Gamma polyglutamated pralatrexate and uses thereof
The disclosure relates generally to gamma polyglutamated pralatrexate compositions, including delivery vehicles such as liposomes containing the gamma polyglutamated pralatrexate, and methods of making and using the gamma polyglutamated pralatrexate compositions to treat hyperproliferative disorders (e.g., cancer) and disorders of the immune system (e.g., inflammation and autoimmune diseases such as rheumatoid arthritis).
1. A liposomal composition comprising a gamma polyglutamated pralatrexate encapsulated by a liposome, wherein the polyglutamated pralatrexate has 4, 5, 2-10, 4-6, or more than 5, glutamyl groups having gamma carboxyl group linkages, wherein the liposome is pegylated and does not contain a targeting moiety having specific affinity for a surface antigen on a target cell,
wherein the liposome does not contain a cell penetrating peptide and does not contain a mitochondria penetrating agent,
wherein the liposome has a zeta potential that is less than or equal to zero, between 0 and −150 mV, or between −30 and −50 mV, and
wherein the liposome is capable of delivering the gamma polyglutamated pralatrexate directly into a cell.
2. The liposomal composition of claim 1 , wherein the liposome further comprises one or more nonpolyglutamated polyglutamatable antifolate or non-polyglutamatable antifolate.
3. The liposomal composition of claim 1 , wherein the gamma polyglutamated pralatrexate has 4, 5, 2-10, or 4-6 glutamyl groups.
4. The liposomal composition of claim 1 , wherein the gamma polyglutamated pralatrexate comprises gamma tetraglutamated pralatrexate.
5. The liposomal composition of claim 1 , wherein the gamma polyglutamated pralatrexate comprises gamma pentaglutamted pralatrexate.
6. The liposomal composition of claim 1 , wherein the gamma polyglutamated pralatrexate comprises gamma hexaglutamted pralatrexate.
7. The liposomal composition of claim 1 , wherein the liposome has a diameter in the range of 20 nm to 500 nm, 20 nm to 200 nm, or 80 nm to 120 nm.
8. The liposomal composition of claim 1 , wherein the liposome is formed from liposomal components comprising:
at least one of an anionic lipid and a neutral lipid;
at least one selected from: DSPE; DSPE-PEG; DSPE-PEG-maleimide; HSPC; HSPC-PEG;
cholesterol; cholesterol-PEG; and cholesterol-maleimide; or
at least one selected from: DSPE; DSPE-PEG; DSPE-PEG-FITC; DSPE-PEG-maleimide;
cholesterol; and HSPC.
9. The liposomal composition of claim 8 , wherein one or more liposomal components further comprises at least one steric stabilizer selected from: 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; oligoglycerol, copolymer containing polyethylene glycol and polypropylene oxide, Poloxamer 188, and polyvinyl alcohol.
10. The liposomal composition of claim 1 , wherein the liposome is in an aqueous pharmaceutically acceptable carrier comprising:
a tonicity agent at a concentration of greater than 1%;
1% to 50% trehalose;
1% to 50% dextrose;
5% dextrose suspended in an HEPES buffered solution; or
a total concentration of sodium acetate and calcium acetate of between 50 mM to 500 mM.
11. The liposomal composition of claim 1 , wherein the interior space of the liposome has a pH of 5-8 or a pH of 6-7, or any range therein between.
12. The liposomal composition of claim 1 , wherein the liposome comprises between 10 and 500,000, between 10 and 200,000, or between 10 to 100,000 molecules of the gamma polyglutamated pralatrexate, or any range therein between.
13. The liposomal composition of claim 1 , which further comprises at least one cryoprotectant selected from mannitol, trehalose, sorbitol, and sucrose.
14. The liposomal composition of claim 1 , which further comprises carboplatin or pembrolizumab.
15. The liposomal composition of claim 2 , wherein the nonpolyglutamated polyglutamatable antifolate is selected from: methotrexate (MTX), pemetrexed (PMX), lometrexol (LMX), raltitrexed (RTX), pralatrexate, AG2034, GW1843, aminopterin, and LY309887; or the non-polyglutamatable antifolate is selected from: trimetrexate (TMQ), piritrexim (BW301U), and talotrexin (PT523), nolatrexed (AG337), plevitrexed, and BGC 945 (ONX 0801).
16. The liposomal composition of claim 1 , wherein the polyglutamated pralatrexate has 2-10 glutamyl groups having gamma carboxyl group linkages.
17. The liposomal composition of claim 1 , wherein the polyglutamated pralatrexate has 4-6 glutamyl groups having gamma carboxyl group linkages.
18. The liposomal composition of claim 1 , wherein the liposome encapsulates between 10-100,000 molecules of polyglutamated pralatrexate having 2-10 glutamyl groups.
19. A pharmaceutical composition comprising the liposomal composition of claim 1 .
20. A method of killing a hyperproliferative cell that comprises contacting a hyperproliferative cell with the liposomal composition of claim 1 .
21. The method of claim 20 , wherein the hyperproliferative cell is a cancer cell, a mammalian cell, and/or a human cell.
22. A method for treating cancer that comprises administering an effective amount of the liposomal composition of claim 1 to a subject having cancer.
23. A method of preparing a gamma polyglutamated pralatrexate composition comprising the liposomal composition of claim 1 , the method comprising: forming a mixture comprising: liposomal components and gamma polyglutamated pralatrexate in solution; homogenizing the mixture to form liposomes in the solution; and processing the mixture to form liposomes containing gamma polyglutamated pralatrexate.