Alpha and gamma-D polyglutamated pemetrexed antifolates and uses thereof
The disclosure relates generally to polyglutamated antifolates, formulations containing liposomes filled with alpha or D-gamma polyglutamated antifolates, methods of making the polyglutamated antifolates and liposome containing formulations, and methods of using polyglutamated antifolates and liposome containing formulations to treat hyerproliferative disorders (e.g., cancer) and disorders of the immune system (e.g., an autoimmune disease such as rheumatoid arthritis).
1. A liposomal composition comprising polyglutamated pemetrexed (PMX) encapsulated by a pegylated liposome, wherein the polyglutamated PMX comprises at least 1 D-glutamate residue.
2. The liposomal composition of claim 1 , wherein the polyglutamated PMX comprises 3, 4, 5, 6, 7, 8, 9, or 2-10 D-glutamate residues.
3. The liposomal composition of claim 1 , wherein the polyglutamated PMX comprises 4, 5, 5-10, or more than 5, glutamate residues.
4. The liposomal composition of claim 1 , wherein the polyglutamated PMX comprises at least 3 glutamate residues linked by alpha carboxy group linkages.
5. The liposomal composition of claim 1 , wherein the polyglutamated PMX comprises at least 2 glutamate residues linked by gamma carboxy group linkages.
6. The liposomal composition of claim 1 , wherein the polyglutamated PMX comprises pentaglutamated PMX.
7. The liposomal composition of claim 1 , wherein the polyglutamated PMX comprises 4 D-gamma glutamate residues.
8. The liposomal composition of claim 1 , wherein the polyglutamated PMX comprises hexaglutamated PMX.
9. The liposomal composition of claim 1 , wherein the polyglutamated PMX comprises 5 D-gamma glutamate residues.
10. The liposomal composition of claim 1 , wherein the polyglutamated PMX comprises at least 1 D-glutamate residue and at least one L-glutamate residue.
11. The liposomal composition of claim 1 , wherein the liposome has a diameter in the range of 20 nm to 200 nm or 80 nm to 120 nm.
12. The liposomal composition of claim 1 , wherein the liposome is formed from liposomal components and wherein: the liposomal components comprise:
(a) at least one of an anionic lipid and a neutral lipid;
(b) at least one member selected from the group consisting of: DSPE; DSPE-PEG; DSPE-PEG-maleimide; HSPC; HSPC-PEG; cholesterol; cholesterol-PEG; and cholesterol-maleimide; or
(c) at least one selected from the group consisting of: DSPE; DSPE-PEG-FITC; DSPE-PEG-maleimide; cholesterol; and HSPC.
13. The liposomal composition of claim 12 , wherein one or more liposomal components further comprises at least one steric stabilizer selected from the group consisting of polyethylene glycol (PEG); poly-L-lysine (PLL); monosialoganglioside (GM1); poly(vinyl pyrrolidone) (PVP); poly(acrylamide) (PAA); poly(2-methyl-2-oxazoline); poly(2-ethyl-2-oxazoline); phosphatidyl polyglycerol; poly[N-(2-hydroxypropyl) methacrylamide]; amphiphilic poly-N-vinylpyrrolidones; L-amino-acid-based polymer; or polyvinyl alcohol.
14. The liposomal composition of claim 1 , wherein the PEG of the liposome has a number average molecular weight (Mn) of 200 to 5000 daltons.
15. The liposomal composition of claim 1 , wherein the liposome:
(a) is anionic or neutral,
(b) has a zeta potential that is less than or equal to zero,
(c) has a zeta potential that is between 0 to −150 mV, or
(d) has a zeta potential that is between −30 to −50 mV.
16. The liposomal composition of claim 1 , wherein the liposome is cationic.
17. The liposomal composition of claim 1 , wherein the liposome has an interior space and the interior space comprises the polyglutamated PMX and an aqueous pharmaceutically acceptable carrier, optionally wherein the aqueous pharmaceutically acceptable carrier (a) is trehalose, (b) comprises citrate buffer at a concentration of between 5 to 200 mM and a pH of between 2.8 to 6, or (c) comprises a total concentration of sodium acetate and calcium acetate of between 50 mM to 500 mM.
18. The liposomal composition of claim 1 , wherein the liposome comprises less than 200,000 molecules or between 10,000 to 100,000 molecules of polyglutamated PMX.
19. The liposomal composition of claim 1 , wherein the polyglutamated PMX comprises at least 2 D glutamate residues and the liposome has a diameter in the range of 20 nm to 200 and a zeta potential that is less than or equal to zero.
20. The liposomal composition of claim 19 , wherein the polyglutamated PMX comprises 3, 4, 5, 6, 7, 8, 9, or 2-10 D-glutamate residues.
21. The liposomal composition of claim 1 , which further comprises a targeting moiety attached to one or both of a PEG and the exterior of the liposome, optionally by a covalent bond, and wherein the targeting moiety has a specific affinity for a surface antigen on a target cell of interest.
22. The liposomal composition of claim 21 , wherein the targeting moiety is a polypeptide, or one or more selected from the group consisting of: an antibody, a humanized antibody, an antigen binding fragment of an antibody, a single chain antibody, a single-domain antibody, a bi-specific antibody, a synthetic antibody, a pegylated antibody, and a multimeric antibody.
23. The liposomal composition of claim 21 , wherein the targeting moiety binds the antigen with an equilibrium dissociation constant (Kd) in a range of 0.5×10 −10 to 10×10 −6 as determined using surface plasmon resonance analysis.
24. The liposomal composition of claim 21 , wherein the targeting moiety specifically binds one or more folate receptors selected from the group consisting of: folate receptor alpha (FR-α), folate receptor beta (FR-β), and folate receptor delta (FR-δ).
25. The liposomal composition of claim 1 , wherein each liposome comprises from 30 to 200 targeting moieties.
26. The liposomal composition of claim 1 , further comprising one or more of an immunostimulating agent, a detectable marker and a maleimide, wherein the immunostimulatory agent, the detectable marker or the maleimide is attached to said PEG or the exterior of the liposome.
27. The liposomal composition of claim 26 , wherein the immunostimulating agent is at least one selected from the group consisting of: a fluorescein; a fluorescein isothiocyanate (FITC); a dinitrophenol (DNP); a beta glucan; a beta-1,3-glucan; and a beta-1,6-glucan.
28. The liposomal composition of claim 1 , further comprising a hapten or at least one cryoprotectant selected from the group consisting of mannitol; trehalose; sorbitol; and sucrose.
29. The liposomal composition of claim 1 , which is in unit dosage form.
30. A pharmaceutical composition comprising the liposomal composition of claim 1 .