Targeted CD1d molecules
View Patent ↗The invention is directed to a compound comprising one or more CD1d complexes in association with an antibody specific for a cell surface marker. The CD1d complexes comprise a CD1d, a β2-microglobulin molecule, and may further comprise an antigen bound to the CD1d binding groove. The invention is further directed to methods of inhibiting or stimulating an immune response with the CD1d-antibody compounds, in particular anti-tumor and autoimmunity responses.
1. A compound comprising:
(a) a soluble CD1d complex comprising:
i) a CD1d molecule or antigen-binding fragment thereof;
ii) a β2-microglobulin molecule or fragment thereof; and
iii) a stimulatory glycolipid antigen bound in the antigen binding groove of the CD1d molecule or antigen-binding fragment thereof; and
(b) an antibody or an antigen-binding fragment thereof specific for a cell surface marker of a tumor cell;
wherein said CD1d complex is linked to said antibody or fragment thereof; and wherein said compound stimulates the activation of the cytolytic activity of NKT cells.
2. The compound of claim 1 , wherein said antigen is α-GalCer.
3. The compound of claim 1 , wherein said antigen is a modified α-GalCer that is the OCH analog thereof having a long-chain sphingosine base shortened from C14 to C5 and acyl chain from C26 to C24.
4. The compound of claim 1 , wherein said antigen-binding antibody fragment is a F(ab).
5. The compound of claim 1 , wherein said antigen-binding antibody fragment is a scFv.
6. The compound of claim 1 , wherein said antibody is a full-length antibody.
7. The compound of claim 1 , wherein said cell surface marker is selected from the group consisting of: CEA, Her2/neu, EGFR type I or type II, CD19, CD20, CD22, Muc-1, PSMA, and STEAP.
8. The compound of claim 1 , wherein said CD1d molecule or antigen-binding fragment thereof is attached to the heavy chain of said antibody.
9. The compound of claim 1 , wherein said CD1d molecule or antigen-binding fragment thereof is attached to the light chain of said antibody.
10. The compound of claim 1 , wherein said β2 microglobulin molecule is attached to the heavy chain of said antibody.
11. The compound of claim 1 , wherein said β2 microglobulin molecule is attached to the light chain of said antibody.
12. The compound of claim 1 , wherein the CD1d complex is linked in a fusion protein with the antibody or antigen-binding fragment thereof.
13. The compound of claim 7 , wherein said cell surface marker is Her2/neu.
14. The compound of claim 1 , wherein the CD1d molecule or antigen-binding fragment thereof of said CD1d complex comprises the extracellular portion of CD1d.
15. The compound of claim 14 , wherein said extracellular portion comprises amino acids 1-297 of the amino acid sequence of SEQ ID NO: 40.
16. The compound of claim 5 , wherein the variable light domain and the variable heavy domain of said scFv are linked by a peptide bridge.
17. The compound of claim 5 , wherein the variable light domain and the variable heavy domain of said scFv are linked by one or more disulfide bonds.
18. The compound of claim 12 , wherein the CD1d molecule of said CD1d complex is fused to said antibody or antigen-binding fragment thereof.
19. The compound of claim 18 , wherein the CD1d molecule of said CD1d complex is fused to the amino terminus of the antibody or antigen-binding fragment thereof.
20. The compound of claim 18 , wherein the CD1d molecule of said CD1d complex is fused to the carboxyl terminus of the antibody or antigen-binding fragment thereof.
21. The compound of claim 18 , wherein a short linker amino acid sequence consisting of at least 3 and not more than 30 amino acids is situated between the CD1d molecule of said CD1d complex and the antibody or antigen-binding fragment thereof.
22. The compound of claim 21 , wherein said short linker amino acid sequence comprises the sequence of SEQ ID NO: 1.
23. The compound of claim 21 , wherein said short linker amino acid sequence comprises the sequence of SEQ ID NO: 2.
24. The compound of claim 12 , wherein the β2-microglobulin molecule of said CD1d complex is fused to said antibody or antigen-binding fragment thereof.
25. The compound of claim 24 , wherein the β2-microglobulin molecule of said CD1d complex is fused to the amino terminus of the antibody or antigen-binding fragment thereof.
26. The compound of claim 24 , wherein the β2-microglobulin molecule of said CD1d complex is fused to the carboxyl terminus of the antibody or antigen-binding fragment thereof.
27. The compound of claim 24 , wherein a short linker amino acid sequence of from about 3 to about 30 amino acids is situated between the β2-microglobulin molecule of said CD1d complex and the antibody or antigen-binding fragment thereof.
28. The compound of claim 27 , wherein said short linker amino acid sequence comprises the sequence of SEQ ID NO: 1.
29. The compound of claim 27 , wherein said short linker amino acid sequence comprises the sequence of SEQ ID NO: 2.