Alpha-polyglutamine oxoxide antimetabolites and their use

Abstract

To provide an alpha polyglutamated antifolate composition capable of overcoming pharmacological challenges associated with dose-limiting toxicity and treatment resistance related to antifolate therapy.SOLUTION: There is provided a liposomal composition comprising a liposome encapsulating an alpha polyglutamated antifolate, wherein the alpha polyglutamated antifolate comprises 2 to 15 glutamyl groups linked via alpha-carboxyl group bonds, wherein the liposome is pegylated, and comprises a targeting moiety having specific affinity for a surface antigen of a desired target cell.SELECTED DRAWING: None

Description

[Background technology]

[0001] This disclosure generally relates to alphapolyglutamine oxidase folate antimetabolites compositions comprising a delivery carrier such as liposomes containing the alphapolyglutamine oxidase folate antimetabolites composition, and to methods for manufacturing and using compositions for treating diseases including hyperproliferative diseases such as cancer, immune system disorders including inflammatory 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 movement of single carbon units involved in nucleotide biosynthesis and DNA repair, homocysteine ​​remethylation (Hcy), and methylation of DNA, proteins, and lipids. The only circulating form of folic acid in the blood is monoglutamate, and folic acid monoglutamate is the only form of folic acid that is transported across the cell membrane. Similarly, monoglutamate-type polyglutamic folic acid antimetabolites are also transported across the cell membrane. Once taken up into cells, intracellular folic acid is converted to polyglutamate by the enzyme folylpolygamma-glutamate synthase (FPGS).

[0003] Folic acid 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 in a lower pH environment than normal. RFC is the main transporter of folic acid antagonists at physiological pH and is widely expressed in normal and diseased cells. Therefore, folic acid antagonist therapy often suffers from dose-limiting toxicity, which is a major obstacle in cancer chemotherapy. Once inside the cell, folic acid antagonists are polyglutamylated by FPGS, which allows up to six glutamyl groups to be added during the binding of the L-gamma carboxyl group to the folic acid antagonist. L-gamma polyglutamylation of folic acid antagonists by FPGS serves at least two major therapeutic purposes: (1) it greatly enhances the affinity and inhibitory activity of folic acid antagonists for DHFR; and (2) it facilitates the accumulation of polyglutamylated folic acid antagonists, which, unlike folic acid antagonists (monoglutamic acids), are not easily transported out of the cell by cellular efflux pumps.

[0004] Targeting folic acid metabolism and nucleotide biosynthesis is an established therapeutic strategy for cancer, but for folic acid antagonists, clinical efficacy is limited due to the lack of tumor selectivity and the presence of new and acquired drug resistance. Folic acid 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 folic acid antagonist therapy and restricts the clinical application of folic acid antagonists.

[0005] Resistance to folic acid antagonist therapy is usually associated with one or more of the following: (a) increased activity of cellular efflux pumps, (b) decreased transport of folic acid 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 bound to folic acid and folic acid antagonists.

[0006] The problem with the long-term (>30 years) observation that higher levels of polyglutamate of various folic acid antagonists have much higher potency compared to lower levels of glutamate was that the scientific community has relied on the intracellular FPGS-mediated mechanism that converts lower levels of glutamate to their higher level forms. The present invention provides a means to directly deliver higher levels of polyglutamate forms of folic acid antagonists into cells without relying on the cell's machinery to achieve this goal.

[0007] The provided alpha-polyglutaminated folic acid antagonist composition provides a strategy to overcome the pharmacological issues related to dose-limiting toxicity and treatment resistance associated with folic acid antagonist therapy. In some embodiments, the provided method delivers an alpha-polyglutaminated form of a folic acid antagonist to cancer cells while simultaneously (1) minimizing / reducing exposure to normal tissue cells, (2) optimizing / improving the cytotoxic effect of the folic acid antagonist-based agent on cancer cells, and (3) minimizing / reducing the effect of efflux pumps and other resistance mechanisms that limit the therapeutic efficacy of the folic acid antagonist. SUMMARY OF THE INVENTION

[0008] The present disclosure generally relates to novel alpha-polyglutaminated folic acid antagonist (αPANTIFOL) compositions, as well as methods of manufacture and use of the compositions for treating diseases including hyperproliferative diseases such as cancer, immune system disorders such as inflammation and rheumatoid arthritis, cardiovascular diseases such as coronary artery disease, and infectious diseases such as HIV, malaria, and schistosomiasis.

[0009] In some embodiments, the present disclosure provides the following. [1] A composition comprising an alpha-polyglutaminated folic acid antagonist, wherein at least one glutamyl group has an alpha-carboxyl group bond. [2] A composition of item [1] wherein the folate antagonist is selected from pyritrexime, pralatrexate, AG2034, GW1843, and LY309887, or their stereoisomers; [3] A composition of item [1] wherein the folate antagonist is selected from PMX, MTX, RTX, and LMX, or their stereoisomers; [4] A composition described in item [1], 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, folate antagonist; 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 described in any one of items [1] to [5], wherein the composition is as follows: (a) Each glutamyl group of a polyglutamic folate antagonist, other than the glutamyl group of the folate antagonist, has an alpha-carboxyl group bond; or (b) Two or more glutamyl groups of the polyglutamine oxoxide antimetabolites have gammacarboxyl group bonds; [7] A composition described in any one of items [1] to [5], wherein the composition is as follows: (a) Each glutamyl group other than the C-terminal glutamyl group and the glutamyl group of the folate antagonist has an alpha-carboxyl group bond; or (b) Each glutamyl group other than the C-terminal glutamyl group(s) has an alpha-carboxyl group bond; [8] A composition according to any one of items [1] to [7], wherein the alpha-polyglutamine oxoxide antimetabolitic agent is as follows: (a) Containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups; (b) It is an alphapentaglutamine oxoxide antimetabolitic antagonist; or (c) It is an alpha-hexaglutamine oxoxide antimetabolite; [9] A composition according to any one of items [1] to [8], wherein the alpha-polyglutamine oxoxide antimetabolitic agent comprises 1 to 10 glutamyl groups having an alpha-carboxyl group bond;

[10] A composition described in any one of items [1] to [9], wherein the composition is as follows: (a) At least two glutamyl groups of the alpha-polyglutamine oxoxide antimetabolites are in the L-form; (b) Each glutamyl group in the alpha-polyglutamine oxoxide antimetabolites is in the L form; (c) At least one glutamyl group of the alpha-polyglutamine oxoxide antimetabolites is of the D type; (d) The glutamyl groups of each alpha-polyglutamic folate antagonist, other than the glutamyl group of the folate antagonist, are of type D; or (e) At least two glutamyl groups of the alpha-polyglutamine oxoxide antimetabolites are L-type and at least one glutamyl group is D-type;

[11] A composition according to any one of items [1] to

[10] , wherein the polyglutamate is linear;

[12] A composition according to any one of items [1] to

[10] , wherein the polyglutamate is a branched chain;

[13] A liposome composition containing an alpha-polyglutamine oxoxide antimetabolitic agent as described in any one of items [1] to

[12] (Lp-αPANTIFOL);

[14] Lp-αPANTIFOL composition of item

[13] , wherein the alpha-polyglutamine oxoxide antimetabolitic antagonist is selected from the following: (a) AG2034, pyritrexime, pralatrexate, GW1843, folate antagonists, and LY309887; or (b) PMX, MTX, RTX, and LMX, or their stereoisomers;

[15] Lp-αPANTIFOL composition of item

[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, 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-(phenysulfonyl)benzyl)ethyl)amino]quinazoline; or their stereoisomers;

[16] Lp-αPANTIFOL compositions as described in item

[13] , 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;

[17] A composition of Lp-αPANTIFOL according to any one of items

[13] to

[16] , wherein the liposomes contain an alpha-polyglutamine oxoxide antimetabolitic agent having 4, 5, 6, 2 to 10, 4 to 6, or more than 5 glutamyl groups;

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

[13] to

[17] , wherein the liposome contains an alpha-tetraglutamine oxoxide antimetabolitic agent;

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

[13] to

[17] , wherein the liposome comprises an alpha-pentaglutamine oxoxide antimetabolitic agent;

[20] Lp-αPANTIFOL composition according to any one of items

[13] to

[17] , wherein the liposome comprises an alpha-hexaglutamine oxoxide antimetabolitic agent;

[21] Lp-αPANTIFOL compositions according to any one of items

[13] to

[20] , wherein the polyglutamate is linear or branched;

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

[13] to

[21] , which: (a) Each glutamyl group other than the glutamyl group of the folate antagonist has an alpha-carboxyl group bond; or (b) A composition having two or more glutamyl groups bonded to a gammacarboxyl group;

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

[13] to

[21] , which: (a) Each glutamyl group other than the C-terminal glutamyl group and the glutamyl group of the folate antagonist has an alpha-carboxyl group bond; or (b) A composition in which each glutamyl group other than the C-terminal glutamyl group(s) has an alpha-carboxyl group bond;

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

[13] to

[23] , which: (a) At least two glutamyl groups of the alpha-polyglutamine oxoxide antimetabolites are in the L-form; (b) Each glutamyl group in the alpha-polyglutamine oxoxide antimetabolites is in the L form; (c) At least one glutamyl group of the alpha-polyglutamine oxoxide antimetabolites is of the D type; (d) The glutamyl groups of each alpha-polyglutamic folate antagonist, other than the glutamyl group of the folate antagonist, are of type D; or (e) At least two glutamyl groups of the alpha-polyglutamine oxoxide antimetabolites are L-type and at least one glutamyl group is D-type;

[25] A composition of Lp-αPANTIFOL as described in any one of items

[13] to

[24] , wherein the liposomes are pegylated (PαLp-αPANTIFOL);

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

[13] to

[24] , wherein the liposomes are not pegylated;

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

[13] to

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

[28] Lp-αPANTIFOL compositions according to any one of items

[13] to

[27] , wherein the polyglutamate is linear or branched;

[29] A Lp-αPANTIFOL composition according to any one of items

[13] to

[28] , wherein the liposome contains at least 1% by weight of an alpha-polyglutamine oxidase antagonist, or, during the process of preparing Lp-αPANTIFOL, at least 1% of a starting material of an alpha-polyglutamine oxidase antagonist is encapsulated in the Lp-αPANTIFOL;

[30] Lp-αPANTIFOL composition according to any one of items

[13] to

[29] , wherein the liposomes have a diameter in the range of 20 nm to 500 nm or 20 nm to 200 nm;

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

[13] to

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

[32] A composition of Lp-αPANTIFOL according to any one of items

[13] to

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

[33] A composition of Lp-αPANTIFOL as described in item

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

[34] Lp-αPANTIFOL compositions according to item

[32] or

[33] , 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;

[35] Lp-αPANTIFOL compositions according to any one of items

[32] to

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

[36] A composition of Lp-αPANTIFOL according to any one of items

[32] to

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

[37] A composition of Lp-αPANTIFOL as described in item

[36] , 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;

[38] A composition of Lp-αPANTIFOL as described in item

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

[39] A composition of Lp-αPANTIFOL according to any one of items

[13] to

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

[40] A composition of Lp-αPANTIFOL according to any one of items

[13] to

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

[41] An Lp-αPANTIFOL composition according to any one of items

[13] to

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

[42] Lp-αPANTIFOL composition according to any one of items

[13] to

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

[43] A composition of Lp-αPANTIFOL according to any one of items

[13] to

[38] , wherein the liposomes are cationic;

[44] A Lp-αPANTIFOL composition according to any one of items

[13] to

[43] , wherein the liposome has an internal space containing an alpha-polyglutamine oxidase antagonist and an aqueous pharmaceutically acceptable carrier;

[45] Lp-αPANTIFOL compositions of item

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

[46] Lp-αPANTIFOL composition of item

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

[47] Lp-αPANTIFOL composition of item

[46] , comprising 5% to 20% by weight of trehalose as a pharmaceutically acceptable carrier;

[48] ​​A composition of Lp-αPANTIFOL according to any one of items

[44] to

[47] , wherein the pharmaceutically acceptable carrier comprises 1% to 15% by weight of dextrose;

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

[44] to

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

[50] Lp-αPANTIFOL compositions according to any one of items

[44] to

[49] , 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;

[51] Lp-αPANTIFOL compositions according to any one of items

[44] to

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

[52] A Lp-αPANTIFOL composition according to any one of items

[13] to

[51] , 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;

[53] A Lp-αPANTIFOL composition according to any one of items

[13] to

[52] , wherein the liposomes contain less than 500,000 or less than 200,000 alpha-polyglutamine oxidase antagonist molecules;

[54] A Lp-αPANTIFOL composition according to any one of items

[13] to

[53] , wherein the liposomes contain 10 to 100,000 or any range in between alpha-polyglutamine oxidase antagonist molecules;

[55] A composition according to any one of items

[13] to

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

[56] Lp-αPANTIFOL compositions as described in item

[55] , 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;

[57] Lp-αPANTIFOL compositions of item

[55] or

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

[58] A composition of Lp-αPANTIFOL according to any one of items

[55] to

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

[59] Lp-αPANTIFOL composition according to any one of items

[55] to

[58] , wherein the targeting portion binds to a surface antigen with an equilibrium dissociation constant (Kd) in the range of 0;5x10⁻¹⁰ to 10x10⁻⁶ as measured by BIACORE® analysis;

[60] A Lp-αPANTIFOL composition according to any one of items

[55] to

[59] , wherein the targeting moiety specifically binds to one or more folate receptors selected from folate receptor alpha (FR-α), folate receptor beta (FR-β), and folate receptor delta (FR-δ);

[61] A Lp-αPANTIFOL composition according to any one of items

[55] to

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

[62] A composition of Lp-αPANTIFOL according to any one of items

[55] to

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

[63] A Lp-αPANTIFOL composition according to any one of items

[44] 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 conjugated to the PEG or outer surface of a liposome;

[64] Lp-αPANTIFOL compositions of item

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

[65] Lp-αPANTIFOL compositions of item

[63] or

[64] , wherein the immunostimulant is at least one selected from fluorescein, fluorescein isothiocyanate (FITC), DNP, beta-glucan, beta-1,3-glucan, beta-1,6-glucan, resolvin (e.g., resolvin D, resolvin E, or T-series resolvins such as Dn-6DPA or Dn-3DPA), oxidized low-density lipoprotein (e.g., OXPAC, PGPC), and Toll-like receptor (TLR) modulators such as erythritol lipids (e.g., E5564);

[66] A composition of Lp-αPANTIFOL described in any one of items

[63] to

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

[67] A composition comprising Lp-αPANTIFOL according to any one of items

[63] to

[66] , further comprising a hapten;

[68] Lp-αPANTIFOL composition of item

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

[69] A composition of Lp-αPANTIFOL according to any one of items

[13] to

[68] , further comprising at least one cryoprotective substance selected from mannitol, trehalose, sorbitol, and sucrose;

[70] Targeted compositions comprising any one of items [1] to

[69] ;

[71] Non-targeting compositions comprising any one of items [1] to

[54] and

[64] to

[69] ;

[72] A composition of Lp-αPANTIFOL according to any one of items

[13] to

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

[73] A pharmaceutical composition comprising a liposomal alpha-polyglutamine oxidized folate antimetabolitic agent composition described in any one of items

[13] to

[72] ;

[74] A pharmaceutical composition comprising an alpha-polyglutamine oxoxide antimetabolitic agent composition described in any one of items [1] to [8];

[75] A composition described in any one of items [1] to

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

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

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

[77] A method for treating or preventing a disease of which treatment or prevention is in need, comprising the step of administering a composition of any one of items [1] to

[75] to the subject;

[78] A method for treating or preventing a disease of which treatment or prevention is in need of treatment or prevention, comprising the step of administering a liposomal alpha-polyglutamine oxidized folate antimetabolitic composition described in any one of items

[13] to

[74] to the target;

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

[74] ;

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

[13] to

[74] ;

[81] A method of item

[79] or

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

[82] A method for treating cancer, comprising the step of administering an effective amount of any one of items [1] to

[74] to a subject having or at risk of having cancer;

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

[13] to

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

[84] Methods of item

[82] or

[83] 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 methods selected from, for example, hematological malignancies, including leukemia, lymphoma and other B-cell malignancies, myeloma and other plasma cell proliferation disorders;

[85] Methods of item

[82] or

[83] wherein the cancer is a member 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, or central nervous system (CNS) lymphoma;

[86] Methods of item

[82] or

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

[87] A method of item

[82] or

[83] wherein the cancer is a sarcoma such as osteosarcoma;

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

[55] to

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

[89] Maintenance therapy comprising the step of administering an effective amount of any one of items [1] to

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

[90] Maintenance therapy comprising the step of administering an effective amount of a liposomal alpha-polyglutamine oxidized folate antimetabolitic composition described in any one of items

[13] to

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

[91] A method for treating an immune system disorder comprising the step of administering an effective amount of any one of items [1] to

[74] 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;

[92] A method for treating an immune system disorder comprising the step of administering an effective amount of a liposomal alpha-polyglutamine oxidized folate antimetabolitic composition described in any one of items [9] to

[74] to a subject having or at risk of having an immune system disorder, optionally 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 arteriovenous disease, and psoriasis;

[93] Treatment methods below: (a) A method for treating an infectious disease, comprising the step of administering an effective amount of any one of items [1] to

[74] 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 one of items [1] to

[74] 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 one of items [1] to

[74] 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 one of items [1] to

[74] 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 one of items [1] to

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

[74] to a subject having or at risk of having a skin disease, wherein the skin disease is psoriasis;

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

[13] to

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

[95] A method for delivering an alpha-polyglutamine oxidase antagonist to a tumor expressing a folate receptor on its surface, comprising the step of administering an Lp-αPANTIFOL composition described in any one of items [1] to

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

[96] A method for preparing an alphapolyglutamine oxidase antagonist composition comprising a liposome alphapolyglutamine oxidase antagonist composition described in any one of items

[13] to

[74] , comprising the steps of: forming a mixture containing liposome components and an alphapolyglutamine oxidase antagonist in solution; homogenizing the mixture in solution to form liposomes; and processing the mixture to form liposomes containing an alphapolyglutamine oxidase antagonist;

[97] A method for preparing an alphapolyglutamine oxidase antagonist composition comprising a liposome alphapolyglutamine oxidase antagonist composition described in any one of items

[13] to

[74] , comprising the steps of: forming a mixture in solution containing a liposome component and an alphapolyglutamine oxidase antagonist; and processing the mixture to form liposomes containing the alphapolyglutamine oxidase antagonist;

[98] A method of item

[97] wherein the step of processing the mixture comprises the step of homogenizing the mixture in solution to form liposomes;

[99] A method for preparing a composition according to any one of items

[55] to

[74] , comprising the steps of: forming a mixture in solution containing a liposome component and an alpha-polyglutamate oxidase antagonist; homogenizing the mixture in solution to form liposomes; processing the mixture to form liposomes that encapsulate and / or contain the alpha-polyglutamate oxidase antagonist; and imparting a targeting moiety to 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);

[0100] A method for preparing a composition according to any one of items

[55] to

[74] , comprising the steps of: forming a mixture in solution containing a liposome component and an alpha-polyglutamate oxidase antagonist; processing the mixture to form liposomes that encapsulate and / or contain the alpha-polyglutamate oxidase antagonist; and imparting a targeting moiety to 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);

[0101] A method according to item

[0100] , wherein the processing step includes homogenizing a mixture in solution to form liposomes;

[0102] A method according to any one of items

[99] to

[0101] , 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;

[0103] A method according to any one of items

[99] to

[0102] , 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

[0104] A method according to any one of items

[96] to

[0103] , wherein at least 1% of an alpha-polyglutamine oxoxide antimetabolitic antagonist starting material is encapsulated or enclosed in Lp-αPANTIFOL.

[0010] In some embodiments, the Disclosure provides alpha-polyglutamic acid folate antimetabolites (αPANTIFOL) compositions in which at least one glutamyl residue of the alpha-polyglutamic acid folate antimetabolites is bonded by an alpha-carboxyl group. In some embodiments, αPANTIFOL comprises 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 antimetabolites). In some embodiments, the alpha-polyglutamic acid folate antimetabolites are selected from (a) AG2034, pyritrexime, pralatrexate, GW1843, folate antimetabolites, and LY309887; or (b) PMX, MTX, RTX, and LMX, or their stereoisomers. In some embodiments, the alpha-polyglutamic acid 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 alpha-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, αPANTIFOL comprises two or more L-type glutamyl groups. In other embodiments, αPANTIFOL comprises a D-type glutamyl group. In further embodiments, αPANTIFOL comprises a D-type glutamyl group and two or more L-type glutamyl groups. In further embodiments, αPANTIFOL comprises two or more glutamyl groups having a gamma bond.

[0011] In one embodiment, the αPANTIFOL composition comprises a chain of three glutamyl groups bonded to a glutamyl group in the folate antagonist (i.e., a tetraglutamate folate antagonist). In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [2] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [3] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [4] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [5] of the Summary of Invention section. In some embodiments, αPANTIFOL 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. In further embodiments, the alpha-tetraglutamine oxidase antagonist contains two or more glutamyl groups having a gamma bond.

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

[0013] In one embodiment, the αPANTIFOL composition comprises a chain of five glutamyl groups bonded to a glutamyl group in a folate antagonist (i.e., a hexaglutamic folate antagonist). In some embodiments, αPANTIFOL is a polyglutamate of a folate antagonist as described in item [2] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamate of a folate antagonist as described in item [3] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamate of a folate antagonist as described in item [4] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamic folate antagonist as described in item [5] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamic folate antagonist as described in the Summary of Invention section. In some embodiments, the hexaglutamine oxidase antagonist comprises two or more L-type glutamyl groups. In other 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 pentagulutamine 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. In further embodiments, the alpha-hexaglutamine oxidase antagonist comprises two or more glutamyl groups having a gamma bond.

[0014] In further embodiments, the disclosure provides compositions comprising delivery carriers such as liposomes filled (i.e., encapsulated) and / or otherwise conjugated thereto with alpha-polyglutamate folate antagonists, and methods for preparing αPANTIFOL-filled / conjugated delivery carrier compositions (DV-αPANTIFOL) and methods for using them to deliver alpha-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 αPANTIFOL-filled / conjugated 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 alpha-polyglutamic acid folate antagonist in DV-αPANTIFOL comprises 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 comprises the polyglutamic acid folate antagonist described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, the delivery carrier comprises the polyglutamic acid folate antagonist described in the Summary of the Invention section.

[0015] In further embodiments, the disclosure provides compositions (Lp-αPANTIFOL) comprising liposomes encapsulated (filled) with an alpha-polyglutamate folate antagonist. In some embodiments, the alpha-polyglutamate folate antagonist in Lp-αPANTIFOL 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 alpha-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 LMX, or their stereoisomers. In some embodiments, the liposome-encapsulated alpha-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 alpha-polyglutamic acid folate antagonist encapsulated by liposomes 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. In some embodiments, the alpha-polyglutamic acid folate antagonist in Lp-αPANTIFOL contains two or more L-type glutamyl groups. In other embodiments, the alpha-polyglutamic acid folate antagonist in Lp-αPANTIFOL contains a D-type glutamyl group. In further embodiments, the alpha-polyglutamic acid folate antagonist in Lp-αPANTIFOL contains a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the alpha-polyglutamic acid folate antagonist in Lp-αPANTIFOL contains two or more glutamyl groups having gamma bonds. In further embodiments, the alpha-polyglutamic acid folate antagonist in Lp-αPANTIFOL contains one or more glutamyl groups having both alpha and gamma bonds. In some embodiments, the alpha-polyglutamic acid folate antagonist in Lp-αPANTIFOL contains 2 to 10 or any range in between, glutamyl groups having both alpha and gamma bonds. In some embodiments, the polyglutamate chain of the alpha-polyglutamic acid folate antagonist is linear. In some embodiments, the polyglutamate chain of the alpha-polyglutamine oxoxide antimetabolites is branched.

[0016] In one embodiment, the Lp-αPANTIFOL composition comprises an alpha-polyglutamic folate antagonist containing a chain of three glutamyl groups bonded to a glutamyl group in the folate antagonist (i.e., a tetraglutamic folate antagonist). In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [2] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [3] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [4] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamic folate antagonist described in item [5] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamic 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. In further embodiments, the tetraglutamine oxidase antagonist comprises two or more glutamyl groups having a gamma bond. In some embodiments, the polyglutamate chain of the alphapolyglutamine oxidase antagonist is linear. In some embodiments, the polyglutamate chain of the alphapolyglutamine oxidase antagonist is branched.

[0017] In one embodiment, the Lp-αPANTIFOL composition comprises an alpha-polyglutamic folate antagonist containing a chain of four glutamyl groups bonded to a glutamyl group in the folate antagonist (i.e., a pentaglutamic folate antagonist). In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [2] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [3] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [4] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamic folate antagonist described in item [5] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamic 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 other embodiments, the pentaglobulin 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. In further embodiments, the pentaglobulin oxidase antagonist comprises two or more glutamyl groups having a gamma bond. In some embodiments, the polyglutamate chain of the alpha-polyglutamulin oxidase antagonist is linear. In some embodiments, the polyglutamate chain of the alpha-polyglutamulin oxidase antagonist is branched.

[0018] In one embodiment, the Lp-αPANTIFOL composition comprises an alpha-polyglutamic folate antagonist containing a chain of five glutamyl groups bonded to a glutamyl group in the folate antagonist (i.e., a hexaglutamic folate antagonist). In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [2] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [3] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [4] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamic folate antagonist described in item [5] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamic folate antagonist described in the Summary of Invention section. In some embodiments, the hexaglutamine oxidase antagonist contains two or more L-type glutamyl groups. In other embodiments, the hexaglutamine oxidase antagonist contains a D-type glutamyl group. In some embodiments, the hexaglutamine oxidase antagonist contains two or more D-type glutamyl groups. In further embodiments, the hexaglutamine oxidase antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the pentagramamine oxidase antagonist contains one, two, three, four, or five D-type glutamyl groups and five, four, three, two, or one L-type glutamyl group, respectively. In further embodiments, the hexaglutamine oxidase antagonist contains two or more glutamyl groups having a gamma bond. In some embodiments, the polyglutamate chain of the alpha-polyglutamine oxidase antagonist is linear. In some embodiments, the polyglutamate chain of the alpha-polyglutamic folate antimetabolites is branched.

[0019] In some embodiments, the Lp-αPANTIFOL composition is cationic. In some embodiments, the Lp-αPANTIFOL liposomes are cationic and have a diameter in the range of 20 nm to 500 nm, 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-αPANTIFOL liposomes are cationic and the composition has a diameter in the range of 80 nm to 120 nm, or any range in between. In some embodiments, the cationic Lp-αPANTIFOL composition contains at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% w / w or more than 75% w / w of alpha-polyglutamic oxidase folate antimetabolites. In some embodiments, during the preparation of Lp-αPANTIFOL, at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or more than 75% of the starting material of the alpha-polyglutamate oxidase antagonist is encapsulated in cationic Lp-αPANTIFOL. In further embodiments, the alpha-polyglutamate oxidase antagonist encapsulated by liposomes is present in HEPES buffer within the liposomes.

[0020] In other embodiments, the Lp-αPANTIFOL composition is anionic or neutral. In some embodiments, the Lp-αPANTIFOL composition is cationic. In some embodiments, the Lp-αPANTIFOL liposomes are anionic or neutral and have a diameter in the range of 20 nm to 500 nm, 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-αPANTIFOL 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-αPANTIFOL liposomes are anionic and have a diameter in the range of 20 nm to 500 nm, 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-αPANTIFOL 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-αPANTIFOL liposomes are neutral, and have a diameter in the range of 20 nm to 500 nm, 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-αPANTIFOL 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-αPANTIFOL composition contains at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% w / w or more than 75% w / w of alpha-polyglutamic acid folate antimetabolites. In some embodiments, during the preparation of Lp-αPANTIFOL, at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% or more of alpha-polyglutamic acid folate antimetabolites are encapsulated in the anionic or neutral Lp-αPANTIFOL.In some embodiments, the anionic or neutral Lp-αPANTIFOL composition contains at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% w / w or more than 75% w / w of alpha-tetraglutamine oxidase antagonist. In some embodiments, the anionic or neutral Lp-αPANTIFOL composition contains at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% w / w or more than 75% w / w of alpha-pentaglutamine oxidase antagonist. In some embodiments, the anionic or neutral Lp-αPANTIFOL composition contains at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% w / w or more than 75% w / w of the alpha-hexaglutamine oxidase antagonist. In further embodiments, the alpha-polyglutamine oxidase antagonist encapsulated by liposomes is present in HEPES buffer within the liposomes.

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

[0022] In some embodiments, the liposomal alpha-polyglutamine oxidase antagonist composition is untargeted (NTLp-αPANTIFOL). That is, the NTLp-αPANTIFOL composition does not have specific affinity for epitopes expressed on the surface of target cells of interest (e.g., epitopes on surface antigens). In some embodiments, the NTLp-αPANTIFOL composition does not contain a targeting moiety. In further embodiments, the untargeted liposomal alpha-polyglutamine oxidase antagonist composition is pegylated (NTPLp-αPANTIFOL).

[0023] In other embodiments, the liposomal alpha-polyglutamine oxidized folate antimetabolitician composition is targeted (TLp-αPANTIFOL). That is, the TLp-αPANTIFOL composition includes a targeting moiety having specific affinity for an epitope (surface antigen) on the target cell of interest. In some embodiments, the targeting moiety of TLp-αPANTIFOL or TPLp-αPANTIFOL is not covalently bound to the liposome. In other embodiments, the targeting moiety of TLp-αPANTIFOL or TPLp-αPANTIFOL is bound to one or both of the PEG and outer surface of the liposome. In some embodiments, the targeting moiety of TLp-αPANTIFOL or TPLp-αPANTIFOL is covalently bound to the liposome. The functions of the targeting moiety of TLp-αPANTIFOL and / or TPLp-αPANTIFOL 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 (αPANTIFOL) 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 alpha-polyglutamate folate antimetabolites (TLp-αPANTIFOL and TPLp-αPANTIFOL) provide further improvements to the efficacy and safety profiles of folate antimetabolites by specifically delivering alpha-polyglutamate (e.g., tetraglutamate, pentaglutamate, and hexaglutamate) folate antimetabolites to target cells such as cancer cells. In further embodiments, the targeted liposomal alpha-polyglutamate folate antimetabolites are pegylated (TPLp-αPANTIFOL). The functions of the targeting portion of the TLp-αPANTIFOL and / or TPLp-αPANTIFOL composition include, but are not limited to, targeting liposomes to target cells of interest in vivo or in vitro; interacting with surface antigens with specific affinity for the targeting portion; and delivering the liposomal payload (αPANTIFOL) to the cells.

[0025] Suitable 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. In some embodiments, the targeting moiety is an antibody or an antigen-binding antibody fragment. In further embodiments, the targeting moiety comprises one or more of antibodies, humanized antibodies, antigen-binding fragments of antibodies, single-chain antibodies, single-domain antibodies, bispecific antibodies, synthetic antibodies, pegylated antibodies, and multimeric antibodies. In some embodiments, the targeting moiety of TLp-αPANTIFOL or TPLp-αPANTIFOL 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 on or difficult to access on non-tumor cells. In some embodiments, the targeting portion is measured by BIACORE® analysis and has an affinity of 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-αPANTIFOL or TPLp-αPANTIFOL 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 is 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 FR-α and FR-β.

[0027] In further embodiments, the αPANTIFOL composition comprises one or more immunostimulants, detectable markers, and maleimides, positioned on at least one of the PEGs or outer surfaces of the liposome. In some embodiments, the liposomal αPANTIFOL composition (e.g., Lp-αPANTIFOL, PLp-αPANTIFOL, NTLp-αPANTIFOL, NTPLp-αPANTIFOL, TLp-αPANTIFOL, or TPLp-αPANTIFOL) is cationic. In other embodiments, the liposomal αPANTIFOL composition (e.g., Lp-αPANTIFOL, PLp-αPANTIFOL, NTLp-αPANTIFOL, NTPLp-αPANTIFOL, TLp-αPANTIFOL, or TPLp-αPANTIFOL) is anionic or neutral. In further embodiments, the liposomes of the liposome-αPANTIFOL composition (e.g., Lp-αPANTIFOL, PLp-αPANTIFOL, NTLp-αPANTIFOL, NTPLp-αPANTIFOL, TLp-αPANTIFOL, or TPLp-αPANTIFOL) 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-αPANTIFOL composition have a diameter in the range of 80 nm to 120 nm, or any range in between. In some embodiments, the liposome-αPANTIFOL composition is pegylated (e.g., PLp-αPANTIFOL, NTPLp-αPANTIFOL, or TPLp-αPANTIFOL). In some embodiments, the liposome-αPANTIFOL composition includes a targeting moiety (e.g., TLp-αPANTIFOL or TPLp-αPANTIFOL). In further embodiments, the liposomal αPANTIFOL composition is pegylated and targeted (e.g., TPLp-αPANTIFOL). In some embodiments, the liposomal αPANTIFOL composition comprises an alpha-polyglutamic acid folate antagonist containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomal αPANTIFOL composition comprises an alpha-tetraglutamic acid folate antagonist.In some embodiments, the liposomal αPANTIFOL composition comprises an alpha-pentaglutamine oxidase folate antagonist. In other embodiments, the liposomal αPANTIFOL composition comprises an alpha-hexaglutamine oxidase folate antagonist. In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [2] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [3] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [4] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamate of the folate antagonist described in item [5] of the Summary of Invention section. In some embodiments, αPANTIFOL is a polyglutamamine oxidase folate antagonist described in the Summary of Invention section.

[0028] In some embodiments, the liposome composition comprises an alpha-polyglutamic acid folate antagonist containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups and an alpha-polyglutamic acid folate antagonist in at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% w / w, or more than 75% w / w. In some embodiments, the Lp-αPANTIFOL composition comprises an alpha-polyglutamic acid folate antagonist containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups and an alpha-polyglutamic acid folate antagonist in 1% to 98.5% w / w. In some embodiments, the liposomes contain an alpha-polyglutamic acid folate antagonist comprising 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups, and during the preparation of Lp-αPANTIFOL, at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or more than 75% of the alpha-polyglutamic acid folate antagonist starting material is encapsulated in Lp-αPANTIFOL. In some embodiments, the liposome composition comprises an alpha-polyglutamic acid folate antagonist as described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, the liposome composition comprises a liposome as described in any one of items

[13] to

[72] in the Summary of the Invention section. In some embodiments, the composition comprises an alpha-polyglutamine oxidase antagonist described in the summary section of the invention.

[0029] In further embodiments, the liposomal αPANTIFOL composition (i.e., Lp-αPANTIFOL such as PLp-αPANTIFOL, NTLp-αPANTIFOL, NTPLp-αPANTIFOL, TLp-αPANTIFOL, or TPLp-αPANTIFOL) contains at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of liposomal encapsulated alpha-polyglutamate oxidase antagonist. In some embodiments, the liposomal αPANTIFOL composition contains 1% to 98.5% of liposomal encapsulated alpha-polyglutamate oxidase antagonist. In further embodiments, the liposomal αPANTIFOL 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 alpha-polyglutamic acid folate antagonist containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomal αPANTIFOL composition comprises 1% to 98.5% of a liposomal encapsulated alpha-polyglutamic acid folate antagonist containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomal αPANTIFOL composition comprises at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of the liposomally encapsulated alpha-tetraglutamine oxidase antagonist. In some embodiments, the liposomal composition comprises the alpha-polyglutamine oxidase antagonist described in any one of items [1] to

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

[13] to

[72] in the Summary of the Invention section. In some embodiments, the liposomal composition comprises the alpha-polyglutamine oxidase antagonist described in the Summary of the Invention section.

[0030] In some embodiments, the liposome composition comprises an alpha-tetraglutamine oxidase antagonist and at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% w / w or more than 75% w / w of the alpha-tetraglutamine oxidase antagonist. In some embodiments, the Lp-αPANTIFOL composition comprises an alpha-tetraglutamine oxidase antagonist and 1% to 98.5% w / w of the alpha-tetraglutamine oxidase antagonist. In some embodiments, the liposomes contain an alpha-tetraglutamine oxidase antagonist, and during the preparation of Lp-αPANTIFOL, at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or more than 75% of the alpha-tetraglutamine oxidase antagonist starting material is encapsulated in Lp-αPANTIFOL. In some embodiments, the liposome composition contains an alpha-polyglutamine oxidase antagonist as described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, the liposome composition contains a liposome as described in any one of items

[13] to

[72] in the Summary of the Invention section. In some embodiments, the liposome composition contains an alpha-polyglutamine oxidase antagonist as described in the Summary of the Invention section.

[0031] In some embodiments, the liposome composition comprises an alpha-pentaglutamine oxidase antagonist, comprising at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% w / w or more than 75% w / w of the alpha-pentaglutamine oxidase antagonist. In some embodiments, the Lp-αPANTIFOL composition comprises an alpha-pentaglutamine oxidase antagonist and 1% to 98.5% w / w of the alpha-pentaglutamine oxidase antagonist. In some embodiments, the liposomes contain an alpha-pentaglutamine oxidase antagonist, and during the Lp-αPANTIFOL preparation process, at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or more than 75% of the alpha-pentaglutamine oxidase antagonist starting material is encapsulated in the Lp-αPANTIFOL. In some embodiments, the liposome composition comprises an alpha-hexaglutamine oxidase antagonist, comprising at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% w / w or more than 75% w / w of the alpha-hexaglutamine oxidase antagonist. In some embodiments, the Lp-αPANTIFOL composition comprises an alpha-hexaglutamine oxidase antagonist and 1% to 98.5% w / w of the alpha-hexaglutamine oxidase antagonist. In some embodiments, the liposomes contain an alpha-hexaglutamine oxidase antagonist, and during the Lp-αPANTIFOL preparation process, at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or more than 75% of the alpha-pentaglutamine oxidase antagonist starting material is encapsulated in the Lp-αPANTIFOL. In some embodiments, the liposomal αPANTIFOL composition contains 1% to 98.5% of the liposomally encapsulated alpha-pentaglutamine oxidase antagonist.In some embodiments, the liposome composition comprises an alpha-polyglutamine oxidase antagonist described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, the liposome composition comprises a liposome described in any one of items

[13] to

[72] in the Summary of the Invention section. In some embodiments, the liposome composition comprises an alpha-polyglutamine oxidase antagonist described in the Summary of the Invention section.

[0032] In some embodiments, the liposomal αPANTIFOL composition contains 1% to 98.5% of liposomal encapsulated alpha-tetraglutamine oxidase antagonist. In some embodiments, the liposomal αPANTIFOL composition contains at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of liposomal encapsulated alpha-pentaglutamine oxidase antagonist. In some embodiments, the liposomal αPANTIFOL composition contains 1% to 98.5% of liposomal encapsulated alpha-pentaglutamine oxidase antagonist. In some embodiments, the liposomal αPANTIFOL composition comprises at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75% of the liposomally encapsulated alpha-hexaglutamine oxidase antagonist. In some embodiments, the liposomal composition comprises the alpha-polyglutamine oxidase antagonist described in any one of items [1] to

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

[13] to

[72] in the Summary of the Invention section. In some embodiments, the liposomal composition comprises the alpha-polyglutamine oxidase antagonist described in the Summary of the Invention section.

[0033] Liposome compositions comprising αPANTIFOL-encapsulated liposomes are also provided. In some embodiments, the liposome composition comprises a pegylated αPANTIFOL composition. In some embodiments, the liposome composition comprises an αPANTIFOL composition linked to or otherwise conjugated to a targeting moiety. In further embodiments, the liposome composition comprises a pegylated αPANTIFOL composition linked to or otherwise conjugated to a targeting moiety. In some embodiments, the liposome composition comprises αPANTIFOL comprising 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposome composition comprises an alpha-tetraglutamine oxidase folate antagonist. In some embodiments, the liposome composition comprises an alpha-pentaglutamine oxidase folate antagonist. In other embodiments, the liposome composition comprises an alpha-hexaglutamine oxidase folate antagonist. In some embodiments, the liposome composition comprises an alpha-polyglutamine oxidase antagonist described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, the liposome composition comprises a liposome described in any one of items

[13] to

[72] in the Summary of the Invention section. In some embodiments, the liposome composition comprises an alpha-polyglutamine oxidase antagonist described in the Summary of the Invention section.

[0034] In some embodiments, the liposome composition comprises liposomal αPANTIFOL (e.g., Lp-αPANTIFOL, PLp-αPANTIFOL, NTLp-αPANTIFOL, NTPLp-αPANTIFOL, TLp-αPANTIFOL, and TPLp-αPANTIFOL). In some embodiments, the liposomal αPANTIFOL is pegylated (e.g., NTPLp-αPANTIFOL and TPLp-αPANTIFOL). In some embodiments, the pharmaceutical composition comprises αPANTIFOL containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the pharmaceutical composition comprises an alpha-tetraglutamine oxidase antagonist. In some embodiments, the pharmaceutical composition comprises an alpha-pentaglutamine oxidase antagonist. In other embodiments, the pharmaceutical composition comprises an alpha-hexaglutamine oxidase antagonist. In some embodiments, the liposome composition comprises an alpha-polyglutamine oxidase antagonist described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, the liposome composition comprises a liposome described in any one of items

[13] to

[72] in the Summary of the Invention section. In some embodiments, the liposome composition comprises an alpha-polyglutamine oxidase antagonist described in the Summary of the Invention section. In some embodiments, the liposome αPANTIFOL 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., TLp-αPANTIFOL or TPLp-αPANTIFOL). In further embodiments, the liposome composition comprises a pegylated liposome αPANTIFOL further comprising 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-αPANTIFOL). In some embodiments, the liposome composition comprises a cationic liposome αPANTIFOL. In other embodiments, the liposome composition comprises liposome α-PANTIFOL, which is anionic or neutral.In further embodiments, the liposome composition comprises liposome αPANTIFOL having a diameter in the range of 20 nm to 500 nm, 20 nm to 200 nm, or any range in between. In further embodiments, the liposome αPANTIFOL has a diameter in the range of 80 nm to 120 nm, or any range in between.

[0035] Pharmaceutical compositions are also provided that include an alpha-polyglutamine oxidase antagonist (αPANTIFOL) containing a delivery carrier such as liposomal αPANTIFOL. In some embodiments, the pharmaceutical composition includes a pegylated αPANTIFOL composition. In some embodiments, the pharmaceutical composition includes an αPANTIFOL composition linked to or otherwise conjugated to a targeting moiety. In further embodiments, the pharmaceutical composition includes a pegylated αPANTIFOL composition linked to or otherwise conjugated to a targeting moiety. In some embodiments, the pharmaceutical composition includes αPANTIFOL containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the pharmaceutical composition includes an alpha-tetraglutamine oxidase antagonist. In some embodiments, the pharmaceutical composition includes an alpha-pentaglutamine oxidase antagonist. In other embodiments, the pharmaceutical composition includes an alpha-hexaglutamine oxidase antagonist. In other embodiments, the pharmaceutical composition includes an alpha-hexaglutamine oxidase antagonist. In some embodiments, the composition comprises an alpha-polyglutamine oxidase antagonist described in any one of items [1] to

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

[13] to

[74] in the Summary of the Invention section. In some embodiments, the pharmaceutical composition comprises an alpha-polyglutamine oxidase antagonist described in the Summary of the Invention section. In some embodiments, the pharmaceutical composition comprises a polyglutamine oxidase antagonist described in the Summary of the Invention section.

[0036] In some embodiments, the pharmaceutical composition comprises liposomal αPANTIFOL (e.g., Lp-αPANTIFOL, PLp-αPANTIFOL, NTLp-αPANTIFOL, NTPLp-αPANTIFOL, TLp-αPANTIFOL, and TPLp-αPANTIFOL). In some embodiments, the liposomal αPANTIFOL composition is pegylated (e.g., NTPLp-αPANTIFOL and TPLp-αPANTIFOL). In some embodiments, the liposomal αPANTIFOL 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., TLp-αPANTIFOL or TPLp-αPANTIFOL). In further embodiments, the pharmaceutical composition comprises a pegylated liposomal αPANTIFOL composition, further comprising 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-αPANTIFOL). In some embodiments, the pharmaceutical composition comprises a cationic liposomal αPANTIFOL. In other embodiments, the pharmaceutical composition comprises an anionic or neutral liposomal αPANTIFOL. In further embodiments, the pharmaceutical composition comprises a liposomal αPANTIFOL having a diameter of 20 nm to 500 nm or 20 nm to 500 nm, or any range in between. In further embodiments, the liposomal αPANTIFOL 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 an αPANTIFOL containing 4, 5, 6, 2 to 10, 4 to 6, or more than 5 glutamyl groups. In some embodiments, the pharmaceutical composition comprises an alpha-tetraglutamine oxidase folate antagonist. In some embodiments, the pharmaceutical composition comprises an alpha-pentaglutamine oxidase folate antagonist. In some embodiments, the pharmaceutical composition comprises an alpha-polyglutamine oxidase folate antagonist as described in any one of items [1] to

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

[13] to

[74] in the summary section of the invention.In some embodiments, the pharmaceutical composition comprises an alpha-polyglutamine oxidase antagonist described in the summary section of the invention.

[0037] 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 an alpha-polyglutamine oxidase antagonist (αPANTIFOL) 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, αPANTIFOL comprises 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the αPANTIFOL composition comprises an alpha-tetraglutamine oxidase antagonist. In some embodiments, the αPANTIFOL composition comprises an alpha-pentaglutamine oxidase antagonist. In other embodiments, the αPANTIFOL composition comprises an alpha-hexaglutamine oxidase antagonist. In some embodiments, the αPANTIFOL composition comprises an alpha-polyglutamine oxidase antagonist described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, the composition comprises a liposome composition described in any one of items

[13] to

[72] in the Summary of the Invention section. In some embodiments, the composition comprises an alpha-polyglutamine oxidase antagonist described in the Summary of the Invention section.

[0038] 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 an alpha-polyglutamine oxidase antagonist (αPANTIFOL) 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, αPANTIFOL contains 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the αPANTIFOL composition contains an alpha-tetraglutamine oxidase antagonist. In some embodiments, the αPANTIFOL composition contains an alpha-pentaglutamine oxidase antagonist. In other embodiments, the αPANTIFOL composition contains an alpha-hexaglutamine oxidase antagonist. In some embodiments, the liposome comprises an alpha-polyglutamine oxidase antagonist described in any one of items [1] to

[12] of the Summary of the Invention section. In some embodiments, the liposome is a liposome described in any one of items

[13] to

[72] of the Summary of the Invention section. In some embodiments, the liposome comprises an alpha-polyglutamine oxidase antagonist described in the Summary of the Invention section.

[0039] In further embodiments, the disclosure provides a method for killing cells, the method comprising the step of contacting cells with a composition comprising an alpha-polyglutamic oxoxide antimetabolitic (α-PANTIFOL) 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, cancer 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) lymphoma. In some embodiments, the method is carried out in vivo. In other embodiments, the method is carried out in vitro. In some embodiments, αPANTIFOL contains 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the αPANTIFOL composition contains an alpha-tetraglutamine oxidase antagonist. In some embodiments, the αPANTIFOL composition contains an alpha-pentaglutamine oxidase antagonist. In other embodiments, the αPANTIFOL composition comprises an alpha-hexaglutamine oxidase antagonist. In some embodiments, αPANTIFOL is a polyglutamine oxidase antagonist described in any one of items [1] to

[12] of the Summary of the Invention section.In some embodiments, αPANTIFOL is a polyglutamic oxidase antagonist described in the Summary of the Invention section. In some embodiments, the αPANTIFOL composition comprises liposomes described in any one of items

[13] to

[72] of the Summary of the Invention section.

[0040] In further embodiments, the present disclosure provides a method for killing cells, the method comprising the step of contacting cells with liposomes containing an alpha-polyglutamic acid oxidase antagonist (e.g., Lp-αPANTIFOL such as PLp-αPANTIFOL, NTLp-αPANTIFOL, NTPLp-αPANTIFOL, TLp-αPANTIFOL, or TPLp-αPANTIFOL). 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 hyperproliferating 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) lymphoma. 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 αPANTIFOL containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomes contain an alpha-tetraglutamine oxidase antagonist. In some embodiments, the liposomes contain an alpha-pentaglutamine oxidase antagonist. In other embodiments, the liposomes contain an alpha-hexaglutamine oxidase folate antimetabolite.In some embodiments, the liposome comprises a polyglutamic acid folate antagonist described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, the liposome comprises 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 one of items

[13] to

[72] in the Summary of the Invention section.

[0041] In further embodiments, the present disclosure provides a method for treating cancer, comprising the step of administering an effective amount of a delivery carrier (e.g., an antibody-immune complex or liposome) containing an alpha-polyglutamine oxidase antimetabolitic antagonist 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-αPANTIFOL such as PLp-αPANTIFOL, NTLp-αPANTIFOL, NTPLp-αPANTIFOL, TLp-αPANTIFOL, or TPLp-αPANTIFOL). 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 αPANTIFOL containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the delivered delivery carrier includes an alpha-tetraglutamine oxidase antagonist. In some embodiments, the delivered delivery carrier includes an alpha-pentaglutamine oxidase antagonist. In other embodiments, the delivered delivery carrier includes an alpha-hexaglutamine oxidase antagonist. In some embodiments, the administered delivery carrier comprises an L-alphapolyglutamic acid folate antagonist. In some embodiments, the administered delivery carrier comprises two, three, four, five, or more than five L-alphaglutamyl groups. In some embodiments, the administered delivery carrier comprises a D-alphapolyglutamic acid folate antagonist. In some embodiments, the administered delivery carrier comprises two, three, four, five, or more than five D-alphaglutamyl groups. In some embodiments, the administered delivery carrier comprises L and D-alphapolyglutamic acid folate antagonists. In some embodiments, the administered delivery carrier comprises two, three, four, five, or more L-alphaglutamyl groups and two, three, four, five, or more than five D-alphaglutamyl 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 (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 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) cancer.

[0042] In some embodiments, the cancer cells to be contacted are primary cells or cells derived from cell lines obtained from / originating 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 / originating 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 / originating colorectal cancer. In some embodiments, the cancer cells to be contacted are primary cells or cells derived from cell lines obtained from / originating ovarian cancer. In some embodiments, the cancer cells to be contacted are primary cells or cells derived from cell lines obtained from / originating endometrial cancer. In some embodiments, the cancer cells to be contacted are primary cells or cells derived from cell lines obtained from / originating pancreatic cancer. In some embodiments, the cancer cells to be contacted are primary cells or cells derived from cell lines obtained from / originating liver cancer. In some embodiments, the cancer cells to be contacted are primary cells or cells derived from cell lines obtained from / derived from head and neck cancer. In some embodiments, the cancer cells to be contacted are primary cells or cells derived from cell lines obtained from / derived from osteosarcoma. In some embodiments, the delivery carrier to be administered comprises a polyglutamic acid oxidase antagonist as described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, the delivery carrier comprises a polyglutamic acid oxidase antagonist as described in the Summary of the Invention section. In some embodiments, the liposome composition comprises liposomes as described in any one of items

[13] to

[72] in the Summary of the Invention section.

[0043] In further embodiments, the present disclosure provides a method for treating cancer, the method comprising the step of administering an effective amount of liposomes containing an alpha-polyglutamic acid oxidase antagonist (e.g., Lp-αPANTIFOL such as PLp-αPANTIFOL, NTLp-αPANTIFOL, NTPLp-αPANTIFOL, TLp-αPANTIFOL, or TPLp-αPANTIFOL) to a subject having or at risk of having cancer. In some embodiments, the liposomes are pegylated. In some embodiments, the liposomes are not pegylated. In further embodiments, the liposomes include a targeting moiety having specific affinity for an antigen epitope on the surface of cancer cells. In further 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, fibronectin Xtradomain 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, CD40L, 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 liposome includes a targeting moiety that specifically binds to a cell surface antigen that is determined to originate from or be expressed on a tumor of a particular target, such as a neoantigen. In some embodiments, the targeting moiety is an antibody or an antigen-binding antibody fragment. In some embodiments, the liposome includes αPANTIFOL containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposome includes an alpha-tetraglutamine oxidase antagonist. In some embodiments, the liposome includes an alpha-pentaglutamine oxidase antagonist. In other embodiments, the liposome comprises an alpha-hexaglutamate oxidase antagonist. In some embodiments, the liposome comprises a polyglutamate oxidase antagonist as described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, αPANTIFOL is a polyglutamate oxidase antagonist as described in the Summary of the Invention section. In some embodiments, the liposome composition is a liposome as described in any one of items

[13] to

[72] in the Summary of the Invention section. In some embodiments, the liposome comprises two, three, four, five, or more than five L-alphaglutamyl groups. In some embodiments, the liposome comprises a D-alphapolyglutamate oxidase antagonist. In some embodiments, the liposome comprises two, three, four, five, or more than five D-alphaglutamyl groups. In some embodiments, the liposome comprises L and D-alphapolyglutamate oxidase antagonists. In some embodiments, the liposomes contain two, three, four, five, or more than five L-alpha-glutamyl groups and two, three, four, five, or more than five D-alpha-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 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.

[0044] In a further embodiment, the present disclosure provides a method for treating cancer, the method comprising administering to a subject having or at risk of having cancer an effective amount of a liposomal composition comprising a liposome comprising an alpha-polyglutamyl oxidized folate antimetabolite and a targeting moiety having specific affinity for an epitope of an antigen on the cancer surface. In some embodiments, the liposome comprises a targeting moiety that specifically binds to a cell surface antigen selected from: 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, CD20, CD22, CD26, CD27L, CD30, CD33, CD34, CD37, CD38, CD40, CD40L, 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 administered liposomes include a targeting moiety that specifically binds to cell surface antigens determined to originate from or be expressed on a tumor of a specific target, such as a neoantigen. In some embodiments, the administered liposome composition includes pegylated liposomes (e.g., TPLp-αPANTIFOL). In some embodiments, the administered liposome composition includes non-pegylated liposomes. In some embodiments, the liposomes of the administered liposome composition include αPANTIFOL containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomes of the administered liposome composition include an alpha-tetraglutamine oxidase antagonist. In some embodiments, the liposomes of the administered liposome composition include an alpha-pentaglutamine oxidase antagonist. In other embodiments, the liposomes of the administered liposome composition contain an alpha-hexaglutamine oxidase antagonist. In some embodiments, the liposomes contain a polyglutamine oxidase antagonist as described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, αPANTIFOL 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 one of items

[13] to

[72] 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 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.

[0045] In further embodiments, the present 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) an alpha-polyglutamate oxidase antagonist (αPANTIFOL) 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-αPANTIFOL). In some embodiments, the administered liposome composition comprises non-pegylated liposomes. In some embodiments, the liposomes of the administered liposome composition contain αPANTIFOL containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomes of the administered liposome composition contain an alpha-tetraglutamine oxidase antagonist. In some embodiments, the liposomes of the administered liposome composition contain an alpha-pentaglutamine oxidase antagonist. In other embodiments, the liposomes of the administered liposome composition contain an alpha-hexaglutamine oxidase antagonist. In some embodiments, the liposomes contain a polyglutamine oxidase antagonist as described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, αPANTIFOL 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 one of items

[13] to

[72] 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.

[0046] 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 comprising liposomes containing an alpha-polyglutamic acid oxidase antagonist (Lp-αPANTIFOL) to a subject receiving or formerly receiving cancer therapy. In some embodiments, the administered liposomal composition is PLp-αPANTIFOL, NTLp-αPANTIFOL, NTPLp-αPANTIFOL, TLp-αPANTIFOL, or TPLp-αPANTIFOL. In some embodiments, the liposomes of the administered liposomal composition comprise pegylated liposomes (e.g., PLp-αPANTIFOL, NTPLp-αPANTIFOL, or TPLp-αPANTIFOL). In some embodiments, the administered liposomal composition comprises targeted liposomes (e.g., TLp-αPANTIFOL or TPLp-αPANTIFOL). In some embodiments, the administered liposome composition comprises pegylated liposomes (e.g., TPLp-αPANTIFOL) containing a targeting moiety. In some embodiments, the liposomes of the administered liposome composition comprise an alpha-polyglutamate oxidase antagonist comprising 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomes of the administered liposome composition comprise an alpha-tetraglutamate oxidase antagonist. In some embodiments, the liposomes of the administered liposome composition comprise an alpha-pentaglutamate oxidase antagonist. In other embodiments, the liposomes of the administered liposome composition comprise an alpha-hexaglutamate oxidase antagonist. In some embodiments, the liposome composition comprises a polyglutamate oxidase antagonist as described in any one of items [1] to

[12] of the Summary of the Invention section. In some embodiments, αPANTIFOL 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 one of items

[13] to

[72] of the Summary of the Invention section.

[0047] 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 an alpha-polyglutamine oxidase antagonist (e.g., Lp-αPANTIFOL, PLp-αPANTIFOL, NTLp-αPANTIFOL, NTPLp-αPANTIFOL, TLp-αPANTIFOL, or TPLp-αPANTIFOL) 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-αPANTIFOL, NTPLp-αPANTIFOL, or TPLp-αPANTIFOL). In some embodiments, the administered liposome composition comprises targeted liposomes (e.g., TLp-αPANTIFOL or TPLp-αPANTIFOL) 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-αPANTIFOL). In some embodiments, the liposomes of the administered liposome composition comprise an alpha-polyglutamic acid folate antagonist containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomes of the administered liposome composition comprise an alpha-tetraglutamic acid folate antagonist. In some embodiments, the liposomes of the administered liposome composition contain an alpha-pentaglutamine oxidase antagonist. In other embodiments, the liposomes of the administered liposome composition contain an alpha-hexaglutamine oxidase antagonist.In some embodiments, the administered liposome composition comprises an alpha-polyglutamine oxidase antagonist described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, α-PANTIFOL is a polyglutamine oxidase antagonist described in the Summary of the Invention section. In some embodiments, the liposome composition comprises a liposome described in any one of items

[13] to

[72] in the Summary of the Invention section.

[0048] In further embodiments, the present 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 an alpha-polyglutamic acid antimetabolitic agent (e.g., Lp-αPANTIFOL, PLp-αPANTIFOL, NTLp-αPANTIFOL, NTPLp-αPANTIFOL, TLp-αPANTIFOL, or TPLp-αPANTIFOL) 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 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, and ulcerative colitis. In some embodiments, the liposome composition administered comprises pegylated liposomes (e.g., PLp-αPANTIFOL, NTPLp-αPANTIFOL, or TPLp-αPANTIFOL). In some embodiments, the administered liposome composition comprises targeted liposomes (e.g., TLp-αPANTIFOL or TPLp-αPANTIFOL) 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-αPANTIFOL) containing a targeting moiety. In some embodiments, the liposomes of the administered liposome composition comprise an alpha-polyglutamic acid folate antagonist containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomes of the administered liposome composition comprise an alpha-tetraglutamic acid folate antagonist.In some embodiments, the liposomes of the administered liposome composition contain an alpha-pentaglutamine oxidase antagonist. In other embodiments, the liposomes of the administered liposome composition contain an alpha-hexaglutamine oxidase antagonist. In some embodiments, the liposomes contain a polyglutamine oxidase antagonist as described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, αPANTIFOL 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 one of items

[13] to

[72] in the Summary of the Invention section.

[0049] 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 an alpha-polyglutamic acid oxidase antagonist (e.g., Lp-αPANTIFOL, PLp-αPANTIFOL, NTLp-αPANTIFOL, NTPLp-αPANTIFOL, TLp-αPANTIFOL, or TPLp-αPANTIFOL) 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-αPANTIFOL, NTPLp-αPANTIFOL, or TPLp-αPANTIFOL). In some embodiments, the administered liposome composition comprises targeted liposomes (e.g., TLp-αPANTIFOL or TPLp-αPANTIFOL) 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 (e.g., TPLp-αPANTIFOL) that include a targeting moiety.In some embodiments, the liposomes of the administered liposome composition contain an alpha-pentaglutamine oxidase antagonist containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the liposomes of the administered liposome composition contain an alpha-tetraglutamine oxidase antagonist. In some embodiments, the liposomes of the administered liposome composition contain an alpha-pentaglutamine oxidase antagonist. In other embodiments, the liposomes of the administered liposome composition contain an alpha-hexaglutamine oxidase antagonist. In some embodiments, the liposomes contain a polyglutamine oxidase antagonist as described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, αPANTIFOL 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 one of items

[13] to

[72] in the Summary of the Invention section.

[0050] This disclosure also provides a method for delivering an alpha-polyglutamate oxidase antagonist to a target inflammatory site, the method comprising administering to an inflammatory subject a composition comprising an alpha-polyglutamate oxidase antagonist (L-αPANTIFOL) 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-αPANTIFOL). In some embodiments, the composition to be administered comprises an alpha-polyglutamate oxidase antagonist comprising 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the administered composition comprises an alpha-tetraglutamine oxidase antagonist. In some embodiments, the administered composition comprises an alpha-pentaglutamine oxidase antagonist. In other embodiments, the administered composition comprises an alpha-hexaglutamine oxidase antagonist. In some embodiments, αPANTIFOL is a polyglutamine oxidase antagonist described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, αPANTIFOL 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 one of items

[13] to

[72] in the Summary of the Invention section.

[0051] This disclosure also provides a method for delivering an alpha-polyglutamate oxidase antagonist to tumor or cancer cells, the method comprising administering to a subject having a tumor a composition comprising an alpha-polyglutamate oxidase antagonist (L-αPANTIFOL) and a targeting moiety having specific binding affinity to an epitope on a surface antigen of tumor 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-αPANTIFOL). In some embodiments, the composition to be administered comprises an alpha-polyglutamate oxidase antagonist comprising 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the composition to be administered comprises an alpha-tetraglutamate oxidase antagonist. In some embodiments, the administered composition comprises an alpha-pentaglutamine oxidase antagonist. In other embodiments, the administered composition comprises an alpha-hexaglutamine oxidase antagonist. In some embodiments, αPANTIFOL is a polyglutamine oxidase antagonist described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, αPANTIFOL 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 one of items

[13] to

[72] in the Summary of the Invention section.

[0052] In further embodiments, the Disclosure provides a method for preparing a liposome composition comprising a liposome alpha-polyglutamine oxidase antagonist (αPANTIFOL) composition, the method comprising the steps of: forming a mixture comprising a liposome component and an α-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 alpha-polyglutamine oxidase antagonist comprises 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups. In some embodiments, the polyglutamine oxidase antagonist composition comprises an alpha-tetraglutamine oxidase antagonist. In some embodiments, the polyglutamine oxidase antagonist composition comprises an alpha-pentaglutamine oxidase antagonist. In other embodiments, the polyglutamine oxidase antagonist composition comprises an alpha-hexaglutamine oxidase antagonist. In some embodiments, αPANTIFOL is a polyglutamic acid folate antagonist described in any one of items [1] to

[12] in the Summary of the Invention section. In some embodiments, αPANTIFOL 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 one of items

[13] to

[72] in the Summary of the Invention section.

[0053] In one embodiment, the present disclosure provides a kit comprising a folate antimetabolite alpha-polyglutamate composition and / or an αPANTIFOL delivery carrier such as a liposome containing αPANTIFOL and an αPANTIFOL immune complex (e.g., ADC as described herein). [Brief explanation of the drawing]

[0054] [Figure 1-1]Figures 1A to 1R show the chemical formulas of the folate antagonist pemetrexed (Figure 1A), and representative alpha-pemetrexed polyglutamates: alpha-pemetrexed diglutamate (Figure 1B), alpha-pemetrexed triglutamate (Figures 1C and 1D), alpha-pemetrexed tetraglutamate (Figures 1E and 1F), alpha-pemetrexed pentaglutamate (Figures 1G and 1H), alpha-pemetrexed hexaglutamate (Figures 1I and 1J), alpha-pemetrexed heptaglutamate (Figures 1K and 1L), and alpha-pemetrexed octaglutamate (Figures 1M and 1N). Figures 1O to 1R show the structures of representative branched alpha-pemetrexed polyglutamates, including branched polyglutamates having a gamma-glutamyl skeleton and an alpha-glutamyl branch (Figure 1P) and branched polyglutamates having an alpha-glutamyl skeleton and a gamma-glutamyl branch (Figures 1Q and 1R). [Figure 1-2] Same as above. [Figure 1-3] Same as above. [Figure 1-4] Same as above. [Figure 1-5] Same as above. [Figure 2] Figure 2 shows the relative potency of liposomal pemetrexed alpha-L hexaglutamate (liposomal aG6) and its enantiomer, liposomal alpha-D hexaglutamate (liposomal aDG6), compared to pemetrexed, after 48 hours of exposure to cancer cell lines SW620 (CRC), HT-29 (colon cancer), HCC1806 (triple-negative breast cancer), OAW28 (ovarian cancer), H292 (NSCLC, adenocarcinoma subtype), and H2342 (NSCLC, adenocarcinoma subtype). [Figure 3]Figure 3 shows an example of the dose-response relationship, expressed as the percentage of surviving cells after 48 hours of treatment, for 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 NCI H2342 non-small cell lung cancer, adenocarcinoma subtype. Folate receptor alpha-targeted liposomes containing alpha-polyglutamate pemetrexed are predicted to successfully target NCI H2342 non-small cell lung cancer cells and reduce their survival rate. [Figure 4] Figure 4 shows an example of the 48-hour 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 (colon cancer) cells. Folate receptor alpha-targeted liposomes containing alpha-polyglutamic pemetrexed are also predicted to successfully target HT-29 (colon cancer) cells and reduce their survival rate. [Figure 5] Figure 5 shows the therapeutic effects of liposomal pemetrexed alpha-L hexaglutamate (Lps Hexa aG6), liposomal pemetrexed alpha-D hexaglutamate (Lps Hexa aDG6), and pemetrexed on HCC1806 triple-negative breast cancer cells after 48 hours of exposure. [Figure 6] Figure 6 shows the therapeutic effects of liposomal pemetrexed alpha-L hexaglutamate (Lps Hexa aG6), liposomal pemetrexed alpha-D hexaglutamate (Lps Hexa aDG6), and pemetrexed on OAW28 ovarian cancer cells after 48 hours of exposure. [Figure 7]Figure 7 shows the therapeutic effects of liposomal pemetrexed alpha-L hexaglutamate (Lps Hexa aG6) and liposomal pemetrexed alpha-D hexaglutamate (Lps Hexa aDG6) on H292 non-small cell lung cancer cells after 48 hours of exposure, compared to pemetrexed alone. [Figure 8] Figure 8 shows the therapeutic effects of liposomal pemetrexed alpha-L hexaglutamate (liposomal aG6), liposomal pemetrexed alpha-D hexaglutamate (liposomal aDG6), and pemetrexed on H292 non-small cell lung 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 aG6 formulation was superior to pemetrexed in suppressing H292 non-small cell lung cancer cells. [Figure 9] Figure 9 shows the therapeutic effects of liposomal pemetrexed alpha-L hexaglutamate (liposomal aG6), liposomal pemetrexed alpha-D hexaglutamate (liposomal aDG6), 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 aG6 formulation was superior to pemetrexed in suppressing HCC1806 triple-negative breast cancer cells. [Figure 10] Figure 10 shows the therapeutic effects on OAW28 ovarian cancer cells after 48 hours of exposure to a series of concentrations of liposomal pemetrexed alpha-L hexaglutamate (liposomal aG6), liposomal pemetrexed alpha-D hexaglutamate (liposomal aDG6), and pemetrexed. At a dose of 128 nM, pemetrexed appears to be more effective than the liposomal pemetrexed aG6 formulation, but at doses of 32 nM and 64 nM, the liposomal formulations have superior therapeutic effects to pemetrexed, and at 16 nM, the therapeutic effect of liposomal pemetrexed aG6 is similar to that of pemetrexed. [Figure 11]Figure 11 shows the toxicity of liposomal pemetrexed alpha-L hexaglutamate (liposomal aG6), liposomal pemetrexed alpha-D hexaglutamate (liposomal aDG6), and pemetrexed to differentiated human neutrophils at 64 nM, 128 nM, and 264 nM. The figure shows that liposomal pemetrexed aG6 is significantly less toxic to differentiated human neutrophils than pemetrexed. [Figure 12] Figure 12 shows the effects of liposomal pemetrexed alpha-L hexaglutamate (liposomal aG6), liposomal alpha-D hexaglutamate (liposomal aDG6), and the corresponding pemetrexed drugs on neutrophils (differentiated from CD34+ cells) after 48 hours of exposure at various dose levels ranging from 16 to 128 nM. [Figure 13] Figure 13 shows the effects on AML12 hepatocytes after 48-hour exposure to liposomal pemetrexed alpha-L hexaglutamate (liposomal aG6), liposomal pemetrexed alpha-D hexaglutamate (liposomal aDG6), and the corresponding 16 nM, 32 nM, 64 nM, and 128 nM doses of pemetrexed. Notably, none of the tested dose levels of liposomal agents appeared to be toxic to AML12 hepatocytes after treatment with liposomal pemetrexed aG6. In contrast, pemetrexed treatment resulted in approximately a 40% reduction in AML12 hepatocyte counts at all doses investigated. [Figure 14] Figure 14 shows the effects on CCD841 colon epithelial cells after 48-hour exposure to liposomal pemetrexed alpha-L hexaglutamate (liposomal aG6), liposomal pemetrexed alpha-D hexaglutamate (liposomal aDG6), 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 the number of CCD841 colon epithelial cells, compared to a reduction of approximately 20% or less after treatment with each of the liposomal compositions tested. [Figure 15] Figure 15 shows the structures of the polyglutamate folate antagonist, cisplatin (CDDP), and two possible aG6-cisplatin complexes. 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 aG6 and cisplatin upon encountering acidic lysosomes (pH 4-5) and in the presence of intracellular chloride ions. [Figure 16] Figure 16 shows the effects of liposomal aG6 treatment in mice with weekly administration of 40 mg / kg and 80 mg / kg for 4 weeks on hematological parameters: 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 17] Figure 17 shows the effects of liposomal aG6 treatment in mice with weekly administration of 40 mg / kg and 80 mg / kg for 4 weeks on hemoglobin and reticulocyte count index. There is a minimal decrease in mean hemoglobin concentration at higher dose levels. Simultaneously, there is a slight increase in mean reticulocyte count index. [Figure 18] Figure 18 shows the effects of liposomal aG6 treatment 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 of liver aminotransferases, and furthermore, no change in mean albumin levels was observed. [Figure 19] Figure 19 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 20]Figure 20 shows the results of a survival study using liposomal pemetrexed alpha-L hexaglutamate (liposomal aG6) in a xenograft model of NSCLC(H292). The survival curve for mice (10) administered 90 mg / kg of liposomal aG6 intravenously once a week for 4 weeks (90 mg / kg subcutaneously every week for 6 weeks) is shown as a solid circle. The survival curve for mice (10) administered pemetrexed (167 mg / kg intravenously every 3 weeks for 6 weeks) is shown as a solid triangle. Administration of 167 mg / kg of pemetrexed to mice is equivalent to administration of 500 mg / m2 in humans. The survival curve for control mice (10) is shown as a hollow diamond. [Figure 21] Figures 21A-F show the results of 48 hours of liposomal pemetrexed alpha-L triglutamin (LPG) treatment for 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 aG5), liposome pemetrexed alpha-L pentaglutamate (liposome aG7), and liposome pemetrexed alpha-L octaglutamate (aG6) and alpha-L dodecaglutamate (aG12) combinations (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 empty liposome control. [Modes for carrying out the invention]

[0055] Generally, this disclosure relates to novel alpha-polyglutamine oxidase folate antimetabolites compositions. The compositions represent an advance in the treatment of existing hyperproliferative diseases such as cancer. Methods for manufacturing, delivering, and using the alpha-polyglutamine oxidase folate antimetabolites compositions are also provided. The alpha-polyglutamine oxidase 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.

[0056] I. 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.

[0057] 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).

[0058] 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.

[0059] 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).

[0060] 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.

[0061] 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 comprising an ANTIFOL salt may further comprise any of various cations, e.g., Na+, Mg2+, K+, NH4+, and / or Ca2+. In certain embodiments, the salt is typically a pharmaceutically acceptable salt. In further specific embodiments, the folate antagonist salt comprises Na+. Folate antagonists typically comprise one L-gamma-glutamyl group and are therefore considered monoglutamine-oxidized for the purposes of this disclosure.

[0062] 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.

[0063] 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.

[0064] 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-gluta Minic 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-deazaiso Folic acid; N9-CHO-5-d(i)PteGlu, N9-formyl-5-deazaisofolic acid; 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 selected from their stereoisomers.

[0065] In some embodiments, the folate antagonist is a member selected from the following: aminopterin, methotrexate, larcitrexed (also known as TOMUDEX, ZD1694 (RTX)), previtrexed (also known as BGC9331; ZD9331), pemetrexed (also known as ALIMTA, LY231514), lometrexol (LMX) (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.

[0066] The terms "polyglutamate," "polyglutamine oxidation," or variations thereof refer to a composition comprising at least one chain of two or more bonded glutamyl groups. Polyglutamate chains can be linear or branched. Linear polyglutamate chains may, for example, contain glutamyl groups with alpha-carboxyl or gamma-carboxyl group bonds. Branched polyglutamate chains may contain one or more glutamyl groups with both alpha-carboxyl and gamma-carboxyl group bonds to other glutamyl groups, thereby providing branching points for the polyglutamate. Representative branched polyglutamates are shown in Figures 1O-1R. A polyglutamate chain comprises 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 group(s) of the polyglutamate chain bond to another glutamyl group via their amino groups, but not via their carboxylic acid groups.

[0067] The terms “polyglutamate oxidized folate antimetabolites,” “polyglutamate oxidized 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 αPANTIFOL(ANTIFOL-PG) herein take into account the glutamyl groups 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 hexaglutamate oxidized folate antimetabolites or folate antimetabolite hexaglutamate.

[0068] The terms “alpha-glutamyl group,” “alpha-glutamate,” and “alpha-bond” refer to a glutamyl group containing an alpha-carboxyl group bond when they relate to the bonding of a glutamyl group. In some embodiments, the alpha-bond is an amide bond between an alpha-carboxyl group of one glutamyl group and a second glutamyl group. The alpha-bond can 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.

[0069] The terms “gamma-glutamyl group,” “gamma-glutamate,” and “gamma bond” refer to a glutamyl group containing a gamma-carboxyl group bond when these relate to the bonding of a glutamyl group. As discussed herein, when a folate antagonist enters a cell, it is polyglutamate-oxidized by the enzyme folyl polygamma-glutamate synthase (FPGS), which sequentially adds L-glutamyl groups to the glutamyl groups in the folate antagonist. Thus, alpha-polyglutamate-oxidized folate antagonist compositions are not formed in cells during folate antagonist therapy. In some embodiments, the gamma bond is an amide bond between a gamma-carboxyl group of one glutamyl group and a second glutamyl group. A gamma bond can 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 bound to the folate antagonist. In some embodiments, a gamma bond refers to an amide bond of the glutamyl group of the folate antagonist. Unless otherwise specified or clearly indicated by the context, references to gamma bonds include gamma bonds of the glutamyl group of the folate antagonist.

[0070] Unless otherwise specified, the terms “alpha-polyglutamic acid folate antimetabolites,” “αPANTIFOL,” “alpha-ANTIFOL-PG,” and their repetitions are used herein to mean the same thing and refer to a polyglutamic acid folate antimetabolites composition comprising at least one glutamyl group containing an alpha linkage. For example, a pentaglutamic acid ANTIFOL composition in which the third glutamyl group has an alpha linkage, but each of the other glutamyl groups has a gamma linkage, is considered alpha-ANTIFOL-PG in this disclosure. In some embodiments, each glutamyl group of ANTIFOL-PG other than the glutamyl group of ANTIFOL has an alpha linkage (e.g., ANTIFOL-PGn where n=5, and each of G1, G2, G3, G4, and G5 has an alpha linkage). In some embodiments, each glutamyl group of ANTIFOL-PG other than the C-terminal glutamyl group(s) and the glutamyl group of the folate antagonist has an alpha linkage (for example, ANTIFOL-PG when n=5 and each of G1, G2, G3, and G4 has an alpha linkage). n In some embodiments, each glutamyl group of PMX-PG other than the C-terminal glutamyl group(s) has an alpha linkage (for example, ANTIFOL-PG when n=5 and each of the glutamyl groups of G1, G2, G3, and G4 has an alpha linkage, as in the case of a folate antimetabolite and G1, G2, G3, and G4). n ).

[0071] As used herein, the term “isolated” means a composition in a form not found in nature. Isolated alpha-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 alpha-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). Alpha-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, alpha-polyglutamine oxidized compositions are typically mixed with a pharmaceutically acceptable carrier or diluent. In some embodiments, the isolated alpha-polyglutamine oxidation composition (e.g., alpha-polyglutamate and delivery carriers such as liposomes containing alpha-polyglutamate) contains less than 1% or less than 0.1% of undesirable DNA or protein content. In some embodiments, the alpha-polyglutamate composition (e.g., alpha-polyglutamate and delivery carriers such as liposomes containing alpha-polyglutamate) is "isolated".

[0072] 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.

[0073] 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.

[0074] 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.

[0075] 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).

[0076] The term “delivery carrier” typically refers to any composition that assists, promotes, or facilitates the entry of alpha-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.

[0077] "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 (i.e., 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.

[0078] 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.

[0079] 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.

[0080] 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 αPANTIFOL 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, 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. Other types of cancers and tumors that can be treated with αPANTIFOL compositions are described herein or are known in the art.The term “metastasis” means the spread or dissemination of a tumor, cancer, or tumor to other sites, locations, regions, organs, or tissue systems within the subject, which are distinct from the primary tumor, cancer, or neoplasm. The terms “cancer,” “malignant,” “proliferative disorder,” “proliferative disorder,” and “tumor” are not mutually exclusive when used herein.

[0081] 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 a 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, 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 α-PANTIFOL composition alone or in combination with additional therapeutic agents.

[0082] 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.

[0083] 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. In some embodiments, the cancer is selected from breast cancer, 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 sarcoma, 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.

[0084] 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.

[0085] 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.

[0086] As used herein, the term “therapeutic agent” means a drug or its derivatives or prodrugs that interact with overgrowth 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.

[0087] 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.

[0088] As used herein, the term “antometabolite” means a therapeutic 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 alpha-polyglutamine oxidase antimetaboliticant compositions are used in combination with antimetaboliticants 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.

[0089] 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.

[0090] 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.

[0091] This disclosure generally relates to novel alpha-polyglutamine oxoxide antimetabolites (ANTIFOL) compositions, as well as methods for manufacturing and using these compositions to treat diseases including hyperproliferative disorders such as cancer, immune system disorders such as rheumatoid arthritis, and infectious diseases such as HIV, malaria, and schistosomiasis.

[0092] In some embodiments, this disclosure provides the following: [1] A composition comprising an alpha-polyglutamine oxoxide antimetabolitic agent, wherein at least one glutamyl group has an alpha-carboxyl group bond; [2] A composition of item [1] wherein the folate antagonist is selected from pyritrexime, pralatrexate, AG2034, GW1843, and LY309887, or their stereoisomers; [3] A composition of item [1] wherein the folate antagonist is selected from PMX, MTX, RTX, and LMX, or their stereoisomers; [4] A composition described in item [1], 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, folate antagonist; 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 described in any one of items [1] to [5], wherein the composition is as follows: (a) Each glutamyl group of a polyglutamic folate antagonist, other than the glutamyl group of the folate antagonist, has an alpha-carboxyl group bond; or (b) Two or more glutamyl groups of the polyglutamine oxoxide antimetabolites have gammacarboxyl group bonds; [7] A composition described in any one of items [1] to [5], wherein the composition is as follows: (a) Each glutamyl group other than the C-terminal glutamyl group and the glutamyl group of the folate antagonist has an alpha-carboxyl group bond; or (b) Each glutamyl group other than the C-terminal glutamyl group(s) has an alpha-carboxyl group bond; [8] A composition according to any one of items [1] to [7], wherein the alpha-polyglutamine oxoxide antimetabolitic agent is as follows: (a) Containing 4, 5, 6, 2-10, 4-6, or more than 5 glutamyl groups; (b) It is an alphapentaglutamine oxoxide antimetabolitic antagonist; or (c) It is an alpha-hexaglutamine oxoxide antimetabolite; [9] A composition according to any one of items [1] to [8], wherein the alpha-polyglutamine oxoxide antimetabolitic agent comprises 1 to 10 glutamyl groups having an alpha-carboxyl group bond;

[10] A composition described in any one of items [1] to [9], wherein the composition is as follows: (a) At least two glutamyl groups of the alpha-polyglutamine oxoxide antimetabolites are in the L-form; (b) Each glutamyl group in the alpha-polyglutamine oxoxide antimetabolites is in the L form; (c) At least one glutamyl group of the alpha-polyglutamine oxoxide antimetabolites is of the D type; (d) The glutamyl groups of each alpha-polyglutamic folate antagonist, other than the glutamyl group of the folate antagonist, are of type D; or (e) At least two glutamyl groups of the alpha-polyglutamine oxoxide antimetabolites are L-type and at least one glutamyl group is D-type;

[11] A composition according to any one of items [1] to

[10] , wherein the polyglutamate is linear;

[12] A composition according to any one of items [1] to

[10] , wherein the polyglutamate is a branched chain;

[13] A liposome composition containing an alpha-polyglutamine oxoxide antimetabolitic agent as described in any one of items [1] to

[12] (Lp-αPANTIFOL);

[14] Lp-αPANTIFOL composition of item

[13] , wherein the alpha-polyglutamine oxoxide antimetabolitic antagonist is selected from the following: (a) AG2034, pyritrexime, pralatrexate, GW1843, folate antagonists, and LY309887; or (b) PMX, MTX, RTX, and LMX, or their stereoisomers;

[15] Lp-αPANTIFOL composition of item

[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, 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-(phenysulfonyl)benzyl)ethyl)amino]quinazoline; or their stereoisomers;

[16] Lp-αPANTIFOL compositions as described in item

[13] , 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;

[17] A composition of Lp-αPANTIFOL according to any one of items

[13] to

[16] , wherein the liposomes contain an alpha-polyglutamine oxoxide antimetabolitic agent having 4, 5, 6, 2 to 10, 4 to 6, or more than 5 glutamyl groups;

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

[13] to

[17] , wherein the liposome contains an alpha-tetraglutamine oxoxide antimetabolitic agent;

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

[13] to

[17] , wherein the liposome comprises an alpha-pentaglutamine oxoxide antimetabolitic agent;

[20] Lp-αPANTIFOL composition according to any one of items

[13] to

[17] , wherein the liposome comprises an alpha-hexaglutamine oxoxide antimetabolitic agent;

[21] Lp-αPANTIFOL compositions according to any one of items

[13] to

[20] , wherein the polyglutamate is linear or branched;

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

[13] to

[21] , which: (a) Each glutamyl group other than the glutamyl group of the folate antagonist has an alpha-carboxyl group bond; or (b) A composition having two or more glutamyl groups bonded to a gammacarboxyl group;

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

[13] to

[21] , which: (a) Each glutamyl group other than the C-terminal glutamyl group and the glutamyl group of the folate antagonist has an alpha-carboxyl group bond; or (b) A composition in which each glutamyl group other than the C-terminal glutamyl group(s) has an alpha-carboxyl group bond;

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

[13] to

[23] , which: (a) At least two glutamyl groups of the alpha-polyglutamine oxoxide antimetabolites are in the L-form; (b) Each glutamyl group in the alpha-polyglutamine oxoxide antimetabolites is in the L form; (c) At least one glutamyl group of the alpha-polyglutamine oxoxide antimetabolites is of the D type; (d) The glutamyl groups of each alpha-polyglutamic folate antagonist, other than the glutamyl group of the folate antagonist, are of type D; or (e) At least two glutamyl groups of the alpha-polyglutamine oxoxide antimetabolites are L-type and at least one glutamyl group is D-type;

[25] A composition of Lp-αPANTIFOL as described in any one of items

[13] to

[24] , wherein the liposomes are pegylated (PαLp-αPANTIFOL);

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

[13] to

[24] , wherein the liposomes are not pegylated;

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

[13] to

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

[28] Lp-αPANTIFOL compositions according to any one of items

[13] to

[27] , wherein the polyglutamate is linear or branched;

[29] A Lp-αPANTIFOL composition according to any one of items

[13] to

[28] , wherein the liposome contains at least 1% by weight of an alpha-polyglutamine oxidase antagonist, or, during the process of preparing Lp-αPANTIFOL, at least 1% of a starting material of an alpha-polyglutamine oxidase antagonist is encapsulated in the Lp-αPANTIFOL;

[30] Lp-αPANTIFOL composition according to any one of items

[13] to

[29] , wherein the liposomes have a diameter in the range of 20 nm to 500 nm or 20 nm to 200 nm;

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

[13] to

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

[32] A composition of Lp-αPANTIFOL according to any one of items

[13] to

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

[33] A composition of Lp-αPANTIFOL as described in item

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

[34] Lp-αPANTIFOL compositions according to item

[32] or

[33] , 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;

[35] Lp-αPANTIFOL compositions according to any one of items

[32] to

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

[36] A composition of Lp-αPANTIFOL according to any one of items

[32] to

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

[37] A composition of Lp-αPANTIFOL as described in item

[36] , 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;

[38] A composition of Lp-αPANTIFOL as described in item

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

[39] A composition of Lp-αPANTIFOL according to any one of items

[13] to

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

[40] A composition of Lp-αPANTIFOL according to any one of items

[13] to

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

[41] An Lp-αPANTIFOL composition according to any one of items

[13] to

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

[42] Lp-αPANTIFOL composition according to any one of items

[13] to

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

[43] A composition of Lp-αPANTIFOL according to any one of items

[13] to

[38] , wherein the liposomes are cationic;

[44] A Lp-αPANTIFOL composition according to any one of items

[13] to

[43] , wherein the liposome has an internal space containing an alpha-polyglutamine oxidase antagonist and an aqueous pharmaceutically acceptable carrier;

[45] Lp-αPANTIFOL compositions of item

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

[46] Lp-αPANTIFOL composition of item

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

[47] Lp-αPANTIFOL composition of item

[46] , comprising 5% to 20% by weight of trehalose as a pharmaceutically acceptable carrier;

[48] ​​A composition of Lp-αPANTIFOL according to any one of items

[44] to

[47] , wherein the pharmaceutically acceptable carrier comprises 1% to 15% by weight of dextrose;

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

[44] to

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

[50] Lp-αPANTIFOL compositions according to any one of items

[44] to

[49] , 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;

[51] Lp-αPANTIFOL compositions according to any one of items

[44] to

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

[52] A Lp-αPANTIFOL composition according to any one of items

[13] to

[51] , 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;

[53] A Lp-αPANTIFOL composition according to any one of items

[13] to

[52] , wherein the liposomes contain less than 500,000 or less than 200,000 alpha-polyglutamine oxidase antagonist molecules;

[54] A Lp-αPANTIFOL composition according to any one of items

[13] to

[53] , wherein the liposomes contain 10 to 100,000 or any range in between alpha-polyglutamine oxidase antagonist molecules;

[55] A composition according to any one of items

[13] to

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

[56] Lp-αPANTIFOL compositions as described in item

[55] , 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;

[57] Lp-αPANTIFOL compositions of item

[55] or

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

[58] A composition of Lp-αPANTIFOL according to any one of items

[55] to

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

[59] Lp-αPANTIFOL composition according to any one of items

[55] to

[58] , wherein the targeting portion binds to a surface antigen with an equilibrium dissociation constant (Kd) in the range of 0.5 x 10⁻¹⁰ to 10 x 10⁻⁶ as measured by BIACORE® analysis;

[60] A Lp-αPANTIFOL composition according to any one of items

[55] to

[59] , wherein the targeting moiety specifically binds to one or more folate receptors selected from folate receptor alpha (FR-α), folate receptor beta (FR-β), and folate receptor delta (FR-δ);

[61] A Lp-αPANTIFOL composition according to any one of items

[55] to

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

[62] A composition of Lp-αPANTIFOL according to any one of items

[55] to

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

[63] A Lp-αPANTIFOL composition according to any one of items

[44] 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 conjugated to the PEG or outer surface of a liposome;

[64] Lp-αPANTIFOL compositions of item

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

[65] Lp-αPANTIFOL compositions of item

[63] or

[64] , wherein the immunostimulant is at least one selected from fluorescein, fluorescein isothiocyanate (FITC), DNP, beta-glucan, beta-1,3-glucan, beta-1,6-glucan, resolvin (e.g., resolvin D, resolvin E, or T-series resolvins such as Dn-6DPA or Dn-3DPA), oxidized low-density lipoprotein (e.g., OXPAC, PGPC), and Toll-like receptor (TLR) modulators such as erythritol lipids (e.g., E5564);

[66] A composition of Lp-αPANTIFOL described in any one of items

[63] to

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

[67] A composition comprising Lp-αPANTIFOL according to any one of items

[63] to

[66] , further comprising a hapten;

[68] Lp-αPANTIFOL composition of item

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

[69] A composition of Lp-αPANTIFOL according to any one of items

[13] to

[68] , further comprising at least one cryoprotective substance selected from mannitol, trehalose, sorbitol, and sucrose;

[70] Targeted compositions comprising any one of items [1] to

[69] ;

[71] Non-targeting compositions comprising any one of items [1] to

[54] and

[64] to

[69] ;

[72] A composition of Lp-αPANTIFOL according to any one of items

[13] to

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

[73] A pharmaceutical composition comprising a liposomal alpha-polyglutamine oxidized folate antimetabolitic agent composition described in any one of items

[13] to

[72] ;

[74] A pharmaceutical composition comprising an alpha-polyglutamine oxoxide antimetabolitic agent composition described in any one of items [1] to [8];

[75] A composition described in any one of items [1] to

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

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

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

[77] A method for treating or preventing a disease of which treatment or prevention is in need, comprising the step of administering a composition of any one of items [1] to

[75] to the subject;

[78] A method for treating or preventing a disease of which treatment or prevention is in need of treatment or prevention, comprising the step of administering a liposomal alpha-polyglutamine oxidized folate antimetabolitic composition described in any one of items

[13] to

[74] to the target;

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

[74] ;

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

[13] to

[74] ;

[81] A method of item

[79] or

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

[82] A method for treating cancer, comprising the step of administering an effective amount of any one of items [1] to

[74] to a subject having or at risk of having cancer;

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

[13] to

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

[84] Methods of item

[82] or

[83] 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 methods selected from, for example, hematological malignancies, including leukemia, lymphoma and other B-cell malignancies, myeloma and other plasma cell proliferation disorders;

[85] Methods of item

[82] or

[83] wherein the cancer is a member 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, or central nervous system (CNS) lymphoma;

[86] Methods of item

[82] or

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

[87] A method of item

[82] or

[83] wherein the cancer is a sarcoma such as osteosarcoma;

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

[55] to

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

[89] Maintenance therapy comprising the step of administering an effective amount of any one of items [1] to

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

[90] Maintenance therapy comprising the step of administering an effective amount of a liposomal alpha-polyglutamine oxidized folate antimetabolitic composition described in any one of items

[13] to

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

[91] A method for treating an immune system disorder comprising the step of administering an effective amount of any one of items [1] to

[74] 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;

[92] A method for treating an immune system disorder comprising the step of administering an effective amount of a liposomal alpha-polyglutamine oxidized folate antimetabolitic composition described in any one of items [9] to

[74] to a subject having or at risk of having an immune system disorder, optionally 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 arteriovenous disease, and psoriasis;

[93] Treatment methods below: (a) A method for treating an infectious disease, comprising the step of administering an effective amount of any one of items [1] to

[74] 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 one of items [1] to

[74] 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 one of items [1] to

[74] 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 one of items [1] to

[74] 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 one of items [1] to

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

[74] to a subject having or at risk of having a skin disease, wherein the skin disease is psoriasis;

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

[13] to

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

[95] A method for delivering an alpha-polyglutamine oxidase antagonist to a tumor expressing a folate receptor on its surface, comprising the step of administering an Lp-αPANTIFOL composition described in any one of items [1] to

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

[96] A method for preparing an alphapolyglutamine oxidase antagonist composition comprising a liposome alphapolyglutamine oxidase antagonist composition described in any one of items

[13] to

[74] , comprising the steps of: forming a mixture containing liposome components and an alphapolyglutamine oxidase antagonist in solution; homogenizing the mixture in solution to form liposomes; and processing the mixture to form liposomes containing an alphapolyglutamine oxidase antagonist;

[97] A method for preparing an alphapolyglutamine oxidase antagonist composition comprising a liposome alphapolyglutamine oxidase antagonist composition described in any one of items

[13] to

[74] , comprising the steps of: forming a mixture in solution containing a liposome component and an alphapolyglutamine oxidase antagonist; and processing the mixture to form liposomes containing the alphapolyglutamine oxidase antagonist;

[98] A method of item

[97] wherein the step of processing the mixture comprises the step of homogenizing the mixture in solution to form liposomes;

[99] A method for preparing a composition according to any one of items

[55] to

[74] , comprising the steps of: forming a mixture in solution containing a liposome component and an alpha-polyglutamate oxidase antagonist; homogenizing the mixture in solution to form liposomes; processing the mixture to form liposomes that encapsulate and / or contain the alpha-polyglutamate oxidase antagonist; and imparting a targeting moiety to 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);

[0100] A method for preparing a composition according to any one of items

[55] to

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

[0101] A method relating to item

[0100] , wherein the processing step includes homogenizing a mixture in solution to form liposomes;

[0102] A method according to any one of items

[99] to

[0101] , 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

[0103] A method according to any one of items

[99] to

[0102] , 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;

[0104] A method according to any one of items

[96] to

[0103] , wherein a starting material of at least 1% alpha-polyglutamine oxoxide antimetabolites is encapsulated or enclosed in Lp-αPANTIFOL.

[0093] II. Alpha-polyglutamine oxidase antagonist (α-PANTIFOL) Generally, this disclosure relates to alpha-polyglutamate oxidized folate antimetabolites (αPANTIFOL) compositions. αPANTIFOL compositions comprise at least one glutamyl group having an alpha linkage. These compositions are structurally different from L-gamma-polyglutamate oxidized folate antimetabolites (LαPANTIFOL), which are produced in cells by the enzyme folyl-polygamma-glutamate synthase (FPGS) during folate antimetabolite therapy.

[0094] In some embodiments, the αPANTIFOL 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 the folate antagonist). In some embodiments, each glutamyl group in αPANTIFOL other than the glutamyl groups of the folate antagonist has an alpha bond. In some embodiments, each glutamyl group in αPANTIFOL other than the C-terminal glutamyl group (single or multiple) and the glutamyl groups of the folate antagonist has an alpha bond. In some embodiments, each glutamyl group in αPANTIFOL other than the C-terminal glutamyl group (single or multiple) has an alpha bond. In some embodiments, two or more glutamyl groups in αPANTIFOL have gamma bonds. In some embodiments, at least one glutamyl group of the alpha-polyglutamic folate antagonist has an alpha-carboxyl group bond and a gamma-carboxyl group bond. In some embodiments, each glutamyl group in αPANTIFOL is L-type. In some embodiments, each glutamyl group in αPANTIFOL, other than the glutamyl group of the folate antagonist, is D-type. In some embodiments, αPANTIFOL contains two or more L-type glutamyl groups and one or more D-type glutamyl groups. In some embodiments, the polyglutamate chain of αPANTIFOL is linear (not branched). In some embodiments, the polyglutamate chain of αPANTIFOL is branched.

[0095] In some embodiments, the folate antagonist is selected from PMX, MTX, RTX, and LMX, or their stereoisomers.

[0096] 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. ;

[0097] In some embodiments, the folate antagonist is 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.

[0098] 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 formula (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, 6, 2 to 10, 4 to 6, or greater than 5. [ka]

[0099] 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.

[0100] 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.

[0101] 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.

[0102] 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.

[0103] 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.

[0104] 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.

[0105] 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.

[0106] 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.

[0107] In some embodiments, the folate antagonist has formula (VII), where (a) X=N; Y=NH2; and R=H; (b) X=N; Y=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.

[0108] In some embodiments, the folate antagonist is given by formula (VIII): [ka] It has the following properties: A=NH, NCH3, or CH2.

[0109] In some embodiments, the folate antagonist is given by formula (IX): [ka] The equation is such that (a) X=OH; R=H; and Y=GIu, (b) X=OCH3; R=H; and Y=GIu, (c) X=OH; R=H; and Y=valine, (d) X=OH; R=H; and Y=sverate, or (e) X=OH; R=CH3; and Y=GIu.

[0110] 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.

[0111] 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.

[0112] 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.

[0113] 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.

[0114] In other embodiments, the alpha-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.

[0115] In some embodiments, the alpha-polyglutamate folate antagonist is diglutamine-oxidized. That is, the alpha-polyglutamate folate antagonist contains one additional glutamyl group in addition to the glutamyl group in the folate antagonist (αANTIFOL-PG1), and the additional glutamyl group is bonded to the glutamyl group in the folate antagonist via an alpha bond. In some embodiments, each glutamyl group in the alpha-diglutamate folate antagonist is L-type. In other embodiments, the alpha-diglutamate folate antagonist contains a D-type glutamyl group.

[0116] In some embodiments, the alpha-polyglutamate-oxidized folate antagonist is triglutamate-oxidized. That is, the alpha-polyglutamate-oxidized folate antagonist contains two additional glutamyl groups in addition to the glutamyl group in the folate antagonist (αANTIFOL-PG2). In some embodiments, each of the two glutamyl groups has an alpha bond. In other embodiments, one of the two additional glutamyl groups has an alpha bond and the other glutamyl group has a gamma bond. In some embodiments, one of the two additional glutamyl groups has an alpha bond. In some embodiments, one of the two additional glutamyl groups has a gamma bond. In some embodiments, two of the three glutamyl groups have alpha bonds. In other embodiments, one of the three glutamyl groups has an alpha bond and the other glutamyl group has a gamma bond. In some embodiments, one glutamyl group has both an alpha bond and a gamma bond. In some embodiments, each glutamyl group of the alpha-triglutamine oxidase folate antagonist is L-type. In other embodiments, the alpha-triglutamine oxidase folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alpha-triglutamine oxidase folate antagonist, other than the glutamyl group of the folate antagonist, is D-type. In further embodiments, the triglutamine oxidase folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0117] In some embodiments, the alpha-polyglutamate-oxidized folate antagonist is tetraglutamate-oxidized and therefore contains three additional glutamyl groups in addition to the glutamyl group in the folate antagonist (αANTIFOL-PG3). In some embodiments, each of the three glutamyl groups has an alpha bond. In other embodiments, one or two of the three additional glutamyl groups have an alpha bond, and the remaining two or one glutamyl group each have a gamma bond. In some embodiments, two of the three additional glutamyl groups have an alpha bond. In other embodiments, one of the three glutamyl groups has an alpha bond, and another additional glutamyl group has a gamma bond. In other embodiments, one of the three additional glutamyl groups has both an alpha and a gamma bond. In other embodiments, three of the four glutamyl groups have an alpha bond. In some embodiments, at least one glutamyl group has both an alpha and a gamma bond. In some embodiments, the alpha-tetraglutamate folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group of the alpha-tetraglutamate folate antagonist is L-type. In other embodiments, the alpha-tetraglutamate folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alpha-tetraglutamate folate antagonist, other than the glutamyl group of the folate antagonist, is D-type. In further embodiments, the tetraglutamate folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0118] In some embodiments, the alpha-polyglutamate-oxidized folate antagonist comprises a chain of four additional glutamyl groups that are pentaglobin-oxidized (αANTIFOL-PG4) and bonded to the glutamyl group in the folate antagonist. In some embodiments, each of the four additional glutamyl groups in the chain has an alpha bond. In some embodiments, each of the four additional glutamyl groups in the chain, excluding the C-terminal glutamyl group(s), has an alpha bond. In other embodiments, one, two, or three of the four additional glutamyl groups have an alpha bond, and the remaining three, two, or one glutamyl group are each bonded to the molecule's glutamyl group via gamma bonds. In other embodiments, one or two of the four additional glutamyl groups have an alpha bond, and the remaining non-C-terminal glutamyl group is bonded to the molecule's glutamyl group via gamma bonds. In some embodiments, at least one additional glutamyl group has both an alpha and a gamma bond. In some embodiments, at least one of the five glutamyl groups has both an alpha and a gamma bond. In some embodiments, each of the five glutamyl groups in the chain, other than the C-terminal glutamyl group(s), has an alpha bond. In some embodiments, the alpha-pentaglutamine oxidase folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the alpha-pentaglutamine oxidase folate antagonist is L-type. In other embodiments, the alpha-pentaglutamine oxidase folate antagonist contains D-type glutamyl groups. In further embodiments, each glutamyl group in the alpha-pentaglutamine oxidase folate antagonist, other than the glutamyl group in the folate antagonist, is D-type. In further embodiments, the pentaglobulin oxidase folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0119] In some embodiments, the alpha-polyglutamine-oxidized folate antagonist is hexaglutamine-oxidized (αANTIFOL-PG5) and comprises a chain of five additional glutamyl groups bonded to the glutamyl group in the folate antagonist. In some embodiments, each of the five additional glutamyl groups in the chain has an alpha bond. In some embodiments, each of the five additional glutamyl groups in the chain, other than the C-terminal glutamyl group(s), has an alpha bond. In some embodiments, four of the five additional glutamyl groups in the chain have alpha bonds. In other embodiments, one, two, three, or four of the five additional glutamyl groups are bonded to the molecule's glutamyl group via alpha bonds, and the remaining four, three, two, or one glutamyl group are each bonded to the molecule's glutamyl group via gamma bonds. In other embodiments, one, two, three, or four of the five additional glutamyl groups have alpha bonds, and the remaining non-C-terminal glutamyl groups are bonded to the molecule's glutamyl groups via gamma bonds. In some embodiments, at least one additional glutamyl group has both alpha and gamma bonds. In some embodiments, at least one of the six glutamyl groups has both alpha and gamma bonds. In some embodiments, each of the six glutamyl groups in the chain other than the C-terminal glutamyl group(s) has an alpha bond. In some embodiments, five of the six glutamyl groups have alpha bonds. In some embodiments, the alpha-hexaglutamine oxidase folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the alpha-hexaglutamine oxidase folate antagonist is L-type. In other embodiments, the alpha-hexaglutamine oxidase folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alpha-hexaglutamic folate antagonist, other than the glutamyl group of the folate antagonist, is D-type. In further embodiments, the hexaglutamic folate antagonist comprises a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0120] In some embodiments, the alpha-polyglutamic folate antagonist is heptaglutamated (αANTIFOL-PG6) and therefore contains a chain of six additional glutamyl groups bonded to the glutamyl group in the folate antagonist. In some embodiments, each of the six additional glutamyl groups has an alpha bond. In some embodiments, each of the six additional glutamyl groups in the chain, excluding the C-terminal glutamyl group(s), has an alpha bond. In some embodiments, five of the six additional glutamyl groups in the chain have alpha bonds. In other embodiments, one, two, three, four, or five of the six additional glutamyl groups have alpha bonds, and the remaining five, four, three, two, or one glutamyl group each have a gamma bond. In other embodiments, one, two, three, four, or five of the six additional glutamyl groups have alpha bonds, and the remaining non-C-terminal glutamyl groups are bonded to the glutamyl group of the molecule via gamma bonds. In some embodiments, at least one additional glutamyl group has both an alpha-bond and a gamma-bond. In some embodiments, at least one of the seven glutamyl groups has both an alpha-bond and a gamma-bond. In some embodiments, each of the seven glutamyl groups in the chain, other than the C-terminal glutamyl group(s), has an alpha-bond. In some embodiments, six of the seven glutamyl groups have an alpha-bond. In some embodiments, the alpha-heptaglutamine oxidase antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the alpha-heptaglutamine oxidase antagonist is L-type. In other embodiments, the alpha-heptaglutamine oxidase antagonist contains D-type glutamyl groups. In further embodiments, each glutamyl group in the alpha-heptaglutamine oxidase antagonist, other than the glutamyl group of the folate antagonist, is D-type. In further embodiments, the heptaglutamine oxidase folate antagonist comprises a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0121] In some embodiments, the alpha-polyglutamate-oxidized folate antagonist is octaglutamine-oxidized (αANTIFOL-PG7) and therefore contains a chain of seven additional glutamyl groups bonded to the glutamyl group in the folate antagonist. In some embodiments, each of the seven additional glutamyl groups in the chain, other than the C-terminal glutamyl group (one or more), has an alpha bond. In some embodiments, six of the seven additional glutamyl groups in the chain have an alpha bond. In some embodiments, each of the seven additional glutamyl groups has an alpha bond. In other embodiments, one, two, three, four, five, or six of the seven additional glutamyl groups have an alpha bond, and the remaining six, five, four, three, two, or one glutamyl group each have a gamma bond. In other embodiments, one, two, three, four, five, or six of the seven additional glutamyl groups have an alpha bond, and the remaining non-C-terminal glutamyl group is bonded to the glutamyl group of the molecule via a gamma bond. In some embodiments, at least one additional glutamyl group has both an alpha-bond and a gamma-bond. In some embodiments, at least one of the eight glutamyl groups has both an alpha-bond and a gamma-bond. In some embodiments, each of the eight glutamyl groups other than the C-terminal glutamyl group(s) has an alpha-bond. In some embodiments, seven of the eight glutamyl groups have an alpha-bond. In some embodiments, the alpha-octaglutamine oxidase folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group of the alpha-octaglutamine oxidase folate antagonist is L-type. In other embodiments, the alpha-octaglutamine oxidase folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alpha-octaglutamine oxidase folate antagonist other than the glutamyl group of the folate antagonist is D-type. In further embodiments, the octaglutamic oxidase folate antimetabolite comprises a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0122] In some embodiments, the alpha-polyglutamate-oxidized folate antagonist comprises a chain of eight additional glutamyl groups bonded to the glutamyl group in the folate antagonist, after being nonaglutamate-oxidized (αANTIFOL-PG8). In some embodiments, each of the eight additional glutamyl groups in the chain, excluding the C-terminal glutamyl group(s), has an alpha bond. In some embodiments, seven of the eight additional glutamyl groups in the chain have an alpha bond. In some embodiments, each of the eight additional glutamyl groups has an alpha bond. In other embodiments, one, two, three, four, five, six, or seven of the eight additional glutamyl groups have an alpha bond, and the remaining seven, six, five, four, three, two, or one glutamyl group each have a gamma bond. In other embodiments, one, two, three, four, five, six, or seven of the eight additional glutamyl groups have alpha bonds, and the remaining non-C-terminal glutamyl groups are bonded to the molecule's glutamyl groups via gamma bonds. In some embodiments, at least one additional glutamyl group has both alpha and gamma bonds. In some embodiments, at least one of the nine glutamyl groups has both alpha and gamma bonds. In some embodiments, each of the nine glutamyl groups other than the C-terminal glutamyl group(s) has an alpha bond. In some embodiments, eight of the nine glutamyl groups have alpha bonds. In some embodiments, the alfanonaglutamine oxidase folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the alfanonaglutamine oxidase folate antagonist is L-type. In other embodiments, the alfanonaglutamine oxidase folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alfanonaglutamic folate antagonist, other than the glutamyl group of the folate antagonist, is D-type. In further embodiments, the nonaglutamic folate antagonist comprises a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0123] In some embodiments, the alpha-polyglutamate-oxidized folate antagonist is deca-glutamate-oxidized (αANTIFOL-PG9) (i.e., it contains a chain of nine additional glutamyl groups bonded to the glutamyl group in the folate antagonist). In some embodiments, each of the nine additional glutamyl groups has an alpha bond. In some embodiments, each of the nine additional glutamyl groups in the chain, excluding the C-terminal glutamyl group(s), has an alpha bond. In some embodiments, eight of the nine additional glutamyl groups in the chain have alpha bonds. In other embodiments, one, two, three, four, five, six, seven, or eight of the nine additional glutamyl groups have alpha bonds, and the remaining eight, seven, six, five, four, three, two, or one glutamyl group each have a gamma bond. In other embodiments, one, two, three, four, five, six, seven, or eight of the nine additional glutamyl groups have alpha bonds, and the remaining non-C-terminal glutamyl groups are bonded to the molecule's glutamyl groups via gamma bonds. In some embodiments, at least one additional glutamyl group has both alpha and gamma bonds. In some embodiments, at least one of the ten glutamyl groups has both alpha and gamma bonds. In some embodiments, each of the ten glutamyl groups other than the C-terminal glutamyl group(s) has an alpha bond. In some embodiments, nine of the ten glutamyl groups have alpha bonds. In some embodiments, the alfadecaglutamine oxidase folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group in the alfadecaglutamine oxidase folate antagonist is L-type. In other embodiments, the alfadecaglutamine oxidase folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alphadecaglutamate folate antagonist, other than the glutamyl group of the folate antagonist, is D-type. In further embodiments, the decaglutamate folate antagonist comprises a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0124] In some embodiments, the alpha-polyglutamic folate antimetabolites are undecaglutamine-oxidized (αANTIFOL-PG10). In some embodiments, each of the 10 additional glutamyl groups has an alpha bond. In some embodiments, each of the 10 additional glutamyl groups in the chain, excluding the C-terminal glutamyl group(s), has an alpha bond. In some embodiments, 9 of the 10 additional glutamyl groups in the chain have alpha bonds. In other embodiments, 1, 2, 3, 4, 5, 6, 7, 8, or 9 of the 10 additional glutamyl groups have alpha bonds, and the remaining 9, 8, 7, 6, 5, 4, 3, 2, or 1 glutamyl group each have a gamma bond. In other embodiments, 1, 2, 3, 4, 5, 6, 7, 8, or 9 of the 10 additional glutamyl groups have alpha bonds, and the remaining non-C-terminal glutamyl groups are bonded to the glutamyl groups of the molecule via gamma bonds. In some embodiments, at least one additional glutamyl group has both an alpha-bond and a gamma-bond. In some embodiments, at least one of the eleven glutamyl groups has both an alpha-bond and a gamma-bond. In some embodiments, each of the eleven glutamyl groups other than the C-terminal glutamyl group(s) has an alpha-bond. In some embodiments, 10 of the eleven glutamyl groups have an alpha-bond. In some embodiments, the alpha-unde-decaglutamine oxidase folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group of the alpha-unde-decaglutamine oxidase folate antagonist is L-type. In other embodiments, the alpha-unde-decaglutamine oxidase folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alpha-unde-decaglutamine oxidase folate antagonist other than the glutamyl group of the folate antagonist is D-type. In further embodiments, the undecaglutamate oxidase folate antagonist comprises a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0125] In some embodiments, the alpha-polyglutamine-oxidized folate antimetabolites are dodecaglutamine-oxidized (αANTIFOL-PG11). In some embodiments, each of the 11 additional glutamyl groups has an alpha bond. In some embodiments, each of the 11 additional glutamyl groups in the chain, excluding the C-terminal glutamyl group(s), has an alpha bond. In some embodiments, 10 of the 11 additional glutamyl groups in the chain have alpha bonds. In other embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 of the 11 additional glutamyl groups have alpha bonds, and the remaining 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 glutamyl group each have a gamma bond. In other embodiments, one, two, three, four, five, six, seven, eight, nine, or ten of the eleven additional glutamyl groups have alpha bonds, and the remaining non-C-terminal glutamyl groups are bonded to the molecule's glutamyl groups via gamma bonds. In some embodiments, at least one additional glutamyl group has both alpha and gamma bonds. In some embodiments, at least one of the twelve glutamyl groups has both alpha and gamma bonds. In some embodiments, each of the twelve glutamyl groups other than the C-terminal glutamyl group(s) has an alpha bond. In some embodiments, eleven of the twelve glutamyl groups have alpha bonds. In some embodiments, the alpha-dodeca-glutamine oxidase folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group of the alpha-dodeca-glutamine oxidase folate antagonist is L-type. In other embodiments, the alphadodecadaglutamine oxidase folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alphadodecadaglutamine oxidase folate antagonist, other than the glutamyl group of the folate antagonist, is D-type. In further embodiments, the dodecaglutamine oxidase folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0126] In some embodiments, the alpha-polyglutamine-oxidized folate antimetabolites are tridecaglutamine-oxidized (αANTIFOL-PG12). In some embodiments, each of the 12 additional glutamyl groups has an alpha bond. In some embodiments, each of the 12 additional glutamyl groups in the chain, excluding the C-terminal glutamyl group(s), has an alpha bond. In some embodiments, 11 of the 12 additional glutamyl groups in the chain have alpha bonds. In other embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 of the 12 additional glutamyl groups have alpha bonds, and the remaining 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 glutamyl group each have a gamma bond. In other embodiments, one, two, three, four, five, six, seven, eight, nine, ten, or eleven of the twelve additional glutamyl groups have alpha bonds, and the remaining non-C-terminal glutamyl groups are bonded to the molecule's glutamyl groups via gamma bonds. In some embodiments, at least one additional glutamyl group has both alpha and gamma bonds. In some embodiments, at least one of the thirteen glutamyl groups has both alpha and gamma bonds. In some embodiments, each of the thirteen glutamyl groups other than the C-terminal glutamyl group(s) has an alpha bond. In some embodiments, twelve of the thirteen glutamyl groups have alpha bonds. In some embodiments, the alpha-trideca-glutamine oxidase folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group of the alpha-trideca-glutamine oxidase folate antagonist is L-type. In other embodiments, the alpha-tridekaglutamine oxidase folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alpha-tridekaglutamine oxidase folate antagonist, other than the glutamyl group of the folate antagonist, is D-type. In further embodiments, the tridecaglutamine oxidase folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0127] In some embodiments, the alpha-polyglutamine-oxidized folate antimetabolites are tetradecaglutamine-oxidized (αANTIFOL-PG13). In some embodiments, each of the 13 additional glutamyl groups has an alpha bond. In some embodiments, each of the 13 additional glutamyl groups in the chain, excluding the C-terminal glutamyl group(s), has an alpha bond. In some embodiments, 12 of the 13 additional glutamyl groups in the chain have alpha bonds. In other embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 of the 13 additional glutamyl groups have alpha bonds, and the remaining 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 glutamyl group each have a gamma bond. In other embodiments, one, two, three, fourteen, five, six, seven, eight, nine, ten, eleven, or twelve of the thirteen additional glutamyl groups have alpha bonds, and the remaining non-C-terminal glutamyl groups are bonded to the molecule's glutamyl groups via gamma bonds. In some embodiments, at least one additional glutamyl group has both alpha and gamma bonds. In some embodiments, at least one of the fourteen glutamyl groups has both alpha and gamma bonds. In some embodiments, each of the fourteen glutamyl groups other than the C-terminal glutamyl group(s) has an alpha bond. In some embodiments, thirteen of the fourteen glutamyl groups have alpha bonds. In some embodiments, the alphatetradecaglutamine oxidase folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group of the alphatetradecaglutamine oxidase folate antagonist is L-type. In other embodiments, the alphatetradecaglutamine oxidase folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alphatetradecaglutamine oxidase folate antagonist, other than the glutamyl group of the folate antagonist, is D-type. In further embodiments, the tetradecaglutamine oxidase folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the polyglutamate chain is linear.In other embodiments, the polyglutamate chain is a branched chain.

[0128] In some embodiments, the alpha-polyglutamine-oxidized folate antimetabolites are pentadeca-glutamine-oxidized (αANTIFOL-PG14). In some embodiments, each of the 14 additional glutamyl groups has an alpha bond. In some embodiments, each of the 14 additional glutamyl groups in the chain, excluding the C-terminal glutamyl group(s), has an alpha bond. In some embodiments, 13 of the 14 additional glutamyl groups in the chain have alpha bonds. In other embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 of the 14 additional glutamyl groups have alpha bonds, and the remaining 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 glutamyl group each have a gamma bond. In other embodiments, one, two, three, fourteen, five, six, seven, eight, nine, ten, eleven, twelve, or thirteen of the fourteen additional glutamyl groups have alpha bonds, and the remaining non-C-terminal glutamyl groups are bonded to the molecule's glutamyl groups via gamma bonds. In some embodiments, at least one additional glutamyl group has both alpha and gamma bonds. In some embodiments, at least one of the fifteen glutamyl groups has both alpha and gamma bonds. In some embodiments, each of the fifteen glutamyl groups other than the C-terminal glutamyl group(s) has an alpha bond. In some embodiments, fourteen of the fifteen glutamyl groups have alpha bonds. In some embodiments, the alphapentadeca glutamate antimetabolite contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group of the alphapentadeca glutamate antimetabolite is L-type. In other embodiments, the alphapentadecaglutamine oxidase folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alphapentadecaglutamine oxidase folate antagonist, other than the glutamyl group of the folate antagonist, is D-type. In further embodiments, the pentadecaglutamine oxidase folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the polyglutamate chain is linear.In other embodiments, the polyglutamate chain is a branched chain.

[0129] In some embodiments, the alpha-polyglutamine-oxidized folate antimetabolites are hexadeca-glutamine-oxidized (αANTIFOL-PG15). In some embodiments, each of the 15 additional glutamyl groups has an alpha bond. In some embodiments, each of the 15 additional glutamyl groups in the chain, excluding the C-terminal glutamyl group(s), has an alpha bond. In some embodiments, 14 of the 15 additional glutamyl groups in the chain have alpha bonds. In other embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 of the 15 additional glutamyl groups have alpha bonds, and the remaining 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 glutamyl group each have a gamma bond. In other embodiments, one, two, three, fourteen, sixteen of the fifteen additional glutamyl groups have alpha bonds, and the remaining non-C-terminal glutamyl groups are bonded to the molecule's glutamyl groups via gamma bonds. In some embodiments, at least one additional glutamyl group has both alpha and gamma bonds. In some embodiments, at least one of the sixteen glutamyl groups has both alpha and gamma bonds. In some embodiments, each of the sixteen glutamyl groups other than the C-terminal glutamyl group(s) has an alpha bond. In some embodiments, fifteen of the sixteen glutamyl groups have alpha bonds. In some embodiments, the alpha-hexadeca-glutamine oxidase folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group of the alpha-hexadeca-glutamine oxidase folate antagonist is L-type. In other embodiments, the alpha-hexadeca-glutamine-oxidized folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alpha-hexadeca-glutamine-oxidized folate antagonist, other than the glutamyl group of 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. In some embodiments, the polyglutamate chain is linear.In other embodiments, the polyglutamate chain is a branched chain.

[0130] In other embodiments, the alpha-polyglutamine-oxidized folate antimetabolites are heptadeca-glutamine-oxidized (αANTIFOL-PG16). In some embodiments, each of the 16 additional glutamyl groups has an alpha bond. In some embodiments, each of the 16 additional glutamyl groups in the chain, excluding the C-terminal glutamyl group(s), has an alpha bond. In some embodiments, 15 of the 16 additional glutamyl groups in the chain have alpha bonds. In other embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 of the 16 additional glutamyl groups have alpha bonds, and the remaining 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 glutamyl group each have a gamma bond. In other embodiments, one, two, three, four, five, sixteen, seventeen, eight, nine, ten, eleven, twelve, thirteen, fourteen, or fifteen of the sixteen additional glutamyl groups have alpha bonds, and the remaining non-C-terminal glutamyl groups are bonded to the molecule's glutamyl groups via gamma bonds. In some embodiments, at least one additional glutamyl group has both alpha and gamma bonds. In some embodiments, at least one of the seventeen glutamyl groups has both alpha and gamma bonds. In some embodiments, each of the seventeen glutamyl groups other than the C-terminal glutamyl group(s) has an alpha bond. In some embodiments, sixteen of the seventeen glutamyl groups have alpha bonds. In some embodiments, the alphaheptadeca glutamate antimetabolite contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group of the alphaheptadeca glutamate antimetabolite is L-type. In other embodiments, the alphaheptadeca glutamate oxidase folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alphaheptadeca glutamate oxidase folate antagonist, other than the glutamyl group of the folate antagonist, is D-type. In further embodiments, the heptadeca glutamate oxidase folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups.In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0131] In some embodiments, the alpha-polyglutamine-oxidized folate antimetabolites are octadeca-glutamine-oxidized (αANTIFOL-PG17). In some embodiments, each of the 17 additional glutamyl groups has an alpha bond. In some embodiments, each of the 17 additional glutamyl groups in the chain, excluding the C-terminal glutamyl group(s), has an alpha bond. In some embodiments, 16 of the 17 additional glutamyl groups in the chain have alpha bonds. In other embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 of the 17 additional glutamyl groups have alpha bonds, and the remaining 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 glutamyl group each have a gamma bond. In other embodiments, one, two, three, four, five, sixteen, seventeen additional glutamyl groups have alpha bonds, and the remaining non-C-terminal glutamyl groups are bonded to the molecule's glutamyl groups via gamma bonds. In some embodiments, at least one additional glutamyl group has both alpha and gamma bonds. In some embodiments, at least one of the eighteen glutamyl groups has both alpha and gamma bonds. In some embodiments, each of the eighteen glutamyl groups other than the C-terminal glutamyl group(s) has an alpha bond. In some embodiments, seventeen of the eighteen glutamyl groups have alpha bonds. In some embodiments, the alpha-octadeca glutamate oxidase antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group of the alpha-octadeca glutamate oxidase antagonist is L-type. In other embodiments, the alpha-octadeca glutamate oxidase folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alpha-octadeca glutamate oxidase folate antagonist, other than the glutamyl group of the folate antagonist, is D-type. In further embodiments, the octadeca glutamate oxidase folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups.In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0132] In some embodiments, the alpha-polyglutamine-oxidized folate antimetabolites are nonadeca-glutamine-oxidized (αANTIFOL-PG18). In some embodiments, each of the 18 additional glutamyl groups has an alpha bond. In some embodiments, each of the 18 additional glutamyl groups in the chain, excluding the C-terminal glutamyl group(s), has an alpha bond. In some embodiments, 17 of the 18 additional glutamyl groups in the chain have alpha bonds. In other embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 of the 18 additional glutamyl groups have alpha bonds, and the remaining 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 glutamyl group each have a gamma bond. In other embodiments, one, two, three, four, five, six, seventeen, nine, ten, eleven, twelve, thirteen, fourteen additional glutamyl groups have alpha bonds, and the remaining non-C-terminal glutamyl groups are bonded to the molecule's glutamyl groups via gamma bonds. In some embodiments, at least one additional glutamyl group has both alpha and gamma bonds. In some embodiments, at least one of the nineteen glutamyl groups has both alpha and gamma bonds. In some embodiments, each of the nineteen glutamyl groups other than the C-terminal glutamyl group(s) has an alpha bond. In some embodiments, eighteen of the nineteen glutamyl groups have alpha bonds. In some embodiments, the alfanonadecaglutamine oxidase folate antagonist contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group of the alfanonadecaglutamine oxidase folate antagonist is L-type. In other embodiments, the alfanonadecaglutamine oxidase folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alfanonadecaglutamine oxidase folate antagonist, other than the glutamyl group of the folate antagonist, is D-type. In further embodiments, the nonadecaglutamine oxidase folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups.In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0133] In some embodiments, the alpha-polyglutamine-oxidized folate antimetabolites are eicosa-glutamine-oxidized (αANTIFOL-PG19). In some embodiments, each of the 19 additional glutamyl groups has an alpha bond. In some embodiments, each of the 19 additional glutamyl groups in the chain, excluding the C-terminal glutamyl group(s), has an alpha bond. In some embodiments, 18 of the 19 additional glutamyl groups in the chain have alpha bonds. In other embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 of the 19 additional glutamyl groups have alpha bonds, and the remaining 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 glutamyl group each have a gamma bond. In other embodiments, one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, three, six, fifteen, six, seven, or eighteen of the nineteen additional glutamyl groups have alpha bonds, and the remaining non-C-terminal glutamyl groups are bonded to the glutamyl groups of the molecule via gamma bonds. In some embodiments, at least one additional glutamyl group has both alpha and gamma bonds. In some embodiments, at least one of the twenty glutamyl groups has both alpha and gamma bonds. In some embodiments, each of the twenty glutamyl groups other than the C-terminal glutamyl group(s) has an alpha bond. In some embodiments, nineteen of the twenty glutamyl groups have alpha bonds. In some embodiments, the alpha-faicosa glutamate antimetabolite contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group of the alpha-faicosa-glutamate-oxidase-folate antagonist is L-type. In other embodiments, the alpha-faicosa-glutamate-oxidase-folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alpha-faicosa-glutamate-oxidase-folate antagonist, other than the glutamyl group of the folate antagonist, is D-type. In further embodiments, the icosa-glutamate-oxidase-folate antagonist contains a D-type glutamyl group and two or more L-type glutamyl groups.In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0134] In some embodiments, the alpha-polyglutamic folate antimetabolites are henicosaglutamine-oxidized (αANTIFOL-PG20). In some embodiments, each of the 20 additional glutamyl groups has an alpha bond. In some embodiments, each of the 20 additional glutamyl groups in the chain, excluding the C-terminal glutamyl group(s), has an alpha bond. In some embodiments, 19 of the 20 additional glutamyl groups in the chain have alpha bonds. In other embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 of the 20 additional glutamyl groups have alpha bonds, and the remaining 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 glutamyl group each have a gamma bond. In other embodiments, one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, three, nineteen of the twenty additional glutamyl groups have alpha bonds, and the remaining non-C-terminal glutamyl groups are bonded to the molecule's glutamyl groups via gamma bonds. In some embodiments, at least one additional glutamyl group has both alpha and gamma bonds. In some embodiments, at least one of the twenty-one glutamyl groups has both alpha and gamma bonds. In some embodiments, each of the twenty-one glutamyl groups other than the C-terminal glutamyl group(s) has an alpha bond. In some embodiments, twenty of the twenty-one glutamyl groups have alpha bonds. In some embodiments, the alpha-henicosa glutamate antimetabolite contains two or more L-type glutamyl groups. In further embodiments, each glutamyl group of the alpha-helicosa-glutamine-oxidized folate antagonist is L-type. In other embodiments, the alpha-helicosa-glutamine-oxidized folate antagonist contains a D-type glutamyl group. In further embodiments, each glutamyl group of the alpha-helicosa-glutamine-oxidized folate antagonist, other than the glutamyl group of the folate antagonist, is D-type.In further embodiments, the henicosaglutamine oxidase folate antimetabolite comprises a D-type glutamyl group and two or more L-type glutamyl groups. In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0135] In some embodiments, the alpha-polyglutamic folate antagonist contains 4 to 7 glutamyl groups bound to the folate antagonist (i.e., αANTIFOL-PGn, n=4 to 7), and each of the 4 to 7 bound glutamyl groups has an alpha linkage. In some embodiments, the alpha-polyglutamic folate antagonist contains 4 to 7 glutamyl groups bound to the folate antagonist (i.e., αANTIFOL-PGn, n=4 to 7), and each of the 4 to 7 bound glutamyl groups other than the C-terminal glutamyl group(s) has an alpha linkage. 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. In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0136] In some embodiments, the alpha-polyglutamic folate antagonist (αPANTIFOL) 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 in between, including the glutamyl group of the folate antagonist. In some embodiments, each glutamyl group in αPANTIFOL other than the glutamyl group of the folate antagonist has an alpha linkage. In some embodiments, each glutamyl group in αPANTIFOL other than the C-terminal glutamyl group(s) and the glutamyl group of the folate antagonist has an alpha linkage. In some embodiments, each glutamyl group in αPANTIFOL other than the C-terminal glutamyl group(s) has an alpha linkage. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 glutamyl groups in αPANTIFOL have alpha bonds. In some embodiments, αPANTIFOL contains L-type and D-type glutamyl groups. In further embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 glutamyl groups in αPANTIFOL have alpha bonds, and 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or 0 glutamyl groups each have gamma bonds. In some embodiments, each glutamyl group in the polyglutamate structure of the polyglutamate folate antagonist is L-type. In some embodiments, each glutamyl group in αPANTIFOL other than the glutamyl group of the folate antagonist is D-type. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 glutamyl groups in αPANTIFOL are L-type. In other embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 glutamyl groups in αPANTIFOL are D-type. In some embodiments, the polyglutamate chain is linear. In other embodiments, the polyglutamate chain is branched.

[0137] In some embodiments, the alpha-polyglutamic folate antagonist (αPANTIFOL) 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 αPANTIFOL other than the glutamyl group of the folate antagonist has an alpha linkage. In some embodiments, each glutamyl group in αPANTIFOL other than the C-terminal glutamyl group(s) and the glutamyl group of the folate antagonist has an alpha linkage. In some embodiments, each glutamyl group in αPANTIFOL other than the C-terminal glutamyl group(s) has an alpha linkage. In other embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 glutamyl groups have an alpha linkage. In some embodiments, αPANTIFOL contains two or more glutamyl groups having a gamma bond. In further embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 glutamyl groups in αPANTIFOL other than the glutamyl group of the folate antagonist. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 glutamyl groups have an alpha bond. In further embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 glutamyl groups in αPANTIFOL other than the glutamyl group of the folate antagonist have alpha bonds, and 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or 0 glutamyl groups each have gamma bonds. In some embodiments, each glutamyl group in αPANTIFOL is L-type. In some embodiments, each glutamyl group in αPANTIFOL other than the glutamyl group of 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 αPANTIFOL 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 αPANTIFOL are D-type.

[0138] In some embodiments, the alpha-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. In further embodiments, the 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 additional glutamyl groups have alpha bonds. In further embodiments, the 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 glutamyl group in the alpha-polyglutamate folate antagonist has gamma bonds. In some embodiments, at least one glutamyl group has both alpha and gamma bonds. In some embodiments, the glutamyl group in the folate antagonist has an alpha bond. In some embodiments, the glutamyl group in the folate antagonist has both alpha and gamma bonds.

[0139] 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 alpha-polyglutamate folate antagonist are L-type, D-type, or L-type and D-type. In some embodiments, each glutamyl group in the alpha-polyglutamate folate antagonist is L-type. In other embodiments, each glutamyl group in the alpha-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 alpha-polyglutamate folate antagonist are L-type, and at least one glutamyl group in the alpha-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 alpha-polyglutamate folate antagonist 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 alpha-polyglutamate folate antagonist are D-type.

[0140] In further embodiments, the alphapolyglutamic acid 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 alphapolyglutamic acid folate antagonist is L-type. In other embodiments, each glutamyl group of the alphapolyglutamic acid folate antagonist, other than the glutamyl group of the folate antagonist, is D-type. In alternative embodiments, at least two glutamyl groups in the alphapolyglutamic acid folate antagonist are L-type, and at least one glutamyl group in the alphapolyglutamic acid folate antagonist is D-type.

[0141] In further embodiments, the provided composition comprises an alpha-polyglutamic acid folate antagonist comprising 1, 2, 3, 4, 5, 6, 7, 8, 9, 1-10, or 1-20 glutamyl groups having alpha bonds. In some embodiments, the alpha-polyglutamic acid 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 alpha-polyglutamic acid 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 alpha-polyglutamic folate antimetabolites contain 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.

[0142] In other embodiments, the alpha-polyglutamine oxidase antagonist comprises at least one glutamyl group having both alpha and gamma bonds. In some embodiments, the alpha-polyglutamine oxidase antagonist comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 1 to 10, or more than 10 glutamyl groups having both alpha and gamma bonds.

[0143] In some embodiments, the alpha-polyglutamic acid folate antagonist comprises at least one glutamyl group having an alpha bond and 2, 3, 4, 5, 6, 7, 8, 9, 1-10, 1-20, or more glutamyl groups having gamma bonds. For example, in some embodiments, the alpha-polyglutamic acid folate antagonist comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 1-10 L-alpha-glutamyl group bonds and further comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 1-10 L-gamma-glutamyl group bonds. In some further embodiments, the alpha-polyglutamic acid folate antagonist comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 1 to 10 L-alpha-glutamyl group bonds, and further comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 1 to 10 D-gamma-glutamyl group bonds. In additional further embodiments, the alpha-polyglutamic acid folate antagonist comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 1 to 10 D-alpha-glutamyl group bonds, and further comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 1 to 10 D-gamma-glutamyl group bonds. In additional further embodiments, the alpha-polyglutamic acid folate antagonist comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 1 to 10 D-alpha-glutamyl group bonds, and further comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 1 to 10 D-gamma-glutamyl group bonds. In other further embodiments, the alpha-polyglutamic acid folate antagonist comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 1 to 10 D-gamma-glutamyl group bonds, and further comprises 1, 2, 3, 4, 5, 6, or 1 to 10 L-gamma-glutamyl group bonds. In other embodiments, the alpha-polyglutamic acid folate antagonist comprises at least one glutamyl group having both alpha and gamma bonds. In some embodiments, the alpha-polyglutamine oxoxide antimetabolites contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 1 to 10, or more than 10 glutamyl groups having both alpha and gamma bonds.

[0144] In some embodiments, the alpha-polyglutamic acid folate antimetabolites provided herein can accept one or more additional glutamyl groups, i.e., the compositions can act as substrates for FPGAs (folyl polyglutamic acid synthetase). Reagents, assays, and reagents for measuring the ability of alpha-polyglutamic acid folate antimetabolites 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 practice.

[0145] In some embodiments, the rate of uptake by hepatocytes of the naked alpha-PANTIFOL composition disclosed herein (e.g., alpha-PANTIFOL not bound to a delivery carrier) is significantly reduced under physiological conditions compared to the uptake rate of folate antagonists. In some embodiments, the rate of hepatocyte uptake of the naked alpha-PANTIFOL composition is 30%, 20%, 15%, or less than 10% compared to the rate of folate antagonists. In further embodiments, the rate of efflux (transport) of the alpha-PANTIFOL composition disclosed herein from hepatocytes occurs at a significantly slower rate (30%, 20%, 15%, or less than 10%) compared to folate antagonists.

[0146] In some embodiments, the alpha-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 alpha-polyglutamine-oxidized folate antagonist is a hexaglutamine-oxidized folate antagonist.

[0147] In some embodiments, the alpha-polyglutamate folate antimetabolites provided herein have lower toxic side effects than folate antimetabolites. In some embodiments, the alpha-polyglutamate folate antimetabolites provided herein are less toxic to non-overgrowth cells than folate antimetabolites. In some embodiments, the alpha-polyglutamate 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 alpha-polyglutamate folate antimetabolites are hexaglutamate folate antimetabolites.

[0148] In some embodiments, the alpha-polyglutamate folate antimetabolites provided herein have lower toxic side effects than folate antimetabolites. In some embodiments, the alpha-polyglutamate 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 alpha-polyglutamate 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 an alphapolyglutamine oxidase antagonist composition at a dose of 40 mg / kg or 80 mg / kg once weekly for four weeks. In some embodiments, the alphapolyglutamine oxidase antagonist is a hexaglutamine oxidase antagonist.

[0149] In some embodiments, treatment with the alpha-polyglutamyl oxidized folic acid antagonist composition provided herein does not induce significant hematological or liver toxic side effects in an in vivo mouse model. In some embodiments, hematological side effects are evaluated by mean neutrophils, mean white blood cells, or mean platelet count. In some embodiments, liver toxic side effects are evaluated by measuring serum aspartate aminotransferase (AST), serum alanine aminotransferase (ALT), and / or serum albumin levels. In some embodiments, the alpha-polyglutamyl oxidized folic acid antagonist composition provided herein does not significantly decrease mean neutrophils, mean white blood cells, or mean platelet count. In some embodiments, the alpha-polyglutamyl oxidized folic acid antagonist composition provided herein does not significantly increase serum aspartate aminotransferase (AST) and serum alanine aminotransferase (ALT) levels. In some embodiments, the alpha-polyglutamyl oxidized folic acid antagonist composition provided herein does not significantly reduce serum albumin levels. In some embodiments, the in vivo assay comprises administering the alpha-polyglutamyl oxidized folic acid antagonist composition at 40 mg / kg or 80 mg / kg once a week for 4 weeks. In some embodiments, the alpha-polyglutamyl oxidized folic acid antagonist is a hexaglutamyl oxidized folic acid antagonist.

[0150] In some embodiments, the alpha-polyglutamyl oxidized folic acid antagonist composition does not contain a fluorine atom. In some embodiments, the alpha-polyglutamyl oxidized folic acid antagonist composition does not contain a 4-fluoroglutamyl group.

[0151] The compositions of alpha-polyglutamylated folate metabolic antagonists (αPANTIFOL) and their uses are further described in International Application Nos. PCT / US2017 / 046666 and PCT / US2017 / 046667, and U.S. Patent Application Nos. 62 / 630,820, 62 / 627,716, 62 / 627,731, 62 / 630,671, 62 / 630,825, 62 / 630,629, 62 / 630,634, 62 / 630,728, 62 / 630,637, 62 / 630,744, 62 / 583,432, 62 / 627,714, 62 / 627,741, and 62 / 627,703, respectively. The disclosure of each of these is hereby incorporated by reference in its entirety.

[0152] A. Alpha-polyglutamylated folate metabolic antagonist analogs and derivatives The disclosure also encompasses alpha-polyglutamylated folate metabolic antagonist derivatives and analogs. The compositions and methods disclosed herein are contemplated to be applicable to derivatives or analogs of any and all known polyglutamylated folate metabolic antagonists. In some embodiments, an analog corresponds to a modified form of a folate metabolic antagonist, in which case the glutamyl group of the folate metabolic antagonist does not bind to the remainder of the folate metabolic antagonist molecule via a gamma peptide bond. In some embodiments, an analog is an isomer of a folate metabolic antagonist, in which case the glutamyl group in the folate metabolic antagonist is of the D-form. In some embodiments, the polyglutamylated form of the folate metabolic antagonist, or a polyglutamylated folate metabolic antagonist analog or derivative, is not fluorinated.

[0153] 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.

[0154] In further embodiments, the alpha-polyglutamine oxoxide antimetabolitic derivative or analog has a variant polyglutamate chain. In some embodiments, the polyglutamate chain comprises one or more natural or synthetic residues other than glutamate. In some embodiments, the polyglutamate chain comprises 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.

[0155] B.αANTIFOL-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).

[0156] The folate antimetabolite polyglutamate compositions provided herein are obtained by the following synthetic method using 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 desired length can be generated and added to a precursor of the folate antimetabolite that does not have glutamyl residues. 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 folate antimetabolite precursor binds to the peptide, and the molecule is cleaved from the resin.

[0157] C. Alpha-polyglutamine oxoxide antimetabolitic complex Surprisingly, the inventors have found that polyglutamine oxidized folate antimetabolites, such as polyglutamine oxidized pemetrexed, 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 αPANTIFOL (e.g., αPANTIFOL as disclosed herein) with a therapeutic agent or a salt or acid thereof. In some embodiments, the polyglutamine oxidized folate antimetabolite is αPANTIFOL as described in Section II, or a salt or acid thereof. In some embodiments, the Disclosure provides complexes of αPANTIFOL as described in any one of the categories [1] to

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

[13] to

[72] of the embodiments for carrying out the invention.

[0158] In further embodiments, the αPANTIFOL / therapeutic complex comprises one or more αPANTIFOLs 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 αPANTIFOL / therapeutic complex comprises one or more αPANTIFOLs containing 3-10, 3-9, 3-8, or 3-7 glutamyl groups. In other embodiments, the αPANTIFOL / therapeutic complex comprises one or more αPANTIFOLs containing 4-10, 4-9, 4-8, 4-7, 4-6, or 4-5 glutamyl groups. In a particular embodiment, the complex comprises one or more αPANTIFOLs containing 3-10 glutamyl groups. In further embodiments, the αPANTIFOL / therapeutic complex comprises one or more αPANTIFOLs containing 3 to 7 glutamyl groups. In further embodiments, the αPANTIFOL / therapeutic complex comprises one or more αPANTIFOLs containing 5 glutamyl groups. In another embodiment, the αPANTIFOL / therapeutic complex comprises one or more αPANTIFOLs 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 another embodiment, the chemotherapeutic agent is a taxane-based drug. In further embodiments, the molar ratio of αPANTIFOL / therapeutic agent in the complex is in the range of 1 to 10:1. In some embodiments, the molar ratio of αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 one of the categories

[13] -

[72] of embodiments for carrying out the invention.

[0159] In alternative embodiments, the αPANTIFOL complex comprises αPANTIFOL and cyclodextrin. In some embodiments, the αPANTIFOL complex comprises αPANTIFOL as described in any one of the categories [1] to

[12] of the embodiments for carrying out the invention. In some embodiments, the αPANTIFOL complex comprises a folate antagonist as described in Section II. In some embodiments, the molar ratio of αPANTIFOL (e.g., αPANTIFOL 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / cyclodextrin complex is encapsulated in liposomes. In some embodiments, the liposomes are Lp-αPANTIFOL as described in any one of the categories

[13] -

[72] of embodiments for carrying out the invention.

[0160] In some embodiments, the Disclosure provides compositions comprising an αPANTIFOL / platinum-based chemotherapeutic agent conjugate. In some embodiments, the conjugate comprises an αPANTIFOL described in any one of the categories [1] to

[12] of the Embodiments for Carrying Out the Invention. In some embodiments, the αPANTIFOL conjugate comprises a polyglutamic acid oxidase antagonist described in Section II. In some embodiments, the platinum-based chemotherapeutic agent is selected from cisplatin, carboplatin, and oxaliplatin, or their salts or acids. In other embodiments, the αPANTIFOL / platinum-based chemotherapeutic agent conjugate comprises an analog of cisplatin, carboplatin, or oxaliplatin, or their salts or acids. In some embodiments, the molar ratio of the αPANTIFOL / 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 the αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / platinum-based agent in the conjugate 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 αPANTIFOL / platinum-based chemotherapeutic agent in the conjugate 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / platinum-based drug conjugate is encapsulated in a liposome. In some embodiments, the liposome is Lp-αPANTIFOL as described in any one of the categories

[13] to

[72] of the embodiments for carrying out the invention.

[0161] In further embodiments, the αPANTIFOL / platinum-based chemotherapeutic agent conjugate comprises an analog of cisplatin, carboplatin, oxaliplatin, or a salt or acid thereof. In some embodiments, the conjugate comprises αPANTIFOL as described in any one of the categories [1] to

[12] of the embodiments for carrying out the invention. In some embodiments, the αPANTIFOL conjugate comprises a polyglutamic acid oxidase antagonist as described in Section II. In some embodiments, the molar ratio of αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / platinum-based analog complex is encapsulated in liposomes. In some embodiments, the liposomes are Lp-αPANTIFOL as described in any one of the categories

[13] to

[72] of the embodiments for carrying out the invention.

[0162] In further embodiments, the Disclosure provides a complex comprising αPANTIFOL and cisplatin or a salt or acid thereof. In some embodiments, the complex comprises αPANTIFOL as described in any one of the categories [1] to

[12] of the embodiments for carrying out the invention. In some embodiments, the αPANTIFOL complex comprises a folate antimetabolite as described in Section II. In some embodiments, the molar ratio of αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 one of the categories

[13] -

[72] of embodiments for carrying out the invention.

[0163] In other embodiments, the Disclosure provides a complex comprising αPANTIFOL and carboplatin or a salt or acid thereof. In some embodiments, the complex comprises αPANTIFOL as described in any one of the categories [1] to

[12] of the embodiments for carrying out the invention. In some embodiments, the αPANTIFOL complex comprises a polyglutamic acid folate antimetabolite as described in Section II of this Spec. In some embodiments, the molar ratio of αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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:(21-50), or 1:>50. In some embodiments, the αPANTIFOL / 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 one of the categories

[13] -

[72] of embodiments for carrying out the invention.

[0164] In other embodiments, the Disclosure provides a complex comprising αPANTIFOL and oxaliplatin or a salt or acid thereof. In some embodiments, the complex comprises αPANTIFOL as described in any one of the categories [1] to

[12] of the embodiments for carrying out the invention. In some embodiments, the αPANTIFOL complex comprises a polyglutamic oxyl folate antimetabolite as described in Section II. In some embodiments, the molar ratio of αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 one of the categories

[13] -

[72] of the embodiments for carrying out the invention.

[0165] In further embodiments, the Disclosure provides a conjugate comprising αPANTIFOL and a platinum-based chemotherapeutic agent ("Platin") 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 αPANTIFOL / 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 αPANTIFOL / platinum (or platinum salt or acid) in the conjugate is in the range of 1 to 20:1, or any range in between. In some embodiments, the conjugate includes αPANTIFOL as described in any one of the categories [1] to

[12] of embodiments for carrying out the invention. In some embodiments, the αPANTIFOL complex comprises a polyglutamic oxidase antagonist as described in Section II. In further embodiments, the molar ratio of αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 αPANTIFOL / 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 one of the categories

[13] -

[72] of embodiments for carrying out the invention.

[0166] In some embodiments, the Disclosure provides compositions comprising an αPANTIFOL / taxane chemotherapeutic agent (taxane) conjugate. In some embodiments, the conjugate comprises an αPANTIFOL described in any one of the categories [1] to

[12] of the embodiments for carrying out the invention. In some embodiments, the αPANTIFOL conjugate comprises a polyglutamic acid oxidase antagonist described in Section II. In some embodiments, the taxane chemotherapeutic agent is sel...

Claims

[Claim 1] A liposome composition comprising liposomes encapsulating an alpha-polyglutamine-oxidized folate antimetabolites and one or more polyglutamine-oxidized or polyglutamine-oxidized folate antimetabolites, The alpha-polyglutamine oxoxide antimetabolitic agent comprises 2 to 15 glutamyl groups having an alpha-carboxyl group bond; 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; 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-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, 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-IAHQ; 5-d(i)PteGlu, 5-deazaisofolic acid; 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; are polyglutamates of folate antimetabolites selected from these; (a) At least two of the glutamyl groups of the alpha-polyglutamine oxoxide antimetabolite are L-type, (b) Whether each of the glutamyl groups in the alpha-polyglutamine oxoxide antimetabolites is L-type, (c) At least one of the glutamyl groups of the alpha-polyglutamine oxoxide antimetabolite is of type D, (d) Each of the glutamyl groups of the alpha-polyglutamic folate antagonist, other than the glutamyl group of the folate antagonist, is of type D, or (e) At least two of the glutamyl groups of the alpha-polyglutamine oxoxide antimetabolite are L-type and at least one of the glutamyl groups is D-type; The liposomes are pegylated, have a diameter of 30 nm to 175 nm, and have a zeta potential of 0 mV or less. Liposome composition.

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