Nanoparticle compositions and methods for immunotherapy
The present invention provides compositions and methods for immunotherapy, which include shelf-stable pharmaceutical compositions for inducing antigen-specific T cells. Such compositions are employed as components of an artificial antigen presenting cell (aAPC), to provide a patient with complexes for presentation of an antigen (e.g., a tumor antigen) and/or a T cell co-stimulatory molecule.
1. An artificial antigen presenting cell (aAPC) comprising a polymeric nanoparticle having attached polypeptide ligands suitable for activating or inhibiting antigen-specific T cells, the nanoparticle having a size in the range of 20 to 200 nm, the nanoparticle comprising:
a poly(lactic-co-glycolic acid) (PLGA) or polylactic acid (PLA) polymer core (PLGA/PLA), and
a hydrophilic shell formed of polyethylene glycol (PEG),
wherein the polymeric nanoparticle comprises PLGA/PLA-PEG and PLGA/PLA-methoxyPEG (mPEG) block co-polymers with the PLGA/PLA portion having molecular weights of from about 10 kDa to about 30 kDa and the PEG portion having molecular weight(s) of from about 2 kDa to about 10 kDa,
wherein a portion of the PLGA/PLA-PEG polymers have a terminal attachment of a polypeptide ligand, and
wherein the polypeptide ligands comprise HLA ligands presenting one or more peptide antigens.
2. The aAPC of claim 1 , wherein the PEG portions of the block co-polymers have a molecular weight of from about 3000 and/or about 5000 Da.
3. The aAPC of claim 1 , wherein the PLGA/PLA portions of the block co-polymers have a molecular weight of about 20,000 Da.
4. The aAPC of claim 1 , wherein the polymer core is based on a 20:1 to 1:20 Lactic Acid:Glycolic Acid ratio.
5. The aAPC of claim 1 , wherein the polymer core comprises PLA.
6. The aAPC of claim 1 , wherein the polypeptide ligands are attached to PEG through primary amines on the polypeptides.
7. The aAPC of claim 1 , wherein the polypeptide ligands are attached to PEG maleimide functional groups through an unpaired cysteine side chain.
8. The aAPC of claim 1 , wherein the aAPC has a surface charge of about 0 to −20 mV.
9. The aAPC of claim 1 , wherein the aAPC has a surface charge of from about −5 to about −10 mV.
10. The aAPC of claim 1 , having less than about 500 polypeptide ligands per particle.
11. The aAPC of claim 1 , having less than about 300 polypeptide ligands per particle.
12. The aAPC of claim 1 , wherein the polypeptide ligands further comprise a co-stimulatory or inhibitory ligand.
13. The aAPC of claim 12 , wherein the polypeptide ligands comprise an anti-CD28 agonistic antibody or an antigen-binding portion thereof.
14. The aAPC of claim 12 , wherein the polypeptide ligands comprise an antibody that binds to Fas.
15. The aAPC of claim 1 , wherein the HLA ligands are dimeric.
16. The aAPC of claim 15 , wherein the HLA polypeptide ligands are fused with co-stimulatory or inhibitory ligands.
17. The aAPC of claim 15 , wherein the HLA comprises a fusion with immunoglobulin sequences.
18. The aAPC of claim 17 , wherein the aAPC comprises an anti-CD28 antibody ligand, and each of the HLA immunoglobulin fusion polypeptide ligands and the anti-CD28 antibody ligands have an IgG4 constant region with mutations at codons S241 and L248, and an unpaired cysteine at codon 473.
19. The aAPC of claim 17 , wherein the HLA ligands comprise an extracellular domain of HLA fused to an immunoglobulin hinge region, without immunoglobulin variable domain sequences.