Cell penetrating nucleolytic antibody based cancer therapy

Abstract

Cancer cells with defects in DNA repair are highly susceptible to DNA-damaging agents, but delivery of therapeutic agents into cell nuclei can be challenging. A sub-set of autoantibodies having nucleolytic activity are capable of nuclear penetration. These antibodies can be used as therapeutic agents targeted towards DNA repair-deficient malignancies.

1 .- 15 . (canceled) 
     
     
         16 . A method of treating cancer, comprising
 administering to a subject in need thereof a pharmaceutical composition comprising a cell-penetrating, nucleolytic antibody, or antigen binding fragment thereof, comprising   a heavy chain variable region comprising respectively the first, second, and third complementarity determining regions (CDRs) of SEQ ID NO:5 or humanized variants thereof, and   a light chain variable region comprising respectively the first, second, and third CDRs of SEQ ID NO:1 or humanized variants thereof.   
     
     
         17 . The method of  claim 16 , wherein the pharmaceutical composition is in a unit dosage between about 0.01 to about 100 mg/kg body weight of a human. 
     
     
         18 . The method of  claim 16 , wherein the cancer is selected from the group consisting of breast cancers, colon cancers, endometrial tumors, brain tumors, ovarian, and pancreatic cancers, leukemias and other cancers of the blood and lymphatic system, cancers of the genitourinary system, cancers of the nervous system, cancers of the head and neck, lung cancers, gynecologic cancers, gastrointestinal cancers, skin cancers, and pediatric cancers. 
     
     
         19 . The method of  claim 16 , wherein the cancer is characterized by intrinsic deficiency or deficiencies in DNA repair. 
     
     
         20 . The method of  claim 16 , wherein the subject that has been diagnosed with one or more mutations in one or more DNA repair genes selected from the group consisting of ATM, ATR, BRCA1, BRCA2, FANCD2, MLH1, MRE11, MSH2, PALB2, PMS2 and PTEN. 
     
     
         21 . The method of  claim 16 , further comprising treating the subject with radiation therapy, chemotherapy, or combinations therefor,
 wherein the cell-penetrating nucleolytic antibody increases the cell's sensitivity to radiation therapy, chemotherapy, or combinations thereof by at least 10%.   
     
     
         22 .- 23 . (canceled) 
     
     
         24 . The method of  claim 16 , wherein the pharmaceutical composition further comprises one or more antineoplastic or radio-sensitizing agents selected from the group consisting of cisplatin, cytoxan, doxorubicin, methotrexate, mitomycin c, nitrogen mustard, hydroxyurea, bevacizumab, cetuximab, rituximab, trastuzumab, tirapazamine, temozolomide, camptothecin, cisplatin, gemcitabine, 5-fluorouracil, hydroxyurea, pentoxifylline, vinorelbine, and combinations thereof. 
     
     
         25 . The method of  claim 16 , wherein the composition is administered via intravenous injection or intratumoral injection. 
     
     
         26 . The method of  claim 16 , wherein the antibody or antigen binding fragment thereof in the composition is a single chain variable fragment (scFv) or a di-scFv. 
     
     
         27 . The method of  claim 16 , wherein the antibody or antigen binding fragment thereof in the composition comprises:
 a first light chain CDR which comprises the amino acid sequence of SEQ ID NO:2;   a second light chain CDR which comprises the amino acid sequence of SEQ ID NO:3;   a third light chain CDR which comprises the amino acid sequence of SEQ ID NO:4;   a first heavy chain CDR which comprises the amino acid sequence of SEQ ID NO:6;   a second heavy chain CDR which comprises the amino acid sequence of SEQ ID NO:7; and   a third heavy chain CDR which comprises the amino acid sequence of SEQ ID NO:8.   
     
     
         28 . The method of  claim 16 , wherein the antibody or antigen binding fragment thereof in the composition comprises the amino acid sequence of SEQ ID NO: 1 and the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 5. 
     
     
         29 . The method of  claim 16 , wherein the antibody or antigen binding fragment thereof in the composition is a humanized antibody or an antigen binding fragment thereof. 
     
     
         30 . The method of  claim 29 , wherein the humanized antibody or antigen binding fragment thereof in the composition comprises:
 a first light chain CDR which comprises the amino acid sequence of SEQ ID NO:2;   a second light chain CDR which comprises the amino acid sequence of SEQ ID NO:3;   a third light chain CDR which comprises the amino acid sequence of SEQ ID NO:4;   a first heavy chain CDR which comprises the amino acid sequence of SEQ ID NO:6;   a second heavy chain CDR which comprises the amino acid sequence of SEQ ID NO:7; and   a third heavy chain CDR which comprises the amino acid sequence of SEQ ID NO:8.   
     
     
         31 . A pharmaceutical composition comprising
 a) a cell-penetrating, nucleolytic antibody, or antigen binding fragment thereof, comprising:   a heavy chain variable region comprising respectively the first, second, and third complementarity determining regions (CDRs) of SEQ ID NO:5 or humanized variants thereof, and   a light chain variable region comprising respectively the first, second, and third CDRs of SEQ ID NO:1 or humanized variants thereof; and   b) a pharmaceutically acceptable excipient for injection.   
     
     
         32 . The pharmaceutical composition of  claim 31 , wherein the antibody or antigen binding fragment or fusion or variant thereof is present in amount from about 0.01 to about 100 mg/kg body weight of a human. 
     
     
         33 . A dosage unit formulation for administration to a patient with solid tumors comprising a therapeutically effective amount of the composition of  claim 31 . 
     
     
         34 . The dosage unit formulation of  claim 33  wherein the antibody is in lyophilized or nanoparticulate, microparticulate, liposomal or precipitated form.