Targeting tumor cells with chemotherapeutic agents conjugated to anti-matriptase antibodies by in vivo cleavable linking moieties

Abstract

The present invention relates to anti-matriptase antibodies and immunoconjugates of anti-matriptase antibodies with cytotoxic agents and the use thereof for killing or inhibiting the growth of matriptase-expressing cancer cells, such as those of multiple myeloma and breast cancers. In particular, immunoconjugates comprising an anti-matriptase monoclonal antibody and anticancer agents such as auristatin, including monomethyl auristatin E (MMAE) and monomethyl auristatin F (MMAF) are introduced, which have potent antitumor activity in vivo. Moreover, importantly; there was no weight loss or other evidence of toxicity in the animals, indicating that no significant free drug was released into the circulation from the conjugate. The present invention also provides compositions comprising these new immunoconjugates and use of them for treatment of malignancies comprising cells that express matriptase. In addition, administration of an anti-matriptase antibody or immunoconjugates of an anti-matriptase antibody and a cytotoxic agent in combination with administration of an immunomodulatory agent, such as thalidomide or an analog thereof, provides a more effective treatment of these cancers.

1 . An immunoconjugate selectively targeting cancer cells that express matriptase, comprising an anti-matriptase antibody or antigen-binding fragment thereof, and a cytotoxic agent, wherein said cytotoxic agent is selected from the group consisting of monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), and auristatin PE, wherein the anti-matriptase antibody or antigen-binding fragment thereof is specific for activated matriptase. 
     
     
         2 . The immunoconjugate of  claim 1 , wherein the anti-matriptase antibody comprises M69 monoclonal antibody or antigen-binding portion thereof. 
     
     
         3 . The immunoconjugate of  claim 1 , wherein the immunoconjugate further comprises a linker comprising a cleavable linking moiety. 
     
     
         4 . (canceled) 
     
     
         5 . The immunoconjugate of  claim 3 , wherein the cleavable linking moiety comprises a Val-Cit linking moiety or a Phy-Lys linking moiety, cleavable by Capthesin B. 
     
     
         6 . The immunoconjugate of  claim 1 , wherein the linker is covalently bound to a lysine residue on the anti-matriptase antibody. 
     
     
         7 . The immunoconjugate of  claim 1 , wherein the linker is PEG-containing. 
     
     
         8 . The immunoconjugate of  claim 1 , wherein the immunoconjugate further comprises a linker, and the linker comprises a first linking component and a second linking component. 
     
     
         9 . The immunoconjugate of  claim 8 , wherein the first linking component is covalently bound to a lysine residue on the anti-matriptase antibody. 
     
     
         10 . The immunoconjugate of  claim 8 , wherein the first linking component is bound to the second linking component through a triazole moiety. 
     
     
         11 . The immunoconjugate of  claim 8 , wherein the second linking component comprises a cleavable linking moiety. 
     
     
         12 . The immunoconjugate of  claim 11 , wherein the cleavable linking moiety comprises a Val-Cit linking moiety or a Phy-Lys linking moiety, cleavable by Capthesin B. 
     
     
         13 . The immunoconjugate of  claim 8 , wherein at least one of the first linking component and the second linking component is PEG-containing. 
     
     
         14 . (canceled) 
     
     
         15 . The immunoconjugate of  claim 8 , wherein the cytotoxic agent is bound to the second linking component. 
     
     
         16 . A method of treating a malignancy comprising cells that express matriptase, the method comprising administering to a subject in need of such a treatment a therapeutically effective amount of a composition comprising the immunoconjugate of  claim 1 . 
     
     
         17 . The method of  claim 16  wherein the malignancy comprises a hematological malignancy selected from the group consisting of acute lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), chronic myelogenous leukemia (CML), acute monocytic leukemia (AMOL)), Hodgkin's lymphomas, Non-Hodgkin's lymphomas, Burkitt's lymphoma (BL), diffuse large B-cell lymphoma (DLBL), Mantle cell lymphoma (MCL), multiple myeloma (MM), and an epithelial malignancy selected from the group consisting of prostate, breast, brain, kidney, lung, colon, bladder, skin, thyroid, ovary tumors, and mesothelioma. 
     
     
         18 . (canceled) 
     
     
         19 . (canceled) 
     
     
         20 . (canceled) 
     
     
         21 . The method of  claim 16 , further comprising co-administering a therapeutically effective amount of an immunomodulatory agent that activates matriptase. 
     
     
         22 . The method of  claim 21 , wherein the immunomodulatory agent comprises thalidomide or a thalidomide analog. 
     
     
         23 . An immunoconjugate comprising an anti-matriptase antibody or an antigen-binding portion thereof that is specific for activated matriptase, covalently bound to a cytotoxic agent through a linker, the cytotoxic agent being selected from the group consisting of monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), and auristatin PE, and the linker comprising:
 a first linking component bound to the anti-matriptase antibody or an antigen-binding portion thereof through a lysine residue located on the anti-matriptase antibody or an antigen-binding portion thereof; and   a second linking component comprising a cleavable linking moiety, the second linking component being bound to the first linking component through a triazole moiety and to the cytotoxic agent through the cleavable linking moiety,   wherein the cleavable linking moiety comprises a Val-Cit linking moiety or a Phy-Lys linking moiety, cleavable by Capthesin B,   wherein the first linking component comprises between about 3 and about 7 poly(ethylene glycol) (PEG) units between the amide bond between the lysine residue on the anti-matriptase antibody or an antigen-binding portion thereof and the triazole moiety, and   wherein the second linking component comprises between about 3 and about 7 poly(ethylene glycol) (PEG) units between the triazole moiety and the cleavable linking moiety.   
     
     
         24 . The immunoconjugate of  claim 23 , wherein the anti-matriptase antibody or an antigen-binding portion thereof comprises M69 or an antigen-binding portion thereof. 
     
     
         25 . The immunoconjugate of  claim 24 , wherein the cytotoxic agent comprises monomethyl auristatin E (MMAE).