LAG-3-binding molecules and methods of use thereof
The present invention is directed to the anti-LAG-3 antibodies: LAG-3 mAb 1, LAG-3 mAb 2, LAG-3 mAb 4, LAG-3 mAb 5, and LAG-3 mAb 6, and to humanized and chimeric versions of such antibodies. The invention additionally pertains to LAG-3-binding molecules that comprise LAG-3 binding fragments of such anti-LAG-3 antibodies, immunoconjugates, and to bispecific molecules, including diabodies, BiTEs, bispecific antibodies, etc., that comprise (i) such LAG-3-binding fragments, and (ii) a domain capable of binding an epitope of a molecule involved in regulating an immune check point present on the surface of an immune cell. The present invention also pertains to methods of detecting LAG-3, as well as methods of using molecules that bind LAG-3 for stimulating immune responses.
1. A method of stimulating a T-cell-mediated immune response in a subject in need thereof comprising administering to said subject a therapeutically effective amount of a Lymphocyte Activation Gene 3 (LAG-3)-binding molecule that is capable of binding both to human LAG-3 and to cynomolgus monkey LAG-3, wherein said LAG-3-binding molecule comprises a Variable Heavy Chain Domain and a Variable Light Chain Domain, wherein: said Variable Heavy Chain Domain comprises a CDR H 1 Domain, a CDR H 2 Domain and a CDR H 3 Domain, and said Variable Light Chain Domain comprises a CDR L 1 Domain, a CDR L 2 Domain, and a CDR L 3 Domain, wherein:
(A) (1) the CDR H 1 Domain, CDR H 2 Domain, and CDR H 3 Domain are the Heavy Chain CDRs of LAG-3 mAb 1, and respectively have the amino acid sequences: SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:10; and
(2) the CDR L 1 Domain, CDR L 2 Domain, and CDR L 3 Domain are the Light Chain CDRs of LAG-3 mAb 1, and respectively have the amino acid sequences: SEQ ID NO:13, SEQ ID NO:14, and SEQ ID NO:15;
or
(B) (1) the CDR H 1 Domain, CDR H 2 Domain, and CDR H 3 Domain are the Heavy Chain CDRs of hLAG-3 mAb 1, and respectively have the amino acid sequences: SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:10; and
(2) the CDR L 1 Domain, CDR L 2 Domain, and CDR L 3 Domain are the Light Chain CDRs of hLAG-3 mAb 1, and respectively have the amino acid sequences: SEQ ID NO:28, SEQ ID NO:14, and SEQ ID NO:15;
or
(C) (1) the CDR H 1 Domain, CDR H 2 Domain, and CDR H 3 Domain are the Heavy Chain CDRs of LAG-3 mAb 6 VH1, and respectively have the amino acid sequences: SEQ ID NO:71, SEQ ID NO:72, and SEQ ID NO:73; and
(2) the CDR L 1 Domain, CDR L 2 Domain, and CDR L 3 Domain are the Light Chain CDRs of LAG-3 mAb 6, and, respectively have the amino acid sequences: SEQ ID NO:76, SEQ ID NO:77, and SEQ ID NO:78;
or
(D) (1) the CDR H 1 Domain, CDR H 2 Domain, and CDR H 3 Domain are the Heavy Chain CDRs of LAG-3 mAb 6, and respectively have the amino acid sequences: SEQ ID NO:71, SEQ ID NO:72, and SEQ ID NO:73; and
(2) the CDR L 1 Domain, CDR L 2 Domain, and CDR L 3 Domain are the Light Chain CDRs of hLAG-3 mAb 6, and respectively have the amino acid sequences: SEQ ID NO:87, SEQ ID NO:77, and SEQ ID NO:78.
2. The method of claim 1 , wherein said molecule is an antibody.
3. The method of claim 2 , wherein said molecule is a chimeric antibody or a humanized antibody.
4. The method of claim 1 , wherein said molecule comprises:
(a) a Heavy Chain Variable Domain having the amino acid sequence of SEQ ID NO:16; and
(b) a Light Chain Variable Domain having the amino acid sequence of SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, or SEQ ID NO:26.
5. The method of claim 1 , wherein:
(1) the CDR H 1 Domain, CDR H 2 Domain, and CDR H 3 Domain are the Heavy Chain CDRs of LAG-3 mAb 1, and respectively comprise the amino acid sequences: SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:10; and
(2) the CDR L 1 Domain, CDR L 2 Domain, and CDR L 3 Domain are the Light Chain CDRs of LAG-3 mAb 1, and respectively comprise the amino acid sequences: SEQ ID NO:13, SEQ ID NO:14, and SEQ ID NO:15.
6. The method of claim 5 , wherein said Variable Heavy Chain Domain comprises the amino acid sequence of SEQ ID NO:16, and said Variable Light Chain Domain comprises the amino acid sequence of SEQ ID NO:24.
7. The method of claim 1 , wherein:
(1) the CDR H 1 Domain, CDR H 2 Domain, and CDR H 3 Domain are the Heavy Chain CDRs of LAG-3 mAb 1, and respectively comprise the amino acid sequences: SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:10; and
(2) the CDR L 1 Domain, CDR L 2 Domain, and CDR L 3 Domain are the Light Chain CDRs of LAG-3 mAb 1, and respectively comprise the amino acid sequences: SEQ ID NO:28, SEQ ID NO:14, and SEQ ID NO:15.
8. The method of claim 7 , wherein, said Variable Heavy Chain Domain comprises the amino acid sequence of SEQ ID NO:16, and said Variable Light Chain Domain comprises the amino acid sequence of SEQ ID NO:26.
9. The method of claim 1 , wherein:
(1) the CDR H 1 Domain, CDR H 2 Domain, and CDR H 3 Domain are the Heavy Chain CDRs of hLAG-3 mAb 6, and respectively comprise the amino acid sequences: SEQ ID NO:71, SEQ ID NO:72, and SEQ ID NO:73; and
(2) the CDR L 1 Domain, CDR L 2 Domain, and CDR L 3 Domain are the Light Chain CDRs of hLAG-3 mAb 6, and respectively comprise the amino acid sequences: SEQ ID NO:87, SEQ ID NO:77, and SEQ ID NO:78.
10. The method of claim 9 , wherein said molecule comprises:
(a) a Heavy Chain Variable Domain comprising the amino acid sequence of SEQ ID NO: 79; and
(b) a Light Chain Variable Domain comprising the amino acid sequence of SEQ ID NO:83.
11. The method of claim 1 , wherein said molecule comprises an Fc Region.
12. The method of claim 11 , wherein said Fc Region is a variant Fc Region that comprises:
(a) one or more amino acid modifications that reduce(s) the affinity of said variant Fc Region for an FcγR, wherein said modifications comprise the substitutions:
(1) L234A;
(2) L235A; or
(3) L234A and L235A; and/or
(b) one or more amino acid modifications that enhance(s) the serum half-life of said variant Fc Region, wherein said modifications comprise the substitutions:
(1) M252Y and S254T;
(2) M252Y and T256E;
(3) M252Y, S254T and T256E; or
(4) K288D and H435K;
wherein said numbering is that of the EU index according to Kabat.
13. The method of claim 6 , wherein said molecule comprises an Fc Region.
14. The method of claim 13 , wherein said Fc Region is a variant Fc Region that comprises:
(a) one or more amino acid modifications that reduce(s) the affinity of said variant Fc Region for an FcγR, wherein said modifications comprise the substitutions:
(1) L234A;
(2) L235A; or
(3) L234A and L235A; and/or
(b) one or more amino acid modifications that enhance(s) the serum half-life of said variant Fc Region, wherein said modifications comprise the substitutions:
(1) M252Y and S254T;
(2) M252Y and T256E;
(3) M252Y, S254T and T256E; or
(4) K288D and H435K;
wherein said numbering is that of the EU index according to Kabat.
15. The method of claim 8 , wherein said molecule comprises an Fc Region.
16. The method of claim 15 , wherein said Fc Region is a variant Fc Region that comprises:
(a) one or more amino acid modifications that reduce(s) the affinity of said variant Fc Region for an FcγR, wherein said modifications comprise the substitutions:
(1) L234A;
(2) L235A; or
(3) L234A and L235A; and/or
(b) one or more amino acid modifications that enhance(s) the serum half-life of said variant Fc Region, wherein said modifications comprise the substitutions:
(1) M252Y and S254T;
(2) M252Y and T256E;
(3) M252Y, S254T and T256E; or
(4) K288D and H435K;
wherein said numbering is that of the EU index according to Kabat.
17. The method of claim 10 , wherein said molecule comprises an Fc Region.
18. The method of claim 17 , wherein said Fc Region is a variant Fc Region that comprises:
(a) one or more amino acid modifications that reduce(s) the affinity of said variant Fc Region for an FcγR, wherein said modifications comprise the substitutions:
(1) L234A;
(2) L235A; or
(3) L234A and L235A; and/or
(b) one or more amino acid modifications that enhance(s) the serum half-life of said variant Fc Region, wherein said modifications comprise the substitutions:
(1) M252Y and S254T;
(2) M252Y and T256E;
(3) M252Y, S254T and T256E; or
(4) K288D and H435K;
wherein said numbering is that of the EU index according to Kabat.