阅读说明：本技术 纤溶酶可切割的抗不溶性纤维蛋白的抗体与药物的偶联物 (Conjugates of plasmin-cleavable anti-insoluble fibrin antibody and drug ) 是由 松村保广 真锅史乃 渊上弥史 于 2018-04-27 设计创作，主要内容包括：本发明涉及抗体药物偶联物(ADC)以及包含该偶联物的用于治疗癌症的组合物。本发明提供在针对不溶性纤维蛋白的特异性抗体与药物的ADC中,连接抗体与药物的接头具有纤溶酶切割序列的ADC以及包含该ADC的用于治疗癌症的药物组合物。(The present invention relates to Antibody Drug Conjugates (ADCs) and compositions comprising the same for use in the treatment of cancer. The present invention provides, among ADCs of specific antibodies against insoluble fibrin and drugs, ADCs in which the linker connecting the antibodies and the drugs has a plasmin cleavage sequence, and pharmaceutical compositions for treating cancer comprising the ADCs.)

1. An antibody-drug conjugate (ADC), wherein,

The antibody is an antibody that binds fibrin, and has a higher affinity for insoluble fibrin than for fibrinogen,

The drug is a cytotoxic agent and the drug is a cytotoxic agent,

The antibody and drug are linked in a manner that enables cleavage by plasmin using a linker with a plasmin cleavage site.

2. The ADC of claim 1, wherein the linker has a peptide sequence of valine-leucine-lysine as a plasmin cleavage site.

3. A pharmaceutical composition for treating cancer comprising the ADC of claim 1 or 2.

4. the pharmaceutical composition of claim 3, wherein the cancer is invasive cancer.

5. A fibrin-binding antibody having a heavy chain variable region and a light chain variable region,

The heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 1, CDR1 shown in SEQ ID NO: 2 and CDR2 shown in SEQ ID NO: 3, and a CDR3, and

The light chain variable region has the amino acid sequence shown in SEQ ID NO: 5, CDR1 shown in SEQ ID NO: 6 and the CDR2 shown in SEQ ID NO: the CDR3 shown in figure 7 is,

or an antibody that competes with the antibody for binding to fibrin, or an antigen-binding fragment thereof.

6. An antibody that binds fibrin, or an antigen-binding fragment thereof, said antibody having the amino acid sequence of SEQ ID NO: 4 and SEQ ID NO: 8, or a light chain variable region.

7. a fibrin-binding antibody having a heavy chain variable region and a light chain variable region,

The heavy chain variable region has the amino acid sequence shown in SEQ ID NO: 9, CDR1 shown in SEQ ID NO: 10 and CDR2 shown in SEQ ID NO: 11, and CDR3, and

The light chain variable region has the amino acid sequence shown in SEQ ID NO: 13, CDR1 shown in SEQ ID NO: 14 and CDR2 shown in SEQ ID NO: 15 of the CDR3 shown in figure 15,

or an antibody that competes with the antibody for binding to fibrin, or an antigen-binding fragment thereof.

8. An antibody that binds fibrin, or an antigen-binding fragment thereof, said antibody having the amino acid sequence of SEQ ID NO: 12 and SEQ ID NO: 16, or a light chain variable region as shown in figure 16.

9. an ADC according to claim 1 or 2, wherein the antibody is an antibody according to any one of claims 5 to 8.

10. A pharmaceutical composition comprising the ADC of claim 9.

11. The pharmaceutical composition according to claim 10 for use in the treatment of cancer.

Technical Field

The present invention relates to antibody drug conjugates and compositions comprising the same for the treatment of cancer.

Background

It has been clarified that if there is an angiogenesis injury, blood comes into contact with an injured blood vessel wall and a tissue under the blood vessel, and a tissue factor flows into blood, a coagulation reaction starts, fibrinogen in the blood becomes insoluble fibrin, and a network of fibrin hardens a wound in the form of a firm hemostatic plug.

It has been suggested very early that blood coagulation is closely related to cancer ("edema of thrombus based on limbs in gastric cancer patients") by the surgeon of thuseus (Trousseau) documented in france in the 1800 s. It has been clarified from recent clinical epidemiological data that the frequency of thrombosis caused by a high coagulation rate is significantly higher in almost all cancers, including pancreatic cancer, gastric cancer, and brain tumor (non-patent document 1). In addition, it is considered that coagulation abnormality is also caused in the cancer tissue, and accumulation of insoluble fibrin, coagulation necrosis, and angiogenesis are repeated together with the progress of cancer.

Unlike fibrinogen, which is a precursor, which is widely visible in living bodies, insoluble fibrin is not present in tissues under normal physiological conditions. Insoluble fibrin is produced by polymerizing and crosslinking fibrin monomers, which are formed by cleavage of fibrinogen by thrombin activated by extravasation of the fibrin into blood vessels, to form fibrin fibers. Therefore, insoluble fibrin exists specifically in tissues in pathological states such as bleeding and inflammation, and is formed when a disease accompanied by coagulation such as cancer, myocardial infarction, and cerebral infarction occurs. Therefore, insoluble fibrin is a marker molecule for the disease associated with thrombus, and in particular, insoluble fibrin present in cancer tissues in the absence of cardiovascular and cerebrovascular diseases such as myocardial infarction and cerebral infarction is a cancer-specific molecule.

under such a technical background, an antibody specific to insoluble fibrin and an Antibody Drug Conjugate (ADC) using the same have been proposed (patent document 1).

Disclosure of Invention

The present inventors have developed an ADC having a plasmin cleavage sequence at the linker connecting the antibody and the drug, among specific antibodies against insoluble fibrin and ADCs with the drug. The inventor finds that: the resulting ADC is delivered to insoluble fibrin and is cleaved by plasmin at the site of delivery, where the drug is released. Further, the present inventors have found that, using a tumor model animal, the obtained ADC can target an accumulation site of insoluble fibrin around a tumor, and release a drug at the accumulation site, thereby exerting an anticancer effect against the tumor. In addition, the present inventors obtained novel insoluble fibrin-specific antibodies. The present invention is based on the above findings.

That is, according to the present invention, the following inventions are provided.

(1) An antibody-drug conjugate (ADC), wherein,

The antibody is an antibody that binds fibrin, and has a higher affinity for insoluble fibrin than for fibrinogen,

The drug is a cytotoxic agent and the drug is a cytotoxic agent,

The antibody and drug are linked in a manner that enables cleavage by plasmin using a linker with a plasmin cleavage site.

(2) The ADC of (1) above, wherein,

The linker has a peptide sequence of valine-leucine-lysine as a plasmin cleavage site.

(3) a pharmaceutical composition for treating cancer, which comprises the ADC described in (1) or (2) above.

(4) The pharmaceutical composition of (3) above, wherein the cancer is invasive cancer.

(5) An antibody that binds fibrin, or an antibody that competes with the antibody for binding fibrin, or an antigen-binding fragment thereof,

The antibody that binds fibrin is provided with:

Has the sequence shown in SEQ ID NO: 1, CDR1 shown in SEQ ID NO: 2 and CDR2 shown in SEQ ID NO: 3 of CDR3, and

Has the sequence shown in SEQ ID NO: 5, CDR1 shown in SEQ ID NO: 6 and the CDR2 shown in SEQ ID NO: 7, light chain variable region of CDR 3.

(6) an antibody that binds fibrin, or an antigen-binding fragment thereof, said antibody having the amino acid sequence of SEQ ID NO: 4 and SEQ ID NO: 8, or a light chain variable region.

(7) An antibody that binds fibrin, or an antibody that competes with the antibody for binding fibrin, or an antigen-binding fragment thereof,

the antibody that binds fibrin is provided with:

Has the sequence shown in SEQ ID NO: 9, CDR1 shown in SEQ ID NO: 10 and CDR2 shown in SEQ ID NO: 11 of CDR3, and

Has the sequence shown in SEQ ID NO: 13, CDR1 shown in SEQ ID NO: 14 and CDR2 shown in SEQ ID NO: 15, light chain variable region of CDR 3.

(8) An antibody that binds fibrin, or an antigen-binding fragment thereof, said antibody having the amino acid sequence of SEQ ID NO: 12 and SEQ ID NO: 16, or a light chain variable region as shown in figure 16.

(9) the ADC according to (1) or (2) above, wherein the antibody is the antibody according to any one of (5) to (8) above.

(10) A pharmaceutical composition comprising the ADC described in (9) above.

(11) The pharmaceutical composition of (10) above, which is used for treating cancer.

drawings

FIG. 1 shows that the antibody obtained according to the present invention is an insoluble fibrin-specific antibody.

FIG. 2 shows that the insoluble fibrin-specific monoclonal antibody obtained according to the present invention is accumulated in a tumor-forming part.

FIG. 3 shows in particular the drug site and linker site of the antibody drug conjugate prepared according to the present invention.

Fig. 4 shows the results of the verification of the anticancer effect in vitro using the antibody drug conjugate prepared according to the present invention.

FIG. 5 shows a Kaplan-Meier curve of antibody drug conjugates prepared according to the present invention versus a model of natural pancreatic cancer production.

Fig. 6A shows the inhibitory effect of the antibody drug conjugate prepared according to the present invention on the increase in tumor volume of mice with subcutaneously transplanted tumors.

Fig. 6B shows the change in body weight of the mouse observed in fig. 6A with time.

FIG. 7 shows the cell proliferation inhibitory effect of ADC without plasmin linker containing plasmin cleavage site and plasmin linker, and instead with cathepsin linker containing cathepsin cleavage site.

FIG. 8 shows the putative mechanism of action of antibody drug conjugates prepared according to the present invention.

Detailed Description

In the present invention, the "subject" refers to a mammal, and specifically, a human.

In the present specification, "treatment" is used in a meaning including treatment (therapeutic treatment) and prevention (prophylactic treatment). In the present specification, "treatment" refers to improvement in treatment, cure, prevention, or alleviation of a disease or disorder, or reduction in the rate of progression of a disease or disorder. In the present specification, "prevention" means reducing the possibility of onset of a disease or a condition, or delaying the onset of a disease or a condition.

In the present specification, "disease" refers to a therapeutically useful symptom. In the present specification, "cancer" refers to malignant tumor.

In the present specification, "antibody" refers to immunoglobulin, and includes polyclonal antibodies and monoclonal antibodies. Preferred antibodies are monoclonal antibodies. The source of the antibody is not particularly limited, and examples thereof include: non-human animal antibodies, non-human mammalian antibodies, and human antibodies. The antibody may be a chimeric antibody, a humanized antibody or a human antibody. In addition, the antibody can be a bispecific antibody.

In the present specification, a "therapeutically effective amount" refers to an amount of a pharmaceutical product effective for treating (preventing or treating) a disease or condition. A therapeutically effective amount of the drug is effective to reduce the rate of progression of a disease or condition, halt the progression of the disease, ameliorate the disease, cure the disease, or inhibit the onset or progression of the disease.

In the present specification, "insoluble fibrin" refers to fibrin crosslinked by factor XIII. In a living body, for example, when bleeding occurs, fibrinogen is converted into fibrin monomers by the action of thrombin, and the fibrin monomers are polymerized to form a fibrin polymer that is hardly soluble. The fibrin polymer is cross-linked by factor XIII to become insoluble fibrin.

In the present specification, the term "antibody specific to insoluble fibrin" refers to an antibody that binds to insoluble fibrin and has a higher affinity for insoluble fibrin than for fibrinogen. Such an antibody specific to insoluble fibrin can be easily obtained by screening using the affinity for insoluble fibrin and the affinity for fibrinogen. Fibrin has an epitope site that is converted into insoluble fibrin by a steric conversion and is exposed for the first time. Thus, the "insoluble fibrin-specific antibody" can be obtained by immunizing the exposed domain, i.e., the D domain (hereinafter also referred to as "D-domain"), as an immunogen. In addition, it can also be obtained using a linear peptide. For example, if a partial peptide of a fibrin B.beta.chain corresponding to positions 231 to 246 of the amino acid sequence of the fibrin B.beta.chain (for example, a human fibrin B.beta.chain may have an amino acid sequence represented by SEQ ID NO: 25) is immunized, an "insoluble fibrin-specific antibody" can be obtained. Further, SEQ ID NO: 26 or SEQ ID NO: 27 as an immunogen, or an "insoluble fibrin-specific antibody" can be obtained. Such an antibody specific to insoluble fibrin may be an antibody having a higher affinity for insoluble fibrin than for fibrinogen, fibrin monomers, and fibrin polymers. The ratio of affinity to insoluble fibrin to affinity to fibrinogen may be, for example: antibodies of more than 1, 1.5 or more, 2 or more, 3 or more, 4 or more, or 5 or more are obtained as insoluble fibrin-specific antibodies. Affinity refers to binding affinity (KD), and can be determined by well-known methods such as ELISA and binding equilibrium exclusion.

In this specification, "competition" refers to competition with other binding antibodies for binding to an antigen. For competition, it is possible that there are 2 antibody binding sites that are duplicated for an antigen. Such antibodies can be obtained by immunization using an epitope as described above, and/or can be obtained by confirming whether or not the binding of one antibody to an antigen is reduced by another antibody by a competition assay.

in the present specification, the term "antibody-drug conjugate" (hereinafter also referred to as "ADC") refers to a substance in which an antibody is linked to a cytotoxic agent. In an ADC, the antibody and cytotoxic agent may be linked via an appropriate linker. As the cytotoxic agent, a chemotherapeutic agent, a radioisotope, and a toxin can be used. In ADCs, conjugates of antigen-binding fragments of antibodies with drugs are also included.

In the present specification, the term "antigen-binding fragment" refers to a part of an antibody that retains binding to an antigen. The antigen-binding fragment may comprise the heavy chain variable region or the light chain variable region, or both, of the antibody of the invention. The antigen binding fragment may be chimeric or humanized. Examples of the antigen-binding fragment include: fab, Fab ', F (ab')₂fv, scFv (single-chain Fv), diabody (diabody), sc (Fv)₂(Single chain (Fv)₂). Such a fragment of an antibody is not particularly limited, and can be obtained by treating an antibody with an enzyme, for example. For example, if the antibody is digested with papain, a Fab can be obtained. Alternatively, if the antibody is digested with pepsin, F (ab')₂Further reduction can be carried out to obtain Fab'. In the present invention, an antigen-binding fragment of such an antibody can be used.

in the present invention, in the antibody-drug conjugate, the antibody and the cytotoxic agent are linked via a linker. Examples of the cytotoxic agent include chemotherapeutic agents (e.g., commercially available anticancer agents such as auristatin (auristatin E, auristatin F-phenylenediamine (AFP), monomethyl auristatin E, monomethyl auristatin F and derivatives thereof), maytansine DM1 and DM4 and derivatives thereof), Camptothecin (Camptothecin) (SN-38, irinotecan, Lurtotecan (Lurtotecan), DB67, BMP1350, ST1481, CKD602, topotecan and Exatecan (Exatecan) and derivatives thereof), DNA minor groove binders (enediyne, Lexitropsin, Duocarmycin (Duocarmycin) and derivatives thereof), taxanes (paclitaxel and docetaxel and derivatives thereof), polyketides (polyketilde) (depigment and derivatives thereof), anthraquinones (mitoxantrone and derivatives thereof), benzodiazepines (benzodiazepine and derivatives thereof), benzodiazepines (benzodiazepines and derivatives thereof), and derivatives thereof(Pyrrolobenzodiazepines)IndolesBenzodiazepinesandOxazolidinones benzodiazepinesAnd derivatives thereof), vinca alkaloids (vincristine, vinblastine, vindesine, and vinorelbine and derivatives thereof), doxorubicin (doxorubicin, morpholino-doxorubicin, and cyanomorpholino-doxorubicin and derivatives thereof), cardiac glycosides (digitoxin, derivatives thereof), Calicheamicin (Calicheamicin), epothilone, Cryptophycin (Cryptophycin), cimadrol (Cemadotin), cimadotin (Cemadotin), rhizoxin (rhizoxin), fusin (netropsin), Combretastatin (Combretastatin), punicin (Eleutherobin), etoposide, T67(Tularik), and Nocodazole (Nocodazole)), radioisotopes (e.g.:³²P、⁶⁰C、⁹⁰Y、¹¹¹In、¹³¹I、¹²⁵I、¹⁵³Sm、¹⁸⁶Re、¹⁸⁸Re and²¹²Bi), and toxins (e.g.: diphtheria toxin a, pseudomonas endotoxin, ricin (ricin), Saporin (Saporin), etc.), can be used as the cytotoxic agent in the ADC of the present invention. As cytotoxic agent in the ADC of the present invention, it is preferred to use, for example, camptothecin, in particular SN-38 or irinotecan. As the cytotoxic agent, any of those used in the treatment of cancer can be used. As the cytotoxic agent, a pharmaceutically acceptable salt, solvate (e.g., hydrate), ester or prodrug of the above cytotoxic agent can be used.

In the present invention, the linker of the ADC comprises a plasmin cleavage sequence, capable of being cleaved in the presence of plasmin. In the present invention, the linker of ADC is formed of a stable chemical bond in the process of delivery to insoluble fibrin after administration, except for the plasmin cleavage sequence. By such a constitution, the ADC of the present invention is stable until delivered to insoluble fibrin after administration, and is caused to be cleaved by plasmin after binding to the insoluble fibrin, releasing a cytotoxic agent only in the vicinity of the insoluble fibrin. The plasmin cleavage sequence is an amino acid sequence, and specifically can be a sequence comprising a sequence selected from: peptide chains having an amino acid sequence such as a plasmin cleavage sequence in valine-leucine-lysine, glycine-proline-lysine, glutamic acid-lysine, lysine-phenylalanine-lysine, norvaline-chlorohexylalanyl-lysine, or norleucine-hexahydrotyrosine-lysine. Such a linker can be appropriately selected and synthesized by those skilled in the art in the preparation of ADCs. In one embodiment, for the joint, for example: a1 st spacer may be introduced between the antibody and the plasmin cleavage sequence, and polyethylene glycol (PEG), for example, having about 5 to 40 repeating units per molecule may be used as the 1 st spacer. A2 nd spacer may also be introduced between the plasmin cleavage sequence and the cytotoxic agent, and for example, p-aminobenzyloxycarbonyl (PABC) may be used as the 2 nd spacer.

In one embodiment, the linker comprises a 1 st spacer and a plasmin cleavage sequence. In certain embodiments, the linker comprises a 1 st spacer, a plasmin cleavage sequence, and a 2 nd spacer. In a particular subject, the linker comprises PEG, a plasmin cleavage sequence, and a PABC.

In certain embodiments, the linker does not comprise a cleavage moiety other than a plasmin cleavage sequence.

the binding of the antibody to the linker may be performed, for example, by linking via a maleimide group on a thiol group of the antibody.

in certain embodiments, the antibody is linked to the anti-cancer agent via the thiol group using a linker having a maleimide-PEG-plasmin cleavage sequence. In one embodiment, the antibody is linked to the anti-cancer agent via the thiol group using a linker having a maleimide-PEG-plasmin cleavage sequence-PABC.

In any of the ADCs of the present invention, the anticancer agent is linked to the anticancer agent by a linker having a plasmin cleavage site that can be cleaved by plasmin, and when the ADC reaches a site where insoluble fibrin accumulates, the linker is cleaved at the plasmin cleavage site by plasmin present around the ADC, thereby releasing the anticancer agent around the insoluble fibrin. It is considered that there are many sites around cancer tissues where insoluble fibrin accumulates due to bleeding caused by cancer infiltration (see fig. 8), and the antibody of the present invention (i.e., an antibody specific to insoluble fibrin) is useful for targeting cancer in a drug delivery system, and the ADC of the present invention is useful as a therapeutic agent for cancer.

the following antibodies are provided in the present invention:

An antibody that binds fibrin, and an antibody that competes with the antibody for binding to fibrin,

The antibody that binds fibrin is provided with:

Has the sequence shown in SEQ ID NO: 1, CDR1 shown in SEQ ID NO: 2 and CDR2 shown in SEQ ID NO: 3 of CDR3, and

Has the sequence shown in SEQ ID NO: 5, CDR1 shown in SEQ ID NO: 6 and the CDR2 shown in SEQ ID NO: 7, light chain variable region of CDR 3. These antibodies can be used as insoluble fibrin-specific antibodies.

In the present invention, the following antibodies are additionally provided:

A fibrin-binding antibody having the amino acid sequence of SEQ ID NO: 4 and SEQ ID NO: 8 in the variable region of the light chain. The antibody can be used as an antibody specific to insoluble fibrin.

The antibody can be said to have:

Has the sequence shown in SEQ ID NO: 1, CDR1 shown in SEQ ID NO: 2 and CDR2 shown in SEQ ID NO: 3, and a CDR3 heavy chain variable region, and

has the sequence shown in SEQ ID NO: 5, CDR1 shown in SEQ ID NO: 6 and the CDR2 shown in SEQ ID NO: 7, and an antibody comprising the light chain variable region of CDR 3.

In the present invention, the following antibodies are provided:

An antibody that binds fibrin, and an antibody that competes with the antibody for binding to fibrin,

The antibody that binds fibrin is provided with:

Has the sequence shown in SEQ ID NO: 9, CDR1 shown in SEQ ID NO: 10 and CDR2 shown in SEQ ID NO: 11 of CDR3, and

Has the sequence shown in SEQ ID NO: 13, CDR1 shown in SEQ ID NO: 14 and CDR2 shown in SEQ ID NO: 15, light chain variable region of CDR 3. These antibodies can be used as insoluble fibrin-specific antibodies.

in the present invention, the following antibodies are additionally provided:

A fibrin-binding antibody having the amino acid sequence of SEQ ID NO: 12 and SEQ ID NO: 16 in the variable region of the light chain. The antibody can be used as an antibody specific to insoluble fibrin.

The antibody can be said to have:

Has the sequence shown in SEQ ID NO: 9, CDR1 shown in SEQ ID NO: 10 and CDR2 shown in SEQ ID NO: 11 of CDR3, and

Has the sequence shown in SEQ ID NO: 13, CDR1 shown in SEQ ID NO: 14 and CDR2 shown in SEQ ID NO: 15 of CDR 3.

The above-mentioned antibodies, insoluble fibrin-specific antibodies, and antigen-binding fragments thereof can be used as the antibody portion in the ADC of the present invention.

According to the present invention there is provided a pharmaceutical composition comprising a therapeutically effective amount of an ADC as described above (also referred to as "ADC of the invention"). According to the present invention, the ADC of the present invention and the above pharmaceutical composition, respectively, can be used for the treatment of cancer.

The cancer to be treated by the ADC or the pharmaceutical composition of the present invention is not particularly limited, and examples thereof include: lung cancer, pancreatic cancer, head and neck cancer, prostate cancer, bladder cancer, breast cancer, esophageal cancer, stomach cancer, colorectal cancer, uterine cancer, ovarian cancer, skin cancer, thyroid cancer, thymus cancer, kidney cancer, testicular cancer, penile cancer, liver cancer, bile duct cancer, brain tumor, bone and soft tissue tumor, retroperitoneal tumor, angiolymphangiosarcoma, and metastatic cancers thereof.

The subject of the present invention may be a subject who does not suffer from a thrombotic disorder or a disease associated with a thrombotic disorder, or a subject who has not been diagnosed as a thrombotic disorder or a disease associated with a thrombotic disorder. This is expected to reduce side effects on tissues other than cancer. Thus, the ADC of the present invention may also be administered to a subject who does not suffer from a thrombotic disorder or a disease associated with a thrombotic disorder after determining whether the subject suffers from a thrombotic disorder or a disease associated with a thrombotic disorder to a subject who suffers from cancer. Whether or not a thrombotic disorder or a disease associated with a thrombotic disorder is present can be appropriately judged by a doctor.

In a certain embodiment of the invention, the pharmaceutical composition comprises an ADC of the invention and an excipient. The pharmaceutical composition of the invention can be prepared by: intravenous administration, subcutaneous administration, intratumoral administration, intraperitoneal administration, intracerebroventricular administration, intramuscular administration, and the like. The amount can be appropriately determined by a physician in consideration of the age, sex, weight, severity of the disease, etc. of the patient.

The ADC of the present invention targets insoluble fibrin accumulated in the interstitium of these cancers to accumulate a cytotoxic agent at the targeted site, and further has a linker that is activated at the site where the insoluble fibrin exists and is cleavable by plasmin, and releases the cytotoxic agent at the targeted site. This can cause site-specific damage to cancer around the free site.

according to the present invention there is provided the use of an insoluble fibrin-specific antibody in the manufacture of a medicament for the treatment of cancer. According to the present invention there is provided the use of an ADC, which is an ADC of an insoluble fibrin-specific antibody and a cytotoxic agent having a plasmin cleavage site capable of being cleaved by plasmin, in the manufacture of a medicament for the treatment of cancer.

According to the present invention there is provided a method of treating cancer in a subject in need thereof, the method comprising: administering to the subject a therapeutically effective amount of an ADC of the invention. According to the present invention there is provided a method of treating cancer in a subject in need thereof, comprising: after determining whether a subject having cancer has a thrombotic disorder or a disease associated with a thrombotic disorder, a therapeutically effective amount of an ADC of the invention is administered to a subject not having a thrombotic disorder or a disease associated with a thrombotic disorder.

According to the invention there is provided the use of an ADC of the invention for use in a method of treating cancer.

examples