Anti-EGFR antibody and uses of same
This disclosure relates generally to an EGFR antibody and its therapeutic effects on tumor inhibition in vitro and in vivo, alone or in combination with various chemotherapeutic agents. In particular, the present disclosure relates to methods for the treatment of cancer, comprising administering an EGFR antibody, alone or in combination with a chemotherapeutic agent.
1. An antibody which binds epidermal growth factor receptor (EGFR) comprising a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:2 in combination with a light chain consisting of the amino acid sequence set forth in SEQ ID NO:1, wherein the antibody comprises N-acetylneuraminic acid (NANA) and lacks galactose-α-1,3-galactose, wherein the antibody comprises disulfide bonds between amino acid residues:
Cys23 and Cys88 of SEQ ID NO:1;
Cys134 and Cys184 of SEQ ID NO:1;
Cys214 of SEQ ID NO: 1 and Cys225 of SEQ ID NO: 2;
Cys22 and Cys95 of SEQ ID NO:2;
Cys146 and Cys202 of SEQ ID NO:2;
Cys266 and Cys326 of SEQ ID NO:2; and
Cys372 and Cys430 of SEQ ID NO:2;
wherein SEQ ID NO: 2 comprises N-glycan at amino acid residues Asn88 and Asn302; and
wherein the N-glycan at amino acid residue Asn88 comprises NANA.
2. The antibody of claim 1 , wherein the antibody is conjugated to a detectable marker.
3. The antibody of claim 2 , wherein the detectable marker comprises a radionuclide or a fluorescent label.
4. The antibody of claim 1 , wherein the antibody is conjugated to one or more additional therapeutic agents.
5. The antibody of claim 4 , wherein:
a) the additional therapeutic agent is selected from the group consisting of a vinca alkaloid, a microtubule disrupting agent, an anti-angiogenic agent, and a therapeutic antibody; or wherein
b) the additional therapeutic agent is selected from the group consisting of an EGFR targeting agent, a tyrosine kinase targeting agent, a transitional metal complex, a proteasome inhibitor an antimetabolite, an alkylating agent, a platinum-based agent, an anthracycline antibiotic, a topoisomerase inhibitor, a macrolide, and a retinoid; or wherein
c) the additional therapeutic agent is selected from the group consisting of geldanamycin or a derivative thereof, adriamycin, colchicine, cyclophosphamide, actinomycin, bleomycin, duanorubicin, doxorubicin, epirubicin, mitomycin, methotrexate, mitoxantrone, fluorouracil, carboplatin, carmustine (BCNU), methyl-CCNU, cisplatin, etoposide, interferons, camptothecin and derivatives thereof, phenesterine, taxanes and derivatives thereof, topetecan, vinblastine, vincristine, tamoxifen, piposulfan, nab-5404, nab-5800, and nab-5801; or wherein
d) the additional therapeutic agent is selected from the group consisting Irinotecan, HKP, Ortataxel, gemcitabine, Oxaliplatin, Herceptin®, vinorelbine, Doxil®, capecitabine, Alimta®, Avastin®, Velcade®, Tarceva®, Neulasta®, lapatinib, and sorafenib.
6. A composition comprising the antibody of claim 1 , wherein the antibody has a higher degree of thermostability than Erbutix® (cetuximab), and wherein the antibody does not induce a hypersensitivity response in a subject hypersensitive to cetuximab or predisposed to having a hypersensitivity reaction to cetuximab.
7. The composition of claim 6 , further comprising a chemotherapeutic agent, wherein:
a) the chemotherapeutic agent is selected from the group consisting of a vinca alkaloid, a microtubule disrupting agent, an anti-angiogenic agent, and a therapeutic antibody; or wherein
b) the chemotherapeutic agent is selected from the group consisting of an EGFR targeting agent, a tyrosine kinase targeting agent, a transitional metal complex, a proteasome inhibitor, an antimetabolite, an alkylating agent, a platinum-based agent, an anthracycline antibiotic, a topoisomerase inhibitor, a macrolide, and a retinoid; or wherein
c) the chemotherapeutic agent is selected from the group consisting of geldanamycin or a derivative thereof, adriamycin, colchicine, cyclophosphamide, actinomycin, bleomycin, duanorubicin, doxorubicin, epirubicin, mitomycin, methotrexate, mitoxantrone, fluorouracil, carboplatin, carmustine (BCNU), methyl-CCNU, cisplatin, etoposide, interferons, camptothecin and derivatives thereof, phenesterine, taxanes and derivatives thereof, topetecan, vinblastine, vincristine, tamoxifen, piposulfan, nab-5404, nab-5800, and nab-5801; or wherein
d) the chemotherapeutic agent is selected from the group consisting of Irinotecan, HKP, Ortataxel, gemcitabine, Oxaliplatin, Herceptin®, vinorelbine, Doxil®, capecitabine, Alimta®, Avastin®, Velcade®, Tarceva®, Neulasta®, lapatinib, and sorafenib.
8. The antibody of claim 1 , wherein the antibody has a higher degree of thermostability than Erbutix® (cetuximab), and wherein the antibody does not induce a hypersensitivity response in a subject hypersensitive to cetuximab or predisposed to having a hypersensitivity reaction to cetuximab.
9. A Chinese hamster ovary (CHO) cell comprising polynucleotides encoding the antibody of claim 1 .
10. The CHO cell of claim 9 , wherein the nucleic acids are present on a replicable vector separate from the CHO cell genome.
11. The CHO cell of claim 9 , wherein the nucleic acids are stably integrated into the CHO cell genome.
12. A method of producing the antibody of claim 1 , comprising introducing into a Chinese hamster ovary (CHO) cell with polynucleotides encoding the amino acid sequences set forth in SEQ ID NO: 1 and SEQ ID NO: 2, wherein the CHO cell subsequently expresses the nucleic acid sequences and produces an antibody comprising a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 1 in combination with a light chain consisting of the amino acid sequence set forth in SEQ ID NO: 2.
13. A method for treating an EGFR-expressing cancer in a subject in need thereof, comprising administering to the subject the antibody of claim 1 , and further comprising determining whether the subject is hypersensitive to cetuximab or is predisposed to having a hypersensitivity reaction to cetuximab, wherein the antibody has a higher degree of thermostability than Erbutix® (cetuximab).
14. The method of claim 13 , further comprising administering one or more additional therapeutic agents, wherein:
a) the additional therapeutic agent is selected from the group consisting of a vinca alkaloid, a microtubule disrupting agent, an anti-angiogenic agent, and a therapeutic antibody; or wherein
b) the additional therapeutic agent is selected from the group consisting of an EGFR targeting agent, a tyrosine kinase targeting agent, a transitional metal complex, a proteasome inhibitor, an antimetabolite, an alkylating agent, a platinum-based agent, an anthracycline antibiotic, a topoisomerase inhibitor, a macrolide, and a retinoid; or wherein
c) the additional therapeutic agent is selected from the group consisting of geldanamycin or a derivative thereof, adriamycin, colchicine, cyclophosphamide, actinomycin, bleomycin, duanorubicin, doxorubicin, epirubicin, mitomycin, methotrexate, mitoxantrone, fluorouracil, carboplatin, carmustine (BCNU), methyl-CCNU, cisplatin, etoposide, interferons, camptothecin and derivatives thereof, phenesterine, taxanes and derivatives thereof, topetecan, vinblastine, vincristine, tamoxifen, piposulfan, nab-5404, nab-5800, and nab-5801; or wherein
d) the additional therapeutic agent is selected from the group consisting of Irinotecan, HKP, Ortataxel, gemcitabine, Oxaliplatin, Herceptin®, vinorelbine, Doxil®, capecitabine, Alimta®, Avastin®, Velcade®, Tarceva®, Neulasta®, lapatinib, and sorafenib.
15. The method of claim 13 , wherein the antibody is conjugated to one or more additional therapeutic agents.
16. The method of claim 13 , wherein the subject has elevated levels of anti-cetuximab IgE compared to a negative control sample.
17. The method of claim 13 , wherein the subject has elevated levels of anti-galactose-α-1,3-galactose IgE compared to a negative control sample.
18. The method of claim 13 , wherein determining comprises measuring the presence of anti-cetuximab or anti-galactose-α-1,3-galactose IgE in a sample of serum from the subject, wherein an elevated level of anti-cetuximab or anti-galactose-α-1,3-galactose IgE compared to a negative control sample indicates that the subject is hypersensitive to cetuximab or is predisposed to having a hypersensitivity reaction to cetuximab.