WO2025252894A1 - Molécule de liaison à ca19-9 - Google Patents

Molécule de liaison à ca19-9

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Publication number
WO2025252894A1
WO2025252894A1 PCT/EP2025/065686 EP2025065686W WO2025252894A1 WO 2025252894 A1 WO2025252894 A1 WO 2025252894A1 EP 2025065686 W EP2025065686 W EP 2025065686W WO 2025252894 A1 WO2025252894 A1 WO 2025252894A1
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WIPO (PCT)
Prior art keywords
amino acid
seq
acid sequence
binding molecule
bpc
Prior art date
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Pending
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PCT/EP2025/065686
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English (en)
Inventor
Ugur Sahin
Ursula ELLINGHAUS
Kathrin RÖSCH
Magdalena UTSCH
Christiane STADLER
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Biontech SE
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Biontech SE
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Publication of WO2025252894A1 publication Critical patent/WO2025252894A1/fr
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A61K47/68037—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a camptothecin [CPT] or derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6851—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell
    • A61K47/6859—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell the tumour determinant being from liver or pancreas cancer cell
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • C07K16/3046—Stomach, Intestines
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/44—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material not provided for elsewhere, e.g. haptens, metals, DNA, RNA, amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/71—Decreased effector function due to an Fc-modification
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/77—Internalization into the cell

Definitions

  • BPCs binding molecule-payload conjugates
  • ADCs antibody-drug conjugates
  • Sialyl Lewis A (sLe a ) antigen is an epitope present on Carbohydrate Antigen 19-9 (CA19- 9), that has been shown to be overexpressed on epithelial cell tumors (Magnani et al., 1982, J Biol Chem.257:14365-14369; Magnani et al., 1983, Cancer Res.43:5489-5492).
  • sLe a is an 10 oligosaccharide expressed primarily as a proteoglycan that is secreted and circulates as a mucin form, and also as a less well studied glycolipid form (Magnani et al., 1983, Cancer Res.
  • sLe a antigen is expressed predominantly on cancer cells (Kannagi et al., 2007, Chang Gung Med J.30:189-209).
  • sLe a facilitates tumor adhesion and extravasation, key events for tumor 15 metastasis, and is thus a marker of an aggressive tumor phenotype (Sato et al., 1997, Anticancer Res.17:3505-3511).
  • Glycolipids, such as sLe a are established targets for cancer immunotherapies (Feizi, 1985, Nature 314:53-57).
  • CA19-9 is widely expressed on tumors of the gastrointestinal tract, with up to 94% of pancreatic cancers positive for CA19-9 expression and high expression rates also seen in bile duct carcinomas and transitional cell carcinomas 20 (Loy et al., 1993, Am J Clin Pathol. 99:726-728; Passerini et al., 2012, Am J Clin Pathol 138:281-287). Additionally, expression of CA19-9 is frequently seen in ovarian, colon, stomach, and distal esophagus/stomach cancers.
  • Circulating serum levels of CA19-9 have been validated as a biomarker for assessing the metastatic potential of pancreatic ductal adenocarcinomas (PDAC) (Ballehaninna and 25 Chamberlain, 2012, J Gastrointest Oncol. 3(2):105-119; Dong, 2014, World J Surg Oncol. 12:171) and have been used to evaluate the aggressiveness of other epithelial cell cancers (Locker et al., 2006, J Clin Oncol.24:5313-5327; Nakayama, 1995, Cancer 75:2051-2056).
  • PDAC pancreatic ductal adenocarcinomas
  • CA19-9 expression is associated with increased metastatic potential in colon cancer (Matsui et al., 2004, Jpn J Clin 30 Oncol.34:588-593; Ben-David, 2008, Immunol Lett.116:218-224; Sato et al., 1997, Anticancer Res. 17:3505-3511) and pancreatic adenocarcinoma (Kishimoto et al., 1996, Int J Cancer 69:290-294).
  • Antibody-drug conjugates comprise biologically active small molecule compounds 15 conjugated to monoclonal antibodies or antibody fragments by chemical methods, so as to fully utilize antibodies’ binding specificity to normal cell and to tumor cell surface antigens, and small molecule’s high anti-tumor biological activity, while avoiding defects such as the low specific efficacy of the former as well as toxic side effects of the latter.
  • antibody-drug conjugates can more accurately 20 bind to tumor cells and reduce their effects on normal cells.
  • the present invention provides a binding molecule-payload conjugate (BPC) comprising a 25 binding molecule and one or more payload moieties; wherein: a) the binding molecule is an antibody or a fragment thereof and comprises heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3 according to the IMGT numbering scheme, wherein the binding molecule specifically binds to CA19-9; and 30 b) the one or more payload moieties is covalently linked to the binding molecule.
  • the one or more payload moieties is covalently linked to the binding P129624PCT molecule via one or more linkers.
  • the BPC according to the invention comprises at least one of HCDRs 1-3 and/or LCDRs 1-3 selected from: a) HCDR1 comprising an amino acid sequence according to SEQ ID NO: 1, 5 b) HCDR2 comprising an amino acid sequence according to SEQ ID NO: 2, c) HCDR3 comprising an amino acid sequence according to SEQ ID NO: 3, d) LCDR1 comprising an amino acid sequence according to SEQ ID NO: 4, e) LCDR2 comprising an amino acid sequence according to SEQ ID NO: 5, and f) LCDR3 comprisng an amino acid sequence according to SEQ ID NO: 6.
  • the invention provides a binding molecule-payload conjugate (BPC) comprising a binding molecule and one or more payload moieties; wherein: a) the binding molecule is an antibody or a fragment thereof and comprises heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity 30 determining regions (LCDRs) 1-3 according to the Kabat numbering scheme, wherein: P129624PCT HCDR1 comprises an amino acid sequence according to SEQ ID NO: 15, HCDR2 comprises an amino acid sequence according to SEQ ID NO: 16, HCDR3 comprises an amino acid sequence according to SEQ ID NO: 17, LCDR1 comprises an amino acid sequence according to SEQ ID NO: 18, LCDR2 comprises an amino acid 5 sequence according to SEQ ID NO: 19, LCDR3 comprises an amino acid sequence according to SEQ ID NO: 6, optionally wherein one or more of the CDRs comprises one, two or three amino acid mutations relative to the recited sequences; and b) the one or more payload moieties is covalently linked to
  • the invention provides a binding molecule-payload conjugate (BPC) comprising a binding molecule and one or more payload moieties; wherein: a) the binding molecule is an antibody or a fragment thereof and comprises heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3 according to the Chothia numbering scheme, 15 wherein: HCDR1 comprises an amino acid sequence according to SEQ ID NO: 20, HCDR2 comprises an amino acid sequence according to SEQ ID NO: 21, HCDR3 comprises an amino acid sequence according to SEQ ID NO: 17, LCDR1 comprises an amino acid sequence according to SEQ ID NO: 18, LCDR2 comprises an amino acid 20 sequence according to SEQ ID NO: 19, LCDR3 comprises an amino acid sequence according to SEQ ID NO: 6, optionally wherein one or more of the CDRs comprises one, two or three amino acid mutations relative to the recited sequences; and b) the one or more payload moieties is covalently linked to the binding molecule,
  • the binding molecule according to the invention may comprise a heavy chain variable region (VH) comprising an amino acid sequence according to SEQ ID NO: 7 or a variant having at least 80% sequence identity thereto, and/or a light chain variable region (VL) comprising an amino acid sequence according to SEQ ID NO: 8 or a variant having at least 80% sequence identity thereto.
  • VH heavy chain variable region
  • VL light chain variable region
  • the binding molecule further comprises an Fc region.
  • the Fc region may be a modified Fc region.
  • the payload moiety is selected from a drug, a detectable marker, a 5 radioisotope, a fluorescent agent, a luminescent agent, a colored agent, an enzyme, polyethylene glycol, a nuclide, a nucleic acid, a small molecule toxin, a polypeptide having binding activity, a protein, a receptor, a ligand, another active agent that inhibits tumor cell growth, promotes tumor cell apoptosis or necrosis.
  • the payload moiety is a drug.
  • the drug may be a cytotoxic drug, 10 immune modulator, or a STING inhibitor.
  • the payload-linker moiety of the conjugate may have the following structure: .
  • the payload-linker comprises a structure shown as formula (II-A): P129624PCT or a tautomer, a mesomer, a racemate, an enantiomer or a diastereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, X 1 is saturated C, and X 1 is substituted with R n ;
  • 5 ring A is selected from the group consisting of: 3-10 membered saturated or partially unsaturated heterocyclyl and 3-10 membered saturated or partially unsaturated carbocyclyl, wherein ring A is substituted with 0 or at least 1 substituent R 1a ; when ring A is 3-10 membered saturated or partially unsaturated carbocyclyl, ring A is substituted with p L 2 , and L 2 is not R n ; 10 or, when ring A is 3-10 membered saturated or partially unsaturated heterocycl
  • the payload-linker moiety may have the structure: .
  • the conjugate has the structure represented by formula I: 15 P129624PCT or a tautomer, a mesomer, a racemate, an enantiomer or a diastereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, BM is the binding molecule; 5 each Z is independently selected from a direct bond, a carbon-carbon triple bond, a carbon- carbon double bond, and amido (preferably selected from a direct bond, a carbon- 10 carbon triple bond, and a carbon-carbon double bond); Rx and Ry are each independently selected from H and C1-4 alkyl; each m is independently selected from 0, 1, 2, 3, 4, 5 and 6; y1 is selected from any integer between 1 and 6 (such as 4, 5, and 6); each y2 is independently selected from any integer between 0 and 15 (such as 6-15); 15 each y3 is independently selected from 1, 2, and 3; each y4 is independently selected from 0 and 1; position
  • R r, r 1 , r 1a and r 1b are each independently 0, 1, 2, 3, 4 or 5;
  • R m1 , R n1 , R m1a , R n1a , R m1b and R n1b are each independently H, C 1-6 alkyl or -COOR x1 , wherein, R x1 is C 1-6 alkyl; or, R m1 and R n1 , R m1a and R n1a , and R m1b and R n1b , together with the nitrogen atom to which 20 they are both attached, form a 5-6 membered heterocyclic ring, and of said 5-6 membered heterocyclic ring, the heteroatom is selected from 1 or 2 N atoms; the said 5-6 membered heterocyclic ring is optionally substituted with one or more R 0’ ;
  • R z is selected from C 1-6 alkyl;
  • the linker-payload moiety may comprise the structure: 5
  • the invention also provides an antibody comprising: a) heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3 according to the IMGT numbering scheme, wherein: HCDR1 comprises an amino acid sequence according to SEQ ID NO: 1, HCDR2 10 comprises an amino acid sequence according to SEQ ID NO: 2, HCDR3 comprises an amino acid sequence according to SEQ ID NO: 3, LCDR1 comprises an amino acid sequence according to SEQ ID NO: 4, LCDR2 comprises an amino acid sequence according to SEQ ID NO: 5, LCDR3 comprises an amino acid sequence according to SEQ ID NO: 6, optionally wherein one or more of the CDRs comprises one, two or 15 three amino acid mutations relative to the recited sequences; and b) a modified Fc region.
  • HCDR1 comprises an amino acid sequence according to SEQ ID NO: 1
  • HCDR2 10 comprises an amino acid sequence according to SEQ ID NO: 2
  • the invention provides an antibody comprising: a) heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3 according to the Kabat numbering 20 scheme, wherein: HCDR1 comprises an amino acid sequence according to SEQ ID NO: 15, HCDR2 comprises an amino acid sequence according to SEQ ID NO: 16, HCDR3 comprises an amino acid sequence according to SEQ ID NO: 17, LCDR1 comprises an amino acid sequence according to SEQ ID NO: 18, LCDR2 comprises an amino acid P129624PCT sequence according to SEQ ID NO: 19, LCDR3 comprises an amino acid sequence according to SEQ ID NO: 6, optionally wherein one or more of the CDRs comprises one, two or three amino acid mutations relative to the recited sequences; and b) a modified Fc region.
  • HCDR1 comprises an amino acid sequence according to SEQ ID NO: 15
  • HCDR2 comprises an amino acid sequence according to SEQ ID NO: 16
  • HCDR3 comprises an amino acid sequence according to S
  • the invention provides an antibody comprising: a) heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3 according to the Chothia numbering scheme, wherein: HCDR1 comprises an amino acid sequence according to SEQ ID NO: 20, HCDR2 10 comprises an amino acid sequence according to SEQ ID NO: 21, HCDR3 comprises an amino acid sequence according to SEQ ID NO: 17, LCDR1 comprises an amino acid sequence according to SEQ ID NO: 18, LCDR2 comprises an amino acid sequence according to SEQ ID NO: 19, LCDR3 comprises an amino acid sequence according to SEQ ID NO: 6, optionally wherein one or more of the CDRs comprises 15 one, two or three amino acid mutations relative to the recited sequences; and b) a modified Fc region.
  • HCDR1 comprises an amino acid sequence according to SEQ ID NO: 20
  • HCDR2 10 comprises an amino acid sequence according to SEQ ID NO: 21
  • HCDR3 comprises an amino acid sequence according to SEQ ID NO
  • the antibody may comprise a heavy chain variable region (VH) comprising an amino acid sequence according to SEQ ID NO: 7 or a variant having at least 80% sequence identity thereto.
  • the antibody may comprise a light chain variable region (VL) comprising an amino 20 acid sequence according to SEQ ID NO: 8 or a variant having at least 80% sequence identity thereto.
  • the Fc region comprises an amino acid substitution at position 234, or an amino acid substitution at position 235, or an amino acid substitution at position 236, or an amino acid substitution at positions 234 and 235, or an amino acid substitution at positions 25 234, 235 and 236, according to the EU numbering scheme.
  • the one or more amino acid substitution(s) may be selected from the group consisting of: a) L234A, L235A, A327G, A330S, and P331S (LALA- ⁇ A); b) L234A, L235A, and K322A (LALA-KA); c) L234S, L235T, and G236R (STR); and 30 d) L234A and L235A (LALA).
  • the invention also provides a method for producing a BPC according to the invention, P129624PCT comprising contacting an antibody according to the invention with a suitable linker-payload compound.
  • the invention also provides one or more nucleic acid sequence(s) encoding a binding molecule as defined herein or an antibody according the invention.
  • the one or more nucleic 5 acid sequence(s) may be an RNA sequence.
  • the invention provides a BPC according to the invention for use in a method of therapy or a diagnostic method.
  • the invention provides an antibody according to the invention for use in method of therapy or a diagnostic method.
  • the method may be a method of treating, preventing or diagnosing cancer.
  • the cancer may express CA19-9.
  • the cancer is gastrointestinal cancer, pancreatic cancer, ovarian cancer, colorectal cancer, stomach cancer, oesophageal cancer, endometrial cancer, breast cancer, bile duct carcinoma, transitional cell carcinoma, or hepatocellular carcinoma.
  • the cancer is pancreatic ductal adenocarcinoma (PDAC).
  • Hsd:Athymic Nude-Foxn1 nu/nu mice (n 14) carrying a s.c. human pancreatic ductal adenocarcinoma (BxPC3) xenograft were treated q1w with in total of seven i.p. doses of 3 or 8 mg/kg 5B1-ADC, 8 mg/kg ctrl IgG-ADC or the naked 5B1 mAb. Tumor growth was monitored 25 three times a week.
  • Hsd:Athymic Nude-Foxn1 nu/nu mice (n 13-14) carrying a s.c.
  • FIG. 10 Figure 9 – Cell cycle analysis of COLO 205 (A) and BxPC3 (B) cells 24 hours post-treatment with DXd, Fc-inert 5B1-ADC and ADC control.
  • the cell cycle of COLO 205 (A) and BxPC3 (B) cells was analyzed 24 h post treatment with 5B1-ADC Fc-inert or control items by measuring BrdU incorporation and DNA content via flow cytometry.
  • DXd served as positive and BNT- ADC-IC as negative control.
  • FIG 10 Cell apoptosis in COLO 205 (A) and BxPC3 (B) cells 48 hours post treatment with 5B1-ADC Fc-inert, ADC-IC and DXd.
  • Cell apoptosis was measured 48 h post treatment of COLO 205 (A) and BxPC3 (B) cells with 5B1-ADC Fc-inert or control items.
  • Figure 11 Bystander killing activity of 5B1-ADC Fc-inert.
  • Target-negative SK-OV-3 cells were co-cultured with target-positive COLO 205 (A) or BxPC3 cells (B) at a ratio of 1:3 for 5 days.
  • Figure 12 Tolerability and therapeutic efficacy of 5B1-ADC Fc-inert in a human pancreas carcinoma CDX model. The study animals were injected with test and control items on Day 34 and received a total of six dose administrations on a weekly basis (dotted lines in A and B).
  • 5B1- ADC Fc-inert at 3 and 8 mg/kg were compared to 5B1-mAb Fc-comp, 5B1-ADC Fc-comp and 5 DPBS. *p ⁇ 0.05, **p ⁇ 0.01, ***p ⁇ 0.001 ****p ⁇ 0.0001.
  • Figure 13 Tolerability and therapeutic efficacy of 5B1-ADC Fc-inert in a human colon carcinoma CDX model.
  • mice per group differed over the study period due to premature death or euthanasia in accordance with GV-SOLAS guidelines.
  • Figure 14 Tolerability and therapeutic efficacy of 5B1-ADC Fc-inert in a human pancreas ductal adenocarcinoma CDX model.
  • the study animals were injected with test and control items on Day 5 and received a total of four doses on a weekly basis (dotted lines).
  • On the day 20 of tumor cell inoculation, each group included 10 mice (n 10). However, the number of mice per group differed over the study period due to premature death or euthanasia in accordance with GV-SOLAS guidelines.
  • binding molecule-payload conjugate comprising a 30 binding molecule and one or more payload moieties, wherein the binding molecule specifically binds to CA19-9.
  • BPC binding molecule-payload conjugate
  • CA19-9 is considered the antigen of the binding molecules according to the present invention.
  • “specifically binds to” may indicate that the binding molecule binds to the antigen, i.e. CA19-9 in preference to other antigens.
  • “specifically binds to” refers to the antibody- like binding of the binding molecule, which may be via heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3, to the target CA19-9.
  • HCDRs heavy chain complementarity determining regions
  • LCDRs light chain complementarity determining regions
  • CA19-9 relates to Carbohydrate Antigen 19-9 and includes any modifications which can comprise the Sialyl Lewis A (sLe a ) antigen epitope.
  • the sLe a antigen may have the following structure:
  • the binding molecule does not show cross reactivity to sLe x , Le a and/or 15 Le Y antigens.
  • Suitable assays and techniques for measuring/quantifying binding activity of the binding molecule according to the invention may include, but are not limited to, ELISA, surface plasmon resonance (SPR), bio-layer interferometry (BLI), quartz crystal microbalance (QCM), bioluminescence assays and flow cytometry.
  • SPR surface plasmon resonance
  • BLI bio-layer interferometry
  • QCM quartz crystal microbalance
  • bioluminescence assays and flow cytometry.
  • Other suitable techniques will be known in the 20 art.
  • EC50 is a measure of the concentration of a binding molecule that induces a specific response that is 50% between the maximum response and the baseline response.
  • EC50 can be used to assess the ability of a binding molecule to bind to a target.
  • the binding molecule according to the present invention is an antibody or a fragment thereof.
  • P129624PCT The term “antibody” refers to a glycoprotein comprising at least two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds, and includes any molecule comprising an antigen binding portion thereof.
  • antibody includes monoclonal antibodies and fragments or derivatives of antibodies, including, without limitation, human antibodies, 5 humanized antibodies, chimeric antibodies, single chain antibodies, e.g., scFvs and antigen- binding antibody fragments such as Fab and Fab' fragments and also includes all recombinant forms of antibodies, e.g., antibodies expressed in prokaryotes, unglycosylated antibodies, and any antigen-binding antibody fragments and derivatives as described herein.
  • each heavy chain is comprised of a heavy chain variable region 10 (abbreviated herein as VH) and a heavy chain constant region
  • each light chain is comprised of a light chain variable region (abbreviated herein as VL) and a light chain constant region.
  • the VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more 15 conserved, termed framework regions (FR).
  • CDR complementarity determining regions
  • FR framework regions
  • Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.
  • variable regions of the heavy and light chains contain a binding domain that interacts with an antigen.
  • the constant regions of the antibodies may mediate the binding of the immunoglobulin to host tissues or factors, including various 20 cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system.
  • a CDR refers to one of three hypervariable regions (H1, H2 or H3) within the non-framework region of the immunoglobulin (Ig or antibody) VH ⁇ -sheet framework, or one of three hypervariable regions (L1, L2 or L3) within the non-framework region of the antibody VL ⁇ - 25 sheet framework. Accordingly, CDRs are variable region sequences interspersed within the framework region sequences.
  • CDR regions are well known to those skilled in the art and have been defined by, for example, Kabat as the regions of most hypervariability within the antibody variable (V) domains (Kabat et al., 1977, J. Biol. Chem. 252:6609-6616; Kabat, 1978, Adv. Prot. Chem.32:1-75). CDR region sequences also have been defined structurally by Chothia 30 as those residues that are not part of the conserved ⁇ -sheet framework, and thus are able to adapt different conformations (Chothia and Lesk, 1987, J. Mol. Biol. 196:901-917). Alternatively, IMGT or EU numbering may be used. These terminologies are well recognized in the art.
  • CDRs defined according to either the Kabat, Chothia, or IMGT designations are set forth in the Table 1 below.
  • Table 1 CDR Definitions 10
  • the binding molecule according to the invention may be an antibody or a fragment thereof comprising heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3 according to the IMGT numbering scheme, wherein the binding molecule specifically binds to CA19-9.
  • the binding molecule according to the present invention may be an antibody or a fragment thereof comprising HCDRs 1-3 and LCDRs 1-3 according to the IMGT numbering scheme, wherein at least one of the HCDRs 1-3 and/or LCDRs 1-3 is selected from: a) HCDR1 comprising an amino acid sequence according to SEQ ID NO: 1, b) HCDR2 comprising an amino acid sequence according to SEQ ID NO: 2, 20 c) HCDR3 comprising an amino acid sequence according to SEQ ID NO: 3, d) LCDR1 comprising an amino acid sequence according to SEQ ID NO: 4, e) LCDR2 comprising an amino acid sequence according to SEQ ID NO: 5, and f) LCDR3 comprisng an amino acid sequence according to SEQ ID NO: 6.
  • the binding molecule comprises one, two, three, four, five or six of the P129624PCT HCDRs 1-3 and/or LCDRs 1-3 selected from: a) HCDR1 comprising an amino acid sequence according to SEQ ID NO: 1, b) HCDR2 comprising an amino acid sequence according to SEQ ID NO: 2, c) HCDR3 comprising an amino acid sequence according to SEQ ID NO: 3, 5 d) LCDR1 comprising an amino acid sequence according to SEQ ID NO: 4, e) LCDR2 comprising an amino acid sequence according to SEQ ID NO: 5, and f) LCDR3 comprisng an amino acid sequence according to SEQ ID NO: 6.
  • HCDRs 1-3 and/or LCDRs 1-3 comprises one, two or three amino acid mutations relative to the recited sequences.
  • the binding molecule according to the present invention may be an antibody or a fragment thereof comprising heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3 according to the IMGT numbering scheme, wherein: a) HCDR1 comprises an amino acid sequence according to SEQ ID NO: 1, 15 b) HCDR2 comprises an amino acid sequence according to SEQ ID NO: 2, c) HCDR3 comprises an amino acid sequence according to SEQ ID NO: 3, d) LCDR1 comprises an amino acid sequence according to SEQ ID NO: 4, e) LCDR2 comprises an amino acid sequence according to SEQ ID NO: 5, f) LCDR3 comprises an amino acid sequence according to SEQ ID NO: 6; 20 optionally wherein one or more of the CDRs comprises one, two or three amino acid mutations relative to the recited
  • the binding molecule according to the present invention may be an antibody or a fragment thereof comprising heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3 according to the Kabat numbering 25 scheme, wherein: a) HCDR1 comprises an amino acid sequence according to SEQ ID NO: 15, b) HCDR2 comprises an amino acid sequence according to SEQ ID NO: 16, c) HCDR3 comprises an amino acid sequence according to SEQ ID NO: 17, d) LCDR1 comprises an amino acid sequence according to SEQ ID NO: 18, 30 e) LCDR2 comprises an amino acid sequence according to SEQ ID NO: 19, f) LCDR3 comprises an amino acid sequence according to SEQ ID NO: 6; P129624PCT optionally wherein one or more of the CDRs comprises one, two or three amino acid mutations relative to the recited sequences.
  • HCDR1 comprises an amino acid sequence according to SEQ ID NO: 15
  • b) HCDR2 comprises an amino acid sequence according
  • the binding molecule according to the present invention may be an antibody or a fragment thereof comprising heavy chain complementarity determining regions (HCDRs) 1-3 and light 5 chain complementarity determining regions (LCDRs) 1-3 according to the Chothia numbering scheme, wherein: a) HCDR1 comprises an amino acid sequence according to SEQ ID NO: 20, b) HCDR2 comprises an amino acid sequence according to SEQ ID NO: 21, c) HCDR3 comprises an amino acid sequence according to SEQ ID NO: 17, 10 d) LCDR1 comprises an amino acid sequence according to SEQ ID NO: 18, e) LCDR2 comprises an amino acid sequence according to SEQ ID NO: 19, f) LCDR3 comprises an amino acid sequence according to SEQ ID NO: 6; optionally wherein one or more of the CDRs comprises one, two or three amino acid mutations relative to the recited sequences.
  • HCDR1 comprises an amino acid sequence according to SEQ ID NO: 20
  • HCDR2 comprises an amino acid sequence according to SEQ ID NO: 21
  • one or more of the CDRs may comprise one, two or three amino acid mutations.
  • HCDR1 may comprise one, two or three amino acid mutations.
  • HCDR2 may comprise one, two or three amino acid mutations.
  • HCDR3 may comprise one, two or three amino acid 5 mutations.
  • LCDR1 may comprise one, two or three amino acid mutations.
  • LCDR2 may comprise one, two or three amino acid mutations.
  • LCDR3 may comprise one, two or three amino acid mutations.
  • a mutation in any of the CDRs described herein may encompass a 10 deletion of an amino acid, an insertion of an amino acid, or a substitution of an amino acid. It will also be understood that such a mutation may not prevent the binding molecule according to the invention from binding to CA19-9.
  • a binding molecule according to the invention comprising a mutation in one or more CDRs described herein may suitably maintain the capacity (e.g. affinity) to bind to CA19-9.
  • the mutation suitably 15 maintains the same capacity (e.g. affinity) to bind to CA19-9 as the parent binding molecule.
  • the term “parent binding molecule” in this context refers to the binding molecule without the mutation in question.
  • the binding molecule according to the present invention comprises a heavy chain variable region (VH) comprising an amino acid sequence according to SEQ ID 20 NO: 7, or a variant having at least 80% sequence identity thereto.
  • the binding molecule according to the present invention comprises a VH comprising an amino acid sequence according to SEQ ID NO: 7.
  • the binding molecule according to the present invention comprises a VH consisting of an amino acid sequence according to SEQ ID NO: 7. 25
  • the binding molecule according to the present invention comprises a light chain variable region (VL) comprising an amino acid sequence according to SEQ ID NO: 8, or a variant having at least 80% sequence identity thereto.
  • the binding molecule according to the present invention comprises a VL comprising an amino acid sequence according to SEQ ID NO: 8. In some embodiments, 30 the binding molecule according to the present invention comprises a VL consisting of an amino acid sequence according to SEQ ID NO: 8. P129624PCT In some embodiments, the binding molecule according to the present invention comprises a VH comprising an amino acid sequence according to SEQ ID NO: 7, or a variant having at least 80% sequence identity thereto, and a VL comprising an amino acid sequence according to SEQ ID NO: 8, or a variant having at least 80% identity thereto.
  • VH and/or VL having a percentage identity to the SEQ ID NO of any VH and/or VL defined herein may have an equivalent function to the VH and/or VL having the sequence set forth in the SEQ ID NO defined herein and may suitably maintain the capacity to bind to CA19-9, e.g. the same capacity as the VH and/or VL domain having the sequence set forth in the SEQ ID NO defined herein.
  • the VH has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 7.
  • the VL has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID 15 NO: 8.
  • the binding molecule comprises one or more immunoglobulin constant domains.
  • the immunoglobulin constant domains comprise a constant light chain domain (CL).
  • the immunoglobulin constant domains 20 comprise a constant heavy 1 (CH1) domain.
  • the immunoglobulin constant domains comprise a constant heavy 3 (CH3) domain.
  • the immunoglobulin constant domains comprise a constant heavy 2 (CH2) domain.
  • the immunoglobulin constant domains comprise a CH2 and a CH3 domain.
  • the CH2 and CH3 domains are considered to be an Fc (fragment 25 crystallisable) region.
  • the immunoglobulin constant domains comprise a CL, CH1, CH2 and CH3 domain.
  • the binding molecule comprises an Fc region. In some embodiments, the binding molecule comprises a modified Fc region. In some embodiments, the binding molecule further comprises a CL domain comprising an amino acid sequence according to SEQ ID NO: 22, or a variant having at least 70% identity 5 thereto. In some embodiments, the CL has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 22.
  • the binding molecule further comprises CH1, CH2 and CH3 domains comprising an amino acid sequence according to SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26 or SEQ ID NO: 27, or a variant having at least 70% identity thereto.
  • the CH1, CH2 and CH3 domain has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 15 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to any one of SEQ ID NOs: 23 to 27.
  • the binding molecule is an antibody.
  • the binding molecule is a monoclonal antibody.
  • the binding molecule is 5B1.
  • the binding molecule may be an antibody comprising a heavy chain(s) and a light chain(s).
  • the term “heavy chain” refers to a large protein subunit of an immunoglobulin. Heavy chains can be of any immunoglobulin isotype (for example IgG, IgE, IgM, IgD, IgA or IgY), subtype (for example lgG1, lgG2, lgG2a, lgG2b, lgG2c, lgG3, lgG4, lgA1 or lgA2) or allotype.
  • the term “light chain” refers to a small protein subunit of an immunoglobulin.
  • the antibody is an IgG1, preferably IgG1, lambda.
  • the antibody may be of any species (for example human, monkey, camel, llama, goat, sheep, rabbit, mouse, rat, mouse, hamster or chicken) or it may be a hybrid derived from more than 15 one species. It may be naturally occurring or it may be non-naturally occurring (i.e. an isolated antibody).
  • the antibody may be created by genetic engineering (for example a chimeric antibody, humanised antibody, camelised antibody, intrabody, bispecific antibody).
  • the binding molecule comprises a light chain comprising an amino acid sequence according to SEQ ID NO: 9, or a variant having at least 70% sequence identity 20 thereto.
  • the light chain has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 9.
  • the binding molecule comprises a heavy chain comprising an amino 25 acid sequence according to SEQ ID NO: 10, or a variant having at least 70% sequence identity thereto.
  • the heavy chain has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 10.
  • the binding molecule comprises a heavy chain comprising an amino acid sequence according to SEQ ID NO: 11, or a variant having at least 70% sequence identity thereto.
  • the heavy chain has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 5 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 11.
  • the binding molecule comprises a heavy chain comprising an amino acid sequence according to SEQ ID NO: 12, or a variant having at least 70% sequence identity thereto.
  • the heavy chain has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 12.
  • the binding molecule comprises a heavy chain comprising an amino acid sequence according to SEQ ID NO: 13, or a variant having at least 70% sequence identity 15 thereto.
  • the heavy chain has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 14.
  • the binding molecule comprises a light chain comprising SEQ ID NO: 9 or a variant having at least 70% sequence identity thereto and a heavy chain comprising SEQ ID NO: 10 or a variant having at least 70% sequence identity thereto.
  • the binding molecule comprises a light chain which consists of SEQ ID NO: 9 and a heavy chain which consists of SEQ ID NO: 10.
  • P129624PCT In some embodiments, the binding molecule comprises a light chain comprising SEQ ID NO: 9 or a variant having at least 70% sequence identity thereto and a heavy chain comprising SEQ ID NO: 11 or a variant having at least 70% sequence identity thereto. In some embodiments, the binding molecule comprises a light chain which consists of SEQ ID NO: 9 5 and a heavy chain which consists of SEQ ID NO: 11.
  • the binding molecule comprises a light chain which consists of SEQ ID NO: 9 15 and a heavy chain which consists of SEQ ID NO: 13. In some embodiments, the binding molecule comprises a light chain comprising SEQ ID NO: 9 or a variant having at least 70% sequence identity thereto and a heavy chain comprising SEQ ID NO: 14 or a variant having at least 70% sequence identity thereto. In some embodiments, the binding molecule comprises a light chain which consists of SEQ ID NO: 9 20 and a heavy chain which consists of SEQ ID NO: 14. P129624PCT P129624PCT Antibody The invention further provides an antibody comprising a modified Fc region, wherein the antibody specifically binds to CA19-9.
  • CA19-9 is considered the antigen of the antibodies according to the present invention.
  • the invention provides an antibody comprising heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3 according to the IMGT numbering scheme, wherein: 10 a) HCDR1 comprises an amino acid sequence according to SEQ ID NO: 1, b) HCDR2 comprises an amino acid sequence according to SEQ ID NO: 2, c) HCDR3 comprises an amino acid sequence according to SEQ ID NO: 3, d) LCDR1 comprises an amino acid sequence according to SEQ ID NO: 4, e) LCDR2 comprises an amino acid sequence according to SEQ ID NO: 5, 15 f) LCDR3 comprises an amino acid sequence according to SEQ ID NO: 6, optionally wherein one or more of the CDRs comprises one, two or three amino acid mutations relative to the recited sequences; and a modified Fc region.
  • HCDR1 comprises an amino acid sequence according to SEQ ID NO: 1
  • the invention provides an antibody comprising heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3 20 according to the Kabat numbering scheme, wherein: P129624PCT a) HCDR1 comprises an amino acid sequence according to SEQ ID NO: 15, b) HCDR2 comprises an amino acid sequence according to SEQ ID NO: 16, c) HCDR3 comprises an amino acid sequence according to SEQ ID NO: 17, d) LCDR1 comprises an amino acid sequence according to SEQ ID NO: 18, 5 e) LCDR2 comprises an amino acid sequence according to SEQ ID NO: 19, f) LCDR3 comprises an amino acid sequence according to SEQ ID NO: 6, optionally wherein one or more of the CDRs comprises one, two or three amino acid mutations relative to the recited sequences; and a modified Fc region.
  • HCDR1 comprises an amino acid sequence according to SEQ ID NO: 15
  • b) HCDR2 comprises an amino acid sequence according to SEQ ID NO:
  • the invention provides an antibody comprising heavy chain complementarity determining 10 regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3 according to the Chothia numbering scheme, wherein: a) HCDR1 comprises an amino acid sequence according to SEQ ID NO: 20, b) HCDR2 comprises an amino acid sequence according to SEQ ID NO: 21, c) HCDR3 comprises an amino acid sequence according to SEQ ID NO: 17, 15 d) LCDR1 comprises an amino acid sequence according to SEQ ID NO: 18, e) LCDR2 comprises an amino acid sequence according to SEQ ID NO: 19, f) LCDR3 comprises an amino acid sequence according to SEQ ID NO: 6, optionally wherein one or more of the CDRs comprises one, two or three amino acid mutations relative to the recited sequences; and a modified Fc region.
  • HCDR1 comprises an amino acid sequence according to SEQ ID NO: 20
  • HCDR2 comprises an amino acid sequence according to SEQ ID NO: 21
  • HCDR3 comprises
  • one or more of the CDRs may comprise one, two or three amino acid mutations.
  • HCDR1 may comprise one, two or three amino acid mutations.
  • HCDR2 may comprise one, two or three amino acid mutations.
  • HCDR3 may comprise one, two or three amino acid mutations.
  • LCDR1 may comprise one, two or three amino acid 25 mutations.
  • LCDR2 may comprise one, two or three amino acid mutations.
  • LCDR3 may comprise one, two or three amino acid mutations.
  • a mutation in any of the CDRs described herein may encompass a deletion of an amino acid, an insertion of an amino acid, or a substitution of an amino acid. It 30 will also be understood that such a mutation may not prevent the antibody according to the invention from binding to CA19-9.
  • an antibody according to the invention P129624PCT comprising a mutation in one or more CDRs described herein may suitably maintain the capacity (e.g. affinity) to bind to CA19-9.
  • the mutation suitably maintains the same capacity (e.g. affinity) to bind to CA19-9 as the parent antibody.
  • the term “parent antibody” in this context refers to the antibody without the mutation in question.
  • antibody refers to a glycoprotein comprising at least two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds, and includes any molecule comprising an antigen binding portion thereof.
  • antibody includes monoclonal antibodies and fragments or derivatives of antibodies, including, without limitation, human antibodies, humanized antibodies, chimeric antibodies, single chain antibodies, e.g., scFvs and antigen- 10 binding antibody fragments such as Fab and Fab' fragments and also includes all recombinant forms of antibodies, e.g., antibodies expressed in prokaryotes, unglycosylated antibodies, and any antigen-binding antibody fragments and derivatives as described herein.
  • the antibody according to the present invention comprises a heavy chain variable region (VH) comprising an amino acid sequence according to SEQ ID NO: 7, 15 or a variant having at least 80% sequence identity thereto.
  • the antibody according to the present invention comprises a VH comprising an amino acid sequence according to SEQ ID NO: 7.
  • the antibody according to the present invention comprises a VH consisting of an amino acid sequence according to SEQ ID NO: 7. 20
  • the antibody according to the present invention comprises a light chain variable region (VL) comprising an amino acid sequence according to SEQ ID NO: 8, or a variant having at least 80% sequence identity thereto.
  • the antibody according to the present invention comprises a VL comprising an amino acid sequence according to SEQ ID NO: 8.
  • the 25 antibody according to the present invention comprises a VL consisting of an amino acid sequence according to SEQ ID NO: 8.
  • the antibody according to the present invention comprises a VH comprising an amino acid sequence according to SEQ ID NO: 7, or a variant having at least 80% sequence identity thereto, and a VL comprising an amino acid sequence according to 30 SEQ ID NO: 8, or a variant having at least 80% identity thereto.
  • any VH and/or VL defined herein may have an equivalent function to the VH and/or VL having the sequence set forth in the SEQ ID NO defined herein and may suitably maintain the capacity to bind to CA19-9, e.g. the same capacity as the VH and/or VL domain having the sequence set forth in the SEQ ID NO defined herein.
  • the VH has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 7.
  • the VL has at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID 10 NO: 8.
  • the antibody comprises a light chain comprising an amino acid sequence according to SEQ ID NO: 9, or a variant having at least 70% sequence identity thereto.
  • the light chain has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 15 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 9.
  • the antibody according to the present invention comprises a modified Fc region.
  • the antibody comprises a heavy chain comprising an amino acid sequence according to SEQ ID NO: 11, or a variant having at least 70% sequence identity 20 thereto.
  • the heavy chain has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 11.
  • the antibody comprises a heavy chain comprising an amino acid 25 sequence according to SEQ ID NO: 12, or a variant having at least 70% sequence identity thereto.
  • the heavy chain has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 12.
  • the antibody comprises a heavy chain comprising an amino acid sequence according to SEQ ID NO: 13, or a variant having at least 70% sequence identity thereto.
  • the heavy chain has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 5 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 13.
  • the antibody comprises a heavy chain comprising an amino acid sequence according to SEQ ID NO: 14, or a variant having at least 70% sequence identity thereto.
  • the heavy chain has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 14.
  • the antibody comprises a light chain comprising SEQ ID NO: 9 or a variant having at least 70% sequence identity thereto and a heavy chain comprising SEQ ID 15 NO: 11 or a variant having at least 70% sequence identity thereto.
  • the antibody comprises a light chain which consists of SEQ ID NO: 9 and a heavy chain which consists of SEQ ID NO: 11. In some embodiments, the antibody comprises a light chain comprising SEQ ID NO: 9 or a variant having at least 70% sequence identity thereto and a heavy chain comprising SEQ ID 20 NO: 12 or a variant having at least 70% sequence identity thereto. In some embodiments, the antibody comprises a light chain which consists of SEQ ID NO: 9 and a heavy chain which consists of SEQ ID NO: 12. In some embodiments, the antibody comprises a light chain comprising SEQ ID NO: 9 or a variant having at least 70% sequence identity thereto and a heavy chain comprising SEQ ID 25 NO: 13 or a variant having at least 70% sequence identity thereto.
  • the antibody comprises a light chain which consists of SEQ ID NO: 9 and a heavy chain which consists of SEQ ID NO: 13. In some embodiments, the antibody comprises a light chain comprising SEQ ID NO: 9 or a variant having at least 70% sequence identity thereto and a heavy chain comprising SEQ ID 30 NO: 14 or a variant having at least 70% sequence identity thereto. In some embodiments, the antibody comprises a light chain which consists of SEQ ID NO: 9 and a heavy chain which P129624PCT consists of SEQ ID NO: 14. Modified Fc region In some embodiments, the binding molecule, e.g. via the Fc region, binds to one or more or all of the Fc receptors.
  • the Fc receptors comprise one or more or 5 all of Fc ⁇ RI (CD64), Fc ⁇ RIIa (CD32A), Fc ⁇ RIIb (CD32B), Fc ⁇ RIII (CD16), C1q and FcRn.
  • the binding molecule e.g. via the Fc region, binds to Fc ⁇ RI.
  • the Fc region may interact with Fc receptors presented on the surface of a cell and/or may interact with proteins of the complement system.
  • the Fc receptors may be Fc gamma receptors, e.g. Fc ⁇ RI.
  • the proteins of the complement system may include C1q.
  • the Fc region of the binding molecule is silenced to reduce, negate or abolish one or more Fc receptor binding and/or functionalities.
  • the Fc region of the binding molecule is modified to negate one or more Fc receptor functionalities.
  • the Fc region of the binding molecule is silenced in respect of one or more or all of Fc ⁇ RI (CD64), Fc ⁇ RIIa (CD32A), Fc ⁇ RIIb (CD32B), Fc ⁇ RIII (CD16) and C1q 15 functionality.
  • the Fc region of the binding molecule as defined herein is a modified Fc region.
  • the Fc region of the binding molecule according to the invention may not be capable of binding to immune cells and/or recruiting immune cells.
  • the antibody according to the invention comprises a modified Fc region. That is, the Fc region of the antibody of the invention molecule is silenced to reduce, negate or abolish one or more Fc receptor binding and/or functionalities. In some embodiments, the Fc region of the antibody of the invention is modified to negate one or more Fc receptor functionalities. In some embodiments, the Fc region of the antibody of the invention is silenced in respect of one or 25 more or all of Fc ⁇ RI (CD64), Fc ⁇ RIIa (CD32A), Fc ⁇ RIIb (CD32B), Fc ⁇ RIII (CD16) and C1q functionality.
  • the Fc region of the antibody according to the invention may not be capable of binding to immune cells and/or recruiting immune cells.
  • the binding of the modified Fc region to Fc ⁇ RI (CD64), Fc ⁇ RIIa (CD32A), Fc ⁇ RIIb (CD32B), P129624PCT Fc ⁇ RIII (CD16), C1q and FcRn may be reduced compared to a wild-type Fc region.
  • the binding of the modified Fc region to Fc ⁇ RI may be reduced compared to a wild-type Fc region.
  • the binding molecule comprises an Fc region comprising an amino acid substitution at position 234, or an amino acid substitution at position 235, or an amino acid 10 substitution at position 236, or an amino acid substitution at positions 234 and 235, or an amino acid substitution at positions 234, 235 and 236, according to the EU numbering scheme.
  • the Fc region may comprise further amino acid substitutions.
  • the substitution(s) may be relative to a wild-type Fc region.
  • the antibody comprises an Fc region comprising an amino acid substitution 15 at position 234, or an amino acid substitution at position 235, or an amino acid substitution at position 236, or an amino acid substitution at positions 234 and 235, or an amino acid substitution at positions 234, 235 and 236, according to the EU numbering scheme.
  • the Fc region may comprise further amino acid substitutions.
  • the substitution(s) may be relative to a wild-type Fc region.
  • a wild-type Fc region may comprise the amino acid sequence according to SEQ ID NO: 28.
  • the binding molecule comprises an Fc region comprising amino acid substitutions at positions 234 and 235, according to the EU numbering scheme.
  • the binding molecule comprises an Fc region comprising amino acid P129624PCT substitutions at positions 234, 235 and 236, according to the EU numbering scheme.
  • the Fc region of the binding molecule comprises a silencing modification selected from the STR mutation, the LALA mutation, the LALA- ⁇ A mutation and the LALA-KA mutation as defined herein.
  • the antibody comprises an Fc region comprising amino acid substitutions at positions 234 and 235, according to the EU numbering scheme.
  • the antibody comprises an Fc region comprising amino acid substitutions at positions 234, 235 and 236, according to the EU numbering scheme.
  • the Fc region of the antibody comprises a silencing modification 10 selected from the STR mutation, the LALA mutation, the LALA- ⁇ A mutation and the LALA-KA mutation as defined herein.
  • the amino acid substitution(s) may be selected from the group consisting of: a) L234A, L235A, A327G, A330S, and P331S (LALA- ⁇ A); b) L234A, L235A, and K322A (LALA-KA); 15 c) L234S, L235T, and G236R (STR); and d) L234A and L235A (LALA).
  • the Fc region of the binding molecule or antibody comprises the LALA Mutation: L234A, L235A by EU numbering (or L247A, L248A by Kabat numbering; or L4A, L5A by IMGT(CH2) numbering).
  • the Fc region of the binding molecule or antibody comprises the LALA delta-A mutation: L234A, L235A, A327G, A330S, P331S by EU numbering (or L247A, L248A, A346G, A349S, P350S by Kabat numbering; or L4A, L5A, A97G, A100S, P101S by IMGT(CH2) numbering).
  • the Fc region of the binding molecule or antibody comprises the LALA-KA 25 mutation: L234A, L235A, K322A by EU numbering (or L247A, L248A, K341A by Kabat numbering; or L4A, L5A, K92A by IMGT(CH2) numbering)
  • the Fc region of the binding molecule or antibody comprises the amino acid residue A at position 234 and A at position 235 by EU numbering.
  • P129624PCT In an embodiment, the Fc region of the binding molecule or antibody comprises the amino acid residue A at position 234, A at position 235, G at position 327, S at position 330 and S at position 331 by EU numbering.
  • the Fc region of the binding molecule or antibody comprises the amino 5 acid residue A at position 234, A at position 235 and A at position 322 by EU numbering.
  • the Fc region of the binding molecule or antibody comprises the STR Mutation: L234S, L235T, G236R by EU numbering (or L247S, L248T, G249R by Kabat numbering; or L4S, L5T, G6R by IMGT(CH2) numbering).
  • the Fc region of the binding molecule or antibody comprises the amino 10 acid residue S at position 234, T at position 235 and R at position 236 by EU numbering.
  • the Fc region may comprise amino acid substitution(s) selected from the group consisting of: L234A/L235A/G236R, L234A/L235S/G236R , L234A/L235T/G236R, L234D/L235H/G236R, L234D/L235K/G236R, L234D/L235Q/G236R, L234D/L235S/G236R, L234D/L235T/G236R, L234E/L235D/G236R, L234E/L235H/G236R, L234E/L235I/G236R, L234E/L235V/G236R, 15 L234G/L235H/G236R, L234G/L235Q/G236R, L234G/L235S/G236R, L234H/L235I/G236R, L234H/L235S
  • Binding molecule-payload construct The present invention provides a binding molecule-payload conjugate (BPC) comprising a binding molecule as defined herein and one or more payload moieties wherein the one or 30 more payload moieties is covalently linked to the binding molecule, optionally via one or more linkers. Where more than one payload moiety is covalently linked to the binding molecule, P129624PCT optionally via one or more linkers, the payload moieties may be the same or different, and/or the linkers may be the same or different. It will be understood that the term “payload” may be interchangeable with “cargo”.
  • BPC is analogous to the term “ADC”, as in 5 “antibody-drug conjugate”, except that the term “binding molecule” as used herein encompasses antibodies and fragments thereof, and is not limited to full-length antibodies per se.
  • ADCs are a class of targeted therapeutics that can improve the selectivity and the cytotoxic activity of cancer drugs.
  • the ADC may become internalised and trafficked to intracellular compartments (e.g. a lysosome) where the payload is released from the ADC.
  • the payload is a drug
  • the release of the drug from the ADC may allow the drug to exert its effect on the cell.
  • the payload may be released from the ADC by proteolysis of a cleavable linker (if present) or by degradation of the antibody of the ADC.
  • the payload moiety is selected from a drug, a detectable marker, a radioisotope (which may be a radiotherapeutic agent and/or a radioimaging agent), a fluorescent agent, a luminescent agent, a colored agent, an enzyme, polyethylene glycol, a nuclide, a nucleic acid, a small molecule toxin, a polypeptide having binding activity, a protein, a receptor, a ligand, another active agent that inhibits tumor cell growth, promotes tumor cell 20 apoptosis or necrosis.
  • the payload moiety is a drug.
  • the payload moiety is a cytotoxic drug, an immune modulator, or a STING inhibitor
  • the cytotoxic drug may be a tubulin inhibitor, a DNA damaging agent, a TOPO1 inhibitor, an auristatin, a maytansinoid, or a calicheamicin.
  • the cytotoxic drug is a topoisomerase inhibitor, which may be a type I topoisomerase (TOPO1) inhibitor or a type II topoisomerase (TOPO2) inhibitor.
  • the TOPO1 inhibitor is a camptothecin or an exatecan.
  • Camptothecins may include topotecan, irinotecan, and belotecan.
  • the cytotoxic drug is a tubulin inhibitor (also known as a microtubule 30 inhibitor).
  • the cytotoxic drug is an auristatin.
  • P129624PCT the auristatin is monomethyl auristatin E.
  • the auristatin is monomethyl auristatin F.
  • the cytotoxic drug is a maytansinoid.
  • a first payload moiety is a topoisomerase inhibitor 5 (as defined and exemplified above) and a second payload moiety is a tubulin inhibitor (as defined and exemplified above).
  • a cytotoxic drug, chemotherapeutic drug, or chemotherapeutic entity may refer to a drug or molecule that: • is destructive to a cell; 10 • induces apoptosis in a cell; • inhibits or prevents the function of a cell; • inhibits or prevents a cell from proliferating; and/or • reduces the viability of a cell.
  • the invention comprises a BPC compound as defined herein. In other 15 embodiments, the invention comprises a tautomer of the BPC as defined herein. In other embodiments, the invention comprises a mesomer of the BPC as defined herein. In other embodiments, the invention comprises a racemate of the BPC as defined herein. In other embodiments, the invention comprises an enantiomer of the BPC as defined herein. In other embodiments, the invention comprises a diastereoisomer of the BPC as defined herein. In 20 other embodiments, the invention comprises a composition which is a mixture of the BPCs as defined herein. In other embodiments, the invention comprises a pharmaceutically acceptable salt of the BPC as defined herein.
  • Linker and payload moieties typically, in the BPCs of the present invention, one or more payload moieties is covalently 25 linked to the binding molecule via a linker to form the BPC.
  • the moiety linked to the binding molecule is generally referred to herein as the “linker-payload moiety”.
  • linker-payload moiety typically, when the BPC is administered to the subject, the linker-payload moiety is released by breakdown of the BPC in vivo. The linker-payload moiety then typically breaks down to release the payload in vivo.
  • the drug-antibody ratio is the number of linker-payload molecules attached to each P129624PCT binding molecule (it can be understood that the term “drug-antibody ratio” applies equally whether the binding molecule is an antibody or a fragment thereof).
  • the drug-antibody ratio (DAR) of the BPCs according to the invention may vary.
  • the terms “drug-antibody ratio”, “DAR” and “connection number” are 5 synonymous.
  • the DAR is an integer from 1 to 16.
  • the DAR is an integer of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16.
  • the DAR is an integer from 4 to 12.
  • DAR is an integer from 6 to 10.
  • the DAR is an integer from 7 to 9. In some embodiments, the DAR is 6. 10 In some embodiments, the DAR is 7. In some embodiments, the DAR is 8. In some embodiments, the DAR is 9. In some embodiments, the DAR is 10.
  • the linker unit comprises a first conjugation moiety for coupling with the binding molecule. In some embodiments, the linker unit comprises a second conjugation moiety for coupling with 15 the payload moiety. In some embodiments, the linker unit comprises a peptide linker. In some embodiments, the linker unit consists essentially of a peptide linker. In some embodiments, the linker unit consists of a peptide linker.
  • the peptide linker comprises, essentially of or consist of 1 to 10 amino acid residues.
  • the peptide linker may comprise, consist essentially of or 20 consist of 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid residues.
  • the peptide linker comprises, consists essentially of or consists of 1 to 6 amino acid residues.
  • the peptide linker comprises, consists essentially of or consists of 1 to 5 amino acid residues.
  • the peptide linker comprises, consists essentially of or consists of 2 amino acid residues.
  • the peptide linker comprises, 25 consists essentially of or consists of 3 amino acid residues.
  • the peptide linker comprises, consists essentially of or consists of 4 amino acid residues. In some embodiments, the peptide linker comprises, consists essentially of or consists of 5 amino acid residues. In some embodiments, the linker comprises a peptide linker selected from AA 1 , AA 1 -Gly, Val-30 Cit, Val-Ala, Val-AA 1 , Val-AA 1 -Gly, AA 1 -Ala-Asn, Ala-Ala-Ala, Ala-Ala-Asn and Gly-Gly-Phe- Gly.
  • the amino acid residue represented by AA 1 is selected from 5 where position 1 is connected, optionally via a further conjugation moiety, to the portion of the molecule bearing the binding compound and position 2 is connected, optionally via a further conjugation moiety, to the portion of the molecule bearing the payload compound.
  • the first conjugation moiety comprises a pyrimidine-sulfone moiety.
  • the pyrimidine-sulfone moiety is linked to the peptide moiety via a C1-10 10 alkynoyl group.
  • the linker unit comprises a peptide linker comprising the peptide sequence Gly-Gly-Phe-Gly.
  • the first moiety comprises a maleimide moiety which is capable of adding to a sulfhydryl moiety on the binding compound such that the conjugate comprises a 15 sulfur-linked succinimidyl group.
  • the maleimide moiety is linked to the peptide moiety via a C1-10 alkanoyl group.
  • connection number q is an integer of 1, 2, 3, 4, 5, 5 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16. In some embodiments of formula (III-D), the connection number q is an integer from 4 to 12. In some embodiments of formula (III-D), the connection number q is an integer from 6 to 10. In some embodiments of formula (III-D), the connection number q is an integer from 7 to 9. In some embodiments of formula (III-D), the connection number q is 6. In some embodiments, the connection number q is 7. In some embodiments 10 of formula (III-D), the connection number q is 8.
  • connection number q is 9. In some embodiments of formula (III-D), the connection number q is 10. In some embodiments, wherein W is -(C(R wa )(R wb )) wn -, Y is -(OCH 2 CH 2 ) yn -O yp -, and Z is -(C(R za )(R zb )) zn ; 15 wherein wn is 1, 2, 3 or 6, and 1 methylene unit of W is independently replaced by -Cyr-, -N(R wx )C(O)-, -C(O)N(R wx )- or - C(O)-; wherein yn is 0, 4 or 8, and yp is 0 or 1; P129624PCT wherein zn is 1, 2 or 3, and 1 methylene unit of Z is independently replaced by -Cyr-, -N(R zx )
  • -L b - is selected from the group consisting of: 10 R L1 R L2
  • -L c - is , wherein R L1 and R L2 are each independently selected from the group consisting of: hydrogen, halogen, -OH and a C 1-6 aliphatic group.
  • -L a - is , 15 wherein W is -(C(R wa )(R wb )) wn -, Y is -(OCH 2 CH 2 ) yn -O yp -, and Z is -(C(R za )(R zb )) zn ; wherein wn is 1, 2, 3 or 6, and 1 methylene unit of W is independently replaced by -Cyr-, -N(R wx )C(O)-, -C(O)N(R wx )- or - C(O)-; P129624PCT wherein yn is 0, 4 or 8, and yp is 0 or 1; wherein zn is 1, 2 or 3, and 1 methylene unit of Z is independently replaced by -Cyr-, -N(R zx )C(O)-, -C(O)N(R zx )- or -C(
  • R L1 and R L2 are each independently selected from the group consisting of: hydrogen, halogen, -OH and a C 1-6 aliphatic group. In some embodiments, In some embodiments, 15 In some embodiments, In some embodiments, In some embodiments, In some embodiments, the payload-linker moiety comprises the following group of structures: P129624PCT III-A-16 III-A-17 In some embodiments, the payload moiety of the conjugate has the following structure: . In some embodiments, the payload-linker moiety of the conjugate has the following structure: P129624PCT In some embodiments, the conjugate is selected from the group consisting of the following structural formulae: 5
  • the conjugate has the structure of Formula (IVA):
  • connection number q represents a connection number
  • n is selected from the group consisting of integers from 1 to 16
  • BM is a binding molecule as defined herein. 5
  • the connection number q is an integer of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16.
  • the connection number q is an integer from 4 to 12.
  • the connection number q is an integer from 6 to 10.
  • the connection number q is an integer from 7 to 9.
  • the connection 10 number q is 6.
  • connection number q is 7. In some embodiments of formula (IVA), the connection number q is 8. In some embodiments of formula (IVA), the connection number q is 9. In some embodiments of formula (IVA), the connection number q is 10. In some embodiments, the payload or payload-linker moiety comprises a structure shown as 15 formula (II-A):
  • X 1 is saturated C, and X 1 is substituted with R n ;
  • ring A is selected from the group consisting of: 3-10 membered saturated or partially unsaturated heterocyclyl and 3-10 membered saturated or partially unsaturated carbocyclyl, 5 wherein ring A is substituted with 0 or at least 1 substituent R 1a ; when ring A is 3-10 membered saturated or partially unsaturated carbocyclyl, ring A is substituted with p L 2 , and L 2 is not R n ; or, when ring A is 3-10 membered saturated or partially unsaturated heterocyclyl, ring A is substituted with p L 2 ; 10 L 2 is -R 2 -L 3 -, and R 2 is used for direct or indirect linking of a ligand; L 3 is -(C(R 3a )(R 3b )) m -, wherein when L 3 comprises a methylene unit, 0 or at least 1
  • the payload or payload-linker moiety comprises a structure shown as 10 formula (II-C x ): , wherein, L is -L a -L b -L c -; -L a - is selected from the group consisting of: 15 wherein W is -(C(R wa )(R wb )) wn -, Y is -(OCH 2 CH 2 ) yn -O yp -, and Z is -(C(R za )(R zb )) zn ; wherein wn is selected from the group consisting of integers ⁇ 0, and 0 or at least 1 methylene unit of W is independently replaced by -Cyr-, -N(R wx )C(O)-, - P129624PCT C(O)N(R wx )-, -C(O)-, -OC(O)-, -C(O
  • the BPC comprises a structure shown as formula (II-D x ): 5 , wherein BM is the binding molecule, and q is a connection number which is an integer from 1 to 16.
  • the connection number q is an integer of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16.
  • the 10 connection number q is an integer from 4 to 12.
  • the connection number q is an integer from 6 to 10.
  • the connection number q is an integer from 7 to 9.
  • the connection number q is 6.
  • connection number q is 7. In some embodiments of formula (II-Dx), the connection number q is 8. In some 15 embodiments of formula (II-Dx), the connection number q is 9. In some embodiments of formula (II-Dx), the connection number q is 10.
  • -L a- is selected from the group consisting of: P129624PCT wherein W is -(C(R wa )(R wb )) wn -, Y is -(OCH 2 CH 2 ) yn -O yp -, and Z is -(C(R za )(R zb )) zn ; wherein wn is selected from the group consisting of integers from 2 to 6, and 0 or 1 methylene unit of W is independently replaced by -Cyr-, -N(R wx )C(O)-, -C(O)N(R wx )-, - C(O)-, -NR wx - or -O-; 5 wherein yn is selected from the group consisting of integers from 0 to 12, and yp is 0 or 1; wherein zn is selected from the group consisting of integers from 0 to 10, and 0
  • W is -(C(R wa )(R wb )) wn -
  • Y is -(OCH 2 CH 2 ) yn -O yp -
  • Z is -(C(R za )(R zb )) zn ; wherein wn is 1, 2, 3 or 6, and 1 methylene unit of W is independently replaced by -Cyr-, -N(R wx )C(O)-, -C(O)N(R wx )- or - 10 C(O)-; wherein yn is 0, 4 or 8, and yp is 0 or 1; wherein zn is 1, 2 or 3, and 1 methylene unit of Z is independently replaced by -Cyr-, -N(R zx )C(O)-, -C(O)N(R zx )- or -C(O)-; -Cyr- is 3-10 membered
  • -L b - represents a peptide residue consisting of 2 to 4 amino acids
  • 20 the peptide residue of -L b - is a peptide residue formed of amino acids selected from the group consisting of: phenylalanine, glycine, alanine, valine, citrulline and lysine
  • -L c - is selected from the group consisting of: P129624PCT wherein R L1 and R L2 are each independently selected from the group consisting of: hydrogen, halogen, -OH and a C 1-6 aliphatic group.
  • -L a - is 5 , wherein W is -(C(R wa )(R wb )) wn -, Y is -(OCH 2 CH 2 ) yn -O yp -, and Z is -(C(R za )(R zb )) zn ; wherein wn is 1, 2, 3 or 6, and 1 methylene unit of W is independently replaced by -Cyr-, -N(R wx )C(O)-, -C(O)N(R wx )- or - C(O)-; 10 wherein yn is 0, 4 or 8, and yp is 0 or 1; wherein zn is 1, 2 or 3, and 1 methylene unit of Z is independently replaced by -Cyr-, -N(R zx )C(O)-, -C(O)N(R zx )- or -C(O)-;
  • -L a - is , wherein W is -(C(R wa )(R wb )) wn -, Y is -(OCH 2 CH 2 ) yn -O yp -, and Z is -(C(R za )(R zb )) zn ; wherein wn is 1, 2, 3 or 6, and 10 1 methylene unit of W is independently replaced by -Cyr-, -N(R wx )C(O)-, -C(O)N(R wx )- or - C(O)-; wherein yn is 0, 4 or 8, and yp is 0 or 1; wherein zn is 1, 2 or 3, and 1 methylene unit of Z is independently replaced by -Cyr-, -N(R zx )C(O)-, -C(O)N(R zx )- or -C(O)-; 15
  • -L c - is wherein R L1 and R L2 are each independently selected from the group consisting of: hydrogen, halogen, -OH and a C 1-6 aliphatic group. 5
  • ring A is selected from the group consisting of: 3-10 membered 10 saturated heterocyclyl and 3-10 membered saturated carbocyclyl. In some embodiments, ring A is 3-10 membered saturated carbocyclyl. In some embodiments, ring A is 3-6 membered saturated carbocyclyl. In some embodiments, ring A is 4 membered saturated carbocyclyl. In some embodiments, ring A is 6 membered saturated carbocyclyl.
  • ring A is 3-10 membered saturated heterocyclyl. In some embodiments, ring A is 3-6 membered saturated heterocyclyl. In some embodiments, ring A is 4 membered saturated heterocyclyl. In some embodiments, ring A is substituted with 0 substituent R 1a .
  • the linker-payload moiety comprises a structure shown as formula (II- P129624PCT Ax): wherein, when ring A is 3-10 membered saturated or partially unsaturated carbocyclyl, ring A is substituted with p L 2 , and L 2 is not R n ; 5 or, when ring A is 3-10 membered saturated or partially unsaturated heterocyclyl, ring A is substituted with p L 2 . In some embodiments, ring A is substituted with 1 L 2 . In some embodiments, m is 0, and L 3 is a covalent bond. In some embodiments, m is 2, and L 3 is -(C(R 3a )(R 3b )) 2 -.
  • 1 methylene unit 15 of L 1 is replaced by -C(O)-.
  • R 2 is selected from the group consisting of: -O-, -(R 2a )N- and -S-. In some embodiments, R 2 is -O-. In some embodiments, R 2 is -(R 2a )N-. In some embodiments, R 2a is hydrogen. In some embodiments, wherein R 2 is -HN-. P129624PCT In some embodiments, R 1a is hydrogen. In some embodiments, R 3a and R 3b are each independently hydrogen. In some embodiments, R 4 is hydrogen. In some embodiments, R 6 is hydrogen.
  • R, R a and R b are each independently hydrogen.
  • the payload or payload-linker moiety comprises the following group of structures: II-A-9 II-A-10 II-A-11 II-A-12 In some embodiments, the payload or payload-linker moiety comprises the structure P129624PCT In some embodiments, the payload or payload-linker moiety has the structure In some embodiments, the payload-linker moiety has the structure 5 .
  • the conjugate moiety has the structure of formula (IVB) P129624PCT or a pharmaceutically acceptable salt thereof, wherein, q represents a connection number, and q is selected from the group consisting of integers from 1 to 16, and BM is a binding molecule as defined herein.
  • the connection number q is an integer of 1, 2, 3, 4, 5, 5 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16.
  • the connection number q is an integer from 4 to 12.
  • the connection number q is an integer from 6 to 10.
  • the connection number q is an integer from 7 to 9.
  • connection number q is 6. In some embodiments of formula (IVB), the connection number q is 7. In some 10 embodiments of formula (IVB), the connection number q is 8. In some embodiments of formula (IVB), the connection number q is 9. In some embodiments of formula (IVB), the connection number q is 10. 15 or a pharmaceutically acceptable salt thereof, wherein, q represents a connection number, and n is selected from the group consisting of integers from 1 to 16, and BM is a binding molecule as defined herein.
  • the payload-linker moiety of the compound of formula (VA) is also known as deruxtecan.
  • connection number q is an integer of 1, 2, 3, 4, 5, 20 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16. In some embodiments of formula (VA), the connection number q is an integer from 4 to 12. In some embodiments of formula (VA), the connection number q is an integer from 6 to 10. In some embodiments of formula (VA), the connection P129624PCT number q is an integer from 7 to 9. In some embodiments of formula (VA), the connection number q is 6. In some embodiments of formula (VA), the connection number q is 7. In some embodiments of formula (VA), the connection number q is 8. In some embodiments of formula (VA), the connection number q is 9. In some embodiments of formula (VA), the connection 5 number q is 10.
  • connection number q is an integer of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16. In some embodiments of formula (I), the connection number q is an integer from 4 to 12. In some embodiments of formula (I), the connection number q is an integer from 6 to 10. In some embodiments of formula (I), the connection number q is an integer from 7 to 9. In some embodiments of formula (I), the connection number 15 q is 6. In some embodiments of formula (I), the connection number q is 7. In some embodiments of formula (I), the connection number q is 8. In some embodiments of formula (I), the connection number q is 9. In some embodiments of formula (I), the connection number q is 10.
  • Z is selected from a direct bond, a carbon-carbon triple bond, and a carbon-carbon double bond. In some embodiments, Z is a carbon-carbon triple bond.
  • Rx is H or methyl. In some embodiments, Rx is H. 5
  • Ry is H or methyl. In some embodiments, Ry is H. In some embodiments, m is 2, 3 or 4. In some embodiments, m is 3. In some embodiments, L 2 is absent.
  • L3 is selected from AA 1 , AA 1 -Gly, Val-Cit, Val-Ala, Val-AA 1 , Val-AA 1 -Gly, AA 1 -Ala-Asn, Ala-Ala-Ala, Ala-Ala-Asn and Gly-Gly-Phe-Gly.
  • L 3 is 10 selected from AA 1 , AA 1 -Gly, Val-Cit, Val-AA 1 -Gly, AA 1 -Ala-Asn and Gly-Gly-Phe-Gly.
  • L 3 is Val-AA 1 -Gly.
  • the amino acid residue represented by AA 1 is selected from In some embodiments, the amino acid residue represented by AA 1 is , 15 In some embodiments, L 4 is selected from H , and . P129624PCT 1 N 2 In some embodiments, L4 is H . In some embodiments, W is O. In some embodiments, X is -(CH 2 ) n1 -. In some embodiments, n1 is 2, 3 or 4. In some embodiments, n1 is 3. 5 In some embodiments, the structure is: wherein position 1 is attached to the connecting atom on the binding molecule and position 2 is attached to W. ts, the structural fragment represented wherein position 1 is attached to L4.
  • the linker-payload moiety has the structure: 5
  • the conjugate is selected from the group consisting of: P129624PCT wherein, BM is the binding compound as defined herein q represents a connection number, and is selected from the group consisting of integers from 1 to 16, and BM is a binding molecule as defined herein. 5
  • the conjugate has the structure of formula (IVC): wherein, q represents a connection number, and is selected from the group consisting of integers from 1 to 16, and BM is a binding molecule as defined herein.
  • connection number q is an integer of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16. In some embodiments of formula (IVC), the connection number q is an integer from 4 to 12. In some embodiments of formula (IVC), the connection number q is an integer from 6 to 10. In some embodiments of formula (IVC), the connection number q is an integer from 7 to 9. In some embodiments of formula (IVC), the connection 15 number q is 6. In some embodiments of formula (IVC), the connection number q is 7. In some embodiments of formula (IVC), the connection number q is 8. In some embodiments of formula (IVC), the connection number q is 9. In some embodiments of formula (IVC), the connection number q is 10.
  • the invention provides a method for producing a BPC according to the invention, comprising contacting a binding molecule as defined herein with a suitable linker-payload compound.
  • the invention provides a method for producing a BPC according to the invention, comprising 5 contacting an antibody according to the invention with a suitable linker-payload compound.
  • the linker-payload compound may be of the general formula (II-Fx): , wherein, L x is L ax -L b -L c -; L ax - is selected from the group consisting of: 10 wherein R hal is iodine or bromine; wherein W is -(C(R wa )(R wb )) wn -, Y is -(OCH 2 CH 2 ) yn -O yp -, and Z is -(C(R za )(R zb )) zn ; wherein wn is selected from the group consisting of integers ⁇ 0, and P129624PCT 0 or at least 1 methylene unit of W is independently replaced by -Cyr-, -N(R wx )C(O)-, - C(O)N(R wx )-, -C(O)-, -OC(O)-, -
  • the linker-payload compound may be of the general formula (III-F): 5 , wherein, L x is L ax -L b -L c -; L ax - is selected from the group consisting of: wherein R hal is iodine or bromine; 10 wherein W is -(C(R wa )(R wb )) wn -, Y is -(OCH 2 CH 2 ) yn -O yp , and Z is -(C(R za )(R zb )) zn ; wherein wn is selected from the group consisting of integers ⁇ 0, and 0 or at least 1 methylene unit of W is independently replaced by -Cyr-, -N(R wx )C(O)-, - C(O)N(R wx )-, -C(O)-, -OC(O)-, -C(O)O-,
  • the linker-payload compound is of the formula: .
  • the linker-payload compound may be of the formula 5 P129624PCT wherein, LG is a leaving group; 5 each Z is independently selected from a direct bond, a carbon-carbon triple bond, a carbon- carbon double bond, and amido (preferably selected from a direct bond, a carbon-carbon triple bond, and a carbon-carbon double bond); 10 Rx and Ry are each independently selected from H and C1-4 alkyl; each m is independently selected from 0, 1, 2, 3, 4, 5 and 6; y1 is selected from any integer between 1 and 6 (such as 4, 5, and 6); each y2 is independently selected from any integer between 0 and 15 (such as 6-15); each y3 is independently selected from 1, 2, and 3; 15 each y4 is independently selected from 0 and 1; position 1 is attached to LG, and position 2 is attached to L 2 or L 3 ; L 2 is absent or present, and when L 2 is present, L 2 is selected from:
  • R 15 r, r 1 , r 1a and r 1b are each independently 0, 1, 2, 3, 4 or 5;
  • R m1 , R n1 , R m1a , R n1a , R m1b and R n1b are each independently H, C 1-6 alkyl or -COOR x1 , wherein, R x1 is C 1-6 alkyl; or, R m1 and R n1 , R m1a and R n1a , and R m1b and R n1b , together with the nitrogen atom to which they are both attached, form a 5-6 membered heterocyclic ring, and of said 5-6 membered 20 heterocyclic ring, the heteroatom is selected from 1 or 2 N atoms; the said 5-6 membered heterocyclic ring is optionally substituted with one or more R 0’ ;
  • P129624PCT R z is selected from C 1-6 alkyl;
  • LG is selected from halogen, sulfone group, a tertiary amine salt group diazonium salt group, -OMs, MeSO 2 -, and CF3SO3-; In some embodiments, LG is selected from F, Cl and MeSO2- and the tertiary amine salt group is selected from Me3N+ and Et3N+; In some embodiments, LG is selected from F and MeSO2-. 10 In some embodiments, the linker-payload compound is of the formula: Nucleic acid sequence(s) The present invention provides one or more nucleic acid sequence(s) encoding the binding molecule as defined herein.
  • the present invention provides one or more nucleic 15 acid sequence(s) capable of expressing the binding molecule as defined herein.
  • the present invention provides one or more nucleic acid sequence(s) encoding (i.e. capable of expressing) the antibody according to the invention.
  • the terms “polynucleotide”, “nucleotide”, and “nucleic acid” are intended to be synonymous with each other.
  • the nucleic acid sequence(s) may be RNA or DNA sequences, 20 or a mixture of RNA and DNA sequences. P129624PCT In an embodiment, the nucleic acid sequence(s) are one or more DNA sequences, such as cDNA sequences.
  • the nucleic acid sequence is a DNA sequence, such as a cDNA sequence.
  • the nucleic acid sequence(s) are RNA sequences, such as mRNA sequences.
  • the nucleic acid sequence is an RNA sequence, 5 such as an mRNA sequence.
  • the nucleic acid sequence(s) may be single-stranded or may be double-strandedp.
  • the nucleic acid sequence(s) may be, for example, genomic, recombinant, mRNA or cDNA.
  • the nucleic acid sequence(s) may comprise synthetic nucleotides and/or modified nucleotides. These synthetic nucleotides and/or modified nucleotides may enhance in vivo activity and/or 10 stability.
  • nucleic acid sequences Due to the redundancy of the genetic code, variations in nucleic acid sequences are possible that encode for the same polypeptide. These variations in nucleic acid sequences are encompassed by the present invention. Therefore, multiple nucleic acid sequence(s) are envisaged, each of which may be different, but which still encode a binding molecule or 15 antibody according to the present invention. It is known in the art how to design and produce such nucleic acid sequences.
  • the nucleic acid sequence(s) may be codon optimised for production in the host cell of choice.
  • nucleic acid sequence(s) may be operably linked to further sequence(s) such as control sequence(s), e.g.
  • the nucleic acid sequence(s) may be in the form of one or more expression cassettes.
  • the nucleic acid sequences may be suitable for expression in prokaryotic cells or in eukaryotic cells, such as mammalian cells. Any promoter may be used, such as a strong promoter that is functional in prokaryotic cells or 25 in eukaryotic cells. Suitable promoters will be known in the art.
  • the promoter may be a constitutive promoter.
  • the promoter may be a tissue specific promoter.
  • Vector The present invention provides a vector comprising the one or more nucleic acid sequence(s) of the invention.
  • the vector may comprise a polynucleotide comprising a nucleic acid sequence or sequences encoding the binding molecule as defined herein.
  • the vector may comprise a P129624PCT polynucleotide comprising a nucleic acid sequence or sequences encoding the antibody according to the invention.
  • the vector may be used to introduce nucleic acid sequence(s) according to the invention into a cell so that the cell expresses and/or produces the binding molecule as defined herein or 5 the antibody according to the invention.
  • the term “vector” may be considered interchangeable with the term “expression vector” and “expression construct”.
  • the vector may be any vector that is suitable for introducing and/or expressing a nucleic acid sequence in a cell.
  • the vector may comprise regulatory sequences, enhancer sequences and/or promoter sequences that promote 10 expression of a nucleic acid sequence in a cell.
  • the vector according to the invention may be any agent capable of delivering nucleic acid sequence(s) according to the invention to a cell and/or expressing nucleic acid sequence(s) according to the invention in a cell. Examples of suitable vectors include but are not limited to plasmids, cosmids, phages, viruses or artificial chromosomes. 15
  • the vector may be a plasmid or a viral vector.
  • the vector may be a retroviral vector or a lentiviral vector. The vector may be capable of transfecting or transducing a cell.
  • the present invention provides a cell comprising one or more nucleic acid sequence(s) 20 according to the invention, or a vector according to the invention.
  • the present invention provides a cell comprising a binding molecule as defined herein.
  • the present invention provides a cell comprising a BPC according to the invention.
  • the present invention provides a cell comprising an antibody according to the invention.
  • the polynucleotide or vector may, for example, be introduced into a cell by transduction or 25 transfection in vitro or ex vivo.
  • the present invention also provides a method for making a cell according to the invention comprising the step of introducing the one or more nucleic acid(s) according to the P129624PCT invention, or the vector according to the invention into said cell.
  • the nucleic acid(s) may be introduced as described herein.
  • the cell may be capable of expressing the binding molecule as defined herein. 5
  • the cell may be capable of expressing the antibody according to the invention.
  • the cell may be capable of producing the binding molecule as defined herein or antibody according to the invention.
  • the cell may be capable of expressing and/or producing the binding 10 molecule or antibody as defined herein when the cell is cultured under suitable conditions.
  • the present invention also provides a method for producing the binding molecule as defined herein, wherein the method comprises the steps of: (i) introducing one or more nucleic acid sequence(s) according to the invention, or a vector according to the invention into a cell; and 15 (ii) expressing the binding molecule thereof in the cell.
  • the present invention also provides a method for producing the antibody according to the invention, wherein the method comprises the steps of: (i) introducing one or more nucleic acid sequence(s) according to the invention, or a vector according to the invention into a cell; and 20 (ii) expressing the binding molecule thereof in the cell.
  • the one or more nucleic acid sequence(s) or vector may be introduced into the cell by transduction or transfection in vitro or ex vivo.
  • culturing the cell under 25 suitable conditions may result in the cell expressing and/or producing the binding molecule or antibody as defined herein.
  • the method for producing the binding molecule or antibody as defined herein thereof may further comprise step (iii) harvesting the binding molecule or antibody or P129624PCT fragment thereof from the cell or cell culture supernatant of the cell. It will be understood that the binding molecule or antibody may be harvested from the cell.
  • the binding molecule or antibody may be harvested from supernatant of the cell, for example when the binding molecule or antibody is released out of 5 the cell into the cell culture medium that the cell is cultured in.
  • the cell may be a prokaryotic cell or a eukaryotic cell.
  • the cell may be a bacterial cell, a fungal cell, a yeast cell, a plant cell or an animal cell.
  • the cell may be a mammalian cell or an insect cell.
  • the cell may be a human cell.
  • Composition The present invention also provides a composition which comprises one or more BPCs according to the invention.
  • the present invention also provides a pharmaceutical composition comprising the binding molecule as defined herein. 15
  • the drug-antibody ratio (DAR) of the BPCs according to the invention may vary. Consequently, the composition may comprise a mixture of BPCs having a number of different DARs, and may therefore have an average DAR which is non-integral.
  • average DAR and “average connection number” are synonymous.
  • the average DAR is an integer or decimal from about 1 to about 16.
  • the average DAR is an integer or decimal of about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15 or about 16.
  • the average DAR is an integer or decimal from about 4 to about 12. In some embodiments, the average DAR is an integer or decimal from about 6 to about 10. In some embodiments, the average DAR is an integer or 25 decimal from about 7 to about 9. In some embodiments, the average DAR is an integer or decimal from about 7.5 to about 8.5. In some embodiments, the average DAR is an integer or decimal from about 7.8 to about 8.2. In some embodiments, the average DAR is about 6.0. In some embodiments, the average DAR is about 6.1. In some embodiments, the average DAR is about 6.2. In some P129624PCT embodiments, the average DAR is about 6.3. In some embodiments, the average DAR is about 6.4.
  • the average DAR is about 6.5. In some embodiments, the average DAR is about 6.6. In some embodiments, the average DAR is about 6.7. In some embodiments, the average DAR is about 6.8. In some embodiments, the average DAR is 5 about 6.9. In some embodiments, the average DAR is about 7.0. In some embodiments, the average DAR is about 7.1. In some embodiments, the average DAR is about 7.2. In some embodiments, the average DAR is about 7.3. In some embodiments, the average DAR is about 7.4. In some embodiments, the average DAR is about 7.5. In some embodiments, the average DAR is about 7.6. In some embodiments, the average DAR is about 7.7.
  • the average DAR is about 7.8. In some embodiments, the average DAR is about 7.9. In some embodiments, the average DAR is about 8.0. In some embodiments, the average DAR is about 8.1. In some embodiments, the average DAR is about 8.2. In some embodiments, the average DAR is about 8.3. In some embodiments, the average DAR is about 8.4. In some embodiments, the average DAR is about 8.5. In some embodiments, the 15 average DAR is about 8.6. In some embodiments, the average DAR is about 8.7. In some embodiments, the average DAR is about 8.8. In some embodiments, the average DAR is about 8.9. In some embodiments, the average DAR is about 9.0. In some embodiments, the average DAR is about 9.1.
  • the average DAR is about 9.2. In some embodiments, the average DAR is about 9.3. In some embodiments, the average DAR is 20 about 9.4. In some embodiments, the average DAR is about 9.5. In some embodiments, the average DAR is about 9.6. In some embodiments, the average DAR is about 9.7. In some embodiments, the average DAR is about 9.8. In some embodiments, the average DAR is about 9.9. In some embodiments, the DAR is about 10.0. In some embodiments, the composition comprises one or more conjugates having the 25 structure shown as formula (III-D’):
  • the composition comprises one or more conjugates having the structure shown as Formula (IVA’): or a pharmaceutically acceptable salt thereof, wherein, 10 q’ represents an average connection number, and is an integer or decimal from 1 to 16, and BM is a binding molecule as defined herein.
  • the composition comprises one or more conjugates having the structure shown as formula (II-Dx’): , wherein: 5 BM, L, L2, ring A, X and L1 are as defined above for formula (III-D), either in its broadest aspect or a preferred aspect; and q’ represents an average connection number, and is an integer or decimal from 1 to 16.
  • the composition comprises one or more conjugates having the structure shown as formula (IVB’) 10 or a pharmaceutically acceptable salt thereof, wherein, q’ represents an average connection number, and is an integer or decimal from 1 to 16; and BM is a binding molecule as defined herein.
  • the composition comprises one or more conjugates having the structure shown as formula I’: wherein: 5 BM, L1, L2, L3, L4, W, X, R1, R2 and R3 are as defined above for formula (I’), either in its broadest aspect or a preferred aspect; and q’ is an average connection number and is an integer or decimal from 1 to 16.
  • the composition comprises one or more conjugates having the structure shown as formula (IVC‘): 10 or a pharmaceutically acceptable salt thereof, wherein, q’ represents an average connection number, and is an integer or decimal from 1 to 16; and BM is a binding molecule as defined herein.
  • the average P129624PCT connection number q’ is an integer or decimal from about 1 to about 16. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is an integer or decimal of about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15 or about 16.
  • the average connection number q’ is an integer or decimal from about 4 to about 12. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is an integer or decimal from about 6 to about 10. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is an integer or decimal from about 7 10 to about 9.
  • the average connection number q’ is from about 7.5 to about 8.5. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is from about 7.8 to about 8.2. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average 15 connection number q’ is about 6.0.
  • the average connection number q’ is about 6.1. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 6.2. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 6.3.
  • the average connection number q’ is about 6.4. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 6.5. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 6.6.
  • the average connection number q’ is about 6.7. In some embodiments of 25 formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 6.8. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 6.9.
  • the average connection number q’ is about 7.0. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 30 7.1. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 7.2.
  • the average connection number q’ is about 7.3. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 7.4. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average P129624PCT connection number q’ is about 7.5.
  • the average connection number q’ is about 7.6. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 7.7. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average 5 connection number q’ is about 7.8.
  • the average connection number q’ is about 7.9. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 8.0. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 8.1.
  • the average connection number q’ is about 8.2. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 8.3. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 8.4.
  • the average connection number q’ is about 8.5. In some embodiments of 15 formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 8.6. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 8.7.
  • the average connection number q’ is about 8.8. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 20 8.9. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 9.0.
  • the average connection number q’ is about 9.1. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 9.2. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average 25 connection number q’ is about 9.3.
  • the average connection number q’ is about 9.4. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 9.5. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 9.6.
  • the average connection number q’ is about 9.7. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 9.8. In some embodiments of formulae (III-D’), (IVA’), (II-Dx’), (IVB’), (I’) or (IVC’), the average connection number q’ is about 9.9.
  • the average connection number q’ is about 10.0.
  • P129624PCT Pharmaceutical Composition
  • the present invention also provides a pharmaceutical composition which comprises the BPC according to the invention, together with a pharmaceutically acceptable carrier, diluent or excipient.
  • the present invention also provides a pharmaceutical composition which comprises the antibody according to the invention, together with a pharmaceutically acceptable carrier, diluent or excipient.
  • the present invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising the binding molecule as defined herein, the one or more nucleic acid(s) according to the invention, or the 10 vector according to the invention, together with a pharmaceutically acceptable carrier, diluent or excipient.
  • the compositions described herein may further comprise one or more selected from this list consisting of: an adjuvant, salt, active polypeptide, compound, component and active agent.
  • Compositions typically should be sterile and stable under the conditions of manufacture and storage.
  • the composition according to the invention may be produced using current good manufacturing practices (CGMP).
  • the pharmaceutical composition may be formulated to be suitable for administration to a patient in order to prevent and/or treat disease.
  • compositions can be 20 formulated for administration by different routes, for example, for oral, parenteral, topical, inhalative, intravenous, intramuscular, rectal, sublingual, transdermal, subcutaneous, intratumoral application routes, according to their chemical and physical properties.
  • the pharmaceutical composition may be in the form of a tablet, a coated tablet, powder, granulate, a pellet, a capsule, an effervescent tablet or a transdermal therapeutic system.
  • the 25 pharmaceutical composition may be in the form of a liquid composition, selected from the group consisting of a solution, a syrup, an infusion, an extract, a solution for intravenous application, or a solution for infusion.
  • the pharmaceutical composition may be in the form of a semisolid composition such as an emulsion, a suspension, a cream, a lotion, a gel, a globule, a buccal tablet or a suppository.
  • carrier may refer to a diluent, adjuvant, excipient, or vehicle.
  • Such carriers can be sterile liquids, such as saline solutions in water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil.
  • a sterile saline solution is a preferred carrier. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid 5 carriers.
  • Suitable excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like.
  • the composition if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents.
  • the composition of the invention can be formulated as neutral or salt 10 forms.
  • Salts include those formed with free amino groups such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, etc., and those formed with free carboxyl groups such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, etc.
  • the salt may comprise a metal cation, such as a sodium salt or a 15 potassium salt.
  • the composition may comprise an aqueous diluent or solvent.
  • the aqueous diluent or solvent may be a phosphate buffered saline solution, such as a sterile phosphate buffered saline solution.
  • the composition may comprise one or more vesicles, nanoparticles, 20 lipid nanoparticle (LNPs), liposomes or polymeric mixtures.
  • the composition may enable delivery of a nucleic acid(s) according to the invention and/or a vector according to the invention to a cell.
  • Kit The present invention provides a kit comprising the BPC according to the invention. 25
  • the present invention provides a kit comprising the antibody according to the invention.
  • the present invention provides a kit comprising the composition according to the invention.
  • the kit may optionally comprise instructions for using the kit to target the one or more payload(s) to a cell expressing CA19-9.
  • P129624PCT Methods and uses The present invention provides an in vitro method comprising contacting a cell with the BPC according to the invention.
  • the present invention provides an in vitro method comprising contacting a cell with the 5 antibody according to the invention.
  • the invention provides a method of treating or diagnosing a disease, comprising administering the BPC according to the invention to a subject.
  • the invention provides a method of treating or diagnosing a disease, comprising administering the antibody according to the invention to a subject.
  • 10 The invention provides a method of treating or diagnosing a disease, comprising administering the composition according to the invention to a subject.
  • the invention provides for use of the BPC according to the invention or the antibody according to the invention or the composition according to the invention for the manufacture of a medicament.
  • the invention provides the BPC according to the invention or the antibody according to the invention or the composition according to the invention for use as a medicament.
  • the invention provides the BPC according to the invention for use in a method of therapy or a diagnostic method.
  • the invention provides the antibody according to the invention for use in a method of therapy 20 or a diagnostic method.
  • the invention provides the composition according to the invention for use in a method of therapy or a diagnostic method.
  • the disease is cancer.
  • the medicament may be for use in the treatment of cancer.
  • the method may be a method of treating, preventing or diagnosing cancer.
  • the cancer may express CA19-9.
  • the expression of CA19-9 is increased compared to the expression of CA19-9 by the same non-cancerous tissue or cells.
  • the cancer is gastrointestinal cancer, pancreatic cancer, ovarian P129624PCT cancer, colorectal cancer, stomach cancer, oesophageal cancer, endometrial cancer, breast cancer, bile duct carcinoma, transitional cell carcinoma, or hepatocellular carcinoma.
  • the pancreatic cancer is pancreatic ductal adenocarcinoma (PDAC).
  • the term “CA19-9-positive cancer” means a cancer involving 5 cancer cells expressing CA19-9, preferably on the surface of said cancer cells.
  • Cell surface is used in accordance with its normal meaning in the art, and thus includes the outside of the cell which is accessible to binding by proteins and other molecules. For example, a transmembrane protein having one or more extracellular portions is considered as being expressed on the cell surface. 10 CA19-9 is expressed on the surface of cells if it is located at the surface of said cells and is accessible to binding by CA19-9-specific antibodies added to the cells which have not been disrupted. According to the invention, the term “disease” refers to any pathological state, including cancer, in particular those forms of cancer described herein. Any reference herein to cancer 15 or particular forms of cancer also includes cancer metastasis thereof. In a preferred embodiment, a disease to be treated according to the present application involves cells expressing CA19-9.
  • “Diseases associated with cells expressing CA19-9” or similar expressions means according to the invention that CA19-9 is expressed in cells of a diseased tissue or organ. 20
  • expression of CA19-9 in cells of a diseased tissue or organ is increased compared to the state in a healthy tissue or organ.
  • An increase refers to an increase by at least 10%, in particular at least 20%, at least 50%, at least 100%, at least 200%, at least 500%, at least 1000%, at least 10000% or even more.
  • expression is only found in a diseased tissue, while expression in a 25 corresponding healthy tissue is repressed.
  • CA19-9 is expressed in pancreatic cancer tissue while expression is not detectable in non-cancerous pancreatic tissue.
  • diseases associated with cells expressing CA19-9 include cancer diseases.
  • cancer diseases preferably are those wherein the cancer cells express CA19-9.
  • P129624PCT As used herein, a “cancer disease” or “cancer” includes a disease characterized by aberrantly regulated cellular growth, proliferation, differentiation, adhesion, and/or migration.
  • cancer cell is meant an abnormal cell that grows by a rapid, uncontrolled cellular proliferation and continues to grow after the stimuli that initiated the new growth cease.
  • a “cancer 5 disease” is characterized by cells expressing CA19-9 and a cancer cell expresses CA19-9.
  • a cell expressing CA19-9 preferably is a cancer cell, preferably of the cancers described herein.
  • a “carcinoma” is a malignant tumor derived from epithelial cells.
  • “Adenocarcinoma” is a cancer that originates in glandular tissue. This tissue is also part of a larger tissue category known as epithelial tissue. Epithelial tissue includes skin, glands and a 10 variety of other tissue that lines the cavities and organs of the body. Epithelium is derived embryologically from ectoderm, endoderm and mesoderm. To be classified as adenocarcinoma, the cells do not necessarily need to be part of a gland, as long as they have secretory properties. This form of carcinoma can occur in some higher mammals, including humans.
  • adenocarcinomas tend to resemble the glandular tissue that they 15 are derived from, while poorly differentiated may not.
  • a pathologist By staining the cells from a biopsy, a pathologist will determine whether the tumor is an adenocarcinoma or some other type of cancer.
  • Adenocarcinomas can arise in many tissues of the body due to the ubiquitous nature of glands within the body. While each gland may not be secreting the same substance, as long as there is an exocrine function to the cell, it is considered glandular and its malignant form is 20 therefore named adenocarcinoma. Malignant adenocarcinomas invade other tissues and often metastasize given enough time to do so.
  • the pancreas an organ of endodermal derivation, is the key regulator of protein and carbohydrate digestion and glucose homeostasis.
  • the exocrine pancreas (80% of the tissue mass of the organ) is composed of a branching network of acinar and duct cells that produce 25 and deliver digestive enzymes into the gastrointestinal tract.
  • the acinar cells which are organized in functional units along the duct network, synthesize and secrete enzymes into the ductal lumen in response to cues from the stomach and duodenum. Within the acinar units near the ducts are centroacinar cells.
  • pancreas which regulates metabolism and glucose homeostasis through the secretion of hormones into the bloodstream, is 30 composed of four specialized endocrine cell types gathered together into clusters called Islets of Langerhans.
  • Pancreatic cancer is a malignant neoplasm originating from transformed cells arising in tissues P129624PCT forming the pancreas.
  • Pancreatic cancer includes adenocarcinomas (tumors exhibiting glandular architecture) arising within the exocrine component of the pancreas and neuroendocrine carcinomas arising from islet cells.
  • pancreatic cancer pancreatic cancer
  • ductal adenocarcinoma pancreatic ductal adenocarcinoma
  • PDAC pancreatic ductal adenocarcinoma
  • PDAC primarily exhibits a glandular pattern with duct-like structures and varying degrees of cellular atypia and differentiation.
  • Less 10 common subtypes of PDAC include colloid, adenosquamous, or sarcomatoid histology.
  • pancreas Often within an individual tumor, there are regional differences in histology, tumor grade, and degree of differentiation. Even the smallest primary lesions commonly exhibit perineural and lympho- vascular invasion, suggesting a propensity for early distant spread.
  • the second most common type of exocrine pancreas cancer is mucinous. Mucinous 15 adenocarcinoma produces a large volume of mucin that results in a cystic appearance on imaging studies.
  • Pancreatic neuroendocrine tumors form in hormone-making cells (islet cells) of the pancreas. Acinic cell neoplasms arise from the acinar cells of the pancreas.
  • cancer also includes cancer metastasis of a primary 20 tumor such as primary pancreatic cancer.
  • a primary 20 tumor such as primary pancreatic cancer.
  • this also includes metastasis of the pancreatic cancer, for example metastasis to the lung, liver and/or lymph nodes.
  • metastasis is meant the spread of cancer cells from its original site to another part of the body. The formation of metastasis is a very complex process and depends on detachment of 25 malignant cells from the primary tumor, invasion of the extracellular matrix, penetration of the endothelial basement membranes to enter the body cavity and vessels, and then, after being transported by the blood, infiltration of target organs.
  • the term “metastasis” according to the invention relates to “distant metastasis” which relates to a metastasis which is remote from the primary tumor and the regional lymph node system.
  • the term “metastasis” according to the P129624PCT invention relates to lymph node metastasis.
  • metastasis One particular form of metastasis which is treatable using the therapy of the invention is metastasis originating from pancreatic cancer as primary site.
  • pancreatic cancer metastasis is metastasis into lymph nodes, metastasis into lung and/or metastasis into liver.
  • a refractory cancer is a malignancy for which a particular treatment is ineffective, which is either initially unresponsive to treatment, or which becomes unresponsive over time.
  • treat is meant to administer a compound or composition or a combination of compounds or compositions to a subject in order to prevent or eliminate a disease, including reducing the size of a tumor or the number of tumors in a subject; arrest or slow a disease in a subject; 10 inhibit or slow the development of a new disease in a subject; decrease the frequency or severity of symptoms and/or recurrences in a subject who currently has or who previously has had a disease; and/or prolong, i.e., increase the lifespan of the subject.
  • treatment of a disease includes curing, shortening the duration, ameliorating, preventing, slowing down or inhibiting progression or worsening, or preventing 15 or delaying the onset of a disease or the symptoms thereof.
  • patient means according to the invention a subject for treatment, in particular a diseased subject, including human beings, nonhuman primates or other animals, in particular mammals such as cows, horses, pigs, sheep, goats, dogs, cats or rodents such as mice and rats. In a particularly preferred embodiment, a patient is a human being.
  • ADCC and CDC assays with 5B1-Fc-inert Mab and ADC variants For ADCC (antibody-dependent cellular cytotoxicity) assay, COLO 205 cell line served as target positive cell line. Human PBMCs from a healthy donor were used as effector cells (E:T ratio 40:1). Target cells and effector cells were incubated for 12 hours with 5B1 mAb and 5B1- 25 ADCs which have the formula (IVC) as defined above wherein BM is one of a number of antibodies with different Fc-silencing mutations (LALA, LALA-KA, LALA-deltaA & STR) in different concentrations (25000 ng/mL – 0.01 ng/mL).
  • IVC formula
  • the specific cell lysis was determined using a luciferase-based assay.
  • CDC complement-dependent cytotoxicity
  • CA-19-9+ CHO-K1 transfectants (solid 30 line) served as target cells.
  • Target cells were incubated with human serum (25 % final P129624PCT concentration) and 5B1 mAb and 5B1-ADCs with different Fc silencing mutations (LALA, LALA-KA, LALA-deltaA & STR) at different concentrations (250000 ng/mL – 15.7 ng/mL) for 4 hours.
  • CDC was determined using a luciferase-based assay.
  • ADC-1 has the formula (IVC) as defined above, wherein BM is 5B1 antibody, and q’ has an average value of 8.0. 10
  • a CA19.9-positive human pancreatic ductal adenocarcinoma (BxPC3) xenograft mouse model was utilized.
  • a treatment regimen comprising 15 seven intraperitoneal (i.p.) bolus injections once weekly was initiated one day later applying ADC-1 in flat doses of 3 mg/kg or 8 mg/kg (Figure 2A).
  • a target irrelevant ctrl A target irrelevant ctrl.
  • IgG-ADC (Comparative ADC-1) , which has the formula (IVC) wherein BM is a target irrelevant IgG and the 5B1 monoclonal antibody lacking any payload-linker moieties (AB-1) served as controls with a dose of 8 mg/kg respectively.
  • 5B1-ADC treatment lead to potent elimination of advanced human PDAC tumor xenograft in mice ( Figure 2B).
  • P129624PCT 5B1-ADC treatment is well tolerated in mice and leads to survival benefits. Body weights of mice were examined twice per week using a laboratory scale. No weight loss occurred in any treatment or control group (Figure 2C).
  • ADC-2 has the formula (IVA) as defined above, wherein BM is 5B1 antibody, and q’ has an 10 average value of 7.99.
  • the linker-payload portion of ADC-2 is referred to in this Example as “Linker 2”).
  • ADC-3 has the formula (IVB) as defined above, wherein BM is 5B1 antibody, and q’ has an average value of 8.00.
  • Linker 1 The linker-payload portion of ADC-3 is referred to in this Example as “Linker 1”). 15
  • BxPC3 human pancreatic ductal adenocarcinoma
  • a treatment regimen comprising 20 seven intraperitoneal (i.p.) bolus injections once weekly was initiated two days later applying ADC-2 or ADC-3 in flat doses of 3 mg/kg and 8 mg/kg respectively (Figure 3A).
  • a target irrelevant IgG-ADC which has the formula (IVA) wherein BM is a target irrelevant IgG and the payload-linker portion is Linker 2 (Comparative ADC-2) and the 5B1 monoclonal antibody lacking any payload-linker moieties (AB-1) were used as controls with a dose of 8 mg/kg 25 respectively.
  • P129624PCT Tumour growth was monitored three times a week. 5B1-ADC treatment lead to potent elimination of advanced human PDAC tumor xenograft in mice.
  • SK-OV-3 cells which do not express the target, served as negative control.
  • 5B1-ADC Fc was added to the cells at concentrations ranging from 0.046 to 300 nM.
  • the unconjugated 5B1-mAb Fc-inert served as positive control, while a non- 15 target specific hIgG1 and an isotype ADC (ADC-IC) were utilized as negative controls.
  • ADC-IC isotype ADC
  • the samples were incubated for 60 minutes.
  • a PE-conjugated anti-human IgG Fc antibody was used as a secondary antibody.
  • 5B1-ADC Fc-inert showed specific and concentration- dependent binding to the target-positive COLO 205, HT-1197 and BxPC3 cells (Fig.4A-C).
  • 5B1-ADC Fc-inert was serially diluted (63 to 25 250,000 ng/mL) and added the target cells.5B1-mAb Fc-comp was used as a positive control. Pooled normal human serum was then added to a final concentration of 25% and the samples were incubated for 4 h at 37°C and 5% CO 2 . Luciferase activity was measured as readout for cell lysis. The CDC activity of 5B1-ADC Fc-inert was reduced compared to 5B1-mAb Fc-comp P129624PCT (Fig.5A).
  • luciferase-expressing COLO 205 cells (COLO 205_luc) served as target- positive cells.5B1-ADC Fc-inert was serially diluted (0.01 to 25,000 ng/mL) and added to the target cells.5B1-mAb Fc-comp was used as a positive control.
  • Human PBMCs from a healthy 5 donor served as effector cells and were added at an E:T ratio of 40:1. The samples were incubated for 12 h at 37°C with 5% CO 2 . Luciferase activity was measured as readout for cell lysis. 5B1-ADC Fc-inert showed no ADCC activity, even at high concentrations.
  • 5B1-mAb Fc-comp mediated potent, dose-dependent lysis of the target cells (Fig.5B).
  • ADCP activity of Fc-inert 5B1-ADC and 5B1-mAb and Fc-competent 5B1-mAb in 10 human cancer cell lines The ADCP activity of 5B1-ADC Fc-inert was assessed using flow cytometry across four human cancer cell lines, including CA19-9 high-expressing COLO 205 cells, medium-expressing BxPC3 and HT-1197 cells. SK-OV-3 cells, which do not express the target, served as negative control.
  • M2c macrophages derived from human PBMCs served as effector cells and were15 added to the CellTrace Far Red dye labeled target cells at an E:T ratio of 4:1.5B1-ADC Fc- inert was added at concentrations ranging from 0.001 to 101 nM and incubated for four hours.
  • the unconjugated 5B1-mAb Fc-inert served as negative control
  • 5B1- ADC Fc-comp served as positive control.
  • An anti-human CD14 antibody was used to detect the effector cells via flow cytometry.
  • Anti-proliferation effect of Fc-inert 5B1-ADC and control items in human cancer cell lines The anti-proliferation activity of 5B1-ADC Fc-inert was assessed in CA19-9 high-expressing COLO 205 cells, medium-expressing BxPC3 and HT-1197 cells. SK-OV-3 cells, which do not 30 express the target, served as negative control.
  • the unconjugated 5B1-mAb Fc-inert, ADC-IC and the free toxin payload (YL0010014) served as controls.
  • Target cells were incubated with the test and control items at concentrations ranging from 0.001 nM to 200 nM for five days.
  • P129624PCT Cell viability was measured using the CellTiter-Glo luminescent cell viability assay.5B1-ADC Fc-inert exhibited potent anti-proliferative effects in target-expressing COLO 205 and BxPC3 cell lines (Fig.7A, C). In contrast, 5B1-ADC Fc-inert did not show any anti-proliferative effects on the target-negative SK-OV-3 cell line, indicating the target-specific cytotoxicity of 5B1-ADC 5 Fc-inert. For the HT-1197 cell line, no inhibition curve could be generated for either 5B1-ADC Fc-inert or YL0010014, indicating poor susceptibility of this cell line to the payload.
  • the ADC-IC exhibited non-specific anti-proliferative effects on COLO 205, BxPC3, and SK-OV-3 cells (Fig.7D).
  • Internalization of Fc-inert 5B1-ADC and control items by human cancer cell lines 10 The internalization of 5B1-ADC Fc-inert was assessed in CA19-9 high-expressing COLO 205 cells and medium-expressing BxPC3 and HT-1197 cells by flow cytometry. SK-OV-3 cells, which do not express the target, served as negative control.
  • the unconjugated 5B1-mAb Fc- inert, ADC-IC and a human IgG1 isotype-mAb served as controls.
  • Test and control items were labeled with the pH-sensitive dye; pH-rodo.
  • Cell lines were incubated with 40 nM of labeled 15 test and control items and internalization was measured at indicated time points via flow cytometry.
  • the internalization of 5B1-ADC Fc-inert was detectable after three hours of incubation and further increased reaching a maximal internalization rate of nearly 100% at 12 hours (Fig 8A).
  • the internalization of 5B1-ADC Fc-inert was also detectable after three hours of incubation and further increased over the 24 20 hour time course reaching a moderate internalization of 45-60% (Fig 8B, C).
  • the 5B1-mAb Fc-inert showed similar internalization rates in all cell lines indicating that the addition of the linker-payload has no effect on the internalization.
  • Low levels of non-specific internalization were observed in COLO 205 and BxPC3 cells treated with ADC-IC and a human IgG1 isotype- mAb at 12 and 24 hours.
  • No internalization was observed in target-negative SK-OV-3 cells 25 indicating target-specific internalization of 5B1-ADC Fc-inert and 5B1-mAb Fc-inert (Fig 8D).
  • SK-OV-3-luc cell monocultures served as negative control.
  • Cell cultures were incubated with 5B1-ADC Fc-inert and ADC-IC at concentrations ranging from 0.0001 nM to 40 nM for five days.
  • Bystander activity was measured by luciferase activity.5B1-ADC Fc-inert exhibited potent bystander killing activity 5 effects in target-negative SK-OV-3-luc cells co-cultured with target-expressing COLO 205 (Fig. 11A) and BxPC3 (Fig.11B) cell lines with corresponding ReIC50 values of 1.29 nM and 0.05 nM, respectively.
  • 5B1-ADC Fc-inert did not show any anti-proliferative effects on the target-negative SK-OV-3 monoculture, indicating the target-specific bystander killing activity of 5B1-ADC Fc-inert in co-cultures (Fig.11C).
  • the ADC-IC exhibited non-specific bystander killing activity in SK-OV-3-luc cells co- cultured with BxPC3 cells (Fig.11B) and SK-OV-3-luc monocultures (Fig.11C).
  • 5B1-ADC Fc-inert treatment is well tolerated, leads to potent elimination of advanced xenograft tumors, and provides survival benefits in mice (BxPC3)
  • BxPC3 a CA19-9-positive human 15 pancreatic ductal adenocarcinoma
  • a treatment regimen comprising six intravenous (i.v.) bolus injections once weekly was initiated three days later applying 5B1-ADC 20 Fc-inert in flat doses of 3 mg/kg or 8 mg/kg.
  • the unconjugated Fc-competent 5B1 monoclonal antibody (5B1-mAb Fc-comp) and the payload-conjugated Fc-competent 5B1 ADC (5B1-ADC Fc-comp) were serving as controls with a dose of 8 mg/kg, while a third control group received DPBS. Body weights of mice were examined twice per week using a laboratory scale.
  • mice were stratified into treatment and control groups.
  • a treatment regimen comprising six intravenous (i.v.) bolus injections once weekly was initiated one day later applying 5B1-ADC Fc-inert in flat doses of 1, 3 and 8 mg/kg respectively.
  • the control group received DPBS. No 10 weight loss occurred in any treatment or control group. No other clinical signs occurred during 5B1-ADC Fc-inert treatment confirming a good tolerability of 5B1-ADC Fc-inert in mice (Fig. 13A).
  • 5B1-ADC Fc-inert treatment at a dose of 8 mg/kg resulted in a significant anti-tumor effect compared to the DPBS control group.
  • 5B1-ADC Fc-inert treatment at doses of 3 mg/kg showed a delay in tumor growth (Fig.13B).
  • 5B1-ADC Fc-inert treatment at 8 mg/kg conferred 15 significant survival benefits over the DPBS control group (Fig.13C).
  • 5B1-ADC Fc-inert treatment is well tolerated and leads to potent elimination of advanced xenograft tumors (CAPAN-2)
  • CAPAN-2 advanced xenograft tumors
  • CAPAN-2 CA19-9-positive human pancreatic ductal adenocarcinoma
  • a treatment regimen comprising four intravenous (i.v.) bolus injections once weekly was initiated three days later applying 5B1-ADC Fc-inert in flat doses of 0.3, 1, 3 or 8 mg/kg, respectively.
  • the control group received DPBS.
  • Body weights of mice 25 were examined twice per week using a laboratory scale. Weight loss occurred in all groups at the start of treatment. However, the body weight recovered at later time points. No clinical signs were observed in any of the treatment groups confirming a good tolerability of 5B1-ADC Fc-inert in mice (Fig.14A).
  • Treatment with 1, 3 and 8 mg/kg 5B1-ADC Fc-inert resulted in a significant reduction in tumor volumes compared to the DPBS control groups (Fig. 14B).
  • a binding molecule-payload conjugate comprising a binding molecule and one or more payload moieties; wherein: a) the binding molecule is an antibody or a fragment thereof and comprises heavy chain 5 complementarity determining regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3 according to the IMGT numbering scheme, wherein: HCDR1 comprises an amino acid sequence according to SEQ ID NO: 1, HCDR2 comprises an amino acid sequence according to SEQ ID NO: 2, HCDR3 comprises an amino acid sequence according to SEQ ID NO: 3, LCDR1 comprises an amino acid 10 sequence according to SEQ ID NO: 4, LCDR2 comprises an amino acid sequence according to SEQ ID NO: 5, LCDR3 comprises an amino acid sequence according to SEQ ID NO: 6, optionally wherein one or more of the CDRs comprises one
  • a binding molecule-payload conjugate comprising a binding molecule and one or more payload moieties; wherein: a) the binding molecule is an antibody or a fragment thereof and comprises heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity 20 determining regions (LCDRs) 1-3 according to the Kabat numbering scheme, wherein: HCDR1 comprises an amino acid sequence according to SEQ ID NO: 15, HCDR2 comprises an amino acid sequence according to SEQ ID NO: 16, HCDR3 comprises an amino acid sequence according to SEQ ID NO: 17, LCDR1 comprises an amino acid sequence according to SEQ ID NO: 18, LCDR2 comprises an amino acid 25 sequence according to SEQ ID NO: 19, LCDR3 comprises an amino acid sequence according to SEQ ID NO: 6, optionally wherein one or more of the CDRs comprises one, two or three amino acid mutations relative to the recited sequences; and b) the one or more payload moieties is covalently linked to the binding molecule, optionally via one or
  • a binding molecule-payload conjugate comprising a binding molecule and one or more payload moieties; wherein: P129624PCT a) the binding molecule is an antibody or a fragment thereof and comprises heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3 according to the Chothia numbering scheme, wherein: 5 HCDR1 comprises an amino acid sequence according to SEQ ID NO: 20, HCDR2 comprises an amino acid sequence according to SEQ ID NO: 21, HCDR3 comprises an amino acid sequence according to SEQ ID NO: 17, LCDR1 comprises an amino acid sequence according to SEQ ID NO: 18, LCDR2 comprises an amino acid sequence according to SEQ ID NO: 19, LCDR3 comprises an amino acid sequence 10 according to SEQ ID NO: 6, optionally wherein one or more of the CDRs comprises one, two or three amino acid mutations relative to the recited sequences; and b) the one or more payload moieties is covalently linked to the binding
  • the Fc region is a modified Fc region. 8.
  • the Fc region comprises an amino acid substitution at position 234, or an amino acid substitution at position 235, or an amino acid substitution at position 236, or an amino acid substitution at positions 234 and 235, or an amino acid substitution at positions 234, 235 and 236, according to the EU numbering scheme. 30 10.
  • ADC antibody drug conjugate
  • the BPC according to embodiment 11, wherein the heavy chain comprises an amino acid sequence according to SEQ ID NO: 10, or a variant having at least 80% sequence identity thereto. 10 13.
  • the BPC according to embodiment 11, wherein the heavy chain comprises an amino acid sequence according to SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13 or SEQ ID NO: 14, or a variant having at least 80% sequence identity thereto.
  • the BPC according to any one of the preceding embodiments, wherein the one or more payload moieties is covalently linked to the binding molecule via a linker. 16.
  • the linker comprises or is a peptide linker. 20 17.
  • the linker comprises a first conjugation moiety for coupling with the binding molecule. 18.
  • the linker comprises a second conjugation moiety for coupling with the payload moiety. 19.
  • linker 25 comprises a peptide linker selected from AA 1 , AA 1 -Gly, Val-Cit, Val-Ala, Val-AA 1 , Val-AA 1 -Gly, AA 1 -Ala-Asn, Ala-Ala-Ala, Ala-Ala-Asn and Gly-Gly-Phe-Gly.
  • P129624PCT 20 The BPC according to embodiment 19, wherein the amino acid residue represented 5 21.
  • the BPC according to any one of embodiments 17 to 20, wherein the first conjugation moiety comprises a pyrimidine-sulfone moiety. 22.
  • the payload moiety is selected from a drug, a detectable marker, a radioisotope, a fluorescent agent, a luminescent P129624PCT agent, a colored agent, an enzyme, polyethylene glycol, a nuclide, a nucleic acid, a small molecule toxin, a polypeptide having binding activity, a protein, a receptor, a ligand, another active agent that inhibits tumor cell growth, promotes tumor cell apoptosis or necrosis. 27.
  • the BPC according to embodiment 28, wherein the cytotoxic drug is a tubulin inhibitor, a DNA damaging agent, a TOPO1 inhibitor, an auristatin, a maytansinoid, or a calicheamicin.
  • 30. The BPC according to embodiment 29, wherein the cytotoxic drug is a TOPO1 10 inhibitor.
  • the TOPO1 inhibitor is a camptothecin or an exatecan. 32.
  • the payload-linker comprises a structure shown as formula (III-C): 15 or a tautomer, a mesomer, a racemate, an enantiomer or a diastereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein L is -L a -L b -L c -; -L a - is selected from the group consisting of: 20 P129624PCT wherein wn is selected from the group consisting of integers ⁇ 0, and 0 or at least 1 methylene unit of W is independently replaced by -Cyr-, -N(R wx )C(O)-, - C(O)N(R wx )-, -C(O)-, -OC(O)-, -C(O)O-, -NR wx -, -O-, -S-, -SO-, -SO 2
  • the BPC according to embodiment 32, wherein the payload moiety of the conjugate is selected from the group consisting of the following structures: P129624PCT III-A-11 III-A-12 III-A-13 III-A-14 III-A-15 III-A-16 III-A-17. 43.
  • the BPC according to embodiment 32, wherein the payload moiety of the conjugate has the following structure: . 5 44.
  • the BPC according to embodiment 33, wherein the payload-linker moiety of the conjugate has the following structure: . 45.
  • the BPC according to embodiment 34 wherein the conjugate is selected from the group consisting of the following structural formulae: P129624PCT BM BM BM O N O H O NH O N F N O HO O q 130 P129624PCT BM B HO O q O O BM H O H N N N N N O H O H O O N O S H NH O N F N O HO O q 131 P129624PCT P129624PCT or a pharmaceutically acceptable salt thereof, wherein, q represents a connection number, and q is selected from the group consisting of integers from 1 to 16, and 5 BM is a binding molecule as described in any one of embodiments 1 to 8. 46.
  • the BPC according to embodiment 45 wherein the conjugate is: or a pharmaceutically acceptable salt thereof, wherein, q represents a connection number, and q is selected from the group consisting of integers 10 from 1 to 16. 47.
  • the BPC according to embodiment 46 wherein q is an integer from 4 to 12. 48.
  • the BPC according to embodiment 47 wherein q is an integer from 6 to 10. 49.
  • the BPC according to embodiment 48 wherein q is an integer from 7 to 9. P129624PCT 50.
  • the BPC according to any one of embodiments 1 to 31, wherein the BPC comprises a structure shown as formula (II-A): 5 or a tautomer, a mesomer, a racemate, an enantiomer or a diastereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, X 1 is saturated C, and X 1 is substituted with R n ;
  • ring A is selected from the group consisting of: 3-10 membered saturated or partially unsaturated heterocyclyl and 3-10 membered saturated or partially unsaturated 10 carbocyclyl, wherein ring A is substituted with 0 or at least 1 substituent R 1a ; when ring A is 3-10 membered saturated or partially unsaturated carbocyclyl, ring A is substituted with p L 2 , and L 2 is not R n ; or, when ring A is 3-10 membered saturated or partially unsaturated heterocyclyl, ring A is substituted with p L 2 ; 15
  • the BPC according to any one of embodiments 1 to 31, wherein the BPC comprises 15 a structure shown as formula (II-C x ): , or a tautomer, a mesomer, a racemate, an enantiomer or a diastereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein L is -L a -L b -L c -; 20 -L a - is selected from the group consisting of: P129624PCT wherein W is -(C(R wa )(R wb ))wn-, Y is -(OCH2CH2)yn-Oyp-, and Z is -(C(R za )(R zb ))zn; wherein wn is selected from the group consisting of integers ⁇ 0, and 0 or at least 1 methylene unit of W is independently replaced by -Cyr-, -N(R wx )C(O)
  • BPC comprises a structure shown as formula (II-D x ): , 10 wherein BM is the binding molecule, q is the connection number and is an integer from 1 to 16; L is -L a -L b -L c -; -L a - is selected from the group consisting of: P129624PCT wherein W is -(C(R wa )(R wb )) wn -, Y is -(OCH 2 CH 2 ) yn -O yp -, and Z is -(C(R za )(R zb )) zn ; wherein wn is selected from the group consisting of integers ⁇ 0, and 0 or at least 1 methylene unit of W is independently replaced by -Cyr-, -N(R wx )C(O)-, - C(O)N(R wx
  • -L c - is selected from the group consisting of: 15 wherein R L1 and R L2 are each independently selected from the group consisting of: hydrogen, protium, deuterium, tritium, halogen, -NO 2 , -CN, -OH, -SH, -NH 2 , -C(O)H, -CO 2 H, - C(O)C(O)H, -C(O)CH 2 C(O)H, -S(O)H, -S(O) 2 H, -C(O)NH 2 , -SO 2 NH 2 , -OC(O)H, - N(H)SO 2 H and a C 1-6 aliphatic group.
  • P129624PCT 61 The BPC according to embodiment 59, wherein -L c - is , wherein R L1 and R L2 are each independently selected from the group consisting of: hydrogen, 5 halogen, -OH and a C 1-6 aliphatic group. 62.
  • ring A is selected from the group consisting of: 3-10 membered saturated heterocyclyl and 3-10 membered saturated carbocyclyl. 67.
  • the linker-payload comprises a structure shown as formula (II-Ax): wherein, when ring A is 3-10 membered saturated or partially unsaturated carbocyclyl, ring A 15 is substituted with p L 2 , and L 2 is not R n ; or, when ring A is 3-10 membered saturated or partially unsaturated heterocyclyl, ring A is substituted with p L 2 .
  • P129624PCT 78 The BPC according to any one of embodiments 51 to 76, wherein m is 0, and L 3 is a 20 covalent bond.
  • the BPC according to embodiment 51, wherein the payload is selected from the group consisting of structures: P129624PCT II-A-9 II-A-10 II-A-11 II-A-12. 95.
  • the BPC according to embodiment 94 wherein the payload comprises the structure 96.
  • the BPC according to embodiment 95 wherein the payload has the structure P129624PCT .
  • the BPC according to embodiment 52, wherein the payload-linker moiety has the structure . 5 98.
  • the BPC according to any one of embodiments 1 to 31, wherein the conjugate has the structure represented by formula I: or a tautomer, a mesomer, a racemate, an enantiomer or a diastereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, 10 BM is the binding molecule; P129624PCT 5 each Z is independently selected from a direct bond, a carbon-carbon triple bond, a carbon- carbon double bond, and amido (preferably selected from a direct bond, a carbon- carbon triple bond, and a carbon-carbon double bond); Rx and Ry are each independently selected from H and C1-4 alkyl; each m is independently selected from 0, 1, 2, 3, 4, 5 and 6; 10 y1 is selected from any integer between 1 and 6 (such as 4, 5, and 6); each y2 is independently selected from any integer between 0 and 15 (such as 6-15); each y3 is independently selected from 1, 2, and 3; each y4 is independently selected from 0 and 1; position 1 is attached to
  • R 15 r, r 1 , r 1a and r 1b are each independently 0, 1, 2, 3, 4 or 5;
  • R m1 , R n1 , R m1a , R n1a , R m1b and R n1b are each independently H, C 1-6 alkyl or -COOR x1 , wherein, R x1 is C 1-6 alkyl; or, R m1 and R n1 , R m1a and R n1a , and R m1b and R n1b , together with the nitrogen atom to which they are both attached, form a 5-6 membered heterocyclic ring, and of said 5-6 20 membered heterocyclic ring, the heteroatom is selected from 1 or 2 N atoms; the said 5-6 membered heterocyclic ring is optionally substituted with one or more R 0’ ;
  • R z is selected from C 1-6 alkyl;
  • R 0 and R 0’ are
  • P129624PCT 5 100 The BPC according to embodiment 99, wherein L 1 is , 101.
  • 102 The BPC according to embodiment 101, wherein Z is a carbon-carbon triple bond.
  • 103 The BPC according to any one of embodiments 98 to 102, wherein Rx is H or methyl. 10 104.
  • the BPC according to embodiment 110 wherein L 3 is selected from AA 1 , AA 1 -Gly, Val- 20 Cit, Val-AA 1 -Gly, AA 1 -Ala-Asn and Gly-Gly-Phe-Gly. P129624PCT 112.
  • An antibody comprising: a) heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3 according to the IMGT numbering 10 scheme, wherein: HCDR1 comprises an amino acid sequence according to SEQ ID NO: 1, HCDR2 comprises an amino acid sequence according to SEQ ID NO: 2, HCDR3 comprises an amino acid sequence according to SEQ ID NO: 3, LCDR1 comprises an amino acid sequence according to SEQ ID NO: 4, LCDR2 comprises an amino acid sequence 15 according to SEQ ID NO: 5, LCDR3 comprises an amino acid sequence according to SEQ ID NO: 6, optionally wherein one or more of the CDRs comprises one, two or three amino acid mutations relative to the recited sequences; and b) a modified Fc region.
  • HCDR1 comprises an amino acid sequence according to SEQ ID NO: 1
  • HCDR2 comprises an amino acid sequence according to SEQ ID NO: 2
  • HCDR3 comprises an amino acid sequence according to SEQ ID NO: 3
  • An antibody comprising: 20 a) heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3 according to the Kabat numbering scheme, wherein: HCDR1 comprises an amino acid sequence according to SEQ ID NO: 15, HCDR2 comprises an amino acid sequence according to SEQ ID NO: 16, HCDR3 comprises 25 an amino acid sequence according to SEQ ID NO: 17, LCDR1 comprises an amino acid sequence according to SEQ ID NO: 18, LCDR2 comprises an amino acid sequence according to SEQ ID NO: 19, LCDR3 comprises an amino acid sequence according to SEQ ID NO: 6, optionally wherein one or more of the CDRs comprises one, two or three amino acid mutations relative to the recited sequences; and 30 b) a modified Fc region.
  • HCDR1 comprises an amino acid sequence according to SEQ ID NO: 15
  • HCDR2 comprises an amino acid sequence according to SEQ ID NO: 16
  • HCDR3 comprises 25 an amino acid sequence according to SEQ ID NO: 17
  • HCDRs heavy chain complementarity determining regions
  • LCDRs light chain complementarity determining regions
  • HCDR1 comprises an amino acid sequence according to SEQ ID NO: 20
  • HCDR2 5 comprises an amino acid sequence according to SEQ ID NO: 21
  • HCDR3 comprises an amino acid sequence according to SEQ ID NO: 17
  • LCDR1 comprises an amino acid sequence according to SEQ ID NO: 18
  • LCDR2 comprises an amino acid sequence according to SEQ ID NO: 19
  • LCDR3 comprises an amino acid sequence according to SEQ ID NO: 6, optionally wherein one or more of the CDRs comprises 10 one, two or three amino acid mutations relative to the recited sequences
  • b) a modified Fc region is optionally wherein one or more of the CDRs comprises 10 one, two or three amino acid mutations relative to the recited sequences.
  • VH heavy chain variable region
  • VL light chain variable region
  • the antibody according to embodiment 137 wherein the one or more amino acid substitution(s) are selected from the group consisting of: a) L234A, L235A, A327G, A330S, and P331S (LALA- ⁇ A); b) L234A, L235A, and K322A (LALA-KA); c) L234S, L235T, and G236R (STR); and 30 d) L234A and L235A (LALA).
  • the one or more amino acid substitution(s) are selected from the group consisting of: a) L234A, L235A, A327G, A330S, and P331S (LALA- ⁇ A); b) L234A, L235A, and K322A (LALA-KA); c) L234S, L235T, and G236R (STR); and 30 d) L234A and L235A (LALA).
  • the one or more amino acid substitution(s) are selected from the group consisting
  • the linker-payload compound is of 10 the general formula (II-Fx): , wherein, L x is Lax-Lb-Lc-; L ax - is selected from the group consisting of: 15 wherein R hal is iodine or bromine; wherein W is -(C(R wa )(R wb )) wn -, Y is -(OCH 2 CH 2 ) yn -O yp -, and Z is -(C(R za )(R zb )) zn ; P129624PCT wherein wn is selected from the group consisting of integers ⁇ 0, and 0 or at least 1 methylene unit of W is independently replaced by -
  • linker-payload compound is of the general formula (III-F): , wherein, L x is L ax -L b -L c -; 15 L ax - is selected from the group consisting of: wherein R hal is iodine or bromine; P129624PCT wherein W is -(C(R wa )(R wb )) wn -, Y is -(OCH 2 CH 2 ) yn -O yp , and Z is -(C(R za )(R zb )) zn ; wherein wn is selected from the group consisting of integers ⁇ 0, and 0 or at least 1 methylene unit of W is independently replaced by -Cyr-, -N(R wx )C(O)-, - C(O)N(R wx )-, -C(O)-,
  • the linker-payload compound is of the formula P129624PCT wherein, LG is a leaving group; 5 each Z is independently selected from a direct bond, a carbon-carbon triple bond, a carbon- carbon double bond, and amido (preferably selected from a direct bond, a carbon- 10 carbon triple bond, and a carbon-carbon double bond); Rx and Ry are each independently selected from H and C1-4 alkyl; each m is independently selected from 0, 1, 2, 3, 4, 5 and 6; y1 is selected from any integer between 1 and 6 (such as 4, 5, and 6); each y2 is independently selected from any integer between 0 and 15 (such as 6-15); 15 each y3 is independently selected from 1, 2, and 3; P129624PCT each y4 is independently selected from 0 and 1; position 1 is attached to LG, and position 2 is attached to L 2 or L 3 ; 5 and 6 (such as 4, 5, and 6); each y2 is independently selected from any integer between 0 and 10 (such as 6-10); each
  • R m1 , R n1 , R m1a , R n1a , R m1b and R n1b are each independently H, C 1-6 alkyl or -COOR x1 , wherein, R x1 is C 1-6 alkyl; P129624PCT or, R m1 and R n1 , R m1a and R n1a , and R m1b and R n1b , together with the nitrogen atom to which they are both attached, form a 5-6 membered heterocyclic ring, and of said 5-6 membered heterocyclic ring, the heteroatom is selected from 1 or 2 N atoms; the said 5-6 membered heterocyclic ring is optionally substituted with one or more R 0’ ; 5 R z is selected from C 1-6 alkyl; R 0 and R 0’ are each independently
  • the linker-payload compound is of the formula: 153.
  • a cell comprising the one or more nucleic acid sequence(s) according to embodiment 20 153, the vector according to embodiment 154, the binding molecule as defined in any one of embodiments 1 to 14, the BPC according to any one of embodiments 1 to 130, or the antibody P129624PCT according to any one of embodiments 131 to 140; optionally, wherein the cell is capable of expressing the binding molecule as defined in any one of embodiments 1 to 14 or the antibody according to any one of embodiments 131 to 140.
  • a composition comprising the BPC according to any one of embodiments 1 to 130 or 5 the antibody according to any one of embodiments 131 to 140, together with a pharmaceutically acceptable carrier, diluent or excipient. 157.
  • An in vitro method comprising contacting a cell with the BPC according to any one of embodiments 1 to 130 or the antibody according to any one of embodiments 131 to 140.
  • a method of treating or diagnosing a disease comprising administering the BPC 10 according to any one of embodiments 1 to 130, the antibody according to any one of embodiments 131 to 140, or the composition according to embodiment 156 to a subject.
  • PDAC pancreatic ductal adenocarcinoma
  • a kit comprising a BPC according to any one of embodiments 1 to 130, an antibody according to any one of embodiments 131 to 140, or a composition according to embodiment 30 156. 165.
  • An ADC comprising a binding molecule and one or more payload-linker moieties; P129624PCT wherein: a) the binding molecule is an antibody comprising a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence according to SEQ ID NO: 10 and the light chain comprises an amino acid sequence according to SEQ ID NO: 9; 5 and b) the one or more payload-linker moieties have the structure: 166.
  • An ADC comprising a binding molecule and one or more payload-linker moieties; wherein: 10 a) the binding molecule is an antibody comprising a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence according to SEQ ID NO: 13 and the light chain comprises an amino acid sequence according to SEQ ID NO: 9; and b) the one or more payload-linker moieties have the structure: 15 . 167.
  • An ADC comprising a binding molecule and one or more payload-linker moieties; wherein: a) the binding molecule is an antibody comprising a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence according to SEQ ID NO: P129624PCT 10 and the light chain comprises an amino acid sequence according to SEQ ID NO: 9; and b) the one or more payload-linker moieties have the structure: . 5 168.
  • An ADC comprising a binding molecule and one or more payload-linker moieties; wherein: a) the binding molecule is an antibody comprising a heavy chain and a light chain, wherein the heavy chain comprises an amino acid sequence according to SEQ ID NO: 10 and the light chain comprises an amino acid sequence according to SEQ ID NO: 9; 10 and b) the one or more payload-linker moieties have the structure: . 169.
  • a binding molecule-payload conjugate comprising a binding molecule and one or more payload moieties; wherein: P129624PCT a) the binding molecule is an antibody or a fragment thereof and comprises heavy chain complementarity determining regions (HCDRs) 1-3 and light chain complementarity determining regions (LCDRs) 1-3 according to the IMGT numbering scheme, wherein the binding molecule specifically binds to CA19-9; and 5 b) the one or more payload moieties is covalently linked to the binding molecule, optionally via one or more linkers. 170.
  • HCDRs 1-3 and/or LCDRs 1-3 is selected from: a) HCDR1 comprising an amino acid sequence according to SEQ ID NO: 1, 10 b) HCDR2 comprising an amino acid sequence according to SEQ ID NO: 2, c) HCDR3 comprising an amino acid sequence according to SEQ ID NO: 3, d) LCDR1 comprising an amino acid sequence according to SEQ ID NO: 4, e) LCDR2 comprising an amino acid sequence according to SEQ ID NO: 5, and f) LCDR3 comprisng an amino acid sequence according to SEQ ID NO: 6; 15 optionally wherein the at least one of HCDRs 1-3 and/or LCDRs 1-3 comprises one, two or three amino acid mutations relative to the recited sequences.

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Abstract

L'invention concerne des conjugués molécule de liaison-charge utile (BPC) comprenant une molécule de liaison et une ou plusieurs fractions de charge utile, la molécule de liaison se liant spécifiquement à CA19-9, destinés par exemple à être utilisés dans le traitement d'un adénocarcinome canalaire pancréatique (PDAC).
PCT/EP2025/065686 2024-06-06 2025-06-05 Molécule de liaison à ca19-9 Pending WO2025252894A1 (fr)

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WO2020081988A1 (fr) * 2018-10-19 2020-04-23 Memorial Sloan-Kettering Cancer Center Récepteurs antigéniques chimériques ciblant sialyl lewis a et leurs utilisations
WO2020219287A1 (fr) * 2019-04-26 2020-10-29 Immunogen, Inc. Dérivés de camptothécine
WO2022198232A1 (fr) * 2021-03-18 2022-09-22 Seagen Inc. Libération sélective de médicament à partir de conjugués internalisés de composés biologiquement actifs
WO2022200498A1 (fr) * 2021-03-26 2022-09-29 BioNTech SE Polythérapie avec un anticorps anti-ca19-9 et folfirinox dans le traitement du cancer

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