Compositions and methods for immunotherapy

Abstract

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.

What is claimed is: 
     
         1 . A pharmaceutical composition comprising:
 polymeric PLGA-PEG particles having a size in the range of about 100 to 200 nm, a surface charge of about 0 to −20 mV, and from about 100 to 1500 protein ligands per particle, the protein ligands optionally coupled through a sulfhydryl-mateimide chemistry;   a population of anti-CD28 antibody ligands;   a population of HLA ligands;   one or more antigenic peptides for presentation to T cells; and   a pharmaceutically acceptable carrier for intravenous, intra-arterial, subcutaneous, intradermal, intralymphatic, or intra-tumoral administration.   
     
     
         2 . The pharmaceutical composition of  claim 1 , wherein the particles are substantially spherical or about spherical. 
     
     
         3 . The pharmaceutical composition of  claim 1  or  2 , wherein the PLGA is a polymer of about 50% lactic acid (LA) and 50% glycolic acid (GA). 
     
     
         4 . The pharmaceutical composition of any one of  claims 1  to  3 , wherein the PLGA polymer has a molecular weight of from about 25K to about 35K, and the PEG has a molecular weight of from about 3K to about 10K. 
     
     
         5 . The pharmaceutical composition of any one of  claims 1  to  4 , having from 400 to 1000 ligands per particle. 
     
     
         6 . The pharmaceutical composition of any one of  claims 1  to  5 , wherein the anti-CD28 antibody ligands comprise a human IGHV4-59 germline framework optionally having from 5 to 15 murine framework residues, and a IGKV4-01 germline framework optionally having from 3 to 15 murine framework residues. 
     
     
         7 . The pharmaceutical composition of any one of  claims 1  to  5 , Wherein the anti-CD28 ligand is an antigen-binding antibody fragment. 
     
     
         8 . The pharmaceutical composition of any one of  claims 1  to  7 , wherein the anti-CD28 ligand comprises a scFv. 
     
     
         9 . The pharmaceutical composition of any one of  claims 1  to  8 , wherein the HLA ligand is dimeric. 
     
     
         10 . The pharmaceutical composition of  claim 9 , wherein the HLA is HLA-A*02:01. 
     
     
         11 . The pharmaceutical composition of  claim 9  or  10 , wherein the HLA comprises an immunoglobulin fusion. 
     
     
         12 . The pharmaceutical composition of  claim 11 , wherein the anti-CD28 antibody ligand and the HLA-A*02:01 immunoglobulin fusion have an IgG4 constant region with mutations at S241 and L248, and a Cysteine at codon 473. 
     
     
         13 . The pharmaceutical composition of  claim 12 , wherein the anti-CD28 antibody ligands are conjugated to the particles through the Cysteine at codon 473 of the immunoglobulin heavy chain. 
     
     
         14 . The pharmaceutical composition of  claim 12  or  13 , wherein the HLA ligands are conjugated to the particles through the Cysteine at codon 473 of the immunoglobulin heavy chain. 
     
     
         15 . The pharmaceutical composition of  claim 1 , wherein the HLA ligands comprise an extracellular domain of HLA, and are HLA monomers linked to the particle through a peptide or chemical linker attached to the C-terminus of the HLA extracellular domain, and wherein the HLA is optionally HLA-A2. 
     
     
         16 . The pharmaceutical composition of  claim 1 , wherein the HLA ligands comprise an extracellular domain of HLA, and are HLA monomers linked to both a scFv of an anti-CD28 ligand and the particle through one or more peptide or chemical linkers, and wherein the HLA is optionally HLA-A2. 
     
     
         17 . The pharmaceutical composition of any one of  claims 1  to  16 , wherein the composition is lyophilized. 
     
     
         18 . An anti-CD28 antibody or portion thereof, comprising a human IGHV4 heavy chain amino acid sequence optionally with from one to fifteen amino acid modifications, wherein at least one complementarity determining region (CDR) is based on a murine antibody amino acid sequence. 
     
     
         19 . The antibody of  claim 18 , wherein the CDRs are based on mouse 9.3 mAb. 
     
     
         20 . The antibody of  claim 18  or  19 , wherein the heavy chain is a variant of a human IGHV4-59 germline framework (FW). 
     
     
         21 . The antibody of any one of  claims 18  to  20 , wherein the antibody comprises a light chain and the light chain comprises a variant of a human IGKV4-01 FW. 
     
     
         22 . The antibody of any one of  claims 18  to  21 , wherein the heavy chain comprises one or more of murine Fw residues at positions selected from 1, 3, 6, 37, 48, 67, 71, 73, 76, 78, 82, 82a, and 82c. 
     
     
         23 . The antibody of  claim 22 , wherein the light chain comprises one or more murine Fw residues at positions selected from 3, 4, 49, 70, 85, 87, and 80. 
     
     
         24 . The antibody of any one of  claims 18  to  23 , wherein the antibody comprises a constant region or portion thereof, and the constant region is optionally a variant of human IgG4. 
     
     
         25 . The antibody of  claim 24 , wherein the constant region comprises one or more hinge stabilizing mutations. 
     
     
         26 . The antibody of  claim 25 , wherein the hinge stabilizing mutation comprises one or more mutations introduced in the CH chain. 
     
     
         27 . The antibody of  claim 26 , wherein the one or more mutations introduced in the CH chain includes S241P. 
     
     
         28 . The antibody of any one of  claims 18  to  27 , wherein the antibody comprises a constant region and the constant region comprises one or more mutations suitable for chemical coupling the antibody to a solid support. 
     
     
         29 . The antibody of  claim 28 , wherein the one or more mutations suitable for coupling create an unpaired cysteine. 
     
     
         30 . The antibody of  claim 29 , wherein the unpaired cysteine is S473C. 
     
     
         31 . The antibody of  claim 29  or  30 , wherein the unpaired cysteine provides for a covalent attachment to a solid support. 
     
     
         32 . The antibody of any one of  claims 18  to  31 , wherein the solid support is a bead or particle, optionally with functional groups, which are optionally esters, carboxylic acids, maleimides, alcohols, available for covalent bonding. 
     
     
         33 . The antibody of  claim 32 , wherein the bead or particle comprises a polymer selected from one or more of cyclodextrin-containing polymers, cationic cyclodextrin-containing polymers, poly(D,L-lactic acid-co-glycolic acid) (PLGA), poly(caprolactone) (PCL), ethylene vinyl acetate polymer (EVA), poly(lactic acid) (PLA), poly(L-lactic acid) (PLLA), poly(glycolic acid) (PGA), poly(L-lactic acid-co-glycolic acid) (PLLGA), poly(D,L-lactide), (PDLA), poly(L-lactide) (PLLA), PLGA-b-poly(ethylene glycol)-PLGA (PLGA-bPEG-PLGA), PLLA-bPEG-PLLA, PLGA-PEG-maleimide (PLGA-PEG-maI), poly(D,L-lactide-co-caprolactone), poly(D,L-lactide-co-caprotactone-co-glycolide), poly(D,L-lactide-co-PEO-co-D,L-lactide), poly(D,L-lactide-co-PPO-co-D,L-lactide), polyalkyl cyanoacralate, polyurethane, poly-L-lysine (PLL), hydroxypropyl methacrylate (HPMA), polyethyleneglycol, poly-L-glutamic acid, poly(hydroxy acids), polyanhydrides, polyorthoesters, poly(ester amides), polyamides, poly(ester ethers), polycarbonates, polyalkylenes such as polyethylene and polypropylene, polyalkylene glycols such as poly(ethylene glycol) (PEG), polyalkylene oxides (PEO), polyalkylene terephthalates such as poly(ethylene terephthalate), polyvinyl alcohols (PVA), polyvinyl ethers, polyvinyl esters such as poly(vinyl acetate), polyvinyl halides such as poly(vinyl chloride) (PVC), polyvinylpyrrolidone, polysiloxanes, polystyrene (PS), polyurethanes, derivatized celluloses such as alkyl celluloses, hydroxyalkyl celluloses, cellulose ethers, cellulose esters, nitro celluloses, hydroxypropylcellulose, carboxymethylcellulose, polymers of acrylic acids, such as poly(methyl(meth)acrylate) (PMMA), poly(ethyl(meth)acrylate), poly(butyl(meth)acrylate), poly(isobutyl(meth)acrylate), poly(hexyl(meth)acrylate), poly(isodecyl(meth)acrylate), poly(lauryl(meth)acrylate), poly(phenyl(meth)acrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), poly(octadecyl acrylate) (polyacrylic acids), and copolymers and mixtures thereof, polydioxanone and its copolymers, polyhydroxyalkanoates, polypropylene fumarate), polyoxymethylene, poloxamers, poly(ortho)esters, poly(butyric acid), poly(valeric acid), poly(lactide-co-caprotactone), trimethylene carbonate, polyvinylpyrrolidone, polyorthoesters, polyphosphazenes, and polyphosphoesters, and blends and/or block copolymers of two or more such polymers. 
     
     
         34 . The antibody of any one of  claims 18  to  33 , comprising a constant region, wherein the constant region comprises one or more mutations to reduce Fc gamma receptor binding. 
     
     
         35 . The antibody of  claim 34 , wherein the one or more mutations to reduce gamma receptor binding comprise one or more mutations introduced in the CH chain. 
     
     
         36 . The antibody of  claim 35 , wherein the one or more mutations introduced in the CH chain includes L248E. 
     
     
         37 . The antibody of any one of  claims 18  to  36 , wherein the antibody is conjugated to a solid support with molecular complexes presenting antigen for recognition by T cells. 
     
     
         38 . An antigen presenting complex comprising a humanized immunoglobulin heavy chain sequence fused to an HLA amino acid sequence, wherein the complex optionally does not contain an immunoglobulin light chain sequence. 
     
     
         39 . The antigen presenting complex of  claim 36 , wherein the HLA-Ig fusion is dimeric. 
     
     
         40 . The antigen presenting complex of  claim 38  or  39 , wherein the immunoglobulin sequence comprises an IGV4 sequence. 
     
     
         41 . The antigen presenting complex of any one of  claims 38  to  40 , wherein the HLA amino acid sequence is HLA-A*02:01 (IMGT Accession No. HLA00005) or a derivative thereof. 
     
     
         42 . The antigen presenting complex of any one of  claims 38  to  41 , further comprising a linker between the HLA and immunoglobulin sequences. 
     
     
         43 . The antigen presenting complex of any one of  claims 38  to  42 , further comprising an antigenic peptide bound to the HLA for presentation to T cells. 
     
     
         44 . The antigen presenting complex of  claim 43 , wherein the antigenic peptide is one or more of tyrosinase, MAGE-A1, MAGE-A3, gp100, A/Mart-1, gp75/brown, BAGE, NY-ESO-1 , and S-100. 
     
     
         45 . The antigen presenting complex of any one of  claims 38  to  44 , wherein the polypeptide is attached to solid support, which is optionally a bead or particle. 
     
     
         46 . The antigen presenting complex of  claim 45 , wherein the head or particle comprises a polymer selected from one or more of cyclodextrin-containing polymers, cationic cyclodextrin-containing polymers, poly(D,L-lactic acid-co-glycolic acid) (PLGA), poly(caprotactone) (PCL), ethylene vinyl acetate polymer (EVA), poly(lactic acid) (PLA), poly(L-lactic acid) (PLLA), PLGA-b-poly(ethylene glycol)-PLGA (PLGA-bPEG-PLGA), PLLA-bPEG-PLLA, PLGA-PEG-maleimide (PLGA-PEG-mal), poly(glycolic acid) (PGA), poly(L-lactic acid-co-glycolic acid) (PLLGA), poly(D,L-lactide) (PDLA), poly(L-lactide) (PLLA), poly(D,L-lactide-co-caprolactone), poly(D,L-lactide-co-caprolactone-co-glycolide), poly(D,L-lactide-co-PEO-co-D,L-lactide) poly(D,L-lactide-co-PPO-co-D,L-lactide), polyalkyl cyanoacralate, polyurethane, poly-L-lysine (PLL), hydroxypropyl methacrylate (HPMA), polyethyleneglycol, poly-L-glutamic acid, poly(hydroxy acids), polyanhydrides, polyorthoesters, poly(ester amides), polyamides, poly(ester ethers), polycarbonates, polyalkylenes such as polyethylene and polypropylene, polyalkylene glycols such as poly(ethylene glycol) (PEG), polyalkylene oxides (PEO), polyalkylene terephthalates such as poly(ethylene terephthalate), polyvinyl alcohols (PVA), polyvinyl ethers, polyvinyl esters such as poly(vinyl acetate), polyvinyl halides such as poly(vinyl chloride) (PVC), polyvinylpyrrolidone, polysiloxanes, polystyrene (PS), polyurethanes, derivatized celluloses such as alkyl celluloses, hydroxyalkyl celluloses, cellulose ethers, cellulose esters, nitro celluloses, hydroxypropylcellulose, carboxymethylcellulose, polymers of acrylic acids, such as poly(methyl(meth)acrylate) (PMMA), poly(ethyl(meth)acrylate), poly(butyl(meth)acrylate), poly(isobutyl(meth)acrylate), poly(hexyl(meth)acrylate), poly(isodecyl(meth)acrylate), poly(lauryl(meth)acrylate), poly(phenyl(meth)acrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), poly(octadecyl acrylate) (polyacrylic acids), and copolymers and mixtures thereof, polydioxanone and its copolymers, polyhydroxyalkanoates, polypropylene fumarate), polyoxymethylene, poloxamers, poly(ortho)esters, poly(butyric acid), poly(valeric acid), poly(lactide-co-caprolactone), trimethylene carbonate, polyvinylpyrrolidone, polyorthoesters, polyphosphazenes, and polyphosphoesters, and blends and/or block copolymers of two or more such polymers. 
     
     
         47 . A pharmaceutical composition comprising a polymeric bead or particle, an antibody according to any one of  claims 18  to  37 , and/or an antigen presenting complex of any one of  claims 38  to  46 . 
     
     
         48 . The pharmaceutical composition of  claim 47 , comprising polymeric PLGA-PEG particles, which arc optionally PLGA-PEG-maleimide particles, having a size in the range of about 100 to 200 nm. 
     
     
         49 . The pharmaceutical composition of  claim 47  or  48 , wherein the particles have a surface charge of about −0 to −15 mV, and optionally a surface charge of about −5 to about −15 mV. 
     
     
         50 . The pharmaceutical composition of any one of  claims 47  to  49 , comprising about 100 to 1500 protein ligands per particle, the protein ligands each coupled through a sulfhydryl-maleimide chemistry. 
     
     
         51 . The pharmaceutical composition of any one of  claims 47  to  50 , further comprising a pharmaceutically acceptable carrier for intravenous, intra-arterial, subcutaneous, intradermal, intralymphatic, or intra-tumoral administration. 
     
     
         52 . The pharmaceutical composition of any one of  claims 47  to  51 , wherein the particles are substantially spherical or about spherical. 
     
     
         53 . The pharmaceutical composition of any one of  claims 47  to  52 , wherein the PLGA is a polymer of about 50% lactic acid (LA) and 50% glycolic acid (GA). 
     
     
         54 . The pharmaceutical composition of any one of  claims 47  to  53 , wherein the PLGA polymer has a molecular weight of from about 25K to about 35K, and the PEG has a molecular weight of from about 3K to about 10K. 
     
     
         55 . The pharmaceutical composition of any one of  claims 47  to  54 , having from 400 to 1000 ligands per particle. 
     
     
         56 . The pharmaceutical composition of any one of  claims 47  to  55 , wherein the anti-CD28 ligand is an antigen-binding antibody fragment. 
     
     
         57 . The pharmaceutical composition of any one of  claims 47  to  56 , wherein the anti-CD28 ligand comprises a scFv. 
     
     
         58 . The pharmaceutical composition of any one of  claims 47  to  57 , Wherein the HLA ligand is dimeric. 
     
     
         59 . The pharmaceutical composition of  claim 58 , wherein the HLA is HLA-A*02:01. 
     
     
         60 . The pharmaceutical composition of  claim 58  or  59 , wherein the HLA comprises an immunoglobulin heavy chain fusion, and optionally does not comprise an immunoglobulin light chain. 
     
     
         61 . The pharmaceutical composition of any one of  claims 47  to  60 , further comprising an antigenic peptide for presentation to T cells. 
     
     
         62 . The pharmaceutical composition of  claim 61 , wherein the pharmaceutical composition is co-formulated with an antigenic peptide for presentation to T cells. 
     
     
         63 . The pharmaceutical composition of  claim 61  or  62 , wherein the antigenic peptide is one or more of tyrosinase, MAGE-A1, MAGE-A3, gp100, Melan A/Mart-1, gp75/brown, BAGE, NY-ESO-1, and S-100. 
     
     
         64 . The pharmaceutical composition of army one of  claims 47  to  63 , wherein the pharmaceutical composition is shelf stable. 
     
     
         65 . The pharmaceutical composition of  claim 64 , wherein the pharmaceutical composition is lyophilized. 
     
     
         66 . The pharmaceutical composition of any one of  claims 47  to  65 , wherein the composition is formulated for parenteral delivery. 
     
     
         67 . A method for inducing the formation of antigen-specific cytotoxic T cells, comprising administering a composition of any one of  claims 47  to  66  to a patient. 
     
     
         68 . The method of  claim 67 , wherein the patient is a cancer patient. 
     
     
         69 . The method of  claim 67  or  68 , wherein the patient is undergoing or has undergone therapy with one or more checkpoint inhibitors.