Antibodies and antibody-drug-conjugates targeting mt1-mmp

Abstract

The present disclosure relates to antibodies and molecular conjugates targeting Membrane type 1-matrix metalloproteinase (MT1-MMP), in particular antibody-drug conjugates (ADCs) of said antibodies and their use in delivery of active agents to cells and tissues expressing MT1-MMP. The invention further relates to the use of said ADCs in the treatment of cancers, such as cancers characterised by MT1-MMP expressing cells.

Description

[0001] P7363PC00

[0002] Antibodies and antibody-drug-conjugates targeting MT1-MMP

[0003] Technical field

[0004] The present disclosure relates to antibodies and molecular conjugates targeting Membrane type 1 -matrix metalloproteinase (MT1 -MMP), in particular antibody-drug conjugates (ADCs) of said antibodies and their use in delivery of active agents to cells and tissues expressing MT1 -MMP. The invention further relates to the use of said ADCs in the treatment of cancers, such as cancers characterised by MT1-MMP expressing cells.

[0005] Background

[0006] Membrane type 1 -matrix metalloproteinase (MT1 -MMP), also known as matrix metalloproteinase-14 (MMP14), is a cell surface collagenase member of the matrix metalloproteinases family. It is involved in extracellular matrix (ECM) turnover during tissue remodeling, particularly the degradation of collagen. In cancer, MT1-MMP plays a crucial role in cell-ECM interactions, influencing ECM degradation, remodeling, cell invasion, and metastasis (Turunen et al., 2017). MT1 -MMP is highly expressed in various cancers, including melanoma, pancreatic adenocarcinoma, and head and neck carcinoma, all of them with a high need for novel treatment options. Among the cancers studied, sarcomas exhibit the highest levels of MT1 -MMP expression (Gonzalez-Molina et al., 2019). MT1 -MMP is upregulated not only on malignant tumor cells but also on cancer-associated fibroblasts (CAFs), macrophages, and endothelial cells (Chun et al., 2004).

[0007] Antibody-drug conjugates (ADCs) are a rapidly advancing class of targeted therapies that have changed the treatment of various advanced-stage cancers. ADCs are complex molecules composed of an antibody (a whole mAb or an antibody fragment) linked, via a stable chemical linker, to a biologically active drug or cytotoxic compound. This design combines the specific targeting capabilities of monoclonal antibodies with the potent efficacy of cytotoxic agents, delivering the payload directly to targetexpressing tumors and inducing cell death. This means that, in contrast to traditional chemotherapy, ADCs actively attack cancer cells so that healthy cells with little or no antigen expression are less severely affected. This approach potentially broadens the therapeutic index of the cytotoxic agent by reducing systemic drug exposure and minimizing dose-limiting toxicities to off-target healthy tissues (Metrangolo & Engelholm, 2024). To date, fifteen ADCs have received market approval, with P7363PC00 hundreds more in advanced clinical and preclinical trials (Dumontet et al., 2023; Metrangolo & Engelholm, 2024).

[0008] Various means of therapy are available for most cancer types. However, in many cases they present low efficiency or detrimental side effects due to the lack of specificity. Therefore, there is a need for more efficient treatments with increased specificity.

[0009] Summary

[0010] The present disclosure relates to anti-MT1 -MMP monoclonal antibodies, in particular an anti-MT1 -MMP monoclonal antibody, designated mAb-2. The present disclosure further relates to these anti-MT1-MMP monoclonal antibodies, such as mAb-2, conjugated to one or more cytotoxic payloads, constructs known as antibody-drug conjugates (ADCs). The present inventors show here that after coupling to one or more cytotoxic payloads to the antibody, the resulting ADC is highly efficient against MT1 - MMP expressing cancer cells. This is shown by in vitro testing in high MT 1 -MMP- expressing cell lines and subsequently in an in vivo experiment in tumor-bearing mice. The invention is exemplified herein with specific payloads and coupling methods but the invention is not limited to these examples.

[0011] The antibodies and / or antibody-drug conjugates of the present disclosure may induce cell death of cells expressing MT1 -MMP, such as cancer cells, cancer-associated cells, such as cancer-associated fibroblasts (CAFs), tumour cells and / or tumour-associated cells expressing MT1 -MMP. The antibodies and / or antibody-drug conjugates of the present disclosure may inhibit the growth and / or proliferation of cells expressing MT1 - MMP, such as cancer cells, cancer-associated cells, such as cancer-associated fibroblasts (CAFs), tumour cells and / or tumour-associated cells expressing MT1-MMP.

[0012] The antibodies and / or antibody-drug conjugates of the present disclosure may have improved therapeutic properties, such as increased affinity, increased avidity, increased stability, reduced immunogenicity, increased potency, increased specificity and / or reduced off-target effect.

[0013] The antibodies and / or antibody-drug conjugates of the present disclosure may reduce or prevent tumor growth. The antibodies and / or antibody-drug conjugates of the present disclosure may reduce or prevent tumor regrowth. The antibodies and / or antibody-drug conjugates of the present disclosure may reduce tumor size. P7363PC00

[0014] The antibodies and / or antibody-drug conjugates of the present disclosure may prolong survival time of a subject suffering from cancer or having a tumor. The antibodies and / or antibody-drug conjugates of the present disclosure may increase the number of subjects suffering from cancer or having a tumor that survive.

[0015] One aspect of the present disclosure relates to an antibody or antigen-binding fragment thereof which binds to Membrane type 1 -matrix metalloproteinase (MT1 -MMP), wherein said antibody is: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 2, as CDR2 an amino acid sequence of SEQ ID NO: 3, and as CDR3 and amino acid sequence of SEQ ID NO: 4, and ii. an immunoglobulin heavy chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 6, as CDR2 an amino acid sequence of SEQ ID NO: 7, and as CDR3 and amino acid sequence of SEQ ID NO: 8, or b. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 1 or a sequence having at least 90% sequence identity thereto, and ii. an immunoglobulin heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 5 or a sequence having at least 90% sequence identity thereto, or c. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 or a sequence having at least 90% sequence identity thereto, and ii. an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27 or a sequence having at least 90% sequence identity thereto, or P7363PC00 d. a humanized version of the antibody or antigen-binding fragment thereof of any one of a, b or c, or e. a chimeric version of the antibody or antigen-binding fragment thereof of any one of a, b or c.

[0016] Another aspect of the present disclosure relates to an antibody-drug conjugate (ADC) comprising: a. the antibody or antigen-binding fragment thereof according to the present disclosure, b. an active agent, and c. optionally a linker which links a) to b).

[0017] Another aspect of the present disclosure relates to the antibody or antigen-binding fragment thereof or the antibody-drug conjugate according to the present disclosure for use as a medicament.

[0018] Another aspect of the present disclosure relates to a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof or the antibody-drug conjugate according to any the present disclosure and a pharmaceutically acceptable buffer, diluent, carrier, adjuvant or excipient.

[0019] Another aspect of the present disclosure relates to the antibody or antigen-binding fragment thereof, the antibody-drug conjugate or the pharmaceutical composition according to any the present disclosure for use in the treatment of a cancer, such as a cancer characterised by cells expressing MT1 -MMP, such as a cancer characterised by cancer cells, cancer-associated cells, such as cancer-associated fibroblasts (CAFs), tumour cells and / or tumour-associated cells expressing MT1 -MMP.

[0020] Another aspect of the present disclosure relates to an isolated polynucleotide comprising a nucleotide sequence encoding an amino acid sequence comprising the antibody or antigen-binding fragment thereof according to the present disclosure.

[0021] Another aspect of the present disclosure relates to a vector, such as an expression vector, comprising the isolated polynucleotide according to the present disclosure. P7363PC00

[0022] Another aspect of the present disclosure relates to a host cell, such as a recombinant host cell, comprising the isolated polynucleotide and / or the vector according to the present disclosure.

[0023] Description of Figures

[0024] Fig. 1. Structure of an antibody-drug conjugate (ADC). A. An ADC comprises a tumor-targeting monoclonal antibody (mAb) chemically linked to a cytotoxic payload via a linker. Most ADC payloads fall into two categories: anti-mitotic or DNA damaging cytotoxins. Anti-mitotic payloads (e.g., monomethyl auristatin E (MMAE)) interfere with the tubulin-polymerization dynamics and induce mitotic arrest, blocking rapid tumor proliferation, while DNA-damaging payloads (e.g., PNU-159682) inhibit cell proliferation via irreversible DNA damage. B. The examples shown are the cysteine and glycan conjugation methods. The PNU-containing ADC used in the examples is developed using the conserved N- glycosylation at Asn297 and click chemistry, resulting in two molecules of drug per mAb. The MMAE-containing ADC used in the examples is constructed through the cysteine conjugation, which uses the interchain cysteines and maleimide chemistry to conjugate the antibody to the payload, resulting in approx. 4 cytotoxin molecules per mAb.

[0025] Fig. 2. In vitro cell viability in HT1080 fibrosarcoma cell line after ADC treatment. A) Effect of mAb-2-PNU ADC in comparison with a non-targeted control ADC (HyHEL- PNU). B) Effect of mAb-2-PNU in comparison with an alternative non-targeted control ADC (aTNP-PNU). C) Effect of mAb-2-MMAE ADC in comparison with a non-targeted control ADC (HyHEL-MMAE). D) Effect of mAb-2-PNU in comparison with mAb-2- MMAE. In A and B, ADCs were added at a maximum concentration of 5 pg / ml and serial-diluted tenfold. In C and D, ADCs were added at a maximum concentration of 15 pig / ml and serial-diluted fivefold. The non-binding controls aTNP-PNU and HyHEL-PNU ADCs were included for comparison. It can be concluded that both mAb-2- PNU and mAb-2-MMAE reduce HT1080 cell viability notably in comparison with both control ADCs.

[0026] Fig. 3. In vitro cell viability in two different osteosarcoma cell lines (MG-63 and SaOS-2). A) MG-63 viability. Effect of mAb-2-PNU ADC in comparison with a nontargeted control ADC (HyHEL-PNU). B) MG-63 viability. Effect of mAb-2-PNU in comparison with a non-targeted control ADC (aTNP-PNU). C) SaOS-2 viability. Effect P7363PC00 of mAb-2-PNU ADC in comparison with a non-targeted control ADC (HyHEL-PNU). D) SaOS-2 viability. Effect of mAb-2-PNU in comparison with a non-targeted control ADC (aTNP-PNU). The ADCs were added at a maximum concentration of 15pg / ml and serial-diluted tenfold. The non-binding controls aTNP-PNU and HyHEL-PNU ADCs were included for comparison. It can be concluded that both mAb-2-PNU reduce both MG-63 and SaOS-2 cell viability notably in comparison with both control ADCs.

[0027] Fig. 4. Effect on in vitro cell viability of different anti-MT1-MMP1-PNU ADCs in HT1080 fibrosarcoma cell line. The effect of mAb-2-PNU on cell viability is compared with that of ADCs based on other antibodies against MT1-MMP. A) Effect of mAb-2- PNU in comparison with HyHEL-PNU non-targeted control ADC. B) Effect of mAb-1- PNU in comparison with HyHEL-PNU non-targeted control ADC. C) Effect of 9E8:A7- PNU in comparison with HyHEL-PNU non-targeted control ADC. The ADCs were added at a maximum concentration of 5pg / ml and serial-diluted fourfold. It can be concluded that all of the three anti-MT1-MMP ADCs have an effect in comparison with the control ADC. However, mAb-2-PNU (A) reduces cell viability at a concentration 10- times lower than the other ADCs, thus, mAb-2-PNU presents higher potency than ACDs comprising known antibodies against MT1 -MMP.

[0028] Fig. 5. In vivo treatment of tumors: Mice with subcutaneous HT1080 tumors subjected to tail-vein administration of mAb-2-PNU or control reagents. Mice were inoculated with HT1080 cells by subcutaneous injection. Upon the appearance of palpable tumors, local treatment with the indicated reagents was initiated by tail-vein injection. Treatments were given as doses of 0.75 mg / kg at day 0, 7, and 14 (black arrows). Treatment groups received PBS (N=8), non-targeted control ADC HyHEL- PNU (N=9) or mAb-2-PNU (N=9). A) Tumor size with individual values for each treatment group up to day 100. Complete tumor regression is observed in the mAb-2- PNU group. B) Mouse survival curve. The graph represents the survival percentage for each treatment group. Whereas all mice treated with PBS or HyHEL-PNU had consistent tumor growth, reaching the point of sacrifice (ethically predecided maximum tumor burden) before day 26, 7 out of 9 mice treated with mAb-2-PNU survived and showed no tumor regrowth throughout the experiment (termination at day 100).

[0029] Fig. 6. In vitro viability of MDA-MB-231 breast carcinoma cells in the presence of mAb-2-based ADCs with different payloads. A) Effect of mAb-2-PNU in comparison P7363PC00 with HyHEL-PNU non-targeted control ADC. B) Effect of mAb-2-MMAE in comparison with HyHEL-MMAE non-targeted control ADC. C) Effect of mAb-2-PNU in comparison with mAb-2-MMAE. The PNU ADCs were added at a maximum concentration of A) 10 pig / ml and serial-diluted five-fold or C) 5 pg / ml and serial-diluted fourfold. B and C) MMAE ADCs were added at a maximum concentration of 35 pg / ml and serial-diluted fivefold. It can be concluded that both mAb-2-PNU and mAb-2-MMAE are effective in comparison with the non-targeted control ADC.

[0030] Fig. 7. In vitro viability of EBC-1 lung squamous cell carcinoma cells in the presence of mAb-2-based ADCs with different payloads. A) Effect of mAb-2-PNU in comparison with HyHEL-PNU non-targeted control ADC. B) Effect of mAb-2-MMAE in comparison with HyHEL-MMAE non-targeted control ADC. C) Effect of mAb-2-PNU in comparison with mAb-2-MMAE. The PNU ADCs were added at a maximum concentration of A) 10 pg / ml and serial-diluted five-fold or C) 5 pg / ml and serial-diluted fourfold. B and C) MMAE ADCs were added at a maximum concentration of 35 pg / ml and serial-diluted fivefold. It can be concluded that both mAb-2-PNU and mAb-2-MMAE are effective in comparison with the non-targeted control ADC. When comparing mAb- 2-PNU and mAb-2-MMAE, they present a very similar effect in this cell line.

[0031] Fig. 8. In vitro viability of U87-MG malignant glioma cells in the presence of mAb- 2-PNU. Effect of mAb-2-PNU in comparison with non-targeted control ADC HyHEL- PNU. The ADCs were added at a maximum concentration of 10 pg / ml and serial- diluted fivefold.

[0032] Fig. 9. Calculation of the Drug-antibody ratio (DAR). Equation used to calculate the DAR of both PNU and MMAE-based ADCs, e280corresponds to the extinction coefficient at 280 nm of the drug and the antibody, e corresponds to the extinction coefficient at A nm of the drug and the antibody, where A is the absorption spectrum maximum of the drug. Cdrug and CmAb represent their molar concentration. Modified from Chen (2014).

[0033] Fig. 10. In vitro cell viability in BxPC3 pancreatic adenocarcinoma cell line after ADC treatment. A) Effect of mAb-2-PNU ADC in comparison with a non-targeted control ADC (HyHEL-PNU). B) Effect of mAb-2-MMAE ADC in comparison with a nontargeted control ADC (HyHEL-MMAE). In both A and B, the ADCs were added at a P7363PC00 maximum concentration of 15 pg / ml and serial-diluted five-fold. It can be concluded that both mAb-2- PNU and mAb-2-MMAE effectively kill BxPC3 cells in comparison with control ADCs.

[0034] Fig. 11. In vivo treatment of tumors: Mice with subcutaneous MDA-MB-231 (breast carcinoma) tumors subjected to tail-vein administration of mAb-2-PNU or control reagents. Mice were inoculated with MDA-MB-231 cells by subcutaneous injection. Upon the appearance of palpable tumors, treatment with the indicated reagents was administered by tail-vein injection. Treatments were given as doses of 0.75 mg / kg at day 0, 7, and 14 (black arrows). Treatment groups received PBS (N=8), non-targeted control ADC HyHEL-PNU (N=8) or mAb-2-PNU (N=9). A) Tumor size with individual values for each treatment group up to day 42. Prominent regression in tumor size followed by an extended period of disease control is observed in the mAb-2-PNU group. B) Mouse survival curve. The graph represents the survival percentage for each treatment group. Whereas mice treated with PBS or HyHEL-PNU had consistent tumor growth, with 90% reaching the point of sacrifice (ethically predecided maximum tumor burden) before day 14, treatment with mAb-2-PNU resulted in prolonged survival with a median survival time of 41 days after first treatment.

[0035] Fig.12. In vivo treatment of tumors: Mice with subcutaneous EBC-1 (lung carcinoma) tumors subjected to tail-vein administration of mAb-2-PNU or control reagents. Study design as in Fig. 11 with the following modifications: PBS (N=8), nontargeted control ADC HyHEL-PNU (N=8) and mAb-2-PNU (N=8). A) Tumor size with individual values for each treatment group up to day 80. Complete tumor regression is observed in the mAb-2-PNU group until day 60. B) Mouse survival curve. The graph represents the survival percentage for each treatment group. None of the mice in the mAb-2-PNU reached the point of sacrifice due to tumor size (ethically predecided maximum) during the duration of the study, although three mice were terminated due to weight loss. PNU is a highly toxic payload and the ADC is dosed close to the max tolerated dose and it can therefore be expected that a minor fraction of the mice would suffer from side effects. The majority of the treated mice tolerated the mAb-2-PNU ADC (see also Fig. 5 and Fig.11 ). Mice treated with PBS or HyHEL-PNU had consistent tumor growth, reaching the point of sacrifice before day 21 and 32, respectively. P7363PC00

[0036] Fig. 13. In vivo treatment of tumors: Mice with subcutaneous EBC-1 (lung carcinoma) tumors subjected to tail-vein administration of mAb-2-MMAE or control reagents. Study design as in Fig. 11 with the following modifications: The ADCs were dosed at 4 mg / kg, PBS (N=8), non-targeted control ADC HyHEL-MMAE (N=7) and mAb-2-MMAE (N=8). A) Tumor size with individual values for each treatment group up to day 100. Complete tumor regression is observed in all the mAb-2-MMAE treated mice. B) Mouse survival curve. The graph represents the survival percentage for each treatment group. Treatment with mAb-2-MMAE prominently extended the median survival time of the mice with no mice in this group reaching the point of sacrifice (ethically predecided maximum tumor burden) during the follow-up period of 100 days after first treatment.

[0037] Fig. 14. In vivo treatment of tumors: Mice with subcutaneous U-87 MG (malignant glioma) tumors subjected to tail-vein administration of mAb-2-MMAE or control reagents. Study design as in Fig. 11 with the following modifications: The ADCs were dosed at 6 mg / kg, PBS (N=4), isotype control (IC) ADC IC-MMAE (N=4) and mAb-2- MMAE (N=4). A) Tumor size with individual values for each treatment group up to day 50. Treatment with mAb-2-MMAE resulted in an extended period of tumor regression and disease control. B) Mouse survival curve. The graph represents the survival percentage for each treatment group. Mice treated with mAb-2-PNU showed extended survival in comparison to mice treated with PBS or IC-MMAE.

[0038] Fig 15. Comparative analysis of cytotoxicity of mAb-2-MMAE and DX2400-MMAE. Effect of mAb-2-MMAE and DX2400-MMAE on A) lung squamous cell carcinoma cell line, EBC-1 and B) fibrosarcoma cell line, HT1080. ADCs were added at a maximum concentration of 10 pg / ml and serial-diluted three-fold. The mAb-2-MMAE showed superior effect over DX2400-MMAE, with an approximate three-fold lower ADC concentration resulting in 50% reduction in cell viability.

[0039] P7363PC00

[0040] Detailed description

[0041] Anti-MT1 -MMP antibodies

[0042] Some embodiments of the present disclosure relate to an antibody or antigen-binding fragment thereof which binds to Membrane type 1 -matrix metalloproteinase (MT 1 - MMP), wherein said antibody is: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 2, as CDR2 an amino acid sequence of SEQ ID NO: 3, and as CDR3 and amino acid sequence of SEQ ID NO: 4, and ii. an immunoglobulin heavy chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 6, as CDR2 an amino acid sequence of SEQ ID NO: 7, and as CDR3 and amino acid sequence of SEQ ID NO: 8, or b. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 1 or a sequence having at least 90% sequence identity thereto, and ii. an immunoglobulin heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 5 or a sequence having at least 90% sequence identity thereto, or c. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 or a sequence having at least 90% sequence identity thereto, and ii. an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27 or a sequence having at least 90% sequence identity thereto, or d. a humanized version of the antibody or antigen-binding fragment thereof of any one of a, b or c, or e. a chimeric version of the antibody or antigen-binding fragment thereof of any one of a, b or c. P7363PC00

[0043] In some embodiments, any sequence variance is outside the complementaritydetermining regions.

[0044] In some embodiments, the antibody or antigen-binding fragment thereof comprises: a. an immunoglobulin light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 1 and an immunoglobulin heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 5, or b. an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 and an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27.

[0045] In some embodiments, the antibody or antigen-binding fragment thereof comprises an immunoglobulin light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 1 and an immunoglobulin heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 5.

[0046] In some embodiments, the antibody or antigen-binding fragment thereof comprises an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 and an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27.

[0047] In some embodiments, the antibody is selected from a mouse antibody, a chimeric antibody, a human antibody, a humanised antibody, a humanised antigen-binding fragment, a Fab fragment, a Fab’ fragment, an F(ab’)2 fragment, an Fv, a single chain antibody (SCA) such as an scFv, the variable portion of the heavy and / or light chains thereof, and a Fab miniantibody.

[0048] In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanised or fully human monoclonal antibody or antigen-binding fragment thereof. In some embodiments, the antibody is a recombinant antibody.

[0049] In some embodiments, the antibody is an immunoglobulin class antibody such as IgG, IgM, IgD, IgE, IgA, and any subclass thereof, such as lgG1 , lgG2, lgG3 or lgG4. P7363PC00

[0050] Antibody-drug conjugates (ADCs) comprising anti-MT1-MMP antibodies

[0051] Some embodiments of the present disclosure relate to an antibody-drug conjugate (ADC) comprising: a. the antibody or antigen-binding fragment thereof according to the present disclosure, b. an active agent, and c. optionally a linker which links a) to b).

[0052] The active agent may be any active agent described in the section “Active agent” and the linker may be any linker described in the section “Linker”.

[0053] In some embodiments, the drug-to-antibody ratio (DAR) is between 1 and 10, such as between 2 and 8, such as between 2 and 6, such as between 2 and 4. In some embodiments, the drug-to-antibody ratio (DAR) is 2 or 4.

[0054] In some embodiments, the active agent and the linker comprise or consist of a molecular structure according to Formula I prior to conjugation:

[0055] Formula I.

[0056] In some embodiments, the active agent and the linker comprise or consist of a molecular structure according to Formula II prior to conjugation: P7363PC00

[0057] Formula II.

[0058] In some embodiments, the antibody-drug conjugate comprises or consists of: a. the antibody or antigen-binding fragment thereof according the present disclosure, b. valine-citrulline (VC), c. dibenzocyclooctyne-amine (DBCO), optionally N-functionalized with PEG4, d. PAB, and e. PNU-159682.

[0059] In some embodiments, the antibody-drug conjugate comprises or consists of: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 2, as CDR2 an amino acid sequence of SEQ ID NO: 3, and as CDR3 and amino acid sequence of SEQ ID NO: 4, and ii. an immunoglobulin heavy chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 6, as CDR2 an amino acid sequence of P7363PC00

[0060] SEQ ID NO: 7, and as CDR3 and amino acid sequence of SEQ ID NO: 8, b. valine-citrulline (VC), c. dibenzocyclooctyne-amine (DBCO), optionally N-functionalized with PEG4, d. PAB, and e. PNU-159682.

[0061] In some embodiments, the antibody-drug conjugate comprises or consists of: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 1 and an immunoglobulin heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 5, or ii. an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 and an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27, b. valine-citrulline (VC), c. dibenzocyclooctyne-amine (DBCO), optionally N-functionalized with PEG4, d. PAB, and e. PNU-159682.

[0062] In some embodiments, the antibody-drug conjugate comprises or consists of: a. the antibody or antigen-binding fragment thereof according the present disclosure, b. maleimidocaproyl (MC), c. valine-citrulline (VC), d. PAB, and e. monomethyl auristatin E (MMAE).

[0063] In some embodiments, the antibody-drug conjugate comprises or consists of: a. an antibody or antigen-binding fragment thereof comprising:

[0064] I . an immunoglobulin light chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence P7363PC00 of SEQ ID NO: 2, as CDR2 an amino acid sequence of SEQ ID NO: 3, and as CDR3 and amino acid sequence of SEQ ID NO: 4, and ii. an immunoglobulin heavy chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 6, as CDR2 an amino acid sequence of SEQ ID NO: 7, and as CDR3 and amino acid sequence of SEQ ID NO: 8, b. maleimidocaproyl (MC), c. valine-citrulline (VC), d. PAB, and e. monomethyl auristatin E (MMAE).

[0065] In some embodiments, the antibody-drug conjugate comprises or consists of: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 1 and an immunoglobulin heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 5, or ii. an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 and an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27, b. maleimidocaproyl (MC), c. valine-citrulline (VC), d. PAB, and e. monomethyl auristatin E (MMAE).

[0066] In some embodiments, the antibody-drug conjugate comprises or consists of: a. the antibody or antigen-binding fragment thereof according the present disclosure, b. DBCO-PEG4-VC-PAB-DMAE-PNU159682.

[0067] In some embodiments, the antibody-drug conjugate comprises or consists of: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 2, as CDR2 an amino acid sequence of P7363PC00

[0068] SEQ ID NO: 3, and as CDR3 and amino acid sequence of SEQ ID NO: 4, and ii. an immunoglobulin heavy chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 6, as CDR2 an amino acid sequence of SEQ ID NO: 7, and as CDR3 and amino acid sequence of SEQ ID NO: 8, and b. DBCO-PEG4-VC-PAB-DMAE-PNU159682.

[0069] In some embodiments, the antibody-drug conjugate comprises or consists of: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 1 and an immunoglobulin heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 5, or ii. an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 and an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27, and b. DBCO-PEG4-VC-PAB-DMAE-PNU159682.

[0070] As used herein, the term "regioisomer" or "regioisomers" refers to the positional isomers, which is a category of structural isomers, wherein the position or the substituent changes position on the parent structure. Herein the term regioisomer without departing from the chemical structure of azide-functionalized DBCO-PEG4-VC- PAB-DMEA-PNU159682 inherently includes all regioisomers either as a pure regioisomer or mixture of two or more regioisomers thereof. The regioisomers identified herein preferably refers to the following positional and / or structural isomers (I) and (II) of the DBCO-moiety: P7363PC00 wherein Ab represents the antibody or antigen-binding fragment thereof, and R represents a linking moiety connected to a cytotoxic drug, preferably PEG4-VC-PAB- DMEA-PNU159682.

[0071] In some embodiments, the antibody-drug conjugate comprises or consists of the following structure or a regioisomer thereof: wherein the wiggled line denotes any chemical bonding to the antibody.

[0072] In some embodiments, the antibody-drug conjugate comprises or consists of: a. the antibody or antigen-binding fragment thereof according the present disclosure, and b. MC-vc-PAB-MMAE.

[0073] In some embodiments, the antibody-drug conjugate comprises or consists of: P7363PC00 a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 2, as CDR2 an amino acid sequence of SEQ ID NO: 3, and as CDR3 and amino acid sequence of SEQ ID NO: 4, and ii. an immunoglobulin heavy chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 6, as CDR2 an amino acid sequence of SEQ ID NO: 7, and as CDR3 and amino acid sequence of SEQ ID NO: 8, and b. MC-vc-PAB-MMAE.

[0074] In some embodiments, the antibody-drug conjugate comprises or consists of: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 1 and an immunoglobulin heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 5, or ii. an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 and an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27, and b. MC-vc-PAB-MMAE.

[0075] In some embodiments, the antibody-drug conjugate comprises or consists of the following structure or a regioisomer thereof: P7363PC00 wherein the wiggled line denotes any chemical bonding to the antibody.

[0076] In one embodiment of the present disclosure, the antibody or the antibody-drug conjugate induces liberation of free cytotoxin from the MT1 -MMP expressing cells, leading to cell death and / or inhibition of the growth and / or proliferation of neighbouring cancer cells. This might be referred to as “bystander effect” within the present disclosure. Such as bystander effect has previously been shown to be efficient in killing cancer cells for another ADC targeting a different protein expressed by CAFs (James W. Purcell et al 2018).

[0077] Active agent

[0078] Some embodiments of the present disclosure relate to an antibody-drug conjugate (ADC) comprising: a. the antibody or antigen-binding fragment thereof according to the present disclosure, b. an active agent, and c. optionally a linker which links a) to b).

[0079] In some embodiments, the active agent is selected from a therapeutic agent, a cytotoxic agent, a radioisotope, and a detectable label. In some embodiments, the active agent is a cytotoxic agent.

[0080] In some embodiments, the active agent is a therapeutic agent, such as a therapeutic agent selected from the group consisting of anti-microtubule / anti-mitotic agents, DNA crosslinking agents, DNA alkylating agents, DNA strand scission agents, anthracyclines, antimetabolites, histone deacetylase inhibitors, kinase inhibitors, metabolism inhibitors, peptide antibiotics, immune checkpoint inhibitors, platinumbased antineoplastics, topoisomerase inhibitors, DNA or RNA polymerase inhibitors, nucleotide based agents, and cytotoxic antibiotics.

[0081] In some embodiments, the active agent is an anti-mitotic agent, such as selected from the group consisting of derivatives of auristatin or dolastatin such as monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF) and more, a taxane such as Paclitaxel or Docetaxel and more, a vinca alkaloid such as Vinblastine, Vincristine, Vindesine or Vinorelbine and more, a mayatansinoid, Colchicine, and Podophyllotoxin. P7363PC00

[0082] In some embodiments, the active agent is monomethyl auristatin E (MMAE).

[0083] In some embodiments, the active agent is a DNA-crosslinking agent, such as a DNA crosslinking agent selected from cisplatin or a derivative of cisplatin such as carboplatin or oxaliplatin, mitomycin C (MMC), pyrrolobenzodiazepine, and dimeric pyrrolobenzodiazepine derivatives such as SGD-1882.

[0084] In some embodiments, the active agent is a DNA alkylating agent, such as a DNA alkylating agent selected from nitrogen mustards such as tris(2-chloroethyl)amine, pyridinobenzodiazepines or a pyridinobenzodiazepine derivative, indolinobenzodiazepine dimers, and Duocarmycin SA.

[0085] In some embodiments, the active agent is a DNA strand scission agent, such as a DNA strand scission agent selected from calicheamicin and hamiltrone.

[0086] In some embodiments, the active agent is an anthracycline, such as an anthracycline selected from Daunorubicin, doxorubicin, epirubicin, idarubicin, and PNU-159682.

[0087] In some embodiments, the active agent is PNU-159682.

[0088] In some embodiments, the active agent is an antimetabolite, such as an antimetabolite selected from folic acid antagonists such as methotrexate, purine antimetabolites such as 6-mercaptopurine or 6-thioguanine or fludarabine phosphate or pentostatin or cladribine, and pyrimidine antimetabolites such as 5-fluorouracil or 5-fluorodeoxyuridine or cytarabine or gemcitabine.

[0089] In some embodiments, the active agent is a histone deacetylase inhibitor, such as a histone deacetylase inhibitor selected from trichostatin A, vorinostat, belinostat, panabiostat, givinostat, resminostat, abexinostat, quisinostat, rocilinostat, practinostat, CHR-3996, valproic acid, butyric acid, phenylbutyric acid, entinostat, tacedinaline, 4SC202, mocetinostat, romidepsin, nicotinamide, sirtinol, cambinol, and EX-527.

[0090] In some embodiments, the active agent is a kinase inhibitor, such as a kinase inhibitor selected from genistein, lavendustin C, PP1 -AG1872, PP2-AG1879, SU6656, P7363PC00

[0091] CGP77675, PD166285, imatinib, erlotinib, gefitinib, lavendustin A, cetuximab, UCS15A, herbimycin A, and radicicol.

[0092] In some embodiments, the active agent is a metabolism inhibitor, such as an NAMPT inhibitor selected from APO866, GMX-1777, GMX-1778 ATG-019, and OT-82.

[0093] In some embodiments, the active agent is an immune checkpoint inhibitor, such as a PD-1 inhibitor selected from Pembrolizumab, Nivolumab, Cemiplimab, JTX-4014, Spartalizumab, Camrelizumab, Sintilimab, Tislelizumab, Toripalimab, Dostarlimab, AMP-224 and AMP-514; or a PD-L1 inhibitor selected from Atezolizumab, Avelumab, Durvalumab, KN035, CK-301 , AUNP12, CA-170 and BMS-986189.

[0094] In some embodiments, the active agent is a platinum-based antineoplastic, such as a platinum-based antineoplastic selected from lipoplatin, cisplatin, carboplatin, oxaliplatin, nedaplatin, picoplatin, phenanthriplatin, satraplatin, and triplatin tetranitrate.

[0095] In some embodiments, the active agent is a topoisomerase inhibitor, such as a topoisomerase inhibitor selected from camptothecin or derivatives thereof such as topotecan, belotecan, lurtotecan, irinotecan, SN-38, exatecan, and Dxd.

[0096] In some embodiments, the active agent is a DNA- or RNA-polymerase inhibitor, such as a polymerase inhibitor selected from amanitin or alpha-amanitin or derivatives thereof, actinomycin D, and aphidicolin.

[0097] In some embodiments, the active agent comprises a radioisotope selected from60Co, 89S r, 90y, 99mTc, 131 1 , 137Cs153^an(J223Rd

[0098] Linker

[0099] Some embodiments of the present disclosure relate to an antibody-drug conjugate (ADC) comprising: a. the antibody or antigen-binding fragment thereof according to the present disclosure, b. an active agent, and c. a linker which links a) to b). P7363PC00

[0100] In some embodiments, the antibody-drug conjugate comprises a linker. In some embodiments, the antibody-drug conjugate comprises a linker selected from a cleavable and a non-cleavable linker.

[0101] In some embodiments, the antibody-drug conjugate comprises a peptide linker. In some embodiments, the linker comprises a peptide linker. In some embodiments, the linker comprises a dipeptide. In some embodiments, the linker comprises valinecitrulline (VC) and / or valine-alanine (VA).

[0102] In some embodiments, the linker comprises p-aminobenzoic acid (PAB), p- aminobenzylcarbamate (PABC), p-aminobenzoyloxycabonyl, or polyethylenglycol (PEG). In some embodiments, the linker comprises p-aminobenzoic acid (PAB).

[0103] In some embodiments, the linker comprises a dibenzocyclooctyne (DBCO) including amine-functionalized dibenzocyclooctynes (DBCO), optionally modified with a PEG, such as PEG4, maleimide, N-hydroxysuccinimide, modified or unmodified proteinbound carbohydrate, peptide sequences that are required for enzymatic reactions, azides or alkynes or being derived from these by reaction with the antibody or a chemically or enzymatically generated derivative thereof. In some embodiments, the linker comprises dibenzocyclooctyne (DBCO), such as an amine-functionalized dibenzocyclooctyne (DBCO), and PEG4.

[0104] In some embodiments, the linker comprises maleimidocaproyl (MC), optionally modified with a PEG, such as PEG4, maleimide, N-hydroxysuccinimide, modified or unmodified protein-bound carbohydrate, peptide sequences that are required for enzymatic reactions, azides or alkynes or being derived from these by reaction with the antibody or a chemically or enzymatically generated derivative thereof. In some embodiments, the linker comprises maleimidocaproyl (MC).

[0105] In some embodiments, the linker comprises a reaction product between an antibody functionalized and / or adapted with a moiety having a first specific reactivity, and a cleavable linker, such as in a linker-payload moiety, functionalized and / or adapted with a second chemical moiety having a second specific reactivity suitable for matching the first specific reactivity to engage in a chemical reaction. Examples of such matching chemical reactivity is well-known in the field of chemistry and may in one embodiment P7363PC00 take the form of alkyne / azide click chemistry, such as SPAAC or CuAAC type reactivity, exemplified by e.g., coupling of azide functionalized antibodies to linkerpayloads comprising alkyne groups, such as dibenzocyclooctyne-amine (DBCO) and / or azadibenzocyclooctyne (ADIBO). In another embodiment, this linker comprises maleimidocaproyl (sometimes abbreviated MC), where maleimide-part reacts preferably with cysteine thiols of the antibody during coupling. In other embodiments, the linker comprises N-hydroxysuccinimide, reactive attachment groups directed to modified or unmodified protein-bound carbohydrate, peptide sequences that are required for enzymatic reactions, azides or alkynes or being derived from these by reaction with the antibody or a chemically or enzymatically generated derivative thereof.

[0106] In some embodiments, SPAAC-type reaction results in one or more regioisomers, all of which are within the scope of the present disclosure.

[0107] Pharmaceutical compositions

[0108] Some embodiments of the present disclosure relate to a pharmaceutical composition comprising the antibody or antigen-binding fragment thereof or the antibody-drug conjugate according to the present disclosure and a pharmaceutically acceptable buffer, diluent, carrier, adjuvant or excipient.

[0109] Therapeutic uses

[0110] Some embodiments of the present disclosure relate the antibody or antigen-binding fragment thereof, the antibody-drug conjugate or the pharmaceutical composition according to the present disclosure for use as a medicament.

[0111] Some embodiments of the present disclosure relate the antibody or antigen-binding fragment thereof, the antibody-drug conjugate or the pharmaceutical composition according to the present disclosure for use in the treatment of a cancer, such as a cancer characterised by cells expressing MT1 -MMP, such as a cancer characterised by cancer cells, cancer-associated cells, such as cancer-associated fibroblasts (CAFs), tumour cells and / or tumour-associated cells expressing MT1 -MMP.

[0112] In some embodiments, the cells expressing MT 1 -MMP display MT 1 -MMP overexpression, optionally wherein the cells expressing MT1 -MMP are cancer cells, P7363PC00 cancer-associated cells, such as cancer-associated fibroblasts (CAFs), tumour cells and / or tumour-associated cells.

[0113] In some embodiments, the antibody, antibody-drug conjugate or composition induces cell death and / or inhibits the growth and / or proliferation of the cells expressing MT1 - MMP, such as cancer cells, cancer-associated cells, such as cancer-associated fibroblasts (CAFs), tumour cells and / or tumour-associated cells expressing MT1-MMP.

[0114] In some embodiments, the cancer is fibrosarcoma, osteosarcoma, malignant glioma, lung squamous cell carcinoma or breast carcinoma.

[0115] In some embodiments, the cancer is fibrosarcoma, osteosarcoma, malignant glioma, lung squamous cell carcinoma, breast carcinoma, pancreatic adenocarcinoma, triplenegative breast cancer or glioblastoma.

[0116] In some embodiments, the cancer is pancreatic cancer, breast cancer, colon cancer, gastric cancer, esophageal cancer, hepatocellular cancer, prostate cancer, bladder cancer, bone cancer, lung cancer, mesothelioma or squamous cell carcinomas.

[0117] In some embodiments, the cancer is pancreatic adenocarcinoma (PAAD) such as pancreatic ductal adenocarcinoma (PDAC), bladder urothelial carcinoma (BLCA), breast invasive carcinoma (BRCA), cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC), cholangiocarcinoma (CHOL), colon adenocarcinoma (COAD), esophageal carcinoma (ESCA), glioblastoma multiforme (GBM), head and neck squamous cell carcinoma (HNSCC), acute myeloid leukemia (LAML), ovarian serous cystadenocarcinoma (OV), rectum adenocarcinoma (READ), sarcoma (SARC), skin cutaneous melanoma (SKCM) or stomach adenocarcinoma (STAD).

[0118] In some embodiments, the cancer is breast cancer, such triple-negative breast cancer (TNBC), basal-like breast cancer (BLBC), HER-2 breast cancer, Luminal A type breast cancer or Luminal B type breast cancer.

[0119] In some embodiments, the cancer is metastatic cancer. P7363PC00

[0120] In some embodiments, the antibody or antigen-binding fragment thereof, the antibodydrug conjugate or the pharmaceutical composition according to the present disclosure is administered to a subject in need thereof, such as a subject suffering from cancer.

[0121] In some embodiments, administration is parenteral. In some embodiments, administration is intravenously, intracerebroventricularly, intraarticularly, intra-arterially, intraperitoneally, intrathecally, intraventricularly, intrasternally, intracranially, intramuscularly or subcutaneously or by infusion techniques. In some embodiments, administration is intravenously.

[0122] In some embodiments, the antibody or antigen-binding fragment thereof, antibody-drug conjugate or composition according the present disclosure is administered in combination with one or more further agents, such as one or more further therapeutic agents.

[0123] In some embodiments, the treatment is ameliorative or curative.

[0124] Some embodiments of the present disclosure relate to a method for treatment of a cancer, said method comprising administering to the subject the antibody or antigenbinding fragment thereof, the antibody-drug conjugate or the pharmaceutical composition according to the present disclosure.

[0125] Some embodiments of the present disclosure relate to a method for treatment of a cancer, such as a cancer characterised by cells expressing MT1 -MMP, such as a cancer characterised by cancer cells, cancer-associated cells, such as cancer- associated fibroblasts (CAFs), tumour cells and / or tumour-associated cells expressing MT1 -MMP, said method comprising administering to the subject the antibody or antigen-binding fragment thereof, the antibody-drug conjugate or the pharmaceutical composition according to the present disclosure.

[0126] Some embodiments of the present disclosure relate to a method for treatment of a cancer characterised by cells expressing MT1 -MMP, said method comprising administering to the subject the antibody or antigen-binding fragment thereof, the antibody-drug conjugate or the pharmaceutical composition according to the present disclosure. P7363PC00

[0127] Some embodiments of the present disclosure relate to a method for treatment of a cancer characterised by cancer cells, cancer-associated cells, such as cancer- associated fibroblasts (CAFs), tumour cells and / or tumour-associated cells expressing MT1 -MMP, said method comprising administering to the subject the antibody or antigen-binding fragment thereof, the antibody-drug conjugate or the pharmaceutical composition according to the present disclosure.

[0128] Some embodiments of the present disclosure relate to a method for inhibiting tumour progression in a subject, comprising administering to the subject the antibody or antigen-binding fragment thereof, the antibody-drug conjugate or the pharmaceutical composition according to the present disclosure.

[0129] Some embodiments of the present disclosure relate to a method for inhibiting, lowering or eliminating metastatic capacity of a tumour in a subject, comprising administering to the subject the antibody or antigen-binding fragment thereof, the antibody-drug conjugate or the pharmaceutical composition according to the present disclosure to said subject.

[0130] Some embodiments of the present disclosure relate to the antibody or antigen-binding fragment thereof, the antibody-drug conjugate or the pharmaceutical composition according to the present disclosure for use in a method for inhibiting tumour progression in a subject.

[0131] Some embodiments of the present disclosure relate to a kit comprising the antibody or antigen-binding fragment thereof, the antibody-drug conjugate or the pharmaceutical composition according to the present disclosure, optionally further comprising means for administering the antibody or antibody-drug conjugate to a subject and / or instructions for use.

[0132] Some embodiments of the present disclosure relate to the antibody or antigen-binding fragment thereof, the antibody-drug conjugate or the pharmaceutical composition according to the present disclosure for use in the manufacture of a medicament for treatment of a cancer, such as a cancer characterised by cells expressing MT1 -MMP, such as a cancer characterised by cancer cells, cancer-associated cells, such as P7363PC00 cancer-associated fibroblasts (CAFs), tumour cells and / or tumour-associated cells expressing MT1 -MMP.

[0133] Polynucleotides, vectors and host cells

[0134] Some embodiments of present disclosure relate to an isolated polynucleotide comprising a nucleotide sequence encoding an amino acid sequence comprising the antibody or antigen-binding fragment thereof according to the present disclosure.

[0135] Some embodiments of the present disclosure relate to an isolated polynucleotide comprising a nucleotide sequence encoding an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising or consisting of an amino acid sequence of SEQ ID NO: 1 or a sequence having at least 90% sequence identity thereto and an immunoglobulin heavy chain variable region comprising or consisting of an amino acid sequence of SEQ ID NO: 5 or a sequence having at least 90% sequence identity thereto, or ii. an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 or a sequence having at least 90% sequence identity thereto and an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27 or a sequence having at least 90% sequence identity thereto.

[0136] In some embodiments, the isolated polynucleotide comprising a nucleotide sequence encoding an antibody or antigen-binding fragment thereof comprising an immunoglobulin light chain variable region comprising or consisting of an amino acid sequence of SEQ ID NO: 1 and an immunoglobulin heavy chain variable region comprising or consisting of an amino acid sequence of SEQ ID NO: 5.

[0137] In some embodiments, the isolated polynucleotide comprising a nucleotide sequence encoding an antibody or antigen-binding fragment thereof comprising an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 and an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27. P7363PC00

[0138] Some embodiments of present disclosure relate to a vector, such as an expression vector, comprising the isolated polynucleotide according to the present disclosure.

[0139] In some embodiments of the present disclosure, the vector is a mammalian expression vector. In some embodiments, the vector is a plasmid vector, such as a plasmid vector selected from pD2610-v13 (ATUM), pSV and the pCMV series of plasmid vectors.

[0140] In some embodiments of the present disclosure, the vector is a viral vector, such as a viral vector selected from the group consisting of adenoviral vectors, lentiviral vectors, adeno-associated viral vectors, herpesviral vectors, vaccinia viral vectors, poxviral vectors, baculoviral vectors and oncolytic viral vectors.

[0141] Some embodiments of present disclosure relate to a host cell, such as a recombinant host cell, comprising the isolated polynucleotide and / or the vector according to the present disclosure.

[0142] In some embodiments of the present disclosure, the host cell is a recombinant host cell. In some embodiments of the present disclosure, the host cell comprising the polynucleotide and / or the vector as described herein is selected from the group consisting of CHO (Chinese hamster ovary) cells, COS (CV-1 (simian) in Origin, and carrying the SV40 genetic material) cells, HEK (Human embryonic kidney) cells and HeLa (Henrietta Lacks) cells.

[0143] Examples

[0144] Example 1. In vitro and in vivo efficacy of PNU-ADCs directed against MT1-MMP Methods

[0145] Preparation of anti-MT1 -MMP antibody-drug conjugates with PNU payload

[0146] Monoclonal antibodies (mAb) against MT1 -MMP were generated and produced using hybridoma technique after immunization of mice, following the established methods known in the art. The non-binding control mAb against HyHEL-10 was purchased (Sydlabs, Cat. No. PA007154.m1 , lot.220903-1 ). A second control mAb against trinitrophenol (TNP) has been described previously (Shulman et al., 1978).

[0147] In the case of mAb-2 against MT1 -MMP, the host mice that were immunized were deficient for MT1 -MMP, resulting in the creation of antibodies reactive to both the P7363PC00 murine and the human antigen. mAb-2 was obtained as part of the same immunization and cloning program as that leading to the published antibody, mAb-1 (Ingvarsen et al., 2008). The additional antibody against MT1 -MMP, 9E8:A7, has been published (Ingvarsen et al., 2013, J.Biol.Chem) and is referred to as 9E8 in Ingvarsen et al., 2013.

[0148] To prepare the antibodies for conjugation with PNU-159682, the N-glycosylation on the CH2 domain of each heavy chain was modified, followed by azide activation. For this procedure, the GlyClick Azide Activation kit (Genovis, Cat. No.: L1 -AZ1 -200) was utilized following the manufacturer’s instructions. The initial step of the azide activation process was the partial deglycosylation of the antibody, achieved by employing immobilized GlycINATOR®. After this, the exposed GIcNAc was modified with an azide utilizing Galactosyltransferase and UDP-GalNAz. After overnight incubation at 30°C, excess UDP-GalNAz was removed using a 2 ml Desalting Spin column. The azide- modified antibody was run on a reducing SDS-PAGE gel and visualized using Coomassie stain to confirm the hydrolysis of the N-glycan.

[0149] The azide-modified antibodies were then used for conjugation with the linker-toxin DBCO-PEG4-VC-PAB-DMEA-PNU-159682 (MCE, Cat. No.: HY-126691). Before conjugation, the linker-toxins were dissolved in a reaction buffer containing 20% N, N- dimethylformamide (DMF) (Sigma-Aldrich, Cat. No.: DX1730-6) and 80% propylene glycol (PG) (v / v) (Sigma-Aldrich, Cat. No.: P4347). The dissolved linker-toxin was added to the azide-modified antibody, dissolved in TBS, in a 15-fold molar excess. To ensure a final concentration of 5 and 20% (v / v) of DMF and PG, respectively, the conjugation reaction took place in a 1 :4 (v / v) ratio of linker-toxin (in 20 / 80 DMF / PG solution) to antibody (in 1X TBS, pH 7.4). After overnight incubation, the conjugated ADC was purified using 40K Zeba Spin Desalting Columns (Thermo Fisher, Cat. No.: A57759) and buffer exchanged to PBS, pH 7.4. The ADC’s successful conjugation was confirmed using Coomassie-stained reduced SDS-PAGE gel (data not shown) and the DAR was calculated with UV / vis spectroscopy using the QIAxpert system (QIAGEN). The drug-to antibody ratio (DAR) was calculated using the equation shown in Fig 9.

[0150] SDS-PAGE analysis of the conjugates

[0151] To confirm antibody conjugation, SDS-PAGE gels were used, specifically Bolt™ 4-12% Bis-Tris Plus gels (Invitrogen, Cat. No.: NW04125BOX). To prepare the samples for the cell lysates SDS-PAGE, 10 pg of total protein was mixed with 4x NuPAGE SDS sample buffer and PBS. For the conjugated antibodies, 1 pg of antibody was mixed with 4x P7363PC00

[0152] NuPAGE SDS sample buffer, DTT, and PBS. After mixing, the sample was incubated at 100°C for 5 minutes. The electrophoresis chamber was filled with 1 x MOPS (MOPS 20x, Invitrogen, Cat. No.: NP0001 -02) and the gel was loaded with the 12 pl of each sample as well as 5 pl of protein marker (Precision Plus Protein™ All Blue Prestained Protein Standards, Bio-Rad, Cat. No.: 161 -0373). The gel ran for 1 hour with a voltage of 200V.

[0153] For the Coomassie staining the SDS-PAGE gel was taken out of its cassette and transferred to a chamber containing Coomassie stain (0.1 % Coomassie Brilliant Blue R, 30 % EtOH, 10 % Acetic Acid). The gel was incubated for 30 minutes, followed by 4 x 20 minutes in destain (30% EtOH, 10% Acetic Acid). Afterward, the gel was incubated in 5% Acetic Acid overnight (Merck, Cat. No.: 1000631011 ). The gel was later washed in H2O and scanned using Odyssey® CLx Imaging System (LI-COR).

[0154] Cell lines

[0155] Different cell lines were used for the in vitro testing of ADCs comprising anti-MT 1 -MMP antibodies on a first instance: HT1080 (fibrosarcoma), MG-63 (osteosarcoma) and SaOS-2 (osteosarcoma). All cell lines were obtained from the American Type Culture Collection (ATCC). All cells were cultured in the appropriate medium with 10% fetal bovine serum and 1% penicillin / streptomycin and incubated in 37 °C and 5% C02.

[0156] Western blot analysis of different cell lines

[0157] Western blot analysis was employed to assess MT1 -MMP expression across a spectrum of sarcoma cell lines previously known to express the receptor. The cell lysates were prepared by washing the cells with PBS followed by incubation at 37°C for 5 minutes in PBS with 5 mM EDTA, to detach the cells. The cells were then harvested using PBS, counted, and pelleted by centrifugation at 180 x g for 5 minutes. The pellet was resuspended in 1 ml ice-cold lysis buffer per 107 cells (10 mM Tris, 140 mM NaCI, pH 7.4 + 1 % TX-100) with 1 :200 Protease Inhibitor Cocktail III (Millipore, Cat. No.: 535140) and transferred to an Eppendorf tube. This was followed by incubation for 15 minutes on ice and subsequent centrifugation at 20.000 x g for 30 minutes. Afterwards, the supernatants were transferred to a new Eppendorf tube and stored at -20°C.

[0158] The western blotting was performed with the iBIot system (Thermo Fisher). SDS-PAGE gels were taken out of their cassette and transferred to a PVDF iBIot blotting membrane (Thermo Fisher, Cat. No.: IB401001 ). Afterwards, the membrane was blocked for 1 hour at room temperature using PBS with 10% (w / w) skim milk (Sigma- P7363PC00

[0159] Aldrich, Cat. No.: 70166-500G) and 0.1% (v / v) Tween20 (Merck, Cat. No.: 8221840500). The membrane was then incubated with the primary antibody (1 pg / ml) (Abeam, Anti-MT1-MMP rabbit antibody, Cat. No.: ab51074, Lot.GR287144-3) in PBS + 0.1% Tween20 with 1% skim milk overnight at 4°C. After incubation with the primary antibody, the membrane was washed for 3 x 5 minutes PBS-T.

[0160] Subsequently, the membrane was incubated for 1 h at room temperature with a secondary antibody, conjugated to a fluorophore that emits at 800 nm (Goat anti-rabbit, LiCOR, Cat. No.: 925-32211 , Lot. D30322-11), in a final concentration of 1 .7 ng / ml in PBS-T with 1% skim milk. Finally, the membrane was washed 3 x 5 minutes in PBS-T and 2 x 2 minutes in miliQ H2O. The Odyssey® CLx Imaging System was used to scan the membrane.

[0161] In vitro cytotoxicity of ADCs - Cell viability assay

[0162] Cells were seeded out in 96-well plates with 3000-5000 cells per well in 100 ul complete culture medium. After 24 hours, the ADCs were added to the cells in a dilution series, specified in the figure section. The highest concentration of ADCs used was 5, 10, 15 or 35 pg / ml, depending on the ADC. As a control, in a single column on the plate the cells received medium without ADC. After three days of incubation at 37°C in a CO2 incubator, the cell viability was measured using the CellTiter 96® Aqueous One Solution Cell Proliferation Assay (MTS) (Promega, Cat. No.: G3582) (C. F. Nielsen et al., 2017). 15 ul of MTS reagent was added to each well, which makes the cells produce formazan depending on their viability. Following a 1 -hour incubation at 37°C in a CO2 incubator, the absorbance was recorded at 490 nm and 650 nm using a Spectra Max Plus plate reader.

[0163] T reatment of a subcutaneous MT 1 -MMP-fibrosarcoma xenograft tumor model in mice by intravenous injection of ADCs

[0164] All animal experiments were performed under legal approval from The Danish Veterinary and Food Administration, under license number 2020-15-0201 -00533 (L. H. Engelholm). In all experiments, animals received a standard of care according to the Danish animal welfare legislation. All mice were female CB17 severe combined immunodeficient (SCID) mice. All reagents and cell lines used in the in vivo study were previously tested and found negative for murine viruses, fungi, bacteria and mycoplasma. Animals received standard of care and were sacrificed upon loss of more P7363PC00 than 10% of body weight, visible stress illness or tumor grown over a volume of 1000 mm3.

[0165] Five-week-old mice received subcutaneous injections in the upper right flank with 1 .5x10A6 HT 1080 cells in 100 pL of PBS, leading to the development of tumors exhibiting rapid growth. Once tumors reached a volume of 80-150 mm3, mice were randomly assigned to one of three treatment groups: mAb-2-PNU (N=9), non-binding ADC control HyHEL-PNU (N=9), or PBS vehicle control (N=8). Tumor growth was monitored by palpation using electronic calipers, and volumes were calculated using the formula V=(LxW2) / 2, with L being the length of the tumor and W the width. Each mouse received three intravenous doses of 0.75 mg / kg ADC or control reagent via the tail vein, administered once every 7 days. The mice were euthanized when the tumor length reached above 12 mm (termination criteria), for which they received anesthesia (Zoletil) and were perfused by intracardial injection with 20 mL of PBS and relevant tissues were collected. Mice that exhibited tumor regression were subsequently examined weekly for 100 days.

[0166] Statistics

[0167] All graphs were conducted using GraphPad Prism 10. All data is given in (mean ± SD).

[0168] Results and conclusions

[0169] As mentioned in the Background section, the MT1 -MMP protein is upregulated in the tumor cells of specific cancers, including fibrosarcoma, osteosarcoma and several others, while in healthy individuals it presents a restricted expression. To establish tumor models with cell lines, we therefore first assessed the expression of MT1 -MMP in different sarcoma cell lines in western blot. The fibrosarcoma cell line HT1080, and osteosarcoma cell lines, SaOS-2 and MG63 showed a prominent expression of MT1- MMP, which made them good candidates for in vitro evaluation of the efficacy of ADCs targeting MT1 -MMP (data not shown).

[0170] Using the monoclonal antibody, mAb-2, obtained after immunization of a MT1 -MMP gene-deficient mice, we prepared an MT1 -MMP-directed ADC (mAb-2-PNU) as described in the Methods section above. This procedure results in a ratio of 2 molecules of toxin per mAb. When testing the resulting ADC in cytotoxicity assay in vitro, it demonstrated high specificity in targeting and killing MT1 -MMP-positive cancer cells, with the HT1080 cell line showing particularly high sensitivity (Fig. 2A, B). P7363PC00

[0171] Additionally, the osteosarcoma cell lines MG-63 and SaOS-2 were sensitive to mAb-2- PNU compared to the control ADC (Fig. 3)

[0172] The effect of the mAb-2 ADC was compared in HT1080 cells alongside ADCs comprising other anti-MT-MMP generated antibodies, 9E8:A7 and mAb-1 (S. Z. Ingvarsen et al., 2008). Notably, mAb-2-PNU demonstrated a cell-killing effect with an EC50 below 0.05 pg / ml (Fig. 4A). In contrast, although the ADCs based on the mAb-1 and 9E8:A7 clones also demonstrated cell-killing capacity, the EC50 with these ADCs was close to 0.5 pg / ml, while no such activity was observed at concentrations below 0.1 pg / ml (Fig. 4B and 4C). A comparison of the curves suggests that mAb-2-PNU is approximately ten times more potent than the ADCs comprising other anti-MT1 -MMP antibodies.

[0173] For the in vivo study, we used a subcutaneous xenograft tumor model with HT1080 cells in CD17 SCID mice. In this model, mAb-2-PNU was highly efficient at eradicating solid HT1080 tumors in vivo. After treatment with 3 intravenous doses of mAb-2-PNU, 7 out of 9 mice remained tumor-free for 90 days (Fig.5A) while all mice in the control groups were euthanized due to reaching the termination criteria. Thus, treatment with mAb-2-PNU resulted in a strongly increased survival (Fig.5B).

[0174] In conclusion, the data presented very strongly support the concept of the invention, that MT1-MMP is a promising target in ADC cancer therapy. The data presented show that an ADC comprising a specific antibody against MT1 -MMP, mAb-2-PNU, is highly effective for targeting MT1-MMP-expressing cells in vitro and in vivo.

[0175] Example 2. In vitro efficacy of mAb-2 ADC comprising an MMAE payload Methods

[0176] Preparation of anti-MT1 -MMP antibody-drug conjugates with MMAE payload Different toxins can be used in an ADC format targeting MT1 -MMP receptor. ADCs against various target proteins with MMAE as the cytotoxin are used in several clinically approved ADCs (Metrangolo & Engelholm (2024)). ADCs with MMAE as the cytotoxin and mAb-2 or HyHEL mAb as the antibody component were prepared using cysteine-linked conjugation. This antibody conjugation procedure was based on the protocol developed by Nielsen et al. (2017). The mAb-2 and HyHEL antibodies were buffer exchanged into a 50 mM NaBorate 50 mM NaCI buffer, pH 8.0 using 40K Zeba spin columns following the instructions of the supplier (Thermo Fisher, Cat. No.: A57759). To mildly reduce the antibodies, dithiothreitol (DTT) was added to a final concentration of 2.5 mM. After 30 minutes of incubation at 25°C on a shaking table, the P7363PC00 partially reduced antibodies were buffer exchanged to a PBS + 1 mM EDTA buffer, using 40K Zeba spin columns.

[0177] The conjugation was then performed with a linker-toxin MC-vc-PAB-MMAE (MedChemExpress, HY-15575). Linker-toxin was added to the partially reduced antibodies (in PBS) at a 5-times molar excess. The reaction mixture was incubated for 2 hours on a shaking table at 25°C. After incubation, Zeba spin columns 40k 0.5ml were used to remove excess unconjugated payload and purify the conjugated antibodies, following the instructions of the supplier.

[0178] The conjugation of the antibodies was visualized on a Coomassie-stained reduced SDS-PAGE gel by comparing the product with the unconjugated antibody (data not shown). The average DAR of the resulting ADCs was calculated using the equation showed in Fig.9, using the UV / vis absorbance measurements at 280 nm and 248 nm, respectively (Pozniak et al., 2021 ), measured using the QIAxpert system (QIAGEN). In vitro cell viability assays were performed as described in Example 1 and compared with mAb-2-PNU.

[0179] Results and conclusions

[0180] The assay performed on HT1080 cells reveals that mAb-2- MMAE can eradicate these cells with a notably higher efficiency than the non-binding control (Fig. 2C). When comparing the effect of the mAb-2 ADCs with the two different payloads studied, only a difference of approximately 5-fold is found (Fig.2D). This indicates that a mAb-2-based ADC with a clinically approved payload, MMAE, is also a relevant ADC candidate.

[0181] Example 3. In vitro efficacy of ADCs directed against MT1-MMP in other cancer cell lines

[0182] Methods

[0183] To support that mAb-2 ADCs are also useful for treatment of other cancer, the effect of mAb-2 ADCs was also tested on various non-sarcoma cancer cell lines. The preparation of the ADCs, with both PNU and MMAE were carried out as presented in Example 1 and 2, respectively. The in vitro viability assays were done as described in Example 1 .

[0184] Cell lines studied: U87-MG (malignant glioma), MDA-MB-231 (breast carcinoma), and EBC-1 (Lung squamous cell carcinoma) were all obtained from the American Type Culture Collection (ATCC). All cells were cultured in the appropriate medium with 10% fetal bovine serum and 1% penicillin / streptomycin and incubated in 37oC and 5% C02. P7363PC00

[0185] Results and conclusion

[0186] MT1 -MMP expression across various cancer cell lines was first confirmed by western blot analysis (data not shown). Subsequently, cell viability was tested in the presence of the ADCs mAb-2-PNU and mAb-2-MMAE. The breast carcinoma cell line MDA-MB- 231 demonstrated sensitivity to both mAb-2 ADCs (PNU and MMAE) compared to the negative control ADC (Fig. 6). Similarly, the lung carcinoma cell line EBC-1 showed sensitivity to both mAb-2 ADCs (PNU and MMAE) (Fig. 7), and the malignant glioma cell line U87-MG responded to mAb-2-PNU (Fig. 8).

[0187] These findings indicate that mAb-2 ADCs effectively target and kill cells across several different cancer cell types, highlighting the potential of this approach to treat various malignancies beyond sarcoma.

[0188] Example 4. ADC sensitivity assay, mAb-2-MMAE and -PNU, BxPC3 pancreatic adenocarcinoma cell line

[0189] Methods

[0190] The ADC preparation was carried out as presented in Example 1 and 2. The in vitro viability assays were done as described in Example 1 .

[0191] Cell line studied: BxPC3 (pancreatic adenocarcinoma) was obtained from the American Type Culture Collection (ATCC). Cells were cultured in the appropriate medium with 10% fetal bovine serum and 1% penicillin / streptomycin and incubated in 372C and 5% CO2.

[0192] Results

[0193] When evaluating the cell viability of BxPC3 in the presence of ADCs mAb-2-PNU and mAb-2-MMAE a clear therapeutic window between the mAb-2 and the respective negative control ADCs was observed (Fig. 10). These findings suggest the possibility of this approach to treat pancreatic adenocarcinoma.

[0194] Example 5. Treatment of a subcutaneous MDA-MB-231 -Triple Negative Breast Cancer xenograft tumor model in mice by intravenous injection of PNU-ADCs

[0195] Methods

[0196] The study was carried out as described in Example 1 with the following modifications: 2x106MDA-MB-231 cells in 100 pL of PBS were injected subcutaneously. mAb-2-PNU (N=9), non-binding ADC control HyHEL-PNU (N=8), and PBS vehicle control (N=8) P7363PC00

[0197] Results

[0198] A subcutaneous xenograft tumor model of triple-negative breast cancer (TNBC) was established using MDA-MB-231 cells implanted in CD17 SCID mice. In this model, treatment with mAb-2-PNU demonstrated high efficacy in controlling and slowing the growth of solid MDA-MB-231 tumors in vivo (Fig. 11 A) following the administration of three doses. Consequently, mAb-2-PNU treatment led to a significant improvement in survival (Fig. 11 B).

[0199] Example 6. Treatment of a subcutaneous EBC-1-Lung Squamous Cell Carcinoma xenograft tumor model in mice by intravenous injection of PNU and MMAE-ADCs Methods

[0200] The study was carried out as described in Example 1 with the following modifications: 1x106EBC-1 (Lung Squamous Cell Carcinoma) cells in 100 pL of PBS was injected subcutaneously. mAb-2-PNU (N=8), non-binding ADC control HyHEL-PNU (N=8) and PBS vehicle control (N=8). mAb-2-MMAE (N=8), non-binding ADC control HyHEL- MMAE (N=7), and PBS vehicle control (N=8). In the MMAE study, mice received three intravenous injections of 4 mg / kg ADC or control reagent following the same schedule as in Example 1 .

[0201] Results

[0202] In the PNU study, all mice treated with mAb-2-PNU exhibited tumor regression (Fig. 12A). However, three mice were euthanized early due to weight loss. PNU is a highly toxic payload and the ADC is dosed close to the maximum tolerated dose in mice. It can therefore be expected that a minor subset of the mice will experience side effects e.g. weight loss exceeding predetermined humane end points. In the three studies presented here, where mice were treated with mAb-2-PNU (See also Fig.5 and Fig.11 ), the majority of the mice tolerated the ADC. In addition, we also show that a mAb-2 ADC with another more clinically relevant payload, MMAE (Fig.13 and Fig.14), also shows potent anti-tumor effects, while being well tolerated by the mice. In the remaining animals, tumors gradually regrew after day 60. The mAb-2-PNU group showed a clear increase in survival time compared with the control groups (HyHEL- PNU and PBS) (Fig. 12B). P7363PC00

[0203] In the MMAE study, all mice treated with mAb-2-MMAE demonstrated complete tumor regression (Fig. 13A) with no signs of tumor regrowth, resulting in 100% survival throughout the study period (Fig. 13B).

[0204] In conclusion, these data strongly support that MT1 -MMP is a promising target for ADC-based cancer therapy, also in squamous cell lung carcinoma. These data also shows that this approach is efficacious in vivo with different cytotoxic payloads.

[0205] Example 7. Treatment of a subcutaneous U-87 MG Glioblastoma xenograft tumor model in mice by intravenous injection of MMAE-ADCs

[0206] Methods

[0207] The study was carried out as described in Example 1 with the following modifications: 2x106U-87 MG (Glioblastoma) cells in 100 pL of PBS were injected subcutaneously. Mice were treated with 4 mg / kg MMAE-conjugated ADC in the following three treatment groups: mAb-2-MMAE (N=4), non-binding isotype control (IC) ADC IC-MMAE (N=4), and PBS vehicle control (N=4).

[0208] Results

[0209] As shown in Fig. 14A, mAb-2-MMAE treatment resulted in improved tumor growth control compared to both the PBS and IC-MMAE control groups. Tumor progression was significantly delayed in mice receiving mAb-2-MMAE. Correspondingly, the survival analysis (Fig. 14B) demonstrated a substantial extension of survival for the mice treated with mAb-2-MMAE.

[0210] Example 8. ADC sensitivity assay, mAb-2-MMAE and DX2400-MMAE, HT1080 fibrosarcoma cell line and EBC-1 lung squamous cell carcinoma cell line

[0211] Methods

[0212] The ADC preparation was carried out as presented in Example 2, with mAb-2 and DX2400 as the antibody component and MMAE as the cytotoxin.

[0213] The DX2400 antibody was obtained from MedChem Express, cat.no. HY-P991316.

[0214] The in vitro cytotoxicity assays were done as described in Example 1 using cell lines HT1080 (fibrosarcoma) and EBC-1 (lung squamous cell carcinoma) and adding a maximum concentration of 10 pg / ml ADC in a three-fold serial dilution series. P7363PC00

[0215] Results

[0216] The two antibodies mAb-2 and DX2400 were both conjugated with the cytotoxin MMAE using identical conditions, resulting in highly comparable ADCs with indistinguishable drug-to-antibody ratios (Table 1 ).

[0217] Table 1 Drug-to-antibody ratio (DAR) of mAb-2-MMAE and DX2400-MMAE.

[0218] Table 1 ; The DAR of the two ADCs were calculated based on uv / vis spectroscopy, measuring absorbance at 280 nm and 248 nm and using the equation in Fig.9. The analysis revealed the DAR to be indistinguishable (within the experimental uncertainty) for the two ADCs.

[0219] When comparing their ability to kill cancer cell lines in vitro, the mAb-2-MMAE was found to have superior performance over DX2400-MMAE. For both fibrosarcoma cell line, HT 1080, and lung squamous cell carcinoma cell line, EBC-1 , 50% of the cells were eradicated at a three-fold lower concentration of mAb-2-MMAE compared to DX2400-MMAE (Fig. 15).

[0220] Sequence overview

[0221] All CDR regions in the below table have been determined using the IMGT-annotation tool IMGT / V-QUEST (Brochet et al., 2008; Giudicelli et al., 2011 ). P7363PC00 P7363PC00 P7363PC00 P7363PC00 P7363PC00

[0222] P7363PC00

[0223] Items

[0224] 1 . An antibody or antigen-binding fragment thereof which binds to Membrane type 1 -matrix metalloproteinase (MT1 -MMP), wherein said antibody is: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 2, as CDR2 an amino acid sequence of SEQ ID NO: 3, and as CDR3 and amino acid sequence of SEQ ID NO: 4, and ii. an immunoglobulin heavy chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 6, as CDR2 an amino acid sequence of SEQ ID NO: 7, and as CDR3 and amino acid sequence of SEQ ID NO: 8, or b. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 1 or a sequence having at least 90% sequence identity thereto, and ii. an immunoglobulin heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 5 or a sequence having at least 90% sequence identity thereto, or c. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 or a sequence having at least 90% sequence identity thereto, and ii. an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27 or a sequence having at least 90% sequence identity thereto, or d. a humanized version of the antibody or antigen-binding fragment thereof of any one of a, b or c, or e. a chimeric version of the antibody or antigen-binding fragment thereof of any one of a, b or c.

[0225] 2. The antibody or antigen-binding fragment thereof according to item 1 , wherein any sequence variance is outside the complementarity-determining regions. P7363PC00

[0226] 3. The antibody or antigen-binding fragment thereof according to any of the preceding items, wherein the antibody or antigen-binding fragment thereof comprises: a. an immunoglobulin light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 1 , and b. an immunoglobulin heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 5.

[0227] 4. The antibody or antigen-binding fragment thereof according to any of the preceding items, wherein the antibody or antigen-binding fragment thereof comprises: a. an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26, and b. an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27.

[0228] 5. The antibody or antigen-binding fragment thereof according to any of the preceding items, wherein the antibody is selected from a mouse antibody, a chimeric antibody, a human antibody, a humanised antibody, a humanised antigen-binding fragment, a Fab fragment, a Fab’ fragment, an F(ab’)2 fragment, an Fv, a single chain antibody (SCA) such as an scFv, the variable portion of the heavy and / or light chains thereof, and a Fab miniantibody.

[0229] 6. The antibody or antigen-binding fragment thereof according to any of the preceding items, wherein the antibody is a monoclonal antibody.

[0230] 7. The antibody or antigen-binding fragment thereof according to any of the preceding items, wherein the antibody is a humanised or fully human monoclonal antibody or antigen-binding fragment thereof.

[0231] 8. The antibody or antigen-binding fragment thereof according to any of the preceding items, wherein the antibody is a recombinant antibody. P7363PC00

[0232] 9. The antibody or antigen-binding fragment thereof according to any one of the preceding items, wherein said antibody is an immunoglobulin class antibody such as IgG, IgM, IgD, IgE, IgA, and any subclass thereof, such as lgG1 , lgG2, lgG3 or lgG4.

[0233] 10. An antibody-drug conjugate (ADC) comprising: a. the antibody or antigen-binding fragment thereof as defined in any one of the preceding items, b. an active agent, and c. optionally a linker which links a) to b).

[0234] 11 . The antibody-drug conjugate according to item 10, wherein the active agent is selected from a therapeutic agent, a cytotoxic agent, a radioisotope, and a detectable label.

[0235] 12. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is a cytotoxic agent.

[0236] 13. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is a therapeutic agent, such as a therapeutic agent selected from the group consisting of anti-microtubule / anti-mitotic agents, DNA crosslinking agents, DNA alkylating agents, DNA strand scission agents, anthracyclines, antimetabolites, histone deacetylase inhibitors, kinase inhibitors, metabolism inhibitors, peptide antibiotics, immune checkpoint inhibitors, platinum-based antineoplastics, topoisomerase inhibitors, DNA or RNA polymerase inhibitors, nucleotide based agents, and cytotoxic antibiotics.

[0237] 14. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is an anti-mitotic agent, such as selected from the group consisting of derivatives of auristatin or dolastatin such as monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF) and more, a taxane such as Paclitaxel or Docetaxel and more, a vinca alkaloid such as Vinblastine, Vincristine, Vindesine or Vinorelbine and more, a mayatansinoid, Colchicine, and Podophyllotoxin. P7363PC00

[0238] 15. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is monomethyl auristatin E (MMAE).

[0239] 16. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is a DNA-crosslinking agent, such as a DNA crosslinking agent selected from cisplatin or a derivative of cisplatin such as carboplatin or oxaliplatin, mitomycin C (MMC), pyrrolobenzodiazepine, and dimeric pyrrolobenzodiazepine derivatives such as SGD-1882.

[0240] 17. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is a DNA alkylating agent, such as a DNA alkylating agent selected from nitrogen mustards such as tris(2-chloroethyl)amine, pyridinobenzodiazepines or a pyridinobenzodiazepine derivative, indolinobenzodiazepine dimers, and Duocarmycin SA.

[0241] 18. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is a DNA strand scission agent, such as a DNA strand scission agent selected from calicheamicin and hamiltrone.

[0242] 19. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is an anthracycline, such as an anthracycline selected from Daunorubicin, doxorubicin, epirubicin, idarubicin, and PNU-159682.

[0243] 20. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is PNU-159682.

[0244] 21 . The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is an antimetabolite, such as an antimetabolite selected from folic acid antagonists such as methotrexate, purine antimetabolites such as 6-mercaptopurine or 6-thioguanine or fludarabine phosphate or pentostatin or cladribine, and pyrimidine antimetabolites such as 5-fluorouracil or 5-fluorodeoxyuridine or cytarabine or gemcitabine.

[0245] 22. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is a histone deacetylase inhibitor, such as a histone P7363PC00 deacetylase inhibitor selected from trichostatin A, vorinostat, belinostat, panabiostat, givinostat, resminostat, abexinostat, quisinostat, rocilinostat, practinostat, CHR-3996, valproic acid, butyric acid, phenylbutyric acid, entinostat, tacedinaline, 4SC202, mocetinostat, romidepsin, nicotinamide, sirtinol, cambinol, and EX-527.

[0246] 23. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is a kinase inhibitor, such as a kinase inhibitor selected from genistein, lavendustin C, PP1 -AG1872, PP2-AG1879, SU6656, CGP77675, PD166285, imatinib, erlotinib, gefitinib, lavendustin A, cetuximab, UCS15A, herbimycin A, and radicicol.

[0247] 24. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is a metabolism inhibitor, such as an NAM PT inhibitor selected from APO866, GMX-1777, GMX-1778 ATG-019, and OT-82.

[0248] 25. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is an immune checkpoint inhibitor, such as a PD-1 inhibitor selected from Pembrolizumab, Nivolumab, Cemiplimab, JTX-4014, Spartalizumab, Camrelizumab, Sintilimab, Tislelizumab, Toripalimab, Dostarlimab, AMP-224 and AMP-514; or a PD-L1 inhibitor selected from Atezolizumab, Avelumab, Durvalumab, KN035, CK-301 , AUNP12, CA-170 and BMS-986189.

[0249] 26. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is a platinum-based antineoplastic, such as a platinum-based antineoplastic selected from lipoplatin, cisplatin, carboplatin, oxaliplatin, nedaplatin, picoplatin, phenanthriplatin, satraplatin, and triplatin tetranitrate.

[0250] 27. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is a topoisomerase inhibitor, such as a topoisomerase inhibitor selected from camptothecin or derivatives thereof such as topotecan, belotecan, lurtotecan, irinotecan, SN-38, exatecan, and Dxd. P7363PC00

[0251] 28. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent is a DNA- or RNA-polymerase inhibitor, such as a polymerase inhibitor selected from amanitin or alpha-amanitin or derivatives thereof, actinomycin D, and aphidicolin.

[0252] 29. The antibody-drug conjugate according to any one of the preceding items, wherein the active agent comprises a radioisotope selected from60Co,89Sr,90Y, 99mTc,1311,137Cs,153Sm, and223Rd.

[0253] 30. The antibody-drug conjugate according to any one of the preceding items, wherein the drug-to-antibody ratio (DAR) is between 1 and 10, such as between 2 and 8, such as between 2 and 6, such as between 2 and 4.

[0254] 31 . The antibody-drug conjugate according to any one of the preceding items, wherein the drug-to-antibody ratio (DAR) is 2 or 4.

[0255] 32. The antibody-drug conjugate according to any one of the preceding items, wherein the antibody-drug conjugate comprises a linker selected from a cleavable and a non-cleavable linker.

[0256] 33. The antibody-drug conjugate according to any one of the preceding items, wherein the antibody-drug conjugate comprises a peptide linker.

[0257] 34. The antibody-drug conjugate according to any one of the preceding items, wherein the linker comprises a dipeptide.

[0258] 35. The antibody-drug conjugate according to any one of the preceding items, wherein the linker comprises valine-citrulline (VC) and / or valine-alanine (VA).

[0259] 36. The antibody-drug conjugate according to any one of the preceding items, wherein the linker comprises p-aminobenzoic acid (PAB), p- aminobenzylcarbamate (PABC), p-aminobenzoyloxycabonyl, or polyethylenglycol (PEG). P7363PC00

[0260] 37. The antibody-drug conjugate according to any one of the preceding items, wherein the linker comprises p-aminobenzoic acid (PAB).

[0261] 38. The antibody-drug conjugate according to any one of the preceding items, wherein the linker comprises a dibenzocyclooctyne (DBCO) including amine- functionalized dibenzocyclooctynes (DBCO), optionally modified with a PEG, such as PEG4, maleimide, N-hydroxysuccinimide, modified or unmodified protein-bound carbohydrate, peptide sequences that are required for enzymatic reactions, azides or alkynes or being derived from these by reaction with the antibody or a chemically or enzymatically generated derivative thereof.

[0262] 39. The antibody-drug conjugate according to any one of the preceding items, wherein the linker comprises dibenzocyclooctyne (DBCO), such as an amine- functionalized dibenzocyclooctyne (DBCO), and PEG4.

[0263] 40. The antibody-drug conjugate according to any one of the preceding items, wherein the linker comprises maleimidocaproyl (MC), optionally modified with a PEG, such as PEG4, maleimide, N-hydroxysuccinimide, modified or unmodified protein-bound carbohydrate, peptide sequences that are required for enzymatic reactions, azides or alkynes or being derived from these by reaction with the antibody or a chemically or enzymatically generated derivative thereof.

[0264] 41 . The antibody-drug conjugate according to any one of the preceding items, wherein the linker comprises maleimidocaproyl (MC).

[0265] 42. The antibody-drug conjugate according to any one of the preceding items, wherein the antibody-drug conjugate comprises or consists of: a. the antibody or antigen-binding fragment thereof as defined in any one of items 1 to 9, b. valine-citrulline (VC), c. dibenzocyclooctyne-amine (DBCO), optionally N-functionalized with PEG4, d. PAB, and e. PNU-159682. P7363PC00

[0266] 43. The antibody-drug conjugate according to any one of the preceding items, wherein the antibody-drug conjugate comprises or consists of: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 2, as CDR2 an amino acid sequence of SEQ ID NO: 3, and as CDR3 and amino acid sequence of SEQ ID NO: 4, and ii. an immunoglobulin heavy chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 6, as CDR2 an amino acid sequence of SEQ ID NO: 7, and as CDR3 and amino acid sequence of SEQ ID NO: 8, b. valine-citrulline (VC), c. dibenzocyclooctyne-amine (DBCO), optionally N-functionalized with PEG4, d. PAB, and e. PNU-159682.

[0267] 44. The antibody-drug conjugate according to any one of the preceding items, wherein the antibody-drug conjugate comprises or consists of: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 1 and an immunoglobulin heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 5, or ii. an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 and an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27, b. valine-citrulline (VC), c. dibenzocyclooctyne-amine (DBCO), optionally N-functionalized with PEG4, d. PAB, and e. PNU-159682. P7363PC00

[0268] 45. The antibody-drug conjugate according to any one of the preceding items, wherein the antibody-drug conjugate comprises or consists of: a. the antibody or antigen-binding fragment thereof as defined in any one of items 1 to 9, b. maleimidocaproyl (MC), c. valine-citrulline (VC), d. PAB, and e. monomethyl auristatin E (MMAE).

[0269] 46. The antibody-drug conjugate according to any one of the preceding items, wherein the antibody-drug conjugate comprises or consists of: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 2, as CDR2 an amino acid sequence of SEQ ID NO: 3, and as CDR3 and amino acid sequence of SEQ ID NO: 4, and ii. an immunoglobulin heavy chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 6, as CDR2 an amino acid sequence of SEQ ID NO: 7, and as CDR3 and amino acid sequence of SEQ ID NO: 8, b. maleimidocaproyl (MC), c. valine-citrulline (VC), d. PAB, and e. monomethyl auristatin E (MMAE).

[0270] 47. The antibody-drug conjugate according to any one of the preceding items, wherein the antibody-drug conjugate comprises or consists of: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 1 and an immunoglobulin heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 5, or P7363PC00 ii. an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 and an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 275, b. maleimidocaproyl (MC), c. valine-citrulline (VC), d. PAB, and e. monomethyl auristatin E (MMAE).

[0271] 48. The antibody-drug conjugate according to any one of the preceding items, wherein the antibody-drug conjugate comprises or consists of: a. the antibody or antigen-binding fragment thereof as defined in any one of items 1 to 9, and b. DBCO-PEG4-VC-PAB-DMAE-PNU159682.

[0272] 49. The antibody-drug conjugate according to any one of the preceding items, wherein the antibody-drug conjugate comprises or consists of: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 2, as CDR2 an amino acid sequence of SEQ ID NO: 3, and as CDR3 and amino acid sequence of SEQ ID NO: 4, and ii. an immunoglobulin heavy chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 6, as CDR2 an amino acid sequence of SEQ ID NO: 7, and as CDR3 and amino acid sequence of SEQ ID NO: 8, and b. DBCO-PEG4-VC-PAB-DMAE-PNU159682.

[0273] 50. The antibody-drug conjugate according to any one of the preceding items, wherein the antibody-drug conjugate comprises or consists of: a. an antibody or antigen-binding fragment thereof comprising: P7363PC00 i. an immunoglobulin light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 1 and an immunoglobulin heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 5, or ii. an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 and an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27, and b. DBCO-PEG4-VC-PAB-DMAE-PNU159682.

[0274] 51 . The antibody-drug conjugate according to any one of the preceding items, wherein the antibody-drug conjugate comprises or consists of the following structure or a regioisomer thereof: wherein the wiggled line denotes any chemical bonding to the antibody.

[0275] 52. The antibody-drug conjugate according to any one of the preceding items, wherein the antibody-drug conjugate comprises or consists of: a. the antibody or antigen-binding fragment thereof as defined in any one of items 1 to 9, and b. MC-vc-PAB-MMAE. P7363PC00

[0276] 53. The antibody-drug conjugate according to any one of the preceding items, wherein the antibody-drug conjugate comprises or consists of: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 2, as CDR2 an amino acid sequence of SEQ ID NO: 3, and as CDR3 and amino acid sequence of SEQ ID NO: 4, and ii. an immunoglobulin heavy chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 6, as CDR2 an amino acid sequence of SEQ ID NO: 7, and as CDR3 and amino acid sequence of SEQ ID NO: 8, and b. MC-vc-PAB-MMAE.

[0277] 54. The antibody-drug conjugate according to any one of the preceding items, wherein the antibody-drug conjugate comprises or consists of: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 1 and an immunoglobulin heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 5, or ii. an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 and an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27, and b. MC-vc-PAB-MMAE.

[0278] 55. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof according to any one of items 1 to 9 or the antibody-drug conjugate according to any one of items 10 to 54 and a pharmaceutically acceptable buffer, diluent, carrier, adjuvant or excipient. P7363PC00

[0279] 56. The antibody or antigen-binding fragment thereof according to any one of items 1 to 9, the antibody-drug conjugate according to any one of items 10 to 54 or the pharmaceutical composition according to item 55 for use as a medicament.

[0280] 57. The antibody or antigen-binding fragment thereof according to any one of items 1 to 9, the antibody-drug conjugate according to any one of items 10 to 54 or the pharmaceutical composition according to item 55 for use in the treatment of a cancer, such as a cancer characterised by cells expressing MT1 -MMP, such as a cancer characterised by cancer cells, cancer-associated cells, such as cancer-associated fibroblasts (CAFs), tumour cells and / or tumour-associated cells expressing MT1 -MMP.

[0281] 58. The antibody or antigen-binding fragment thereof, antibody-drug conjugate or composition for use according to item 57, wherein the cancer is fibrosarcoma, osteosarcoma, malignant glioma, lung squamous cell carcinoma or breast carcinoma.

[0282] 59. The antibody or antigen-binding fragment thereof, antibody-drug conjugate or composition for use according to item 57, wherein the cancer is pancreatic cancer, breast cancer, colon cancer, gastric cancer, esophageal cancer, hepatocellular cancer, prostate cancer, bladder cancer, bone cancer, lung cancer, mesothelioma or squamous cell carcinomas.

[0283] 60. The antibody or antigen-binding fragment thereof, antibody-drug conjugate or composition for use according to item 57, wherein the cancer is pancreatic adenocarcinoma (PAAD) such as pancreatic ductal adenocarcinoma (PDAC), bladder urothelial carcinoma (BLCA), breast invasive carcinoma (BRCA), cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC), cholangiocarcinoma (CHOL), colon adenocarcinoma (COAD), esophageal carcinoma (ESCA), glioblastoma multiforme (GBM), head and neck squamous cell carcinoma (HNSCC), acute myeloid leukemia (LAML), ovarian serous cystadenocarcinoma (OV), rectum adenocarcinoma (READ), sarcoma (SARC), skin cutaneous melanoma (SKCM) or stomach adenocarcinoma (STAD). P7363PC00

[0284] 61. The antibody or antigen-binding fragment thereof, antibody-drug conjugate or composition for use according to item 57, wherein the cancer is breast cancer, such triple-negative breast cancer (TNBC), basal-like breast cancer (BLBC), HER-2 breast cancer, Luminal A type breast cancer or Luminal B type breast cancer.

[0285] 62. The antibody or antigen-binding fragment thereof, antibody-drug conjugate or composition for use according to any one of items 57 to 61 , wherein the cancer is metastatic cancer.

[0286] 63. The antibody or antigen-binding fragment thereof, antibody-drug conjugate or composition for use according to any one of items 57 to 62, wherein administration is parenteral.

[0287] 64. The antibody or antigen-binding fragment thereof, antibody-drug conjugate or composition for use according to any one of items 57 to 63, wherein administration is intravenously, intracerebroventricularly, intraarticularly, intraarterially, intraperitoneally, intrathecally, intraventricularly, intrasternally, intracranially, intramuscularly or subcutaneously or by infusion techniques.

[0288] 65. The antibody or antigen-binding fragment thereof, antibody-drug conjugate or composition for use according to any one of items 57 to 63, wherein administration is intravenously.

[0289] 66. The antibody or antigen-binding fragment thereof, antibody-drug conjugate or composition for use according to any one of items 57 to 65, wherein the antibody or antigen-binding fragment thereof, antibody-drug conjugate or composition is administered in combination with one or more further agents, such as one or more further therapeutic agents.

[0290] 67. The antibody or antigen-binding fragment thereof, antibody-drug conjugate or composition for use according to any one of items 57 to 66, wherein the cells expressing MT1 -MMP display MT1-MMP overexpression, optionally wherein the cells expressing MT1-MMP are cancer cells, cancer-associated cells, such P7363PC00 as cancer-associated fibroblasts (CAFs), tumour cells and / or tumour-associated cells.

[0291] 68. The antibody or antigen-binding fragment thereof, antibody-drug conjugate or composition for use according to any one of items 57 to 67, wherein the antibody, antibody-drug conjugate or composition induces cell death and / or inhibits the growth and / or proliferation of the cells expressing MT1-MMP, such as cancer cells, cancer-associated cells, such as cancer-associated fibroblasts (CAFs), tumour cells and / or tumour-associated cells expressing MT1 -MMP.

[0292] 69. The antibody or antigen-binding fragment thereof, antibody-drug conjugate or composition for use according to any one of items 57 to 68, wherein the treatment is ameliorative or curative.

[0293] 70. The antibody or antigen-binding fragment thereof according to any one of items 1 to 9, the antibody-drug conjugate according to any one of items 10 to 54 or the pharmaceutical composition according to item 55 for use in a method for inhibiting tumour progression in a subject.

[0294] 71 . A kit comprising the antibody or antigen-binding fragment thereof according to any one of items 1 to 9, the antibody-drug conjugate according to any one of items 10 to 54 or the pharmaceutical composition according to item 55, optionally further comprising means for administering the antibody or antibodydrug conjugate to a subject and / or instructions for use.

[0295] 72. The antibody or antigen-binding fragment thereof according to any one of items 1 to 9, the antibody-drug conjugate according to any one of items 10 to 54 or the pharmaceutical composition according to item 55 for use in the manufacture of a medicament for treatment of a cancer, such as a cancer characterised by cells expressing MT1 -MMP, such as a cancer characterised by cancer cells, cancer-associated cells, such as cancer-associated fibroblasts (CAFs), tumour cells and / or tumour-associated cells expressing MT1 -MMP. P7363PC00

[0296] 73. An isolated polynucleotide comprising a nucleotide sequence encoding an amino acid sequence comprising the antibody or antigen-binding fragment thereof according to any one of items 1 to 9. 74. A vector, such as an expression vector, comprising the isolated polynucleotide according to item 73.

[0297] 75. A host cell, such as a recombinant host cell, comprising the isolated polynucleotide according to item 73 and / or the vector according to item 74.

[0298] P7363PC00

[0299] References

[0300] Brochet, X., Lefranc, M.-P. & Giudicelli, V. (2008). IMGT / V-QUEST: the highly customized and integrated system for IG and TR standardized V-J and V-D-J sequence analysis. Nucleic Acids Research, 36, W503-508 PMID: 18503082. DOI: 10.1093 / nar / gkn316

[0301] Chen, Y. (2014). Drug-to-Antibody Ratio (DAR) by UV / Vis Spectroscopy. In Laurent Ducry (ed.), Antibody-Drug Conjugates, Methods in Molecular Biology, vol. 1045 (Vol. 1045, pp. 267-273). https: / / doi.org / 10.1007 / 978-1-62703-541 -5

[0302] Chun, T. H., Sabeh, F., Ota, I., Murphy, H., McDonagh, K. T., Holmbeck, K., Birkedal- Hansen, H., Allen, E. D., & Weiss, S. J. (2004). MT1 -MMP-dependent neovessel formation within the confines of the three-dimensional extracellular matrix. Journal of Cell Biology, 167(4), 757-767. https: / / doi.org / 10.1083 / jcb.200405001

[0303] Dumontet, C., Reichert, J. M., Senter, P. D., Lambert, J. M., & Beck, A. (2023). Antibodydrug conjugates come of age in oncology. Nature Reviews Drug Discovery, 22(8), 641 — 661 . https: / / doi.Org / 10.1038 / S41573-023-00709-2

[0304] Giudicelli, V., Brochet, X., Lefranc, M.-P. (2011). Cold Spring Harbor Protocols, Jun 1 , (6). pii: pdb.prot5633. doi: 10.1101 / pdb.prot5633.

[0305] Gonzalez-Molina, J., Gramolelli, S., Liao, Z., Carlson, J. W., Ojala, P. M., & Lehti, K. (2019). MMP14 in Sarcoma: A regulator of tumor microenvironment communication in connective tissues. Cells, 8(9). https: / / doi.org / 10.3390 / cells8090991

[0306] Ingvarsen, S., Madsen, D. H., Hillig, T., Lund, L. R., Holmbeck, K., Behrendt, N., & Engelholm, L. H. (2008). Dimerization of endogenous MT1 -MMP is a regulatory step in the activation of the 72-kDa gelatinase MMP-2 on fibroblasts and fibrosarcoma cells. Biological Chemistry, 389(7), 943-953. https: / / 7

[0307] Ingvarsen, S. et al. (2013) «Targeting a single function of the multifunctional matrix metalloprotease MT1 -MMP Impact on Lymphangiogenesis», Journal of Biological Chemistry, 288(15), pp. 10195-10204. doi: 10.1074 / jbc.M112.447169.

[0308] Purcell et al 2018. LRRC15 Is a Novel Mesenchymal Protein and Stromal Target for Antibody-Drug Conjugates. Cancer Res (2018) 78 (14): 4059-4072. https: / / doi.Org / 10.1158 / 0008-5472.CAN-18-0327

[0309] Metrangolo, V., & Engelholm, L. H. (2024). Antibody - Drug Conjugates: The Dynamic Evolution from Conventional to Next-Generation Constructs. Cancers. 16(2). https: / / d0i.0rg / l 0.3390 / cancers16020447.

[0310] Pozniak, M., Por^bska, N., Krzyscik, M. A., Sokolowska-W^dzina, A., Jastrz^bski, K., Sochacka, M., Szymczyk, J., Zakrzewska, M., Otlewski, J., & Opalihski, L. (2021 ). The P7363PC00 cytotoxic conjugate of highly internalizing tetravalent antibody for targeting FGFR1- overproducing cancer cells. Molecular Medicine, 27(1 ). https: / / doi.org / 10.1186 / s10020- 021 -00306-2

[0311] Shulman, M., Wilde, C.D., Kohler, G. (1978). A better cell line for making hybridomas secreting specific antibodies. Nature, 276(5685), 270-272. https: / / doi.Org / 10.1038 / 276270a0

[0312] Turunen, S. P., Tatti-Bugaeva, O., & Lehti, K. (2017). Membrane-type matrix metalloproteases as diverse effectors of cancer progression. Biochimica et Biophysica Acta - Molecular Cell Research, 1864( ), 1974-1988. https: / / d0i.0rg / l 0.1016 / j.bbamcr.2O17.04.002

Claims

62P7363PC00Claims1 . An antibody or antigen-binding fragment thereof which binds to Membrane type 1 -matrix metalloproteinase (MT1 -MMP), wherein said antibody is: a. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 2, as CDR2 an amino acid sequence of SEQ ID NO: 3, and as CDR3 and amino acid sequence of SEQ ID NO: 4, and ii. an immunoglobulin heavy chain variable region comprising as complementarity-determining region 1 (CDR1 ) an amino acid sequence of SEQ ID NO: 6, as CDR2 an amino acid sequence of SEQ ID NO: 7, and as CDR3 and amino acid sequence of SEQ ID NO: 8, or b. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 1 or a sequence having at least 90% sequence identity thereto, and ii. an immunoglobulin heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 5 or a sequence having at least 90% sequence identity thereto, or c. an antibody or antigen-binding fragment thereof comprising: i. an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 or a sequence having at least 90% sequence identity thereto, and ii. an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27 or a sequence having at least 90% sequence identity thereto, or d. a humanized version of the antibody or antigen-binding fragment thereof of any one of a, b or c, or e. a chimeric version of the antibody or antigen-binding fragment thereof of any one of a, b or c, wherein any sequence variance is outside the complementaritydetermining regions.63P7363PC002. The antibody or antigen-binding fragment thereof according to claim 1 , wherein the antibody or antigen-binding fragment thereof comprises: a. an immunoglobulin light chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 1 and an immunoglobulin heavy chain variable region comprising or consisting of the amino acid sequence of SEQ ID NO: 5, or b. an immunoglobulin light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 26 and an immunoglobulin heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 27.

3. An antibody-drug conjugate (ADC) comprising: a. the antibody or antigen-binding fragment thereof as defined in any one of claims 1 or 2, b. an active agent, and c. optionally a linker which links a) to b).

4. The antibody-drug conjugate according to claim 3, wherein the active agent is selected from a therapeutic agent, a cytotoxic agent, a radioisotope, and a detectable label.

5. The antibody-drug conjugate according to any one of claims 3 to 4, wherein the active agent is a cytotoxic agent.

6. The antibody-drug conjugate according to any one of claims 3 to 5, wherein the active agent is an anthracycline, such as an anthracycline selected from Daunorubicin, doxorubicin, epirubicin, idarubicin, and PNU-159682.

7. The antibody-drug conjugate according to any one of claims 3 to 6, wherein the active agent is PNU-159682.

8. The antibody-drug conjugate according to any one of claims 3 to 7, wherein the antibody-drug conjugate comprises or consists of: a. the antibody or antigen-binding fragment thereof as defined in any one of claims 1 or 2,64P7363PC00 b. valine-citrulline (VC), c. dibenzocyclooctyne-amine (DBCO), optionally N-functionalized with PEG4, d. PAB, and e. PNU-159682.

9. The antibody-drug conjugate according to any one of claims 3 to 8, wherein the antibody-drug conjugate comprises or consists of the following structure or a regioisomer thereof:wherein the wiggled line denotes any chemical bonding to the antibody.

10. The antibody-drug conjugate according to any one of claims 3 to 5, wherein the active agent is an anti-mitotic agent, such as selected from the group consisting of derivatives of auristatin or dolastatin such as monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF) and more, a taxane such as Paclitaxel or Docetaxel and more, a vinca alkaloid such as Vinblastine, Vincristine, Vindesine or Vinorelbine and more, a mayatansinoid, Colchicine, and Podophyllotoxin.11 . The antibody-drug conjugate according to claim 10, wherein the active agent is monomethyl auristatin E (MMAE).65P7363PC0012. The antibody-drug conjugate according to any one of claims 3 to 5 or 10 to 11 , wherein the antibody-drug conjugate comprises or consists of: a. the antibody or antigen-binding fragment thereof as defined in any one of claims 1 or 2, b. maleimidocaproyl (MC), c. valine-citrulline (VC), d. PAB, and e. monomethyl auristatin E (MMAE).

13. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof according to any one of claims 1 to 2 or the antibody-drug conjugate according to any one of claims 3 to 12 and a pharmaceutically acceptable buffer, diluent, carrier, adjuvant or excipient.

14. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 2, the antibody-drug conjugate according to any one of claims 3 to 12 or the pharmaceutical compositions according to claim 13 for use as a medicament.

15. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 2, the antibody-drug conjugate according to any one of claims 3 to 12 or the pharmaceutical compositions according to claim 13 for use in the treatment of a cancer, such as a cancer characterised by cells expressing MT1 - MMP, such as a cancer characterised by cancer cells, cancer-associated cells, such as cancer-associated fibroblasts (CAFs), tumour cells and / or tumour- associated cells expressing MT1 -MMP.

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