.40 001-08-026 4.12 73.9 169.5 25.7 165** 2.28 ^^1.11 ^^1.31 3.81 ^^1.16 ^^^0.76 001-08-027 4.67 94.5 172.7 31.9 191** 3.60 2.39 3.45 4.88 3.40 3.95 001-09-028 2.53 75 68 167 26.9 183 2.50 ^^^0.65 ^^^0.70 2.80 5.50 4.29 005-09-029 10.68 90.9 80 178 28.7 217.1 4.98 ^^^^0.39 3.23 10.90 1.95 ^^^0.8 004-09-030 4.02 64.1 - 172.7 21.5 173.9 5.10 1.89 ^^1.38 2.90 ^^1.33 ^^^^0.09 003-10-031 2.4 80.8 73 172.7 27.1 220.2 2.2 ^^^^0.1 0.4 1.8 5.4 6.6 004-10-032 4.8 95.6 87 186.5 27.5 261 4.14 ^^^^0.23 0.1 (Day 11) D / C Anning House 001-10-033 2.16 85.2 191.2 23.3 255 3.53 2.02 .895 (Day 6) 1.45 1.30 2.70 3.15 003-09-101 2.6 83 164.5 30.7 186.6 3.1

[0177] Dose-limiting toxicity was grade 4 neutropenia in 3 out of 3 individuals with high body mass index (BMI) who received 2.4 mg / kg based on actual body weight, and in 2 out of 3 individuals who received 3.0 mg / kg based on adjusted body weight. The maximum tolerated dose (MTD) based on adjusted body weight (AJBW) was 2.7 mg / kg. The most common associated adverse events were grade 4 neutropenia in 11 out of 33 individuals (33%), grade 3 neutropenia in 6 individuals (18%), infusion-related reactions (IRR) in 14 individuals (42%), and grade 3 IRR in 1 individual. Neuropathy of any grade occurred in 7 individuals (21%), and grade 3 neuropathy in 1 individual (3%). The most common adverse events were shown in [surface]

[18] [surface] [18.] [use] [FOR46] [Adverse events observed in at least two individuals with prostate cancer] [.] [Adverse Event] [Number of patients n (%) n = 35] [Lowest Patient Level] [Any level] [3] [4] Infusion-related reactions 14 (40) 1 (3) - Neutropenia 12 (34) 3 (9) 5 (14) Decreased neutrophil count 10 (29) 3 (9) 6 (17) Decreased white blood cell count 8 (23) 3 (9) 1 (3) fatigue 7 (20) 1 (3) - Peripheral neuropathy 7 (20) 1 (3) - diarrhea 6 (17) 1 (3) - anemia 5 (14) 1 (3) - Decreased lymphocyte count 5 (14) 1 (3) 1 (3) nausea 5 (14) - - hair loss 5 (14) - - hypokalemia 4 (11) - - ALT increases 3 (9) - - constipate 3 (9) - - Loss of appetite 3 (9) - - hypomagnesemia 3 (9) - - Leukopenia 2 (6) - 1 (3) Lymphopenia 2 (6) - - AST increases 2 (6) - - Increased liver enzymes 2 (6) - - Fear of cold 2 (6) - - fever 2 (6) - - Headache 2 (6) - - hyponatremia 2 (6) 1 (3) - hypophosphatemia 2 (6) - - Difficulty breathing 2 (6) - -

[0178] A dose-expansion study has been initiated in individuals with prostate cancer. CD46 expression was determined at enrollment using immunofluorescence microscopy. Three patients with intermediate or strong positive CD46 expression have been enrolled. A fourth individual with negative CD46 expression has not been enrolled.

[0179] This example demonstrates acceptable toxicity with weight-adjusted dosing of FOR46 and provides encouraging preliminary evidence of efficacy in individuals with androgen signaling inhibitor-resistant mCRPC. FOR46 is currently being evaluated in two expanded mCRPC cohorts: adenocarcinoma and t-SCNC. [Example] [5.] [Dose escalation study] [] [–] [] [use] [FOR46] [Treatment of relapsed or refractory multiple myeloma]

[0180] A dose-escalation clinical trial was conducted to treat human individuals with relapsed or refractory multiple myeloma using the FOR46 drug product described in Example 2. To be eligible, patients' prior therapy must have included a proteasome inhibitor, an immunomodulatory amide (ImiD), and CD38-directed therapy. Eligible patients also had the following organ function indicators: heme ≥ 8 g / dL, ANC ≥ 1500 / µL; platelets ≥ 100kJ; ALT / AST ≤ 2.5 × upper limit of normal (ULN); bilirubin ≤ 1.5 mg / dL; and creatinine ≤ 1.5 × ULN. FOR46 was administered every three weeks via intravenous infusion using an infusion-related reaction prevention protocol over 30–60 minutes.

[0181] The initial regimen had a maximum dose of 2.4 mg / kg actual weight. Without using adjusted body weight to define the MTD, the escalation was maintained in a pending regimen revision to allow for higher doses.

[0182] A dose expansion clinical trial was also conducted on 10 patients who received FOR46 at an adjusted weight of 2.4 mg / kg. The eligibility criteria for the dose expansion trial were the same as those for the dose escalation trial, except that ANC ≥ 1000 / µL and platelets ≥ 75kJ.

[0183] For the dose escalation trials, fifteen individuals were enrolled at six predetermined dose levels ranging from 0.1 to 2.4 mg / kg. One patient received doses of 0.1, 0.3, and 0.6 mg / kg, three received 1.2 and 1.8 mg / kg, and six received 2.4 mg / kg. The median age was 68 (range 33–79), and four were female. An increase of 1q occurred in nine patients, was not observed in five, and was unknown in one. The median preceding line of treatment was 6 (range 3–17). Dosing administration in the dose escalation and dose expansion trials is shown in [data missing]. [surface]

[19] In. Patient characteristics are shown in [surface]

[20] [and]

[21] [surface] [19.] Dosing of FOR46 in dose escalation and dose expansion trials for relapsed or refractory multiple myeloma. [Dose Level] [(mg / kg q 3] [week] [)] [N (25)] 0.1 1 0.3 1 0.6 1 1.2 3 1.8 3 2.4 (Incremental – Actual / AJBW Dosing) 6 (3 / 3) 2.4 (Extended – AJBW) 10 [surface] [20.] Demographic data of individuals in dose escalation and dose expansion trials of FOR46 for relapsed or refractory multiple myeloma. [feature] [Incrementing] [(n = 15)] [and expansion] [(n = 10)] Median age (range) 67 (33-79) Gender F / M 7 / 18 Race – White / Black / Hispanic / Unknown 19 / 2 / 1 / 2 Myeloma light chain κ LC 18 (only 2 light chains) λ LC 6 Immunoglobulins IgA 6 IgG 15 IgM 1 [surface] [21.] Prior treatment in individuals in dose escalation and dose expansion trials of FOR46 for relapsed or refractory multiple myeloma. [Previous treatment,] [n (%) [Incrementing] [(n=15)] [and expansion] [(n = 10)] Median (range) 8 (3-19) Receive ≥ 5 lines of treatment 21 (84) Proteasome inhibitors, accepted / refractory 25 (100) / 21 (84) IMiD, Acceptable / Refractory 25 (100) / 22 (88) Pomalidomide, accepted / refractory 20 (80) / 18 (72) Anti-CD38 therapy, for patients who have received / are refractory treatment 25 (100) / 23 (92) Carfilzomib, accepted / refractory 23 (92) / 23 (92)

[0184] Accelerated titration was performed in dose escalation studies, followed by a 3+3 dose escalation design. FOR46 was administered intravenously over 30–60 minutes on day 1 of a 21-day cycle at the dose specified in the protocol. In individuals with high body mass index (BMI) who experienced overdose toxicity (neutropenia and fatigue), the dosage was changed from actual body weight (AW) to adjusted body weight (AJBW). G-CSF secondary prevention was administered to individuals who experienced ≥ grade 3 neutropenia in a previous treatment cycle.

[0185] Safety was assessed using the Common Terminology Criteria for Adverse Events (CTCAE) v5.0. Dexamethasone was used only for infusion reaction prophylaxis. CD46 antigen density was determined on the patient's MM cells by flow cytometry. Treatment efficacy was monitored by measuring the levels of immunoglobulins (M-proteins) in serum or urine. Immunoglobulins include IgA, λ light chain (λ), κ light chain (κ), and M-spike protein.

[0186] The only dose-limiting toxicity was grade 4 neutropenia in one high-BMI patient treated with AW. This was the only dose-limiting toxicity in six patients treated with a mixture of AW (n=3) and ABW (n=3) at 2.4 mg / kg. One of the three patients treated with 2.4 mg / kg AJBW experienced non-dose-limiting grade 4 neutropenia. In three patients (20%), the most common associated adverse event was grade 4 neutropenia. One patient (6.7%) had grade 4 thrombocytopenia, and one patient (6.7%) had grade 3 elevated AST, neutropenia, anemia, nausea, and peripheral neuropathy (PN). Adverse events were shown in [surface] [twenty two] [middle]. [surface] [22.] Adverse events in individuals during dose escalation and dose expansion trials of FOR46 for relapsed or refractory multiple myeloma. [Adverse Reactions] [Number of patients] [n (%) n = 25] [Any level] [3] [class] [4] [class] Neutropenia 6 (24) 2 (8%) 1 (4%) anemia 5 (20) 4 (16%) - AST increase 4 (16) 2 (8%) - Decreased neutrophil count 4 (16) 2 (8%) 2 (8%) Decreased platelet count 3 (12) 1 (4%) 1 (4%) weight loss 3 (12) - - constipate 3 (12) - - nausea 3 (12) 1 (4%) - fatigue 3 (12) - - Decreased white blood cell count 2 (8) 2 (8%) - diarrhea 2 (8) - - Vomit 2 (8) - - fever 2 (8) - - Joint pain 2 (8) - - Headache 2 (8) - - Peripheral neuropathy 2 (8) - - hair loss 2 (8) - -

[0187] In the initial assessment, all patients receiving FOR46 at doses less than 1.8 mg / kg (i.e., 0.1 mg / kg, 0.3 mg / kg, 0.6 mg / kg, and 1.2 mg / kg) discontinued treatment due to disease progression. Treatment has been initiated for patients in the 1.8 mg / kg group. Patient 8 showed a response to FOR46 treatment, with reductions in serum IgG, serum κ light chain, serum λ light chain, and urinary M-spike protein. This response provides preliminary evidence of antitumor activity at the 1.8 mg / kg dose.

[0188] Four patients responded to FOR46, with partial regression (PR) per IMWG criterion. (BGM Durie et al. International uniform response criteria for multiple myeloma. Leukemia (2006) 1-7.) See also [surface] [twenty three]. [surface] [23.] Multiple myeloma patients who respond to FOR46 [patient] [ID] [1q] [Increase] [C1D1 / ] [#] [cycle] [Ig] [serum] [FLC (mg / dL)] [serum] [M-] [Spike protein] [(g / dL)] [Urine] [M-] [Spike protein] [(g / 24)] [Hour] [)] [optimal] [IMWG] [reaction] [Types of Myeloma] [Dose Level] 006-05-008 Neg 2 / 5 / 20 4002 573 2.84 20.35 [PR] IgG κ 1.8 mg / kg 7 cycles 1193 12.4 0.44 IFE+; M-spike protein absent 001-06-012 Pos 6 / 29 / 20 1440 21.2 1.4 - [PR] IgA κ 2.4 mg / kg ABW 12 353 10.7 0.3 - 003-06-014 Pos 10 / 21 / 20 187 (wnl) 68.1 det 540 [PR] IgA κ 9 168 34.8 det 95 001-06-102 Pos 2 / 3 / 21 3520 131.6 2.5 - [PR] IgG λ 7 2300 56.7 1.1 -

[0189] Of the six evaluable patients in the 1.8 and 2.4 mg / kg dose escalation groups, three achieved partial responses (PRs) lasting 21, 30, and 15 weeks, respectively. One patient did not achieve a 1q21 increase in PR. In the dose escalation group, three of the ten patients were not evaluable. Of the seven evaluable patients, one achieved a PR lasting 18 weeks, but discontinued a partial response due to an adverse event of peripheral neuropathy. Two patients had persistent stable disease in the three and six-cycle groups. Four patients achieved optimal responses in patients with progressive disease.

[0190] Patient 006-05-008 was treated with 1.8 mg / kg FOR46. This patient was a 62-year-old white male who was diagnosed with IgG κ MM in July 2009. The patient had a negative increase in 1q and had previously been treated with: (1) daratumumab, pomalidomide and dexamethasone; (2) pomalidomide and dexamethasone; (3) lenalidomide; (4) lenalidomide and bortezomib; and (5) carfilzomib and pomalidomide. IgG, κ light chain and serum M-spike protein results were shown in... [picture] [7A] in.

[0191] Patient 001-06-012 was treated with 2.4 mg / kg FOR46. This patient was a 70-year-old white male who was diagnosed with IgA κ MM in January 2013; the patient had a positive increase in 1q and had previously been treated with: (1) cyclophosphamide, bortezomib and dexamethasone; (2) lenalidomide, bortezomib and dexamethasone; (3) carfilzomib, cyclophosphamide and dexamethasone; and (4) daratumumab, pomalidomide and dexamethasone. The results of IgA, κ light chain and serum M-spike protein are shown in Figure 1. [7B] in.

[0192] Patient 003-06-014 was treated with 2.4 mg / kg (AJBW) for 46. This patient was a 56-year-old male who was diagnosed with IgA κ myeloma in December 2015. The patient had positive 1q21 elevation and had previously been treated with: (1) cyclophosphamide, bortezomib, and dexamethasone; (2) carfilzomib, lenalidomide, dexamethasone, melphalan, and ASCT, maintained with ixazomib; (3) carfilzomib, daratumumab, and dexamethasone; and (4) a CAR-T clinical trial. IgA, κ light chain, and urinary M-spike protein results were shown in... [picture] [7C] in.

[0193] Results for all patients in the dose escalation trial are presented in Table 24. Results for all patients in the dose expansion trial are presented in Table 25. Results from both trials are summarized in Figure 8.

[0194] In summary, FOR46 administered with body weight adjustment showed acceptable toxicity. Encouraging evidence of efficacy was found in triple-refractory multiple myeloma. A dose escalation trial increased the dose to 2.7 mg / kg based on body weight. Table 24. Biomarker results in patients with refractory multiple myeloma treated with FOR46 in a dose escalation trial. Dosage units are mg / kg; C: duration of disease; D: day; EOT: end of treatment; K: κ light chain; λ: λ light chain. Unless otherwise indicated, M spike protein levels were measured in serum. [patient] [(] [dose] [)] [Analytical material] [Reference Range] [Screening] [C1] [D1] [C2] [D1] [C3] [D1] [C4] [D1] [C5] [D1] [C6] [D1] [C7] [D1] [C8] [D1] [C9] [D1] [C10] [C11] [C12] [EOT] [or status] [1] [(0.1)] IgA 672-1760 mg / dL 2230 EOT 3750 K 5.7-26.3 mg / L 1718.9 1772.4 EOT 3330.2 M-spike protein Undetectable 1.5 2.1 2.5 [] [2] [(0.3)] K 3.3 - 19.4 mg / L 179.9 201.4 EOT 648.7 M-spike protein Undetectable 0 [3] [(0.6)] IgG 672-1760 mg / dL 3570 3560 3710 EOT 3560 K 3.3 - 19.4 mg / L 9.3 11.3 9.5 EOT 12.6 M-spike protein Undetectable 2.8 2.7 EOT 2.8 [] [4] [(1.2)] IgG 672-1760 mg / dL 1160 1230 1250 1380 1570 1690 EOT 1980 K 3.3 - 19.4 mg / L 107.9 137.3 185.8 203.8 293.5 318.3 EOT 452.4 M-spike protein 0 g / dL 0.9 1.0 1.2 1.2 1.4 1.5 [] [5] [(1.2)] IgG 700-1600 mg / dL 1551 1539 1762 - - - EOT 2012 λ 5.7 - 26.3 mg / L 281.8 331.7 415.1 - - - 427.7 M-spike protein 0 g / dL 1.11 1.07 1.25 - - - EOT 1.43 [] [6] [(1.2)] K 3.3 - 19.4 mg / L 3780.3 7915.0 3874.6 C2D8 8919.2 EOT 14833.4 M-spike protein 0 gm / dL 0.24 0.45 0.47 0.69 EOT 0.86 M-spike protein (urine) 0 gm / dL 268.1 [] [7] [(1.8)] [1q21 pos] IgG 635-1741 mg / dL 2910 2990 3288 3596 K 0.33-19.4 mg / dL 258.00 290.0 398.0 EOT 1183 EOT 1183 M-spike protein Unable to detect g / dL 2.30 2.27 2.35 [] [8] [(1.8)] [1q21 neg] IgG 610-1616 mg / dL 3049 4002 1899 1233 1193 1231 1373 1573 1594 1681 K 0.33-19.4 mg / dL 296.2 573.0 14.1 16.3 12.4 12.9 14.2 21.1 36.6 79.2 M-spike protein Unable to detect g / dL 1.86 2.84 0.87 0.44 0.50 0.56 0.60 0.72 1.01 0.96 M-spike protein (urine, 24 hours) Unable to detect g / dL 1.1 IFE+; M-spike protein absent IMWG reaction PR PR PD EOT [] [9] [(1.8)] [1q21 neg] IgA 66-433 mg / dL 945 1309 1328 1285 1450 1436 1566 1510 1556 K 0.33-1.94 mg / dL 3.33 3.93 4.66 5.11 5.17 5.62 6.50 7.67 8.65 M-spike protein 0 g / dL Det Det Det Det Det Det Det Det M-spike protein (urine, 24 hours) N / D Det Det N / D IMWG reaction SD SD 10 (2.4) [1q21 neg] K 1037.5 1074 1485 M-spike protein 0 g / dL 0.06 0 IMWG reaction 11 (2.4) [1q21 pos] IgG 610-1616 mg / dL 1466 1585 K 0.33-19.4 mg / dL 20.3 26.5 M-spike protein 0 g / dL 0.63 0.68 M-spike protein (urine, 24 hours) 0 g / dL 114.7 IMWG reaction 12 (2.4) [1q21 pos] IgA 1510 1440 1120 948 799 653 698 634 492 417 394 353 391 K 3.3 - 19.4 mg / L 18.3 21.2 18.1 18.0 16.3 14.6 12.3 12.2 10.7 12.5 10.9 13.6 12.6 Serum M-spike protein 0 g / dL 1.4 1.0 0.8 0.6 0.5 0.5 0.4 0.4 0.3 0.3 0.3 0.3 IMWG reaction PR PR PR termination 13 (2.4) [1q21 pos] IgG 672-1760 mg / dL 2010 K 3.3 - 19.4 mg / L 135.2 139.0 Serum M-spike protein 0 g / dL 2.5 IMWG reaction 14 (2.4) [1q21 pos] IgG 635-1741 mg / dL 345 339 K 0.33-1.94 mg / L 28.72 68.11 48.16 42.16 36.18 34.78 35.57 M-spike protein (urine, 24 hours) 0 mg / day 540 144 202 95 IMWG reaction PR MR PR 15 (2.4) [1q21 neg] IgG 635 - 1741 mg / dL 345 339 Κ .33 – 1.94 mg / L 2.61 3.67 <00​​​​​​​​​​​​​Serum M-spike protein 0 g / dL det Incomplete IMWG reaction NE [surface] [25.] Biomarker results in patients with refractory multiple myeloma treated with FOR46 at 2.4 mg / kg (adjusted body weight) in a dose expansion trial. C: duration of disease; D: day; EOT: end of treatment; K: κ light chain; λ: λ light chain; PR: partial response; PD: progressive disease; D / C: discontinued care. Unless otherwise indicated, M spike protein levels were measured in serum. [patient] [ID] [Test Name] [Reference Range] [Screening] [C1D1] [C1D15] [C2D1] [C3D1] [C4D1] [C5D1] [C6D1] 005-06-101 1q increases pos IgG 610 - 1616 mg / dL 3079 3361 3301 4029 Serum λ light chain 5.7 - 26.3 mg / dL 129 142.7 191.9 303.6 Serum M-spike protein 0 g / dL 2.14 2.22 2.6 3.21 IMWG reaction PD 001-06-102 1q increases pos (87%) IgG 672-1760 mg / dL 3440 3520 2300 1770 1310 1220 1530 Serum λ light chain 5.7-26.3 mg / dL 131.6 156 56.7 31.6 59.4 97.7 128 Serum M-spike protein 0 g / dL 2.4 2.5 1.6 1.5 1.1 1.0 Pending IMWG reaction PR 006-06-103 1q Added pos IgG 610-1616 mg / dL 8132 8441 8949 8949 D / C Serum λ light chain 5.7-26.3 mg / L 384.1 328.1 477.4 442.2 Serum M-spike protein 0 g / dL 6.35 6.53 7.98 7.54 PD IMWG reaction 005-06-104 1q increase? IgG 672 – 1760 mg / dL 1872 1706 1357 Serum λ light chain 3.3 – 19.4 mg / dL 1790 838 1113 Serum M-spike protein 0 g / dL 1.33 1.37 1.31 IMWG reaction 001-06-105 1q increases neg IgG 672-1760 mg / dL 7350 7800 7150 Serum λ light chain 3.3 – 19.4 mg / dL 141 106.7 131.9 Serum M-spike protein 0 g / dL 4.7 5.0 5.0 IMWG reaction 006-06-106 1q increases pos 88% of cells have 3 copies IgG 35 – 242 mg / dL 3548 2820 2687 2992 Serum λ light chain 5.7 – 26.3 mg / dL 49.4 46.5 77.7 82.0 Serum M-spike protein (M1+M2) 0 g / dL 1.79 1.43 1.42 1.44 IMWG reaction 001-06-107 1q increases pos IgG 672 – 1760 mg / dL 3870 3900 3940 4990 EOT Serum λ light chain 3.3 – 19.4 mg / dL 121.2 106.6 174.1 272.7 Serum M-spike protein 0 g / dL 3.2 3.5 3.6 IMWG reaction 001-06-108 1q increases pos IgG 672 – 1760 mg / dL 5250 5550 5430 5630 5280 EOT Serum λ light chain 3.3 – 19.4 mg / dL 963.8 1022.5 643.1 1162 1247 Serum M-spike protein 0 g / dL 3.9 3.9 3.7 3.6 3.4 IMWG reaction 001-06-109 1q increases pos IgG 672-1760 mg / dL 3430 5050 Serum λ light chain 3.3 – 19.4 mg / dL 15.9 Serum M-spike protein 0 g / dL 2.9 LDH 125-243 U / L 710 IMWG reaction 002-06-110 1q increases pos IgG 35 – 242 mg / dL 660 611 Serum λ light chain 5.7 – 26.3 mg / dL 2.09 Serum M-spike protein (M1+M2) 0 g / dL IMWG reaction [] [, , ] [Example] [6] [:] [FOR46] [The ingredients]

[0195] The goal of this study was to develop an optimized formulation of FOR46. Thermal stability studies, freeze-thaw stability studies, and stirring studies were conducted during the formulation development process. The stability of the drug product was evaluated using analyses including: approximate appearance, protein concentration, pH, and SEC-HPLC, cIEF, Caliper-SDS_R / NR, and MFI analyses to select the optimal formulation. [Analysis Methods] [] [Appearance]

[0196] The appearance of all samples (including clarity, color, and visible particles) was tested using a YB-2 lightbox against a black and white background. [pH]

[0197] The pH of the samples was measured using a Seven Multi S4.0 pH meter with an Inlab® Micro electrode. The pH meter was calibrated before each use. [Protein Concentration]

[0198] Protein concentrations were determined using a NanoDrop 2000 spectrophotometer with UV280 readings. The extinction coefficient used in all evaluation studies was 1.571 AU*mL*mg⁻¹*cm⁻¹. All measurements were repeated twice using 2.5 µL samples, and the average results were reported. [SEC-HPLC]

[0199] Particle size distribution chromatography was performed at 25°C using an Agilent 1260 Infinity system with a TSKGel G3000SWXL particle size distribution chromatography column (300 × 7.8 mm, 5 gm). The flow rate was set at 1.0 mL / min in an isocratic gradient. For each sample, the mobile phase consisted of 50 mM sodium phosphate buffer and 300 mM NaCl (pH 6.8 ± 0.1). A loading of 100 µg of sample was injected, and the samples were detected using a UV detector at 280 nm. Data were analyzed using Waters Empower. [cIEF]

[0200] cIEF was performed on a ProteinSimple iCE3 device with an FC-coated cIEF cartridge. During the formulation development phase, 50 µg of each sample was mixed with 100 μL of a host mixture, which consisted of pI labels 4.22 / 7.46, Servalyt 2–9, Servalyt 3–5, 1% methylcellulose solution, and 8 M urea solution. After mixing, the samples were incubated at 1500 V for 1 minute and then at 3000 V for 8 minutes. The detection wavelength was set to 280 nm to evaluate the charge variant distribution across different pI ranges. In the forced decomposition study, the pI label in the host mixture was changed to 4.22 / 7.05. [Caliper-SDS_R&NR]

[0201] Pretreatment before testing the samples, such as incubation at 70°C for 10 minutes with sample buffer, SDS, and N-ethylcis-butenediamine (for non-reducing or NR) or dithiothreitol (for reducing or R), is necessary. Subsequently, a minimum loading volume of 42 µL (resulting in a final protein concentration of 0.045 mg / mL) was tested using LabChip GXII Touch at excitation / emission wavelengths of 635 and 700 nm. Final results were analyzed using the commercial software LabChip GX Reviewer. [CE-SDS_R / NR]

[0202] Non-reducing CE-SDS was performed using a Beckman Coulter PA800 Enhanced or PA800 Plus instrument equipped with a photodiode array detector. Samples were diluted to 4 mg / mL with PB-CA and then heated at 60°C for 10 minutes in the presence of 75 µl SDS sample buffer and 5 µl 100 mM NEM for non-reducing CE-SDS. Samples were injected at +5 kV for 15 seconds, followed by separation at +11 kV for 30 minutes. Detection was performed at 220 nm. [DSC] [analyze]

[0203] DSC analysis was performed using a MicroCal™ VP-Capillary DSC system (model AS12-001C) from GE Healthcare. First, protein samples were diluted to 1 mg / mL with formulation buffer before analysis. 300 µL of the protein sample was added to a 96-well plate, along with 300 µL of the corresponding buffer as a reference. Samples were heated from 10°C to 110°C at a heating rate of 200°C / hour in the capillary DSC system. Samples were tested twice, and DSC results (Tm onset and Tm value) were analyzed using Origin 7.0 DSC automated analysis software. [3.] [Excipient Screening] [] [3.1] [Research Objectives]

[0204] This study aimed to evaluate the effects of NaCl, Arg-HCl, sucrose, and trehalose on stabilizing FOR46 in the selected buffer. [3.2] [Research Parameters]

[0205] FOR46 was prepared at a concentration of 10 mg / mL in 20 mM histidine buffer (pH 6.0). As shown in Table 1, each formulation was stabilized by adding 140 mM NaCl, 150 mM Arg-HCl, 8% (w / v) sucrose or trehalose, and those without any stabilizer were set as blanks.

[0206] Each formulation underwent up to five cycles of freeze / thaw stress and thermal stress (40°C and 25°C). The FOR46 stability of each formulation was determined using... [surface] The different analyses given in

[26] are evaluated. [surface] [26.] [Distribution Options] [.] [F#] [Buffer] [pH] [excipient] [F1] 20 mM histidine 6.0 140 mM NaCl [F2] 20 mM histidine 6.0 150 mM Arg-HCl [F3] 20 mM histidine 6.0 8% sucrose [F4] 20 mM histidine 6.0 8% Trehalose [F5] 20 mM histidine 6.0 / [surface] [27.] Stability study plan for excipient screening. F# stress condition T0 Sampling points and analysis [F1] [、] [F2] [、] [F3] [、] [F4] [and] [F5] [hot] 25℃ X 2W 4W X X 40℃ 1W 2W 4W X X X [Freezing and thawing] -40℃ to RT 3C 5C X X X = Appearance, pH, protein concentration, SEC-HPLC, cIEF, Caliper-SDS, DAR [3.4] [Sample Preparation]

[0207] FOR46 was buffer-exchanged to 20 mM histidine (pH 6.0) via ultrafiltration. The protein concentration was adjusted to 10 mg / mL after the addition of appropriate amounts of sucrose, trehalose, Arg-HCl, or NaCl. All samples were then aseptically filtered using a 0.22 µm PES membrane filter. For each formulation, eight (8) 2R glass vials were filled with 1 mL of filtered DS. One (1) vial underwent three and five freeze-thaw stress cycles, respectively. During each cycle, the vials were frozen at -40°C for at least 12 hours. The samples were thawed at room temperature. Three (3) vials were incubated at 40°C. Two vials were incubated at 25°C. One vial from each study condition was sampled at a specified time point for analysis. One (1) vial was used as T0. [3.5] [Results and Discussion] [] [3.5.1] [Appearance, protein concentration, and] [pH] [result]

[0208] Immediately after brief storage at 5°C, significant precipitation was observed in both F1 and F2, likely due to the high ionic strength of the formulation. Therefore, F1 and F2 were excluded from the study. At the outset of the study, all remaining portions of the sample were colorless, slightly milky white, and contained no visible ions.

[0209] After incubation at 25°C for up to 4 weeks for 40 seconds, no visible particles were observed in F5, while numerous particles were observed in F3 and F4. This may be attributed to protein denaturation induced by the higher surface tension of the sugar-containing formulation, and the aforementioned adverse effects can be eliminated by adding surfactants to the final formulation.

[0210] No substantial changes in appearance were found in F3, F4 and F5 after up to 5 cycles of freeze-thaw stress processes.

[0211] No substantial changes in pH or protein concentration were observed after five freeze-thaw cycles at 40°C and 25°C. [SEC] [purity]

[0212] SEC purity data are summarized in Table 28. Based on SEC data, no substantial changes were observed in any sample after a freeze-thaw cycle at 25°C for up to five cycles. After four weeks of incubation at 40°C, the SEC purity of F5 was significantly lower than that of F3 and F4. This demonstrates that the stabilizing effects of sucrose and trehalose on ADCs are unexpectedly significant and comparable. [surface] [28.] SEC purity results of FOR46 excipient screening study. Sample Information T0 FT 25℃ 40℃ 3C 5C 2W 4W 1W 2W 4W F3 Main Peak 97.9 98.6 98.5 97.5 98.1 96.0 93.2 94.9 HMW% 2.0 1.3 1.3 2.3 1.6 3.8 4.6 4.22 LMW% 0.1 0.2 0.2 0.1 0.3 0.2 2.1 0.92 F4 Main Peak 97.9 98.6 98.5 97.5 98.1 95.9 93.2 94.6 HMW% 2.0 1.2 1.3 2.3 1.6 3.9 4.8 4.4 LMW% 0.1 0.2 0.2 0.2 0.3 0.3 2.0 0.9 F5 Main peak% 97.5 98.4 98.4 96.8 97.8 94.7 9​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​[purity]

[0213] No substantial change in the purity of Caliper-SDS_R / NR was found in any sample after up to 5 freeze-thaw cycles and 4 weeks of incubation at 25°C and 40°C. [cIEF]

[0214] Based on cIEF data, after 4 weeks of cultivation at 40℃ and 25℃, a significant increase in the purity of the main peak was observed in all samples, with a similar rate of decrease in F3-F5. No substantial changes were observed after a maximum of 5 freeze-thaw cycles. [Drug-antibody ratio] [(DAR)]

[0215] After 4 weeks and up to 5 freeze-thaw cycles at 40°C and 25°C, no substantial changes in DAR were found in any of the samples. [in conclusion]

[0216] Even the worst appearance observed in buffers containing trehalose and sucrose was reversed by the addition of surfactants, despite the adverse effects caused by higher surface tension. Surprisingly, SEC purity results showed that sucrose and trehalose exhibited excellent and similar performance in FOR46 stabilization against thermal stress. Considering commercial costs, sucrose was selected as the excipient for optimizing the formulation. Surfactant screening studies will be conducted in a 20 mM histidine buffer (pH 6.0) (F3) containing 8% (w / v) sucrose. [4.] [Surfactant Screening]

[0217] This study aimed to evaluate the stabilizing effects of two different surfactants (PS-80 and PS-20) in 20 mM histidine buffer containing 8% (w / v) sucrose at three different concentrations. Based on DAR data (presented in Table 34), no substantial DAR changes were observed in any samples after 4 weeks and up to 5 freeze-thaw cycles at 40°C and 25°C. [Research Parameters]

[0218] FOR46 was prepared at a concentration of 10 mg / mL in 20 mM histidine buffer (pH 6.0) containing 8% (w / v) sucrose to a final concentration as follows: [surface] Of the seven formulations given in

[29] , PS-80 or PS-20 at three different concentration levels were added to each formulation, including a surfactant-free formulation as a blank control. Each formulation was subjected to up to five cycles of freeze-thaw cycles, thermal stress (40°C), and stirring stress (300 rpm, 2 days). The stability of the ADC at specified time points was assessed using different analyses. [surface] [29.] FOR46 Excipient Screening and Formulation Selection [Ingredient Number] [pH / ] [Buffer] [excipient] [Surfactant Notes] 1 20 mM His, pH 6.0 8% sucrose NA 2 0.01% PS-80 3 0.02% PS-80 4 0.03% PS-80 5 0.015% PS-20 6 0.02% PS-20 7 0.03% PS-20 [surface]

[30] [.] FOR46 Excipient Screening and Formulation Selection [characteristic] [condition] [T0] [Sampling Points and Analysis] [hot] 40℃ X, Y, Z 2W 4W X X, Z [freeze] [ / ] [melt] -40℃ to RT 5 cycles X, Y, Z [Stir] 25℃, 300 rpm 2D X, Y, Z X = Appearance, pH, protein concentration (SEC-HPLC, cIEF, SDS caliper_R); Y = MFI; Z = Combined stirring [4.3] [Drug Materials]

[0219] Before the surfactant screening study, FOR46, formulated in 20 mM histidine buffer (pH 6.0) with 8% (w / v) sucrose, was stored at 2-8°C. [Sample Preparation]

[0220] After adding the specified amount of PS-80 or PS-20, the WBP2O95 ADC DS was aseptically filtered using a 0.22-µm PES membrane filter. For each formulation sample, eight (8) 2R glass vials were each filled with 1 mL of filtered DS. Two (2) vials were subjected to five cycles of freeze-thaw stress. In each cycle, the freezing time was at least 12 hours in a -40°C freezer. The samples were thawed at room temperature. The two (2) vials were incubated at 40°C. At ambient temperature, the two (2) vials were subjected to stirring at 300 rpm for 2 days. One vial from 40°C and two vials from the freeze-thaw stress and stirring stress processes were sampled at specified time points for analysis. The two (2) vials were used as T0. [4.5] [Results and Discussion] [] [4.5.1] [Appearance, protein concentration, and] [pH] [result]

[0221] No substantial changes in appearance were observed in any of the samples after five freeze-thaw cycles. Particles and fibers were observed in F1 (surfactant-free) after stirring at 300 rpm for two days and incubating at 40°C for four weeks. This indicates that the presence of surfactants may be crucial for protecting the ADC under thermal and stirring stress conditions.

[0222] No substantial changes in pH or protein concentration were found. [4.5.2 SEC] [purity]

[0223] No substantial change in SEC purity was observed after 5 cycles of freeze-thaw cycles and 2 days of stirring. After 4 weeks of incubation at 40°C, a 6% decrease in the purity of the main peak was observed in all 7 formulations. Based on SEC purity data, all formulations were similar under all conditions. [4.5.3 CE-SDS_R] [purity]

[0224] No substantial changes in CE-SDS R purity were found under thermal stress, freeze-thaw stress, and stirring stress conditions. [4.5.4 cIEF]

[0225] No substantial changes in cIEF were observed after five cycles of freeze-thaw cycles and two days of stirring. Under thermal stress, the purity of the main peak decreased significantly, while the purity of the acid peak increased accordingly. However, the changes were similar in all formulations. [4.5.5] [efficacy]

[0226] Based on previous data, three main formulations (F2, F3, and F4) were selected for binding performance analysis. No substantial changes in binding performance were found under thermal stress, stirring stress, and freeze-thaw stress. [4.5.6 MFI]

[0227] Surprisingly, based on the MFI results, more than 10 times more particles were found in F1 compared to the other formulations. This indicates that F1 has more sub-visible particles than other formulations. [surface] [31.] MFI results of FOR46 in the surfactant screening study. sample T0 FT Stir F1 2~5 um 7085 2561 4874 5~10 um 1340 523 1337 10~25 um 194 79 478 ≥25 um 15 4 69 F2 2~5 um 542 368 404 5~10 um 104 51 33 10~25 um twenty two 9 5 ≥25 um 0 0 0 F3 2~5 um 1467 340 258 5~10 μm 379 35 15 10-25 um 94 10 4 ≥25 um 2 0 0 F4 2-5 units 545 969 332 5-10 one 53 109 56 10-25 um 7 12 10 ≥25 um 4 0 2 F5 2-5 units 692 716 1050 5-10 one 84 60 99 10-25 um 10 9 4 ≥25 um 0 0 0 F6 2-5 units 550 337 294 5-10 one 63 30 25 10-25 um 20 12 10 ≥25 um 5 2 4 F7 2~5 um 813 965 689 5~10 μm 114 171 130 10~25 um 30 33 78 ≥25 um 30 33 78 [4.6] [in conclusion]

[0228] Based on appearance and MFI results, the surfactant played an unexpectedly important role in protecting the ADC under thermal and stirring stress conditions. However, no differences were found among the six formulations containing two different surfactants (PS-80 and PS-20) at three concentration levels. Considering that PS-80 has a lower CMC (critical microcell concentration) than PS-20, indicating a lower effective concentration of the surfactant, and taking into account the potential adverse effects introduced by the decomposition of PS-80 at high concentration levels, 0.01% (w / v) PS-80 was selected in the final formulation.

[0229] FOR46 (10 mg / mL) was selected in 20 mM histidine buffer (pH 6.0) with 8% (w / v) sucrose and 0.01% (w / v) PS-80 as the final formulation.

[0230] <![CDATA[ <110> FORTIS THERAPEUTICS, INC. (USA) <![CDATA[ <120> CD46-targeting immune conjugates and their usage]]> <![CDATA[ <140> TW 110129203]]> <![CDATA[ <141> 2021-08-06 <![CDATA[ <150> US 63 / 062,740 <![CDATA[ <151> 2020-08-07 <![CDATA[ <160> 10 ]]> <![CDATA[ <170> PatentIn version 3.5]]> <![CDATA[ <210> 1]]> <![CDATA[ <211> 8]]> <![CDATA[ <212> PRT]]> <![CDATA[ <213> Artificial sequence <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Description of Artificial Sequences: Synthetic Peptides"]]> <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Synthetic antibody VH CDR1 domain"]]> <![CDATA[ <400> 1]]> Gly Leu Thr Val Asn Asn Tyr Ala 1 5 <![CDATA[ <210> 2]]> <![CDATA[ <211> 8]]> <![CDATA[ <212> PRT]]> <![CDATA[ <213> Artificial sequence <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Description of Artificial Sequences: Synthetic Peptides"]]> <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Synthetic antibody VH CDR2 domain"​​]]> <![CDATA[ <400> 2]]> Ile Ser Tyr Asp Gly Asn Asn Lys 1 5 <![CDATA[ <210> 3]]> <![CDATA[ <211> 9]]> <![CDATA[ <212> PRT]]> <![CDATA[ <213> Artificial sequence <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Description of Artificial Sequences: Synthetic Peptides"]]> <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Synthetic antibody VH CDR3 domain"]]> <![CDATA[ <400> 3]]> Ala Lys Gly Gly Gly Tyr Phe Asp Leu 1 5 <![CDATA[ <210> 4]]> <![CDATA[ <211> 9]]> <![CDATA[ <212> PRT]]> <![CDATA[ <213> Artificial sequence <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Description of Artificial Sequences: Synthetic Peptides"]]> <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Synthetic antibody VL CDR1 domain"]]> <![CDATA[ <400> 4]]> Ser Ser Asn Ile Gly Ala Gly Tyr Asp 1 5 <![CDATA[ <210> 5]]> <![CDATA[ <211> 3]]> <![CDATA[ <212> PRT]]> <![CDATA[ <213> Artificial sequence <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Description of Artificial Sequences: Synthetic Peptides"]]> <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Synthetic antibody VL CDR2 domain"​​]]> <![CDATA[ <400> 5]]> Gly Asn Asn 1 <![CDATA[ <210> 6]]> <![CDATA[ <211> 9]]> <![CDATA[ <212> PRT]]> <![CDATA[ <213> Artificial sequence <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Description of Artificial Sequences: Synthetic Peptides"]]> <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Synthetic antibody VL CDR3 domain"]]> <![CDATA[ <400> 6]]> Ser Ser Tyr Thr Ser Gly Thr Trp Leu 1 5 <![CDATA[ <210> 7]]> <![CDATA[ <211> 116]]> <![CDATA[ <212> PRT]]> <![CDATA[ <213> Artificial sequence <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Description of Artificial Sequences: Synthetic Polypeptides"]]> <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Synthetic antibody VH domain"]]> <![CDATA[ <400> 7]]> Gln Val Gln Leu Val Gln Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ala Cys Ala Ala Ser Gly Leu Thr Val Asn Asn Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Ser Tyr Asp Gly Asn Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Gly Gly Tyr Phe Asp Leu Trp Gly Arg Gly Thr Leu Val 100 105 110 Thr Val Ser Ser 115 <![CDATA[ <210> 8]]> <![CDATA[ <211> 109]]> <![CDATA[ <212> PRT]]> <![CDATA[ <213> Artificial sequence <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Description of Artificial Sequences: Synthetic Polypeptides"]]> <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Synthetic antibody VL domain"]]> <![CDATA[ <400> 8]]> Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Gly 20 25 30 Tyr Asp Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu 35 40 45 Leu Ile Tyr Gly Asn Asn Asn Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Gly 85 90 95 Thr Trp Leu Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 <![CDATA[ <210> 9]]> <![CDATA[ <211> 446]]> <![CDATA[ <212> PRT]]> <![CDATA[ <213> Artificial sequence <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Description of Artificial Sequences: Synthetic Polypeptides"]]> <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Synthetic antibody H domain"]]> <![CDATA[ <400> 9]]> Gln Val Gln Leu Val Gln Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ala Cys Ala Ala Ser Gly Leu Thr Val Asn Asn Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Ser Tyr Asp Gly Asn Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Gly Gly Tyr Phe Asp Leu Trp Gly Arg Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 165 170 175 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 210 215 220 Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe 225 230 235 240 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 245 250 255 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 260 265 270 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 290 295 300 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 340 345 350 Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 355 360 365 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 370 375 380 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 385 390 395 400 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 405 410 415 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 420 425 430 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <![CDATA[ <210> 10]]> <![CDATA[ <211> 215]]> <![CDATA[ <212> PRT]]> <![CDATA[ <213> Artificial sequence <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Description of Artificial Sequences: Synthetic Polypeptides"]]> <![CDATA[ <220> ]]> <![CDATA[ <221> Source <![CDATA[ <223> / note="Synthetic antibody L-domain"]]> <![CDATA[ <400> 10]]> Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln 1 5 10 15 Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Gly 20 25 30 Tyr Asp Val His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu 35 40 45 Leu Ile Tyr Gly Asn Asn Asn Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Gly 85 90 95 Thr Trp Leu Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln Pro 100 105 110 Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu 115 120 125 Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr Pro 130 135 140 Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys Ala 145 150 155 160 Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr Ala 165 170 175 Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg 180 185 190 Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys Thr 195 200 205 Val Ala Pro Thr Glu Cys Ser 210 215

[0231]

Claims

1. An immunoconjugate comprising: a recombinant antibody comprising: a first heavy chain comprising SEQ ID NO: 9, a first light chain comprising SEQ ID NO: 10, a second heavy chain comprising SEQ ID NO: 9, and a second light chain comprising SEQ ID NO: 10; and one, two, three, or four adducts; wherein each of the one, two, three, or four adducts comprises monomethylauristatin E (MMAE), which is conjugated to the recombinant antibody via a maleiminohexyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB) linker; wherein each of the one, two, three, or four adducts is conjugated to one pair of cysteine ​​residues of the recombinant antibody, wherein the cysteine ​​residue pairs are selected from: C219 of the first heavy chain and C214 of the first light chain; The second heavy chain C219 and the second light chain C214; the first heavy chain C225 and the second heavy chain C225; and the first heavy chain C228 and the second heavy chain C228.

2. An immune conjugate comprising: a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region including three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3; and a light chain (LC) variable region including three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 respectively contain the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, and LC CDR1, LC CDR2, and LC CDR3 respectively contain the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6; and one, two, three, or four adducts; wherein each of the one, two, three, or four adducts comprises monomethylaurestatin E. (MMAE), which is conjugated to the recombinant antibody via a maleiminohexylacetyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB) linker; wherein each of the first, second, third, or fourth adduct pairs is conjugated to one pair of cysteine ​​residues of the recombinant antibody, wherein the cysteine ​​residue pairs are selected from: C219 of the first heavy chain and C214 of the first light chain; C219 of the second heavy chain and C214 of the second light chain; C225 of the first heavy chain and C225 of the second heavy chain; and C228 of the first heavy chain and C228 of the second heavy chain.

3. A pharmaceutical composition comprising an immunoconjugate, a pharmaceutically acceptable buffer, and a pharmaceutically acceptable stabilizer; wherein the immunoconjugate comprises a recombinant antibody comprising a first heavy chain comprising SEQ ID NO: 9, a first light chain comprising SEQ ID NO: 10, a second heavy chain comprising SEQ ID NO: 9, and a second light chain comprising SEQ ID NO: 10; and one, two, three, or four adducts; wherein each of the one, two, three, or four adducts comprises monomethylauristatin E (MMAE), which is conjugated to the recombinant antibody via a maleiminohexyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB) linker; wherein each of the one, two, three, or four adducts is conjugated to one pair of cysteine ​​residues of the recombinant antibody, wherein the cysteine ​​residue pairs are selected from: The first heavy chain C219 and the first light chain C214; the second heavy chain C219 and the second light chain C214; the first heavy chain C225 and the second heavy chain C225; and the first heavy chain C228 and the second heavy chain C228.

4. A pharmaceutical composition comprising an immune conjugate, a pharmaceutically acceptable buffer, and a pharmaceutically acceptable stabilizer; wherein the immune conjugate comprises: a recombinant antibody that specifically binds to CD46, comprising a heavy chain (HC) variable region comprising three complementarity-determining regions (CDRs): HC CDR1, HC CDR2, and HC CDR3; and a light chain (LC) variable region comprising three CDRs: LC CDR1, LC CDR2, and LC CDR3, wherein HC CDR1, HC CDR2, and HC CDR3 comprise the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and LC CDR1, LC CDR2, and LC CDR3 comprise the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively; and one, two, three, or four adduct pairs; Each of the first, second, third, or fourth adduct pairs comprises monomethylaurestatin E (MMAE), which is conjugated to the recombinant antibody via a maleiminohexyl-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB) linker; wherein each of the first, second, third, or fourth adduct pairs is conjugated to one pair of cysteine ​​residues of the recombinant antibody, wherein the cysteine ​​residue pairs are selected from: C219 of the first heavy chain and C214 of the first light chain; C219 of the second heavy chain and C214 of the second light chain; C225 of the first heavy chain and C225 of the second heavy chain; and C228 of the first heavy chain and C228 of the second heavy chain.

5. The pharmaceutical composition of claim 3 or 4, wherein the pharmaceutical composition has a pH of about 5.0 to about 7.

0.

6. The pharmaceutical composition of claim 3 or 4, wherein the buffer comprises citrate, phosphate, acetate, tromethamine, histidine, succinate, malate, or α-ketoglutarate.

7. The pharmaceutical composition of claim 3 or 4, wherein the buffer contains about 10 mM to about 30 mM histidine and has a pH of about 5 to about 7.

8. The pharmaceutical composition of claim 6, wherein the buffer contains about 20 mM histidine and has a pH of about 6.

0.

9. The pharmaceutical composition of claim 3 or 4, wherein the stabilizer comprises polysorbate.

10. The pharmaceutical composition of claim 3 or 4 further comprises a medically acceptable cryoprotectant.

11. The pharmaceutical composition of claim 10, wherein the cryoprotectant comprises sugar.

12. The pharmaceutical composition of claim 4, wherein the ratio of the effector to the recombinant antibody in the group of immune conjugates is about 3 to about 5.

13. The pharmaceutical composition of claim 4, wherein the effector is conjugated to the recombinant antibody via a linker.

14. The pharmaceutical composition of claim 13, wherein the linker comprises maleiminohexylidene-valine-citrulline-p-aminobenzoxycarbonyl (mc-vc-PAB).

15. Use of an immune conjugate as claimed in claim 1 or 2 or a pharmaceutical composition as claimed in any one of claims 3 to 14 for the preparation of a pharmaceutical product, wherein the pharmaceutical product is for the treatment of cancer in a human individual containing cells expressing CD46.

16. As claimed in claim 15, wherein the cancer is selected from relapsed or refractory multiple myeloma (RRMM) and metastatic castration-resistant prostate cancer (mCRPC).

17. As claimed in claim 15 or 16, wherein the treatment further comprises detecting the CD46 in the cell.

18. As claimed in claim 17, wherein the detection includes immunofluorescence microscopy, immunohistochemistry, flow cytometry, or a combination thereof.

19. As requested in claim 18, wherein the detection includes the detection of amplification of chromosome position 1q21.

20. The use as claimed in claim 15 or 16, wherein the medicine is administered to the human individual via intravenous infusion.

21. As claimed in paragraphs 15 or 16, wherein the medicine is administered to the human individual every 7 days, every 14 days, every 18 days, every 21 days, or every 30 days.

22. The use as claimed in claim 15 or 16, wherein the pharmaceutical product is administered to the immune conjugate or pharmaceutical composition at a dose of about 1.2 to about 3.0 mg / kg.

23. As used in claim 22, wherein the kg weight of the human individual is the actual body weight.

24. As used in claim 22, wherein the kg weight of the human individual is the adjusted weight (AJBW).

25. As claimed in claim 24, wherein the AJBW is: For male human individuals, AJBW = IBW + 0.4 × (actual weight – IBW); wherein IBW is ideal weight, which is: For male human individuals, IBW = 50 kg + 2.3 kg × (actual height – 60 inches); and for female human individuals, IBW = 45.5 kg + 2.3 kg × (actual height – 60 inches); wherein the AJBW, the IBW, and the actual weight are measured in kg, and the actual height is measured in inches.

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