Anti-cd33 antibodies and method for treatment of acute myeloid leukemia using the same

Abstract

The present invention relates to antibodies that bind CD33. More particularly, the invention relates to anti-CD33 antibodies, fragments and homologues of these antibodies, humanized and resurfaced versions of these antibodies, functional equivalents and improved versions of these antibodies, immunoconjugates and compositions comprising these antibodies, and the uses of same in diagnostic, research and therapeutic applications. The invention also relates to a polynucleotide encoding these antibodies, vectors comprising the polynucleotides, host cells transformed with polynucleotides and methods of producing these antibodies.

1 . An isolated antibody or fragment thereof, comprising at least one complementarity-determining region having an amino acid sequence selected from the group consisting of SEQ ID NOs:1-6. 
     
     
         2 . An antibody or fragment thereof, comprising at least one heavy chain variable region and at least one light chain variable region, wherein said heavy chain variable region comprises at least one complementarity-determining region having the amino acid sequence of SEQ ID NOs:1, 2 or 3, and wherein said light chain variable region comprises at least one complementarity-determining region having the amino acid sequences represented by SEQ ID NOs:4, 5 or 6. 
     
     
         3 . The antibody or fragment thereof of  claim 2 , wherein said heavy chain variable region has at least 90% sequence identity to an amino acid sequence represented by SEQ ID NO:7. 
     
     
         4 . The antibody or fragment thereof of  claim 2 , wherein said heavy chain variable region has at least 95% sequence identity to said amino acid sequence represented by SEQ ID NO:7. 
     
     
         5 . (canceled) 
     
     
         6 . The antibody or fragment thereof of  claim 2 , wherein said light chain variable region has at least 90% sequence identity to an amino acid sequence represented by SEQ ID NO:8. 
     
     
         7 . The antibody or fragment thereof of  claim 2 , wherein said light chain variable region has at least 95% sequence identity to said amino acid sequence represented by SEQ ID NO:8. 
     
     
         8 . (canceled) 
     
     
         9 . The antibody or fragment thereof of  claim 2 , wherein said heavy chain variable region has at least 90% sequence identity to an amino acid sequence represented by SEQ ID NO:9. 
     
     
         10 . The antibody or fragment thereof of  claim 2 , wherein said heavy chain variable region has at least 95% sequence identity to said amino acid sequence represented by SEQ ID NO:9. 
     
     
         11 . (canceled) 
     
     
         12 . The antibody or fragment thereof of  claim 2 , wherein said light chain variable region has at least 90% sequence identity to an amino acid sequence represented by SEQ ID NO:10. 
     
     
         13 . The antibody or fragment thereof of  claim 2 , wherein said light chain variable region has at least 95% sequence identity to said amino acid sequence represented by SEQ ID NO:10. 
     
     
         14 - 16 . (canceled) 
     
     
         17 . An immunoconjugate comprising the antibody or epitope-binding fragment thereof of  claim 1  linked to a drug or prodrug. 
     
     
         18 . An immunoconjugate comprising the antibody or epitope-binding fragment thereof of  claim 2  linked to a drug or prodrug. 
     
     
         19 . The immunoconjugate of  claim 17 , wherein said drug or prodrug is selected from the group consisting of a maytansinoid, a taxoid, CC-1065, a CC-1065 analog, dolastatin, a dolastatin analog, methotrexate, daunorubicin, doxorubicin, vincristine, vinblastine, melphalan, mitomycin C, chlorambucil, calicheamicin, and derivatives thereof. 
     
     
         20 . The immunoconjugate of  claim 18 , wherein said drug or prodrug is selected from the group consisting of a maytansinoid, a taxoid, CC-1065, a CC-1065 analog, dolastatin, a dolastatin analog, methotrexate, daunorubicin, doxorubicin, vincristine, vinblastine, melphalan, mitomycin C, chlorambucil, calicheamicin, and derivatives thereof. 
     
     
         21 . A composition comprising the antibody or epitope-binding fragment thereof of  claim 1  and a drug or prodrug. 
     
     
         22 . A composition comprising the antibody or epitope-binding fragment thereof of  claim 2  and a drug or prodrug. 
     
     
         23 . A pharmaceutical composition comprising the antibody or epitope-binding fragment thereof of  claim 1 , and a pharmaceutically acceptable agent. 
     
     
         24 . A pharmaceutical composition comprising the antibody or epitope-binding fragment thereof of  claim 2 , and a pharmaceutically acceptable agent. 
     
     
         25 . A pharmaceutical composition comprising the immunoconjugate of  claim 17 , and a pharmaceutically acceptable agent. 
     
     
         26 . A pharmaceutical composition comprising the immunoconjugate of  claim 18 , and a pharmaceutically acceptable agent. 
     
     
         27 . A pharmaceutical composition comprising the composition of  claim 21 , and a pharmaceutically acceptable agent. 
     
     
         28 . A pharmaceutical composition comprising the composition of  claim 22 , and a pharmaceutically acceptable agent. 
     
     
         29 . A diagnostic reagent comprising the antibody of  claim 1 , wherein said antibody or antibody fragment is labeled. 
     
     
         30 . A diagnostic reagent comprising the antibody of  claim 2 , wherein said antibody or antibody fragment is labeled. 
     
     
         31 . The diagnostic reagent of  claim 29 , wherein said label is selected from the group consisting of a biotin label, an enzyme label, a radio-label, a fluorophore, a chromophore, an imaging agent and a metal ion. 
     
     
         32 . The diagnostic reagent of  claim 30 , wherein said label is selected from the group consisting of a biotin label, an enzyme label, a radio-label, a fluorophore, a chromophore, an imaging agent and a metal ion. 
     
     
         33 . A method for inhibiting the growth of a cell expressing CD33 comprising contacting said cell with the antibody or epitope-binding fragment thereof of  claim 1  or  2 . 
     
     
         34 . A method for inhibiting the growth of a cell expressing CD33 comprising contacting said cell with the immunoconjugate of  claim 17  or  18 . 
     
     
         35 . A method for inhibiting the growth of a cell expressing CD33 comprising contacting said cell with the composition of  claim 21  or  22 . 
     
     
         36 . A method for inhibiting the growth of a cell expressing CD33 comprising contacting said cell with a pharmaceutical composition selected from  claims 23 - 28 . 
     
     
         37 . A method for treating a subject having a disease wherein CD33 is expressed, comprising administering to said subject an effective amount of the antibody or epitope-binding fragment thereof of  claim 1  or  2 . 
     
     
         38 . A method for treating a subject having a disease wherein CD33 is expressed, comprising administering to said subject an effective amount of the immunoconjugate of  claim 17  or  18 . 
     
     
         39 . A method for treating a subject having a disease wherein CD33 is expressed, comprising administering to said subject an effective amount of the composition of  claim 21  or  22 . 
     
     
         40 . A method for treating a subject having a disease wherein CD33 is expressed, comprising administering to said subject an effective amount of the pharmaceutical composition of  claim 23  or  24 . 
     
     
         41 . A method for treating a subject having a disease wherein CD33 is expressed, comprising administering to said subject an effective amount of the pharmaceutical composition of  claim 25  or  26 . 
     
     
         42 . A method for treating a subject having a disease wherein CD33 is expressed, comprising administering to said subject an effective amount of the pharmaceutical composition of  claim 27  or  28 . 
     
     
         43 . A method for treating a subject having a disease wherein CD33 is expressed, comprising contacting one or more cells of said subject ex vivo with an effective amount of the antibody or epitope-binding fragment thereof of  claim 1  or  2 . 
     
     
         44 . A method for treating a subject having a disease wherein CD33 is expressed, comprising contacting one or more cells of said subject ex vivo with an effective amount of an immunoconjugate of  claim 17  or  18 . 
     
     
         45 . A method for treating a subject having a disease wherein CD33 is expressed, comprising contacting one or more cells of said subject ex vivo with an effective amount of a composition of  claim 21  or  22 . 
     
     
         46 . A method for treating a subject having a disease wherein CD33 is expressed, comprising contacting one or more cells of said subject ex vivo with an effective amount of a pharmaceutical composition selected from  claims 23 - 28 . 
     
     
         47 . The method of treatment of  claim 37 , wherein said disease is a disease selected from the group consisting of myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), chronic myeloid leukemia (CML) and pro-myelocytic leukemia (PML). 
     
     
         48 . The method of treatment of  claim 38 , wherein said disease is a disease selected from the group consisting of myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), chronic myeloid leukemia (CML) and pro-myelocytic leukemia (PML). 
     
     
         49 . The method of treatment of  claim 39 , wherein said disease is a disease selected from the group consisting of myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), chronic myeloid leukemia (CML) and pro-myelocytic leukemia (PML). 
     
     
         50 . The method of treatment of  claim 40 , wherein said disease is a disease selected from the group consisting of myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), chronic myeloid leukemia (CML) and pro-myelocytic leukemia (PML). 
     
     
         51 . The method of treatment of  claim 41 , wherein said disease is a disease selected from the group consisting of myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), chronic myeloid leukemia (CML) and pro-myelocytic leukemia (PML). 
     
     
         52 . The method of treatment of  claim 42 , wherein said disease is a disease selected from the group consisting of myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), chronic myeloid leukemia (CML) and pro-myelocytic leukemia (PML). 
     
     
         53 . The method of treatment of  claim 43 , wherein said disease is a disease selected from the group consisting of myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), chronic myeloid leukemia (CML) and pro-myelocytic leukemia (PML). 
     
     
         54 . The method of treatment of  claim 44 , wherein said disease is a disease selected from the group consisting of myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), chronic myeloid leukemia (CML) and pro-myelocytic leukemia (PML). 
     
     
         55 . The method of treatment of  claim 45 , wherein said disease is a disease selected from the group consisting of myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), chronic myeloid leukemia (CML) and pro-myelocytic leukemia (PML). 
     
     
         56 . The method of treatment of  claim 46 , wherein said disease is a disease selected from the group consisting of myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), chronic myeloid leukemia (CML) and pro-myelocytic leukemia (PML). 
     
     
         57 . A method of determining whether a biological sample contains a myelogenous cancer cell, comprising:
 (a) contacting said biological sample with a diagnostic reagent of  claim 29  or  30 , and   (b) detecting the distribution of said reagent within said sample.   
     
     
         58 . The method of diagnosis of  claim 57 , wherein said cancer is a cancer selected from the group consisting of acute myeloid leukemia (AML), chronic myeloid leukemia (CML) and pro-myelocytic leukemia (PML). 
     
     
         59 . An improved antibody or fragment thereof that specifically binds to CD33, said improved antibody or antibody fragment prepared by:
 (a) providing a DNA that encodes an antibody or epitope-binding fragment thereof comprising at least one of SEQ ID NO:7 and SEQ ID NO:8,   (b) introducing at least one nucleotide mutation, deletion, insertion or addition into said DNA such that the amino acid sequence of said antibody or epitope-binding fragment encoded by said DNA is changed;   (c) expressing said antibody or epitope-binding fragment;   (d) screening said expressed antibody or epitope-binding fragment for said improvement, thereby preparing an improved antibody or epitope-binding fragment.   
     
     
         60 . An improved antibody or epitope-binding fragment thereof that specifically binds to CD33, said improved antibody or antibody fragment prepared by:
 (a) providing a DNA that encodes an antibody or epitope-binding fragment thereof comprising at least one of SEQ ID NO:9 and SEQ ID NO:10,   (b) introducing at least one nucleotide mutation, deletion, insertion or addition into said DNA such that the amino acid sequence of said antibody or epitope-binding fragment encoded by said DNA is changed;   (c) expressing said antibody or epitope-binding fragment;   (d) screening said expressed antibody or epitope-binding fragment for said improvement, thereby preparing an improved antibody or epitope-binding fragment.   
     
     
         61 . The improved antibody or antibody fragment of  claim 59  or  60 , wherein said improvement is an increased affinity for CD33. 
     
     
         62 . The improved antibody or antibody fragment of  claim 59  or  60 , wherein said at least one nucleotide mutation, deletion, insertion or addition is made by a method selected from the group consisting of oligonucleotide-mediated site-directed mutagenesis, cassette mutagenesis, error-prone PCR, DNA shuffling and use of mutator-strains of  E. coli.    
     
     
         63 . An isolated polynucleotide encoding the antibody or epitope-binding fragment thereof of  claim 1  or  2 . 
     
     
         64 . An isolated polynucleotide encoding a light or heavy chain of the antibody or epitope-binding fragment thereof of  claim 1  or  2 . 
     
     
         65 . A recombinant vector comprising the polynucleotide of  claim 63 . 
     
     
         66 . A recombinant vector comprising the polynucleotide of  claim 64 . 
     
     
         67 . A host cell transformed with the recombinant vector of  claim 65 . 
     
     
         68 . A host cell transformed with the recombinant vector of  claim 66 . 
     
     
         69 . A method for producing an antibody or epitope-binding fragment thereof having the ability to bind CD33, said method comprising (a) culturing a host cell as claimed in  claim 67  under conditions such that said host cell expresses the antibody or epitope-binding fragment, and (b) collecting the antibody or epitope-binding fragment so expressed. 
     
     
         70 . A method for producing an antibody or epitope-binding fragment thereof having the ability to bind CD33, said method comprising (a) culturing a host cell as claimed in  claim 68  under conditions such that said host cell expresses the antibody or epitope-binding fragment, and (b) collecting the antibody or epitope-binding fragment so expressed. 
     
     
         71 . A method for obtaining CD33 from a biological material, said method comprising:
 (a) contacting a biological material with the antibody or epitope-binding fragment thereof of  claim 1  or  2 ,   (b) permitting the antibody or epitope-binding fragment of  claim 1  or  2  to bind to CD33 in said biological material, and   (c) isolating the antibody or epitope-binding fragment bound to CD33 from the biological material, thereby obtaining CD33 from a biological material.