Gamma polyglutamine oxoxide antimetabolites and their use

Abstract

To provide a gamma-polyglutamate oxidase folate antagonist composition that can overcome pharmacological challenges related to dose-limiting toxicity and therapeutic resistance associated with folate antagonist therapy. [Solution] A liposome composition (Lp-γPANTIFOL composition) containing a gamma polyglutamine oxidized folate antimetabolitic agent, wherein the gamma polyglutamine oxidized folate antimetabolitic agent contains 2 to 15 glutamyl groups, and the liposomes are pegylated.

Description

Background Art

[0001] The present disclosure generally relates to compositions of gamma-polyglutamylated folate antagonists, such as liposomes containing delivery carriers of gamma-polyglutamylated folate antagonist compositions, and methods of manufacturing and using the compositions for treating diseases including hyperproliferative diseases such as cancer, immune system disorders including inflammation and autoimmune diseases such as rheumatoid arthritis, and infectious diseases such as HIV, malaria, and schistosomiasis.

[0002] Folic acid is an essential cofactor that mediates the transfer of one-carbon units involved in biosynthesis and DNA repair, remethylation of homocysteine (Hcy), and methylation of DNA, proteins, and lipids. The only circulating form of folic acid in the blood is monoglutamic acid, and folic acid monoglutamate is the only form of folic acid that is transported across the cell membrane. Similarly, monoglutamic acid-type polyglutamylatable folate antagonists are also transported across the cell membrane. Once taken up into the cell, intracellular folic acid is converted to polyglutamic acid by the enzyme folylpolyglutamate synthetase (FPGS).

[0003] Folate antagonists are transported into cells by the reduced folate carrier (RFC) system and folate receptors (FR) α and β, as well as by the proton-coupled folate transporter (PCFT), which is most active at a lower pH than normal. RFC is the major transporter of folate antagonists at physiological pH and is widely expressed in normal and diseased cells. Thus, folate antagonist therapy often suffers from dose-limiting toxicity, a major obstacle in cancer chemotherapy. Once inside the cell, folate antagonists are polyglutamylated by FPGS, which can add up to six glutamyl groups during the binding of the L-gamma carboxyl group to the folate antagonist. L-gamma polyglutamylation of folate antagonists by FPGS serves at least two major therapeutic purposes: (1) it greatly enhances the affinity and inhibitory activity of folate antagonists against DHFR; and (2) it facilitates the accumulation of polyglutamylated folate antagonists, which, unlike folate antagonists (monoglutamate), are not readily transported out of cells by cellular efflux pumps.

[0004] Targeting folate metabolism and nucleotide biosynthesis is an established therapeutic strategy for cancer, but for folate antagonists, clinical efficacy is limited by the lack of tumor selectivity and the presence of new and acquired drug resistance. Folate antagonists often act during DNA and RNA synthesis and, as a result, have a major toxic effect on rapidly dividing cells such as malignant and myeloid cells. Myelosuppression is usually the dose-limiting toxicity of folate antagonist therapy and restricts the clinical application of folate antagonists.

[0005] Resistance to folate antagonist therapy is usually associated with one or more of the following: (a) increased activity of cellular efflux pumps, (b) decreased transport of folate antagonists into cells, (c) increased DHFR activity, (d) decreased activity of holylpolyglutamate synthase (FPGS), and (e) increased activity of gamma-glutamyl hydrolase (GGH), which cleaves the gamma-polyglutamate chain attached to folate and folate antagonists.

[0006] The problem with the long-term (>30-year) observation that higher levels of polyglutamates of various folate antimetabolites are far more potent than lower levels of glutamates has been that the scientific community has relied on intracellular FPGS-mediated mechanisms to convert low levels of glutamates to their higher-level forms. This invention provides a means for directly delivering higher-level polyglutamate forms of folate antimetabolites into cells, without relying on cellular mechanisms to achieve this goal.

[0007] The provided gamma-polyglutamine-oxidized folate antimetabolites offer strategies to overcome the pharmacological challenges associated with dose-limiting toxicity and therapeutic resistance related to folate antimetabolites therapy. In some embodiments, the provided methods deliver gamma-polyglutamine-oxidized folate antimetabolites to cancer cells while simultaneously (1) minimizing / reducing exposure to normal tissue cells, (2) optimizing / improving the cytotoxic effect of the folate antimetabolite-based agent on cancer cells, and (3) minimizing / reducing the effects of efflux pumps and other resistance mechanisms that limit the therapeutic efficacy of folate antimetabolites. [Overview of the project]

[0008] This disclosure generally relates to gamma polyglutamine oxidase antimetabolites (γPANTIFOL) compositions, and to methods for manufacturing and using compositions for treating diseases including hyperproliferative diseases such as cancer, inflammation and immune system disorders such as rheumatoid arthritis, cardiovascular diseases such as coronary artery disease, and infectious diseases such as HIV, malaria, and schistosomiasis.

[0009] In some embodiments, this disclosure provides the following: [1] A composition comprising a gamma-polyglutamine oxoxide antimetabolitic agent; [2] A composition according to item [1], wherein the folate antagonist is selected from pyritrexime, pralatrexate, AG2034, GW1843, and LY309887, or their stereoisomers; [3] A composition according to item [1], wherein the folate antagonist is selected from PMX, MTX, RTX, and LTX, or their stereoisomers; [4] A composition described in any of items [1] to [3], wherein the folate antagonist is selected from the following: LV (etoposide), L-leucovorin (L-5-formyltetrahydrofolate); 5-CH3-THF, 5-methyltetrahydrofolate; FA, folate; PteGlu, pteroylglutamate (FA); MTX, methotrexate; 2-dMTX, 2-desamino-MTX; 2-CH3-MTX, 2-desamino-2-methyl-MTX; AMT, aminopterin; 2-dAMT, 2-desamino-AMT; 2-CH3-AMT, 2-desamino-2-methyl-AMT; 10-EdAM, 10-ethyl-10-deazaaminopterin; PT523, N alpha-(4-amino-4-deoxypteroyl)-N Delta-(hemiphthaloyl)-L-ornithine;DDATHF (lometrexol), 5,10-dideaza-5,6,7,8-tetrahydrofolate;5-d(i)H4PteGlu, 5-deaza-5,6,7,8-tetrahydroisofolate;N9-CH3-5-d(i)H4PteGlu, N9-methyl-5-deaza-5,6,7,8-tetrahydroisofolate;5-dPteHCysA,N Alpha-(5-deazapteroyl)-L-homocysteic acid;5-dPteAPBA,N Alpha-(5-deazapteroyl)-DL-2-amino-4-phosphonobutyric acid;5-dPteOrn,N Alpha-(5-deazapteroyl)-L-ornithine;5-dH4PteHCysA,N Alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-L-homocysteic acid; 5-dH4PteAPBA,N Alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-DL-2-amino-4-phosphobutanoic acid; 5-dH4PteOro,N Alpha-(5-dideaza-5,6,7,8-tetrahydropteroyl)-L-ornithine;CB3717, N10-propargyl-5,8-dideazafolate;ICI-198,583, 2-desamino-2-methyl-N10-propargyl-5,8-dideazafolate;4-H-ICI-198,583, 4-deoxy-ICI-198,583:4-OCH3-ICI-198,583, 4-methoxy-ICI-198,583 Glu-to-Val-ICI-198,583;Valine-ICI-198;583;Glu-to-Sub-ICI-198,583,2-aminosuberic acid-ICI-198,583;7-CH3-ICI-198,583,7-methyl-ICI-198,583;ZD1694,N-[5(N-(3,4-dihydro-2-methyl-4-oxoquinazoline-6-yl-methyl)amino)2-thienyl)]-L-glutamic acid;2-NH2 -ZD1694, 2-amino-ZD1694;BW1843U89, (S)-2[5-(((1,2-dihydro-3-methyl-1-oxobenzo(f)quinazolin-9-yl)methyl)amino-)-1-oxo-2-isoindolinyl]-glutaric acid;LY231514, N-(4-(2-(2-amino-4,7-dihydro-4-oxo-3H-pyrrolo[ 2,3-D]pyrimidine-5-yl)ethyl)-benzoyl]-L-glutamic acid; IAHQ, 5,8-dideazaisofolate; 2-dIAHQ, 2-desamino-IAHQ; 2-CH3-dIAHQ, 2-desamino-2-methyl-IAHQ; 5-d(i)PteGlu, 5-deazaisofolate; N9-CH3-5-d(i)PteGlu, N9-methyl-5-deazaisofolate; N9-CHO-5-d(i)PteGlu, N9-formyl-5-deazaisofolate; AG337, 3,4-dihydro-2-amino-6-methyl-4-oxo-5-(4-pyridylthio)quanazoline; and 2,4-diamino-6[N-(4-(phenysulfonyl)benzyl)ethyl)amino]quinazoline; or their stereoisomers; [5] A composition of item [1] wherein the folate antagonist is selected from methotrexate, larcitrexed, previtrexed, pemetrexed, lometrexol (LMX; 5,10-dideazatetrahydrofolate), cyclopenta[g]quinazoline having a dipeptide ligand, CB3717, CB300945, or stereoisomers thereof such as 6-R, S-BGC945 (ONX-0801), CB300638, and BW1843U89; [6] A composition according to any of items [1] to [5], wherein the gamma polyglutamine oxidized folate antimetabolitic agent contains 4, 5, 2 to 10, 4 to 6, or more than 5 glutamyl groups: [7] A composition according to any of items [1] to [6], wherein the gamma polyglutamine oxoxide antimetabolitic agent is (a) It is a gammatetraglutamine oxoxide antimetabolitic agent, (b) a gamma-pentaglutamine oxoxide antimetabolitic agent, or (c) It is a gamma-hexaglutamine oxidase folate antagonist. composition; [8] A composition according to any of items [1] to [7], wherein the gamma-polyglutamine oxidized folate antimetabolitic agent comprises 1 to 10 glutamyl groups having gamma-carboxyl group bonds; [9] A composition described in any of items [1] to [8], (a) At least two glutamyl groups of the gamma polyglutamine oxoxide antimetabolites are in the L form; (b) Each glutamyl group in the gamma polyglutamine oxoxide antimetabolites is L-type; (c) At least one glutamyl group of the gamma polyglutamine oxoxide antimetabolite is of the D type; (d) The glutamyl groups of each gamma polyglutamate oxidase folate antagonist, other than the glutamyl group of the folate antagonist, are of type D; or (e) At least two glutamyl groups of the gamma polyglutamine oxoxide antimetabolites are L-type and at least one glutamyl group is D-type. Composition:

[10] A composition according to any of items [1] to [9], wherein the polyglutamic acid is linear;

[11] A composition according to any of items [1] to [9], wherein the polyglutamic acid is a branched chain;

[12] Liposome composition containing a gamma polyglutamine oxidase folate antimetabolitic agent as described in any of items [1] to

[11] (Lp-γPANTIFOL);

[13] An Lp-γPANTIFOL composition as described in item

[12] , wherein the polyglutamic acid oxidase antagonist is selected from the following: (a) AG2034, pyritrexime, pralatrexate, GW1843, folate antagonists, and LY309887; or (b) PMX, MTX, RTX, and LTX, and their stereoisomers;

[14] Lp-γPANTIFOL compositions according to item

[12] or

[13] , wherein the polyglutamic acid oxidase antagonist is selected from the following: LV (etoposide), L-leucovorin (L-5-formyltetrahydrofolate); 5-CH3-THF, 5-methyltetrahydrofolate; FA, folic acid; PteGlu, pteroylglutamate (FA); MTX, methotrexate; 2-dMTX, 2-desamino-MTX; 2-CH3-MTX, 2-desamino-2-methyl-MTX; AMT, aminopterin; 2-dAMT, 2-desamino-AMT; 2-CH3-AMT, 2-desamino-2-methyl-AMT; 10-EdAM, 10-ethyl-10-deazaaminopterin; PT523, N Alpha-(4-amino-4-deoxypterol)-N delta-(hemiphthaloyl)-L-ornithine;DDATHF (lometrexol), 5,10-dideaza-5,6,7,8-tetrahydrofolate;5-d(i)H4PteGlu, 5-deaza-5,6,7,8-tetrahydroisofolate;N9-CH3-5-d(i)H4PteGlu, N9-methyl-5-deaza-5,6,7,8-tetrahydroisofolate;5-dPteHCysA,N alpha-(5-deazapteroyl)-L-homocysteinic acid;5-dPteAPBA,N alpha-(5-deazapteroyl)-DL-2-amino-4-phosphonobutyric acid;5-dPteOrn,N Alpha-(5-deazapteroyl)-L-ornithine; 5-dH4PteHCysA,N Alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-L-homocysteic acid; 5-dH4PteAPBA,N Alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-DL-2-amino-4-phosphobutanoic acid; 5-dH4PteOro,N Alpha-(5-dideaza-5,6,7,8-tetrahydropteroyl)-L-ornithine; CB3717,N10-propargyl-5,8-dideazafolate; ICI-198,583,2-desamino-2-methyl-N10-propargyl-5,8-dideazafolate;4-H-ICI-198,583, 4-deoxy-ICI-198,583:4-OCH3-ICI-198,583, 4-methoxy-ICI-198,583 Glu-to-Val-ICI-198,583;Valine-ICI-198;583;Glu-to-Sub-ICI-198,583, 2-aminosuberic acid-ICI-198,583;7-CH3-ICI-198,583,7-methyl-ICI-198,583;ZD1694, N-[5(N-(3,4-dihydro-2-methyl-4-oxoquinazoline-6-yl-methyl (amino)2-thienyl)-L-glutamic acid; 2-NH2-ZD1694, 2-amino-ZD1694; BW1843U89, (S)-2[5-(((1,2-dihydro-3-methyl-1-oxobenzo(f)quinazolin-9-yl)methyl)amino-)-1-oxo-2-isoindlinyl]-glutaric acid; LY231514, N-(4-(2-(2-amino-4,7-dihydroxy Dro-4-oxo-3H-pyrrolo[2,3-D]pyrimidine-5-yl)ethyl)-benzoyl]-L-glutamic acid;IAHQ, 5,8-dideazaisofolate;2-dIAHQ, 2-desamino-IAHQ;2-CH3-dIAHQ, 2-desamino-2-methyl-IAHQ;5-d(i)PteGlu, 5-deazaisofolate;N9-CH3-5-d(i)PteGlu,N9- Methyl-5-deazaisofolate; N9-CHO-5-d(i)PteGlu, N9-formyl-5-deazaisofolate; AG337, 3,4-dihydro-2-amino-6-methyl-4-oxo-5-(4-pyridylthio)quanazoline; and AG377, 2,4-diamino-6[N-(4-(phenysulfonyl)benzyl)ethyl)amino]quinazoline; or their stereoisomers;

[15] Lp-γPANTIFOL compositions according to any of items

[12] to

[14] , wherein the folate antagonist is selected from methotrexate, larcitrexed, previtrexed, pemetrexed, lometrexol (LMX; 5,10-dideazatetrahydrofolate), cyclopenta[g]quinazoline having a dipeptide ligand, CB3717, CB300945, or stereoisomers thereof such as 6-R, S-BGC945 (ONX-0801), CB300638, and BW1843U89;

[16] An Lp-γPANTIFOL composition according to any of items

[12] to

[15] , wherein the liposomes contain a gamma polyglutamic oxidase folate antimetabolitic agent comprising 4, 5, 2 to 10, 4 to 6, or more than 5 gamma glutamyl groups;

[17] An Lp-γPANTIFOL composition according to any of items

[12] to

[16] , wherein the liposome contains a gammatetraglutamine oxidase folate antimetabolitic agent;

[18] An Lp-γPANTIFOL composition according to any of items

[12] to

[16] , wherein the liposome comprises a gammapentaglobulin oxidase folate antimetabolitic agent;

[19] An Lp-γPANTIFOL composition according to any of items

[12] to

[16] , wherein the liposome contains a gamma-hexaglutamine oxidase folate antimetabolitic antagonist;

[20] An Lp-γPANTIFOL composition according to any of items

[12] to

[19] , wherein the gamma-polyglutamine oxidized folate antimetabolitic agent comprises 1 to 10 glutamyl groups having gamma-carboxyl group bonds;

[21] An Lp-γPANTIFOL composition according to any of items

[12] to

[20] , (a) At least two glutamyl groups of the gamma polyglutamine oxoxide antimetabolites are in the L form; (b) Each glutamyl group in the gamma polyglutamine oxoxide antimetabolites is L-type; (c) At least one glutamyl group of the gamma polyglutamine oxoxide antimetabolite is of the D type; (d) The glutamyl groups of each gamma polyglutamate oxidase folate antagonist, other than the glutamyl group of the folate antagonist, are of type D; or (e) At least two glutamyl groups of the gamma polyglutamine oxoxide antimetabolites are L-type and at least one glutamyl group is D-type. Lp-γPANTIFOL composition:

[22] An Lp-γPANTIFOL composition according to any of items

[12] to

[21] , which: (a) At least two glutamyl groups of the gamma polyglutamine oxoxide antimetabolites are in the L form; (b) Each glutamyl group in the gamma polyglutamine oxoxide antimetabolites is L-type; (c) At least one glutamyl group of the gamma polyglutamine oxoxide antimetabolite is of the D type; (d) The glutamyl groups of each gamma polyglutamate oxidase folate antagonist, other than the glutamyl group of the folate antagonist, are of type D; or (e) At least two glutamyl groups of the gamma polyglutamine oxoxide antimetabolites are L-type and at least one glutamyl group is D-type. Lp-γPANTIFOL composition:

[23] An Lp-γPANTIFOL composition according to any of items

[12] to

[22] , wherein the liposomes are pegylated (PLp-γPANTIFOL);

[24] An Lp-γPANTIFOL composition according to any of items

[12] to

[22] , wherein the liposomes are not pegylated;

[25] An Lp-γPANTIFOL composition according to any of items

[12] to

[24] , wherein the liposomes have a diameter in the range of 20 nm to 200 nm;

[26] An Lp-γPANTIFOL composition according to any of items

[12] to

[25] , wherein the liposomes have a diameter in the range of 80 nm to 120 nm;

[27] An Lp-γPANTIFOL composition according to any of items

[12] to

[26] , wherein the liposomes are formed from liposome components;

[28] An Lp-γPANTIFOL composition as described in item

[27] , wherein the liposome component comprises at least one anionic lipid and a neutral lipid;

[29] Lp-γPANTIFOL compositions according to item

[27] or

[28] , wherein the liposome component comprises at least one selected from DSPE; DSPE-PEG; DSPE-PEG-maleimide; HSPC; HSPC-PEG; cholesterol; cholesterol-PEG; and cholesterol-maleimide;

[30] Lp-γPANTIFOL compositions according to any of items

[27] to

[29] , wherein the liposome component comprises at least one selected from the following: Lp-γPANTIFOL compositions; DSPE; DSPE-PEG; DSPE-PEG-FITC; DSPE-PEG-maleimide; cholesterol; and HSPC;

[31] An Lp-γPANTIFOL composition according to any of items

[27] to

[30] , wherein one or more liposome components further comprise a steric stabilizer;

[32] Lp-γPANTIFOL compositions according to item

[31] , wherein the steric stabilizer is polyethylene glycol (PEG); poly-L-lysine (PLL); monosialoganglioside (GM1); poly(vinylpyrrolidone) (PVP); poly(acrylamide) (PAA); poly(2-methyl-2-oxazoline); poly(2-ethyl-2-oxazoline); phosphatidyl polyglycerol; poly[N-(2-hydroxypropyl)methacrylamide]; amphiphilic poly-N-vinylpyrrolidone; L-amino acid-based polymer; oligoglycerin, polyethylene glycol and polypropylene oxide-containing copolymer, poloxamer 188, and polyvinyl alcohol;

[33] An Lp-γPANTIFOL composition as described in item

[32] , wherein the steric stabilizer is PEG, and the PEG has a number-average molecular weight (Mn) of 200 to 5000 daltons;

[34] An Lp-γPANTIFOL composition according to any of items

[12] to

[33] , wherein the liposomes are anionic or neutral;

[35] An Lp-γPANTIFOL composition according to any of items

[12] to

[33] , wherein the liposomes have a zeta potential of zero or less;

[36] An Lp-γPANTIFOL composition according to any of items

[12] to

[33] , wherein the liposomes have a zeta potential of 0 to -150 mV;

[37] An Lp-γPANTIFOL composition according to any of items

[12] to

[33] , wherein the liposomes have a zeta potential of -30 to -50 mV;

[38] An Lp-γPANTIFOL composition according to any of items

[12] to

[33] , wherein the liposomes are cationic;

[39] An Lp-γPANTIFOL composition according to any of items

[12] to

[38] , wherein the liposome has an internal space containing a gamma polyglutamine oxidized folate antimetabolitic antagonist and an aqueous pharmaceutically acceptable carrier;

[40] Lp-γPANTIFOL compositions as described in item

[39] , wherein the pharmaceutically acceptable carrier comprises an isotonic agent such as dextrose, mannitol, glycerol, potassium chloride, or sodium chloride in a concentration greater than 1%;

[41] An Lp-γPANTIFOL composition as described in item

[39] , wherein the aqueous pharmaceutically acceptable carrier is trehalose;

[42] Lp-γPANTIFOL compositions as described in item

[41] , wherein the pharmaceutically acceptable carrier is 1% to 50% trehalose;

[43] An Lp-γPANTIFOL composition according to any of items

[39] to

[42] , wherein the pharmaceutically acceptable carrier comprises a 1% to 50% dextrose solution;

[44] An Lp-γPANTIFOL composition according to any of items

[39] to

[43] , wherein the internal space of the liposome contains 5% dextrose suspended in HEPES buffer solution;

[45] An Lp-γPANTIFOL composition according to any of items

[39] to

[44] , wherein the pharmaceutically acceptable carrier comprises a buffer such as HEPES buffered saline (HBS) or an analogue with a concentration of 1 to 200 mM and a pH of 2 to 8;

[46] An Lp-γPANTIFOL composition according to any of items

[39] to

[45] , wherein the pharmaceutically acceptable carrier comprises sodium acetate and calcium acetate in a total concentration of 50 mM to 500 mM;

[47] An Lp-γPANTIFOL composition according to any of items

[12] to

[46] , wherein the internal space of the liposomes has a pH of 5 to 8 or a pH of 6 to 7, or any range in between;

[48] ​​An Lp-γPANTIFOL composition according to any of items

[12] to

[47] , wherein the liposomes contain less than 500,000 or less than 200,000 gamma polyglutamine oxidized folate antimetabolitic antagonist molecules;

[49] An Lp-γPANTIFOL composition according to any of items

[12] to

[48] , wherein the liposomes contain 10 to 100,000 or any range in between gamma polyglutamine oxidized folate antimetabolitic antagonist molecules;

[50] An Lp-γPANTIFOL composition according to any of items

[12] to

[49] , further comprising a targeting moiety, wherein the targeting moiety has specific affinity for a surface antigen on a target cell of interest;

[51] An Lp-γPANTIFOL composition as described in item

[50] , wherein the targeting portion is bound to one or both of the PEG and outer surface of the liposome, and optionally, the targeting portion is covalently bound to one or both of the PEG and outer surface of the liposome;

[52] An Lp-γPANTIFOL composition according to item

[50] or

[51] , wherein the targeting portion is a polypeptide;

[53] An Lp-γPANTIFOL composition according to any of items

[50] to

[52] , wherein the targeting portion is an antibody or an antigen-binding fragment of an antibody;

[54] An Lp-γPANTIFOL composition according to any of items

[50] to

[53] , wherein the targeting portion is measured by BIACORE® analysis to be 0.5 x 10⁻¹⁰ to 10 x 10⁻¹⁰ -6Lp-γPANTIFOL composition that binds to surface antigens with an equilibrium dissociation constant (Kd) in the range of [value];

[55] An Lp-γPANTIFOL composition according to any of items

[50] to

[54] , wherein the targeting portion specifically binds to 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);

[56] An Lp-γPANTIFOL composition according to any of items

[50] to

[55] , wherein the targeting portion comprises one or more selected from the group consisting of antibodies, humanized antibodies, antigen-binding fragments of antibodies, single-chain antibodies, single-domain antibodies, bispecific antibodies, synthetic antibodies, pegylated antibodies, and multimeric antibodies;

[57] An Lp-γPANTIFOL composition according to any of items

[50] to

[56] , wherein each pegylated liposome contains 1 to 1000 or 30 to 200 targeting moieties;

[58] An Lp-γPANTIFOL composition according to any of items

[39] to

[57] , further comprising one or more of an immunostimulant, a detectable marker, and maleimide, wherein the immunostimulant, the detectable marker, or the maleimide is bound to the PEG or outer surface of a liposome;

[59] An Lp-γPANTIFOL composition according to item

[58] , wherein the immunostimulant is at least one selected from the group consisting of protein immunostimulants, nucleic acid immunostimulants, chemoimmunostimulants, haptens, and adjuvants;

[60] Lp-γPANTIFOL composition as described in item

[58] or

[59] , wherein the immunostimulant is fluorescein, fluorescein isothiocyanate (FITC), DNP, beta-glucan, beta-1,3-glucan, beta-1,6-glucan, resolvin (e.g., D n-6DPA Or D n-3DPAAn Lp-γPANTIFOL composition comprising at least one selected from the group consisting of resolvin D, resolvin E, or T-series resolvins, oxidized low-density lipoproteins (e.g., OXPAC, PGPC), and Toll-like receptor (TLR) modulators such as erythritol lipids (e.g., E5564);

[61] An Lp-γPANTIFOL composition described in any of items

[58] to

[60] , wherein the immunostimulant and the detectable marker are the same;

[62] An Lp-γPANTIFOL composition according to any of items

[58] to

[61] , further comprising a hapten;

[63] Lp-γPANTIFOL composition as described in item

[62] , wherein the hapten comprises one or more of fluorescein or beta-1,6-glucan:

[64] An Lp-γPANTIFOL composition according to any of items

[12] to

[63] , further comprising at least one cryoprotective substance selected from the group consisting of mannitol, trehalose, sorbitol, and sucrose;

[65] Targeted compositions comprising any of the compositions described in items [1] to

[64] ;

[66] Non-targeting compositions comprising any of the compositions described in items [1] to

[49] ;

[67] An Lp-γPANTIFOL composition according to any of items

[12] to

[66] , further comprising carboplatin and / or pembrolizumab;

[68] A pharmaceutical composition comprising a liposomal gamma polyglutamine oxidized folate antimetabolitic agent composition described in any of items

[12] to

[67] ;

[69] A pharmaceutical composition comprising a gamma polyglutamine oxoxide antimetabolitic agent composition described in any of items [1] to [7];

[70] A composition described in any of items [1] to

[69] for use in the treatment of a disease;

[71] Use of any of the compositions described in items [1] to

[70] in the manufacture of a drug for the treatment of a disease;

[72] A method for treating or preventing a disease of which such treatment or prevention is required, comprising the step of administering a composition of any of items [1] to

[70] to the subject;

[73] A method for treating or preventing a disease in a subject requiring such treatment or prevention, comprising administering to a subject the liposomal gamma polyglutamine oxidized folate antimetabolitic composition described in any of

[12] to

[69] ;

[74] A method for killing overgrown cells, comprising the step of bringing the overgrown cells into contact with a composition described in any of items [1] to

[69] ;

[75] A method for killing hyperproliferating cells, comprising the step of contacting the hyperproliferating cells with a liposomal gamma polyglutamine oxidized folate antimetabolitic composition described in any of items

[12] to

[69] ;

[76] Methods relating to item

[74] or

[75] , wherein the overgrowth cells are cancer cells, mammalian cells, and / or human cells;

[77] A method for treating cancer, comprising the step of administering an effective amount of any composition described in item [1] to

[69] to a subject who has or is at risk of having cancer;

[78] A method for treating cancer, comprising the step of administering an effective amount of a liposomal gamma polyglutamine oxidized folate antimetabolitic composition described in any of items

[12] to

[68] to a subject who has or is at risk of having cancer;

[79] Methods according to item

[77] or

[78] , wherein the cancer is a non-hematological malignancy, including, for example, lung cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, head and neck cancer, stomach cancer, gastrointestinal cancer, colorectal cancer, esophageal cancer, cervical cancer, liver cancer, kidney cancer, bile duct cancer, gallbladder cancer, bladder cancer, sarcoma (e.g., osteosarcoma), brain cancer, central nervous system cancer, and melanoma; and a method selected from, for example, hematological malignancies, including leukemia, lymphoma and other B-cell malignancies, myeloma and other plasma cell proliferation disorders;

[80] A method according to item

[77] or

[78] , wherein the cancer is selected from lung cancer, breast cancer, colorectal cancer, pancreatic cancer, gastric cancer, bladder cancer, head and neck cancer, ovarian cancer, and cervical cancer;

[81] A method according to item

[77] or

[78] , wherein the cancer is selected from colorectal cancer, lung cancer, breast cancer, head and neck cancer, and pancreatic cancer;

[82] A method according to item

[77] or

[78] , wherein the cancer is selected from colorectal cancer, breast cancer, ovarian cancer, lung cancer, head and neck cancer, pancreatic cancer, gastric cancer, and mesothelioma;

[83] A method for treating cancer, comprising the step of administering an effective amount of the Lp-γPANTIFOL composition described in any of items

[50] to

[66] to a subject having or at risk of having cancer cells expressing folate receptors bound by the targeting moiety on their surface;

[84] Maintenance therapy for subjects who are or have been receiving cancer therapy, comprising the step of administering an effective amount of any of the compositions described in items [1] to

[69] to a subject who is or has been receiving cancer therapy;

[85] Maintenance therapy for subjects currently receiving or formerly receiving cancer therapy, comprising the step of administering an effective amount of a liposomal gamma polyglutamine oxidative antimetabolitic composition described in any of items

[12] to

[69] to a subject currently receiving or formerly receiving cancer therapy;

[86] A method for treating an immune system disorder comprising the step of administering an effective amount of any composition described in item [1] to

[69] to a subject having or at risk of having an immune system disorder, wherein the immune system disorder is selected from inflammation (e.g., acute and chronic), systemic inflammation, rheumatoid arthritis, inflammatory bowel disease (IBD), Crohn's disease, dermatomyositis / polymyositis, systemic lupus erythematosus, and Takayasu's arteritis, and psoriasis;

[87] A method for treating an immune system disorder comprising the step of administering an effective amount of a liposomal gamma polyglutamine oxidized folate antimetabolitic composition described in any of items [8] to

[69] to a subject having or at risk of having an immune system disorder, wherein the immune system disorder is selected from inflammation (e.g., acute and chronic), systemic inflammation, rheumatoid arthritis, inflammatory bowel disease (IBD), Crohn's disease, dermatomyositis / polymyositis, systemic lupus erythematosus, and Takayasu's arteritis, and psoriasis;

[88] The following treatment methods: (a) A method for treating an infectious disease, comprising the step of administering an effective amount of any composition described in item [1] to

[69] to a subject having or at risk of having an infectious disease; (b) A method for treating an infectious disease, cardiovascular disease, metabolic disease, or another disease, comprising the step of administering an effective amount of any composition described in item [1] to

[69] to a subject having or at risk of having an infectious disease, cardiovascular disease, or another disease, wherein the disease is a member selected from atherosclerosis, cardiovascular disease (CVD), coronary artery disease, myocardial infarction, stroke, metabolic syndrome, gestational trophoblastic disease, and ectopic pregnancy; (c) A method for treating an autoimmune disease, comprising the step of administering an effective amount of any composition described in item [1] to

[69] to a subject having or at risk of having an autoimmune disease; (d) A method for treating rheumatoid arthritis, comprising the step of administering an effective amount of any composition described in item [1] to

[69] to a subject having or at risk of having rheumatoid arthritis; (e) A method for treating an inflammatory condition comprising the step of administering an effective amount of any composition described in item [1] to

[69] to a subject having or at risk of having inflammation, wherein the inflammation is acute, chronic, and / or systemic inflammation; or (f) A method for treating a skin disease, comprising the step of administering an effective amount of any composition described in item [1] to

[69] to a subject who has or is at risk of having a skin disease;

[89] A method for treating an infectious disease, comprising the step of administering an effective amount of a liposomal gamma polyglutamine oxidized folate antimetabolitic composition described in any of items

[12] to

[69] to a subject who has or is at risk of having an infectious disease;

[90] A method for delivering a gamma-polyglutamine oxidase antagonist to a tumor expressing folate receptors on its surface, comprising the step of administering an Lp-γPANTIFOL composition described in any of items [1] to

[69] to a subject having a tumor in an amount that delivers a therapeutically effective dose of the gamma-polyglutamine oxidase antagonist to the tumor;

[91] A method for preparing a gamma polyglutamate oxidase antagonist composition comprising a liposome gamma polyglutamate oxidase antagonist composition described in any of items

[12] to

[69] , comprising the steps of: forming a mixture comprising a liposome component and a gamma polyglutamate oxidase antagonist in solution; homogenizing the mixture in solution to form liposomes; and processing the mixture to form liposomes comprising a gamma polyglutamate oxidase antagonist.

[92] A method for preparing any of the compositions described in items

[12] to

[69] , comprising the steps of: forming a mixture in solution containing a liposome component and a gamma-polyglutamate oxidase antagonist; homogenizing the mixture in solution to form liposomes; processing the mixture to form liposomes that encapsulate and / or contain the gamma-polyglutamate oxidase antagonist; and imparting a targeting moiety on the surface of the liposomes, wherein the targeting moiety has specific affinity for at least one of folate receptor alpha (FR-α), folate receptor beta (FR-β), and folate receptor delta (FR-δ).

[93] Methods according to item

[92] , wherein the processing step comprises one or more steps from thin film hydration, extrusion, in-line mixing, ethanol injection technique, freeze-thaw method, reverse-phase evaporation method, dynamic high-pressure microfluidization, microfluidic mixing, double emulsion, freeze-dried double emulsion, 3D printing, membrane contactor method, and stirring; and / or

[94] A method according to item

[92] , wherein the processing step comprises one or more steps of changing the size of liposomes by one or more steps of extrusion, high-pressure microfluidization, and / or sonication.

[0010] In some embodiments, the Disclosure provides a gamma-polyglutamate folate antimetabolite (γPANTIFOL) composition in which at least two glutamyl residues of the gamma-polyglutamate folate antimetabolite have gamma-carboxyl group bonds. In some embodiments, γPANTIFOL contains 2 to 20, 2 to 15, 2 to 10, 2 to 5, or more than 5 glutamyl groups (including the glutamyl groups of the folate antimetabolite). In some embodiments, the gamma-polyglutamate folate antimetabolite is selected from (a) AG2034, pyritrexime, pralatrexate, GW1843, folate antimetabolite, and LY309887; or (b) PMX, MTX, RTX, and LTX, or their stereoisomers. In some embodiments, the gamma polyglutamate oxidase antagonist is selected from the following: LV (etoposide), L-leucovorin (L-5-formyltetrahydrofolate); 5-CH3-THF, 5-methyltetrahydrofolate; FA, folate; PteGlu, pteroylglutamate (FA); MTX, methotrexate; 2-dMTX, 2-desamino-MTX; 2-CH3-MTX, 2-desamino-2-methyl-MTX; AMT, aminopterin; 2-dAMT, 2-desamino-AMT; 2-CH3-AMT, 2-desamino-2-methyl-AMT; 10-EdAM, 10-ethyl-10-deazaaminopterin; PT523, N alpha-(4-amino-4-deoxypteroyl)-N Delta-(hemiphthaloyl)-L-ornithine;DDATHF (lometrexol), 5,10-dideaza-5,6,7,8-tetrahydrofolate;5-d(i)H4PteGlu, 5-deaza-5,6,7,8-tetrahydroisofolate;N9-CH3-5-d(i)H4PteGlu, N9-methyl-5-deaza-5,6,7,8-tetrahydroisofolate;5-dPteHCysA,N Alpha-(5-deazapteroyl)-L-homocysteic acid;5-dPteAPBA,N Alpha-(5-deazapteroyl)-DL-2-amino-4-phosphonobutyric acid;5-dPteOrn,N Alpha-(5-deazapteroyl)-L-ornithine;5-dH4PteHCysA,N Alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-L-homocysteinic acid;5-dH4PteAPBA,N alpha-(5-dideaza-5,6,7,8-tetrahydropteroyl)-DL-2-amino-4-phosphobutanoic acid; 5-dH4PteOro,N alpha-(5-dideaza-5,6,7,8-tetrahydropteroyl)-L-ornithine; CB3717,N 10-propargyl-5,8-dideaza folate; ICI-198,583, 2-desamino-2-methyl-N 10-propargyl-5,8-dideaza folate; 4-H-ICI-198,583, 4-deoxy-ICI-198,583:4-OCH3-ICI-198,583, 4-methoxy-ICI-198,583 Glu-to-Val-ICI-198,583;Valine-ICI-198;583;Glu-to-Sub-ICI-198,583,2-Aminoseveric acid-ICI-198,583;7-CH3-ICI-198,583,7-Methyl-ICI-198,583;ZD1694,N-[5(N-(3,4-Dihydro-2-methyl-4-oxoquinazoline-6-yl- Methyl)amino)2-thienyl)]-L-glutamic acid; 2-NH2-ZD1694, 2-amino-ZD1694; BW1843U89, (S)-2[5-(((1,2-dihydro-3-methyl-1-oxobenzo(f)quinazolin-9-yl)methyl)amino-)-1-oxo-2-isoindolinyl]-glutaric acid; LY231514, N-(4-(2-(2-amino -4,7-dihydro-4-oxo-3H-pyrrolo[2,3-D]pyrimidine-5-yl)ethyl)-benzoyl]-L-glutamic acid;IAHQ, 5,8-dideazaisofolate;2-dIAHQ, 2-desamino-IAHQ;2-CH3-dIAHQ, 2-desamino-2-methyl-IAHQ;5-d(i)PteGlu, 5-deazaisofolate;N9-CH3-5-d(i) PteGlu, N9-methyl-5-deazaisofolate; N9-CHO-5-d(i)PteGlu, N9-formyl-5-deazaisofolate; AG337, 3,4-dihydro-2-amino-6-methyl-4-oxo-5-(4-pyridylthio)quanazoline; and AG377, 2,4-diamino-6[N-(4-(phenysulfonyl)benzyl)ethyl)amino]quinazoline;or their stereoisomers. In some embodiments, the gamma polyglutamic oxidase antimetabolites are selected from methotrexate, larcitrexed, previtrexed, pemetrexed, lometrexol (LMX; 5,10-dideazatetrahydrofolate), cyclopenta[g]quinazoline with a dipeptide ligand, CB3717, CB300945, or their stereoisomers such as 6-R, S-BGC945 (ONX-0801), CB300638, and BW1843U89. In some embodiments, γPANTIFOL comprises two or more L-type glutamyl groups. In some embodiments, γPANTIFOL comprises a D-type glutamyl group. In further embodiments, γPANTIFOL comprises a D-type glutamyl group and two or more L-type glutamyl groups.

[0011] In one embodiment, the γPANTIFOL composition comprises a chain of three glutamyl groups bonded to a glutamyl group in the folate antagonist (i.e., a γ-tetraglutamate folate antagonist). In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [2] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [3] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [4] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [5] of the Summary of Invention section. In some embodiments, γPANTIFOL is a polyglutamate folate antagonist described in the Summary of Invention section. In some embodiments, the tetraglutamate folate antagonist comprises two or more L-type glutamyl groups. In other embodiments, the tetraglutamine oxidase antagonist contains a D-type glutamyl group. In some embodiments, the tetraglutamine oxidase antagonist contains two or more D-type glutamyl groups. In further embodiments, the tetraglutamine oxidase antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the tetraglutamine oxidase antagonist contains one, two, or three D-type glutamyl groups and three, two, or one L-type glutamyl group, respectively.

[0012] In one embodiment, the γPANTIFOL composition comprises a chain of four γ-glutamyl groups bonded to a glutamyl group in the folate antagonist (i.e., a γ-pentaglutamate folate antagonist). In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [2] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [3] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [4] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [5] of the Summary of Invention section. In some embodiments, γPANTIFOL is a polyglutamate folate antagonist described in the Summary of Invention section. In some embodiments, the pentagultamine oxidase antagonist comprises two or more L-type glutamyl groups. In other embodiments, the pentagultamine oxidase antagonist comprises a D-type glutamyl group. In some embodiments, the pentagultamine oxidase antagonist comprises two or more D-type glutamyl groups. In further embodiments, the pentagultamine oxidase antagonist comprises a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the pentagultamine oxidase antagonist comprises one, two, three, or four D-type glutamyl groups and four, three, two, or one L-type glutamyl group, respectively.

[0013] In one embodiment, the γPANTIFOL composition comprises a chain of five γ-glutamyl groups bonded to a glutamyl group in the folate antagonist (i.e., a γ-hexaglutamate folate antagonist). In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [2] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [3] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [4] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [5] of the Summary of Invention section. In some embodiments, γPANTIFOL is a polyglutamate folate antagonist described in the Summary of Invention section. In some embodiments, the hexaglutamine oxidase antagonist comprises two or more L-type glutamyl groups. In some embodiments, the hexaglutamine oxidase antagonist comprises a D-type glutamyl group. In some embodiments, the hexaglutamine oxidase antagonist comprises two or more D-type glutamyl groups. In further embodiments, the hexaglutamine oxidase antagonist comprises a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the pentaglutamine oxidase antagonist comprises one, two, three, four, or five D-type glutamyl groups and five, four, three, two, or one L-type glutamyl group, respectively.

[0014] In further embodiments, the disclosure provides compositions comprising delivery carriers such as liposomes filled (i.e., encapsulated) and / or otherwise bound thereto with a gamma-polyglutamate folate antagonist, and methods for preparing γPANTIFOL-filled / bound delivery carrier compositions (DV-γPANTIFOL) and methods for using them to deliver gamma-polyglutamate folate antagonists to diseased (e.g., cancerous) cells and / or target cells. These compositions have applications including, but are not limited to, the treatment of diseases including, for example, hyperproliferative diseases such as cancer, immune system disorders such as inflammation and rheumatoid arthritis, and infectious diseases such as HIV, malaria, and schistosomiasis. The γPANTIFOL-filled / bound delivery carrier compositions result in improved efficacy and safety of folate antagonist delivery to cancer cells by providing selective delivery of a higher cytotoxic payload (polyglutamate folate antagonist) compared to the cytotoxicity of folate antagonists (ANTIFOL) administered in monoglutamate form. In some embodiments, the gamma-polyglutamate folate antagonist in DV-γPANTIFOL contains 2 to 20, 2 to 15, 2 to 10, 2 to 5, or more than 5, or more than 20 glutamyl groups (including the glutamyl groups of the folate antagonist). In some embodiments, the delivery carrier contains the polyglutamate folate antagonist described in any of the items [1] to

[11] in the Summary of the Invention section. In some embodiments, the delivery carrier contains the polyglutamate folate antagonist described in the Summary of the Invention section. In some embodiments, the delivery carrier is a liposome described in any of the items

[12] to

[67] in the Summary of the Invention section.

[0015] In further embodiments, the disclosure provides compositions (Lp-γPANTIFOL) comprising liposomes encapsulated (filled) with a gamma-polyglutamate folate antagonist. In some embodiments, the gamma-polyglutamate folate antagonist in Lp-γPANTIFOL comprises 2 to 20, 2 to 15, 2 to 10, 2 to 5, or more than 20 glutamyl groups (including the glutamyl groups of the folate antagonist). In some embodiments, the gamma-polyglutamate folate antagonist encapsulated by the liposome is selected from (a) AG2034, pyritrexime, pralatrexate, GW1843, the folate antagonist, and LY309887; or (b) PMX, MTX, RTX, and LTX, or their stereoisomers. In some embodiments, the liposome-encapsulated gamma-polyglutamic acid folate antimetabolites are selected from the following: LV (etoposide), L-leucovorin (L-5-formyltetrahydrofolate); 5-CH3-THF, 5-methyltetrahydrofolate; FA, folate; PteGlu, pteroylglutamate (FA); MTX, methotrexate; 2-dMTX, 2-desamino-MTX; 2-CH3-MTX, 2-desamino-2-methyl-MTX; AMT, aminopterin; 2-dAMT, 2-desamino-AMT; 2-CH3-AMT, 2-desamino-2-methyl-AMT; 10-EdAM, 10-ethyl-10-deazaaminopterin; PT523, N alpha-(4-amino-4-deoxypteroyl)-N Delta-(hemiphthaloyl)-L-ornithine; DDATHF (lometrexol), 5,10-dideaza-5,6,7,8-tetrahydrofolate; 5-d(i)H4PteGlu, 5-deaza-5,6,7,8-tetrahydroisofolate; N9-CH3-5-d(i)H4PteGlu, N9-methyl-5-deaza-5,6,7,8-tetrahydroisofolate; 5-dPteHCysA, N alpha-(5-deazapteroyl)-L-homocysteine; 5-dPteAPBA, N alpha-(5-deazapteroyl)-DL-2-amino-4-phosphonobutyric acid; 5-dPteOrn, N alpha-(5-deazapteroyl)-L-ornithine;5-dH4PteHCysA,N alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-L-homocysteic acid; 5-dH4PteAPBA,N alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-DL-2-amino-4-phosphobutanoic acid; 5-dH4PteOro,N Alpha-(5-dideaza-5,6,7,8-tetrahydropteroyl)-L-ornithine;CB3717, N10-propargyl-5,8-dideazafolate;ICI-198,583, 2-desamino-2-methyl-N10-propargyl-5,8-dideazafolate;4-H-ICI-198,583, 4-deoxy-ICI-198,583:4-OCH3-ICI-198,583, 4-methoxy-ICI-198,583 Glu-to-Val-ICI-198,583;Valine-ICI-198;583;Glu-to-Sub-ICI-198,583,2-Aminosveric acid-ICI-198,583;7-CH3-ICI-198,583,7-Methyl-ICI-198,583;ZD1694,N-[5(N-(3,4-Dihydro-2-methyl-4- Oxoquinazoline-6-yl-methyl)amino)2-thienyl)]-L-glutamic acid; 2-NH2-ZD1694, 2-amino-ZD1694; BW1843U89, (S)-2[5-(((1,2-dihydro-3-methyl-1-oxobenzo(f)quinazoline-9-yl)methyl)amino-)-1-oxo-2-isoindlinyl]-glutamic acid Taric acid; LY231514, N-(4-(2-(2-amino-4,7-dihydro-4-oxo-3H-pyrrolo[2,3-D]pyrimidine-5-yl)ethyl)-benzoyl]-L-glutamic acid; IAHQ, 5,8-dideazaisofolic acid; 2-dIAHQ, 2-desamino-IAHQ; 2-CH3-dIAHQ, 2-desamino-2-methyl-I AHQ; 5-d(i)PteGlu, 5-deazaisofolate; N9-CH3-5-d(i)PteGlu, N9-methyl-5-deazaisofolate; N9-CHO-5-d(i)PteGlu, N9-formyl-5-deazaisofolate; AG337, 3,4-dihydro-2-amino-6-methyl-4-oxo-5-(4-pyridylthio)quanazoline;and AG377, 2,4-diamino-6[N-(4-(phenisulfonyl)benzyl)ethyl)amino]quinazoline; or stereoisomers thereof. In some embodiments, the gamma polyglutamic oxidase antagonist encapsulated by liposomes is selected from methotrexate, larcitrexed, previtrexed, pemetrexed, lometrexol (LTX; 5,10-dideazatetrahydrofolate), cyclopenta[g]quinazoline having a dipeptide ligand, CB3717, CB300945, or stereoisomers thereof such as 6-R, S-BGC945 (ONX-0801), CB300638, and BW1843U89. In some embodiments, the gamma polyglutamic oxidase antagonist in Lp-γPANTIFOL contains two or more L-type glutamyl groups. In other embodiments, the gamma-polyglutamine oxidase antagonist in Lp-γPANTIFOL contains a D-type glutamyl group. Further embodiments include the gamma-polyglutamine oxidase antagonist in Lp-γPANTIFOL containing a D-type glutamyl group and two or more L-type glutamyl groups.

[0016] In one embodiment, the Lp-γPANTIFOL composition comprises a gamma-polyglutamate folate antagonist containing a chain of three glutamyl groups bonded to a glutamyl group in the folate antagonist (i.e., a tetraglutamate folate antagonist). In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [2] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [3] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [4] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [5] of the Summary of Invention section. In some embodiments, γPANTIFOL is a polyglutamate folate antagonist described in the Summary of Invention section. In some embodiments, the tetraglutamine oxidase antagonist comprises two or more L-type glutamyl groups. In other embodiments, the tetraglutamine oxidase antagonist comprises a D-type glutamyl group. In further embodiments, the tetraglutamine oxidase antagonist comprises a D-type glutamyl group and two or more L-type glutamyl groups.

[0017] In one embodiment, the Lp-γPANTIFOL composition comprises a gamma-polyglutamate folate antagonist containing a chain of four γ-glutamyl groups bonded to a glutamyl group in the folate antagonist (e.g., a γ-pentaglutamate folate antagonist). In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [2] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [3] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [4] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [5] of the Summary of Invention section. In some embodiments, γPANTIFOL is a polyglutamate folate antagonist described in the Summary of Invention section. In some embodiments, the pentaglobulin oxidase antagonist comprises two or more L-type glutamyl groups. In some embodiments, the gamma-pentaglutamine oxidase antagonist comprises a D-type glutamyl group. In further embodiments, the pentaglobulin oxidase antagonist comprises a D-type glutamyl group and two or more L-type glutamyl groups.

[0018] In one embodiment, the Lp-γPANTIFOL composition comprises a gamma-polyglutamate folate antagonist containing a chain of five γ-glutamyl groups bonded to a glutamyl group in the folate antagonist (e.g., a γ-hexaglutamate folate antagonist). In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [2] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [3] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [4] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is a folate antagonist described in item [5] of the Summary of Invention section. In some embodiments, γPANTIFOL is a polyglutamate folate antagonist described in the Summary of Invention section. In some embodiments, the gamma-hexaglutamine oxidase antagonist comprises two or more L-type glutamyl groups. In other embodiments, the gamma-hexaglutamine oxidase antagonist comprises a D-type glutamyl group. In further embodiments, the gamma-hexaglutamine oxidase antagonist comprises a D-type glutamyl group and two or more L-type glutamyl groups.

[0019] In some embodiments, the Lp-γPANTIFOL composition is cationic. In some embodiments, the Lp-γPANTIFOL liposomes are cationic and have a diameter in the range of 20 nm to 200 nm, 30 nm to 175 nm, or 50 nm to 150 nm, or any range in between. In further embodiments, the Lp-γPANTIFOL liposomes are cationic and have a diameter in the range of 30 nm to 175 nm or 50 nm to 150 nm, or any range in between. In further embodiments, the Lp-γPANTIFOL liposomes have a diameter in the range of 80 nm to 120 nm, or any range in between. In some embodiments, the cationic Lp-γPANTIFOL composition comprises at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of liposome-encapsulated gamma-tetraglutamine oxidase folate antagonist. In some embodiments, the cationic Lp-γPANTIFOL comprises at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of liposome-encapsulated gamma-pentaglutamine oxidase folate antagonist. In other embodiments, Lp-γPANTIFOL comprises at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of liposome-encapsulated gamma-hexaglutamine-oxidized folate antagonist. In further embodiments, the liposome-encapsulated gamma-polyglutamine-oxidized folate antagonist is present in HEPES buffer within the liposomes.

[0020] In other embodiments, the Lp-γPANTIFOL composition is anionic or neutral. In some embodiments, the Lp-γPANTIFOL composition is cationic. In some embodiments, the Lp-γPANTIFOL liposomes are anionic or neutral and have a diameter in the range of 20 nm to 200 nm, 30 nm to 175 nm, or 50 nm to 150 nm, or any range in between. In further embodiments, the Lp-γPANTIFOL liposomes are anionic or neutral and have a diameter in the range of 30 nm to 175 nm or 50 nm to 150 nm, or any range in between. In further embodiments, the Lp-γPANTIFOL liposomes are anionic or neutral and the composition has a diameter in the range of 80 nm to 120 nm, or any range in between. In some embodiments, the Lp-γPANTIFOL liposomes are anionic and have a diameter in the range of 20 nm to 200 nm, 30 nm to 175 nm, or 50 nm to 150 nm, or any range in between. In further embodiments, the Lp-γPANTIFOL liposomes are anionic and have a diameter in the range of 30 nm to 175 nm or 50 nm to 150 nm, or any range in between. In further embodiments, the Lp-γPANTIFOL liposomes are anionic and the composition has a diameter in the range of 80 nm to 120 nm, or any range in between. In some embodiments, the Lp-γPANTIFOL liposomes are neutral and have a diameter in the range of 20 nm to 200 nm, 30 nm to 175 nm, or 50 nm to 150 nm, or any range in between. In further embodiments, the Lp-γPANTIFOL liposomes are neutral and have a diameter in the range of 30 nm to 175 nm or 50 nm to 150 nm, or any range in between. In further embodiments, the Lp-γPANTIFOL liposomes are neutral and the composition has a diameter in the range of 80 nm to 120 nm, or any range in between.In some embodiments, the anionic or neutral Lp-γPANTIFOL composition comprises at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of liposome-encapsulated gamma-tetraglutamine oxidase folate antagonist. In other embodiments, the Lp-γPANTIFOL composition comprises at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of liposome-encapsulated gamma-hexaglutamine-oxidized folate antagonist. In further embodiments, the liposome-encapsulated gamma-polyglutamine-oxidized folate antagonist is present in HEPES buffer within the liposomes.

[0021] In further embodiments, the liposomal gamma-polyglutamine oxidized folate antimetabolitic agent composition is pegylated (PLp-γPANTIFOL).

[0022] In some embodiments, the liposomal gamma-polyglutamine oxidase antagonist composition is untargeted (NTLp-γPANTIFOL). That is, the NTLp-γPANTIFOL composition does not have specific affinity for an epitope expressed on the surface of the target cell of interest (e.g., an epitope on a surface antigen). In some embodiments, the NTLp-γPANTIFOL composition does not contain a targeting moiety. In further embodiments, the untargeted liposomal gamma-polyglutamine oxidase antagonist composition is pegylated (NTPLp-γPANTIFOL).

[0023] In other embodiments, the liposomal gamma polyglutamine oxidized folate antimetabolite composition is targeted (TLp-γPANTIFOL). That is, the TLp-γPANTIFOL composition includes a targeting moiety having specific affinity for an epitope (surface antigen) on the target cell of interest. In some embodiments, TLp-γPANTIFOL or TPLp-γPANTIFOL is not covalently bound to the liposome. In other embodiments, the targeting moiety of TLp-γPANTIFOL or TPLp-γPANTIFOL is bound to one or both of the PEG and outer surface of the liposome. In some embodiments, the targeting moiety of TLp-γPANTIFOL or TPLp-γPANTIFOL is covalently bound to the liposome. The functions of the targeting moiety of TLp-γPANTIFOL and / or TPLp-γPANTIFOL compositions include, but are not limited to, targeting liposomes to target cells of interest in vivo or in vitro; interacting with surface antigens having specific affinity for the targeting moiety; and delivering the liposome payload (γPANTIFOL) to the cells. Preferred targeting moieties are known in the art and are not limited to, antibodies, antigen-binding antibody fragments, scaffold proteins, polypeptides, and peptides. In some embodiments, the targeting moiety is a polypeptide. In further embodiments, the targeting moiety is a polypeptide comprising at least 3, 5, 10, 15, 20, 30, 40, 50, or 100 amino acid residues.

[0024] Targeted liposomal gamma-polyglutamine-oxidized folate antimetabolites (TLp-γPANTIFOL and TPLp-γPANTIFOL) offer further improvements to the efficacy and safety profiles of folate antimetabolites by specifically delivering gamma-polyglutamine-oxidized (e.g., γ-pentaglutamine-oxidized and / or γ-hexaglutamine-oxidized) folate antimetabolites to target cells such as cancer cells. In further embodiments, the targeted liposomal gamma-polyglutamine-oxidized folate antimetabolites are pegylated (TPLp-γPANTIFOL). In some embodiments, the targeting moieties of TLp-γPANTIFOL and TPLp-γPANTIFOL are bound to one or both of the PEG and outer surfaces of the liposome. In some embodiments, the targeting moieties of TLp-γPANTIFOL and TPLp-γPANTIFOL are bound to the liposome via covalent bonds (γPANTIFOL). The functions of the targeting moiety of TLp-γPANTIFOL and / or TPLp-γPANTIFOL compositions include, but are not limited to, targeting liposomes to target cells of interest in vivo or in vitro; interacting with surface antigens having specific affinity for the targeting moiety; and delivering the liposome payload (γPANTIFOL) to the cells. Preferred targeting moieties are known in the art and are not limited to, antibodies, antigen-binding antibody fragments, scaffold proteins, polypeptides, and peptides. In some embodiments, the targeting moiety is a polypeptide. In further embodiments, the targeting moiety is a polypeptide comprising at least 3, 5, 10, 15, 20, 30, 40, 50, or 100 amino acid residues.

[0025] In some embodiments, the targeting portion of TLp-γPANTIFOL or TPLp-γPANTIFOL is an antibody or an antigen-binding antibody fragment. In further embodiments, the targeting portion includes one or more of the following: an antibody, a humanized antibody, an antigen-binding fragment of an antibody, a single-chain antibody, a single-domain antibody, a bispecific antibody, a synthetic antibody, a pegylated antibody, and a multimeric antibody. In some embodiments, the targeting portion of TLp-γPANTIFOL or TPLp-γPANTIFOL has specific affinity for epitopes selectively expressed on target cells, such as tumor cells, compared to normal or non-tumor cells. In some embodiments, the targeting portion has specific affinity for epitopes on tumor cell surface antigens that are present on tumor cells but not present or difficult to access on non-tumor cells. In some embodiments, the targeting portion is measured by BIACORE® analysis to be 0.5 x 10⁻⁶ -10 ~10x10 -6 It binds to the target epitope with an equilibrium dissociation constant (Kd) within the specified range.

[0026] In certain embodiments, the TLp-γPANTIFOL or TPLp-γPANTIFOL targeting moiety comprises a polypeptide that specifically binds to a folate receptor. In some embodiments, the targeting moiety is an antibody or an antigen-binding antibody fragment. In some embodiments, the folate receptor bound by the targeting moiety specifically binds to one or more folate receptors selected from folate receptor alpha (FR-α, FOLR1), folate receptor beta (FR-β, FOLR2), and folate receptor delta (FR-δ, FOLR4). In some embodiments, the folate receptor bound by the targeting moiety is folate receptor alpha (FR-α). In some embodiments, the folate receptor bound by the targeting moiety is folate receptor beta (FR-β). In some embodiments, the targeting moiety specifically binds to both FR-α and FR-β.

[0027] In further embodiments, the Lp-γPANTIFOL composition comprises one or more immunostimulants, detectable markers, and maleimides, disposed on at least one of the PEG or outer surfaces of the liposome. In some embodiments, the liposome γPANTIFOL composition (e.g., Lp-γPANTIFOL, PLp-γPANTIFOL, NTLp-γPANTIFOL, NTPLp-γPANTIFOL, TLp-γPANTIFOL, or TPLp-γPANTIFOL) is cationic. In other embodiments, the liposome γPANTIFOL composition (e.g., Lp-γPANTIFOL, PLp-γPANTIFOL, NTLp-γPANTIFOL, NTPLp-γPANTIFOL, TLp-γPANTIFOL, or TPLp-γPANTIFOL) is anionic or neutral. In further embodiments, the liposomes of the liposome-γPANTIFOL composition (e.g., Lp-γPANTIFOL, PLp-γPANTIFOL, NTLp-γPANTIFOL, NTPLp-γPANTIFOL, TLp-γPANTIFOL, or TPLp-γPANTIFOL) have a diameter in the range of 20 nm to 200 nm, or any range in between. In further embodiments, the liposomes of the liposome-γPANTIFOL composition have a diameter in the range of 80 nm to 120 nm, or any range in between. In some embodiments, the liposome-γPANTIFOL composition is pegylated (e.g., PLp-γPANTIFOL, NTPLp-γPANTIFOL, or TPLp-γPANTIFOL). In some embodiments, the liposome-γPANTIFOL composition includes a targeting moiety (e.g., TLp-γPANTIFOL or TPLp-γPANTIFOL). In further embodiments, the liposomal γPANTIFOL composition is pegylated and targeted (e.g., TPLp-γPANTIFOL). In some embodiments, the liposomal γPANTIFOL composition comprises a gamma-polyglutamate oxidase antagonist containing 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomal γPANTIFOL composition comprises a gamma-tetraglutamate oxidase antagonist.In some embodiments, the liposomal γPANTIFOL composition comprises a gamma-pentaglutamine oxidase antagonist. In some embodiments, the liposomal γPANTIFOL composition comprises a gamma-hexaglutamine oxidase antagonist. In some embodiments, the liposomal composition comprises a gamma-polyglutamine oxidase antagonist as described in any of sections [1] to

[11] of the Summary of the Invention. In some embodiments, the liposome comprises the liposomal composition described in any of items

[11] to

[69] of the Summary of the Invention. In some embodiments, the composition comprises a gamma-polyglutamine oxidase antagonist as described in the Summary of the Invention.

[0028] In further embodiments, the liposomal γPANTIFOL composition (i.e., Lp-γPANTIFOL such as PLp-γPANTIFOL, NTLp-γPANTIFOL, NTPLp-γPANTIFOL, TLp-γPANTIFOL, or TPLp-γPANTIFOL) contains at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of the liposomal encapsulated gamma polyglutamate oxidase antagonist. In some embodiments, the liposomal γPANTIFOL composition contains 1% to 98.5% of the liposomal encapsulated gamma polyglutamate oxidase antagonist. In further embodiments, the liposomal γPANTIFOL composition comprises at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of a liposomal encapsulated gamma polyglutamate oxidase antagonist containing 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomal γPANTIFOL composition comprises 1% to 98.5% of a liposomal encapsulated gamma polyglutamate oxidase antagonist containing 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomal γPANTIFOL composition contains at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of liposomal encapsulated gamma-tetraglutamine oxidase folate antagonist. In some embodiments, the liposomal γPANTIFOL composition contains 1% to 98.5% of liposomal encapsulated gamma-polyglutamine oxidase folate antagonist. In some embodiments, the liposomal γPANTIFOL composition contains at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of liposomal encapsulated gamma-pentaglutamine oxidase folate antagonist. In some embodiments, the liposomal γPANTIFOL composition contains 1% to 98.5% liposomal encapsulated gammapentaglobulin oxidase folate antagonist.In some embodiments, the liposomal γPANTIFOL composition comprises at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of the liposomal encapsulated gamma-hexaglutamine oxidase antagonist. In some embodiments, the liposomal γPANTIFOL composition comprises 1% to 98.5% of the liposomal encapsulated gamma-pentaglutamine oxidase antagonist. In some embodiments, the liposomal composition comprises the gamma-polyglutamine oxidase antagonist as described in any of sections [1] to

[11] of the Outline of the Invention. In some embodiments, the liposome comprises the liposomal composition described in any of sections

[11] to

[69] of the Outline of the Invention. In some embodiments, the composition comprises the gamma-polyglutamine oxidase antagonist as described in the Outline of the Invention or in the figures herein.

[0029] Liposome compositions comprising γPANTIFOL-encapsulated liposomes are also provided. In some embodiments, the liposome composition comprises a pegylated γPANTIFOL composition. In some embodiments, the liposome composition comprises a γPANTIFOL composition linked to or otherwise conjugated to a targeting moiety. In further embodiments, the liposome composition comprises a pegylated γPANTIFOL composition linked to or otherwise conjugated to a targeting moiety. In some embodiments, the liposome composition comprises γPANTIFOL comprising 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposome composition comprises a gamma-tetraglutamine-oxidized folate antagonist. In some embodiments, the liposome composition comprises a gamma-pentaglutamine-oxidized folate antagonist. In other embodiments, the liposome composition comprises a gamma-hexaglutamine-oxidized folate antagonist. In some embodiments, the polyglutamine-oxidized folate antagonist is a folate antagonist described in item [2] of the Summary of the Invention section. In some embodiments, the polyglutamate folate antagonist is the folate antagonist described in item [3] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is the folate antagonist described in item [4] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is the folate antagonist described in item [5] of the Summary of Invention section. In some embodiments, γPANTIFOL is the polyglutamate folate antagonist described in the Summary of Invention section.

[0030] In some embodiments, the liposome composition comprises liposome γPANTIFOL (e.g., Lp-γPANTIFOL, PLp-γPANTIFOL, NTLp-γPANTIFOL, NTPLp-γPANTIFOL, TLp-γPANTIFOL, and TPLp-γPANTIFOL). In some embodiments, the liposome γPANTIFOL is pegylated (e.g., NTPLp-γPANTIFOL and TPLp-γPANTIFOL). In some embodiments, the pharmaceutical composition comprises γPANTIFOL containing 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the pharmaceutical composition comprises a gammatetraglutamine oxidase antagonist. In some embodiments, the pharmaceutical composition comprises a gammapentaglutamine oxidase antagonist. In other embodiments, the pharmaceutical composition comprises a gammahexaglutamine oxidase antagonist. In some embodiments, the polyglutamate-oxidized folate antagonist is the folate antagonist described in item [2] of the Summary of Invention section. In some embodiments, the polyglutamate-oxidized folate antagonist is the folate antagonist described in item [3] of the Summary of Invention section. In some embodiments, the polyglutamate-oxidized folate antagonist is the folate antagonist described in item [4] of the Summary of Invention section. In some embodiments, the polyglutamate-oxidized folate antagonist is the folate antagonist described in item [5] of the Summary of Invention section. In some embodiments, γPANTIFOL is the polyglutamate-oxidized folate antagonist described in the Summary of Invention section. In some embodiments, liposomal γPANTIFOL includes a targeting moiety having specific affinity for an antigen epitope on the surface of a target cell of interest, such as cancer cells (e.g., TLp-γPANTIFOL or TPLp-γPANTIFOL). In further embodiments, the liposome composition comprises pegylated liposome-γPANTIFOL and further comprises a targeting moiety having specific affinity for an antigen epitope on the surface of a target cell of interest, such as cancer cells (e.g., TPLp-γPANTIFOL). In some embodiments, the liposome composition comprises a cationic liposome-γPANTIFOL.In other embodiments, the liposome composition comprises liposome γPANTIFOL which is anionic or neutral. In further embodiments, the liposome composition comprises liposome γPANTIFOL having a diameter in the range of 20 nm to 200 nm, or any range in between. In further embodiments, the liposome γPANTIFOL has a diameter in the range of 80 nm to 120 nm, or any range in between.

[0031] Pharmaceutical compositions are also provided that include a gamma-polyglutamine oxidase inhibitor (γPANTIFOL) containing a delivery carrier such as liposomal γPANTIFOL. In some embodiments, the pharmaceutical composition includes a pegylated γPANTIFOL composition. In some embodiments, the pharmaceutical composition includes a γPANTIFOL composition linked to or otherwise conjugated to a targeting moiety. In further embodiments, the pharmaceutical composition includes a pegylated γPANTIFOL composition linked to or otherwise conjugated to a targeting moiety. In some embodiments, the pharmaceutical composition includes γPANTIFOL containing 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the pharmaceutical composition includes a gamma-tetraglutamine oxidase inhibitor. In some embodiments, the pharmaceutical composition includes a gamma-pentaglutamine oxidase inhibitor. In other embodiments, the pharmaceutical composition includes a gamma-hexaglutamine oxidase inhibitor. In some embodiments, the gamma-polyglutamate oxidase antagonist is a polyglutamate oxidase antagonist described in any of the items [1] to

[11] in the summary section of the invention.

[0032] In some embodiments, the pharmaceutical composition comprises liposome-γPANTIFOL (e.g., Lp-γPANTIFOL, PLp-γPANTIFOL, NTLp-γPANTIFOL, NTPLp-γPANTIFOL, TLp-γPANTIFOL, and TPLp-γPANTIFOL). In some embodiments, the liposome-γPANTIFOL composition is pegylated (e.g., NTPLp-γPANTIFOL and TPLp-γPANTIFOL). In some embodiments, the liposome-γPANTIFOL comprises a targeting moiety having specific affinity for an antigen epitope on the surface of a target cell of interest, such as a cancer cell (e.g., TLp-γPANTIFOL or TPLp-γPANTIFOL). In further embodiments, the pharmaceutical composition comprises pegylated liposome-γPANTIFOL, further comprising a targeting moiety having specific affinity for an antigen epitope on the surface of a target cell of interest, such as a cancer cell (e.g., TPLp-γPANTIFOL). In some embodiments, the pharmaceutical composition comprises a cationic liposomal γPANTIFOL. In other embodiments, the pharmaceutical composition comprises an anionic or neutral liposomal γPANTIFOL. In further embodiments, the pharmaceutical composition comprises a liposomal γPANTIFOL having a diameter in the range of 20 nm to 200 nm, or any range in between. In further embodiments, the liposomal γPANTIFOL composition has a diameter in the range of 80 nm to 120 nm, or any range in between. In some embodiments, the pharmaceutical composition comprises a γPANTIFOL comprising 4, 5, 2 to 10, 4 to 6, or more than 5 glutamyl groups. In some embodiments, the pharmaceutical composition comprises a gamma-tetraglutamine oxidase folate antagonist. In some embodiments, the pharmaceutical composition comprises a gamma-pentaglutamine oxidase folate antagonist. In other embodiments, the pharmaceutical composition comprises a gamma-hexaglutamine oxidase folate antagonist. In some embodiments, the composition comprises a gamma-polyglutamine oxidase folate antagonist described in any of items [1] to

[11] of the Summary of the Invention section. In some embodiments, the pharmaceutical composition includes a liposome composition described in any of the items

[11] to

[69] in the summary section of the invention.In some embodiments, the composition comprises a gamma-polyglutamine oxidase folate antagonist described in the summary section of the invention.

[0033] In further embodiments, the disclosure provides a method for modulating cell activation, chemokine production, or metabolic activity, the method comprising the step of contacting cells with a composition comprising a gamma-polyglutamine oxidase antagonist (γPANTIFOL) composition. In some embodiments, the cells to be contacted are mammalian cells. In further embodiments, the cells to be contacted are human cells. In some embodiments, the cells to be contacted are hyperproliferating cells. In further embodiments, the cells are immune cells. In some embodiments, the method is carried out in vivo. In other embodiments, the method is carried out in vitro. In some embodiments, γPANTIFOL comprises 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the γPANTIFOL composition comprises a gamma-tetraglutamine oxidase antagonist. In some embodiments, the γPANTIFOL composition comprises a gamma-pentaglutamine oxidase antagonist. In other embodiments, the γPANTIFOL composition comprises a gamma-hexaglutamine oxidase antagonist. In some embodiments, the composition comprises a gamma-polyglutamine oxidase antagonist described in any of items [1] to

[11] of the Summary of the Invention section. In some embodiments, the pharmaceutical composition comprises a liposomal composition described in any of items

[11] to

[69] of the Summary of the Invention section. In some embodiments, the composition comprises a gamma-polyglutamine oxidase antagonist described in the Summary of the Invention section or in the figures herein.

[0034] In further embodiments, the disclosure provides a method for modulating cell activation, chemokine production, or metabolic activity, the method comprising the step of contacting cells with liposomes containing a gamma-polyglutamine oxidase inhibitor (γPANTIFOL) composition. In some embodiments, the cells to be contacted are mammalian cells. In further embodiments, the cells to be contacted are human cells. In some embodiments, the cells to be contacted are hyperproliferating cells. In further embodiments, the cells are immune cells. In some embodiments, the method is carried out in vivo. In other embodiments, the method is carried out in vitro. In some embodiments, γPANTIFOL contains 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the γPANTIFOL composition contains a gamma-tetraglutamine oxidase inhibitor. In some embodiments, the γPANTIFOL composition contains a gamma-pentaglutamine oxidase inhibitor. In other embodiments, the γPANTIFOL composition contains a gamma-hexaglutamine oxidase inhibitor. In some embodiments, the polyglutamate folate antagonist includes the folate antagonist described in item [2] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is the folate antagonist described in item [3] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is the folate antagonist described in item [4] of the Summary of Invention section. In some embodiments, the polyglutamate folate antagonist is the folate antagonist described in item [5] of the Summary of Invention section. In some embodiments, γPANTIFOL is the polyglutamate folate antagonist described in the Summary of Invention section.

[0035] In further embodiments, the present disclosure provides a method for killing cells, the method comprising the step of contacting cells with a composition comprising a gamma-polyglutamate oxidized folate antimetabolite (γPANTIFOL) composition. In some embodiments, the cells to be contacted are mammalian cells. In further embodiments, the cells to be contacted are human cells. In some embodiments, the cells to be contacted are overgrowth cells. In further embodiments, the overgrowth cells are cancer cells. In further embodiments, the cancer cells to be contacted are primary cells, or cells derived from cell lines obtained from / derived from cancers selected from, for example, lung cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, head and neck cancer, gastric cancer, gastrointestinal cancer, colorectal cancer, esophageal cancer, cervical cancer, liver cancer, kidney cancer, bile duct cancer, gallbladder cancer, bladder cancer, sarcoma (e.g., osteosarcoma), brain cancer, central nervous system cancer, and melanoma; and, for example, leukemia, lymphoma and other B-cell malignancies, myeloma and other plasmacytic dysplasia or cachexia. In some embodiments, the cancer is selected from breast cancer, advanced head and neck cancer, lung cancer, gastric cancer, osteosarcoma, non-Hodgkin lymphoma (NHL), acute lymphoblastic leukemia (ALL), mycosis fungoides (cutaneous T-cell lymphoma), choriocarcinoma, chorioadenoma, non-leukemic meningeal carcinomatosis, soft tissue sarcoma (tendonoid, invasive fibromatosis), bladder cancer, and central nervous system (CNS) cancer. In some embodiments, the cancer cells to be contacted are primary cells or cells derived from cell lines obtained from / derived from lung cancer (e.g., NSCLC or mesothelioma). In some embodiments, the cancer cells to be contacted are primary cells or cells derived from cell lines obtained from / derived from breast cancer (e.g., HER2++ or triple-negative breast cancer). In some embodiments, the cancer cells to be contacted are primary cells or cells derived from cell lines obtained from / derived from colorectal cancer. In some embodiments, the cancer cells to be contacted are primary cells or cells derived from cell lines obtained from / derived from ovarian cancer. In some embodiments, the cancer cells to be contacted are primary cells obtained from / derived from endometrial cancer or cells derived from a cell line. In some embodiments, the cancer cells to be contacted are primary cells obtained from / derived from pancreatic cancer or cells derived from a cell line.In some embodiments, the cancer cells to be contacted are primary cells obtained from / derived from liver cancer or cells derived from a cell line. In some embodiments, the cancer cells to be contacted are primary cells obtained from / derived from head and neck cancer or cells derived from a cell line. In some embodiments, the cancer cells to be contacted are primary cells obtained from / derived from osteosarcoma or cells derived from a cell line. In some embodiments, the method is carried out in vivo. In other embodiments, the method is carried out in vitro. In some embodiments, γPANTIFOL contains 4, 5, 2-10, 4-6, or more than 5 γ-glutamyl groups. In some embodiments, the γPANTIFOL composition contains a gamma-tetraglutamine oxidase antagonist. In some embodiments, the γPANTIFOL composition contains a gamma-pentaglutamine oxidase antagonist. In other embodiments, the γPANTIFOL composition contains a gamma-hexaglutamine oxidase antagonist. In some embodiments, the gamma-polyglutamate oxidase antagonist is a polyglutamate oxidase antagonist described in any of the items [1] to

[11] in the Summary of the Invention section.

[0036] In further embodiments, the present disclosure provides a method for killing cells, the method comprising the step of contacting cells with liposomes containing a gamma polyglutamate oxidase antagonist (Lp-γPANTIFOL, such as PLp-γPANTIFOL, NTLp-γPANTIFOL, NTPLp-γPANTIFOL, TLp-γPANTIFOL, or TPLp-γPANTIFOL). In some embodiments, the cells to be contacted are mammalian cells. In further embodiments, the cells to be contacted are human cells. In some embodiments, the cells to be contacted are overgrowth cells. In further embodiments, the overgrowth cells to be contacted are cancer cells. In further embodiments, cancer cells are primary cells or cells derived from cell lines obtained from / derived from cancers selected from, for example, lung cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, head and neck cancer, gastric cancer, gastrointestinal cancer, colorectal cancer, esophageal cancer, cervical cancer, liver cancer, kidney cancer, bile duct cancer, gallbladder cancer, bladder cancer, sarcoma (e.g., osteosarcoma), brain cancer, central nervous system cancer, and melanoma; and, for example, leukemia, lymphoma and other B-cell malignancies, myeloma and other plasmacytic dysplasia or cachexia. In some embodiments, the cells are primary cells or cells derived from cell lines obtained from / derived from cancers selected from breast cancer, advanced head and neck cancer, lung cancer, gastric cancer, osteosarcoma, non-Hodgkin lymphoma (NHL), acute lymphoblastic leukemia (ALL), mycosis fungoides (cutaneous T-cell lymphoma), choriocarcinoma, chorioadenoma, non-leukemic meningeal carcinomatosis, soft tissue sarcoma (tendonoid, invasive fibromatosis), bladder cancer, and central nervous system (CNS) cancers. In some embodiments, the contacted cancer cells are primary cells or cells derived from cell lines obtained from / derived from lung cancer (e.g., NSCLC or mesothelioma). In some embodiments, the contacted cancer cells are primary cells or cells derived from cell lines obtained from / derived from breast cancer (e.g., HER2++ or triple-negative breast cancer). In some embodiments, the contacted cancer cells are primary cells or cells derived from cell lines obtained from / derived from colorectal cancer. In some embodiments, the cancer cells to be contacted are primary cells obtained from / derived from ovarian cancer or cells derived from a cell line.In some embodiments, the cancer cells to be contacted are primary cells or cells derived from a cell line obtained from / originating endometrial cancer. In some embodiments, the cancer cells to be contacted are primary cells or cells derived from a cell line obtained from / originating pancreatic cancer. In some embodiments, the cancer cells to be contacted are primary cells or cells derived from a cell line obtained from / originating liver cancer. In some embodiments, the cancer cells to be contacted are primary cells or cells derived from a cell line obtained from / originating head and neck cancer. In some embodiments, the cancer cells to be contacted are primary cells or cells derived from a cell line obtained from / originating osteosarcoma. In some embodiments, the method is carried out in vivo. In other embodiments, the method is carried out in vitro. In some embodiments, the liposomes contain γPANTIFOL containing 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomes contain a gammatetraglutamine oxidase antagonist. In some embodiments, the liposomes contain a gammapentaglutamine oxidase antagonist. In other embodiments, the liposomes comprise a gamma-hexaglutamine oxidase antagonist. In some embodiments, the gamma-polyglutamine oxidase antagonist is a polyglutamine oxidase antagonist described in any of the items [1] to

[11] in the Summary of Invention section. In some embodiments, γPANTIFOL is a polyglutamine oxidase antagonist described in the Summary of Invention section. In some embodiments, the liposomes comprise a liposome described in any of the items

[12] to

[67] in the Summary of Invention section.

[0037] In further embodiments, the present disclosure provides a method for treating cancer, the method comprising the step of administering an effective amount of a delivery carrier (e.g., an antibody-immune complex or liposome) containing a gamma-polyglutamine oxidized folate antimetabolite to a subject having or at risk of having cancer. In some embodiments, the delivery carrier is an antibody-containing immune complex (e.g., including a full-length IgG antibody, a bispecific antibody, or scFv). In some embodiments, the delivery carrier is a liposome (e.g., Lp-γPANTIFOL such as PLp-γPANTIFOL, NTLp-γPANTIFOL, NTPLp-γPANTIFOL, TLp-γPANTIFOL, or TPLp-γPANTIFOL). In some embodiments, the delivery carrier to be administered is pegylated. In some embodiments, the delivery carrier to be administered is not pegylated. In further embodiments, the delivery carrier to be administered includes a targeting moiety having specific affinity for an antigen epitope on the surface of cancer cells.In further embodiments, the delivery carrier includes a targeting moiety that specifically binds to a cell surface antigen selected from the following: GONMB, TACSTD2 (TROP2), CEACAM5, EPCAM, folate receptor (e.g., folate receptor-α, folate receptor-β, or folate receptor-δ), mucin 1 (MUC-1), MUC-6, STEAP1, mesothelin, nectin 4, ENPP3, guanylyl cyclase C (GCC), SLC44A4, NaPi2b, CD70 (TNFSF7), CA9 (carbonic anhydrase), 5T4 (TPBG), SLTRK6, SC-16, tissue factor, LIV-1 (ZIP6), CGEN-15027, p-cadherin, fibronectin Extradomain B (ED-B), VEGFR2 (CD309), tenascin, collagen IV, periostin, endothelin receptor, HER2, HER3, EGFR, IGFR-1, EGFRvIII, CD2, CD3, CD4, CD5, CD6, CD11, CD11a, CD15, CD18, CD19, CD20, CD22, CD26, CD27L, CD30, CD33, CD34, CD37, CD38, CD40, CD44, CD56, CD70, CD74, CD79, CD79b, CD105, CD133, CD138, cripto, CD38, EphA receptor, EphB receptor, EphA2, integrin (for example, integrin α). v β3, α v β5, or α vβ6), C242 antigen, Apo2, PSGR, NGEP, PSCA, TMEFF2, Endoglin, PSMA, CD98, CD56, CanAg, and CALLA. In some embodiments, the delivery carrier includes a targeting moiety that specifically binds to a cell surface antigen that is determined to originate from or be expressed on a specific target cancer (tumor), such as a neoantigen. In some embodiments, the targeting moiety specifically binds to a cell surface antigen that is determined to originate from or be expressed on a specific target tumor, such as a neoantigen. In some embodiments, the targeting moiety is an antibody or an antigen-binding antibody fragment. In some embodiments, the delivered delivery carrier includes γPANTIFOL containing 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the delivered delivery carrier includes a gamma-tetraglutamine oxidase antagonist. In some embodiments, the delivered delivery carrier includes a gamma-pentaglutamine oxidase antagonist. In some embodiments, the delivered delivery carrier includes a L-gamma-polyglutamine oxidase antagonist. In some embodiments, the administered delivery carrier comprises two, three, four, five, or more than five L-gamma-glutamyl groups. In some embodiments, the administered delivery carrier comprises a D-gamma-gamma-polyglutamate oxidase antagonist. In some embodiments, the administered delivery carrier comprises two, three, four, five, or more than five D-gamma-glutamyl groups. In some embodiments, the administered delivery carrier comprises L and D-gamma-gamma-polyglutamate oxidase antagonists. In some embodiments, the administered delivery carrier comprises two, three, four, five, or more than five L-gamma-glutamyl groups and two, three, four, five, or more than five D-gamma-glutamyl groups. In some embodiments, cancer is selected from non-hematological malignancies, such as lung cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, head and neck cancer, stomach cancer, gastrointestinal cancer, colorectal cancer, esophageal cancer, cervical cancer, liver cancer, kidney cancer, bile duct cancer, gallbladder cancer, bladder cancer, sarcoma, brain cancer, central nervous system cancer, and melanoma; and hematological malignancies, such as leukemia, lymphoma and other B-cell malignancies, myeloma and other plasmacytotic dysplasia or cachexia.In some embodiments, the cancer is selected from breast cancer, advanced head and neck cancer, lung cancer, gastric cancer, osteosarcoma, non-Hodgkin lymphoma (NHL), acute lymphoblastic leukemia (ALL), mycosis fungoides (cutaneous T-cell lymphoma), choriocarcinoma, chorioadenoma, non-leukemic meningeal carcinomatosis, soft tissue sarcoma (tendonoid, invasive fibromatosis), bladder cancer, and central nervous system (CNS) cancer. In some embodiments, the cancer is lung cancer (e.g., NSCLC or mesothelioma). In some embodiments, the cancer is breast cancer (e.g., HER2++ or triple-negative breast cancer). In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is endometrial cancer. In some embodiments, the cancer is pancreatic cancer. In some embodiments, the cancer is liver cancer. In some embodiments, the cancer is head and neck cancer. In some embodiments, the cancer is osteosarcoma. In some embodiments, the delivery carrier to be administered comprises γPANTIFOL containing 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the delivery carrier to be administered comprises a gammatetraglutamine oxidase antagonist. In some embodiments, the delivery carrier to be administered comprises a gammapentaglutamine oxidase antagonist. In other embodiments, the delivery carrier to be administered comprises a gammahexaglutamine oxidase antagonist. In some embodiments, the delivery carrier to be administered comprises a polyglutamine oxidase antagonist described in any of the items [1] to

[11] in the Summary of the Invention section. In some embodiments, the delivery carrier comprises a polyglutamine oxidase antagonist described in the Summary of the Invention section. In some embodiments, the delivery carrier to be administered is a liposome composition comprising a polyglutamine oxidase antagonist described in any of the items [1] to

[11] in the Summary of the Invention section. In some embodiments, γPANTIFOL is a polyglutamic oxidase antagonist described in the Summary of the Invention section. In some embodiments, the liposome composition comprises a liposome described in any of the items

[12] to

[67] in the Summary of the Invention section.

[0038] In a further embodiment, the present disclosure provides a method of treating cancer, the method comprising administering to a subject having or at risk of having cancer an effective amount of liposomes comprising a gamma-polyglutamylated folate antagonist (e.g., Lp-γPANTIFOL such as PLp-γPANTIFOL, NTLp-γPANTIFOL, NTPLp-γPANTIFOL, TLp-γPANTIFOL or TPLp-γPANTIFOL). In some embodiments, the liposomes are pegylated. In some embodiments, the liposomes are not pegylated. In a further embodiment, the liposomes comprise a targeting moiety having a specific affinity for an epitope of an antigen on the surface of cancer cells. In a further embodiment, the liposomes comprise a targeting moiety that specifically binds to a cell surface antigen selected from the following: GONMB, TACSTD2 (TROP2), CEACAM5, EPCAM, folate receptor (e.g., folate receptor-α, folate receptor-β or folate receptor-δ), mucin 1 (MUC-;1), MUC-6, STEAP1, mesothelin, nectin 4, ENPP3, guanylyl cyclase C (GCC), SLC44A4, NaPi2b, CD70 (TNFSF7), CA9 (carbonic anhydrase), 5T4 (TPBG), SLTRK6, SC-16, tissue factor, LIV-1 (ZIP6), CGEN-15027, P-cadherin, fibronectin extra domain B (ED-B), VEGFR2 (CD309), tenascin, collagen IV, periostin, endothelin receptor, HER2, HER3, EGFR, IGFR-1, EGFRvIII, CD2, CD3, CD4, CD5, CD6, CD11, CD11a, CD15, CD18, CD19, CD2(), CD22, CD26, CD27L, CD30, CD();3, CD34, CD37, CD38, CD40, CD44, CD56, CD70, CD74, CD79, CD79b, CD105, CD133, CD138, cripto, CD38, EphA receptor, EphB receptor, EphA2, integrin (e.g., integrin α v β3, α v β5, or α vβ6), C242 antigen, Apo2, PSGR, NGEP, PSCA, TMEFF2, Endoglin, PSMA, CD98, CD56, CanAg, and CALLA. This also includes the use of cancer stem cell targeting moieties such as those targeting CD34, CD133 and CD44, CD138, and CD15. In some embodiments, the targeting moiety is an antibody or antigen-binding antibody fragment. In some embodiments, the liposome contains γPANTIFOL containing 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposome contains a gamma-tetraglutamine oxidase antagonist. In some embodiments, the liposome contains a gamma-pentaglutamine oxidase antagonist. In other embodiments, the liposome contains a gamma-hexaglutamine oxidase antagonist. In some embodiments, the polyglutamate oxidase folate antagonist comprises a folate antagonist described in any of the items [1] to

[11] in the Summary of the Invention section. In some embodiments, γPANTIFOL is a polyglutamate oxidase folate antagonist described in the Summary of the Invention section. In some embodiments, the liposome composition comprises a liposome described in any of the items

[12] to

[67] in the Summary of the Invention section. In some embodiments, the liposome comprises an L-gamma polyglutamate oxidase folate antagonist. In some embodiments, the liposome comprises two, three, four, five, or more than five L-gamma-glutamyl groups. In some embodiments, the liposome comprises a D-gamma polyglutamate oxidase folate antagonist. In some embodiments, the liposome comprises two, three, four, five, or more than five D-gamma-glutamyl groups. In some embodiments, the administered liposome comprises two, three, four, five, or more than five L-gamma-glutamyl groups. In some embodiments, the liposomes contain L and D gamma-gamma-glutamyl polyglutamate oxidase antagonists. In some embodiments, the liposomes contain 2, 3, 4, 5, or 5, or more than 5, L-gamma-glutamyl groups and 2, 3, 4, 5, or more than 5, D-gamma-glutamyl groups.In some embodiments, cancer is selected from lung cancer (e.g., non-small cell lung cancer), pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, head and neck cancer, stomach cancer, gastrointestinal cancer, colorectal cancer, esophageal cancer, cervical cancer, liver cancer, kidney cancer, bile duct cancer, gallbladder cancer, bladder cancer, sarcoma (e.g., osteosarcoma), brain cancer, central nervous system cancer, melanoma, and hematological malignancies (e.g., leukemia or lymphoma). In some embodiments, cancer is selected from breast cancer, advanced head and neck cancer, lung cancer, stomach cancer, osteosarcoma, non-Hodgkin lymphoma (NHL), acute lymphoblastic leukemia (ALL), mycosis fungoides (cutaneous T-cell lymphoma), choriocarcinoma, chorioadenoma, non-leukemic leukemic meningocarcinoma, soft tissue sarcoma (tendonoid, invasive fibromatosis), bladder cancer, and central nervous system (CNS) cancer. In some embodiments, the cancer is lung cancer (e.g., NSCLC or mesothelioma). In some embodiments, the cancer is breast cancer (e.g., HER2++ or triple-negative breast cancer). In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer cells are ovarian cancer. In some embodiments, the cancer is endometrial cancer. In some embodiments, the cancer is pancreatic cancer. In some embodiments, the cancer is liver cancer. In some embodiments, the cancer is head and neck cancer. In some embodiments, the cancer is osteosarcoma.

[0039] In further embodiments, the present disclosure provides a method for treating cancer, the method comprising administering an effective amount of a liposome composition comprising a gamma polyglutamine oxoxide antimetabolitic agent and liposomes having a targeting moiety having a specific affinity for an epitope of an antigen on the surface of a cancer tumor, to a subject having or at risk of having cancer. In some embodiments, the liposomes include a targeting moiety that specifically binds to a cell surface antigen selected from the following: GONMB, TACSTD2 (TROP2), CEACAM5, EPCAM, folate receptor (e.g., folate receptor-α, folate receptor-β, or folate receptor-δ), mucin 1 (MUC-1), MUC-6, STEAP1, mesothelin, nectin 4, ENPP3, guanylyl cyclase C (GCC), SLC44A4, NaPi2b, CD70 (TNFSF7), CA9 (carbonic anhydrase), 5T4 (TPBG), SLTRK6, SC-16, tissue factor, LIV-1 (ZIP6), CGEN-15027, p-cadherin, fibronectar Tin extradomain B (ED-B), VEGFR2 (CD309), tenascin, collagen IV, periostin, endothelin receptor, HER2, HER3, EGFR, IGFR-1, EGFRvIII, CD2, CD3, CD4, CD5, CD6, CD11, CD11a, CD15, CD18, CD19, CD20, CD22, CD26, CD27L, CD30, CD33, CD34, CD37, CD38, CD40, CD44, CD56, CD70, CD74, CD79, CD79b, CD105, CD133, CD138, cripto, CD38, EphA receptor, EphB receptor, EphA2, integrin (e.g., integrin α) v β3, α v β5, or α vβ6), C242 antigen, Apo2, PSGR, NGEP, PSCA, TMEFF2, Endoglin, PSMA, CD98, CD56, CanAg, and CALLA. In some embodiments, the liposome includes a targeting moiety that specifically binds to a cell surface antigen(s) of a particular target tumor, such as a neoantigen, or is judged to be derived from or expressed on such a tumor. In some embodiments, the targeting moiety includes an antibody or antigen-binding antibody fragment. In some embodiments, the liposome includes γPANTIFOL containing 4, 5, 2-10, 4-6, or more than 5 γ-glutamyl groups. In some embodiments, the liposome includes a gamma-tetraglutamine oxidase inhibitor. In some embodiments, the liposome includes a gamma-pentaglutamine oxidase inhibitor. In other embodiments, the polyglutamine oxidase inhibitor includes a gamma-hexaglutamine oxidase inhibitor. In some embodiments, the polyglutamic acid folate antagonist is a folate antagonist described in any of the items [1] to

[11] in the Summary of the Invention section. In some embodiments, γPANTIFOL is a polyglutamic acid folate antagonist described in the Summary of the Invention section. In some embodiments, the liposome composition comprises a liposome described in any of the items

[12] to

[67] in the Summary of the Invention section. In some embodiments, the liposome comprises γPANTIFOL comprising an L-type γ-glutamyl group. In some embodiments, the liposome comprises γPANTIFOL comprising a D-type γ-glutamyl group. In some embodiments, the liposome comprises γPANTIFOL comprising at least one L-type γ-glutamyl group and at least one D-type γ-glutamyl group. In some embodiments, the cancer is selected from lung cancer (e.g., non-small cell lung cancer), pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, head and neck cancer, stomach cancer, gastrointestinal cancer, colorectal cancer, esophageal cancer, cervical cancer, liver cancer, kidney cancer, bile duct cancer, gallbladder cancer, bladder cancer, sarcoma (e.g., osteosarcoma), brain cancer, central nervous system cancer, melanoma, and hematological malignancies (e.g., leukemia or lymphoma).In some embodiments, the cancer is selected from breast cancer, advanced head and neck cancer, lung cancer, gastric cancer, osteosarcoma, non-Hodgkin lymphoma (NHL), acute lymphoblastic leukemia (ALL), mycosis fungoides (cutaneous T-cell lymphoma), choriocarcinoma, chorioadenoma, non-leukemic meningeal carcinomatosis, soft tissue sarcoma (tendonoid, invasive fibromatosis), bladder cancer, and central nervous system (CNS) cancer. In some embodiments, the cancer is lung cancer (e.g., NSCLC or mesothelioma). In some embodiments, the cancer is breast cancer (e.g., HER2++ or triple-negative breast cancer). In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is endometrial cancer. In some embodiments, the cancer is pancreatic cancer. In some embodiments, the cancer is liver cancer. In some embodiments, the cancer is head and neck cancer. In some embodiments, the cancer is osteosarcoma.

[0040] In some embodiments, the administered liposome composition comprises pegylated liposomes (e.g., TPLp-γPANTIFOL). In some embodiments, the administered liposome composition comprises non-pegylated liposomes. In some embodiments, the liposomes of the administered liposome composition comprise γPANTIFOL containing 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomes of the administered liposome composition comprise a gammatetraglutamine oxidase antagonist. In some embodiments, the liposomes of the administered liposome composition comprise a gammapentaglutamine oxidase antagonist. In other embodiments, the liposomes of the administered liposome composition comprise a gammahexaglutamine oxidase antagonist. In some embodiments, the liposomes comprise a polyglutamine oxidase antagonist as described in any of items [1]-

[11] of the Summary of the Invention section. In some embodiments, γPANTIFOL is a polyglutamic oxidase antagonist described in the Summary of the Invention section. In some embodiments, the liposome composition comprises a liposome described in any of items

[12] to

[67] in the Summary of the Invention section. In some embodiments, the liposome composition is administered to treat cancers selected from lung cancer (e.g., non-small cell lung cancer), pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, head and neck cancer, gastric cancer, gastrointestinal cancer, colorectal cancer, esophageal cancer, cervical cancer, liver cancer, kidney cancer, bile duct cancer, gallbladder cancer, bladder cancer, sarcoma (e.g., osteosarcoma), brain cancer, central nervous system cancer, melanoma, myeloma and other plasma cell dysplasia or cachexia, as well as leukemia, lymphoma and other B-cell malignancies. In some embodiments, the liposome composition is administered to treat cancers selected from breast cancer, advanced head and neck cancer, lung cancer, gastric cancer, osteosarcoma, non-Hodgkin lymphoma (NHL), acute lymphoblastic leukemia (ALL), mycosis fungoides (cutaneous T-cell lymphoma), choriocarcinoma, chorioadenoma, non-leukemic meningeal carcinomatosis, soft tissue sarcomas (tendonoid sarcoma, invasive fibromatosis), bladder cancer, and central nervous system (CNS) cancers. In some embodiments, the liposome composition is administered to treat lung cancer (e.g., NSCLC or mesothelioma).In some embodiments, the liposomal composition is administered to treat breast cancer (e.g., HER2++ or triple-negative breast cancer). In some embodiments, the liposomal composition is administered to treat colorectal cancer. In some embodiments, the liposomal composition is administered to treat ovarian cancer. In some embodiments, the liposomal composition is administered to treat endometrial cancer. In some embodiments, the liposomal composition is administered to treat pancreatic cancer. In some embodiments, the liposomal composition is administered to treat liver cancer. In some embodiments, the liposomal composition is administered to treat head and neck cancer. In some embodiments, the liposomal composition is administered to treat osteosarcoma.

[0041] In further embodiments, the Disclosure provides a method for treating cancer, comprising the step of administering an effective amount of a liposome composition to a subject having or at risk of having cancer expressing folate receptors on its cell surface, wherein the liposome composition comprises liposomes comprising (a) a gamma-polyglutamate oxidase antagonist (γPANTIFOL) and (b) a targeting moiety having a specific binding affinity to folate receptors. In some embodiments, the targeting moiety has a specific binding affinity to folate receptor alpha (FR-α), folate receptor beta (FR-β), and / or folate receptor delta (FR-δ). In some embodiments, the targeting moiety has a specific binding affinity to folate receptor alpha (FR-α) and folate receptor beta (FR-β). In some embodiments, the administered liposome composition comprises pegylated liposomes (e.g., TPLp-γPANTIFOL). In some embodiments, the administered liposome composition comprises non-pegylated liposomes. In some embodiments, the liposomes of the administered liposome composition contain γPANTIFOL containing 4, 5, 2-10, 4-6, or more than 5 γ-glutamyl groups. In some embodiments, the liposomes of the administered liposome composition contain a gamma-tetraglutamine oxidase antagonist. In some embodiments, the liposomes of the administered liposome composition contain a gamma-pentaglutamine oxidase antagonist. In other embodiments, the liposomes of the administered liposome composition contain a gamma-hexaglutamine oxidase antagonist. In some embodiments, the liposomes contain a polyglutamine oxidase antagonist as described in any of the items [1]-

[11] in the Summary of the Invention section. In some embodiments, γPANTIFOL is a polyglutamine oxidase antagonist as described in the Summary of the Invention section. In some embodiments, the liposome composition contains liposomes as described in any of the items

[12] -

[67] in the Summary of the Invention section.In some embodiments, the liposome composition is administered to treat non-hematological malignancies, including, for example, lung cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, head and neck cancer, gastric cancer, gastrointestinal cancer, colorectal cancer, esophageal cancer, cervical cancer, liver cancer, kidney cancer, bile duct cancer, gallbladder cancer, bladder cancer, sarcomas (e.g., osteosarcoma), brain cancer, central nervous system cancer, and melanoma; and cancers selected from hematological malignancies, such as leukemia, lymphoma and other B-cell malignancies, myeloma and other plasmacytotic dysplasia or cachexia. In some embodiments, the liposome composition is administered to treat cancers selected from breast cancer, advanced head and neck cancer, lung cancer, gastric cancer, osteosarcoma, non-Hodgkin lymphoma (NHL), acute lymphoblastic leukemia (ALL), mycosis fungoides (cutaneous T-cell lymphoma), choriocarcinoma, chorioadenoma, non-leukemic meningeal carcinomatosis, soft tissue sarcomas (tendonoid sarcoma, invasive fibromatosis), bladder cancer, and central nervous system (CNS) cancers. In some embodiments, the liposome composition is administered to treat lung cancer (e.g., NSCLC or mesothelioma). In some embodiments, the liposome composition is administered to treat breast cancer (e.g., HER2++ or triple-negative breast cancer). In some embodiments, the liposome composition is administered to treat colorectal cancer. In some embodiments, the liposome composition is administered to treat ovarian cancer. In some embodiments, the liposome composition is administered to treat endometrial cancer. In some embodiments, the liposome composition is administered to treat pancreatic cancer. In some embodiments, the liposomal composition is administered to treat liver cancer. In some embodiments, the liposomal composition is administered to treat head and neck cancer. In some embodiments, the liposomal composition is administered to treat osteosarcoma.

[0042] In further embodiments, the present disclosure provides a method for maintenance therapy of cancer, the method comprising the step of administering an effective amount of a liposomal composition (Lp-γPANTIFOL) containing a gamma-polyglutamate oxidase antagonist to a subject who is or has been receiving cancer therapy. In some embodiments, the liposomal composition to be administered is PLp-γPANTIFOL, NTLp-γPANTIFOL, NTPLp-γPANTIFOL, TLp-γPANTIFOL, or TPLp-γPANTIFOL. In some embodiments, the liposomes of the liposomal composition to be administered comprise pegylated liposomes (e.g., PLp-γPANTIFOL, NTPLp-γPANTIFOL, or TPLp-γPANTIFOL). In some embodiments, the liposomal composition to be administered comprises targeted liposomes (e.g., TLp-γPANTIFOL or TPLp-γPANTIFOL). In some embodiments, the administered liposome composition comprises pegylated liposomes (e.g., TPLp-γPANTIFOL) containing a targeting moiety. In some embodiments, the liposomes of the administered liposome composition comprise a gamma-polyglutamate oxidase antagonist comprising 4, 5, 2-10, 4-6, or more than 5 γ-glutamyl groups. In some embodiments, the liposomes of the administered liposome composition comprise a gamma-tetraglutamate oxidase antagonist. In some embodiments, the liposomes of the administered liposome composition comprise a gamma-pentaglutamate oxidase antagonist. In some embodiments, the liposomes of the administered liposome composition comprise a gamma-hexaglutamate oxidase antagonist. In some embodiments, the liposome composition comprises liposomes comprising a gamma-polyglutamate as described in any of items [1]-

[11] of the Summary of the Invention section. In some embodiments, γPANTIFOL is a polyglutamic oxidase antagonist described in the Summary of the Invention section. In some embodiments, the liposome composition comprises a liposome described in any of the items

[12] to

[67] in the Summary of the Invention section.

[0043] In further embodiments, the disclosure provides a method for treating an immune system disorder, the method comprising the step of administering an effective amount of a liposomal composition comprising liposomes containing a gamma-polyglutamine oxidized folate antimetabolitic antagonist (e.g., Lp-γPANTIFOL, PLp-γPANTIFOL, NTLp-γPANTIFOL, NTPLp-γPANTIFOL, TLp-γPANTIFOL, or TPLp-γPANTIFOL) to a subject having or at risk of having an immune system disorder. In some embodiments, the liposomal composition is administered to treat an autoimmune disease. In further embodiments, the liposomal composition is administered to treat rheumatoid arthritis. In another embodiment, the liposomal composition is administered to treat inflammation. In some embodiments, the immune system disorder is selected from inflammation (e.g., acute and chronic), systemic inflammation, rheumatoid arthritis, inflammatory bowel disease (IBD), Crohn's disease, dermatomyositis / polymyositis, systemic lupus erythematosus, Takayasu's arteriovenous disease, and psoriasis. In some embodiments, the administered liposome composition comprises pegylated liposomes (e.g., PLp-γPANTIFOL, NTPLp-γPANTIFOL, or TPLp-γPANTIFOL). In some embodiments, the administered liposome composition comprises targeted liposomes (e.g., TLp-γPANTIFOL or TPLp-γPANTIFOL) that have a targeting moiety having specific affinity for a surface antigen on a target cell of interest (e.g., an immune cell). In further embodiments, the administered liposome composition comprises pegylated liposomes containing a targeting moiety (e.g., TPLp-γPANTIFOL). In some embodiments, the liposomes of the administered liposome composition comprise a gamma-polyglutamic acid oxidase antagonist comprising 4, 5, 2-10, 4-6, or more than 5 γ-glutamyl groups. In some embodiments, the liposomes of the administered liposome composition comprise a gamma-tetraglutamic acid oxidase antagonist. In some embodiments, the liposomes of the administered liposome composition contain a gamma-pentaglutamine oxidase antagonist. In other embodiments, the liposomes of the administered liposome composition contain a gamma-hexaglutamine oxidase antagonist.In some embodiments, the liposome composition comprises liposomes containing a gamma polyglutamate as described in any of the items [1] to

[11] in the Summary of the Invention section. In some embodiments, γPANTIFOL is a polyglutamic oxidase folate antagonist as described in the Summary of the Invention section. In some embodiments, the liposome composition comprises liposomes as described in any of the items

[12] to

[67] in the Summary of the Invention section.

[0044] In further embodiments, the Disclosure provides a method for treating an autoimmune disease, the method comprising the step of administering an effective amount of a liposomal composition comprising liposomes containing a gamma-polyglutamic oxidase antimetabolitic agent (e.g., Lp-γPANTIFOL, PLp-γPANTIFOL, NTLp-γPANTIFOL, NTPLp-γPANTIFOL, TLp-γPANTIFOL, or TPLp-γPANTIFOL) to a subject having or at risk of having an autoimmune disease. In some embodiments, the autoimmune disease is rheumatoid arthritis. In some embodiments, the autoimmune disease is a disease or disorder selected from inflammatory bowel disease (IBD), Crohn's disease, systemic lupus erythematosus, and psoriasis. In some embodiments, the autoimmune disease is a disease or disorder selected from the following: Addison's disease, alopecia areata, ankylosing spondylitis, autoimmune hepatitis, autoimmune mumps, diabetes mellitus (type 1), dystrophic epidermolysis bullosa, epididymitis, glomerulonephritis, Graves' disease, Guillain-Barré syndrome, Hashimoto's disease, hemolytic anemia, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, pemphigus vulgaris, psoriasis, rheumatic fever, rheumatoid arthritis, sarcoidosis, scleroderma, Sjögren's syndrome, spondyloarthritis, thyroiditis, vasculitis, vitiligo, myxedema, pernicious anemia, ulcerative colitis. In some embodiments, the liposome composition to be administered comprises pegylated liposomes (e.g., PLp-γPANTIFOL, NTPLp-γPANTIFOL, or TPLp-γPANTIFOL). In some embodiments, the administered liposome composition comprises targeted liposomes (e.g., TLp-γPANTIFOL or TPLp-γPANTIFOL) having a targeting moiety that has specific affinity for a surface antigen on a target cell of interest (e.g., an immune cell). In further embodiments, the administered liposome composition comprises pegylated liposomes (e.g., TPLp-γPANTIFOL) containing a targeting moiety. In some embodiments, the liposomes of the administered liposome composition comprise a gamma-polyglutamine oxidase antagonist containing 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomes of the administered liposome composition comprise a gamma-tetraglutamine oxidase antagonist.In some embodiments, the liposomes of the administered liposome composition contain a gamma-pentaglutamine oxidase antagonist. In other embodiments, the liposomes of the administered liposome composition contain a gamma-hexaglutamine oxidase antagonist. In some embodiments, the liposomes contain a polyglutamine oxidase antagonist as described in any of the items [1] to

[11] in the Summary of the Invention section. In some embodiments, γPANTIFOL is a polyglutamine oxidase antagonist as described in the Summary of the Invention section. In some embodiments, the liposome composition contains a liposome as described in any of the items

[12] to

[67] in the Summary of the Invention section.

[0045] In further embodiments, the present disclosure provides a method for treating an inflammatory disease, the method comprising the step of administering an effective amount of a liposomal composition comprising liposomes containing a gamma-polyglutamate oxidase antagonist (e.g., Lp-γPANTIFOL, PLp-γPANTIFOL, NTLp-γPANTIFOL, NTPLp-γPANTIFOL, TLp-γPANTIFOL, or TPLp-γPANTIFOL) to a subject having or at risk of having an inflammatory disease. In some embodiments, the inflammatory disease is a disease selected from acute inflammation, chronic inflammation, systemic inflammation, rheumatoid arthritis, inflammatory bowel disease (IBD), Crohn's disease, dermatomyositis / polymyositis, and systemic lupus erythematosus. In some embodiments, the inflammatory disease is a disease selected from rheumatoid arthritis or other arthritis (e.g., acute arthritis, acute gouty arthritis, bacterial arthritis, chronic inflammatory arthritis, osteoarthritis, infectious arthritis, juvenile arthritis, fungal arthritis, neuropathic arthritis, polyarthritis, proliferative arthritis, psoriatic arthritis, venereal arthritis, viral arthritis), connective tissue inflammation, pelvic inflammatory disease, acne, psoriasis, actinomycosis, dysentery, biliary cirrhosis, Lyme disease, heat rash, Stevens-Johnson syndrome, mumps, pemphigus vulgaris, and blastomycosis. In some embodiments, the inflammatory disease is inflammatory bowel disease. Inflammatory bowel disease is a chronic inflammatory disease of the gastrointestinal tract, including, but not limited to, Crohn's disease, ulcerative colitis, and unclassified colitis. In some embodiments, the administered liposome composition comprises pegylated liposomes (e.g., PLp-γPANTIFOL, NTPLp-γPANTIFOL, or TPLp-γPANTIFOL). In some embodiments, the administered liposome composition comprises targeted liposomes (e.g., TLp-γPANTIFOL or TPLp-γPANTIFOL) that have a targeting moiety having specific affinity for a surface antigen on a target cell of interest (e.g., an immune cell). In further embodiments, the administered liposome composition comprises pegylated liposomes containing a targeting moiety (e.g., TPLp-γPANTIFOL).In some embodiments, the liposomes of the administered liposome composition contain a gammapentavalent glutamate oxidase antagonist containing 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomes of the administered liposome composition contain a gammatetraglutamine oxidase antagonist. In some embodiments, the liposomes of the administered liposome composition contain a gammapentavalent glutamate oxidase antagonist. In other embodiments, the liposomes of the administered liposome composition contain a gammahexaglutamine oxidase antagonist. In some embodiments, the liposomes contain a polyglutamine oxidase antagonist as described in any of the items [1]-

[11] in the Summary of the Invention section. In some embodiments, γPANTIFOL is a polyglutamine oxidase antagonist as described in the Summary of the Invention section. In some embodiments, the liposome composition contains a liposome as described in any of the items

[12] -

[67] in the Summary of the Invention section.

[0046] This disclosure also provides a method for delivering a gamma-polyglutamate oxidase inhibitor to a target inflammatory site, the method comprising administering to an inflammatory subject a composition comprising a gamma-polyglutamate oxidase inhibitor (L-γPANTIFOL) and a targeting moiety having specific binding affinity to an epitope on a cell surface antigen located at the site of inflammation or otherwise influencing inflammation (e.g., through pro-inflammatory cytokine production). In some embodiments, the targeting moiety to be administered is bound to a delivery carrier. In some embodiments, the delivery carrier is an antibody or an antigen-binding fragment of an antibody. In further embodiments, the delivery carrier is a liposome. In further embodiments, the antibody, antigen-binding antibody fragment, or liposome is a pegylated liposome (e.g., TPLp-γPANTIFOL). In some embodiments, the composition to be administered comprises a gamma-polyglutamate oxidase inhibitor comprising 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the administered composition comprises a gammatetraglutamine oxidase antagonist. In some embodiments, the administered composition comprises a gammapentaglutamine oxidase antagonist. In other embodiments, the administered composition comprises a gammahexaglutamine oxidase antagonist. In some embodiments, γPANTIFOL is a polyglutamine oxidase antagonist described in any of the items [1] to

[11] in the Summary of the Invention section. In some embodiments, γPANTIFOL is a polyglutamine oxidase antagonist described in the Summary of the Invention section. In some embodiments, the delivery carrier is a liposome described in any of the items

[12] to

[67] in the Summary of the Invention section.

[0047] This disclosure also provides a method for delivering a gamma-polyglutamate oxidase inhibitor to a tumor and / or cancer cell, the method comprising administering to a subject having a tumor a composition comprising a gamma-polyglutamate oxidase inhibitor (L-γPANTIFOL) and a targeting moiety having a specific binding affinity to an epitope on a surface antigen of tumor cells or cancer cells. In some embodiments, the targeting moiety to be administered is bound to a delivery carrier. In some embodiments, the delivery carrier is an antibody or an antigen-binding fragment of an antibody. In further embodiments, the delivery carrier is a liposome. In further embodiments, the antibody, antigen-binding antibody fragment, or liposome is a pegylated liposome (e.g., TPLp-γPANTIFOL). In some embodiments, the composition to be administered comprises a gamma-polyglutamate oxidase inhibitor comprising 4, 5, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the composition to be administered comprises a gamma-tetraglutamate oxidase inhibitor. In some embodiments, the administered composition comprises a gamma-pentaglutamine oxidase folate antagonist. In other embodiments, the administered composition comprises a gamma-hexaglutamine oxidase folate antagonist. In some embodiments, the administered composition is a folate antagonist described in any of items [1] to

[11] in the Summary of the Invention section. In some embodiments, γPANTIFOL is a polyglutamine oxidase folate antagonist described in the Summary of the Invention section. In some embodiments, the administered composition comprises liposomes described in any of items

[12] to

[67] in the Summary of the Invention section.

[0048] In further embodiments, the Disclosure provides a method for preparing a liposome composition comprising a liposome gamma-polyglutamine oxidase antagonist (γPANTIFOL) composition, the method comprising the steps of: forming a mixture comprising liposome components and the γ-polyglutamine oxidase antagonist in solution; homogenizing the mixture in solution to form liposomes; and processing the mixture to form liposomes comprising the polyglutamine oxidase antagonist. In some embodiments, the gamma-polyglutamine oxidase antagonist comprises 4, 5, 2-10, 4-6, or more than 5 γ-glutamyl groups. In some embodiments, γPANTIFOL comprises a gamma-tetraglutamine oxidase antagonist. In some embodiments, γPANTIFOL comprises a gamma-pentaglutamine oxidase antagonist. In other embodiments, γPANTIFOL comprises a gamma-hexaglutamine oxidase antagonist. In some embodiments, γPANTIFOL is a polyglutamic acid folate antagonist described in any of the items [1] to

[11] in the Summary of the Invention section. In some embodiments, γPANTIFOL is a polyglutamic acid folate antagonist described in the Summary of the Invention section. In some embodiments, the liposome composition comprises liposomes described in any of the items

[12] to

[67] in the Summary of the Invention section.

[0049] In one embodiment, the present disclosure provides a kit comprising a folate antimetabolite gamma polyglutamate composition and / or a γPANTIFOL delivery carrier such as a liposome containing γPANTIFOL and the γPANTIFOL immune complex described herein (e.g., ADC). [Brief explanation of the drawing]

[0050] [Figure 1A-1N]The chemical formulas of the folic acid antagonist pemetrexed (Figure 1A), representative gammapemetrexed polyglutamate, gammapemetrexed diglutamate (Figure 1B), gammapemetrexed triglutamate (Figures 1C and 1D), gammapemetrexed tetraglutamate (Figures 1E and 1F), gammapemetrexed pentaglutamate (Figures 1G and 1H), gammapemetrexed hexaglutamate (Figures 1I and 1J), gammapemetrexed heptaglutamate (Figures 1K and 1L), and gammapemetrexed octaglutamate (Figures 1M and 1N) are shown. [Figure 2] This study demonstrates the relative potency of liposomal pemetrexed gamma-L hexaglutamate (liposomal aG6) and its enantiomer, liposomal gamma-D hexaglutamate (liposomal aDG6), compared to pemetrexed, after 48 hours of exposure to cancer cell lines SW620 (CRC), HT-29 (colorectal cancer), H1806 (triple-negative breast cancer), OAW28 (ovarian cancer), H292 (NSCLC, adenocarcinoma subtype), and H2342 (NSCLC, adenocarcinoma subtype). [Figure 3] This example shows the dose-response relationship between free pemetrexed L-gammahexaglutamate (gG6), liposomal pemetrexed L-gammahexaglutamate (liposomal gG6), pemetrexed, and folate receptor alpha-targeted antibody (FR1Ab) liposomal pemetrexed L-gammahexaglutamate (liposomal gG6-FR1Ab) in HT-29 (colorectal cancer) cells at 48 hours. [Figure 4] This study demonstrates the effects of free pemetrexed L-gammahexaglutamate (hexa-gG6) and liposomal pemetrexed L-gammahexaglutamate (liposomal hexa-gG6) on the growth of colorectal cancer SW260 cells after 48 hours of exposure to a 256 nM corresponding drug. Untargeted and targeted liposomal pemetrexed hexa-gG6 can enter cells more efficiently than free pemetrexed hexa-gG6 to inhibit the proliferation of colorectal cancer SW260 cells. [Figure 5]This study demonstrates the relative potency of liposomal pemetrexed L-gamma hexaglutamate (liposomal gG6) and its enantiomer, liposomal pemetrexed gamma-D hexaglutamate (liposomal gDG6), compared to pemetrexed, after 48 hours of exposure to cancer cell lines SW620 (CRC), HT-29 (colorectal cancer), H1806 (triple-negative breast cancer), OAW28 (ovarian cancer), H292 (NSCLC, adenocarcinoma subtype), and H2342 (NSCLC, adenocarcinoma subtype). [Figure 6] This study demonstrates the therapeutic effects of liposomal pemetrexed gamma-L hexaglutamate (Lps Hexa gG6), liposomal pemetrexed gamma-D hexaglutamate (Lps Hexa gDG6), and pemetrexed on HCC1806 triple-negative breast cancer cells after 48 hours of exposure. [Figure 7] This study demonstrates the therapeutic effects of liposomal pemetrexed gamma-L hexaglutamate (Lps Hexa gG6) and liposomal pemetrexed gamma-D hexaglutamate (Lps Hexa gDG6) on OAW28 ovarian cancer cells after exposure to pemetrexed over 48 hours. [Figure 8] This study demonstrates the therapeutic effects of liposomal pemetrexed gamma-L hexaglutamate (Lps Hexa gG6), liposomal pemetrexed gamma-D hexaglutamate (Lps Hexa gDG6), and pemetrexed on H292 non-small cell lung cancer cells after 48 hours of exposure. [Figure 9] This study demonstrates the therapeutic effects of liposomal pemetrexed gamma-L hexaglutamate (liposomal gG6), liposomal pemetrexed gamma-D hexaglutamate (liposomal gDG6), and pemetrexed on H292 non-small cell lung cancer cells after 48-hour exposure at various dose levels ranging from 16 to 128 nM. In each tested dose range, the liposomal pemetrexed gG6 formulation showed superior suppression of H292 non-small cell lung cancer cells compared to pemetrexed. [Figure 10]This study demonstrates the therapeutic effects of liposomal pemetrexed gamma-L hexaglutamate (liposomal gG6), liposomal pemetrexed gamma-D hexaglutamate (liposomal gDG6), and pemetrexed on HCC1806 triple-negative breast cancer cells after 48 hours of exposure at various dose levels ranging from 16 to 128 nM. In each tested dose range, the liposomal pemetrexed gG6 formulations were superior to pemetrexed in suppressing HCC1806 triple-negative breast cancer cells. [Figure 11] This study demonstrates the therapeutic effects of liposomal pemetrexed gamma-L hexaglutamate (liposomal gG6), liposomal gamma-D hexaglutamate (liposomal gDG6), and pemetrexed on OAW28 ovarian cancer cells after 48-hour exposure at a range of concentrations. At a dose of 128 nM, pemetrexed appears to be more effective than the liposomal pemetrexed gG6 liposomal formulation, but at doses of 32 nM and 64 nM, the liposomal formulations exhibit superior therapeutic effects compared to pemetrexed, and at 16 nM, the therapeutic effect of liposomal pemetrexed gG6 is similar to that of pemetrexed. [Figure 12] The toxicity of liposomal pemetrexed gamma-L hexaglutamate (liposomal gG6), liposomal pemetrexed gamma-D hexaglutamate (liposomal gDG6), and pemetrexed to differentiated human neutrophils at 64 nM, 128 nM, and 264 nM levels, respectively. The figure shows that liposomal pemetrexed gG6 is significantly less toxic to differentiated human neutrophils than pemetrexed. [Figure 13] This study demonstrates the effects on neutrophils (differentiated from CD34+ cells) after 48 hours of exposure to various dose levels ranging from 16 to 128 nM of liposomal pemetrexed gamma-L hexaglutamate (liposomal gG6), liposomal gamma-D hexaglutamate (liposomal gDG6), and the corresponding pemetrexed agents. [Figure 14]This study demonstrates the effects on AML12 hepatocytes after 48-hour exposure to liposomal pemetrexed gamma-L hexaglutamate (liposomal gG6), liposomal pemetrexed gamma-D hexaglutamate (liposomal gDG6), and the corresponding 16 nM, 32 nM, 64 nM, and 128 nM doses of pemetrexed. Notably, none of the tested dose levels of the liposomal agents appeared to be toxic to AML12 hepatocytes after treatment with liposomal pemetrexed gG6. In contrast, pemetrexed treatment resulted in approximately a 40% reduction in AML12 hepatocyte counts at all doses investigated. [Figure 15] This study demonstrates the effects on CCD841 colon epithelial cells after 48-hour exposure to liposomal pemetrexed gamma-L hexaglutamate (liposomal gG6), liposomal pemetrexed gamma-D hexaglutamate (liposomal gDG6), and the corresponding pemetrexed at concentrations of 16 nM, 32 nM, 64 nM, and 128 nM. At all concentrations tested, pemetrexed resulted in a reduction of approximately 50% or more in CCD841 colon epithelial cell counts, compared to a reduction of approximately 20% or less after treatment with each of the liposomal compositions tested. [Figure 16] The structures of the polyglutamate folate antagonist, cisplatin (CDDP), and two possible gG6-cisplatin complexes are shown. The pH-dependent formation of interchain and / or intrachain coordination between the carboxyl group of the polyglutamate folate antagonist and cisplatin may lead to degradation into separate molecules of gG6 and cisplatin upon encountering acidic lysosomes (pH 3-5) and in the presence of intracellular chloride ions. [Figure 17] Hematological parameters: The effects of liposomal aG6 treatment in mice with weekly administration of 40 mg / kg and 80 mg / kg for 4 weeks are shown on white blood cell (WBC) count, neutrophil count, and platelet count. No significant decrease in mean neutrophils, mean white blood cell count, or mean platelet count was observed. [Figure 18]This study demonstrates the effects of liposomal aG6 therapy on hemoglobin and reticulocyte count index in mice, administered weekly at doses of 40 mg / kg and 80 mg / kg for 4 weeks. Higher dose levels show a minimal decrease in mean hemoglobin concentration, while simultaneously exhibiting a slight increase in mean reticulocyte count index. [Figure 19] This study demonstrates the effects of liposomal aG6 therapy in mice with weekly doses of 40 mg / kg and 80 mg / kg for 4 weeks on liver markers including serum albumin, serum aspartate aminotransferase (AST), and serum alanine aminotransferase (ALT). No significant increase was observed in mean AST or mean ALT levels, and no change in mean albumin levels was observed. [Figure 20] This shows the relative tumor volume of immunodeficient female Nu / J mice (6-8 weeks old) inoculated with NCI-H292 (non-small cell lung cancer) cells and administered intravenously once every 3 weeks at doses of control, pemetrexed, and liposomal aG6 at 167 mg / kg. As these preliminary data show, liposomal aG6 results in reduced tumor control compared to pemetrexed. [Figure 21A-F]Liposomal pemetrexed alpha-L triglutamate was administered over 48 hours to H2342 (NSCLC, adenocarcinoma subtype) (Figure 21A), H292 (NSCLC, adenocarcinoma subtype) (Figure 21B), HT-29 (colon cancer) (Figure 21C), HCC1806 (triple-negative breast cancer) (Figure 21D), MCF7 (ER+ breast cancer) (Figure 21E), and OAW28 (ovarian cancer) (Figure 21F). The dose-response relationships for liposome aG3), liposome pemetrexed alpha-L pentaglutamate (liposome aG5), liposome pemetrexed alpha-L octaglutamate (liposome aG7), and the combination of liposome pemetrexed alpha-L hexaglutamate (aG6) and alpha-L dodecaglutamate (aG12) (liposome aG6 and aG12) are shown. Cell viability was measured by the CellTiter-Glo® (CTG) luminescent cell viability assay, basically as described in Example 1. As shown for all cell lines, the potency of each polyglutamate-oxidized pemetrexed liposome composition well exceeded the potency of the liposome carrier and the empty liposome control. [Modes for carrying out the invention]

[0051] Generally, this disclosure relates to gamma-polyglutamine oxidized folate antimetabolites. The compositions represent an advance in the treatment of existing hyperproliferative diseases such as cancer. Methods for manufacturing, delivering, and using the gamma-polyglutamine oxidized folate antimetabolites are also provided. The gamma-polyglutamine oxidized compositions have applications including, but are not limited to, the treatment or prevention of hyperproliferative diseases such as cancer, immune system disorders including inflammation and autoimmune diseases such as rheumatoid arthritis, and infectious diseases such as HIV, malaria, and schistosomiasis.

[0052] definition Unless otherwise specified, all technical and scientific terms used herein shall have the same meaning as those generally interpreted by those skilled in the art to which this disclosure belongs.

[0053] Whenever an embodiment is described herein with the term “comprising,” other similar embodiments described with the terms “containing,” “consisting of,” and / or “consisting essentially of,” are also provided. However, when used as transitional clauses in the claims, each should be interpreted separately and in the appropriate legal and factual context (for example, in the claims, the transitional clause “comprising” is considered a more open phrase, “consisting of” is considered more exclusive, and “consisting essentially of” is considered intermediate between these).

[0054] As used herein, the singular forms "a," "an," and "the" refer to multiple references unless otherwise specified or unless the context clearly indicates that multiple references are not intended.

[0055] In this specification, the term "and / or" as used in expressions such as "A and / or B" is intended to include both A and B; A or B; A (alone); and B (alone). Similarly, the term "and / or" as used in expressions such as "A, B and / or C" each encompasses the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0056] Headings and subheadings are used for convenience and / or for compliance with official rules only, and are not intended to limit the scope of the subject technology, nor are they referenced in connection with the interpretation of the description of the subject technology. Features described under one heading or subheading of the subject disclosure may be combined with features described under other headings or subheadings in various embodiments. Furthermore, not all features under a single heading or subheading are necessarily used together in some embodiments.

[0057] Unless otherwise specified, the terms “folate antagonist” and “ANTIFOL” are used interchangeably and include salts, acids, and / or free base forms of folate antagonists (e.g., disodium folate antagonist). Compositions containing ANTIFOL salts may further include various cations, e.g., Na + Mg 2+ , K + NH 4+ , and / or Ca 2+ It may include any of the following. In certain embodiments, the salt is a pharmaceutically acceptable salt. The folate antimetabolite contains one L-gamma-glutamyl group and is therefore considered to be monoglutamine-oxidized for the purposes of this disclosure.

[0058] The compounds of the present invention may exist as a mixture of stereoisomers, but it is preferable that they be separated into a single optically active isomer. Such requirements complicate the synthesis of the compounds, and therefore, it is preferable that they contain as few chiral carbon atoms as possible while being compatible with achieving the desired activity.

[0059] However, as previously shown, the cyclopenta[g]quinazoline of the present invention contains at least three chiral carbon atoms. Of these, it is preferable that the chiral carbon atoms at the 6th position of the ring system have a 6S orientation rather than a 6R orientation. The preferred compound (I) described herein is therefore preferred to have such an arrangement at the position of the chiral carbon atoms, and less preferred to be a mixture in which one or both of these chiral carbon atoms are not separated.

[0060] The folate antagonist may be any known or future induced folate or polyglutamine-oxidized folate antagonist. In some embodiments, the folate antagonist may be LV(etoposide), L-leucovorin(L-5-formyltetrahydrofolate); 5-CH3-THF, 5-methyltetrahydrofolate; FA, folate; PteGlu, pteroylglutamate(FA); MTX, methotrexate; 2-dMTX, 2-desamino-MTX; 2-CH3-MTX, 2-desamino-2-methyl-MTX; AMT, aminopterin; 2-dAMT, 2-desamino-AMT; 2-CH3-AMT, 2-desamino-2-methyl-AMT; 10-EdAM, 10-ethyl-10-deazaaminopterin; PT523, N alpha-(4-amino-4-deoxypteroyl)-N Delta-(hemiphthaloyl)-L-ornithine;DDATHF (lometrexol), 5,10-dideaza-5,6,7,8-tetrahydrofolate;5-d(i)H4PteGlu, 5-deaza-5,6,7,8-tetrahydroisofolate;N9-CH3-5-d(i)H4PteGlu, N9-methyl-5-deaza-5,6,7,8-tetrahydroisofolate;5-dPteHCysA,N Alpha-(5-deazapteroyl)-L-homocysteic acid;5-dPteAPBA,N Alpha-(5-deazapteroyl)-DL-2-amino-4-phosphonobutyric acid;5-dPteOrn,N Alpha-(5-deazapteroyl)-L-ornithine;5-dH4PteHCysA,N Alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-L-homocysteic acid; 5-dH4PteAPBA,N Alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-DL-2-amino-4-phosphobutanoic acid; 5-dH4PteOro,N Alpha-(5-dideaza-5,6,7,8-tetrahydropteroyl)-L-ornithine;CB3717,N10-propargyl-5,8-dideazafolate;ICI-198,583,2-desamino-2-methyl-N10-propargyl-5,8-dideazafolate;4-H-ICI-198,583,4-deoxy-ICI-198,583:4-OCH3-ICI-198,583,4-methoxy-ICI-198,583 Glu-to-Val-ICI-198,583;Valine-ICI-198;583;Glu-to-Sub-ICI-198,583,2-aminosuberic acid-ICI-198,583;7-CH3-ICI-198,583,7-methyl-ICI-198,583;ZD1694,N-[5(N-(3,4-dihydro-2-methyl-4-oxoquinazoline-6-yl-methyl)amino)2-thienyl)]-L-glutamic acid ;2-NH2-ZD1694,2-amino-ZD1694;BW1843U89,(S)-2[5-(((1,2-dihydro-3-methyl-1-oxobenzo(f)quinazolin-9-yl)methyl)amino-)-1-oxo-2-isoindolinyl]-glutaric acid;LY231514,N-(4-(2-(2-amino-4,7-dihydro-4-oxo-3H-pyrrolo[2,3-D ]pyrimidine-5-yl)ethyl)-benzoyl]-L-glutamic acid;IAHQ, 5,8-dideazaisofolate;2-dIAHQ, 2-desamino-IAHQ;2-CH3-dIAHQ, 2-desamino-2-methyl-IAHQ;5-d(i)PteGlu, 5-deazaisofolate;N9-CH3-5-d(i)PteGlu, N9-methyl-5-deazaisofolate;N9-CHO- 5-d(i)PteGlu, N9-formyl-5-deazaisofolate; AG337, 3,4-dihydro-2-amino-6-methyl-4-oxo-5-(4-pyridylthio)quanazoline; and AG377, 2,4-diamino-6[N-(4-(phenisulfonyl)benzyl)ethyl)amino]quinazoline; or their stereoisomers, or selected from their stereoisomers.

[0061] In some embodiments, the folate antagonist is a member selected from the following: aminopterin, methotrexate, larcitrexed (also known as TOMUDEX®, ZD1694 (RTX)), previtrexed (BGC9331; also known as ZD9331), pemetrexed (ALIMTA, also known as LY231514), lometrexol (LTX) (5,10-dideazatetrahydrofolate), cyclopenta[g]quinazoline with a dipeptide ligand, CB3717, CB300945 (also known as BGC945), or their stereoisomers such as 6-R, S-BGC945 (ONX-0801), CB300638 (also known as BGC638), and BW1843U89.

[0062] The terms “polyglutamic acid folate antimetabolites,” “polyglutamic acid ANTIFOL,” “ANTIFOL-PG,” and “PANTIFOL” are used herein to mean the same thing and refer to a folate antimetabolite composition (i.e., ANTIFOL-PGn, n≧1) that contains at least one glutamyl group in addition to the glutamyl group in the folate antimetabolite. References to the number of glutamyl groups in γPANTIFOL(ANTIFOL-PG) herein take into account the glutamyl group in the folate antimetabolite. For example, an ANTIFOL-PG composition containing five glutamyl residues in addition to the glutamyl group of ANTIFOL is herein referred to as a hexaglutamic acid folate antimetabolite or folate antimetabolite hexaglutamate. A polyglutamate chain contains an N-terminal glutamyl group and one or more C-terminal glutamyl groups. The N-terminal glutamyl group of a polyglutamate chain does not bond to another glutamyl group via its amino group, but rather to one or more glutamyl groups via its carboxylic acid group. In some embodiments, the N-terminal glutamyl group of a polyglutamic folate antagonist is the glutamyl group of the folate antagonist. The C-terminal glutamyl groups (one or more) of a polyglutamate chain bond to another glutamyl group via their amino groups, but not via their carboxylic acid group.

[0063] The terms “alpha-glutamyl group,” “alpha-glutamate,” “alpha-bond,” and their repetitions mean a glutamyl group containing an alpha-carboxyl group bond when these relate to the bonding of a glutamyl group. In some embodiments, none of the glutamyl groups of the provided polyglutamic oxoxide antimetabolites contain an alpha-bond.

[0064] The terms “gamma-glutamyl group,” “gamma-glutamate,” and “gamma bond” refer to a glutamyl group including a gamma-carboxyl bond when they relate to the bond of a glutamyl group. In some embodiments, the gamma bond is an amide bond between a gamma-carboxyl group of one glutamyl group and a second glutamyl group. The gamma bond may be a bond between a glutamyl group and a glutamyl group in a folate antagonist, or between a glutamyl group and a second glutamyl group, such as a glutamyl group in a polyglutamate chain that is not present in the folate antagonist but is bonded to the folate antagonist. In some embodiments, the gamma bond refers to an amide bond of the glutamyl group of a folate antagonist. Unless otherwise specified or clearly indicated from the context, references to the gamma bond include the gamma bond of the glutamyl group of a folate antagonist. In some embodiments, the gamma-glutamyl group is L-type. In some embodiments, the gamma-glutamyl group is of the D type. As discussed herein, during folate antagonist therapy, the folate antagonist enters the cell and is polyglutamic acid by the enzyme folyl polygamma-glutamate synthase (FPGS), which adds an L-glutamyl group in series to the gamma-carboxyl group of glutamic acid within the L-glutamyl group of the folate antagonist. As a result, the D-gamma-polyglutamic acid folate antagonist composition is not formed inside the cell during folate antagonist therapy.

[0065] The terms “gamma-polyglutamine-oxidized folate antagonist,” “γ-polyglutamine-oxidized folate antagonist,” “γPANTIFOL,” “gamma-polyglutamine-oxidized folate antagonist,” “polyglutamine-oxidized folate antagonist,” “γANTIFOL-PG,” and their repetitions are used herein with the same meaning and refer to a folate antagonist composition comprising at least one gamma-glutamyl group having a gamma-carboxyl group bond in addition to the gamma-glutamyl group in the folate antagonist (e.g., ANTIFOL-PG). n (wherein the formula, n≧1 γ-glutamyl groups). References to the number of glutamyl groups in γPANTIFOL(γANTIFOL-PG) herein take into account the γ-glutamyl groups of the folate antagonist. For example, a γANTIFOL-PG composition containing five γ-glutamyl groups in addition to the glutamyl groups of the folate antagonist may herein be called gamma-hexaglutamate folate antagonist or gamma-folate antagonist hexaglutamate.

[0066] The terms "alpha-glutamyl group," "α-glutamyl group," and "alpha bond" refer to glutamyl groups that include an alpha-carboxyl bond when they relate to the bonding of a glutamyl group.

[0067] As used herein, the term “isolated” means a composition in a form not found in nature. Isolated gamma polyglutamine oxidized compositions include those that have been purified to such an extent that they are no longer in a form found in nature. In some embodiments, the isolated gamma polyglutamine oxidized folate antimetabolites are substantially pure. Isolated compositions are free from or substantially free from naturally incorporated substances such as proteins and other cellular components such as nucleic acids that may be found in nature or in the environment in which they are produced (e.g., cell culture). Gamma polyglutamine oxidized compositions may be formulated with diluents or adjuvants and further isolated for practical purposes—for example, when used as a diagnostic agent or therapy, gamma polyglutamine oxidized compositions are typically mixed with a pharmaceutically acceptable carrier or diluent. In some embodiments, the isolated gamma-polyglutamine oxidation composition (e.g., delivery carriers such as gamma-polyglutamates and gamma-polyglutamates-containing liposomes) contains less than 1% or less than 0.1% of undesirable DNA or protein content. In some embodiments, the gamma-polyglutamate composition (e.g., delivery carriers such as gamma-polyglutamates and gamma-polyglutamates-containing liposomes) is "isolated".

[0068] As used herein, the term “targeting portion” means a molecule that provides enhanced affinity to a selected target, such as a cell, cell type, tissue, organ, region of the body, or compartment, such as a cell, tissue, or compartment of an organ. Targeting portions can include a wide variety of entities. Targeting portions may include native molecules, or recombinant or synthetic molecules. In some embodiments, the targeting portion is an antibody-antigen-binding antibody fragment, a bispecific antibody, or other antibody-based molecule or compound. In some embodiments, the targeting portion is an aptamer, avimer, receptor-binding ligand, nucleic acid, biotin-avidin bond pair, peptide, protein, carbohydrate, lipid, vitamin, toxin, microbial component, hormone, receptor ligand, or any derivative thereof. Other targeting portions are known in the Art and are included herein.

[0069] The term “specific affinity” or “specifically binding” means that a targeting moiety, such as an antibody or antigen-binding antibody fragment, reacts to or binds to an epitope, protein, or target molecule more frequently, more rapidly, for longer periods, with greater affinity, or in some combination of these, than it would to another substance containing a protein unrelated to the target epitope. Due to sequence identity between homologous proteins in different species, specific affinity, in some embodiments, may include a binding agent that recognizes two or more proteins or targets in different species. Similarly, due to homology within specific regions of polypeptide sequences of different proteins, the term “specific affinity” or “specific binding” may include a binding agent that recognizes two or more proteins or targets. In certain embodiments, a targeting moiety that specifically binds to a first target may or may not specifically bind to a second target. Thus, “specific affinity” does not necessarily require (though may include) exclusive binding, e.g., binding to only one target. Therefore, a targeting moiety may, in certain embodiments, specifically bind to two or more targets. In certain embodiments, multiple targets may be combined by the same targeting portion.

[0070] The term "epitope" refers to a portion of an antigen that can be recognized and specifically bound to a targeting portion (i.e., a binding site) such as an antibody. When the antigen is a polypeptide, the epitope can be formed from both continuous and discontinuous amino acids juxtaposed by the protein's tertiary folding. Epitopes formed from continuous amino acids are usually retained during protein denaturation, while epitopes formed by tertiary folding are usually lost during protein denaturation. Epitopes typically contain at least three amino acids, more commonly at least five or eight to ten amino acids, within a distinctive spatial higher-order structure.

[0071] Expressions known in the art such as “binding affinity to target,” “binding to target,” and similar expressions refer to the properties of a targeting moiety that can be directly measured by determining the affinity constant, e.g., the amount of targeting moiety that binds and dissociates at a given antigen concentration. Intermolecular interactions can be characterized using other methods, but are not limited to competitive analysis, equilibrium analysis, and microcalorimetric analysis, and real-time interaction analysis based on surface plasmon resonance interactions (e.g., using a BIACORE® instrument). These methods are well known to those skilled in the art and are described, for example, in Neri et al., Tibtech 14:465-470 (1996) and Jansson et al., J. Biol. Chem. 272:8189-8197 (1997).

[0072] The term “delivery carrier” typically refers to any composition that assists, promotes, or facilitates the entry of gamma-polyglutamine oxidase antagonists into cells. Such delivery carriers are known in the art and are not limited to, but include liposomes, lipospheres, polymers (e.g., polymer complexes), peptides, proteins such as antibodies (e.g., immune complexes such as antibody-drug conjugates (ADCs) and antigen-binding antibody fragments and their derivatives), cellular components, cyclic oligosaccharides (e.g., cyclodextrins), micelles, microparticles (e.g., microspheres), nanoparticles (e.g., lipid nanoparticles, biodegradable nanoparticles, and core-shell nanoparticles), hydrogels, lipoprotein particles, viral sequences, viral substances, or lipid or liposomal formulations, and combinations thereof. The delivery carrier can be directly or indirectly bound to the targeting moiety. In some examples, the targeting moiety is selected from polymers, proteins, peptides, monoclonal antibodies, or fatty acid lipids.

[0073] "Subject" means humans or, but not limited to, dogs, cats, horses, goats, and primates, including vertebrate mammals such as monkeys. Accordingly, the present invention can also be used to treat diseases or conditions in non-human subjects. For example, cancer is one of the leading causes of death in companion animals (e.g., cats and dogs). In some embodiments of the present invention, the subject is human. In this disclosure, the terms "subject" and "patient" are used interchangeably and have the same meaning. In general, it is preferable to use the maximum dose, i.e., the maximum safe dose according to sound medical judgment.

[0074] As used herein, “effective dose” means a dose of the drug sufficient to produce the medically desired result. The effective dose may vary depending on the desired outcome, the specific condition being treated or prevented, the age and health status of the person being treated, the severity of the condition, the duration of treatment, the nature of any concurrent or adjunctive therapies, the specific route of administration, and similar factors within the scope of the knowledge and professional opinion of a healthcare practitioner. The “effective dose” can be determined experimentally and routinely in relation to the stated purpose. In the case of cancer, an effective dose of the drug may reduce the number of cancer cells; reduce the size of the tumor; inhibit (i.e., slow to some extent, preferably stop) the invasion of cancer cells into surrounding organs; inhibit (i.e., slow to some extent, preferably stop) the metastasis of the tumor; inhibit (i.e., slow to some extent, preferably stop) the growth of the tumor; and / or alleviate to some extent one or more of the symptoms associated with the disorder. Depending on the extent to which the drug can prevent and / or kill existing cancer cells, the drug may be inhibitory and / or cytotoxic. For cancer therapy, in vivo efficacy can be measured, for example, by evaluating survival time, progression-free survival (PFS) time, response rate (RR), response duration, and / or quality of life.

[0075] The terms “hyperproliferative disorder,” “proliferative disorder,” and “proliferative disorder” are used herein to mean the same thing and relate to the undesirable or uncontrolled proliferation of unwanted, excessive or abnormal cells, such as neoplastic or hyperplastic proliferation, whether in vitro or in vivo. In some embodiments, proliferative disorder is cancer or neoplastic disease (including benign or malignant) and / or any tumor metastasis, regardless of the location of the cancer, tumor and / or tumor metastasis. In some embodiments, proliferative disorder is a benign or malignant tumor. In some embodiments, proliferative disorder is a non-cancerous disease. In some embodiments, proliferative disorder is an overgrowth condition such as hyperplasia, fibrosis (in particular pulmonary, but also other types of fibrosis such as renal fibrosis), angiogenesis, psoriasis, atherosclerosis and smooth muscle proliferation in blood vessels such as stenosis or restenosis after angiogenesis.

[0076] The terms “cancer,” “tumor,” or “malignant tumor” are used synonymously to mean any of many diseases characterized by uncontrolled, abnormal proliferation of cells, localized or metastatic spread of infected cells to other parts of the body via the bloodstream and lymphatic system, and numerous distinctive structural and / or molecular features. As used herein, “tumor” means all neoplastic cell growth and proliferation, whether malignant or benign, as well as all precancerous and cancerous cells and tissues. “Malignant tumor,” or “malignant cell,” is understood to be a cell that has specific structural characteristics, lacks differentiation, and is capable of invasion and metastasis. Cancers that can be treated with the γPANTIFOL compositions provided herein include, but are not limited to, non-hematological malignancies such as lung cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, head and neck cancer, gastric cancer, gastrointestinal cancer, colorectal cancer, esophageal cancer, cervical cancer, liver cancer, kidney cancer, bile duct cancer, gallbladder cancer, bladder cancer, sarcomas (e.g., osteosarcoma), brain cancer, central nervous system cancer, and melanoma; and hematological malignancies such as leukemia, lymphoma and other B-cell malignancies, myeloma and other plasmacytotic dysplasia or cachexia. In some embodiments, cancer is selected from breast cancer, advanced head and neck cancer, lung cancer, gastric cancer, osteosarcoma, non-Hodgkin lymphoma (NHL), acute lymphoblastic leukemia (ALL), mycosis fungoides (cutaneous T-cell lymphoma), choriocarcinoma, chorioadenoma, non-leukemic meningeal carcinomatosis, soft tissue sarcoma (tendonoid, invasive fibromatosis), bladder cancer, and central nervous system (CNS) cancer. Other types of cancer and tumors that can be treated with γPANTIFOL compositions are described herein or known in the art. The term “metastasis” means the spread or dissemination to other sites, locations, regions or organs or tissue systems within a tumor, cancer, or tumor subject, where the sites, locations, regions or organs or tissue systems within the subject are distinct from the primary tumor, cancer, or neoplasm. The terms “cancer,” “cancerous,” “proliferative disorder,” “proliferative disorder,” and “tumor” are not mutually exclusive when referenced herein.

[0077] Terms such as “to treat,” “to cure,” or “to treat” mean both (a) therapeutic means that cure, slow, reduce, and / or halt the progression of the symptoms of a diagnosed condition or disorder, and (b) preventive or protective means that prevent and / or delay the onset of a targeted disease or condition. Accordingly, subjects requiring treatment include subjects who already have the cancer, disorder or disease, subjects at risk of developing the cancer or condition, and subjects for whom infection or condition should be prevented. Subjects are identified using well-known medical and diagnostic techniques as “at risk of having” cancer, infectious disease, immune system disorder, hyperproliferative disorder, or another disease or disorder as referred herein. In certain embodiments, if a subject exhibits overall, partial, or temporary remission or elimination of symptoms associated with a disease or condition (e.g., cancer, inflammation, and rheumatoid arthritis), the subject is “treated” by the methods provided herein. In certain embodiments, the terms “treating,” “treatment,” or “treat” mean improvement of at least one measurable physical parameter of a proliferative disorder, such as tumor growth, which may not be identifiable by the patient. In other embodiments, the terms “treating,” “treatment,” or “treat” mean suppression of the progression of a proliferative disorder, for example, physically by stabilizing identifiable symptoms, or physiologically, for example, by stabilizing physical parameters, or both. In other embodiments, the terms “treating,” “treatment,” or “treat” mean reduction or stabilization of size, tumor cell proliferation or survival, or cancer cell number. Treatment may involve using the γPANTIFOL composition alone or in combination with additional therapeutic agents.

[0078] The terms “subject,” “patient,” and “animal” are used synonymously and mean human patients and mammals such as non-human primates, as well as laboratory animals such as rabbits, rats, and mice, and other animals. Animals include all vertebrates, mammals and non-mammals such as chickens, amphibians, and reptiles. As used herein, “mammal” means any member of the class Mammalia, including, but not limited to, humans and non-human primates, such as chimpanzees and other apes and monkey species; domestic animals such as cattle, sheep, pigs, goats, and horses; domestic mammals such as dogs and cats; laboratory animals such as rodents such as mice, rats, and guinea pigs, and other members of the class Mammalia known in the art. In certain embodiments, the patient is human.

[0079] As used herein, “treatment of proliferative disorders” includes maintaining or reducing the size of the tumor of the subject of the proliferative disorder, inducing tumor reduction (partial or complete), suppressing tumor growth, and / or extending lifespan. In one embodiment, the proliferative disorder is a solid tumor. Such tumors include, for example, lung cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, head and neck cancer, gastric cancer, gastrointestinal cancer, colorectal cancer, esophageal cancer, cervical cancer, liver cancer, kidney cancer, bile duct cancer, gallbladder cancer, bladder cancer, sarcoma (e.g., osteosarcoma), brain cancer, central nervous system cancer, and melanoma. In one embodiment, the proliferative disorder is a hematological malignancy. Such hematological malignancies include, for example, leukemia, lymphoma and other B-cell malignancies, myeloma and other plasmacytotic dysplasia or cachexia.

[0080] As used herein, the term “autoimmune disease” is defined as a disorder resulting from an autoimmune reaction. Autoimmune diseases are the result of an inappropriate and excessive reaction to autoantigens. Examples of autoimmune diseases include, but are not limited to, Addison's disease, alopecia areata, ankylosing spondylitis, autoimmune hepatitis, autoimmune mumps, Crohn's disease, diabetes mellitus (type 1), dystrophic epidermolysis bullosa, epididymitis, glomerulonephritis, Graves' disease, Guillain-Barré syndrome, Hashimoto's disease, hemolytic anemia, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, pemphigus vulgaris, psoriasis, rheumatic fever, rheumatoid arthritis, sarcoidosis, scleroderma, Sjögren's syndrome, spondyloarthritis, thyroiditis, vasculitis, vitiligo, myxedema, pernicious anemia, and ulcerative colitis.

[0081] The terms "inflammation" and "inflammatory disease" are used interchangeably and refer to diseases or disorders characterized by or caused by inflammation. "Inflammation" refers to a localized response to cellular damage characterized by capillary dilation, leukocyte infiltration, redness, heat, and pain, which play a role in initiating the removal of harmful drugs and damaged tissue. Sites of inflammation include the lungs, pleura, tendons, lymph nodes or glands, uvula, vagina, brain, spinal cord, nasal and pharyngeal mucosa, muscles, skin, bone or bone tissue, joints, bladder, retina, cervix, canals of the eyes, intestines, vertebrae, rectum, anus, bursae, hair follicles, etc. Such inflammatory diseases include, but are not limited to, inflammatory bowel disease, rheumatoid arthritis (e.g., rheumatoid arthritis), other arthritis (e.g., acute arthritis, acute gouty arthritis, bacterial arthritis, chronic inflammatory arthritis, osteoarthritis, infectious arthritis, juvenile arthritis, fungal arthritis, neuropathic arthritis, polyarthritis, proliferative arthritis, psoriatic arthritis, sexually transmitted arthritis, viral arthritis), connective tissue inflammation, pelvic inflammatory disease, acne, psoriasis, actinomycosis, dysentery, biliary cirrhosis, Lyme disease, heat rash, Stevens-Johnson syndrome, mumps, pemphigus vulgaris, and blastomycosis. Inflammatory bowel disease is a chronic inflammatory disease of the gastrointestinal tract that includes, but is not limited to, Crohn's disease, ulcerative colitis, and unclassified colitis. Rheumatoid arthritis is a chronic inflammatory disease of the joints, usually polyarticular, characterized by inflammatory changes in the synovial membrane and joint structures, as well as muscle spasms and bone roughening.

[0082] As used herein, the term “therapeutic agent” means a drug or its derivatives or prodrugs that interact with overproliferating cells, such as cancer cells or immune cells, thereby reducing the proliferative state of the cells and / or killing them. Examples of therapeutic agents include, but are not limited to, chemotherapeutic agents, cytotoxic agents, platinum-based drugs (e.g., cisplatin, carboplatin, oxaliplatin), taxanes (e.g., Taxol®), etoposide, alkylating agents (e.g., cyclophosphamide, ifosfamide), antimetabolites (e.g., folic acid antimetabolites (ANTIFOL)), 5-fluorouracil, gemcitabine, or its derivatives), antitumor antibiotics (e.g., mitomycin, doxorubicin), and plant-derived antitumor agents (e.g., vincristine, vindesine, Taxol). Such agents include, but are not limited to, the anticancer agents trimethrexate, temozolomide, larcitrexed, S-(4-nitrobenzyl)-6-thioinosine (NBMPR), 6-benziguanidine (6-BG), bis-chloronitrosourea (BCNU), and camptothecin™, or any therapeutic derivative thereof. Further examples of therapeutic agents suitable for use by the methods of this disclosure include, but are not limited to, anti-restenotic agents, proliferative or antiproliferative agents, anti-inflammatory agents, antineoplastic agents, antimitotic agents, antiplatelet agents, anticoagulants, antifibrin agents, antithrombin agents, cell proliferation inhibitors, antibiotics and other anti-infective agents, anti-enzyme agents, antimetabolite agents, angiogenic agents, cytoprotective agents, angiotensin-converting enzyme (ACE) inhibitors, angiotensin II receptor antagonists, and / or cardioprotective agents. "Therapeutic agent" also means the salts, acids, and free base forms of the above agents.

[0083] As used herein, the term “chemotherapeutic agent” means any agent that causes the death of cancer cells or inhibits the growth or spread of cancer cells, when used in connection with cancer therapy. Examples of such chemotherapeutic agents include alkylating agents, antibiotics, antimetabolites, plant-derived drugs, and hormones. In some embodiments, the chemotherapeutic agent is cisplatin. In some embodiments, the chemotherapeutic agent is carboplatin. In some embodiments, the chemotherapeutic agent is oxaliplatin. In other embodiments, the chemotherapeutic agent is gemcitabine. In other embodiments, the chemotherapeutic agent is doxorubicin.

[0084] As used herein, the term “antometabolite” means an antitumor agent that inhibits the utilization of a metabolite or its prodrug. Examples of antimetabolites include folate antimetabolites, pemetrexed, 5-fluorouracil, 5-fluorouracil prodrugs such as capecitabine, 5-fluorodeoxyuridine monophosphate, cytarabine, nerarabine prodrugs, 5-azacitidine, gemcitabine, mercaptopurine, thioguanine, azathioprine, adenosine, pentostatin, erythrohydroxynonyladenine, and cladribine. Nucleoside analogs, including purines or pyrimidine analogs, are useful antimetabolites for carrying out the methods disclosed herein. In some embodiments, the gamma-polyglutamine oxidase antimetaboliticant composition is used in combination with antimetaboliticant selected from fluoropyrimidine, 5-fluorouracil, 5-fluoro-2'-deoxycytidine, cytarabine, gemcitabine, troxacitabine, decitabine, azacitidine, pseudoisocytidine, zebralin, ancitabine, fazarabine, 6-azacitidine, capecitabine, N4-octadecylcytarabine, elaidate cytarabine, fludarabine, cladribine, clofarabine, nerarabine, folodesine, and pentostatin, or derivatives thereof. In one example, the nucleoside analog is a substrate of a nucleoside deaminase, which is adenosine deaminase or cytidine deaminase. In some examples, the nucleoside analog is selected from fludarabine, cytarabine, gemcitabine, decitabine, and azacitidine or their derivatives. In certain embodiments, the antimetabolite is 5-fluorouracil.

[0085] As used herein, “taxane” is an anticancer agent that interferes with or disrupts microtubule stability, formation, and / or function. Taxanes include paclitaxel and docetaxel and their derivatives, the derivatives of which function on microtubules in the same mode of action as the taxanes from which they are derived. In certain embodiments, the taxane is paclitaxel or docetaxel, or a pharmaceutically acceptable salt, acid, or derivative of paclitaxel or docetaxel. In certain embodiments, the taxane is paclitaxel (Taxol®), docetaxel (Taxotere®), albumin-conjugated paclitaxel (nab-paclitaxel; Abraxane®), DHA-paclitaxel, or PG-paclitaxel.

[0086] The terms "pharmaceutically acceptable carrier" and "pharmaceutically acceptable carrier" refer to components in a pharmaceutical preparation other than the active ingredient that are non-toxic to the target. Examples of pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives. Examples of pharmaceutically acceptable carriers include one or more compatible solid or liquid fillers, diluents, or encapsulating materials suitable for administration to humans or other subjects.

[0087] This disclosure generally relates to gamma polyglutamine oxoxide antimetabolites (γANTIFOL) compositions, and to methods for manufacturing and using compositions for treating diseases including hyperproliferative diseases such as cancer, immune system disorders such as rheumatoid arthritis, and infectious diseases such as HIV, malaria, and schistosomiasis.

[0088] In some embodiments, this disclosure provides the following: [1] A composition comprising a gamma-polyglutamine oxoxide antimetabolitic agent; [2] A composition according to item [1], wherein the folate antagonist is selected from pyritrexime, pralatrexate, AG2034, GW1843, and LY309887 and / or their stereoisomers; [3] A composition according to item [1], wherein the folate antagonist is selected from PMX, MTX, RTX, and LTX, or their stereoisomers; [4] A composition described in any of items [1] to [3], wherein the folate antagonist is selected from the following: LV (etoposide), L-leucovorin (L-5-formyltetrahydrofolate); 5-CH3-THF, 5-methyltetrahydrofolate; FA, folate; PteGlu, pteroylglutamate (FA); MTX, methotrexate; 2-dMTX, 2-desamino-MTX; 2-CH3-MTX, 2-desamino-2-methyl-MTX; AMT, aminopterin; 2-dAMT, 2-desamino-AMT; 2-CH3-AMT, 2-desamino-2-methyl-AMT; 10-EdAM, 10-ethyl-10-deazaaminopterin; PT523, N alpha-(4-amino-4-deoxypteroyl)-N Delta-(hemiphthaloyl)-L-ornithine;DDATHF (lometrexol), 5,10-dideaza-5,6,7,8-tetrahydrofolate;5-d(i)H4PteGlu, 5-deaza-5,6,7,8-tetrahydroisofolate;N9-CH3-5-d(i)H4PteGlu, N9-methyl-5-deaza-5,6,7,8-tetrahydroisofolate;5-dPteHCysA,N Alpha-(5-deazapteroyl)-L-homocysteic acid;5-dPteAPBA,N Alpha-(5-deazapteroyl)-DL-2-amino-4-phosphonobutyric acid;5-dPteOrn,N Alpha-(5-deazapteroyl)-L-ornithine;5-dH4PteHCysA,N Alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-L-homocysteic acid; 5-dH4PteAPBA,N Alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-DL-2-amino-4-phosphobutanoic acid; 5-dH4PteOro,N Alpha-(5-dideaza-5,6,7,8-tetrahydropteroyl)-L-ornithine;CB3717, N10-propargyl-5,8-dideazafolate;ICI-198,583, 2-desamino-2-methyl-N10-propargyl-5,8-dideazafolate;4-H-ICI-198,583, 4-deoxy-ICI-198,583:4-OCH3-ICI-198,583, 4-methoxy-ICI-198,583 Glu-to-Val-ICI-198,583;Valine-ICI-198;583;Glu-to-Sub-ICI-198,583,2-aminosuberic acid-ICI-198,583;7-CH3-ICI-198,583,7-methyl-ICI-198,583;ZD1694,N-[5(N-(3,4-dihydro-2-methyl-4-oxoquinazoline-6-yl-methyl)amino)2-thienyl)]-L-glutamic acid;2-NH2 -ZD1694, 2-amino-ZD1694;BW1843U89, (S)-2[5-(((1,2-dihydro-3-methyl-1-oxobenzo(f)quinazolin-9-yl)methyl)amino-)-1-oxo-2-isoindolinyl]-glutaric acid;LY231514, N-(4-(2-(2-amino-4,7-dihydro-4-oxo-3H-pyrrolo[ 2,3-D]pyrimidine-5-yl)ethyl)-benzoyl]-L-glutamic acid; IAHQ, 5,8-dideazaisofolate; 2-dIAHQ, 2-desamino-IAHQ; 2-CH3-dIAHQ, 2-desamino-2-methyl-IAHQ; 5-d(i)PteGlu, 5-deazaisofolate; N9-CH3-5-d(i)PteGlu, N9-methyl-5-deazaisofolate; N9-CHO-5-d(i)PteGlu, N9-formyl-5-deazaisofolate; AG337, 3,4-dihydro-2-amino-6-methyl-4-oxo-5-(4-pyridylthio)quanazoline; and 2,4-diamino-6[N-(4-(phenisulfonyl)benzyl)ethyl)amino]quinazoline; or their stereoisomers. ; A composition of [5][1] wherein the folate antagonist is selected from methotrexate, larcitrexed, previtrexed, pemetrexed, lometrexol (LTX; 5,10-dideazatetrahydrofolate), cyclopenta[g]quinazoline having a dipeptide ligand, CB3717, CB300945, or stereoisomers thereof such as 6-R, S-BGC945 (ONX-0801), CB300638, and BW1843U89; [6] A composition according to any of items [1] to [5], wherein the gamma polyglutamine oxidized folate antimetabolitic agent contains 4, 5, 2 to 10, 4 to 6, or more than 5 glutamyl groups: [7] A composition according to any of items [1] to [6], wherein the gamma polyglutamine oxoxide antimetabolitic agent is (a) Is it a gamma-tetraglutamine oxoxide antimetabolitic agent? (b) gammapentaglobulin oxoxide antimetabolites, or (c) It is a gamma-hexaglutamine oxidase folate antagonist. composition; [8] A composition according to any of items [1] to [7], wherein the gamma-polyglutamine oxidized folate antimetabolitic agent comprises 1 to 10 glutamyl groups having gamma-carboxyl group bonds; [9] A composition described in any of items [1] to [8], (a) At least two glutamyl groups of the gamma polyglutamine oxoxide antimetabolites are in the L form; (b) Each glutamyl group in the gamma polyglutamine oxoxide antimetabolites is L-type; (c) At least one glutamyl group of the gamma polyglutamine oxoxide antimetabolite is of the D type; (d) The glutamyl groups of each gamma polyglutamate oxidase folate antagonist, other than the glutamyl group of the folate antagonist, are of type D; or (e) At least two glutamyl groups of the gamma polyglutamine oxoxide antimetabolites are L-type and at least one glutamyl group is D-type. Composition:

[10] A composition according to any of items [1] to [9], wherein the polyglutamic acid is linear;

[11] A composition according to any of items [1] to [9], wherein the polyglutamic acid is a branched chain;

[12] Liposome composition containing a gamma polyglutamine oxidase folate antimetabolitic agent as described in any of items [1] to

[11] (Lp-γPANTIFOL);

[13] An Lp-γPANTIFOL composition as described in item

[12] , wherein the polyglutamic acid oxidase antagonist is selected from the following: (a) AG2034, pyritrexime, pralatrexate, GW1843, folate antagonists, and LY309887; or (b) PMX, MTX, RTX and LTX, and their stereoisomers;

[14] Lp-γPANTIFOL compositions according to item

[12] or

[13] , wherein the polyglutamic acid oxidase antagonist is selected from the following: LV (etoposide), L-leucovorin (L-5-formyltetrahydrofolate); 5-CH3-THF, 5-methyltetrahydrofolate; FA, folic acid; PteGlu, pteroylglutamate (FA); MTX, methotrexate; 2-dMTX, 2-desamino-MTX; 2-CH3-MTX, 2-desamino-2-methyl-MTX; AMT, aminopterin; 2-dAMT, 2-desamino-AMT; 2-CH3-AMT, 2-desamino-2-methyl-AMT; 10-EdAM, 10-ethyl-10-deazaaminopterin; PT523, N Alpha-(4-amino-4-deoxypterol)-N delta-(hemiphthaloyl)-L-ornithine;DDATHF (lometrexol), 5,10-dideaza-5,6,7,8-tetrahydrofolate;5-d(i)H4PteGlu, 5-deaza-5,6,7,8-tetrahydroisofolate;N9-CH3-5-d(i)H4PteGlu, N9-methyl-5-deaza-5,6,7,8-tetrahydroisofolate;5-dPteHCysA,N alpha-(5-deazapteroyl)-L-homocysteinic acid;5-dPteAPBA,N alpha-(5-deazapteroyl)-DL-2-amino-4-phosphonobutyric acid;5-dPteOrn,N Alpha-(5-deazapteroyl)-L-ornithine; 5-dH4PteHCysA,N Alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-L-homocysteic acid; 5-dH4PteAPBA,N Alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-DL-2-amino-4-phosphobutanoic acid; 5-dH4PteOro,N Alpha-(5-dideaza-5,6,7,8-tetrahydropteroyl)-L-ornithine; CB3717,N10-propargyl-5,8-dideazafolate; ICI-198,583,2-desamino-2-methyl-N10-propargyl-5,8-dideazafolate;4-H-ICI-198,583, 4-deoxy-ICI-198,583:4-OCH3-ICI-198,583, 4-methoxy-ICI-198,583 Glu-to-Val-ICI-198,583;Valine-ICI-198;583;Glu-to-Sub-ICI-198,583, 2-aminosuberic acid-ICI-198,583;7-CH3-ICI-198,583,7-methyl-ICI-198,583;ZD1694, N-[5(N-(3,4-dihydro-2-methyl-4-oxoquinazoline-6-yl-methyl (amino)2-thienyl)-L-glutamic acid; 2-NH2-ZD1694, 2-amino-ZD1694; BW1843U89, (S)-2[5-(((1,2-dihydro-3-methyl-1-oxobenzo(f)quinazolin-9-yl)methyl)amino-)-1-oxo-2-isoindlinyl]-glutaric acid; LY231514, N-(4-(2-(2-amino-4,7-dihydroxy Dro-4-oxo-3H-pyrrolo[2,3-D]pyrimidine-5-yl)ethyl)-benzoyl]-L-glutamic acid;IAHQ, 5,8-dideazaisofolate;2-dIAHQ, 2-desamino-IAHQ;2-CH3-dIAHQ, 2-desamino-2-methyl-IAHQ;5-d(i)PteGlu, 5-deazaisofolate;N9-CH3-5-d(i)PteGlu,N9- Methyl-5-deazaisofolate; N9-CHO-5-d(i)PteGlu, N9-formyl-5-deazaisofolate; AG337, 3,4-dihydro-2-amino-6-methyl-4-oxo-5-(4-pyridylthio)quanazoline; and AG377, 2,4-diamino-6[N-(4-(phenysulfonyl)benzyl)ethyl)amino]quinazoline; or their stereoisomers;

[15] Lp-γPANTIFOL compositions according to any of items

[12] to

[14] , wherein the folate antagonist is selected from methotrexate, larcitrexed, previtrexed, pemetrexed, lometrexol (LMX; 5,10-dideazatetrahydrofolate), cyclopenta[g]quinazoline having a dipeptide ligand, CB3717, CB300945, or stereoisomers thereof such as 6-R, S-BGC945 (ONX-0801), CB300638, and BW1843U89;

[16] An Lp-γPANTIFOL composition according to any of items

[12] to

[15] , wherein the liposomes contain a gamma polyglutamic oxidase folate antimetabolitic agent comprising 4, 5, 2 to 10, 4 to 6, or more than 5 gamma glutamyl groups;

[17] An Lp-γPANTIFOL composition according to any of items

[12] to

[16] , wherein the liposome contains a gammatetraglutamine oxidase folate antimetabolitic agent;

[18] An Lp-γPANTIFOL composition according to any of items

[12] to

[16] , wherein the liposome comprises a gammapentaglobulin oxidase folate antimetabolitic agent;

[19] An Lp-γPANTIFOL composition according to any of items

[12] to

[16] , wherein the liposome contains a gamma-hexaglutamine oxidase folate antimetabolitic antagonist;

[20] An Lp-γPANTIFOL composition according to any of items

[12] to

[19] , wherein the gamma-polyglutamine oxidized folate antimetabolitic agent comprises 1 to 10 glutamyl groups having gamma-carboxyl group bonds;

[21] An Lp-γPANTIFOL composition according to any of items

[12] to

[20] , (a) At least two glutamyl groups of the gamma polyglutamine oxoxide antimetabolites are in the L form; (b) Each glutamyl group in the gamma polyglutamine oxoxide antimetabolites is L-type; (c) At least one glutamyl group of the gamma polyglutamine oxoxide antimetabolite is of the D type; (d) The glutamyl groups of each gamma polyglutamate oxidase folate antagonist, other than the glutamyl group of the folate antagonist, are of type D; or (e) At least two glutamyl groups of the gamma polyglutamine oxoxide antimetabolites are L-type and at least one glutamyl group is D-type. Lp-γPANTIFOL composition:

[22] An Lp-γPANTIFOL composition according to any of items

[12] to

[21] , which: (a) At least two glutamyl groups of the gamma polyglutamine oxoxide antimetabolites are in the L form; (b) Each glutamyl group in the gamma polyglutamine oxoxide antimetabolites is L-type; (c) At least one glutamyl group of the gamma polyglutamine oxoxide antimetabolite is of the D type; (d) The glutamyl groups of each gamma polyglutamate oxidase folate antagonist, other than the glutamyl group of the folate antagonist, are of type D; or (e) At least two glutamyl groups of the gamma polyglutamine oxoxide antimetabolites are L-type and at least one glutamyl group is D-type. Lp-γPANTIFOL composition:

[23] An Lp-γPANTIFOL composition according to any of items

[12] to

[22] , wherein the liposomes are pegylated (PLp-γPANTIFOL);

[24] An Lp-γPANTIFOL composition according to any of items

[12] to

[22] , wherein the liposomes are not pegylated;

[25] An Lp-γPANTIFOL composition according to any of items

[12] to

[24] , wherein the liposomes have a diameter in the range of 20 nm to 200 nm;

[26] An Lp-γPANTIFOL composition according to any of items

[12] to

[25] , wherein the liposomes have a diameter in the range of 80 nm to 120 nm;

[27] An Lp-γPANTIFOL composition according to any of items

[12] to

[26] , wherein the liposomes are formed from liposome components;

[28] An Lp-γPANTIFOL composition as described in item

[27] , wherein the liposome component comprises at least one anionic lipid and a neutral lipid;

[29] Lp-γPANTIFOL compositions according to item

[27] or

[28] , wherein the liposome component comprises at least one selected from DSPE; DSPE-PEG; DSPE-PEG-maleimide; HSPC; HSPC-PEG; cholesterol; cholesterol-PEG; and cholesterol-maleimide;

[30] Lp-γPANTIFOL compositions according to any of items

[27] to

[29] , wherein the liposome component comprises at least one selected from the following: Lp-γPANTIFOL compositions; DSPE; DSPE-PEG; DSPE-PEG-FITC; DSPE-PEG-maleimide; cholesterol; and HSPC;

[31] An Lp-γPANTIFOL composition according to any of items

[27] to

[30] , wherein one or more liposome components further comprise a steric stabilizer;

[32] Lp-γPANTIFOL compositions according to item

[31] , wherein the steric stabilizer is polyethylene glycol (PEG); poly-L-lysine (PLL); monosialoganglioside (GM1); poly(vinylpyrrolidone) (PVP); poly(acrylamide) (PAA); poly(2-methyl-2-oxazoline); poly(2-ethyl-2-oxazoline); phosphatidyl polyglycerol; poly[N-(2-hydroxypropyl)methacrylamide]; amphiphilic poly-N-vinylpyrrolidone; L-amino acid-based polymer; oligoglycerin, polyethylene glycol and polypropylene oxide-containing copolymer, poloxamer 188, and polyvinyl alcohol;

[33] An Lp-γPANTIFOL composition as described in item

[32] , wherein the steric stabilizer is PEG, and the PEG has a number-average molecular weight (Mn) of 200 to 5000 daltons;

[34] An Lp-γPANTIFOL composition according to any of items

[12] to

[33] , wherein the liposomes are anionic or neutral;

[35] An Lp-γPANTIFOL composition according to any of items

[12] to

[33] , wherein the liposomes have a zeta potential of zero or less;

[36] An Lp-γPANTIFOL composition according to any of items

[12] to

[33] , wherein the liposomes have a zeta potential of 0 to -150 mV;

[37] An Lp-γPANTIFOL composition according to any of items

[12] to

[33] , wherein the liposomes have a zeta potential of -30 to -50 mV;

[38] An Lp-γPANTIFOL composition according to any of items

[12] to

[33] , wherein the liposomes are cationic;

[39] An Lp-γPANTIFOL composition according to any of items

[12] to

[38] , wherein the liposome has an internal space containing a gamma polyglutamine oxidized folate antimetabolitic antagonist and an aqueous pharmaceutically acceptable carrier;

[40] Lp-γPANTIFOL compositions as described in item

[39] , wherein the pharmaceutically acceptable carrier comprises an isotonic agent such as dextrose, mannitol, glycerol, potassium chloride, or sodium chloride in a concentration greater than 1%;

[41] An Lp-γPANTIFOL composition as described in item

[39] , wherein the aqueous pharmaceutically acceptable carrier is trehalose;

[42] Lp-γPANTIFOL compositions as described in item

[41] , wherein the pharmaceutically acceptable carrier is 1% to 50% trehalose;

[43] An Lp-γPANTIFOL composition according to any of items

[39] to

[42] , wherein the pharmaceutically acceptable carrier comprises a 1% to 50% dextrose solution;

[44] An Lp-γPANTIFOL composition according to any of items

[39] to

[43] , wherein the internal space of the liposome contains 5% dextrose suspended in HEPES buffer solution;

[45] An Lp-γPANTIFOL composition according to any of items

[39] to

[44] , wherein the pharmaceutically acceptable carrier comprises a buffer such as HEPES buffered saline (HBS) or an analogue with a concentration of 1 to 200 mM and a pH of 2 to 8;

[46] An Lp-γPANTIFOL composition according to any of items

[39] to

[45] , wherein the pharmaceutically acceptable carrier comprises sodium acetate and calcium acetate in a total concentration of 50 mM to 500 mM;

[47] An Lp-γPANTIFOL composition according to any of items

[12] to

[46] , wherein the internal space of the liposomes has a pH of 5 to 8 or a pH of 6 to 7, or any range in between;

[48] ​​An Lp-γPANTIFOL composition according to any of items

[12] to

[47] , wherein the liposomes contain less than 500,000 or less than 200,000 gamma polyglutamine oxidized folate antimetabolitic antagonist molecules;

[49] An Lp-γPANTIFOL composition according to any of items

[12] to

[48] , wherein the liposomes contain 10 to 100,000 or any range in between gamma polyglutamine oxidized folate antimetabolitic antagonist molecules;

[50] An Lp-γPANTIFOL composition according to any of items

[12] to

[49] , further comprising a targeting moiety, wherein the targeting moiety has specific affinity for a surface antigen on a target cell of interest;

[51] An Lp-γPANTIFOL composition as described in item

[50] , wherein the targeting portion is bound to one or both of the PEG and outer surface of the liposome, and optionally, the targeting portion is covalently bound to one or both of the PEG and outer surface of the liposome;

[52] An Lp-γPANTIFOL composition according to item

[50] or

[51] , wherein the targeting portion is a polypeptide;

[53] An Lp-γPANTIFOL composition according to any of items

[50] to

[52] , wherein the targeting portion is an antibody or an antigen-binding fragment of an antibody;

[54] An Lp-γPANTIFOL composition according to any of items

[50] to

[53] , wherein the targeting portion is measured by BIACORE® analysis to be 0.5 x 10 -10 ~10x10 -6Lp-γPANTIFOL composition that binds to surface antigens with an equilibrium dissociation constant (Kd) in the range of [value];

[55] An Lp-γPANTIFOL composition according to any of items

[50] to

[54] , wherein the targeting portion specifically binds to 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);

[56] An Lp-γPANTIFOL composition according to any of items

[50] to

[55] , wherein the targeting portion comprises one or more selected from the group consisting of antibodies, humanized antibodies, antigen-binding fragments of antibodies, single-chain antibodies, single-domain antibodies, bispecific antibodies, synthetic antibodies, pegylated antibodies, and multimeric antibodies;

[57] An Lp-γPANTIFOL composition according to any of items

[50] to

[56] , wherein each pegylated liposome contains 1 to 1000 or 30 to 200 targeting moieties;

[58] An Lp-γPANTIFOL composition according to any of items

[39] to

[57] , further comprising one or more of an immunostimulant, a detectable marker, and maleimide, wherein the immunostimulant, the detectable marker, or the maleimide is bound to the PEG or outer surface of a liposome;

[59] An Lp-γPANTIFOL composition according to item

[58] , wherein the immunostimulant is at least one selected from the group consisting of protein immunostimulants, nucleic acid immunostimulants, chemoimmunostimulants, haptens, and adjuvants;

[60] Lp-γPANTIFOL composition as described in item

[58] or

[59] , wherein the immunostimulant is fluorescein, fluorescein isothiocyanate (FITC), DNP, beta-glucan, beta-1,3-glucan, beta-1,6-glucan, resolvin (e.g., D n-6DPA Or D n-3DPAAn Lp-γPANTIFOL composition comprising at least one selected from the group consisting of resolvin D, resolvin E, or T-series resolvins, oxidized low-density lipoproteins (e.g., OXPAC, PGPC), and Toll-like receptor (TLR) modulators such as erythritol lipids (e.g., E5564);

[61] An Lp-γPANTIFOL composition described in any of items

[58] to

[60] , wherein the immunostimulant and the detectable marker are the same;

[62] An Lp-γPANTIFOL composition according to any of items

[58] to

[61] , further comprising a hapten;

[63] Lp-γPANTIFOL composition as described in item

[62] , wherein the hapten comprises one or more of fluorescein or beta-1,6-glucan:

[64] An Lp-γPANTIFOL composition according to any of items

[12] to

[63] , further comprising at least one cryoprotective substance selected from the group consisting of mannitol, trehalose, sorbitol, and sucrose;

[65] Targeted compositions comprising any of the compositions described in items [1] to

[64] ;

[66] Non-targeting compositions comprising any of the compositions described in items [1] to

[49] ;

[67] An Lp-γPANTIFOL composition according to any of items

[12] to

[66] , further comprising carboplatin and / or pembrolizumab;

[68] A pharmaceutical composition comprising a liposomal gamma polyglutamine oxidized folate antimetabolitic agent composition described in any of items

[12] to

[67] ;

[69] A pharmaceutical composition comprising a gamma polyglutamine oxoxide antimetabolitic agent composition described in any of items [1] to [7];

[70] A composition described in any of items [1] to

[69] for use in the treatment of a disease;

[71] Use of any of the compositions described in items [1] to

[70] in the manufacture of a drug for the treatment of a disease;

[72] A method for treating or preventing a disease of which such treatment or prevention is required, comprising the step of administering a composition of any of items [1] to

[70] to the subject;

[73] A method for treating or preventing a disease in a subject requiring such treatment or prevention, comprising administering to a subject the liposomal gamma polyglutamine oxidized folate antimetabolitic composition described in any of

[12] to

[69] ;

[74] A method for killing overgrown cells, comprising the step of bringing the overgrown cells into contact with a composition described in any of items [1] to

[69] ;

[75] A method for killing overgrown cells, comprising the step of contacting the overgrown cells with a liposomal gamma polyglutamine oxidized folate antimetabolitic composition described in any of items

[12] to

[69] .

[76] Methods relating to item

[74] or

[75] , wherein the overgrowth cells are cancer cells, mammalian cells, and / or human cells;

[77] A method for treating cancer, comprising the step of administering an effective amount of any composition described in item [1] to

[69] to a subject who has or is at risk of having cancer;

[78] A method for treating cancer, comprising the step of administering an effective amount of a liposomal gamma polyglutamine oxidized folate antimetabolitic composition described in any of items

[12] to

[68] to a subject who has or is at risk of having cancer;

[79] Methods according to item

[77] or

[78] , wherein the cancer is a non-hematological malignancy, including, for example, lung cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, head and neck cancer, stomach cancer, gastrointestinal cancer, colorectal cancer, esophageal cancer, cervical cancer, liver cancer, kidney cancer, bile duct cancer, gallbladder cancer, bladder cancer, sarcoma (e.g., osteosarcoma), brain cancer, central nervous system cancer, and melanoma; and a method selected from, for example, hematological malignancies, including leukemia, lymphoma and other B-cell malignancies, myeloma and other plasma cell proliferation disorders;

[80] A method according to item

[77] or

[78] , wherein the cancer is selected from lung cancer, breast cancer, colorectal cancer, pancreatic cancer, gastric cancer, bladder cancer, head and neck cancer, ovarian cancer, and cervical cancer;

[81] A method according to item

[77] or

[78] , wherein the cancer is selected from colorectal cancer, lung cancer, breast cancer, head and neck cancer, and pancreatic cancer;

[82] A method according to item

[77] or

[78] , wherein the cancer is selected from colorectal cancer, breast cancer, ovarian cancer, lung cancer, head and neck cancer, pancreatic cancer, gastric cancer, and mesothelioma;

[83] A method for treating cancer, comprising the step of administering an effective amount of the Lp-γPANTIFOL composition described in any of items

[50] to

[66] to a subject having or at risk of having cancer cells expressing folate receptors bound by the targeting moiety on their surface;

[84] Maintenance therapy for subjects who are or have been receiving cancer therapy, comprising the step of administering an effective amount of any of the compositions described in items [1] to

[69] to a subject who is or has been receiving cancer therapy;

[85] Maintenance therapy for subjects currently receiving or formerly receiving cancer therapy, comprising the step of administering an effective amount of a liposomal gamma polyglutamine oxidative antimetabolitic composition described in any of items

[12] to

[69] to a subject currently receiving or formerly receiving cancer therapy;

[86] A method for treating an immune system disorder comprising the step of administering an effective amount of any composition described in item [1] to

[69] to a subject having or at risk of having an immune system disorder, wherein the immune system disorder is selected from inflammation (e.g., acute and chronic), systemic inflammation, rheumatoid arthritis, inflammatory bowel disease (IBD), Crohn's disease, dermatomyositis / polymyositis, systemic lupus erythematosus, and Takayasu's arteritis, and psoriasis;

[87] A method for treating an immune system disorder comprising the step of administering an effective amount of a liposomal gamma polyglutamine oxidized folate antimetabolitic composition described in any of items [8] to

[69] to a subject having or at risk of having an immune system disorder, wherein the immune system disorder is selected from inflammation (e.g., acute and chronic), systemic inflammation, rheumatoid arthritis, inflammatory bowel disease (IBD), Crohn's disease, dermatomyositis / polymyositis, systemic lupus erythematosus, and Takayasu's arteritis, and psoriasis;

[88] The following treatment methods: (a) A method for treating an infectious disease, comprising the step of administering an effective amount of any composition described in item [1] to

[69] to a subject having or at risk of having an infectious disease; (b) A method for treating an infectious disease, cardiovascular disease, metabolic disease, or another disease, comprising the step of administering an effective amount of any composition described in item [1] to

[69] to a subject having or at risk of having an infectious disease, cardiovascular disease, or another disease, wherein the disease is a member selected from atherosclerosis, cardiovascular disease (CVD), coronary artery disease, myocardial infarction, stroke, metabolic syndrome, gestational trophoblastic disease, and ectopic pregnancy; (c) A method for treating an autoimmune disease, comprising the step of administering an effective amount of any composition described in item [1] to

[69] to a subject having or at risk of having an autoimmune disease; (d) A method for treating rheumatoid arthritis, comprising the step of administering an effective amount of any composition described in item [1] to

[69] to a subject having or at risk of having rheumatoid arthritis; (e) A method for treating an inflammatory condition comprising the step of administering an effective amount of any composition described in item [1] to

[69] to a subject having or at risk of having inflammation, wherein the inflammation is acute, chronic, and / or systemic inflammation; or (f) A method for treating a skin disease, comprising the step of administering an effective amount of any composition described in item [1] to

[69] to a subject who has or is at risk of having a skin disease;

[89] A method for treating an infectious disease, comprising the step of administering an effective amount of a liposomal gamma polyglutamine oxidized folate antimetabolitic composition described in any of items

[12] to

[69] to a subject who has or is at risk of having an infectious disease;

[90] A method for delivering a gamma-polyglutamine oxidase antagonist to a tumor expressing folate receptors on its surface, comprising the step of administering an Lp-γPANTIFOL composition described in any of items [1] to

[69] to a subject having a tumor in an amount that delivers a therapeutically effective dose of the gamma-polyglutamine oxidase antagonist to the tumor;

[91] A method for preparing a gamma polyglutamate oxidase antagonist composition comprising a liposome gamma polyglutamate oxidase antagonist composition described in any of items

[12] to

[69] , comprising the steps of: forming a mixture comprising a liposome component and a gamma polyglutamate oxidase antagonist in solution; homogenizing the mixture in solution to form liposomes; and processing the mixture to form liposomes comprising a gamma polyglutamate oxidase antagonist.

[92] A method for preparing a gamma polyglutamate oxidase antagonist comprising a liposome gamma polyglutamate oxidase antagonist composition described in any of items

[12] to

[69] , comprising the steps of: forming a mixture in solution comprising a liposome component and a gamma polyglutamate oxidase antagonist; and processing the mixture to form liposomes comprising the gamma polyglutamate oxidase antagonist.

[93] A method according to item

[92] , wherein the step of processing the mixture comprises the step of homogenizing the mixture in solution to form liposomes.

[94] A method for preparing any of the compositions described in items

[50] to

[69] , comprising the steps of: forming a mixture in solution containing a liposome component and a gamma-polyglutamate oxidase antagonist; homogenizing the mixture in solution to form liposomes; processing the mixture to form liposomes that encapsulate and / or contain the gamma-polyglutamate oxidase antagonist; and imparting a targeting moiety on the surface of the liposomes, wherein the targeting moiety has specific affinity for at least one of folate receptor alpha (FR-α), folate receptor beta (FR-β), and folate receptor delta (FR-δ).

[95] A method for preparing any of the compositions described in items

[50] to

[69] , comprising the steps of: forming a mixture in solution containing a liposome component and a gamma-polyglutamate oxidase antagonist; processing the mixture to form liposomes that encapsulate and / or contain the gamma-polyglutamate oxidase antagonist; and imparting a targeting moiety on the surface of the liposomes, wherein the targeting moiety has specific affinity for at least one of folate receptor alpha (FR-α), folate receptor beta (FR-β), and folate receptor delta (FR-δ).

[96] A method relating to item

[95] , wherein the processing step comprises homogenizing a mixture in solution to form liposomes.

[97] Methods according to item

[92] , wherein the processing step comprises one or more steps of thin film hydration, extrusion, in-line mixing, ethanol injection technique, freeze-thaw method, reverse-phase evaporation method, dynamic high-pressure microfluidization, microfluidic mixing, double emulsion, freeze-dried double emulsion, 3D printing, membrane contactor method, and stirring; and / or

[98] A method according to any of items

[95] to

[97] , wherein the processing step comprises one or more steps of changing the size of liposomes by one or more steps of extrusion, high-pressure microfluidization, and / or sonication; and / or

[99] A method according to any of items

[91] to

[98] , wherein a starting material of at least 1% gamma polyglutamine oxidized folate antimetabolites is encapsulated or enclosed in liposomes.

[0089] I. Gamma polyglutamine oxidase antagonist (γPANTIFOL) Generally, this disclosure relates to gamma-polyglutamate oxidized folate antimetabolites (γPANTIFOL) compositions. Each γPANTIFOL composition contains at least one glutamyl group having a gamma-carboxyl group bond. These compositions are structurally different from L-gamma-polyglutamate oxidized folate antimetabolites (Lγ1PANTIFOL), which are produced in cells by the enzyme folyl-polygamma-glutamate synthase (FPGS) during folate antimetabolite therapy.

[0090] In some embodiments, the γPANTIFOL composition contains 2 to 20, 2 to 15, 2 to 10, 2 to 5, or more than 5 glutamyl groups (including the glutamyl groups of folate antagonists). In some embodiments, each glutamyl group in γPANTIFOL other than the glutamyl groups of folate antagonists has a gamma bond. In some embodiments, two or more glutamyl groups in γPANTIFOL have gamma bonds. In some embodiments, each glutamyl group in γPANTIFOL is L-type. In some embodiments, each glutamyl group in γPANTIFOL other than the glutamyl groups of folate antagonists is D-type. In some embodiments, γPANTIFOL contains two or more L-type glutamyl groups and one or more D-type glutamyl groups.

[0091] In some embodiments, the folate antagonist is selected from PMX, MTX, RTX, and LTX, or their stereoisomers.

[0092] In some embodiments, the folate antagonist is selected from the following: LV (etoposide), L-leucovorin (L-5-formyltetrahydrofolate); 5-CH3-THF, 5-methyltetrahydrofolate; FA, folate; PteGlu, pteroylglutamate (FA); MTX, methotrexate; 2-dMTX, 2-desamino-MTX; 2-CH3-MTX, 2-desamino-2-methyl-MTX; AMT, aminopterin; 2-dAMT, 2-desamino-AMT; 2-CH3-AMT, 2-desamino-2-methyl-AMT; 10-EdAM, 10-ethyl-10-deazaaminopterin; PT523, N alpha-(4-amino-4-deoxypteroyl)-N Delta-(hemiphthaloyl)-L-ornithine;DDATHF (lometrexol), 5,10-dideaza-5,6,7,8-tetrahydrofolate;5-d(i)H4PteGlu, 5-deaza-5,6,7,8-tetrahydroisofolate;N9-CH3-5-d(i)H4PteGlu, N9-methyl-5-deaza-5,6,7,8-tetrahydroisofolate;5-dPteHCysA,N Alpha-(5-deazapteroyl)-L-homocysteic acid;5-dPteAPBA,N Alpha-(5-deazapteroyl)-DL-2-amino-4-phosphonobutyric acid;5-dPteOrn,N Alpha-(5-deazapteroyl)-L-ornithine;5-dH4PteHCysA,N Alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-L-homocysteic acid; 5-dH4PteAPBA,N Alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-DL-2-amino-4-phosphobutanoic acid; 5-dH4PteOro,N Alpha-(5-dideaza-5,6,7,8-tetrahydropteroyl)-L-ornithine;CB3717, N10-propargyl-5,8-dideazafolate;ICI-198,583, 2-desamino-2-methyl-N10-propargyl-5,8-dideazafolate;4-H-ICI-198,583, 4-deoxy-ICI-198,583:4-OCH3-ICI-198,583, 4-methoxy-ICI-198,583 Glu-to-Val-ICI-198,583;Valine-ICI-198;583;Glu-to-Sub-ICI-198,583,2-aminosuberic acid-ICI-198,583;7-CH3-ICI-198,583,7-methyl-ICI-198,583;ZD1694,N-[5(N-(3,4-dihydro-2-methyl-4-oxoquinazoline-6-yl-methyl)amino)2-thienyl)]-L-glutamic acid;2-NH2- ZD1694, 2-amino-ZD1694; BW1843U89, (S)-2[5-(((1,2-dihydro-3-methyl-1-oxobenzo(f)quinazolin-9-yl)methyl)amino-)-1-oxo-2-isoindolinyl]-glutaric acid; LY231514, N-(4-(2-(2-amino-4,7-dihydro-4-oxo-3H-pyrrolo[2,3 -D]pyrimidine-5-yl)ethyl)benzoyl]-L-glutamic acid; IAHQ, 5,8-dideazaisofolate; 2-dIAHQ, 2-desamino-IAHQ; 2-CH3-dIAHQ, 2-desamino-2-methyl-IAHQ; 5-d(i)PteGlu, 5-deazaisofolate; N9-CH3-5-d(i)PteGlu, N9-methyl-5-deazaisofolate; N9-CHO-5-d(i)PteGlu, N9-formyl-5-deazaisofolate; AG337, 3,4-dihydro-2-amino-6-methyl-4-oxo-5-(4-pyridylthio)quanazoline; and AG377, 2,4-diamino-6[N-(4-(phenisulfonyl)benzyl)ethyl)amino]quinazoline; or their stereoisomers. ;

[0093] In some embodiments, the folate antagonist is selected from methotrexate, larcitrexed, previtrexed, pemetrexed, lometrexol (LTX; 5,10-dideazatetrahydrofolate), cyclopenta[g]quinazoline with a dipeptide ligand, CB3717, CB300945, or their stereoisomers such as 6-R, S-BGC945 (ONX-0801), CB300638, and BW1843U89.

[0094] In some embodiments, the folate antagonist is a 6-substituted pyrrolo[2,3-d]pyrimidine benzoyl antiphorate. In some embodiments, the folate antagonist is a 6-substituted pyrrolo[2,3-d]pyrimidine benzoyl antiphorate having carbon bridges of 1 to 6 carbon lengths (e.g., a compound having the structure of (I), n1=1 to 6). In some embodiments, the folate antagonist is a 6-substituted thieno[2,3-d]pyrimidine benzoyl antiphorate having carbon bridges of 2 to 8 carbon lengths (e.g., a compound having the structure of formula (II), n2=7 to 13). In some embodiments, the folate antagonist is a 6-substituted pyrrolo[2,3-d]pyrimidine antiphorate having carbon bridges of 2 to 8 carbon lengths, with the benzoyl moiety substituted with thienoyl (e.g., a compound having the structure of formula (III), n1=1 to 6). In some embodiments, the folate antagonist has a structure according to one of formulas (I) to (III), where x = 4, 5, 2 to 10, 4 to 6, or greater than 5. [ka]

[0095] In some embodiments, the folate antagonist is selected from the following: methotrexate derivatives containing an indoline ring and modified ornithine, methotrexate derivatives containing an indoline ring and modified glutamic acid, methotrexate derivatives containing an alkyl-substituted benzene ring C, methotrexate derivatives partially containing benzoxazine, methotrexate derivatives partially containing benzothiazine, 10-deazaaminopterin analogs, 5-deazaaminopterin methotrexate analogs, 5,10-dideazaaminopterin methotrexate analogs, and Dorin-containing methotrexate derivatives, lipophilic amide methotrexate derivatives, L-threo-(2S,4S)-4-fluoroglutamic acid-containing methotrexate analogs, DL-3,3-difluoroglutamic acid-containing methotrexate analogs, methotrexate tetrahydroquinazoline analogs, N-(ac-aminoacyl)methotrexate derivatives, biotin methotrexate derivatives, D-glutamic acid methotrexate analogs, D-erythro,threo-4-fluoroglutamic acid methotrexate analogs, β,γ-methano Methotrexate analogs, 10-deazaaminopterin (10-EDAM) analogs, γ-tetrazolemethotrexate analogs, N-(L-α-aminoacyl)methotrexate derivatives, meta-isomers of aminopterin, ortho-isomers of aminopterin, hydroxymethylmethotrexate, γ-fluoromethotrexate, polyglutamylmethotrexate derivatives, gem-diphosphonate-methotrexate analogs (see, for example, International Application No. 1988 / 06158, the contents of which are incorporated herein by reference in their entirety), α-substituted methotrexate analogs, γ-substituted methotrexate analogs, 5-methyl-5-deazamethotrexate analogs (see, for example, U.S. Patent No. 4,725,See issue 687, which is incorporated herein by reference in its entirety), N δ-acyl-N α-(4-amino-4-deoxypteroyl)-L-ornithine derivatives, 8-deazamethotrexate analogs, asibicinmethotrexate analogs, polymerplatinolmethotrexate derivatives, methotrexate-γ-dimyristoylphophatidylethanolamine, methotrexate polyglutamate analogs, poly-γ-glutamylmethotrexate derivatives, deoxyuridilatemethotrexate derivatives, iodoacetyllysinemethot Lexate analogs, 2,ω-diaminoalkanoid acid-containing methotrexate analogs, polyglutamate derivatives, 5-methyl-5-deaza analogs, quinazoline methotrexate analogs, pyrazine methotrexate analogs, cysteine ​​or homocysteine ​​methotrexate analogs (see, for example, U.S. Patent No. 4,490,529 and EPA Patent No. 0142220, the contents of which are incorporated herein by reference in their entirety), γ-tert- Butylmethotrexate ester, fluoride methotrexate analog, folic acid methotrexate analog, phosphonoglutamic acid analog, poly(L-lysine)methotrexate complex, dilysine or trilysine methotrexate derivatives, 7-hydroxymethotrexate, poly-γ-glutamylmethotrexate analog, 3',5'-dichloromethotrexate, diazoketone or chloromethyl ketone methotrexate analog, 10-propargyl Luaminopterin, alkylmethotrexate homologs, lectin derivatives of methotrexate, polyglutamate methotrexate derivatives, halogenated methotrexate derivatives, 8-alkyl-7,8-dihydro analogs, 7-methylmethotrexate derivatives, dichloromethotrexate, lipophilic methotrexate derivatives, 3',5'-dichloromethotrexate, deazamethopterin analogs, and MX068; or their stereoisomers.

[0096] In some embodiments, the folate antagonist is given by formula (IV): [ka] The formula has the following properties: X = CH2, C2H4, or O(CH2)3O; R1 = Me or Et; R2 = H, Cl, F, OH, or R2 = R3; and R3 = H, Cl, F, OH, Me, or Br.

[0097] In some embodiments, the folate antagonist has formula (IV), where X=CH2;R1=Me or Et;R2=H, Cl, F, OH, or R2=R3; and R3=H, Cl, F, OH, Me, or Br. In some embodiments, X=CH2;R1=Me;R2=H, Cl, F, OH; and R3=H, Cl, Me, or Br. In some embodiments, X=O(CH2)3O;R1=Me; and R2=R3=H.

[0098] In some embodiments, the folate antagonist is given by formula (V): [ka] It has X = C2H4, C4H8, C6H 12 O(CH2)2O or O(CH2)3O; R1=H or Cl, or R2=R3; and R3=H or Cl.

[0099] In some embodiments, the folate antagonist has formula (V), where X = C2H4 and R1 = R2 = H or Cl. In some embodiments, X = C2H4; R1 = Cl; and R2 = H. In some embodiments, X = C4H8; and R1 = R2 = H. In some embodiments, X = C6H 12 ; and R1=R2=H.

[0100] In some embodiments, the folate antagonist is given by formula (VI): [ka] It has X=CH2 or C2H4; Y=2,5-thiophene; and R=CH2F, Cn, Et, Me, or CH2OH.

[0101] In some embodiments, the folate antagonist has formula (VI), where X = CH2; Y = 2,5-thiophene; and R = H2F, Cn, Et, or CH2OH. In some embodiments, X = C2H4; Y = 2,5-thiophene; and R = Me.

[0102] In some embodiments, the folate antagonist is given by formula (VII): [ka] The formula has the following properties: X = N or CH, Y = NH2, CH3, or H; and R = CH3, CHO, or H.

[0103] In some embodiments, the folate antagonist has formula (VII), where (a) X=N; Y=NH2; and R=H; (b) X=N; H=NH2; and R=CH3; (c) X=N; Y=NH2; and R=CHO; (d) X=CH, Y=NH2, R=H; (e) X=CH, Y=H, R=H; or (f) X=CH, Y=CH3, and R=H.

[0104] In some embodiments, the folate antagonist is given by formula (VIII): [ka] It has the following properties: A=NH, NCH3, or CH2.

[0105] In some embodiments, the folate antagonist is given by formula (IX): [ka] The following conditions are met: (a) X=OH; R=H; and Y=Glu, (b) X=OCH3; R=H; and Y=Glu, (c) X=OH; R=H; and Y=valine, (d) X=OH; R=H; and Y=sverate, or (e) X=OH; R=CH3; and Y=Glu.

[0106] In further embodiments, the folate antagonist is a cyclopenta[g]quinazoline derivative. In some embodiments, the cyclopenta[g]quinazoline derivative is N-{N-{4-[N-(2-methyl-4-oxo-3,4,7,8-tetrahydro-6H-cyclopenta[g]quinazoline-6-yl)-N-(propa-2-inyl)amino]benzoyl}-L-γ-glutamyl}-D-glutamic acid; or N-{N-{4-[N-(2-hydroxymethyl-4-oxo-3,4,7,8-tetrahydro-6H-cyclopenta[g]quinazoline-6-yl)-N-(propa-2-inyl)amino]benzoyl}-L-γ-glutamyl}-D-glutamic acid; or a pharmaceutically acceptable salt or ester thereof.

[0107] In some embodiments, the folate antagonist is given by formula (X): [ka] It has such that R1 is H, amino, C1-4 alkyl, C1-4 alkoxy, C1-4 hydroxyalkyl, or C1-4 fluoroalkyl; R2 is hydrogen, C1-4 alkyl, C3-4 alkenyl, C3-4 alkynyl, C2-4 hydroxyalkyl, C2-4 halogenoalkyl, or C1-4 cyanoalkyl; Ar is phenylene, thiophenediyl, thiazolediyl, pyridinediyl or pyrimidinediyl, which optionally have one or two substituents selected from halogeno, hydroxy, amino, nitro, cyano, trifluoromethyl, C1-4 alkyl and C1-4 alkoxy, and R3 is one of the groups in the following formula: -NHCH(CO2H)-A1-Y1-NH-A3-Y3, or R3 is an alpha or gamma carboxyl-linked L- or D-glutamyl group.

[0108] In some embodiments, the folate antagonist has formula (X), where R1 is a C1-4 alkyl or C1-4 hydroxyalkyl (e.g., methyl or hydroxymethyl); R2 is (a) methyl, ethyl, propyl, propa-2-enyl, propa-2-inyl, 2-hydroxyethyl, 2-fluoroethyl, 2-bromoethyl or 2-cyanoethyl, (b) methyl or (c) propa-2-inyl; Ar is 1,4-phenylene (e.g., 2-fluoro substituent, e.g., 2-fluoro-1,4-phenylene or 2,6-difluoro-1,4-phenylene), thiophene-2,5-diyl, thiazole-2,5-diyl or pyridine-2,5-diyl, having one or two substituents selected from chloro and fluoro.

[0109] In some embodiments, the folate antagonist has formula (X), where R1 is methyl or hydroxymethyl; R2 is methyl or propan-2-inyl; and Ar is 1,4-phenylene or 2,6-difluoro-1,4-phenylene or 1,4-phenylene having a 2-fluoro substituent such as 2-fluoro-1,4-phenylene, or pyridine 2,5-diyl. In some embodiments, Ar is 1,4-phenylene or 2-fluoro-1,4-phenylene.

[0110] In other embodiments, the gamma-polyglutamine oxidase antagonist is cyclopenta[g]quinazoline, as disclosed in International Publication No. 2009 / 115776, International Publication No. 2003 / 020300, International Publication No. 2003 / 020706, International Publication No. 2003 / 020748, Gibbs et al., Cancer Research 65(15):11721-11728 (2005), and Bavetsias et al., Tetrahedron 63(7):1537-1543 (2007). The contents of each of these are incorporated herein by reference in their entirety.

[0111] In some embodiments, the gamma-polyglutamate folate antagonist is diglutamine-oxidized. That is, the gamma-polyglutamate folate antagonist contains one additional glutamyl group (γANTIFOL-PG1) in addition to the glutamyl group in the folate antagonist, and this additional glutamyl group is bonded to the glutamyl group in the folate antagonist via a gamma bond. In some embodiments, each glutamyl group in the gamma-diglutamate folate antagonist is L-type. In other embodiments, the gamma-diglutamate folate antagonist contains D-type glutamyl groups.

[0112] In some embodiments, the gamma-polyglutamate folate antagonist is triglutamate oxidized. That is, the gamma-polyglutamate folate antagonist contains two gamma-glutamyl groups in addition to the glutamyl group in the folate antagonist (γANTIFOL-PG2). In some embodiments, each of the two additional glutamyl groups has a gamma bond. In other embodiments, one of the two glutamyl groups has a gamma bond and the other glutamyl group has a gamma bond. In some embodiments, each glutamyl group in the gamma-triglutamate folate antagonist is L-type. In other embodiments, the gamma-triglutamate folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group in the gamma-triglutamate folate antagonist, other than the gamma-glutamyl group in the folate antagonist, is D-type. In further embodiments, the γ-triglutamine oxidase folate antagonist comprises a D-type glutamyl group and two or more L-type glutamyl groups.

[0113] In some embodiments, the gamma-polyglutamine-oxidized folate antagonist is tetraglutamine-oxidized and therefore contains three γ-glutamyl groups in addition to the glutamyl group of the folate antagonist (γANTIFOL-PG3). In some embodiments, the gamma-tetraglutamine-oxidized folate antagonist contains two or more L-type γ-glutamyl groups. In further embodiments, each γ-glutamyl group of the gamma-tetraglutamine-oxidized folate antagonist is L-type. In other embodiments, the gamma-tetraglutamine-oxidized folate antagonist contains D-type γ-glutamyl groups. In some embodiments, the gamma-tetraglutamine-oxidized folate antagonist contains two D-type γ-glutamyl groups. In some embodiments, each glutamyl group of the gamma-tetraglutamine-oxidized folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the tetraglutamine oxidase folate antimetabolite comprises a D-type γ-glutamyl group and two or more L-type γ-glutamyl groups.

[0114] In some embodiments, the gamma-polyglutamine-oxidized folate antagonist contains a chain of four γ-glutamyl groups that are pentaglobin-oxidized (γANTIFOL-PG4) and bonded to a glutamyl group in the folate antagonist. In some embodiments, the gamma-pentaglutamine-oxidized folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the gamma-pentaglutamine-oxidized folate antagonist is L-type. In other embodiments, the gamma-pentaglutamine-oxidized folate antagonist contains a D-type glutamyl group. In some embodiments, the gamma-tetraglutamine-oxidized folate antagonist contains two or three D-type γ-glutamyl groups. In further embodiments, each γ-glutamyl group in the gamma-pentaglutamine-oxidized folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the pentaglutamine oxidase antagonist comprises a D-type γ-glutamyl group and two or more L-type γ-glutamyl groups.

[0115] In some embodiments, the gamma-polyglutamine-oxidized folate antagonist contains a chain of five γ-glutamyl groups bonded to a glutamyl group in the folate antagonist, after being hexaglutamine-oxidized (γANTIFOL-PG5). In some embodiments, the gamma-hexaglutamine-oxidized folate antagonist contains two or more L-type γ-glutamyl groups. In further embodiments, each glutamyl group in the gamma-hexaglutamine-oxidized folate antagonist is L-type. In other embodiments, the gamma-hexaglutamine-oxidized folate antagonist contains D-type γ-glutamyl groups. In some embodiments, the gamma-tetraglutamine-oxidized folate antagonist contains two, three, four, or five D-type γ-glutamyl groups. In further embodiments, each glutamyl group in the gamma-hexaglutamine-oxidized folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the hexaglutamine oxidase folate antagonist comprises a D-type γ-glutamyl group and two or more L-type γ-glutamyl groups.

[0116] In some embodiments, the gamma-polyglutamine-oxidized folate antagonist is heptaglutamine-oxidized (γANTIFOL-PG6) and therefore contains a chain of six γ-glutamyl groups bonded to a glutamyl group in the folate antagonist. In some embodiments, the gamma-heptaglutamine-oxidized folate antagonist contains two or more L-type γ-glutamyl groups. In further embodiments, each γ-glutamyl group in the gamma-heptaglutamine-oxidized folate antagonist is L-type. In other embodiments, the gamma-heptaglutamine-oxidized folate antagonist contains D-type γ-glutamyl groups. In some embodiments, the gamma-tetraglutamine-oxidized folate antagonist contains two, three, four, five, or six D-type γ-glutamyl groups. In further embodiments, each γ-glutamyl group in the gamma-heptaglutamine-oxidized folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the heptaglutamine oxidase antagonist comprises a D-type γ-glutamyl group and two or more L-type γ-glutamyl groups.

[0117] In some embodiments, the gamma-polyglutamine-oxidized folate antagonist is octaglutamine-oxidized (γANTIFOL-PG7) and therefore contains a chain of seven γ-glutamyl groups bonded to a glutamyl group in the folate antagonist. In some embodiments, the gamma-octaglutamine-oxidized folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the gamma-octaglutamine-oxidized folate antagonist is L-type. In other embodiments, the gamma-octaglutamine-oxidized folate antagonist contains D-type glutamyl groups. In some embodiments, the gamma-octaglutamine-oxidized folate antagonist contains two, three, four, five, six, or seven D-type γ-glutamyl groups. In further embodiments, each glutamyl group in the gamma-octaglutamine-oxidized folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the octaglutamine oxoxide antimetabolite comprises a D-type glutamyl group and two or more L-type glutamyl groups.

[0118] In some embodiments, the gamma-polyglutamine-oxidized folate antagonist contains a chain of eight γ-glutamyl groups bonded to a glutamyl group in the folate antagonist after being nonaglutamine-oxidized (γANTIFOL-PG8). In some embodiments, the gamma-nonaglutamine-oxidized folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the gamma-nonaglutamine-oxidized folate antagonist is L-type. In other embodiments, the gamma-nonaglutamine-oxidized folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group in the gamma-nonaglutamine-oxidized folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the nonaglutamine-oxidized folate antagonist contains a D-type γ-glutamyl group and two or more L-type γ-glutamyl groups.

[0119] In some embodiments, the gamma-polyglutamate folate antagonist is decadaglutamine-oxidized (γANTIFOL-PG9) (i.e., it contains a chain of nine γ-glutamyl groups bonded to a glutamyl group in the folate antagonist). In some embodiments, the gamma-decaglutamate folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the gamma-decaglutamate folate antagonist is L-type. In other embodiments, the gamma-decaglutamate folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group in the gamma-decaglutamate folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the decadaglutamate folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups.

[0120] In some embodiments, the gamma-polyglutamate folate antagonist is undecaglutamine-oxidized (γANTIFOL-PG10) and contains a chain of 10 γ-glutamyl groups bonded to a glutamyl group in the folate antagonist. In some embodiments, the gamma-undecaglutamate folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the gamma-undecaglutamate folate antagonist is L-type. In other embodiments, the gamma-undecaglutamate folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group in the gamma-undecaglutamate folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the undecaglutamate folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups.

[0121] In some embodiments, the gamma-polyglutamine-oxidized folate antagonist is dodecaglutamine-oxidized (γANTIFOL-PG11) and contains a chain of 11 γ-glutamyl groups bonded to a glutamyl group in the folate antagonist. In some embodiments, the gamma-dodeca-glutamine-oxidized folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the gamma-dodeca-glutamine-oxidized folate antagonist is L-type. In other embodiments, the gamma-dodeca-glutamine-oxidized folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group in the gamma-dodeca-glutamine-oxidized folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the dodeca-glutamine-oxidized folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups.

[0122] In some embodiments, the gamma-polyglutamine-oxidized folate antagonist is tridecaglutamine-oxidized (γANTIFOL-PG12) and contains a chain of 12 γ-glutamyl groups bonded to a glutamyl group in the folate antagonist. In some embodiments, the gamma-trideca-glutamine-oxidized folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the gamma-trideca-glutamine-oxidized folate antagonist is L-type. In other embodiments, the gamma-trideca-glutamine-oxidized folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group in the gamma-trideca-glutamine-oxidized folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the tridecaglutamine-oxidized folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups.

[0123] In some embodiments, the gamma-polyglutamine-oxidized folate antagonist is tetradecaglutamine-oxidized (γANTIFOL-PG13) and contains a chain of 13 γ-glutamyl groups bonded to a glutamyl group in the folate antagonist. In some embodiments, the gamma-tetradecaglutamine-oxidized folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the gamma-tetradecaglutamine-oxidized folate antagonist is L-type. In other embodiments, the gamma-tetradecaglutamine-oxidized folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group in the gamma-tetradecaglutamine-oxidized folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the tetradecaglutamine-oxidized folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups.

[0124] In some embodiments, the gamma-polyglutamine-oxidized folate antagonist is pentadeca-glutamine-oxidized (γANTIFOL-PG14) and contains a chain of 14 γ-glutamyl groups bonded to a glutamyl group in the folate antagonist. In some embodiments, the gamma-pentadeca-glutamine-oxidized folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the gamma-pentadeca-glutamine-oxidized folate antagonist is L-type. In other embodiments, the gamma-pentadeca-glutamine-oxidized folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group in the gamma-pentadeca-glutamine-oxidized folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the pentadeca-glutamine-oxidized folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups.

[0125] In some embodiments, the gamma-polyglutamine-oxidized folate antagonist is hexadeca-glutamine-oxidized (γANTIFOL-PG15) and contains a chain of 15 γ-glutamyl groups bonded to a glutamyl group in the folate antagonist. In some embodiments, the gamma-hexadeca-glutamine-oxidized folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the gamma-hexadeca-glutamine-oxidized folate antagonist is L-type. In other embodiments, the gamma-hexadeca-glutamine-oxidized folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group in the gamma-hexadeca-glutamine-oxidized folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the hexadeca-glutamine-oxidized folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups.

[0126] In other embodiments, the gamma-polyglutamine-oxidized folate antagonist is heptadeca-glutamine-oxidized (γANTIFOL-PG16) and contains a chain of 16 γ-glutamyl groups bonded to a glutamyl group in the folate antagonist. In some embodiments, the gamma-heptadeca-glutamine-oxidized folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the gamma-heptadeca-glutamine-oxidized folate antagonist is L-type. In other embodiments, the gamma-heptadeca-glutamine-oxidized folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group in the gamma-heptadeca-glutamine-oxidized folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the heptadeca-glutamine-oxidized folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups.

[0127] In some embodiments, the gamma-polyglutamate folate antagonist is octadeca-glutamate oxidized (γANTIFOL-PG17) and contains a chain of 17 γ-glutamyl groups bonded to a glutamyl group in the folate antagonist. In some embodiments, the gamma-octadeca-glutamate folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the gamma-octadeca-glutamate folate antagonist is L-type. In other embodiments, the gamma-octadeca-glutamate folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group in the gamma-octadeca-glutamate folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the octadeca-glutamate folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups.

[0128] In some embodiments, the gamma-polyglutamine-oxidized folate antagonist is nonadeca-glutamine-oxidized (γANTIFOL-PG18) and contains a chain of 18 γ-glutamyl groups bonded to a glutamyl group in the folate antagonist. In some embodiments, the gamma-nonadeca-glutamine-oxidized folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the gamma-nonadeca-glutamine-oxidized folate antagonist is L-type. In other embodiments, the gamma-nonadeca-glutamine-oxidized folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group in the gamma-nonadeca-glutamine-oxidized folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the nonadeca-glutamine-oxidized folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups.

[0129] In some embodiments, the gamma-polyglutamate folate antagonist is eicosa-glutamate oxidized (γANTIFOL-PG19) and contains a chain of 19 γ-glutamyl groups bonded to a glutamyl group in the folate antagonist. In some embodiments, the gamma-mycosaglutamate folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the gamma-mycosaglutamate folate antagonist is L-type. In other embodiments, the gamma-mycosaglutamate folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group in the gamma-mycosaglutamate folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the eicosa-glutamate folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups.

[0130] In some embodiments, the gamma-polyglutamate folate antagonist is henicosaglutamate-oxidized (γANTIFOL-PG20) and contains a chain of 20 γ-glutamyl groups bonded to a glutamyl group in the folate antagonist. In some embodiments, the gamma-henicosaglutamate folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the gamma-henicosaglutamate folate antagonist is L-type. In other embodiments, the gamma-henicosaglutamate folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group in the gamma-henicosaglutamate folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the henicosaglutamate folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups.

[0131] In some embodiments, the gamma-polyglutamic folate antagonist contains 4 to 7 glutamyl groups bound to the folate antagonist (i.e., γANTIFOL-PGn, n=4 to 7), each of the 4 to 7 bound glutamyl groups having a gamma bond. In some embodiments, each of the 4 to 7 bound glutamyl groups is L-type. In other embodiments, each of the 4 to 7 bound glutamyl groups is D-type. In other embodiments, the 4 to 7 bound glutamyl groups are L-type and D-type.

[0132] In some embodiments, the gamma polyglutamic folate antimetabolite (γPANTIFOL) contains a total of 1 to 15, 1 to 10, 2 to 15, 2 to 10, 3 to 15, 3 to 10, 3 to 6, 3 to 5, 4 to 10, 4 to 7, or 4 to 6 glutamyl groups, or any range between these, including the glutamyl groups of the folate antimetabolite. In some embodiments, each glutamyl group in γPANTIFOL other than the glutamyl group in the folate antimetabolite has a gamma bond. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 glutamyl groups in γPANTIFOL have gamma bonds. In some embodiments, γPANTIFOL contains L-type and D-type γ-glutamyl groups. In some embodiments, each glutamyl group in the polyglutamate structure of the polyglutamic folate antimetabolite is L-type. In some embodiments, each glutamyl group in γPANTIFOL, other than the glutamyl group in the folate antagonist, is of the D type. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 glutamyl groups in γPANTIFOL are of the L type. In another embodiment, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 glutamyl groups in γPANTIFOL are of the D type.

[0133] In some embodiments, the gamma polyglutamate oxidase folate antagonist (γPANTIFOL) contains a total of 2 to 20, 2 to 15, 2 to 10, 2 to 5, or any range in between, including the glutamyl group of the folate antagonist. In some embodiments, each glutamyl group in γPANTIFOL is L-type. In some embodiments, each glutamyl group in γPANTIFOL other than the glutamyl group in the folate antagonist is D-type. In one embodiment, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 glutamyl groups in γPANTIFOL are L-type. In another embodiment, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 glutamyl groups in γPANTIFOL are of the D type.

[0134] In some embodiments, the gamma-polyglutamate folate antagonist contains a total of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 glutamyl groups in addition to the glutamyl group of the folate antagonist.

[0135] In some embodiments, a total of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 glutamyl groups in the gamma polyglutamate folate antagonist are L-type, D-type, or L-type and D-type. In some embodiments, each glutamyl group in the gamma polyglutamate folate antagonist is L-type. In other embodiments, each glutamyl group in the gamma polyglutamate folate antagonist, other than the glutamyl group of the folate antagonist, is D-type. In alternative embodiments, at least two glutamyl groups in the gamma polyglutamate folate antagonist are L-type, and at least one glutamyl group in the gamma polyglutamate folate antagonist is D-type. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 glutamyl groups in the gamma polyglutamate folate antimetabolite are L-type. In other embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 glutamyl groups in the gamma polyglutamate folate antimetabolite are D-type.

[0136] In further embodiments, the gamma-polyglutamate folate antagonist contains 20-100, 20-75, 20-50, 20-40, 20-30, 20-25, or more than 100 glutamyl groups, or any range between these. In some embodiments, each glutamyl group of the gamma-polyglutamate folate antagonist is L-type. In other embodiments, each glutamyl group of the gamma-polyglutamate folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In alternative embodiments, at least two glutamyl groups in the gamma-polyglutamate folate antagonist are L-type, and at least one glutamyl group in the gamma-polyglutamate folate antagonist is D-type.

[0137] In further embodiments, the provided composition comprises a gamma-polyglutamate folate antagonist comprising 1, 2, 3, 4, 5, 6, 7, 8, 9, 1-10, or 1-20 glutamyl groups having gamma bonds. In some embodiments, the gamma-polyglutamate folate antagonist comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 1-10, or 1-20 L-type glutamyl groups. In some embodiments, the gamma-polyglutamate folate antagonist comprises 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 1-10, or 1-20 D-type glutamyl groups. In some embodiments, the gamma polyglutamic oxidase folate antagonist comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 1-10, or 1-20 L-type glutamyl groups and 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 1-10, or 1-20 D-type glutamyl groups.

[0138] In some embodiments, the gamma-polyglutamate oxidase folate antagonist compositions provided herein may be modified to include one or more additional glutamyl groups, i.e., the compositions may function as substrates for FPGAs (folyl polyglutamate synthetase). Reagents, assays, and reagents for measuring the ability of gamma-polyglutamate oxidase folate antagonist compositions to act as substrates for FPGAs (e.g., human FPGAs or rat liver FPGAs) are readily available and can be performed as part of routine procedures.

[0139] In some embodiments, the rate of uptake by hepatocytes of the naked gamma-pantifol composition disclosed herein (e.g., gamma-pantifol not bound to a delivery carrier) is significantly reduced under physiological conditions compared to the uptake rate of folate antimetabolites. In some embodiments, the rate of hepatocyte uptake of the naked gamma-pantifol composition is 30%, 20%, 15%, or less than 10% compared to the rate of folate antimetabolites. In further embodiments, the rate of efflux (transport) of the gamma-pantifol composition disclosed herein from hepatocytes occurs at a significantly slower rate (30%, 20%, 15%, or less than 10%) compared to folate antimetabolites.

[0140] In some embodiments, the gamma-polyglutamine-oxidized folate antagonist compositions provided herein have higher cytotoxicity against hyperproliferating cells than folate antagonists. In some embodiments, the hyperproliferating cells are cancer cells. In some embodiments, the hyperproliferating cells are colorectal cancer cells, colon cancer cells, breast cancer cells, or ovarian cancer cells. In some embodiments, the cancer cells are mesothelioma cells or non-small cell lung cancer cells. In some embodiments, cytotoxicity is measured by an in vitro assay. In some embodiments, the gamma-polyglutamine-oxidized folate antagonist is a hexaglutamine-oxidized folate antagonist.

[0141] In some embodiments, the gamma-polyglutamate-oxidized folate antimetabolites provided herein have lower toxic side effects than folate antimetabolites. In some embodiments, the gamma-polyglutamate-oxidized folate antimetabolites provided herein are less toxic to non-overgrowth cells than folate antimetabolites. In some embodiments, the gamma-polyglutamate-oxidized folate antimetabolites provided herein are less toxic to neutrophils, hepatocytes, or colon epithelial cells than folate antimetabolites. In some embodiments, neutrophils are human neutrophils, differentiated human neutrophils, or neutrophils differentiated from CD34+ cells. In some embodiments, hepatocytes are AML12 hepatocytes. In some embodiments, colon epithelial cells are CCD841 colon epithelial cells. In some embodiments, toxicity is measured by an in vitro assay. In some embodiments, the gamma-polyglutamate-oxidized folate antimetabolite is a hexaglutamate-oxidized folate antimetabolite.

[0142] In some embodiments, the gamma-polyglutamate-oxidized folate antimetabolites provided herein have lower toxic side effects compared to folate antimetabolites. In some embodiments, the gamma-polyglutamate-oxidized folate antimetabolites provided herein result in fewer or less severe toxic side effects than folate antimetabolites in in vivo assays. In some embodiments, the in vivo assay is performed in an in vivo mouse model. In some embodiments, the gamma-polyglutamate-oxidized folate antimetabolites provided herein result in fewer or less severe hematological or hepatotoxic side effects than folate antimetabolites. In some embodiments, hematological side effects are assessed by mean neutrophils, mean leukocytes, or mean platelet counts. In some embodiments, hepatotoxic side effects are assessed by measuring serum aspartate aminotransferase (AST), serum alanine aminotransferase (ALT), and / or serum albumin levels. In some embodiments, the in vivo assay involves administering a gamma-polyglutamine oxidase antagonist composition at a dose of 40 mg / kg or 80 mg / kg once weekly for four weeks. In some embodiments, the gamma-polyglutamine oxidase antagonist is a hexaglutamine oxidase antagonist.

[0143] In some embodiments, treatment with the gamma-polyglutamate oxidase folate antagonist compositions provided herein does not induce significant hematological or hepatic toxic side effects in in vivo mouse models. In some embodiments, hematological side effects are assessed by mean neutrophil, mean leukocyte, or mean platelet count. In some embodiments, hepatic toxic side effects are assessed by measuring serum aspartate aminotransferase (AST), serum alanine aminotransferase (ALT), and / or serum albumin levels. In some embodiments, the gamma-polyglutamate oxidase folate antagonist compositions provided herein do not significantly decrease mean neutrophil, mean leukocyte, or mean platelet count. In some embodiments, the gamma-polyglutamate oxidase folate antagonist compositions provided herein do not significantly increase serum aspartate aminotransferase (AST) and serum alanine aminotransferase (ALT) levels. In some embodiments, the gamma-polyglutamate oxidase folate antagonist compositions provided herein do not significantly decrease serum albumin levels. In some embodiments, the in vivo assay involves administering a gamma-polyglutamine oxidase antagonist composition at a dose of 40 mg / kg or 80 mg / kg once weekly for four weeks. In some embodiments, the gamma-polyglutamine oxidase antagonist is a hexaglutamine oxidase antagonist.

[0144] In some embodiments, the gamma-polyglutamine oxidase folate antagonist composition does not contain a fluorine atom. In some embodiments, the gamma-polyglutamine oxidase folate antagonist composition does not contain a 4-fluoroglutamyl group.

[0145] The gamma polyglutamine oxidized folate antimetabolites (γPANTIFOL) compositions and their uses are further described in International Application PCT / US2017 / 046667 and U.S. Patent Applications 62 / 630,824, 62 / 630,613, 62 / 630,713, 62 / 630,620, 62 / 627,733, 62 / 630,625, 62 / 630,652, 62 / 627,732, 62 / 636,289, 62 / 630,751, 62 / 630,821, 62 / 627,741, and 62 / 583,432, respectively. Each of these disclosures is incorporated herein by reference in its entirety.

[0146] A. Gamma polyglutamine oxoxide antimetabolitic antagonist analogs and derivatives This disclosure also includes gamma-polyglutamine-oxidized folate antimetabolites derivatives and analogues. The compositions and methods disclosed herein are intended to be applied to any and all known derivatives or analogues of polyglutamine-oxidized folate antimetabolites. In some embodiments, the analogues correspond to modified forms of the folate antimetabolites, in which case the glutamyl group of the folate antimetabolites is not bound to the remainder of the folate antimetabolites molecule via a gamma-peptide bond. In some embodiments, the analogues are variants of the folate antimetabolites, in which case the glutamyl group in the folate antimetabolites is of the D type. In some embodiments, the polyglutamine-oxidized form of the folate antimetabolites, or the polyglutamine-oxidized folate antimetabolites analogues or derivatives, are not fluorinated.

[0147] In some embodiments, the folate antagonist is selected from the following: methotrexate derivatives containing an indoline ring and modified ornithine, methotrexate derivatives containing an indoline ring and modified glutamic acid, methotrexate derivatives containing an alkyl-substituted benzene ring C, methotrexate derivatives partially containing benzoxazine, methotrexate derivatives partially containing benzothiazine, 10-deazaaminopterin analogs, 5-deazaaminopterin methotrexate analogs, 5,10-dideazaaminopterin methotrexate analogs, and Dorin-containing methotrexate derivatives, lipophilic amide methotrexate derivatives, L-threo-(2S,4S)-4-fluoroglutamic acid-containing methotrexate analogs, DL-3,3-difluoroglutamic acid-containing methotrexate analogs, methotrexate tetrahydroquinazoline analogs, N-(ac-aminoacyl)methotrexate derivatives, biotin methotrexate derivatives, D-glutamic acid methotrexate analogs, D-erythro,threo-4-fluoroglutamic acid methotrexate analogs, β,γ-methano Methotrexate analogs, 10-deazaaminopterin (10-EDAM) analogs, γ-tetrazolemethotrexate analogs, N-(L-α-aminoacyl)methotrexate derivatives, meta-isomers of aminopterin, ortho-isomers of aminopterin, hydroxymethylmethotrexate, γ-fluoromethotrexate, polyglutamylmethotrexate derivatives, gem-diphosphonate-methotrexate analogs (see, for example, International Application No. 1988 / 06158, the contents of which are incorporated herein by reference in their entirety), α-substituted methotrexate analogs, γ-substituted methotrexate analogs, 5-methyl-5-deazamethotrexate analogs (see, for example, U.S. Patent No. 4,725,See issue 687, which is incorporated herein by reference in its entirety), N δ-acyl-N α-(4-amino-4-deoxypteroyl)-L-ornithine derivatives, 8-deazamethotrexate analogs, asibicinmethotrexate analogs, polymerplatinolmethotrexate derivatives, methotrexate-γ-dimyristoylphophatidylethanolamine, methotrexate polyglutamate analogs, poly-γ-glutamylmethotrexate derivatives, deoxyuridilatemethotrexate derivatives, iodoacetyllysinemethot Lexate analogs, 2,ω-diaminoalkanoid acid-containing methotrexate analogs, polyglutamate derivatives, 5-methyl-5-deaza analogs, quinazoline methotrexate analogs, pyrazine methotrexate analogs, cysteine ​​or homocysteine ​​methotrexate analogs (see, for example, U.S. Patent No. 4,490,529 and EPA Patent No. 0142220, the contents of which are incorporated herein by reference in their entirety), γ-tert- Butylmethotrexate ester, fluoride methotrexate analog, folic acid methotrexate analog, phosphonoglutamic acid analog, poly(L-lysine)methotrexate complex, dilysine or trilysine methotrexate derivatives, 7-hydroxymethotrexate, poly-γ-glutamylmethotrexate analog, 3',5'-dichloromethotrexate, diazoketone or chloromethyl ketone methotrexate analog, 10-propargyl Luaminopterin, alkylmethotrexate homologs, lectin derivatives of methotrexate, polyglutamate methotrexate derivatives, halogenated methotrexate derivatives, 8-alkyl-7,8-dihydro analogs, 7-methylmethotrexate derivatives, dichloromethotrexate, lipophilic methotrexate derivatives, 3',5'-dichloromethotrexate, deazamethopterin analogs, and MX068, or their stereoisomers.

[0148] In further embodiments, the gamma-polyglutamine oxoxide antimetabolitic derivative or analog has a variant polyglutamate chain. In some embodiments, the polyglutamate chain contains one or more native or synthetic residues other than glutamate. In some embodiments, the polyglutamate chain contains one or more glutamyl groups that do not contain an amide bond. In other embodiments, one or more glutamyl groups of the polyglutamate chain are derivatized.

[0149] B.γANTIFOL-PG synthesis The folate antimetabolite polyglutamate compositions provided herein are obtained by the following known synthetic methods in the art: folate antimetabolites (including different pharmaceutically acceptable salts or acids (e.g., disodium folate antimetabolite) and crystalline and amorphous forms) and procedures for synthesizing intermediates for the synthesis of folate antimetabolites, but are not limited to U.S. Patents No. 2,512,572; No. 3,892,801; No. 3,989,703; No. 4,057,548; No. 4,067,867; No. 4,079,056; Examples include Nos. 4,080,325; Nos. 4,106,488; Nos. 4,136,101; Nos. 4,224,446; Nos. 4,306,064; Nos. 4,374,987; Nos. 4,421,913; Nos. 4,558,690; Nos. 4,662,359; and Nos. 4,767,859; as well as those listed in Calvert, Semin. Oncol. 26:3-10 (1999).

[0150] The folate antimetabolite polyglutamate compositions provided herein are obtained by the following synthetic methods using readily available reagents and synthetic intermediates. The addition of glutamyl residues to the glutamyl residues of the folate antimetabolite can be carried out using synthetic methods known in the art. In some embodiments, glutamyl residues are sequentially added to the glutamyl residues of the folate antimetabolite. In further embodiments, the polyglutamate is added to the glutamyl residues of the folate antimetabolite using "click chemistry" or other bioconjugate chemistry methods known to those skilled in the art. Alternatively, a peptide of glutamyl residues of a desired length can be generated and added to a glutamyl residue-free precursor of pemetrexed. The peptide can be prepared using methods known in the art. In some embodiments, the initial glutamyl residue is conjugated to wanglesin, and additional glutamyl residues are sequentially added by solid-phase peptide synthesis using F-moc chemistry. After the final glutamyl residue is added, the pemetrexed precursor binds to the peptide, and the molecule is cleaved from the resin.

[0151] C. Gamma-polyglutamine oxoxide antimetabolitic complex Surprisingly, the inventors have found that polyglutamic oxidase antagonists, such as folate antimetabolites (γPANTIFOL), can form complexes with other compositions, including therapeutic agents containing cytotoxic compounds such as platinum compounds. Accordingly, in some embodiments, the Disclosure provides complexes of γPANTIFOL (e.g., γPANTIFOL as disclosed herein) with a therapeutic agent or a salt or acid thereof. In some embodiments, the Disclosure provides complexes of γPANTIFOL as described in any of items [1] to

[11] of the section on embodiments for carrying out the invention with a therapeutic agent or a salt or acid thereof. In some embodiments, the γPANTIFOL / complex comprises γPANTIFOL and a therapeutic agent. In some embodiments, the therapeutic agent is a cytotoxic compound such as a chemotherapeutic agent. In further embodiments, the γPANTIFOL / complex comprises a platinum-based drug such as a platinum-based chemotherapeutic agent (e.g., carboplatin and cisplatin). In other embodiments, the γPANTIFOL / complex comprises a taxane-based chemotherapeutic agent (e.g., carboplatin and cisplatin). In other embodiments, the γPANTIFOL / complex comprises a cyclodextrin. In further embodiments, the γPANTIFOL / complex is encapsulated within liposomes. In some embodiments, the liposomes are Lp-αPANTIFOL as described in any of items

[12] to

[67] .

[0152] In further embodiments, the γPANTIFOL / therapeutic complex comprises one or more γPANTIFOLs containing 2-150, 2-100, 2-75, 2-50, 2-24, 2-30, 2-20, 2-19, 2-15, 2-10, or 2-5 glutamyl groups. In some embodiments, the γPANTIFOL / therapeutic complex comprises one or more γPANTIFOLs containing 3-10, 3-9, 3-8, or 3-7 glutamyl groups, or any range in between. In other embodiments, the γPANTIFOL / therapeutic complex comprises one or more γPANTIFOLs containing 4-10, 4-9, 4-8, 4-7, 4-6, or 4-5 glutamyl groups, or any range in between. In a particular embodiment, the complex comprises one or more γPANTIFOLs containing 3-10 glutamyl groups. In further embodiments, the γPANTIFOL / therapeutic complex comprises one or more γPANTIFOLs containing 3 to 7 glutamyl groups. In another embodiment, the γPANTIFOL / therapeutic complex comprises one or more γPANTIFOLs containing 5 glutamyl groups. In yet another embodiment, the γPANTIFOL / therapeutic complex comprises one or more γPANTIFOLs containing 6 glutamyl groups. In some embodiments, the therapeutic agent is a cytotoxic compound or a salt or acid thereof. In further embodiments, the therapeutic agent is a chemotherapeutic agent or a salt or acid thereof. In another embodiment, the chemotherapeutic agent is a platinum-based drug. In yet another embodiment, the chemotherapeutic agent is a taxane-based drug. In further embodiments, the molar ratio of γPANTIFOL / therapeutic agent in the complex is in the range of 1 to 10:1. In some embodiments, the molar ratio of γPANTIFOL / therapeutic agent in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, or 10:1. In some embodiments, the γPANTIFOL / therapeutic complex is encapsulated in liposomes (for example, as described herein or by other methods known in the art).In some embodiments, the molar ratio of γPANTIFOL / therapeutic agent in the complex is 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:(21-50), or 1:>50. In some embodiments, the molar ratio of γPANTIFOL / therapeutic agent in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, (21-50):1, or >50:1. In some embodiments, the γPANTIFOL / therapeutic agent complex is encapsulated in liposomes (e.g., as described herein or by another method known in the art). In some embodiments, the liposomes are Lp-αPANTIFOL as described in any of items

[12] -

[67] of the section on embodiments for carrying out the invention.

[0153] In alternative embodiments, the γPANTIFOL complex comprises γPANTIFOL and cyclodextrin. In some embodiments, the γPANTIFOL complex comprises γPANTIFOL as described in any of items [1] to

[11] of the section on embodiments for carrying out the invention. In some embodiments, the γPANTIFOL complex comprises a folate antagonist as described in Section I. In some embodiments, the molar ratio of γPANTIFOL (e.g., γPANTIFOL salt) / cyclodextrin in the complex is in the range of 1 to 20:1 or any range in between. In some embodiments, the molar ratio of γPANTIFOL / cyclodextrin in the complex is in the range of 1 to 10:1 or any range in between. In further embodiments, the molar ratio of γPANTIFOL / cyclodextrin in the complex is in the range of 2 to 8:1 or any range in between. In some embodiments, the molar ratio of γPANTIFOL / cyclodextrin in the complex is 1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, or 20:1. In some embodiments, the molar ratio of γPANTIFOL / cyclodextrin in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, (21-50):1, or >50:1. In other embodiments, the molar ratio of γPANTIFOL / cyclodextrin in the complex is in the range of 1:1 to 20, 1:1 to 10, or 1:2 to 8, or any range in between. In some embodiments, the molar ratio of γPANTIFOL / cyclodextrin in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, or 1:20.In some embodiments, the molar ratio of γPANTIFOL / cyclodextrin in the complex is 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:(21-50), or 1:>50. In some embodiments, the γPANTIFOL / cyclodextrin complex is encapsulated in liposomes (e.g., as described herein or by another method known in the art). In some embodiments, the liposomes are Lp-γPANTIFOL as described in any of items

[12] -

[67] of the section on embodiments for carrying out the invention.

[0154] In some embodiments, the Disclosure provides compositions comprising a γPANTIFOL / platinum-based chemotherapeutic agent conjugate. In some embodiments, the conjugate comprises γPANTIFOL as described in any of items [1] to

[11] of the section on embodiments for carrying out the invention. In some embodiments, the γPANTIFOL conjugate comprises a polyglutamic acid oxidase antagonist as described in Section I. In some embodiments, the platinum-based chemotherapeutic agent is selected from cisplatin, carboplatin, and oxaliplatin, or their salts or acids. In other embodiments, the γPANTIFOL / platinum-based chemotherapeutic agent conjugate comprises cisplatin, carboplatin, an analogue of oxaliplatin, or their salts or acids. In some embodiments, the molar ratio of γPANTIFOL / platinum-based agent in the conjugate is in the range of 1 to 20:1, or any range in between. In some embodiments, the molar ratio of γPANTIFOL / platinum-based agent in the conjugate is in the range of 1 to 10:1, or any range in between. In further embodiments, the molar ratio of γPANTIFOL / platinum-based drug in the complex is in the range of 2 to 8:1, or any range in between. In some embodiments, the molar ratio of γPANTIFOL / platinum-based drug in the complex is 11:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, or 20:1. In some embodiments, the molar ratio of γPANTIFOL / platinum-based agent in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, (21-50):1, or >50:1. In other embodiments, the molar ratio of γPANTIFOL / platinum-based chemotherapeutic agent in the complex is in the range of 1:1-20, 1:1-10, or 1:2-8, or any range in between.In some embodiments, the molar ratio of γPANTIFOL / platinum-based drug in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, or 1:20. In some embodiments, the molar ratio of γPANTIFOL / platinum-based drug in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:(21~50), or 1:>50. In some embodiments, the γPANTIFOL / platinum-based drug conjugate is encapsulated in liposomes. In some embodiments, the liposomes are Lp-αPANTIFOL as described in any of the items

[12] to

[67] of the section on embodiments for carrying out the invention.

[0155] In further embodiments, the γPANTIFOL / platinum-based chemotherapeutic agent conjugate comprises an analog of cisplatin, carboplatin, oxaliplatin, or a salt or acid thereof. In some embodiments, the conjugate comprises γPANTIFOL as described in any of items [1] to

[11] of the section on embodiments for carrying out the invention. In some embodiments, the γPANTIFOL conjugate comprises a polyglutamic acid oxidase antagonist as described in Section I. In some embodiments, the molar ratio of γPANTIFOL / platinum-based analog in the conjugate is in the range of 1 to 20:1 or any range in between. In some embodiments, the molar ratio of γPANTIFOL / platinum-based analog in the conjugate is in the range of 1 to 10:1 or any range in between. In further embodiments, the molar ratio of γPANTIFOL / platinum-based agent in the conjugate is in the range of 2 to 8:1 or any range in between. In some embodiments, the molar ratio of γPANTIFOL / platinum-based analogs in the complex is 11:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, or 20:1. In some embodiments, the molar ratio of γPANTIFOL / platinum-based analogs in the complex is 11:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, (21-50):1, or >50:1. In some embodiments, the molar ratio of γPANTIFOL / platinum-based drug in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, or 1:20. In some embodiments, the molar ratio of γPANTIFOL / platinum-based drug in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:(21~50), or 1:>50.In some embodiments, the γPANTIFOL / platinum-based analog complex is encapsulated in liposomes. In some embodiments, the liposome is Lp-αPANTIFOL as described in any of items

[12] to

[67] of the section of forms for carrying out the invention.

[0156] In further embodiments, the Disclosure provides a complex comprising γPANTIFOL and cisplatin or a salt or acid thereof. In some embodiments, the complex comprises γPANTIFOL as described in any of items [1] to

[11] of the section on embodiments for carrying out the invention. In some embodiments, the γPANTIFOL complex comprises a folate antagonist as described in Section I. In some embodiments, the molar ratio of γPANTIFOL / cisplatin (or a salt or acid of cisplatin) in the complex is in the range of 1 to 20:1, or any range in between. In some embodiments, the molar ratio of γPANTIFOL / cisplatin (or a salt or acid of cisplatin) in the complex is in the range of 1 to 10:1, or any range in between. In further embodiments, the molar ratio of γPANTIFOL / cisplatin (or a salt or acid of cisplatin) in the complex is in the range of 2 to 8:1, or any range in between. In some embodiments, the molar ratio of γPANTIFOL / cisplatin (or a salt or acid of cisplatin) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, or 1:20. In some embodiments, the molar ratio of γPANTIFOL / cisplatin (or a salt or acid of cisplatin) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, (21-50):1, or >50:1. In some embodiments, the molar ratio of γPANTIFOL / cisplatin (or a salt or acid of cisplatin) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, or 1:20.In some embodiments, the molar ratio of γPANTIFOL / cisplatin (or a salt or acid of cisplatin) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:(21-50), or 1:>50. In some embodiments, the γPANTIFOL / cisplatin (or a salt or acid of cisplatin) complex is encapsulated in liposomes. In some embodiments, the liposomes are Lp-αPANTIFOL as described in any of the items

[12] -

[67] of the section on embodiments for carrying out the invention.

[0157] In other embodiments, the Disclosure provides a complex comprising γPANTIFOL and carboplatin or a salt or acid thereof. In some embodiments, the complex comprises γPANTIFOL as described in any of items [1] to

[11] of the section on embodiments for carrying out the invention. In some embodiments, the γPANTIFOL complex comprises a polyglutamic oxidase antimetabolite as described in Section I of this Spec. In some embodiments, the molar ratio of γPANTIFOL / carboplatin (or a salt or acid of carboplatin) in the complex is in the range of 1 to 20:1 or any range in between. In further embodiments, the molar ratio of γPANTIFOL / carboplatin (or a salt or acid of carboplatin) in the complex is in the range of 1 to 10:1 or any range in between. In further embodiments, the molar ratio of γPANTIFOL / carboplatin (or a salt or acid of carboplatin) in the complex is in the range of 2 to 8:1 or any range in between. In some embodiments, the molar ratio of γPANTIFOL / carboplatin (or a salt or acid of carboplatin) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, or 20:1. In some embodiments, the molar ratio of γPANTIFOL / carboplatin (or a salt or acid of carboplatin) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, (21-50):1, or >50:1. In some embodiments, the molar ratio of γPANTIFOL / carboplatin in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, or 1:20.In some embodiments, the molar ratio of γPANTIFOL / carboplatin in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:(21 to 50), or 1:>50. In some embodiments, the γPANTIFOL / carboplatin (or a salt or acid of carboplatin) complex is encapsulated in liposomes. In some embodiments, the liposomes are Lp-αPANTIFOL as described in any of items

[12] to

[67] in the section of forms for carrying out the invention.

[0158] In other embodiments, the Disclosure provides a complex comprising γPANTIFOL and oxaliplatin or a salt or acid thereof. In some embodiments, the complex comprises γPANTIFOL as described in any of the items [1] to

[11] of the section on embodiments for carrying out the invention. In some embodiments, the γPANTIFOL complex comprises a polyglutamic oxyl folate antimetabolite as described in Section I. In some embodiments, the molar ratio of γPANTIFOL / oxaliplatin (or a salt or acid of oxaliplatin) in the complex is in the range of 1 to 20:1 or any range in between. In further embodiments, the molar ratio of γPANTIFOL / oxaliplatin (or a salt or acid of oxaliplatin) in the complex is in the range of 1 to 10:1 or any range in between. In further embodiments, the molar ratio of γPANTIFOL / oxaliplatin (or a salt or acid of oxaliplatin) in the complex is in the range of 2 to 8:1 or any range in between. In some embodiments, the molar ratio of γPANTIFOL / oxaliplatin (or a salt or acid of oxaliplatin) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, or 1:20. In some embodiments, the molar ratio of γPANTIFOL / oxaliplatin (or a salt or acid of oxaliplatin) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, (21-50):1, or >50:1. In some embodiments, the molar ratio of γPANTIFOL / oxaliplatin (or a salt or acid of oxaliplatin) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, or 1:20.In some embodiments, the molar ratio of γPANTIFOL / oxaliplatin (or a salt or acid of oxaliplatin) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:(21-50), or 1:>50. In further embodiments, the γPANTIFOL / oxaliplatin (or a salt or acid of oxaliplatin) complex is encapsulated in liposomes. In some embodiments, the liposomes are Lp-αPANTIFOL as described in any of the items

[12] -

[67] of the section on embodiments for carrying out the invention.

[0159] In further embodiments, the disclosure provides a conjugate comprising γPANTIFOL and a platinum-based chemotherapeutic agent (platinum) selected from nedaplatin, heptaplatin, lovaplatin, stratoplatin, paraplatin, platinol, cycloplatin, dexormaplatin, spiroplatin, picoplatin, triplatin, tetraplatin, iproplatin, ormaplatin, zeniplatin, platinum-triamine, satoraplatin, enloplatin, JM216, NK121, CI973, DWA2114R, NDDP, and nedaplatin, or salts or acids thereof. In other embodiments, the γPANTIFOL / platinum-based chemotherapeutic agent conjugate includes nedaplatin, heptaplatin, lovaplatin, stratoplatin, paraplatin, platinol, cycloplatin, dexormaplatin, spiroplatin, picoplatin, triplatin, tetraplatin, iproplatin, ormaplatin, zeniplatin, platinum-triamine, satoraplatin, enloplatin, JM216, NK121, CI973, DWA2114R, NDDP, or analogues of nedaplatin, or salts or acids thereof. In some embodiments, the molar ratio of γPANTIFOL / platinum-based chemotherapeutic agent (platinum) (or salts or acids of platinum-based chemotherapeutic agents) in the conjugate is in the range of 1 to 20:1, or any range in between. In some embodiments, the conjugate includes γPANTIFOL as described in any of items [1] to

[11] of the section on embodiments for carrying out the invention. In some embodiments, the γPANTIFOL complex comprises a polyglutamic oxidase antimetabolite as described in Section I. In further embodiments, the molar ratio of γPANTIFOL / platinum (or a salt or acid of platinum) in the complex is in the range of 1 to 10:1, or any range in between. In further embodiments, the molar ratio of γPANTIFOL / platinum (or a salt or acid of platinum) in the complex is in the range of 2 to 8:1, or any range in between. In some embodiments, the molar ratio of γPANTIFOL / platinum (or a salt or acid of platinum) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, or 20:1.In some embodiments, the molar ratio of γPANTIFOL / platinum (or a platinum salt or acid) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, (21-50):1, or >50:1. In some embodiments, the molar ratio of γPANTIFOL / platinum (or a platinum salt or acid) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, or 1:20. In some embodiments, the molar ratio of γPANTIFOL / platinum (or a salt or acid of platinum) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:(21-50), or 1:>50. In some embodiments, the γPANTIFOL / platinum (or a salt or acid or analog thereof) complex is encapsulated in liposomes. In some embodiments, the liposomes are Lp-αPANTIFOL as described in any of the items

[12] -

[67] of the section on embodiments for carrying out the invention.

[0160] In some embodiments, the Disclosure provides compositions comprising a γPANTIFOL / taxane-based chemotherapeutic agent (taxane) conjugate. In some embodiments, the conjugate comprises a γPANTIFOL described in any of the items [1] to

[11] of the section on embodiments for carrying out the invention. In some embodiments, the γPANTIFOL conjugate comprises a polyglutamic acid oxidase antagonist described in Section I. In some embodiments, the taxane-based chemotherapeutic agent is selected from paclitaxel (PTX), docetaxel (DTX), larotaxel (LTX), and cabazitaxel (CTX), or their salts or acids. In some embodiments, the molar ratio of γPANTIFOL / taxane (or salt or acid of taxane) in the conjugate is in the range of 1 to 20:1, or any range in between. In further embodiments, the molar ratio of γPANTIFOL / taxane (or salt or acid of taxane) in the conjugate is in the range of 1 to 10:1, or any range in between. In further embodiments, the molar ratio of γPANTIFOL / taxane (or a salt or acid of a taxane) in the complex is in the range of 2 to 8:1, or any range in between. In some embodiments, the molar ratio of γPANTIFOL / taxane (or a salt or acid of a taxane) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, or 20:1. In some embodiments, the molar ratio of γPANTIFOL / taxane (or salt or acid of taxane) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, (21-50):1, or >50:1. In some embodiments, the molar ratio of γPANTIFOL / taxane (or salt or acid of taxane) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, or 1:20.In some embodiments, the molar ratio of γPANTIFOL / taxane (or salt or acid of taxane) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:(21-50), or 1:>50. In further embodiments, the γPANTIFOL / taxane (or salt or acid of taxane) drug complex is encapsulated in liposomes. In some embodiments, the liposomes are Lp-αPANTIFOL as described in any of the items

[12] -

[67] of the section on embodiments for carrying out the invention.

[0161] In further embodiments, the Disclosure provides a complex comprising γPANTIFOL and paclitaxel (PTX) or a salt or acid thereof. In other embodiments, the γPANTIFOL / paclitaxel (or a salt or acid of paclitaxel) chemotherapeutic agent complex comprises an analogue of paclitaxel (PTX), or a salt or acid thereof. In some embodiments, the complex comprises γPANTIFOL as described in any of items [1] to

[11] of the section on embodiments for carrying out the invention. In some embodiments, the γPANTIFOL complex comprises a polyglutamic oxidase antimetabolite as described in Section I. In some embodiments, the molar ratio of γPANTIFOL / paclitaxel (or a salt or acid of paclitaxel) in the complex is in the range of 1 to 20:1, or any range in between. In further embodiments, the molar ratio of γPANTIFOL / paclitaxel (or a salt or acid of paclitaxel) in the complex is in the range of 1 to 10:1, or any range in between. In further embodiments, the molar ratio of γPANTIFOL / paclitaxel (or a salt or acid of paclitaxel) in the complex is in the range of 2 to 8:1, or any range in between. In some embodiments, the molar ratio of γPANTIFOL / paclitaxel (or a salt or acid of paclitaxel) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, or 20:1. In some embodiments, the molar ratio of γPANTIFOL / paclitaxel (or a salt or acid of paclitaxel) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, (21-50):1, or >50:1.In some embodiments, the molar ratio of γPANTIFOL / paclitaxel (or a salt or acid of paclitaxel) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, or 1:20. In some embodiments, the molar ratio of γPANTIFOL / paclitaxel (or a salt or acid of paclitaxel) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:(21-50), or 1:>50. In further embodiments, the γPANTIFOL / paclitaxel (or a salt or acid of paclitaxel) complex is encapsulated in liposomes. In some embodiments, the liposomes are Lp-αPANTIFOL as described in any of the items

[12] -

[67] of the section on embodiments for carrying out the invention.

[0162] In further embodiments, the Disclosure provides a complex comprising γPANTIFOL and docetaxel (DTX) or a salt or acid thereof. In other embodiments, the γPANTIFOL / docetaxel complex comprises an analog of docetaxel (DTX) or a salt or acid thereof. In some embodiments, the complex comprises γPANTIFOL as described in any of items [1] to

[11] of the section on embodiments for carrying out the invention. In some embodiments, the γPANTIFOL complex comprises a polyglutamic oxidase antimetabolite as described in Section I. In some embodiments, the molar ratio of γPANTIFOL / docetaxel (or a salt or acid of docetaxel) in the complex is in the range of 1 to 20:1 or any range in between. In some embodiments, the molar ratio of γPANTIFOL / docetaxel (or a salt or acid of docetaxel) in the complex is in the range of 1 to 10:1 or any range in between. In further embodiments, the molar ratio of γPANTIFOL / docetaxel (or a salt or acid of docetaxel) in the complex is in the range of 2 to 8:1, or any range in between. In some embodiments, the molar ratio of γPANTIFOL / docetaxel (or a salt or acid of docetaxel) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, or 20:1. In some embodiments, the molar ratio of γPANTIFOL / docetaxel (or a salt or acid of docetaxel) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, (21-50):1, or >50:1. In some embodiments, the molar ratio of γPANTIFOL / docetaxel (or a salt or acid of docetaxel) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, or 1:20.In some embodiments, the molar ratio of γPANTIFOL / docetaxel (or a salt or acid of docetaxel) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:(21-50), or 1:>50. In further embodiments, the γPANTIFOL / docetaxel (or a salt or acid of docetaxel) complex is encapsulated in liposomes. In some embodiments, the liposomes are Lp-αPANTIFOL as described in any of the items

[12] -

[67] of the section on embodiments for carrying out the invention.

[0163] In further embodiments, the Disclosure provides a complex comprising γPANTIFOL and larotaxel (LTX) or a salt or acid thereof. In some embodiments, the complex comprises γPANTIFOL as described in any of items [1] to

[11] of the section on embodiments for carrying out the invention. In some embodiments, the γPANTIFOL complex comprises a polyglutamic oxidase antimetabolite as described in Section I. In some embodiments, the molar ratio of γPANTIFOL / larotaxel (or a salt or acid of larotaxel) in the complex is in the range of 1 to 20:1 or any range in between. In further embodiments, the molar ratio of γPANTIFOL / larotaxel (or a salt or acid of larotaxel) in the complex is in the range of 1 to 10:1 or any range in between. In further embodiments, the molar ratio of γPANTIFOL / larotaxel (or a salt or acid of larotaxel) in the complex is in the range of 2 to 8:1 or any range in between. In some embodiments, the molar ratio of γPANTIFOL / larotaxel (or a salt or acid of larotaxel) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, or 20:1. In some embodiments, the molar ratio of γPANTIFOL / larotaxel (or a salt or acid of larotaxel) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, (21-50):1, or >50:1. In some embodiments, the molar ratio of γPANTIFOL / larotaxel (or a salt or acid of larotaxel) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, or 1:20.In some embodiments, the molar ratio of γPANTIFOL / larotaxel (or a salt or acid of larotaxel) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:(21-50), or 1:>50. In further embodiments, the γPANTIFOL / larotaxel (or a salt or acid of larotaxel) complex is encapsulated in liposomes. In some embodiments, the liposomes are Lp-αPANTIFOL as described in any of the items

[12] -

[67] of the section on embodiments for carrying out the invention.

[0164] In further embodiments, the Disclosure provides a complex comprising γPANTIFOL and cabazitaxel (CTX) or a salt or acid thereof. In some embodiments, the complex comprises γPANTIFOL as described in any of items [1] to

[11] of the section on embodiments for carrying out the invention. In some embodiments, the γPANTIFOL complex comprises a polyglutamic oxidase antagonist as described in Section I. In some embodiments, the molar ratio of γPANTIFOL / cabazitaxel (or a salt or acid of cabazitaxel) in the complex is in the range of 1 to 20:1 or any range in between. In further embodiments, the molar ratio of γPANTIFOL / cabazitaxel (or a salt or acid of cabazitaxel) in the complex is in the range of 1 to 10:1 or any range in between. In further embodiments, the molar ratio of γPANTIFOL / cabazitaxel (or a salt or acid of cabazitaxel) in the complex is in the range of 2 to 8:1 or any range in between. In some embodiments, the molar ratio of γPANTIFOL / cabazitaxel (or a salt or acid of cabazitaxel) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, or 20:1. In some embodiments, the molar ratio of γPANTIFOL / cabazitaxel (or a salt or acid of cabazitaxel) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, (21-50):1, or >50:1. In some embodiments, the molar ratio of γPANTIFOL / cabazitaxel (or a salt or acid of cabazitaxel) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, or 1:20.In some embodiments, the molar ratio of γPANTIFOL / cabazitaxel (or a salt or acid of cabazitaxel) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:(21-50), or 1:>50. In further embodiments, the γPANTIFOL / cabazitaxel (or a salt or acid of cabazitaxel) complex is encapsulated in liposomes. In some embodiments, the liposomes are Lp-αPANTIFOL as described in any of the items

[12] -

[67] of the section on embodiments for carrying out the invention.

[0165] In further embodiments, the Disclosure provides a complex comprising γPANTIFOL and another antimetabolite, or a salt or acid thereof. In some embodiments, the complex comprises γPANTIFOL as described in any of items [1] to

[11] of the section on embodiments for carrying out the invention. In some embodiments, the γPANTIFOL complex comprises a polyglutamic oxidase folate antimetabolite as described in Section I. An antimetabolite is a chemical substance that is similar in structure to a metabolite required for normal biochemical reactions but is sufficiently different to interfere with the normal function of one or more cells, such as cell division. In some embodiments, the Disclosure provides a complex comprising γPANTIFOL and a folate antimetabolite (ANTIFOL), or a salt or acid thereof. In some embodiments, the disclosure provides a complex comprising γPANTIFOL and an antimetabolite selected from gemcitabine, fluorouracil, capecitabine, antiphorates (e.g., folic acid antimetabolites, larcitrexed), tegafur, cytosine arabinoside, thioguanine, 5-azacitidine, 6-mercaptopurine, azathioprine, 6-thioguanine, pentostatin, fludarabine phosphate, and cladribine, as well as any pharmaceutically acceptable salt or acid (one or more) or derivative thereof. In some embodiments, the molar ratio of γPANTIFOL / antimetabolite (or salt or acid of an antimetabolite, or prodrug) in the complex is in the range of 1 to 20:1 or any range in between. In further embodiments, the molar ratio of γPANTIFOL / antimetabolite (or salt or acid of an antimetabolite, or prodrug) in the complex is in the range of 1 to 10:1 or any range in between. In further embodiments, the molar ratio of γPANTIFOL / antimetabolite (or salt or acid of the antimetabolite, or prodrug) in the complex is in the range of 2 to 8:1, or any range in between.In some embodiments, the molar ratio of γPANTIFOL / antimetabolite (or salt or acid of antimetabolite, or prodrug) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, or 20:1. In some embodiments, the molar ratio of γPANTIFOL / antimetabolite (or salt or acid of the antimetabolite, or prodrug) in the complex is 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, (21-50):1, or >50:1. In some embodiments, the molar ratio of γPANTIFOL / antimetabolite (or salt or acid of antimetabolite, or prodrug) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, or 1:20. In some embodiments, the molar ratio of γPANTIFOL / antimetabolite (or salt or acid of the antimetabolite, or prodrug) in the complex is 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:(21-50), or 1:>50. In further embodiments, the γPANTIFOL / antimetabolite (or salt or acid of the antimetabolite, or prodrug) complex is encapsulated in liposomes. In some embodiments, the liposomes are Lp-αPANTIFOL as described in any of the items

[12] -

[67] of the section on embodiments for carrying out the invention.

[0166] In further embodiments, the Disclosure provides complexes of γPANTIFOL (e.g., γPANTIFOL as disclosed herein) and cyclodextrins. Cyclodextrins (CDs) are a group of cyclic oligosaccharides that have been shown to improve the physicochemical properties of many drugs through the formation of complexes. CDs are cyclic oligosaccharides composed of several D-glucose units linked by α-(1,4) bonds. This cyclic structure provides a hydrophobic internal cavity, giving CDs a shortened conical shape. Many hydroxyl groups are located on the ends of the ring, which makes CDs both lipophilic and water-soluble. As a result, CDs can form complexes with a wide variety of hydrophobic drugs, thereby altering the physicochemical properties of these complexed drugs. In some embodiments, the complexes include γPANTIFOL as described in any of items [1] to

[11] of the section on embodiments for carrying out the invention.

[0167] The term "cyclodextrin" or "CD" usually refers to a parent or derivatized cyclic oligosaccharide that can form a complex with folate antimetabolite-prostaglandins (PGs), unless otherwise specified, and containing a variable number of (α-1,4)-linked D-glucopyranoside units. Each cyclodextrin glucopyranoside subunit has a secondary hydroxyl group at positions 2 and 3 and a primary hydroxyl group at position 6. The terms "parent," "non-derivativeized," or "inactive" cyclodextrins contain a D-glucopyranoside unit and have the basic formula C6H 12 This refers to cyclodextrins that have an O6 and glucose structure and no additional chemical substituents (e.g., α-cyclodextrin consisting of 6 D-glucopyranoside units, β-cyclodextrin consisting of 7 D-glucopyranoside units, and γ-cyclodextrin consisting of 8 D-glucopyranoside units). The physical and chemical properties of parent cyclodextrins can be modified by derivatizing the hydroxyl group with other functional groups. Any substance located in the cyclodextrin internal phase is said to be "complexed" with the cyclodextrin, or to form a complex (inclusion complex) with the cyclodextrin.

[0168] As used herein, there are no special restrictions on the cyclodextrin component of the γPANTIFOL / cyclodextrin complex, as long as the cyclodextrin can form a complex with γPANTIFOL. In certain embodiments, the cyclodextrin is derivatized to have an ionizable (e.g., weakly basic and / or weakly acidic) functional group to facilitate complex formation with γPANTIFOL and / or liposome encapsulants.

[0169] Modification of hydroxyl groups of cyclodextrins, such as hydroxyl groups directed away from the cyclodextrin internal phase using ionizable chemical groups, is known to facilitate the addition of cyclodextrins and therapeutic agents complexed with cyclodextrins. In some embodiments, the cyclodextrin in the γPANTIFOL / cyclodextrin complex has hydroxyl groups substituted with at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 ionizable chemical groups. The term “charged cyclodextrin” means a cyclodextrin having hydroxyl groups substituted with one or more of its charged moieties. Such moieties may be charged groups themselves or may include organic moieties (e.g., C1-C6 alkyl or C1-C6 alkyl ether moieties) substituted with one or more charged moieties.

[0170] In some embodiments, the "ionizable" or "charged" portion of the CD derivative is weakly ionizable. The weakly ionizable portion is either a weakly basic or weakly acidic portion. The weakly basic functional group (W) has a pKa of CH3-W in the range of approximately 6.0–9.0, 6.5–8.5, 7.0–8.0, 7.5–8.0, and any range in between (including endpoints). Similarly, the weakly acidic functional group (X) has a logged dissociation constant (pKa) of CH3-X in the range of approximately 3.0–7.0, 4.0–6.5, 4.5–6.5, 5.0–6.0, 5.0–5.5, and any range in between (including endpoints). Typical anionic moieties include, but are not limited to, carboxylates, carboxymethyls, succinyls, sulfonyls, phosphates, sulfoalkyl ethers, sulfate carbonates, thiocarbonates, dithiocarbonates, phosphates, phosphonates, sulfonates, nitrates, and borate groups. Typical cationic moieties include, but are not limited to, aminos, guanidines, and quaternary ammonium groups.

[0171] In another embodiment, the derivatized cyclodextrin is a "polyanion" or a "polycation." A polyanion is a derivatized cyclodextrin having two or more negatively charged groups, resulting in a net negative ionic charge of three or more units. A polycation is a derivatized cyclodextrin having two or more positively charged groups, resulting in a net positive ionic charge of three or more units.

[0172] In another embodiment, the derivatized cyclodextrin is a “chargeable amphiphilic substance.” “Chargeable” means that the amphiphilic substance has a pK in the range of pH 4 to pH 8 or 8.5. The charged amphiphilic substance can therefore be a weak acid or a base. As used herein, “amphoteric” means a derivatized cyclodextrin having ionizable groups with both anionic and cationic properties, wherein (a) at least one of the optionally cationic and anionic amphiphilic substances has at least one charge group having a pK between 4 and 8 to 8.5, (b) cationic charge is dominant at pH 4, and (c) anionic charge is dominant at pH 8 to 8.5.

[0173] In some embodiments, “ionizable” or “charged” derivatized cyclodextrins are generally weakly ionizable, whether polyionic, amphiphilic, or otherwise (i.e., having pKai of about 4.0–8.5, 4.5–8.0, 5.0–7.5, 5.5–7.5, 6.0–6.5, and any range in between (including both ends)).

[0174] Any one, some, or all of the hydroxyl groups of the α-D-glucopyranoside units of any one, some, or all of the cyclodextrins can be modified into ionizable chemical groups as described herein. Since each cyclodextrin hydroxyl group has a different chemical reactivity, the reaction with the modified portion can produce an amorphous mixture of positional and optical isomers. Alternatively, specific chemistry can be used to react the groups to form a homogeneous product of the pre-modified α-D-glucopyranoside units.

[0175] Aggregate substitution in cyclodextrin derivatives in a mixture is described using a term called the degree of substitution. For example, 6-ethylenediamino-β-cyclodextrin with a degree of substitution of 7 would consist of isomers of 6-ethylenediamino-β-cyclodextrin, each having 7 ethylenediamino groups per molecule. The degree of substitution in a mixture of cyclodextrin derivatives can be measured routinely using mass spectrometry or nuclear magnetic resonance spectroscopy.

[0176] In one embodiment, at least one hydroxyl group facing in the opposite direction from within the cyclodextrin is substituted with an ionizable chemical group. For example, at least one α-D-glucopyranoside unit among all three of the C2, C3, C6, C2 and C3, C2 and C6, C3 and C6, and C2-C3-C6 hydroxyls is substituted with an ionizable chemical group. Any such hydroxyl combination can be combined with any degree of substitution described herein, and similarly, at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and up to all α-D-glucopyranoside units in the modified cyclodextrin can be combined. One such derivative is sulfoalkyl ether cyclodextrin (SAE-CD). Sulfobutyl ether derivatives of beta-cyclodextrin (SBE-β-CD) have been shown to have significantly improved water solubility compared to the parent cyclodextrin.

[0177] Additional cyclodextrin derivatives that can complex with the therapeutic agent in the disclosed liposome compositions include sugammadex or Org-25969, in which case the 6-hydroxy group on γ-CD is substituted by a carboxythioacetate ether bond and hydroxybutenyl-β-CD. Alternative forms of cyclodextrin include 2,6-di-O-methyl-β-CD (DIMEB), 2-hydroxylpropyl-3-cyclodextrin (HP-β-CD), randomly methylated-β-cyclodextrin (RAMEB), sulfobutyl ether β-cyclodextrin (SBE-β-CD), and sulfobutyl ether-γ-cyclodextrin (SBEγCD), sulfobutylated-β-cyclodextrin sodium salt, (2-hydroxypropyl)-alpha-cyclodextrin, (2-hydroxypropyl)-β-cyclodextrin, (2-hydroxypropyl)-γ-cyclodextrin, 2,6-di-O-methyl)-β-cyclodextrin (DIMEB-50 heptakis), 2,3,6-tri-O-methyl)-β-cyclodextrin (TRIMEB heptakis), methyl-β-cyclodextrin, octakis(6-deoxy-6-iodo)-γ-cyclodextrin, and octakis(6-deoxy-6-bromo)-γ-cyclodextrin.

[0178] In some embodiments, the cyclodextrin has high solubility in water to facilitate capture of a greater amount of cyclodextrin in the inner phase of the liposome. In some embodiments, the aqueous solubility of the cyclodextrin is at least 10 mg / mL, 20 mg / mL, 30 mg / mL, 40 mg / mL, 50 mg / mL, 60 mg / mL, 70 mg / mL, 80 mg / mL, 90 mg / mL, 100 mg / mL, or more. In some embodiments, the water solubility of the cyclodextrin is in the range of 10-150 mg / mL, 20-100 mg / mL, 20-75 mg / mL, and within any range (including both ends) between these.

[0179] In some embodiments, a large binding constant between cyclodextrin and γPANTIFOL and / or other therapeutic agents complexed with cyclodextrin is preferred, which can be obtained by selecting the number of glucose units in the cyclodextrin based on the size of the therapeutic agent (e.g., Albers et al., Crit. Rev. Therap. Drug Carrier Syst. 12:311-337 (1995); Stella et al. See al., Toxicol. Pathol. 36:30-42 (2008). If the binding constant is pH-dependent, cyclodextrins can be selected such that the binding constant is greater at the pH of the liposome intraphase. As a result, the solubility (nominal solubility) of the therapeutic agent in the presence of cyclodextrin can be further improved. In some embodiments, the binding constants between cyclodextrin and therapeutic agent are 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000 or higher. In some embodiments, the binding constants between cyclodextrin and therapeutic agent are 100-1,200, 200-1,000, 300-750, and any range in between.

[0180] In some embodiments, the cyclodextrin in the γPANTIFOL / cyclodextrin complex and / or the cyclodextrin / therapeutic complex is not derivatized.

[0181] In some embodiments, the cyclodextrin in the γPANTIFOL / cyclodextrin complex and / or the cyclodextrin / therapeutic complex is derivatized. In further embodiments, the cyclodextrin derivative of the complex is of formula I: [ka] It has the structure, where n is 4, 5, or 6; R1, R2, R3, R4, R5, R6, R7, R8, and R9 are each independently -H, a linear or branched C1-C8 alkylene group, or an optionally substituted linear or branched C1-C6 group, and at least one of R1, R2, R3, R4, R5, R6, R7, R8, and R9 is a linear or branched C1-C8 alkylene (e.g., C1-C8-(alkylene)-SO3) - It is the basis.

[0182] In some embodiments, cyclodextrin derivatization of γPANTIFOL / cyclodextrin complexes and / or cyclodextrin / therapeutic complexes is performed using formula II: [ka] It has the structure, where n is 4, 5, or 6; R1, R2, R3, R4, R5, R6, R7, R8, and R9 are each independently -O- or -O-(C2-C6 alkylene)-SO3 - The group is; at least one of R1 and R2 is independently -O-(C2-C6 alkylene)-SO3 - The group is; S1, S2, S3, S4, S5, S6, S7, S8, and S9 are each independently pharmaceutically acceptable cations. In further embodiments, the pharmaceutically acceptable cations are Li + kaNa + , or K + Alkali metals such as Ca 2+ , or Mg 2+ Alkaline earth metals such as ammonium ions and amine cations such as (C1-C6)-alkylamines, piperidines, pyrazines, (C1-C6)-alkanolamines and (C4-C8)-cycloalkanolamines are selected. In some embodiments, at least one of R1 and R2 is independently -O-(CH2) mThe SO3- group is an -O-(C2-C6 alkylene)-SO3- group, where m is 2 to 6, preferably 2 to 4 (e.g., -O-CH2CH2CH2SO3- or -O-CH2CH2CH2CH2SO3-); and S1, S2, S3, S4, S5, S6, S7, S8, and S9 are each independently H or a pharmaceutically acceptable cation, which includes, for example, alkali metals (e.g., Li + kaNa + , K + ), alkaline earth metals (e.g., Ca 2+ Mg 2+ Examples include ammonium ions and amine cations such as (C1-C6)-alkylamines, piperidines, pyrazines, (C1-C6)-alkanolamines, and (C4-C8)-cycloalkanolamines.

[0183] In some embodiments, the cyclodextrin derivatization of γPANTIFOL / cyclodextrin complexes and / or cyclodextrin / therapeutic complexes is of the cyclodextrins disclosed in U.S. Patents 6,133,248, 5,874,418, 6,046,177, 5,376,645, 5,134,127, 7,034,013, and 6,869,939; and International Publication 02005 / 117911. The contents of each of these patent documents are incorporated herein by reference in a preferential manner.

[0184] In some embodiments, the cyclodextrin derivative of the γPANTIFOL / cyclodextrin complex and / or cyclodextrin / therapeutic complex is a sulfoalkyl ether cyclodextrin. In some embodiments, the cyclodextrin derivative of the complex is a sulfobutyl ether-3-cyclodextrin such as CAPTISOL® (CyDex Pharma, Inc., Lenexa, Kansas). Methods for producing sulfobutyl ether-3-cyclodextrin and other sulfoalkyl ether cyclodextrins are known in the art.

[0185] In some embodiments, the cyclodextrin derivative of the γPANTIFOL / cyclodextrin complex and / or cyclodextrin / therapeutic complex is of formula III: [ka] It is a compound of the form, where R is (a)(H) 21-X Or (-(CH2)4-SO3Na) X , and x = 1.0 to 10.0, 1.0 to 5.0, 6.0 to 7.0, or 8.0 to 10.0; (b)(H) 21-X Or (-(CH2CH(OH)CH3) X , and x = 1.0 to 10.0, 1.0 to 5.0, 6.0 to 7.0, or 8.0 to 10.0; (c)(H) 21-X or (sulfoalkyl ether) X , and x = 1.0~10.0, 1.0~5.0, 6.0~7.0, or 8.0~10.0; or (d)(H) 21-X Or (-(CH2)4-SO3Na) X , and x = 1.0 to 10.0, 1.0 to 5.0, 6.0 to 7.0, or 8.0 to 10.0.

[0186] In further embodiments, the γPANTIFOL / cyclodextrin complex and / or the cyclodextrin / therapeutic complex are encapsulated in liposomes (for example, as described herein or by other methods known in the art).

[0187] D.γPANTIFOL delivery carrier In alternative embodiments, the Disclosure provides γPANTIFOL delivery systems and their use for delivering γPANTIFOL payloads to cells (one or more) in vitro or in vivo. In some embodiments, γPANTIFOL is complexed with or incorporated into a delivery carrier. Such delivery carriers are known in the Art and are not limited to, but include liposomes, lipospheres, polymers, peptides, proteins, antibodies (e.g., ADCs such as antibody-γPANTIFOL complexes), cellular components, cyclic oligosaccharides (e.g., cyclodextrins), nanoparticles (e.g., lipid nanoparticles, biodegradable nanoparticles, and core-shell nanoparticles), lipoprotein particles, and combinations thereof. In certain embodiments, the delivery carrier is a liposome. In other specific embodiments, the delivery carrier is an antibody or antigen-binding antibody fragment. In some embodiments, the γPANTIFOL delivery system includes γPANTIFOL as described in any of items [1] to

[11] of the section on embodiments for carrying out the invention.

[0188] E. liposomes In some embodiments, the Disclosure provides liposome compositions comprising liposomes encapsulating (filled) a gamma-polyglutamate oxidase antagonist (e.g., γPANTIFOL as disclosed herein). In some embodiments, the liposome composition comprises γPANTIFOL as described in any of items [1] to

[11] of the section on embodiments for carrying out the invention. In some embodiments, the liposome composition comprises a polyglutamate oxidase antagonist as described in Section I. In some embodiments, the liposome composition comprises liposomes as described in any of items

[12] to

[67] of the section on embodiments for carrying out the invention. In some embodiments, the liposomes in the liposome composition comprise γPANTIFOL comprising 4, 5, 2 to 10, 4 to 6, or more than 5 glutamyl groups (including the glutamyl groups of the folate antagonist). In some embodiments, the gamma-polyglutamate oxidase antagonist in Lp-γPANTIFOL comprises 2 or more L-type glutamyl groups. In other embodiments, the gamma-polyglutamine oxidase antagonist in Lp-γPANTIFOL contains a D-type glutamyl group. In further embodiments, the gamma-polyglutamine oxidase antagonist in Lp-γPANTIFOL contains a D-type glutamyl group and two or more L-type glutamyl groups. In further embodiments, the gamma-polyglutamine oxidase antagonist in Lp-γPANTIFOL contains two or more glutamyl groups having a gamma-carboxyl bond. In some embodiments, the liposome composition contains liposomes containing a γ-pentaglutamine oxidase antagonist. In further embodiments, the liposomes contain an L-γ-pentaglutamine oxidase antagonist, a D-γ-pentaglutamine oxidase antagonist, or L- and D-γ-pentaglutamine oxidase antagonists. In some embodiments, the liposome composition comprises liposomes containing a γ-hexaglutamine oxidase antagonist (Lp-γPANTIFOL). In further embodiments, the liposomes comprise an L-γ-hexaglutamine oxidase antagonist, a D-γ-hexaglutamine oxidase antagonist, or L- and D-γ-hexaglutamine oxidase antagonists.In some embodiments, the liposome composition comprises anionic or neutral liposomes. In some embodiments, the liposome composition comprises cationic liposomes. In some embodiments, the Lp-γPANTIFOL composition is not pegylated. In some embodiments, the Lp-γPANTIFOL composition is not targeted (NTLp-γPANTIFOL). In some embodiments, the Lp-γPANTIFOL composition is targeted (TLp-γPANTIFOL). In some embodiments, the liposome composition comprises liposomes having a diameter of 20 nm to 500 nm, or any range in between. In some embodiments, the liposome composition comprises liposomes having a diameter of 20 nm to 400 nm, or any range in between. In some embodiments, the liposome composition comprises liposomes having a diameter of 20 nm to 200 nm, or any range in between. In further embodiments, the liposome composition comprises liposomes having a diameter of 20 nm to 150 nm, or any range in between. In further embodiments, the liposome composition comprises liposomes having a diameter of 80 nm to 120 nm, or any range in between. In further embodiments, 30 to 70%, 30 to 60%, or 30 to 50% w / w of gamma polyglutamate oxidase antagonist, or any range in between, is encapsulated in Lp-γPANTIFOL during the liposome preparation process. In some embodiments, the Lp-γPANTIFOL composition comprises at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or more than 75% w / w of gamma polyglutamate oxidase antagonist. In some embodiments, at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or more than 75% w / w of gamma polyglutamate oxidase folate antagonist is encapsulated in Lp-γPANTIFOL during the liposome preparation process.

[0189] In some embodiments, the provided liposomes further include an immunostimulant, a detectable marker, or both, disposed on the outer surface of the liposome. The immunostimulant or detectable marker can be ionically or covalently bonded to the outer surface of the liposome, and in this case, optionally, this may include binding to the steric-stabilizing components of the liposome.

[0190] The term "immunostimulatory agent," also known as "immunostimulant" or "immunostimulator," refers to a substance that stimulates immunity (including pre-existing immune responses) by inducing the activation or increased activity of any component of the immune system. These immunostimulators include one or more of the following: haptens, adjuvants, protein immunostimulants, nucleic acid immunostimulants, and chemoimmunostimulants. Many adjuvants contain substances designed to stimulate the immune response, such as lipid A, Bordetella pertussis, or Mycobacterium tuberculosis proteins. Certain adjuvants are commercially available, for example, Freund's incomplete and complete adjuvants (Difco Laboratories, Detroit, Mich.); Merck adjuvant 65 (Merck and Company, Inc., Rahway, NJ); AS-2 (SmithKline Beecham, Philadelphia, Pa.); aluminum salts such as aluminum hydroxide gel (alum) or aluminum phosphate; salts of calcium, iron, or zinc; insoluble suspensions of acylated tyrosine; acylated sugars; polysaccharides derivatized with cationic or anionic agents; polyphosphazenes; biodegradable microspheres; monophosphoryl lipid A and quil A; IFN alpha, IFN gamma, FLT3 ligands; and immunostimulant antibodies (e.g., anti-CTLA-4, anti-CD28, anti-CD3). Cytokines such as GM-CSF, interleukin 2, 7, 12, and 15, and other similar growth factors can also be used as adjuvants. In a preferred embodiment, the immunostimulant may be at least one selected from fluorescein, DNP, beta-glucan, beta-1,3-glucan, and beta-1,6-glucan. In a further preferred embodiment, the immunostimulant is a Toll-like receptor (TLR) modulator. In a further embodiment, the Toll-like receptor (TLR) modulator may be one or more of OXPAC, PGPC, erythraminophen (e.g., E5564), and resolvin.

[0191] In some embodiments, the provided liposomes further include a drug that increases the uptake of the liposome into the target intracellular compartment, including the cytosol. In some embodiments, the drug imparts the ability of the liposome contents to bypass lysosomes (e.g., chloroquine). In some embodiments, the drug improves the renewal of liposome contents by mitochondria (e.g., sphingomyelin and components of mitoport).

[0192] Examples of detectable markers include radioisotopes, fluorescent compounds, bioluminescent compounds, chemiluminescent compounds, metal chelators, enzymes, dyes, inks, magnetic compounds, biocatalysts, or pigments that are detectable by any suitable means known in the art, such as at least magnetic resonance imaging (MRI), optical imaging, fluorescence / emission imaging, or nuclear imaging techniques.

[0193] In some embodiments, the immunostimulant and / or detectable marker is bound to the outer surface of the liposome by co-incubation with the liposome. For example, the immunostimulant and / or detectable marker may be bound to the liposome membrane by hydrophobic interactions or ionic bonds, such as avidin / biotin bonds or metal chelate bonds (e.g., Ni-NTA). Alternatively, the immunostimulant or detectable marker may be covalently bound to the outer surface of the liposome, for example, by covalent bonding to a liposome component or to a steric stabilizer such as PEG.

[0194] One example of a reagent is fluorescein isothiocyanate (FITC), which, based on the inventors' experiments, surprisingly functions as both an immunostimulant and a detectable marker.

[0195] In some embodiments, the liposomes further include a drug that increases the uptake of the liposomes into the target intracellular compartment, incl...

Claims

1. A liposome composition comprising a gamma-polyglutamine-oxidized folate antimetabolite and one or more polyglutamine-oxidized or polyglutamine-oxidized folate antimetabolite or polyglutamine-oxidized folate antimetabolite, (a) The gamma-polyglutamine oxoxide antimetabolitic agent comprises a polyglutamate chain containing 2 to 15 glutamyl groups, wherein at least one glutamyl group is linked to another glutamyl group via a gamma-carboxyl group bond; (b) The polyglutamine oxidase antagonist is pralatrexate, AG2034, GW1843, and LY309887, or their stereoisomers; RTX and LMX, or their stereoisomers; 2-dMTX, 2-desamino-MTX; 2-CH3-MTX, 2-desamino-2-methyl-MTX; AMT, aminopterin; 2-dAMT, 2-desamino-AMT; 2-CH3-AMT, 2-desamino -2-methyl-AMT;10-EdAM,10-ethyl-10-deazaaminopterin;DDATHF(lometrexol),5,10-dideaza-5,6,7,8-tetrahydrofolate;5-d(i)H4PteGlu,5-deaza-5,6,7,8-tetrahydroisofolate;N9-CH3-5-d(i)H4PteGlu,N9-methyl-5-deaza-5,6,7,8-tetrahydroisofolate;5-dPteHCysA,N Alpha-(5-deazapteroyl)-L-homocysteic acid; 5-dPteAPBA,N Alpha-(5-deazapteroyl)-DL-2-amino-4-phosphonobutyric acid; 5-dPteOrn,N Alpha-(5-deazapteroyl)-L-ornithine; 5-dH4PteHCysA,N Alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-L-homocysteic acid; 5-dH4PteAPBA,N Alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-DL-2-amino-4-phosphobutanoic acid; 5-dH4PteOro,N Alpha-(5-dideaza-5,6,7,8-tetrahydropteroyl)-L-ornithine; ICI-198,583,2-desamino-2-methyl-N10-propargyl-5,8-dideazafolate; 4-H-ICI-198,583,4-deoxy-ICI-198,583:4-OCH3-ICI-198,583,4-methoxy-ICI-198,583 Glu-to-Val-ICI-198,583;Valine-ICI-198,583;Glu-to-Sub-ICI-198,583,2-aminosuberic acid-ICI-198,583;7-CH3-ICI-198,583,7-methyl-ICI-198,583;ZD1694,N-[5(N-(3,4-dihydro-2-methyl-4-oxy Soquinazoline-6-yl-methyl)amino)2-thienyl)]-L-glutamic acid; 2-NH2-ZD1694, 2-amino-ZD1694; BW1843U89, (S)-2[5-(((1,2-dihydro-3-methyl-1-oxobenzo(f)quinazoline-9-yl)methyl)amino-)-1-oxo-2-isoindlinyl]-glutaric acid; LY231514 is a polyglutamate of a folate antagonist selected from LY231514, N-(4-(2-(2-amino-4,7-dihydro-4-oxo-3H-pyrrolo[2,3-D]pyrimidine-5-yl)ethyl)-benzoyl]-L-glutamic acid; IAHQ, 5,8-dideazaisofolate; 2-dIAHQ, 2-desamino-IAHQ; 2-CH3-dIAHQ, 2-desamino-2-methyl-IAHQ; 5-d(i)PteGlu, 5-deazaisofolate; N9-CH3-5-d(i)PteGlu, N9-methyl-5-deazaisofolate; N9-CHO-5-d(i)PteGlu, N9-formyl-5-deazaisofolate, or stereoisomers thereof; (c) The liposomes encapsulate 10 to 500,000 gamma polyglutamine oxidase folate antimetabolitic antagonist molecules; (d) The liposomes are pegylated and are formed from a component comprising cholesterol, at least one neutral lipid, and at least one anionic lipid; (e) The liposomes are neutral, anionic, or cationic; (f) The liposomes have a diameter of 20 nm to 200 nm; Liposome composition.

2. The liposome composition according to claim 1, wherein the gamma-polyglutamine oxidized folate antimetabolitic agent comprises 4 to 6 glutamyl groups linked by gamma-carboxyl group bonds.

3. The polyglutamine-oxidizable folate antimetabolites that are not polyglutamine-oxidized are selected from the group consisting of methotrexate (MTX), pemetrexed (PMX), lometrexol (LMX), larcitrexed (RTX), pralatrexate, AG2034, GW1843, aminopterin, and LY309887; or The polyglutamine-inoxidizable folate antagonist is selected from the group consisting of trimethrexate (TMQ), pyritrexime (BW301U), talotrexin (PT523), noratexide (AG337), previtrexed (ZD9331, BGC9331), and BGC945 (ONX0801); The liposome composition according to claim 1.

4. (a) At least two of the glutamyl groups of the gamma polyglutamine oxoxide antimetabolite are L-type; (b) At least one of the glutamyl groups of the gamma polyglutamine oxoxide antimetabolite is of type D; or (c) At least two of the glutamyl groups of the gamma polyglutamine oxidized folate antimetabolite are L-type and at least one of the glutamyl groups is D-type; The liposome composition according to claim 1.

5. The liposome composition according to claim 1, wherein the gamma-polyglutamine oxidase antagonist is a gamma-tetraglutamine oxidase antagonist, a gamma-pentaglutamine oxidase antagonist, or a gamma-hexaglutamine oxidase antagonist.

6. The liposome composition according to claim 1, wherein the liposomes have a diameter in the range of 80 nm to 120 nm.

7. The liposome composition according to claim 1, wherein the liposomes are formed from liposome components comprising at least one selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), DSPE-polyethylene glycol (PEG), DSPE-PEG-maleimide, hydrogenated soybean phosphatidylcholine (HSPC), HSPC-PEG, cholesterol-PEG, cholesterol-maleimide, and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE).

8. The liposome composition according to claim 1, wherein the liposome components include hydrogenated soy phosphatidylcholine (HSPC), cholesterol, and pegylated lipids.

9. Liposomes have a zeta potential of zero or less; Liposomes have a zeta potential of 0 to -150 mV; or Liposomes have a zeta potential of -30 to -50 mV; The liposome composition according to claim 1.

10. The liposome composition according to claim 1, wherein the liposomes encapsulate 100,000 to 200,000 gamma polyglutamine oxidized folate antimetabolitic antagonist molecules.

11. The liposome composition according to claim 1, wherein the internal space of the liposome contains a gamma polyglutamic acid oxidase antagonist suspended in a HEPES buffer solution containing a cryoprotective agent or isotonic agent selected from trehalose, dextrose, mannitol, and sucrose.

12. A pharmaceutical composition comprising the liposome composition described in claim 1.

13. A liposome composition according to any one of claims 1 to 11 or a pharmaceutical composition according to claim 12 for use in the treatment of a disease, wherein the disease may be cancer, an immune system disorder or an infectious disease, and the disease may be an autoimmune disease, rheumatoid arthritis or an inflammatory condition.

14. A composition for use in the treatment of cancer, comprising the liposome composition according to any one of claims 1 to 11 or the pharmaceutical composition according to claim 12.

15. The composition according to claim 14 for treating a cancer selected from lung cancer, breast cancer, colorectal cancer, pancreatic cancer, gastric cancer, bladder cancer, head and neck cancer, ovarian cancer, and cervical cancer.

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