Compositions containing sulfoalkyl ether cyclodextrin and folic acid derivatives and methods of using same

Abstract

Disclosed herein includes a composition containing a folic acid or folic acid derivative; and a cyclodextrin. Some embodiments relate to a cyclodextrin inclusion complex of folic acid or folic acid derivative, wherein the composition is a lyophile.

Description

LIGAN4.054WO PATENTCOMPOSITIONS CONTAINING SULFOALKYL ETHER CYCLODEXTRIN AND FOLIC ACID DERIVATIVES AND METHODS OF USING SAMEBACKGROUNDField

[0001] The present disclosure relates to a formulation containing a folic acid derivative and a cyclodextrin. In particular, the present disclosure relates to formulations comprising a folic acid derivative and a sulfoalkyl ether cyclodextrin.Description of the Related Art

[0002] Folic acid and its derivatives play an essential role in many biological processes, notable in donating methyl groups to choline phospholipids, creatine, epinephrine, and DNA. Folates enter the body from the diet and are also synthesized by intestinal bacteria with consequent adsorption from the colon. Various folic acid racemates, dextrorotary and levorotary isomers have biological activity and therapeutic use. Folinic acid is used to treat folate defici ency and to reduce the side effects of chemotheraputic agents such as methotrexate. Additionally, folinic acid has been used in combination with drugs such as 5-fluorouracil in the treatment of various cancers. Folic acid and folate, the anion thereof are forms of the water-soluble vitamin B9 and the precursors of dihydrofolic and tetrahydrofolic acids, and of the racemates, dextrorotary and levorotary isomers, and respective anions thereof. They occur naturally in food, mostly as conjugates thereof, particularly in liver, kidneys, yeast, fruit and leafy green vegetables, and can also be taken as supplements. Tetrahydrofolates are predominantly used as 5 -formyltetrahydrofolic acid (also known as folinic acid and commercially as leucovorin) and the salts, tautomers, and zwitterionic forms thereof or as 5-methyltetrahydrofolic acid and the salts thereof, for the treatment of megaloblastic folic acid anaemia, as an antidote for increasing the compatibility of folic acid antagonists for increasing the therapeutic effect of fluorinated pyrimidines and for the treatment of autoimmune diseases such as psoriasis and rheumatoid arthritis, for increasing the compatibility of certain antiparasitic formulations, for instance trimethoprim-sulfamethoxazole, and for reducing thetoxicity of dideazatetrahydrofolates in chemotherapy. 5 -methyltetrahydrofolic acid is used in particular as a drug and as a food additive, as a vitamin preparation, for the prevention of neural tube defects, for the treatment of depressive illnesses, and for influencing the homocysteine level.

[0003] Tetrahydrofolic acids and tetrahydrofolates such as folinic acid are unstable and are susceptible to oxidation and moisture. Accordingly, incorporation of a tetrahydrofolic acid into solid oral pharmaceuticals, such as oral contraceptives, represents a big challenge from a formulation point of view. Not only should the resulting solid pharmaceutical composition exhibit a satisfactory stability upon storage, but the very manufacture of the composition itself is considered problematic as exposure to oxidizing excipients, humidity and / or open air during the manufacturing process are expected to cause degradation of the tetrahydrofolic acid and should hence be avoided. Therefore, there is a need to develop a stable formulation of folic acid derivatives, particularly folinic acid, with good bioavailability.SUMMARY

[0004] In a first aspect of the present disclosure, provided herein is a pharmaceutical composition comprising:(i) a compound having the structure of Formula (II):(II)or a pharmaceutically acceptable salt thereof or ionic form thereof, wherein: each of RA, RB, and RCis independently selected from -H, -Ci-6 alkyl, -C2-ioalkenyl, -C2-ioalkynyl, -C(O)H, and -C(O)Ci-6alkyl, orRBand RCtogether with intervening atoms form a 4-8 membered ring, optionally comprising one or more double bond; and(ii) a cyclodextrin.

[0005] In one embodiment, the ionic form may be cationic form. In another embodiment, the ionic form may be a zwitterionic form

[0006] In some embodiments, the compound of Formula (II) is a compound, or pharmaceutically acceptable salt thereof or cationic form thereof, having the structure of Formula (Il-a):(Il-a).

[0007] In some embodiments, the compound of Formula (II) is compound, or pharmaceutically acceptable salt thereof or cationic form thereof is a compound, or pharmaceutically acceptable salt thereof, having the structure of Formula (Il-b):

[0008] In some embodiments, the compound of Formula (II) is compound, or pharmaceutically acceptable salt thereof or cationic form thereof is a compound, or pharmaceutically acceptable salt thereof, having the structure of Formula (II-c):

[0009] In some embodiments, RAis -H.

[0010] In some embodiments, RBis -H, -CH3, or -C(O)H. In some embodiments RCis -H, -CH3, or -C(O)H. In some embodiments, RBand RCtogether with intervening atoms form a five membered ring.

[0011] In some embodiments, the compound of Formula (II) is selected from the group consisting of:

[0012] In some embodiments, the compound of Formula (II), or pharmaceutically acceptable salt thereof or cationic form thereof, is selected from the group consisting of:CK ^OH CK ^0H

[0013] In some embodiments, the compound of Formula (II), or a pharmaceutically acceptable salt thereof or cationic form thereof has the structure:

[0014] In some embodiments, the pharmaceutically acceptable salt of the compound of Formula (II) is a sodium, potassium, calcium, or magnesium salt. In some specific embodiments, the pharmaceutically acceptable salt of the compound of Formula (II) is a calcium salt.

[0015] In some embodiments, the cyclodextrin is a sulfoalkyl ether cyclodextrin. In some such embodiments, the cyclodextrin is a compound, or a mixture thereof, of Formula (I):or a pharmaceutically acceptable salt thereof, wherein:p is 4, 5, or 6;each Ri is independently selected from -OH and -O-(C1-C8alkylene)-SO3T; andT is selected from hydrogen or a pharmaceutically acceptable cation thereof; provided that at least one R1is -O-(C1-C8alkylene)-SO3T.

[0016] In some embodiments, at least one R1is -OCH2CH2CH2CH2SO3T.

[0017] In some embodiments, the pharmaceutically acceptable cation is sodium or potassium. In other embodiments, the pharmaceutically acceptable cation is a cation of a (Ci-C6)-alkanolamine. In some such embodiments, the alkanolamine is ethanolamine, methanolamine, 2-amino-2-methyl-l -propanol, valinol, or N-methyl-(2, 3,4,5, 6-pentahydroxy-hexyl)-amine. In some embodiments, the pharmaceutically acceptable cation is a cationic form of the compound of Formula (II), In some such embodiments, the pharmaceutically acceptable cation is cation of the compound having the structure:

[0018] In some embodiments, the composition further comprises a pH adjusting agent. In some such embodiments, the pH adjusting agent is N-methyl-(2,3,4,5,6-pentahydroxy-hexyl)-amine.

[0019] In some embodiments, the composition does not contain clopidogrel.

[0020] In some embodiments, the composition is in a solid form. In some such embodiments, the composition is formulated into a pill, capsule, granules, or tablet. In other such embodiments, the composition is formulated into a reconstitutable solid. In some embodiments, the composition is a lyophile. In some embodiments, the composition is formulated into a liquid form suitable for injection.

[0021] In some embodiments, the composition may further comprise an excipient.

[0022] In some embodiments, the composition is characterized by compound of Formula (II) being in an amount of at least 96% by weight in relation to an initial content of the compound of Formula (II) after storage in a closed container for 10 days at 40 °C and 75% relative humidity.

[0023] In some embodiments, the composition may further comprise a chemotherapeutic agent. In some such embodiments, the chemotherapeutic agent is selected from the group consisting of: methotrexate, pyrimethamine, pralatrexate, 5-fluorouracil, pyrimethamine, doxorubicin, epirubicin, idarubicin, daunorubicin, and cyclophosphamide.

[0024] In a second aspect of the present disclosure, provided herein is a method for preparing the compositions described herein, said method comprising: providing a solution containing the compound of Formula (II), or a pharmaceutically acceptable salt thereof or ionic form thereof, and cyclodextrin; and freeze-drying the solution.

[0025] In a third aspect of the present disclosure, provided herein is a method of reducing an oxidative degradation of a compound of Formula (II), comprising combining an amount of the compound of Formula (II) and an amount of cyclodextrin, wherein the amount of cyclodextrin is effective in reducing the oxidative degradation of the compound of Formula (II), In some embodiments, the molar ratio of the cyclodextrin to the folic acid or folic acid derivative is in the range of about 0.1 to about 10. In some embodiments, wherein the molar ratio of the cyclodextrin to the folic acid or folic acid derivative is about 1.0 to about 1.2.

[0026] In a fourth aspect of the present disclosure, provided herein is a method of reducing a female subject’s risk of having a miscarriage or having a fetus with a birth defect,comprising administering to the female subject in need thereof an effective amount of the compositions described herein. In some embodiments,.the birth defect is a neural tube defect, a cleft lip defect, or a cleft palate defect.

[0027] In a fifth aspect of the present disclosure, provided herein is a method of treating a deficiency or disease positively affected by the administration of both folates and reduced folates selected from the group consisting of subacute encephalitis associated with dementia, vacuolar myelopathy, premature occlusive arterial disease, severe vascular disease in infancy, severe vascular disease in childhood, progressive arterial stenosis, intermittent claudication, renovascular hypertension, ischemic cerebrovascular disease, premature retinal artery occlusion, premature retinal vein occlusion, cerebral occlusive arterial disease, occlusive peripheral arterial disease, premature death due to thromboembolic disease, premature death due to ischemic heart disease, psoriasis, celiac disease, arthritic condition, inflammation conditions, megaloblastic anaemia due to folate deficiency, intestinal malabsorption, and depressive illness, said method comprising administering to a subject in need thereof an effective amount of the compositions described herein.

[0028] In a sixth aspect of the present disclosure, provided herein is a method of reducing a female's risk of having a miscarriage, reducing a female's risk of having a fetus with a neural tube defect, reducing a female's risk of having a fetus with a cleft lip defect, reducing a female's risk of having a fetus with a cleft palate defect, said method comprising administering to the female subject in need thereof an effective amount of the compositions described herein.

[0029] In a seventh aspect of the present disclosure, provided herein is a method of reducing the side effects of a chemotherapeutic agent in a subject, said method comprising administering to the subject in need thereof an effective amount the compositions described herein. In some embodiments, the side effects are selected from the group consisting of: megaloblastic anemia, nausea, vomiting, and combinations thereof. In some embodiments, the chemotherapeutic agent is selected from the group consisting of: methotrexate, pralatrexate, 5 -fluorouracil, pyrimethamine, doxorubicin, epirubicin, idarubicin, daunorubicin, and cyclophosphamide. In some specific embodiments, the chemotherapeutic agent is 5-fluorouracil or methotrexate.

[0030] In an eighth aspect of the present disclosure, provided herein is a method of treating cancer in a subject, said method comprising administering to a subject in need thereof an effective amount the compositions described herein. In some embodiments, the method may further comprise administering one or more chemotherapeutic agent to the subject. In some embodiments, the cehmotherapeutic agent is methotrexate, pralatrexate, 5-fluorouracil, pyrimethamine, doxorubicin, epirubicin, idarubicin, daunorubicin, and cyclophosphamide. In some embodiments, the subject is a mammal. In some such embodiments, the subject is a human.

[0031] In yet another aspect of the present disclosure, provided herein is a method of treating or preventing a neurodevelopmental disorder, said method comprising administering to a subject in need thereof an effective amount of a composition described herein. In some embodiments, the neurodevelopmental disorder may be fragile X syndrome, autism spectrum disorder, attention deficit hyperactivity disorder (ADHD), pervasive developmental disorder-not otherwise specified (PDD-NOS), Rett syndrome, Asperger syndrome, Angelman syndrome, generalized anxiety disorder, social anxiety disorder, bipolar disorder, epilepsy, oppositional defiant disorder, cerebral folate deficiency, FOLR1 -related cerebral folate transport deficiency, or a sleep disorder.

[0032] In still yet another aspect of the present disclosure, provided herein is a method of reducing a female's risk of having a fetus with a neurodevelopmental disorder, said method comprising administering to the female in need thereof an effective amount of a composition described herein. In some embodiments, the neurodevelopmental disorder is fragile X syndrome, autism spectrum disorder, attention deficit hyperactivity disorder (ADHD), pervasive developmental disorder-not otherwise specified (PDD-NOS), Rett syndrome, Asperger syndrome, Angelman syndrome, generalized anxiety disorder, social anxiety disorder, bipolar disorder, epilepsy, oppositional defiant disorder, cerebral folate deficiency, FOLRl-related cerebral folate transport deficiency, or a sleep disorder. In some specific embodiments, the neurodevelopmental disorder is autism spectrum disorder.

[0033] In some embodiments, wherein said method comprises administering the composition described herein at preconception, during the first trimester of pregnancy, during the second trimester of pregnancy, or during the third trimester of pregnancy.

[0034] In some embodiments, the compositions described herein may be administered orally. In other embodiments, the compositions described herein may be administered parenterally. In yet other embodiments, the compositions described herein may be administered intravenously.BRIEF DESCRIPTION OF THE DRAWINGS

[0035] FIG. 1 shows the phase solubility of the calcium salt of 5-methyl-tetrahydrofolate in the presence of CAPTISOL®.

[0036] FIG. 2 shows the dissolution curve of six tablet formulations of 5-methyl-tetrahydrofolate and CAPTISOL® using 900 ml simulated gastric fluid (SGF) as medium, at 37 °C and 50 rpm.

[0037] FIG. 3 shows the dissolution curve of six tablet formulations of 5-methyl-tetrahydrofolate and CAPTISOL® using 900 ml simulated intestinal fluid (SIF) as medium, at 37 °C and 50 rpm.DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

[0038] The term “pharmaceutically acceptable cation” refers to cations that retain the biological effectiveness and properties of a compound and, which are not biologically or otherwise undesirable for use in a pharmaceutical composition suitable for administration to a human or mammalian subject. Examples of cation include but are not limited to sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like; particularly preferred are the ammonium, potassium, sodium, calcium and magnesium cations. Other types of cations can include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, specifically such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. Many such cations are known in the art, as described in WO 87 / 05297, Johnston et al., published September 11, 1987 (incorporated by reference herein in its entirety). In some embodiments, the pharmaceutically acceptable cation may be a drug compound or active agent. In some embodiments, the pharmaceutically acceptable cation may be the cationic form of a folic acid derivative compound described herein. For example, in some embodiments, the cationic formof a folic acid derivate including but not limited to folate-PEG-amine hydrochloride having a molecular weight of 1,000 Da, 2,000 Da, 5,000 Da, 10,000 Da or more or folinate-PEG-amine hydrochloride having a molecular weight of 1,000 Da, 20,00 Da, 5,000 Da, 10,000 Da or more. In some embodiments, the molecular weight range for folate-PEG-amine hydrochloride or folinate-PEG-amine hydrochloride may be from about 1000 Da to about 10,000 Da, from about 2,000 Da to about 10,000 Da, or from about 2,000 Da to about 5,000 Da.

[0039] “The term “pharmaceutically acceptable salt” refers to salts that retain the biological effectiveness and properties of the compounds of the preferred embodiments and, which are not biologically or otherwise undesirable. In many cases, the compounds of the preferred embodiments are capable of forming acid and / or base salts by virtue of the presence of amino and / or carboxyl groups or groups similar thereto. Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, sulfonic acids (including but not limited to methanesulfonic acid, ethane sulfonic acid, and p-toluenesulfonic acid), polyanionic sulfonic acids (including but not limited to Suramin, polystyrene sulfonic acid, sulfonic acid derivatized cyclohexaamylose, sulfonic acid derivatized cycloheptaamylose, and sulfonic acid derivatized cyclooctaamylose), salicylic acid, and the like. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like; particularly preferred are the ammonium, potassium, sodium, calcium and magnesium salts. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, specifically such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. Many such salts are known in the art, as described in World Patent Publication 87 / 05297, Johnston et al., published September 11, 1987 (incorporated by reference herein). In some embodiments, the pharmaceutically acceptable salts may be derivedfrom organic bases that are drug compounds or active agents. In some embodiments, the phar ceutically acceptable salt may may be derived from a folic acid derivative compound described herein.

[0040] As used herein, “alkyl” refers to a straight or branched hydrocarbon chain that is fully saturated (i.e., contains no double or triple bonds). The alkyl group may have 1 to 20 carbon atoms (whenever it appears herein, a numerical range such as “ 1 to 20” refers to each integer in the given range; e.g., “1 to 20 carbon atoms” means that the alkyl group may consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 20 carbon atoms, although the present definition also covers the occurrence of the term “alkyl” where no numerical range is designated). The alkyl group may also be a medium size alkyl having 1 to 9 carbon atoms. The alkyl group could also be a lower alkyl having 1 to 4 carbon atoms. The alkyl group may be designated as “Ci-4 alkyl” or similar designations. By way of example only, “Ci-4 alkyl” indicates that there are one to four carbon atoms in the alkyl chain, i.e., the alkyl chain is selected from the group consisting of methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, and t-butyl. Typical alkyl groups include, but are in no way limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertiary butyl, pentyl, hexyl, and the like.

[0041] A “sulfonyl” group refers to an “-SO2R” group in which R is selected from hydrogen, Ci-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-7 carbocyclyl, Ce-io aryl, 5-10 membered heteroaryl, and 3-10 membered heterocyclyl, as defined herein.

[0042] A "zwitterion" or “zwitterionic compound” refers to a compound having both a positive and a negative charge.

[0043] “Subject” as used herein, means a human or a non-human mammal, e g., a dog, a cat, a mouse, a rat, a cow, a sheep, a pig, a goat, a non-human primate or a bird, e.g., a chicken, as well as any other vertebrate or invertebrate.

[0044] The term “mammal” is used in its usual biological sense. Thus, it specifically includes, but is not limited to, primates, including simians (chimpanzees, apes, monkeys) and humans, cattle, horses, sheep, goats, swine, rabbits, dogs, cats, rodents, rats, mice guinea pigs, or the like.

[0045] An “effective amount” or a “therapeutically effective amount” as used herein refers to an amount of a therapeutic agent that is effective to relieve, to some extent, or to reduce the likelihood of onset of, one or more of the symptoms of a disease or condition,and includes curing a disease or condition. “Curing” means that the symptoms of a disease or condition are eliminated; however, certain long-term or permanent effects may exist even after a cure is obtained (such as extensive tissue damage).

[0046] “Treat,” “treatment,” or “treating,” as used herein refers to administering a pharmaceutical composition to a subject for prophylactic and / or therapeutic purposes. The term “prophylactic treatment” refers to treating a subject who does not yet exhibit symptoms of a disease or condition, but who is susceptible to, or otherwise at risk of, a particular disease or condition, whereby the treatment reduces the likelihood that the patient will develop the disease or condition. The term “therapeutic treatment” refers to administering treatment to a subject already suffering from a disease or condition.

[0047] The term “folic acid derivative” refers to a compound having its structure derived from a folic acid (pteroyl-L-glutamate). Examples of folic acid derivative includes but are not limited to dihydrofolate, tetrahydrofolate, 5-methyltetrahydrofolate, 5,10-methylenetetrahydrofolate, 5, 10-m ethenyltetrahydrofolate, 5,10-formiminotetrahydrofolate, 5-formyltetrahydrofolate, 10-formyltetrahydrofolate, and 10-methyltetrahydrofolate. Additional examples of folic acid derivatives include folate-PEG-amine hydrochloride having a molecular weight of 1,000 Da, 2,000 Da, 5,000 Da, 10,000 Da or more; and folinate-PEG-amine hydrochloride having a molecular weight of 1,000 Da, 2,000 Da, 5,000 Da, 10,000 Da or moreln some embodiments, the molecular weight range for folate-PEG-amine hydrochloride or folinate-PEG-amine hydrochloride may be from about 1000 Da to about 10,000 Da, from about 2,000 Da to about 10,000 Da, or from about 2,000 Da to about 5,000 Da

[0048] The term “reconstituted” refers to a solid that has been combined with a solvent to form a solution or suspension. In some embodiments, the solvent is water. In some embodiments, the solid is manufactured without forming a solution during manufacturing and then subjected to solvent for the first time during reconstitution.Cyclodextrin

[0049] The term “cyclodextrin” as used herein refers to an oligosaccharide, a-cyclodextrin, P-cyclodextrin, y-cyclodextrin, or their respective derivatives, including derivatives in which one or more hydroxy on the cyclodextrin core is replaced with an alkylether, hydroxyalkylether, or sufloalkylether substituent. Examples of cyclodextrinderivative can include but are not limited to the cyclodextrins listed in Tables Al and A2 below. Preferably, the cyclodextrin is a P-cyclodextrin, hydroxypropyl -P-cy cl odextrin, or sulfoalkyl ether P-cyclodextrin.Table AlCD derivative CD derivative CD derivative Sulfoethyl ether-a-CD (SEE- Sulfoethyl ether-P-CD Sulfoethyl ether-y-CD a-CD) (SEE-P-CD) (SEE-y-CD)Sulfopropyl ether-a-CD Sulfopropyl ether-P-CD Sulfopropyl ether-y-CD (SPE-a-CD) (SPE-P-CD) (SPE-y-CD)Sulfobutyl ether-a-CD (SPE- Sulfobutyl ether-p-CD Sulfobutyl ether-y-CD a-CD) (SBE-P-CD) (SBE-y-CD) Hydroxyethyl ether-a-CD Hydroxyethyl ether-p-CD Hydroxyethyl ether-y-CD (HEE-a-CD) (HEE-P-CD) (HEE-y-CD) Hydroxypropyl ether-a-CD Hydroxypropyl ether-P- Hydroxy propyl eth er-y-CD (HPE-a-CD) CD (HPE-P-CD) (HPE-y-CD) Hydroxypropenyl ether-a- Hydroxypropenyl ether-P- Hydroxypropenyl ether-y- CD (HPNE-a-CD) CD (HPNE-p-CD) CD (HPNE-y-CD) Hydroxybutyl ether-a-CD Hydroxybutyl ether-p-CD Hydroxybutyl ether-y-CD (HBE-a-CD) (HBE-P-CD) (HBE-y-CD) Hydroxybutenyl ether-a-CD Hydroxybutenyl ether~P- Hydroxybutenyl ether-y-CD (HBNE-a-CD) CD (HBNE-P-CD) (HBNE-y-CD) Hydroxypentenyl ether-a-CD Hydroxypentenyl ether-P- Hydroxypentenyl ether-y- (HPTNE-a-CD) CD CD(HPTNE-P-CD) (HPTNE-y-CD) Carboxymethyl-a-CD (CME- Carboxymethyl-P-CD Carboxymethyl-y-CD a-C’D) (CME-P-CD) (CME-y-CD) Carboxyethyl-a-CD (CEE-a- C arb oxy ethyl ~P -CD Carboxy ethyl -y-CD (CEE- CD) (CEE-P-CD) Y-CD)Methyl ether-CD (ME-CD) Ethyl ether-CD (EE-CD) Propyl ether-CD (PE-CD)CD derivative CD derivative CD derivativeB ethyl ether-CD (BE-CD) Ethoxy carbonyl -CD (ECPropoxycarbonyl-CD (PC- CD) CD)Methoxy carbony 1 -CD Acetate-CD (AA-CD) Propionate-CD(MC-CD) (PA-CD)Butyrate-CD (BA-CD)Table A2.Mixed ether CD derivative Mixed ether CD derivative Mixed ether CD derivative Sulfobutyl-hydroxybutyl- Sulfopropyl-hydroxybutyl- Sulfoethy 1 - hy droxybuty 1 - CD (SBE-HBE-CD) CD (SPE-HBE-CD) CD (SEE-HBE-CD) Sulfobutyl-hydroxypropyl- Sulfopropyl- Sulfoethyl-hydroxypropyl- CD (SBE-HPE-CD) hydroxypropyl-CD (SPE- CD (SEE-HPE-CD)HPE-CD)Sulfobutyl -hydroxy ethyl- Sulfopropyl-hydroxyethyl- Sulfoethyl-hydroxyethyl- CD (SBE-HEE-CD) CD (SPE-HEE-CD) CD (SEE-HEE-CD) Sulfobutyl-hydroxybutenyl- Sulfopropyl- Sulfoethyl-hydroxybutenyl- CD (SBE-HBNE-CD) hydroxybutenyl-CD (SPE- CD (SEE-HBNE-CD) HBNE-CD)Sulfobutyl-ethyl Sulfopropyl-ethyl Sulfoethyl-ethyl(SBE-EE-CD) (SPE-EE-CD) (SEE-EE-CD) Sulfobutyl-methyl Sulfopropyl-methyl Sulfoethyl-methyl (SBE-ME-CD) (SPE-ME-CD) (SEE-ME-CD) Sulfobutyl -propyl Sulfopropyl-propyl Sulfoethyl-propyl (SBE-PE-CD) (SPE-PE-CD) (SEE-PE-CD) Sulfobutyl-butyl Sulfopropyl-butyl Sulfoethyl-butyl(SBE-BE-CD) (SPE-BE-CD) (SEE-BE-CD)Mixed ether CD derivative Mixed ether CD derivative Mixed ether CD derivative Sulfobutyl -carboxy m ethyl- Sulfopropyl- Sulfoethyl-carboxymethy 1 - CD (SBE-CME-CD) carboxymethyl-CD (SPE- CD (SEE-CME-CD)CME-CD)Sulfobutyl-carboxyethyl- Sulfopropyl-carboxyethyl- Sulfoethyl -carb oxy ethyl - CD (SBE-CEE-CD) CD (SPE-CEE-CD) CD (SEE-CEE-CD) Sulfobutyl-acetate-CD Sulfopropyl-acetate-CD Sulfoethyl -acetate-CD (SBE-AA-CD) (SPE-AA-CD) (SEE-AA-CD) Sulfobutyl-propionate-CD Sulfopropyl-propionate-CD Sulfoethyl-propionate-CD (SBE-PA-CD) (SPE-PA-CD) (SEE-PA-CD) Sulfobutyl-butyrate-CD Sulfopropyl-butyrate-CD Sulfoethyl-butyrate-CD (SBE-BA-CD) (SPE-BA-CD) (SEE-BA-CD) Sulfobutyl - SulfopropylSulfoethylmethoxy carbonyl-CD methoxy carbonyl-CD methoxy carbonyl-CD (SBE-MC-CD) (SPE-MC-CD) (SEE-MC-CD) Sulfobutyl-ethoxy carbonyl- Sulfopropyl- Sulfoethyl -ethoxy carbonyl - CD (SBE-EC-CD) ethoxycarbonyl-CD (SPE- CD (SEE-EC-CD)EC-CD)Sulfobutyl- Suifopropyl- Sulfoethylpropoxycarbonyl-CD (SBE- propoxycarbonyl-CD (SPE- propoxycarbonyl -CD (SEE- PC-CD) PC-CD) PC-CD) Hydroxybutyl- Hydroxypropyl- Hydroxy ethyl- hydroxybutenyl-CD (HBE- hydroxybutenyl-CD (HPE- hy droxyb uteny 1 -CD (HEE- HBNE-CD) HBNE-CD) HBNE-CD) Hydroxybutyl-ethyl Hydroxypropyl-ethyl Hydroxy ethyl-ethyl (HBE-EE-CD) (HPE-EE-CD) (HEE-EE-CD) Hydroxybutyl-methyl Hydroxypropyl-methyl Hydroxy ethyl-methyl (HBE-ME-CD) (HPE-ME-CD) (HEE-ME-CD)Hy droxybuty 1 -pr opy 1 Hydroxypropyl -propyl Hydroxy ethyl-propyl (HBE-PE-CD) (HPE-PE-CD) (HEE-PE-CD)Mixed ether CD derivative Mixed ether CD derivative Mixed ether CD derivative Hy droxyb uty 1 -b uty 1 Hydroxypropyl -butyl Hydroxy ethyl -butyl (HBE-BE-CD) (HPE-BE-CD) (HEE-BE-CD) Hydroxybutyl- Hydroxypropyl- Hydroxy ethyl- carboxymethyl-CD carboxymethyl-CD carb oxy methyl -CD (HBE-CME-CD) (HPE-CME-CD) (HEE-CME-CD) HydroxybutylHydroxypropyl- Hydroxy ethyl- carboxy ethyl-CD carboxyethyl-CD (HPE- carboxyethyl-CD (HEE- (HBE-CEE-CD) CEE-CD) CEE-CD) Hydroxybutyl-acetate-CD Hydroxypropyl -acetate-CD Hydroxy ethyl-acetate-CD (HBE-AA-CD) (HPE-AA-CD) (HEE-AA-CD)Hy droxybutyl-propi onate- Hydroxypropyl -propionate- Hydroxy ethyl -propi on ate- CD (HBE-PA-CD) CD (HPE-PA-CD) CD (HEE-PA-CD) Hydroxybutyl-butyrate-CD Hydroxypropyl-butyrate- Hydroxy ethyl-butyrate-CD (HBE-BA-CD) CD (HPE-BA-CD) (HEE-BA-CD) Hydroxybutyl- HydroxypropylHydroxy ethylmethoxycarbonyl-CD methoxy carbonyl-CD methoxy carbonyl-CD (HBE-MC-CD) (HPE-MC-CD) (HEE-MC-CD) Hydroxybutyl- HydroxypropylHydroxy ethylethoxycarbonyl-CD (HBE- ethoxy carbonyl-CD ethoxy carb ony 1 -CDEC-CD) (HPE-EC-CD) (HEE-EC-CD) Hydroxybutyl- HydroxypropylHydroxy ethyl- propoxycarbonyl-CD propoxy carb on y 1 -CD propoxycarbonyl-CD (HBE-PC-CD) (HPE-PC-CD) (HEE-PC-CD) Hydroxybutenyl-ethyl Hydroxypropenyl-ethyl Hydroxypentenyl-ethyl (HBNE-EE-CD) (HPNE-EE-CD) (HPTNE-EE-CD) Hydroxybutenyl-methyl Hydroxypropenyl-methyl Hydroxypentenyl-methyl (HBNE-ME-CD) (HPNE-ME-CD) (HPTNE-ME-CD) Hydroxybutenyl-propyl Hy droxypropeny 1 -propyl Hydroxypentenyl-propyl (HBNE-PE-CD) (HPNE-PE-CD) (HPTNE-PE-CD)Mixed ether CD derivative Mixed ether CD derivative Mixed ether CD derivative Hydroxybutenyl-butyl Hy droxypropeny 1 -butyl Hy dr oxy p en teny 1 -buty 1 (HBNE-BE-CD) (HPNE-BE-CD) (HPTNE-BE-CD) Hydroxybutenyl- Hydroxypropenyl - Hydroxypentenyl- carboxymethyl-CD carboxymethyl-CD carboxymethyl-CD (HBNE-CME-CD) (HPNE-CME-CD) (HPTNE-CME-CD)Hy droxy buteny 1 - Hydroxypropenyl- Hydroxypentenyl- carboxy ethyl-CD carboxyethyl-CD (HPNE- carboxyethyl-CD (HPTNE- (HBNE-CEE-CD)- CEE-CD) CEE-CD) Hydroxybutenyl-acetate- Hy droxypropeny 1 -acetate- Hydroxypentenyl -acetate- CD (HBNE-AA-CD) CD (HPNE-AA-CD) CD (HPTNE-AA-CD) Hydroxybutenyl- Hydroxypropenyl - Hydroxypentenyl- propionate-CD (HBNE-PA- propionate-CD propionate-CDCD) (HPNE-PA-CD) (HPTNE-PA-CD) Hydroxybuteny 1 -butyrate- Hydroxypropenyl-butyrate- Hydroxypentenyl-butyrate- CD (HBNE-BA-CD) CD (HPNE-BA-CD) CD (HPTNE-BA-CD) Hydroxybutenyl- HydroxypropenylHydroxypentenyl- m ethoxy carb ony 1 -CD methoxy carb ony 1 -CD methoxy carb ony 1 -CD (HBNE-MC-CD) (HPNE-MC-CD) (HPTNE-MC-CD) Hydroxybutenyl- Hydroxypropenyl - Hydroxypentenyl- ethoxy carbonyl-CD ethoxy carbonyl-CD ethoxy carbonyl-CD (HBNE-EC-CD) (HPNE-EC-CD) (HPTNE-EC-CD)Hy droxy buteny 1- HydroxypropenylHydroxypentenyl- propoxycarbonyl-CD propoxy carbonyl-CD propoxycarbonyl-CD (HBNE-PC-CD) (HPNE-PC-CD) (HPTNE-PC-CD)

[0050] The terms “sulfoalkyl ether cyclodextrin” and “SAE-CD” as used herein refers to a cyclodextrin derivative containing a sulfoalkyl ether substituent, such as a (C2-6 alkylene)-SC>3H or a salt thereof. The sulfoalkyl derivative of cyclodextrin can be a single derivative or a mixture of derivatives. Since the cyclodextrin derivatives contain sulfonyl groups, they can be charged species. The sulfoalkyl ether cyclodextrin can be either substituted at least at one of the primary hydroxyl groups of cyclodextrin or they are substituted at boththe primary hydroxyl groups and at the 3 -positioned hydroxyl group. Substitution at the 2- position is also possible. Examples of sulfoalkyl ether cyclodextrin include sulfobutyl ether P cyclodextrin.

[0051] In some embodiments, the sulfoalkyl ether cyclodextrin is a compound of Formula I:or a pharmaceutically acceptable salt thereof, wherein:p is 4, 5, or 6, and each Ri is -OH or -O-(Ci-Cs alkylene)-SOsT; and each T is independently hydrogen or pharmaceutically acceptable cation, provided that at least one R1is -O-(C1-C8alkylene)-SO3T.

[0052] In some embodiments, each Ri is independently–OH or -O-(Ci-Cg alkylene)-SO3T, provided that at least one Ri is OH and at least one Ri is -O-(Ci-Cs alkylene)-SO3T, wherein T is a hydrogen or pharmaceutically acceptable cation. In some embodiments, at least one Ri is independently -OH or -O-(C4 alkylene)-SO3T. In some embodiments, at least one Ri is independently a -O-(CH2)gSO3T group, wherein g is 2 to 6, or 2 to 4. In some embodiments, at least one Ri is independently -OCH2CH2CH2SO3T or OCH2CH2CH2CH2SO3T. In some embodiments, T is H. In some embodiments, T is Na⁺. In some embodiments, each T is independently selected from an alkali metal, an alkaline earth metals, ammonium ions, and amine cations such as the, and combinations thereof. In some embodiments, each T is independently selected from Li⁺, Na⁺, K⁺, Ca+2, Mg+2, ammonium, a cation of an amine, and any combination thereof. In some embodiments, each T is independently an amine cation selected from (Ci-C6)-alkylamines, piperidine, pyrazine, (Ci-C6)-alkanolamine, ethylenediamine and (C4-Cs)-cycloalkanolamine. In some embodiments, the pharmaceutically acceptable cation may be a cation or positively charged moiety (e.g., a zwitterionic form) of the compound of Formula (II).

[0053] In some embodiments, each Ri is independently -OH or -O-(Ci-C8 alkyl), provided that at least one Ri is OH and at least one Ri is -O-(Ci-Cg alkyl). In some embodiments, each Ri is independently selected from methyl ether, ethyl ether, propyl ether, butyl ether, pentyl ethyl, and hexyl ether.

[0054] In some embodiments, each Ri is independently -OH or O-(Ci-C6 alkyl)-OH, provided that at least one Ri is OH and at least one Ri is O-(Ci-Ce alkyl)-OH. In some embodiments, at least one Ri is -O-(Ci-Ce alkyl)-OH. In some embodiments, each Ri is independently selected from hydroxymethyl ether, hydroxyethyl ether, hydroxypropyl ether, hydroxybutyl ether, hydroxypentyl ether, and hydroxyhexyl ether.

[0055] In some embodiments, a cyclodextrin derivative such as a sulfoalkyl ether cyclodextrin, an alkylated cyclodextrin, or a hydroxyalkyl ether cyclodextrin can have an average degree of substitution (ADS) of 2 to 9, 4 to 8, 4 to 7.5, 4 to 7, 4 to 6.5, 4.5 to 8, 4.5 to 7.5, 4.5 to 7, 5 to 8, 5 to 7.5, 5 to 7, 5.5 to 8, 5.5 to 7.5, 5.5 to 7, 5.5 to 6.5, 6 to 8, 6 to 7.5, 6 to 7.1, 6.5 to 7.1, 6.2 to 6.9, or 6.5 per cyclodextrin, and the remaining substituents are -H.

[0056] Some embodiments provide compositions containing a single type of cyclodextrin derivative having the structure set out in Formula (I), where the composition overall contains on the average at least 1 and up to 3n + 6 alkylsulfonic acid moieties per cyclodextrin molecule. The compositions described herein also includes compositions containing cyclodextrin derivatives having a narrow or wide range for degree of substitution and high or low degree of substitution. These combinations can be optimized as needed to provide cyclodextrins having particular properties.

[0057] Exemplary SAE-CD derivatives include SBE4-P-CD, SBE7-P-CD, SBE11 - P-CD, SBE7-y-CD and SBE5-y-CD which correspond to SAE-CD derivatives of the formula I wherein p = 5, 5, 5, 6 and 6, respectively; and there are on average 4, 7, 11, 7 and 5 sulfoalkyl ether substituents present, respectively. Other exemplary' SAE-CD derivatives include those of the formula SAEx-R-CD (Formula 2), wherein SAE is sulfomethyl ether (SME), sulfoethyl ether (SEE), sulfopropyl ether (SPE), sulfobutyl ether (SBE), sulfopentyl ether (SPtE), or sulfohexyl ether (SHE); x (average or specific degree of substitution) is 1-18, 1-21, or 1-24; R(ring structure of parent cyclodextrin) is α, β or γ, respectively; and CD is cyclodextrin. The SAE functional group includes a cationic counterion as disclosed herein or generally as used in the pharmaceutical industry for the counterion of any acidic group. Since SAE-CD is a polyanionic cyclodextrin, it can be provided in different salt forms. Suitable counterions for the SAE functional group(s) include cationic organic atoms or molecules and cationic inorganic atoms or molecules. The SAE-CD can include a single type of counterion or a mixture of different counterions. The properties of the SAE-CD can be modified by changing the identity of the counterion present. For example, a first salt form of SAE-CD can have a greater electrostatic charge than a different second salt form of SAE-CD. The calcium salt form has been found to be more electronegative than the sodium salt form. Likewise, a SAE-CD having a first degree of substitution can have a greater electrostatic charge than a second SAE-CD having a different degree of substitution.

[0058] Some embodiments provide compositions of SAE-CD where the SAE-CD is a sulfobutyl ether derivative of beta cyclodextrin (SBE-β-CD) having the following structure:wherein each R is independently –H or –(CH₂)₄-SO₃Na and the average degree of substitution with –(CH₂)₄-SO₃Na groups is between 6 and 7.1.

[0059] Methods of preparing SAE-CD derivatives are varied but generally include the general steps of sulfoalkylation followed by isolation. The chemical property profile of theSAE-CD is established during the sulfoalkylation step. For example, altering reaction conditions during sulfoalkylation can vary the average degree of substitution for and the average regiochemical distribution of sulfoalkyl groups in the SAE-CD. The alkyl chain length of the sulfoalkyl functional group is determined according the sulfoalkylating agent used. And use of a particular alkalizing agent during alkylation would result in formation of a particular SAE-CD salt, unless an ion exchange step were performed subsequent to sulfoalkylation.

[0060] In general, known processes for the sulfoalkylation step include, for example: 1) exposure of underivatized parent cyclodextrin under alkaline conditions to an alkylating agent, e.g. alkyl sultone or a haloalkyl sulfonate; 2) optional addition of further alkalizing agent to the reaction milieu to consume excess alkylating agent; and 3) neutralization of the reaction medium with acidifying agent. The vast majority of literature processes conduct the sulfoalkylation step in aqueous media; however, some references disclose the use of pyridine, dioxane, or DMSO as the reaction solvent for sulfoalkylation. Literature discloses the use of an alkalizing agent in order to accelerate the sulfoalkylation reaction.

[0061] Upon completion of the sulfoalkylation step, isolation and purification of the SAE-CD is conducted.

[0062] Several different isolation processes for SAE-CD following sulfoalkylation and neutralization are described. In general, an aqueous liquid containing SAE-CD is dried to remove water to form a solid. The literature suggests various methods for removal of water from an aqueous solution containing SAE-CD. Such methods include conventional freeze-drying, spray drying, oven drying, vacuum oven drying, roto-evaporation under reduced pressure, vacuum drying or vacuum drum drying. See, for example, Ma (S. T. P. Pharma. Sciences (1999), 9(3), 261-266), CAPTISOL® (sulfobutyl ether beta-cyclodextrin sodium; Pharmaceutical Excipients 2004; Eds. R. C. Rowe, P. J. Sheskey, S. C. Owen; Pharmaceutical Press and American Pharmaceutical Association, 2004) and other references regarding the preparation of SAE-CD derivatives. In some embodiments, SAE-CD may be isolated using methods described in U. S. Patent Nos. 10,973,780 and 11,534,454, the entire disclosures of which are hereby incorporated by reference. In some embodiments, the SAE-CD may be purified based on methods described, e.g., in International Patent Publication Nos. WO 2009 / 134347, WO 2013 / 130666, WO 2014 / 066274, WO 2021 / 101842, WO 2022 / 066958, the entire disclosures of which are hereby incorporated by reference.

[0063] Suitable methods for preparing a SAE-CD- raw material for use in preparing the SAE-CD composition for use as described herein are disclosed, e.g., in U. S. Patents No.5,376,645, No. 5,874,418, No. 5,134,127, and No. 7,625,878 to Stella et al.; U. S. Patent No.3,426,011 to Parmerter et al.; Lammers et al. (Reel, Trav. CMm. Pays-Bas (1972), 91(6), 733-742); Staerke (1971), 23(5), 167-171); Qu et al. (J Inclusion Phenom. Macro. Chem., (2002), 43, 213-221); U. S. Patent No. 5,241,059 to Yoshinaga; U. S Patent No. 6,153,746 to Shah; PCT International Publication No. WO 2005 / 042584 to Stella et al; Adam et al. (J. Med. Chem. (2002), 45, 1806-1816); PCT International Publication No. WO 01 / 40316 to Zhang et al.; Tarver et al. (Bioorganic & Medicinal Chemistry (2002), 10, 1819-1827); Ma (S. T. P. Pharma. Sciences (1999), 9(3), 261-266); Jung et al. (J Chromat. 1996, 755, 81- 88); and Luna et al. (Carbohydr, Res. 1997, 299, 103-110), the entire disclosures of which are hereby incorporated by reference.

[0064] The SAE-CD raw material can be included in the liquid feed used in the fluidized bed spray drying process as described in US Patent No. 8,049,003, which is incorporated by reference for the purpose of preparing the SAE-CD composition through the fluidized bed spray drying process. Other methods for removal of water from an aqueous solution containing SAE-CD can include conventional freeze-drying, spray drying, oven drying, vacuum oven drying, roto-evaporation under reduced pressure, vacuum drying or vacuum drum drying. See, for example, Ma (S. T. P. Pharma. Sciences (1999), 9(3), 261-266), CAPTISOL®(sulfobutyl ether beta-cyclodextrin sodium; Pharmaceutical Excipients 2004; Eds. R. C. Rowe, P. J. Sheskey, S. C. Owen; Pharmaceutical Press and American Pharmaceutical Association, 2004), which is incorporated herein by reference in its entirety, and other references regarding the preparation of SAE-CD derivatives.

[0065] The SAE-CD composition described herein can also include a combination of derivatized cyclodextrin (SAE-CD) and underivatized cyclodextrin. For example, a SAE- CD composition can be made to include underivatized cyclodextrin in the amount of 0 to less than 50% by wt. of the total cyclodextrin present. Exemplary embodiments of the SAE-CD composition include those comprising 0-5% by wt., 5-50% by wt., less than 5%, less than 10%, less than 20%, less than 30%, less than 40%, or less than 50% underivatized cyclodextrin.

[0066] In some embodiments, the SAE-CD composition can also include a combination of various acid and salt forms. For example, in some embodiments, the SAE-CDcomposition may be a combination of SAE-CDs that includes both the free acid and salt forms of the SAE-CD. In some embodiments, the SAE-CD composition may be prepared by combining the SAE-CD in free acid form (i.e., wherein T is hydrogen) is combined with the compound of Formula (II) to form salts. In some embodiments, the cationic form of the compound of formula II may interaction with an anionic SAE-CD (e.g., Captisol®) to form salts.Folic Acid Derivatives

[0067] The folic acid derivative can include various compounds having structures derived from folic acid. In some embodiments, the folic acid derivative can be a tetrahydrofolic acid or pharmaceutically acceptable salt thereof. In some embodiments, the folic acid derivative can be a compound having the structure of Formula (II):(II)or a pharmaceutically acceptable salt, tautomer, or zwitterionic form thereof, wherein:each of RA, RB, and RCis independently selected from -H, -Ci-6 alkyl, -C2-ioalkenyl, -C2-ioalkynyl, -C(O)H, and -C(O)Ci-ealkyl, orRBand RCtogether with intervening atoms form a 4-8 membered ring, optionally comprising one or more double bond.

[0068] Compounds of Formula (II) include stereoisomers or racemic mixtures unless otherwise indicated.

[0069] In some embodiments, RAis H. In some embodiments, RBis H. In some embodiments, RCis H. In some embodiments, RBis methyl. In some embodiments, RBis -C(O)H. In some embodiments, RBand RCtogether with intervening atoms form a five membered ring. In some embodiments, the five membered ring is imidazolidine.

[0070] In some embodiments, the folic acid derivative of Formula (II) can be a compound, or pharmaceutically acceptable salt thereof, having the structure of Formula (Il-a):(Il-a).

[0071] In some embodiments, the folic acid derivative of Formula (II) can be a compound, or pharmaceutically acceptable salt, tautomer, or zwitterionic form thereof, having the structure of Formula (Il-b):

[0072] In some embodiments, the folic acid derivative of Formula (II) can be a compound, or pharmaceutically acceptable salt, tautomer, or zwitterionic form thereof, having the structure of Formula (II-c):(II-c).

[0073] In some embodiments, the folic acid derivative can be a compound, or pharmaceutically acceptable salt, tautomer, or zwitterionic form thereof, having the structure selected from the group consisting of:

[0074] In some embodiments, the folic acid derivative can be a compound, or pharmaceutically acceptable salt, tautomer, or zwitterionic form thereof, having the structure selected from the group consisting of:6Z~HO^oCK ^OH CK ^0Hcompound of Formula (II) can be an alkaline metal salt or an earth metal salt. In some embodiments, the pharmaceutically acceptable salt salt of the compound of Formula (II) may be a sodium, lithium, or potassium salt. In other embodiments, the pharmaceutically acceptable salt of the compound of Formula (II) may be a calcium salt or a magnesium salt.

[0076] In some embodiments, the folic acid derivative can have the structure:or a pro-drug or metabolite thereof.

[0077] In some embodiments, the folic acid derivative can have the structure:Ca2+or a stereoisomer thereof.Compositions Containing Folic Acid or Folic Acid Derivative and Sulfoalkyl Ether Cyclodextrin

[0078] Some embodiments relate to a composition including a folic acid or folic acid derivative; and a cyclodextrin.

[0079] Some embodiments relate to a composition including a cyclodextrin inclusion complex of folic acid or folic acid derivative, wherein the composition is a lyophile.

[0080] In some embodiments, the composition is characterized by the folic acid or folic acid derivative being in an amount of at least about 96% by weight in relation to the initial content of the folic acid or folic acid derivative after storage in a closed container for 10 days at 40 °C and 75% relative humidity. In some embodiments, the composition is characterized by the folic acid or folic acid derivative being in an amount of at least about 80%, 85%, 90%, 92%, 94%, 95%, 97%, 98%, or 99%, by weight in relation to the initial content of the folic acid or folic acid derivative after storage in a closed container for 10 days at 40 °C and 75% relative humidity. In some embodiments, the composition is characterized by a degradation rate of the folic acid or folic acid derivative being less than about 20%, 15%, 10%, 8%, 6%, 5%, 4%, 3%, 2%, or 1 %, by weight after storage in a closed container for 10 days at 40 °C and 75% relative humidity.

[0081] In some embodiments, the composition described herein does not contain clopidogrel.

[0082] The composition described herein can include acceptable carriers and / or auxiliary agents necessary for the administration of the composition in the desired manner. Among the carriers and / or auxiliary agents are included excipients, thickeners, diluents,solvents, dispersants or adjuvants known to the expert of the art. Thickeners include, but are not limited to, water-soluble polymers such as those selected from the group consisting of modified celluloses, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxyethylmethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose and carboxymethylcellulose, dextrans, gelatins, collagen, polyethylene glycol or polyvinyl pyrrolidone. Diluents and solvents include, but are not limited to, those selected from the group consisting of ethanol, polyethylene glycol, glycofurol, Nmethyl-2-pyrrolidone, glycerol, propanediol, polypropylene glycol, benzyl alcohol or dimethylsulfoxide. Dispersants include, but are not limited to, surfactants selected from the group consisting of monoesters of fatty acids of polyoxyethylene sorbitan (Tween®, Enialex, Nikkol®, Hodag, Dacol or Liposorb®), fatty acid monoesters of sorbitan (Span®), 15-hydroxystearate polyethylene glycol (Solutol® HS15), fatty acid esters of polyethylene glycol (Crodet, Cithrol, Kessco®, Nikkol®, Mapeg®, Myrj, Tagat®, Aldo®, Capmul®, Glycerox, Lactomul®, or Emerest®), esters of glycol polyoxyethylene (Emulphor®), poly ethoxylated castor oils (Cremophor®, Emalex, Eumulgin®, Nikkol® or Simusol®), fatty acid esters of polyglycerol (Nikkol Decaglyn, Polymuls, Caprol®), polyethylene glycol ethers (Volpo or Brij®), poloxamer (Lutrol® or Pluronic®), phenyl ethers of polyoxyethylene (Triton® or Igepal®), or mixtures thereof. In some embodiments, the cosmetic or pharmaceutical composition described herein also contains one or more acceptable excipients such as humectants, pH buffers, preservatives, bactericidal and fungicidal agents, absorption retardants, absorption accelerators, or any other excipient known to the expert of the art.

[0083] The form of the composition can vary depending on the use of the composition. The composition can be in a solid or liquid form. In some embodiments, the composition described herein is in a solid form. When a dietary supplement composition is prepared, the composition can be in the form of a pill, capsule, tablet, soft gel, liquid suspension, syrup, or powder. In some embodiments, the composition is formulated into a pill, capsule, granules, or tablet. In other embodiments, the composition is formulated into a reconstitutable solid suitable for injection. In other embodiments, the composition can be in the form of a solution or suspension for injection.

[0084] The solid pharmaceutical composition described herein can contain one or more pharmaceutically acceptable excipients. These excipients may, for example, be inertdiluents or fillers, such as sucrose, sorbitol, sugars, mannitol, microcrystalline cellulose, starches, sodium chloride, sodium phosphate, calcium carbonate, calcium phosphate, calcium sulfate, lactose, e.g. lactose monohydrate, or a combination thereof. The inert diluent or filler is typically present in an amount from 10-99% by weight of the composition. Preferably, the inert diluent or filler is present in an amount from 50-99% by weight of the composition, more preferably in an amount from 75-99% by weight of the composition, even more preferably in an amount from 80-97% by weight of the composition, most preferably in an amount from 85-97% by weight of the composition. In some embodiments, the inert fillers can be lactose, in particular lactose monohydrate, and microcrystalline cellulose. Examples of other pharmaceutically acceptable excipients which may be incorporated in the solid pharmaceutical composition described herein can also include colorants, flavoring agents, plasticizers, humectants, buffering agents, etc.

[0085] In some embodiments, the composition described herein includes lactose monohydrate, microcrystalline cellulose or a combination of lactose monohydrate and microcrystalline cellulose in the amounts indicated supra. In some embodiments, the composition described herein includes microcrystalline cellulose. The microcrystalline cellulose is typically present in an amount from 10-99% by weight of the composition, preferably in an amount from 50-99% by weight of the composition, more preferably in an amount from 75-99% by weight of the composition, even more preferably in an amount from 80-97% by weight of the composition, most preferably in an amount from 85-97% by weight of the composition. The microcrystalline cellulose may be the only or sole filler present in the composition, i.e. the composition may be free from other fillers than microcrystalline cellulose. In another embodiment, the composition comprises lactose monohydrate. The lactose monohydrate is typically present in an amount from 10-99% by weight of the composition, preferably in an amount from 50-99% by weight of the composition, from 75-99% by weight of the composition, from 80-97% by weight of the composition, or from 85-97% by weight of the composition. The lactose monohydrate may be the only or sole filler present in the composition, i.e. the composition may be free from other fillers than lactose monohydrate. In some embodiments, the composition described herein includes microcrystalline cellulose and lactose monohydrate. The microcrystalline cellulose is typically present in an amount from 20-80% by weight of the composition and lactose monohydrate in an amount from 20-80% byweight of the composition. In one embodiment, microcrystalline cellulose constitutes the major part of the microcrystalline cellulose-lactose monohydrate filler system, i.e. the composition comprises lactose monohydrate in an amount from 20-60% by weight of the composition and microcrystalline cellulose in an amount from 40-80% by weight of the composition, such as lactose monohydrate in an amount from 20-45% by weight of the composition and microcrystalline cellulose in an amount from 40-70% by weight of the composition, e.g. lactose monohydrate in an amount from 25-36% by weight of the composition and microcrystalline cellulose in an amount from 52-63% by weight of the composition. The microcrystalline cellulose and the lactose monohydrate may be the only fillers present in the composition, i.e. the composition may be free from other fillers than microcrystalline cellulose and lactose monohydrate. In another embodiment, lactose monohydrate constitutes the major part of the microcrystalline cellulose-lactose monohydrate filler system, i.e. the composition comprises microcrystalline cellulose in an amount from 20-60% by weight of the composition and lactose monohydrate in an amount from 40-80% by weight of the composition. In some embodiments, the composition described herein includes microcrystalline cellulose in an amount from 20-45% by weight of the composition and lactose monohydrate in an amount from 40-70% by weight of the composition. In some embodiments, the composition described herein includes microcrystalline cellulose in an amount from 25-36% by weight of the composition and lactose monohydrate in an amount from 52-63% by weight of the composition. The microcrystalline cellulose and the lactose monohydrate may be the only fillers present in the composition, i.e. the composition may be free from other fillers than microcrystalline cellulose and lactose monohydrate.

[0086] Microcrystalline cellulose is commercially available in different particle sizes and moisture grades. Examples of commercially available microcrystalline cellulose preparations include the Avicel® PH-series from FMC Biopolymer, the Emcocel® M-series from Penwest Pharmaceuticals Co. and the Vivapur®-series from Rettenmaier & Söhne GmbH. A parti cul ar preferred commercial product to be used for the purposes described herein is Avicel® PH-101. Likewise, various lactose monohydrate grades having different physical properties, such as particle size distribution and flow characteristics, are commercially available. The grade of the lactose monohydrate may vary dependent on the specific dosage form to be prepared. For example, direct-compression grades of lactose monohydrate, such asTablettose® (agglomerated) or grades for powder blends, such as Pharmatose® DCL 11 (spray-dried), have better flow properties and are more compressible than powdered or crystalline lactose monohydrate. Such lactose monohydrate preparations are not particularly preferred for the purposes described herein. Rather, the more fine grade lactose monohydrate preparations are preferred, such as powdered or crystalline lactose monohydrate, in particular crystalline lactose monohydrate where 90% of the particles have a diameter of less than 0.1 mm.

[0087] In some embodiments, the composition described herein includes one or more binders. Examples of a binder include but are not limited to sucrose, glucose, sorbitol, acacia, alginic acid, sodium alginate, gelatine, starch, pregelatinised starch, magnesium aluminium silicate, carboxymethylcellulose sodium (CMC sodium), methylcellulose, ethylcellulose, hydroxypropylmethylcellulose (HPMC), hydroxy-propylcellulose (HPC), polyvinylacetate or polyethylene glycol. The binder is typically present in an amount from 0.1-10% by weight of the composition. In some embodiments, the binder is present in an amount from 0.2-5% by weight of the composition, such as from 0.5-5% by weight of the composition or from 1-3% by weight of the composition. In some embodiments, the binder is HPC or polyvinylpyrrolidone (PVP). In some embodiments, the binder is polyvinyl alcohol (PVA).

[0088] In some embodiments, the composition described herein includes one or more lubricants, including glidants and antiadhesives, such as magnesium stearate, zinc stearate, stearic acid, silicas, hydrogenated vegetable oils or talc. The lubricant is typically present in an amount from 0.1-10% by weight of the composition. In some embodiments, the lubricant is present in an amount from 0.2-5% by weight of the composition, such as from 0.5- 5% by weight of the composition or from 1-3% by weight of the composition. In some embodiments, the lubricant is magnesium stearate.

[0089] In some embodiments, the composition described herein includes one or more disintegrants. Examples of the disintegrants include but are not limited to sodium starch glycol ate, maize starch, rice starch, potato starch, cross-linked povidone or carboxymethylcellulose-based disintegrants. Carboxymethylcellulose-based disintegrants may be present as free acid, but is preferably in the form of a salt, e.g. in the form of an alkali metal salt, such as the potassium salt or the sodium salt, in particular the sodium salt, or in the form of a salt of a divalent metal ion, such as the magnesium salt, the calcium salt or the zinc salt,in particular the calcium salt. The carboxymethylcellulose-based disintegrant may be cross-linked or non-cross-linked. Specific examples of preferred non-cross-linked carboxymethylcellulose-based disintegrants include carboxymethyl-cellulose calcium (carmellose calcium) and carboxymethylcellulose sodium (carmellose sodium), in particular carboxymethylcellulose calcium. In some embodiments, the carboxymethylcellulose-based disintegrant is cross-linked. A specific example of a preferred cross-linked carboxymethylcellulose-based disintegrant is cross-linked carboxymethylcellulose sodium (croscarmellose sodium). Croscarmellose sodium is commercially available under the tradenames Ac-Di-Sol®, Explocel® and Solutab®. The disintegrant is typically present in an amount from 0.1-10% by weight of the composition. Preferably, the disintegrant is present in an amount from 0.2-5% by weight of the composition, such as from 0.5-5% by weight of the composition, more preferably in an amount from 1-4% by weight of the composition.

[0090] In some embodiments, the composition described herein includes one or more surfactants and wetting agents. Examples of the surfactants and wetting agents include but are not limited to naturally occurring phosphatides, e.g. lechitin or soybean lechitin; condensation products of ethylene oxide with e.g. a fatty acid, a long chain fatty alcohol, or a partial ester derived from fatty acids and a hexitol or a hexitol anhydride, for example polyoxyethylene stearate, polyoxyethylene sorbitol monooleate, polyoxyethylene sorbitan monooleate, etc.; or salts of long-chain aliphatic phosphates, such as sodium lauryl sulphate.

[0091] In those cases where the pharmaceutical formulation is in the form of a solid oral dosage form, in particular a solid unit dosage form (e.g. a tablet, sachet or capsule, in particular a tablet), the dosage form is adapted for oral administration and may be provided with a coating, such as a film coating, a sugar coating, or the like. Thus, a suitable coating for the dosage form may, for example, be a sugar coating or a film coating based on one or more of the ingredients: Hydroxypropylmethylcellulose (HPMC), methyl cellulose, ethylcellulose, hydroxyethylmethylcellulose, hydroxypropylcellulose, carboxymethylcellulose sodium, acrylate polymers (e.g. Eudragit®), polyethylene glycols or polyvinylpyrrolidone.

[0092] In some embodiments, the composition is in the form of a tablet, a coated tablet, or a film-coated tablet. In some embodiments, the uncoated tablet typically has a weight in the range from 10-600 mg, 50-150 mg, 60-125 mg, 60-100 mg, or 70-90 mg.

[0093] In some embodiments, the composition described herein is prepared by a freeze-dry process. In some embodiments, the freeze-dry process includes combining cyclodextrin and folic acid or folic acid derivative in a solution at a ratio that is sufficient to reduce or prevent degradation of the folic acid or folic acid derivative and subject the solution to a freeze-dry process. In some embodiments, the composition described herein is prepared by a spray-dry process. In some embodiments, the spray-dry process includes combining cyclodextrin and folic acid or folic acid derivative in a solution at a ratio that is sufficient to reduce or prevent degradation of the folic acid or folic acid derivative and subject the solution to a spray-dry process.

[0094] In some embodiments, the compositions described herein may be prepared for intravenous or intramuscular administration via injection. In some such embodiments, the compositions described herein may be dissolved or dispersed in a pharmaceutically acceptable diluent, such as a saline or dextrose solution. Suitable excipients may be included to achieve the desired pH, including but not limited to NaOH, sodium carbonate, sodium acetate, HCl, and citric acid. Accordingly, buffers including acetate buffers, citrate buffers, phosphate buffers and borate buffers may be used to adjust the pH of these formulations as needed. Antioxidant excipients may include sodium bisulfite, acetone sodium bisulfite, sodium formaldehyde, sulfoxylate, thiourea, and edetate disodium. Other non-limiting examples of suitable excipients found in the final intravenous composition may include sodium or potassium phosphates, citric acid, tartaric acid, gelatin, and carbohydrates such as dextrose, mannitol, and dextran. Further acceptable excipients are described in Powell, et al., Compendium of Excipients for Parenteral Formulations, PDA J P harm Sci and Tech 1998, 52 238-311 and Nema et al., Excipients and Their Role in Approved Injectable Products: Current Usage and Future Directions, PDA J Pharm Sci and Tech 2011, 65287-332, both of which are incorporated herein by reference in their entirety. Antimicrobial agents may also be included to achieve a bacteriostatic or fungistatic solution, including but not limited to phenylmercuric nitrate, thimerosal, benzethonium chloride, benzalkonium chloride, phenol, cresol, and chlorobutanol.

[0095] The compositions for intravenous or intramuscular administration may be provided to caregivers in the form of one more solids that are reconstituted with a suitable diluent such as sterile water, saline or dextrose in water shortly prior to administration. Inother embodiments, the compositions are provided in solution ready to administer parenterally. In still other embodiments, the compositions are provided in a solution that is further diluted prior to administration. In embodiments that include administering a combination of a composition described herein and another agent, the combination may be provided to caregivers as a mixture, or the caregivers may mix the two agents prior to administration, or the two agents may be administered separately.Liquid Oral Formulations

[0096] Some embodiments provide an oral formulation comprising a compound according to Formula (II) and a cyclodextrin. In some embodiments, the oral formulation is a solution. In other embodiments, the formulation is prepared from a powder reconstituted into a solution. In some embodiments, the solvent is water. The oral formulation comprises a compound according to Formula (II) that may be present at a concentration of 0.01 mg / mL, 0.1 mg / mL, 1 mg / mL, 2 mg / mL, 5 mg / mL, 10 mg / mL, 25 mg / mL, 50 mg / mL, 100 mg / mL, 250 mg / mL, 500 mg / mL, 1,000 mg / mL, or a concentration in between any of the aforementioned concentrations. In some embodiments, the compound is Levo Leucovorin or pharmaceutically acceptable salts thereof. In some embodiments, the cyclodextrin has the structure according to Formula (I). In some embodiments, the cyclodextrin is sulfobutylether-P-cyclodextrin.

[0097] In some embodiments, the amount of liquid oral formulation administered per dose is 0.1 mL, 0.5 mL, 1 mL, 2.5 mL, 5 mL, 7.5 mL, 10 mL, 15 mL, 20 mL, 25 mL, 50 mL, 100 mL, or an amount in between any of the aforementioned volumes.

[0098] In some embodiments, the dose of Levo Leucovorin or pharmaceutically acceptable salt thereof is 0.01 mg, 0.1 mg, 1 mg, 2 mg, 3mg, 5 mg, 10 mg, 15 mg, 25 mg, 50 mg, 75 mg, 100 mg, 175 mg, 200 mg, 250 mg, 350 mg, 500 mg, 1,000 mg, or an amount in between any of the aforementioned values.

[0099] In some embodiments, the present disclosure provides liquid pharmaceutical compositions comprising Levo Leucovorin or a derivative thereof, a cyclodextrin according to Formula (I) (e.g., sulfobutyl ether- -cyclodextrin), and optionally one or more pharmaceutically acceptable excipients.

[0100] In some embodiments, the formulation is room temperature stable. In some embodiments, the pharmaceutical composition is a liquid composition that requires no reconstitution. In some embodiments, the pharmaceutical compositions may not require shaking or mixing prior to use.

[0101] In some embodiments, the liquid pharmaceutical compositions may be in the form of a liquid dispersion, a suspension, a solution, an emulsion, a spray, a syrup, an elixir, a drop, a concentrate, or a combination thereof. In some embodiments, the pharmaceutical composition may be in the form of a dry powder to be prepared into a liquid pharmaceutical composition prior to administration to a patient.

[0102] In some embodiments, the liquid pharmaceutical compositions of the present disclosure may also comprise anti -microbial agents or preserving agents or preservatives. In some embodiments, the amount of anti-microbial agent, preserving agent, or preservative may be less than without a cyclodextrin according to Formula (I). In some embodiments, the composition does not comprise a preservative.

[0103] In some embodiments, the present disclosure therefore provides palatable liquid compositions comprising Levo Leucovorin and at least one or both selected from sweeteners / sweetening agents and flavoring agents.

[0104] The liquid compositions according to the present disclosure, without limitation include, aqueous dosage forms, alcoholic and / or hydro-alcoholic dosage forms and non-aqueous dosage forms. Aqueous dosage forms according to the present disclosure may also comprise one or more non-aqueous and / or organic solvents.

[0105] In some of the embodiments, the present disclosure provides the liquid pharmaceutical compositions in the form of spray which may be administered by oral route. Sprays are known by various names such as aerosol sprays, liquid pump sprays, or activated mists etc.

[0106] In some of the embodiments, the liquid pharmaceutical compositions of the present disclosure are in the form of immediate release dosage forms or modified release dosage forms, such as extended release, controlled release, sustained release, prolonged release and delayed release. In some of the embodiments, the liquid pharmaceutical compositions comprise Levo Leucovorin and one or more suitable excipients or additives for the preparation of modified release dosage forms such as rate controlling polymers.

[0107] The liquid pharmaceutical compositions of the present disclosure may also be prepared by reconstitution of dry powder in suitable diluent or media such as water. The dry powder for reconstitution may be in the form of immediate release forms. The dry powder composition may comprise a compound of Formula (II) (e g., Levo Leucovorin), a cyclodextrin, and one or more suitable excipients selected from the group comprising of fillers, binders, diluents, disintegrants, pore formers, lubricants, glidants, sweeteners, stabilizing agents, antioxidants, flavoring agents, viscosity modifying agents, surfactants, preservatives and plasticizers. The dry powder for reconstitution may also be in the form of modified release forms and comprise one or more suitable excipients such as rate controlling polymers.

[0108] Some embodiments include a method of administering a liquid oral formulation comprising providing a dry powder comprising a compound of Formula (II) (e.g., Levo Leucovorin) and a cyclodextrin (e.g., sulfobutylether-P-cyclodextrin), adding a diluent (e.g., water) to the dry powder, mixing until a uniform solution or suspension is achieved, and then administering the solution or suspension.

[0109] In some embodiments, the liquid oral formulation, or dry powder for reconstitution, further comprises a buffering agent. In some embodiments, the buffering agent is in an amount sufficient to make the pH of the pharmaceutical composition about 6.5 to about 8.5. In some embodiments, the buffer is selected from the group comprising Tris, ammonium acetate, citrate, phosphate, MES, and HEPES.

[0110] In some embodiments, the pH of the compositions is between about 2.0 and about 11.0. In some of the embodiments, the pH of the compositions is between about 4.0 and about 9.0. In some of the embodiments, the pH of the compositions is between about 4.5 and about 8.5. In some of the embodiments, the pH of the compositions is between about 7.0 and about 8.0. In some of the embodiments, the pH of the compositions is between about 7.5 and about 7.9. In some of the embodiments, the pH of the composition is about 7.8.

[0111] In some embodiments, the liquid pharmaceutical compositions of the present disclosure are stable for prolonged time when stored under storage conditions. The term “storage conditions” as used herein without limitation include typical storage conditions such as 2° C.-8° C., 40° C.±2° C. / 75±5% RH, 30° C.±2° C. / 65±5% RH, 25° C.±2° C. / 40±5% RH, 25° C.±2° C. / 60±5% RH, and 40° C ±2° C. / NMT 25% RH (NMT=not more than) and accelerated conditions such as 40° C.±2° C. / 75±5% RH. The term “prolonged time” as usedherein indicates that the liquid pharmaceutical compositions of the present disclosure are stable for at least 1 month, at least 3 months, at least 6 months or at least 12 months when stored under storage conditions.

[0112] In some of the embodiments of the present disclosure, stable liquid pharmaceutical compositions or stability of the liquid pharmaceutical compositions refer to compositions which retain at least about 90%, or about least about 95%, or at least about 96%, or at least about 98%, of the labelled concentration of compound of Formula (II) (e.g., Levo Leucovorin) contained in the said composition after storage under typical and / or accelerated conditions. In further embodiments, stable liquid pharmaceutical compositions or stability of the liquid pharmaceutical compositions refer to less than about 15% (area percent), or less than about 10% (area percent), or less than about 7% (area percent), or less than about 5% (area percent), or less than about 2% (area percent) of impurities are present after storage under typical and / or accelerated conditions.

[0113] Methods for determining the stability of the liquid pharmaceutical compositions of the present disclosure with respect to a given parameter are well-known to those of skill in the art. For example, individual impurities and total impurities can be assessed by high-performance liquid chromatography (HPLC) or thin layer chromatography (TLC). Unless otherwise indicated to the contrary, a percentage amount of any individual impurities (known / unknown), or total impurities reported herein in the liquid compositions are determined by a peak area percent method using HPLC.

[0114] In some of the further embodiments, the present disclosure provides concentrate liquid compositions which may be diluted using suitable diluent before administering to the patient. In some of the embodiments, the liquid compositions of the invention are ready-to-use liquid compositions. Such ready-to-use compositions of the invention are administered directly to the patients in required doses without any prior preparation e.g. reconstitution in suitable diluent such as water.

[0115] Following embodiments of the invention describe suitable excipients which may be used to prepare liquid compositions of the present disclosure. It is in no way the intention of the present inventor(s) / applicant(s) to limit the scope of the liquid compositions of the present disclosure by the description of following embodiments. Described embodiments are for illustrative purpose only and a skilled person may use other excipients from the sameor different classes as well which may provide liquid compositions of the present disclosure same or improved physico-chemical properties, palatability, stability and the like and retain or increase patients' acceptability towards the therapy. Such other excipients, classes of excipients and compositions resulted therefrom are also part of the present disclosure and covered within the scope of the present disclosure.

[0116] Vehicles may be used in the liquid compositions of the present disclosure. Vehicles are the liquid bases that carry drugs and other excipients in dissolved or dispersed state. Vehicles may be aqueous or non-aqueous or mixture thereof. In some embodiments, the vehicle is OraSweet, OraSweet SF, or Orabase. Non-aqueous solvents / cosolvents may also be added in the liquid compositions of the present disclosure. Suitable solvents / co-solvents, solubilizers or vehicles, that may be employed, in the liquid compositions of embodiments herein include, but are not limited to, dichloromethane, acetonitrile, ethyl acetate, acetone, propylene carbonate, water, glycerin, coconut fatty acid di ethanol ami de, medium and / or long chain fatty acids or glycerides, monoglycerides, diglycerides, triglycerides, structured triglycerides, soyabean oil, peanut oil, com oil, corn oil monoglycerides, corn oil diglycerides, corn oil triglycerides, polyethylene glycol, caprylocaproylmacroglycerides, caproyl 90, propylene glycol, polyoxyethylenesorbitan fatty acid esters, polyoxyethylene castor oil derivatives, castor oil, cottonseed oil, olive oil, safflower oil, peppermint oil, coconut oil, palm seed oil, beeswax, oleic acid, methanol, ethanol, isopropyl alcohol, butanol, acetone, methyl isobutyl ketone, methyl ethyl ketone and the like or any combinations thereof.

[0117] Non-limiting examples of wetting agents are Benzalkonium chloride, Benzethonium chloride, Cetylpyridinium chloride, Docusate sodium, Nonoxynol 9, Octoxynol, Poloxamer, Poloxamer 124, Poloxamer 188, 237, 338, 407, Polyoxyl 35 castor oil, Polyoxyl 40 hydrogenated castor oil, Polyoxyl 10 oleyl ether, Polyoxyl 20 cetylstearyl ether, Polyoxyl 40 stearate, Polysorbate 20, Polysorbate 40, Polysorbate 60, Polysorbate 80, Sodium lauryl sulfate, Sorbitan monolaurate, Sorbitan monooleate, Sorbitan monopalmitate, Sorbitan monostearate, Tyloxapol and the like or any combinations thereof.

[0118] The amount of wetting agent, e.g. glycerin, is generally about 200 mg / mL to about 400 mg / mL. In some embodiments, the wetting agent is in an amount of about 100 mg / mL to about 1000 mg / mL, about 100 mg / mL to about 800 mg / mL, about 100 mg / mL to about 600 mg / mL, about 100 mg / mL to about 500 mg / mL, about 100 mg / mL to about 400mg / mL, about 200 mg / mL to about 1000 mg / mL, about 200 mg / mL to about 800 mg / mL, about 200 mg / mL to about 600 mg / mL, about 200 mg / mL to about 500 mg / mL, or a value between any of these ranges.

[0119] Solubility enhancing agents may include, but are not limited to, DL-methionine, caffeine, nicotinamide, vanillin, benzyl alcohol, ethanol and diethylene glycol monoethyl ether and the like or combinations thereof.

[0120] Stabilizing agents may include, but are not limited to, sodium metabisulphite, sodium bisulphite, ethylene diamine tetraacetic acid (EDTA) or salts thereof, ascorbic acid and the like or combinations thereof.

[0121] Penetration / permeation enhancers may include, but are not limited to, nicotinamide, caffeine, peppermint oil, sodium glycocholate, phospholipids, alkyl saccharides, aprotinin, benzalkonium chloride, ceramides, cetylpyridiniuni chloride, chitosan, chitosan-4-thiobutylamidine, cyclodextrins, dextran sulfate, dodecyl azacycloheptyl-2-ketone, ether lipids (plasm ologens), glycerol, glycosylated sphingosines, lauric acid, 23 -lauryl ether, lysophosphatidyl choline, menthol, meth oxy salicylate, phosphatidyl choline, l-palmitoyl-2-glutaroyl-sn-glycero-3-phosphocholine, polycarbophil cysteine, poly-L-arginine, polyoxyethylene, polyoxyethylene-9-lauryl ether, polysorbate 80, propylene glycol, EDTA, sodium deoxycholate, sodium glycocholate, sodium glycodeoxy cholate, sodium lauryl sulfate, sodium salicylate, sodium taurocholate, sodium taurodeoxycholate, sodium taurodihydrofusidate, sphingolipids, sterols and the like or combinations thereof.

[0122] Mucoadhesives may also be added in the compositions of the present disclosure. Examples of suitable mucoadhesives include, but are not limited to, hydroxypropyl cellulose, gelatin, crosslinked polyacrylic acid, polymethacrylic acid, polyhydroxyethyl methacrylic acid, hydroxypropyl methyl cellulose, polyethylene glycol, sodium carboxymethyl cellulose, hyaluronic acid, chitosan, polycarbophil, pectin, xanthan gum, alginate, copolymers of dextran, polyacrylamide, acacia, copolymer of caprolactone and ethylene oxide, carbopol 934, tragacanth, eudragit and the like or combinations thereof

[0123] Viscosity modifying agents, also referred to herein as suspending agents or thickening agents, impart viscosity, and thus retard particle sedimentation. Other factors considered in the selection of the appropriate agent include desired rheological property, suspending ability in the system, chemical compatibility with other excipients, pH stability,length of time to hydrate, batch-to-batch reproducibility, and cost. Non-limiting examples of pH adjusting agents / modifiers and buffers are Acetic acid, Adipic acid, Ammonium carbonate, Ammonium hydroxide, Ammonium phosphate, Boric acid, Citric acid, Diethanolamine, Fumaric acid, Hydrochloric acid, Malic acid, Nitric acid, Propionic acid, Potassium acetate, Potassium bicarbonate, Potassium chloride, Potassium citrate, Potassium metaphosphate, Potassium phosphate, Sodium acetate, Sodium bicarbonate, Sodium borate, Sodium carbonate, Sodium chloride, Sodium citrate, Sodium glycolate, Sodium hydroxide, Sodium lactate, Sodium phosphate, Sodium proprionate, Succinic acid, Sulfuric acid, Tartaric acid, Tri ethylamine, Triethanolamine, Tromethamine, Trolamine and the like or any combinations thereof

[0124] Viscosity modifying agents can be classified into cellulose derivatives, clays, natural gums, and synthetic gums. In many cases, these excipients are used in combination. There are many water soluble hydrocolloids that can act as viscosity modifying agents in the formulation of pharmaceutical suspensions. They can be of natural, semi¬ synthetic or synthetic origin. Non-limiting examples of viscosity modifying agents are Acacia, Agar, Alginic acid, Carbomer, Carmellose sodium, Dextrin, Gelatin, Veegum or Gel white, Gellan gum, Sodium alginate, Methylcellulose, Hydroxyethyl cellulose, Hydroxypropyl cellulose, Hydroxypropylmethyl cellulose. Hydroxypropyl starch, Hypromellose, Maltodextrin, Methylcellulose, Modified starch, Pectin, Poloxamer, Polycarbophil, Polyethylene glycol, Polyvinyl acetate, Poly (vinyl alcohol), Potassium alginate, Polyvinyl pyrrolidone, Pregelatinized starch, Propylene glycol alginate, Sodium alginate, Carboxymethyl cellulose or an alkali metal salt thereof, Microcrystalline cellulose, gum Arabic, Karaya gum, Sterculia gum, Tragacanth, Xanthan gum. Bentonite, Carageenan, Guar gum, Colloidal silicon dioxide and the like or any combinations thereof.

[0125] In some embodiments, the viscosity modifying agent is present in an amount of about 1 mg / mL to about 10 mg / mL, about 1 mg / mL to about 20 mg / mL, about 1 mg / mL to about 15 mg / mL, about 2 mg / mL to about 20 mg / mL, about 2 mg / mL to about 15 mg / mL, about 2 mg / mL to about 10 mg / mL, about 2 mg / mL to about 8 mg / mL, about 2 mg / mL to about 6 mg / mL, about 4 mg / mL to about 20 mg / mL, about 4 mg / mL to about 15 mg / mL, about 4 mg / mL to about 10 mg / mL, about 4 mg / mL to about 8 mg / mL, about 6 mg / mL toabout 20 mg / mL, about 6 mg / mL to about 15 mg / mL, about 8 mg / mL to about 20 mg / mL, about 8 mg / mL to about 15 mg / mL, and any value within the foregoing ranges.

[0126] Non-limiting examples of preservatives are Alcohol, Ethanol, Chlorobutanol, Phenoxyethanol, Potassium benzoate, Benzyl alcohol, Benzoic acid, Potassium sorbate, Sorbic acid, Benzalkonium chloride, Benzethonium chloride, Cetrimonium bromide, Cetylpyridinium chloride, Bronopol, Chlorbutol, Chlorocresol, Cresol, Butylparaben, Methylparaben, Propylparaben, Ethylparaben, Phenol, Thymol, Phenyl ethanol, Sodium benzoate, Antimicrobial solvents like Propylene glycol, Glycerin, Chloroform and the like or any combinations thereof. In addition, some formulation ingredients like nonionic surfactants, quaternary ammonium compounds, gelatin, ferric salts, calcium salts and salts of heavy metals, including silver, lead, and mercury prevent microbial growth.

[0127] In some embodiments, the preservative is present in an amount of about 0.1 mg / mL to about 10 mg / mL, about 0.1 mg / mL to about 20 mg / mL, about 0.1 mg / mL to about 15 mg / mL, about 2 mg / mL to about 20 mg / mL, about 2 mg / mL to about 15 mg / mL, about 2 mg / mL to about 10 mg / mL, about 2 mg / mL to about 8 mg / mL, about 2 mg / mL to about 6 mg / mL, about 4 mg / mL to about 20 mg / mL, about 4 mg / mL to about 15 mg / mL, about 4 mg / mL to about 10 mg / mL, about 4 mg / mL to about 8 mg / mL, about 6 mg / mL to about 20 mg / mL, about 6 mg / mL to about 15 mg / mL, about 8 mg / mL to about 20 mg / mL, about 8 mg / mL to about 15 mg / mL, and any value within the foregoing ranges.

[0128] Non-limiting examples of antioxidants are a-Tocopherol acetate, Ascorbic acid, Erythorbic acid, Butylated hydroxytoluene (BHT), d-a-Tocopherol natural, Monothioglycerol, Sodium bisulfite, Sodium sulfite, Sodium metabisulfite, Potassium metabisulfite, Acetone sodium bisulfite, Ascorbyl palmitate, Cysteine, d-a-tocopherol synthetic, Nordihydroguaiaretic acid, Sodium formaldehyde sulfoxylate, Sodium thiosulfate, Acetylcysteine, Ascorbyl palmitate, Butylated hydroxy anisole (BHA), Cysteine hydrochloride, Dithiothreitol, Propyl gallate, Thiourea and the like or any combinations thereof.

[0129] Non-limiting examples of bulking agents are Calcium carbonate, Calcium hydroxide, Cellulose, Crospovidone, Dibasic calcium phosphate, Magnesium carbonate, Magnesium hydroxide, Microcrystalline cellulose. Silica (silicon dioxide), Titanium dioxide and the like or any combinations thereof.

[0130] Non-limiting examples of surfactants are Sodium lauryl sulfate, Docusate sodium, Cocamidopropyl amino betaine, Polyoxyethylene sorbitan fatty acid esters (Polysorbate, Tween®), Polyoxyethylene 15 hydroxystearate (Macrogol 15 hydroxystearate, Solutol HS15®), Polyoxyethylene castor oil derivatives (Cremophor® EL, ELP, RH 40), Polyoxyethylene stearates (Myrj®), Sorbitan fatty acid esters (Span®), Polyoxyethylene alkyl ethers (Brij®), Polyoxyethylene nonylphenol ether (Nonoxynol®) and the like or any combinations thereof. Anti-foaming agents may be used in the preparation of the liquid pharmaceutical compositions of the present disclosure to lower the surface tension and cohesive binding of liquid phase. Non-limiting examples of anti-foaming agents are simethicone, organic phosphates, alcohols, paraffin oils, stearates, glycols and the like or any combinations thereof. In some embodiments, the anti-foaming agent is 30% simethicone emulsion.

[0131] Non-limiting examples of chelating agents are Calcium disodium edetate, Disodium edetate, Edetic acid (also known as ethylenediaminetetraacetic acid / EDTA), Citric acid and the like or any combinations thereof.

[0132] Non-limiting examples of sweetening agents are Glucose, Sucralose, Trehalose, Fructose, Xylose, Dextrose, Galactose, Tagatose, Maltose, Sucrose, Glycerol, Dulcitol, Mannitol, Lactitol, Sorbitol, Xylitol, Saccharine or the corresponding sodium, potassium or calcium salt, Cyclamate or the corresponding sodium or calcium salt, Aspartame, or Acesulfame or the potassium salt thereof, Dulcin or Ammonium glycyrrhizinate, Alitame, Inulin, Isomalt, Neohesperidin dihydrochalcone, Thaumatin and the like or any combinations thereof.

[0133] In some embodiments, the sweetening agent is present in an amount of about 1 mg / mL to about 10 mg / mL, about 1 mg / mL to about 20 mg / mL, about 1 mg / mL to about 15 mg / mL, about 2 mg / mL to about 20 mg / mL, about 2 mg / mL to about 15 mg / mL, about 2 mg / mL to about 10 mg / mL, about 2 mg / mL to about 8 mg / mL, about 2 mg / mL to about 6 mg / mL, about 4 mg / mL to about 20 mg / mL, about 4 mg / mL to about 15 mg / mL, about 4 mg / mL to about 10 mg / mL, about 4 mg / mL to about 8 mg / mL, about 6 mg / mL to about 20 mg / mL, about 6 mg / mL to about 15 mg / mL, about 8 mg / mL to about 20 mg / mL, about 8 mg / mL to about 15 mg / mL, about 25 mg / mL, about 50 mg / mL, about 100 mg / mL, about 150mg / mL, about 200 mg / mL, about 250 mg / mL, about 500 mg / mL and any value within the foregoing ranges.

[0134] Non-limiting examples of flavoring agents are synthetic flavor oils and flavoring aromatics and / or natural oils, extracts from plants leaves, flowers, fruits, and so forth and the like or any combinations thereof. These may include cinnamon oil, oil of wintergreen, peppermint oils, clove oil, bay oil, anise oil, eucalyptus, thyme oil, cedar leaf oil, oil of nutmeg, oil of sage, oil of bitter almonds, and cassia oil and the like or any combinations thereof. Also useful as flavors are vanilla, citrus oil, including lemon, orange, grape, lime and grapefruit, and fruit essences, including apple, banana, pear, peach, berry, strawberry, raspberry, cherry, plum, pineapple, apricot, bubblegum, tutti fruiti, and so forth and the like or any combinations thereof. Solid forms, such as spray dried forms of flavoring agents, may also be useful in the liquid compositions disclosed herein.

[0135] In some embodiments, the flavoring agent is present in an amount of about 0.5 mg / mL to about 10 mg / mL, about 1 mg / mL to about 20 mg / mL, about 1 mg / mL to about 15 mg / mL, about 2 mg / mL to about 20 mg / mL, about 2 mg / mL to about 15 mg / mL, about 2 mg / mL to about 10 mg / mL, about 2 mg / mL to about 8 mg / mL, about 2 mg / mL to about 6 mg / mL, about 4 mg / mL to about 20 mg / mL, about 4 mg / mL to about 15 mg / mL, about 4 mg / mL to about 10 mg / mL, about 4 mg / mL to about 8 mg / mL, about 6 mg / mL to about 20 mg / mL, about 6 mg / mL to about 15 mg / mL, about 8 mg / mL to about 20 mg / mL, about 8 mg / mL to about 15 mg / mL, and any value within the foregoing ranges.Methods of Maintaining Folic Acid or Folic Acid Derivative Stability

[0136] Some embodiments relate to a method of reducing an oxidative degradation of a folic acid or folic acid derivative, comprising combining an amount of folic acid or folic acid derivative and an amount of cyclodextrin, wherein the amount of cyclodextrin is effective in reducing the oxidative degradation of the folic acid or folic acid derivative.

[0137] In some embodiments, the molar ratio of the cyclodextrin to the folic acid or folic acid derivative is in the range of about 0.1 to 10, about 0.1 to 8, about 0.1 to 5, about 0.5 to 2, about 0.5 to 1.5, or about 0.5 to 1.25. In some embodiments, the molar ratio of the cyclodextrin to the folic acid or folic acid derivative is greater than 0,05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.05, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.5, 3, 4, 5, or 6. In someembodiments, the molar ratio of the cyclodextrin to the folic acid or folic acid derivative is less than 1, 1.05, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.5, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, the molar ratio of the cyclodextrin to the folic acid or folic acid derivative is about 1.0 to about 1.2. In some embodiments, the molar ratio of the cyclodextrin to the folic acid or folic acid derivative is about 1.1.

[0138] In some embodiments, the weight ratio of the cyclodextrin to the folic acid or folic acid derivative is in the range of about 0.1 to 20, about 1 to 10, about 1 to 8, about 2 to 8, about 3 to 7, or about 4 to 6. In some embodiments, the weight ratio of the cyclodextrin to the folic acid or folic acid derivative is greater than 0.1, 0.5, 1, 2, 3, 3.5, 4, 4.5, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.7, 5.9, 6, 7, 8, 9, or 10. In some embodiments, the weight ratio of the cyclodextrin to the folic acid or folic acid derivative is less than 3, 4, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.7, 5.9, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. In some embodiments, the weight ratio of the cyclodextrin to the folic acid or folic acid derivative is about 4.8 to about 5.3. In some embodiments, weight ratio of the cyclodextrin to the folic acid or folic acid derivative is about 5.1.

[0139] In some embodiments, the cyclodextrin and folic acid or folic acid derivative are combined in a solution at a ratio described herein prior to being freeze-dried. In some embodiments, the freeze-dried composition can be forced through a 40-mesh screen.

[0140] In some embodiments, the cyclodextrin and folic acid or folic acid derivative are combined in a solution at a ratio described herein prior to being spray-dried.Method s of Treatment

[0141] Some embodiments relate to a method of preventing birth defects in a subject comprising administering to the subject a composition described herein. In some embodiments, the birth defect may be a neural tube defect, a cleft lip defect, or a cleft palate defect.

[0142] Some embodiments relate to a method of treating a deficiency or disease positively affected by the administration of both folates and reduced folates selected from the group consisting of subacute encephalitis associated with dementia, vacuolar myelopathy, premature occlusive arterial disease, severe vascular disease in infancy, severe vascular disease in childhood, progressive arterial stenosis, intermittent claudication, renovascularhypertension, ischemic cerebrovascular disease, premature retinal artery occlusion, premature retinal vein occlusion, cerebral occlusive arterial disease, occlusive peripheral arterial disease, premature death due to thromboembolic disease, premature death due to ischemic heart disease, psoriasis, celiac disease, arthritic condition, inflammation conditions, megaloblastic anemia due to folate deficiency, intestinal malabsorption, and depressive illness, said method comprising administering to a subject in need thereof an effective amount of the composition described herein.

[0143] Some embodiments relate to a method of reducing side effects of a drug or active agent, said method comprising administering to a subject in need thereof an effective amount of any one of the compositions described herein.

[0144] Some embodiments relate to a method of treating cancer, said method comprising administering to a subject in need thereof an effective amount of any one of the compositions described herein. In some embodiments, the cancer is colorectal cancer, pancreatic cancer, gallbladder and biliary tree carcinoma, gastric cancer, squamous cell carcinoma of the head and neck, cervical cancer, esophageal cancer, skin cancer, bladder cancer, Kaposi’s sarcoma, acute lymphocytic leukemia, lymphoma, lung cancer, and uterine cancer. In some embodiments, the method may further comprise the administration of an additional active agent. In some embodiments, the additional active agents and composition described herein may be administered sequentially. In other embodiments, the additional active agents and composition described herein may be administered simultaneously. In some embodiments, the additional active agent may be a chemotherapeutic agent. In some embodiments, the additional active agent may be a folic acid antagonist. In some embodiments, the additional active agent may be methotrexate, pyrimethamine, pralatrexate, 5-fluorouracil, doxorubicin, epirubicin, idarubicin, daunorubicin, and cyclophosphamide

[0145] Some embodiments relate to a method for hormone replacement therapy in a subject comprising administering to the subject a composition described herein.

[0146] Some embodiments relate to a method of reducing a fem ale's risk of having a miscarriage, reducing a female's risk of having a fetus with a neural tube defect, reducing a femal e’s risk of having a fetus with a cleft lip defect, reducing a female's risk of having a fetus with a cleft palate defect, said method comprising administering to a subject in need thereof an effective amount of any one of the composition described herein.

[0147] Some embodiments provided herein relate to a method of treating or preventing a neurodevel opmental disorder, said method comprising administering to a subject in need thereof an effective amount of any one of the compositions described herein. In some embodiments, the neurodevel opmental disorder may be fragile X syndrome, autism spectrum disorder, attention deficit hyperactivity disorder (ADHD), pervasive developmental disorder-not otherwise specified (PDD-NOS), Rett syndrome, Asperger syndrome, Angelman syndrome, generalized anxiety disorder, social anxiety disorder, bipolar disorder, epilepsy, oppositional defiant disorder, cerebral folate deficiency, FOLRl-related cerebral folate transport deficiency, or a sleep disorder. In some specific embodiments, the neurodevelopmental disorder may be autism spectrum disorder.

[0148] In some embodiments, the compositions described herein may improve or reduce symptoms associated with neurodevelopment disorders, e.g., autism spectrum disorder. For example, in some embodiments, symptoms associated with neurodevelopment disorder may include irritability, homing deficits, deficits in adaptive functioning, chronic pain, seizures, difficulty with communication, difficulty with social interactions, obsessive interests, repetitive behaviors, inappropriate social interaction, poor eye contact, compulsive behavior, impulsivity, repetitive movements, self-harm, persistent repetition of words or actions learning disabilities, speech delays, depression, anxiety, changes, tics, and sound sensitivity.

[0149] In some embodiments, the method of treating autism spectrum disorder in a subject may result in a reduction of the symptoms of ASD that is characterized by improvement according to the Vineland Adaptive Behavior Scales, ADOS-2, CGI-C social deficit subscales, aberrant behavior checklist, the Autism Treatment Evaluation Checklist (ATEC), Social Responsiveness Scale, 2ndEdition (SRS-2), or any combination thereof.

[0150] In some embodiments, the method of treating autism spectrum disorder in a subject may result in an improvement in the symptoms of ASD compared to prior to the treatment that is characterized by an improvement according to the Autism Behavior Inventory (ABI)-Social Communication Domain Score.

[0151] The ABI-Social Communication Domain Score is a 62-item questionnaire for reporting the behaviors of subjects (ages: 3 years-adulthood) diagnosed with ASD. The tool is suitable for completion by parents or care / study partners of people with ASD. Each item assesses either quality (from not at all to without help) or frequency (never to very often) of aparticular behavior. The Social Communication domain score is the sum of the scores in the social communication domain divided by the number of items in the domain.

[0152] In some embodiments, the method of treating autism spectrum disorder in a subject may result in an improvement in symptoms associated with ASD compared to prior to the treatment that is characterized by about a 20% improvement, about a 25% improvement, about a 30% improvement, about a 35% improvement, about a 40% improvement, about a 45% improvement, about a 50% improvement, about a 55% improvement, about a 60% improvement, about a 65% improvement, about a 70% improvement, about a 80% improvement, about a 90% improvement, or about a 95% improvement in ABI-Social Communication Domain Score compared to prior to treatment.

[0153] In some embodiments, the method of treating autism spectrum disorder in a subject may result in an improvement in symptoms associated with ASD compared to prior to the treatment that is characterized by at least about a 20% improvement, at least about a 25% improvement, at least about a 30% improvement, at least about a 35% improvement, at least about a 40% improvement, at least about a 45% improvement, at least about a 50% improvement, at least about a 55% improvement, at least about a 60% improvement, at least about a 65% improvement, at least about a 70% improvement, at least about a 80% improvement, at least about a 90% improvement, or at least about a 95% improvement in ABI-Social Communication Domain Score compared to prior to treatment.

[0154] In embodiments, the method of treating autism spectrum disorder in a subject may result in an improvement in the symptoms of ASD compared to prior to the treatment that is characterized by an improvement according to the Clinician Global Impression of Improvement (CGI-I) score.

[0155] The CGI-I score is a single-item instrument based on a 7-point scale used to capture the Investigator's global impression of response. The Investigator or designee rates the improvement ob served from 1-7 (1 = very much improved; 2 = much improved; 3 = minimally improved; 4 = no change; 5 = minimally worse; 6 = much worse; 7 = very much worse).

[0156] In embodiments, the method of treating autism spectrum disorder in a subject may result in an improvement in symptoms associated with ASD compared to prior to the treatment and has a CGI-I score of about 1, about 2, about 3, about 4, or about 5 after-51-treatment. In embodiments, after said treatment the patient experiences an improvement in symptoms associated with ASD and has a CGI-I score of about 1, about 2, or about 3. In embodiments, the method of treating autism spectrum disorder in a subject may result in an improvement in symptoms associated with ASD and has a CGI-I score of about 1 or 2.

[0157] In some embodiments, the method of treating autism spectrum disorder in a subject may result in an improvement in symptoms associated with ASD that is characterized by improvement according to the Autism Behavior Inventory-Clinician (ABI-C) Score compared to prior to the treatment.

[0158] The ABI-Clinician (ABI-C) captures the clinician rating of behaviors of a person with ASD that occurred over the week prior to assessment. It contains 14 items reflecting the core and associated autism behavior domains: Social Communication, Restrictive Behaviors, Mood and Anxiety, Self Regulation, and Challenging Behavior. Each item is rated on a 7-point scale from 1 (none; no symptoms present) to 7 (very severe; persistent interference with function or adaptation).

[0159] In some embodiments, the method of treating autism spectrum disorder in a subject may result in an improvement in symptoms compared to prior to the treatment that is characterized by about a 20% improvement, about a 25% improvement, about a 30% improvement, about a 35% improvement, about a 40% improvement, about a 45% improvement, about a 50% improvement, about a 55% improvement, about a 60% improvement, about a 65% improvement, about a 70% improvement, about a 80% improvement, about a 90% improvement, or about a 95% improvement in ABI-C score compared to prior to treatment.

[0160] In some embodiments, the method of treating autism spectrum disorder in a subject may result in an improvement in symptoms compared to prior to the treatment that is characterized by at least about a 20% improvement, at least about a 25% improvement, at least about a 30% improvement, at least about a 35% improvement, at least about a 40% improvement, at least about a 45% improvement, at least about a 50% improvement, at least about a 55% improvement, at least about a 60% improvement, at least about a 65% improvement, at least about a 70% improvement, at least about a 80% improvement, at least about a 90% improvement, or at least about a 95% improvement in ABI-C score compared to prior to treatment.

[0161] In some embodiments, the method of treating autism spectrum disorder in a subject may result in an improvement in the symptoms of ASD that is characterized by improvement according to the ABI Repetitive / Restrictive Behavior Domain Score compared to prior to the treatment.

[0162] Each item on the ABI-Repetitive / Restrictive Behavior Domain is rated by frequency (0=never to 3=very often). The domain score is the sum of all domain items scores divided by the number of items in the domain.

[0163] In some embodiments, the method of treating autism spectrum disorder in a subject may result in an improvement in the symptoms of ASD that is characterized by improvement according to the ABI Challenging Behavior Domain Score compared to prior to the treatment.

[0164] Each item on the ABI-Challenging Behavior Domain is rated by frequency (0=never to 3=very often). The domain score is the sum of all domain items scores divided by the number of items in the domain.

[0165] In some embodiments, the method of treating autism spectrum disorder in a subject may result in an improvement in the symptoms of ASD that is characterized by improvement according to the ABI-Short Form (ABI-S) Score compared to prior to the treatment.

[0166] The ABI-S is a 24-item short version of the ABI, containing items from each of the five domains. The domain score for each domain is calculated as the sum of scores all domain items divided by the number of items in the domain.

[0167] In some embodiments, the method of treating autism spectrum disorder in a subject may result in experiences an improvement in the symptoms of ASD that is characterized by improvement according to the ABC-Social Withdrawal (ABC-SW) Subscale Score compared to prior to the treatment.

[0168] The ABC-SW subscale consists of 16 items of the ABC-2 rated from 0 (not a problem) to 3 (the problem is severe in degree).

[0169] In some embodiments, the method of treating autism spectrum disorder in a subject may result in an improvement in the symptoms of ASD that is characterized by improvement according to Vineland-3 (Domain Level Version) Score: total ofCommunication, Socialization, and Maladaptive behavior domains compared to prior to the treatment,

[0170] The Vineland-3 Domain Level Version contains 5 domains. Responses on each item are rated from 0 (never) to 2 (usually).

[0171] The Vineland Adaptive Behavior Scales (VABS), third edition, is a standardized measure of adaptive behavior used to evaluate the personal and social skills of an individual from birth through adulthood. Individuals can be evaluated on the VABS scale by either teachers or caregivers. According to the VABS, an individual is assigned a VABS adaptive behavior composite score, which measures an individual’s functioning compared to others of his or her age. An individual is also assigned domain scores in communication, daily living skills, socialization, and motor skills to assess an individual’s adaptive behavior strengths and weaknesses. An individual receives a score from 20 to 140 on each of the domain scores and the VABS adaptive behavior composite score. A score from 20 to 70 represents low- adaptive level. A score from 71 to 85 represents moderately low adaptive level. A score from 86 to 114 represents moderately adequate adaptive level. A score from 115 to 129 represents moderately high adaptive level. A score from 130 to 140 represents high adaptive level.

[0172] In some embodiments, the method of treating autism spectrum disorder in a subject may result in a reduction of symptoms associated with ASD that is characterized by at least a one point increase in the VABS adaptive behavior composite score compared to prior to the treatment. In some embodiments, the increase in the VABS adaptive behavior score is about 1 point, about 2 points, about 3 points, about 4 points, about 5 points, about 6 points, about 7 points, about 8 points, about 9 points, about 10 points, about 11 points, about 12 points, about 13 points, about 14 points, about 15 points, about 16 points, about 17 points, about 18 points, about 19 points, about 20 points, about 21 points, about 22 points, about 23 points, about 24 points, about 25 points, about 26 points, about 27 points, about 28 points, about 29 points, about 30 points, about 31 points, about 32 points, about 33 points, about 34 points, about 35 points, about 36 points, about 37 points, about 38 points, about 39 points, about 40 points, about 41 points, about 42 points, about 43 points, about 44 points, about 45 points, about 46 points, about 47 points, about 48 points, about 49 points, about 50 points, about 51 points, about 52 points, about 53 points, about 54 points, about 55 points, about 56 points, about 57 points, about 58 points, about 59 points, about 60 points, about 61 points, about 62points, about 63 points, about 64 points, about 65 points, about 66 points, about 67 points, about 68 points, about 69 points, or about 70 points compared to prior to said treating.

[0173] In some embodiments, the increase in VABS adaptive behavior composite score is at least about 1 %, at least about 5 %, at least about 10 %, at least about 15 %, at least about 20 %, at least about 25 %, at least about 30 %, at least about 35 %, at least about 40 %, at least about 45 %, at least about 50 %, at least about 55 %, or at least about 60 % compared to prior to said treating.

[0174] In some embodiments, the method of treating autism spectrum disorder in a subject may result in an increase in sociability that is characterized by an increase in the VABS adaptive behavior socialization domain score compared to prior to the treatment. In some embodiments, the increase in the VABS adaptive behavior socialization domain score is about 1 point, about 2 points, about 3 points, about 4 points, about 5 points, about 6 points, about 7 points, about 8 points, about 9 points, about 10 points, about 11 points, about 12 points, about 13 points, about 14 points, about 15 points, about 16 points, about 17 points, about 18 points, about 19 points, about 20 points, about 21 points, about 22 points, about 23 points, about 24 points, about 25 points, about 26 points, about 27 points, about 28 points, about 29 points, about 30 points, about 31 points, about 32 points, about 33 points, about 34 points, about 35 points, about 36 points, about 37 points, about 38 points, about 39 points, about 40 points, about 41 points, about 42 points, about 43 points, about 44 points, about 45 points, about 46 points, about 47 points, about 48 points, about 49 points, about 50 points, about 51 points, about 52 points, about 53 points, about 54 points, about 55 points, about 56 points, about 57 points, about 58 points, about 59 points, about 60 points, about 61 points, about 62 points, about 63 points, about 64 points, about 65 points, about 66 points, about 67 points, about 68 points, about 69 points, or about 70 points compared to prior to said treating.

[0175] In some embodiments, the increase in VABS adaptive behavior socialization domain score is at least about 1 %, at least about 5 %, at least about 10 %, at least about 15 %, at least about 20 %, at least about 25 %, at least about 30 %, at least about 35 %, at least about 40 %, at least about 45 %, at least about 50 %, at least about 55 %, or at least about 60 %, compared to prior to said treating. In some embodiments, the patient experiences an at least 10 % improvement on the socialization domain score of VABS after treatment. Insome embodiments, the subject experiences an at least 35 % improvement on the socialization domain score of VABS after treatment

[0176] In some embodiments, the method of treating autism spectrum disorder in a subject may result in an improvement in communication that is characterized by an increase in the VABS adaptive behavior communication domain score compared to prior to the treatment. In some embodiments, the increase in the VABS adaptive behavior communication domain is about 1 point, about 2 points, about 3 points, about 4 points, about 5 points, about 6 points, about 7 points, about 8 points, about 9 points, about 10 points, about 11 points, about 12 points, about 13 points, about 14 points, about 15 points, about 16 points, about 17 points, about 18 points, about 19 points, about 20 points, about 21 points, about 22 points, about 23 points, about 24 points, about 25 points, about 26 points, about 27 points, about 28 points, about 29 points, about 30 points, about 31 points, about 32 points, about 33 points, about 34 points, about 35 points, about 36 points, about 37 points, about 38 points, about 39 points, about 40 points, about 41 points, about 42 points, about 43 points, about 44 points, about 45 points, about 46 points, about 47 points, about 48 points, about 49 points, about 50 points, about 51 points, about 52 points, about 53 points, about 54 points, about 55 points, about 56 points, about 57 points, about 58 points, about 59 points, about 60 points, about 61 points, about 62 points, about 63 points, about 64 points, about 65 points, about 66 points, about 67 points, about 68 points, about 69 points, or about 70 points, compared to prior to the treatment.

[0177] In some embodiments, the increase in VABS adaptive behavior communication domain score is at least about 1 %, at least about 5 %, at least about 10 %, at least about 15 %, at least about 20 %, at least about 25 %, at least about 30 %, at least about 35 %, at least about 40 %, at least about 45 %, at least about 50 %, at least about 55 %, or at least about 60 % compared to prior to said treating.

[0178] The Autism Diagnostic Observation Schedule, Second Edition (ADOS-2) is an instrument that allows accurate assessment and diagnosis of ASD across age, developmental level, and language skills. The ADOS-2 is a clinician-administered observational assessment that comprises two behavioral domains: social affect (SA) and restricted and repetitive behaviors (RRB). An individual is administered one of the five ADOS-2 modules, which is selected on the basis of the individual’s expressive language level and chronological age. The toddler module is for children between 12 and 30 months of age whodo not consistently use phrase speech. Module 1 is for children 31 months and older who do not consistently use phrase speech. Module 2 is for children of any age who use phrase speech but are not verbally fluent. Module 3 is for verbally fluent children and young adolescents. Module 4 is for verbally fluent older adolescents and adults. An individual is assigned an ADOS-2 Composite Total score ranging from 1 to 10. Individuals with ASD are characterized by an ADOS-2 Composite Total score between 6 and 10.

[0179] In some embodiments, the method of treating autism spectrum disorder in a subject may result in a reduction of symptoms associated with ASD associated with an at least one point decrease in the ADOS-2 Composite Total score compared to prior to said treating. In some embodiments, the patient experiences an improvement in the symptoms of ASD that is characterized by a decrease of at least 1 point, at least 2 points, at least 3 points, at least 4 points, or at least 5 points on the ADOS-2 Composite Total score compared to prior to said treating.

[0180] In some embodiments, the subject may experience an improvement in the symptoms of ASD that is characterized by an at least about 5 %, or at least about 10 %, or at least about 15 %, or at least about 20 %, or at least about 25 %, or at least about 30 %, or at least about 35 %, or at least about 40 %, or at least about 45 %,or at least about 50 %, or at least about 55 %, or at least about 60 %, or at least about 65 %, or at least about 70 %, or at least about 75 %, or at least about 80 %, or at least about 85 %, or at least about 90 % improvement in the ADOS-2 composite score compared to prior to said treating.

[0181] In some embodiments, the subject may experience a reduction of symptoms associated with ASD that is characterized by an at least one point decrease in the ADOS-2 module 4 social affect score compared to prior to said treatment. In some embodiments, the decrease in the ADOS-2 module 4 social affect score is correlated with an improvement in sociability. In some embodiments, the improvement in sociability is characterized by an at least 1 point, or at least 2 point, or at least 3 point, or at least 4 point, or at least 5 point, or at least 6 point decrease in the social affect score on the ADOS-2 module 4 compared to prior to said treating.

[0182] In some embodiments, the improvement in sociability is characterized by an at least 10 %, or at least 15 %, or at least 20 %, or at least 25 %, or at least 30 %, or at least 35 %, or at least 40 %, or at least 45 %, or at least 50 %, or at least 55 %, decrease in the socialaffect scale on ADOS-2 module 4 compared to prior to said treating. In some embodiments, the patient experiences an improvement in sociability that is characterized by an at least 10 % decrease in the social reciprocity score on the Autism Diagnostic Observation Schedule module 4 compared to prior to said treating.

[0183] The clinical global impression-severity (CGI-S) scale is utilized by clinicians to rate the severity of an ASD patient’s symptoms. An individual is assigned a score of 1 to 7. A score of 1 represents a normal patient. A score of 7 represents the score of a patient with ASD.

[0184] In some embodiments, the method of treating autism spectrum disorder in a subject may result in a reduction of symptoms associated with ASD that is characterized by at least a one point decrease in the CGI-S score compared to prior to said treating. In some embodiments, the patient experiences an at least 1 point, or at least 2 points, or at least 3 points, or at least 4 points, or at least 5 points decrease in the CGI-S scale compared to prior to said treating.

[0185] In some embodiments, the subject may experience an at least 5 %, or at least 10 %, or at least 15 %, or at least 20 %, or at least 25 %, or at least 30 %, or at least 35 %, or at least 40 %, or at least 45 %, or at least 50 %, or at least 55 %, or at least 60 %, or at least 65 %, or at least 70 %, or at least 75 %, or at least 80 %, or at least 85 %, or at least 90 % reduction in the CGI-S scale compared to prior to said treating.

[0186] The clinical global impression-severity (CGI-C) scale is utilized by clinicians to evaluate a change in an ASD patient’s symptoms. An individual is assigned a score from 1 (very' much improved) to 7 (very much worse).

[0187] In some embodiments, after said treatment the subject may experience a reduction of symptoms associated with ASD that is characterized by at least a one point decrease in the CGI-C scale. In some embodiments, the patient experiences an improvement in irritability that is characterized by a CGI-C score of < 1, < 2, < 3, or < 4 after said treatment. In some embodiments, the patient’s improvement in irritability that is characterized by a Clinical Global Impression-Change (CGI-C) score of < 3 after said treatment. In some embodiments, the patient experiences an improvement in sociability that is characterized by a CGI-C score of < 1, < 2, < 3, or < 4 after said treatment. In some embodiments, the patientexperiences an improvement in sociability that is characterized by a CGI-C score of < 3 after said treatment.

[0188] The Autism Behavior Inventory (ABI) social deficit subscale is a measure used for assessing changes in the core and associated symptoms of ASD. An individual is administered the ABI-full (93 items) or ABI-short version (36) scales. An individual is scored from 0 points to 6 points on each item of the scales, where 0 is the absence of symptoms and 6 is maximum symptoms. An individual that receives a score of 0 does not exhibit symptoms. An individual that is assigned a score of 6 experiences severe ASD symptoms.

[0189] In some embodiments, the subject may experience an improvement in symptoms that is characterized by a decrease in ABI score of at least one point. In some embodiments, the patient experiences an improvement in symptoms that is characterized by a decrease in ABI score of at least 1 point, or at least 2 points, or at least 3 points, or at least 4 points, compared to prior to said treating.

[0190] In some embodiments, the subject may experience an improvement in symptoms that is characterized by an at least 5 %, or at least 10 %, or at least 15 %, or at least 20 %, or at least 25 %, or at least 30 %, or at least 35 %, or at least 40 %, or at least 45 %, or at least 50 %, or at least 55 %, or at least 60 %, or at least 65 %, or at least 70 %, or at least 75 %, or at least 80 %, or at least 85 %, or at least 90 %, or at least 95 % reduction in ABI score compared to prior to said treating.

[0191] The childhood autism rating scale (CARS) is used to identify children two years and older with ASD. The CARS consists of 14 domains assessing behaviors associated with ASD, with a 15th domain rating general impressions of autism. Each domain is scored on a scale ranging from one to four; higher scores are associated with a higher level of impairment. Total scores can range from a low of 15 to a high of 60; scores below 30 indicate that the individual is in the non-autistic range, scores between 30 and 36.5 indicate mild to moderate autism, and scores from 37 to 60 indicate severe autism.

[0192] In some embodiments, the method of treating autism spectrum disorder in a subject may result in a reduction of symptoms associated with ASD that is characterized by a decrease in the CARS total score of about 1 point, about 2 points, about 3 points, about 4 points, about 5 points, about 6 points, about 7 points, about 8 points, about 9 points, about 10 points, about 11 points, about 12 points, about 13 points, about 14 points, about 15 points,about 16 points, about 17 points, about 18 points, about 19 points, about 20 points, about 21 points, about 22 points, about 23 points, about 24 points, about 25 points, about 26 points, about 27 points, about 28 points, about 29 points, or about 30 points, compared to prior to said treating.

[0193] In some embodiments, the method of treating autism spectrum disorder in a subject may result in a reduction of symptoms associated with ASD that is characterized by a decrease in the CARS total score of at least 5 %, or at least 10 %, or at least 15 %, or at least 20 %, or at least 25 %, or at least 30 %, or at least 35 %, or at least 40 %, or at least 45 %, or at least 50 %, or at least 55 %, or at least 60 %, or at least 65 %, or at least 70 %, or at least 75 %, or at least 80 %, or at least 85 %, or at least 90 %, or at least 95 %, compared to prior to said treating.

[0194] The social responsiveness scale, 2”dEdition (SRS-2) is utilized to obtain an efficient quantitative measure of the various dimensions of interpersonal behavior, communication, and repetitive / stereotypic behavior associated with ASD. According to the SRS-2, an individual is assigned a proxy version total t-score, based on the SRS-2. An individual is assigned a proxy version t-score which reflects the sum of responses to 65 social responsiveness scale questions, which serves as an index of severity of social skills across the autism spectrum. An individual that receives a proxy version t-score of greater than 76 receives a severe clinical diagnosis of ASD. If an individual is assigned a proxy version t-score of between 66 and 75, which is associated with moderate deficiencies in reciprocal social behavior that are clinically significant and lead to a substantial interference in everyday social interactions. If the individual is assigned a proxy version t-score between 60 to 65, the individual exhibits mild to moderate deficits in social interaction. If a patient exhibits a proxy version t-score of 59 or below, the patient exhibits normal social interactions.

[0195] In some embodiments, prior to administering of a composition described herein to a subject in need thereof, the subject in need thereof exhibits a proxy version total t-score of > 66.

[0196] In some embodiments, the subject may an improvement in symptoms that is characteri zed by a decrease in proxy version total t-score of at least one point, or at least two points, or at least three points, or at least four points, or at least five points, or at least six points, or at least seven points, or at least eight points, or at least nine points, or at least ten points, orat least eleven points, or at least twelve points, or at least thirteen points, or at least fourteen points, or at least fifteen points, or at least sixteen points, or at least seventeen points, or at least eighteen points, or at least twenty points, or at least twenty one points, or at least twenty two points, or at least twenty three points, or at least twenty four points, or at least twenty five points, compared to prior to said treating. In some embodiments, the subject may experience an improvement in symptoms that is characterized by a decrease in proxy version total t-score of the SRS-2 of at least 5 %, or at least 10 %, or at least 15 %, or at least 20 %, or at least 25 %, or at least 30 %, or at least 35 %, or at least 40 %, or at least 45 %, or at least 50 %, or at least 55 %, or at least 60 %, or at least 65 %, or at least 70 %, or at least 75 %, or at least 80 %, or at least 85 %, or at least 90 %, or at least 95 %, compared to prior to said treating.

[0197] The social responsiveness scale for adults (SRS-A) is a tool to evaluate social responsiveness in adulthood. The SRS-A contains 65 items which are scored from 0 to 3. If an individual receives a score of 0, the individual does not have the symptom. If an individual receives a score of 3, the individual has a severe symptom. An SRS-A total score of 67 indicates that an individual has ASD. The maximum SRS-A total score is 195.

[0198] In some embodiments, the method of treating autism spectrum disorder in a subject may result in a reduction in symptoms associated with ASD that is characterized by a decrease in the SRS-A score of about 5 %, about 10 %, about 15 %, about 20 %, about 25 %, about 30 %, about 35 %, about 40 %, about 45 %, about 50 %, about 55 %, about 60 %, about 65 %, about 70 %, compared to prior to said treatment, embodiments, the method of treating autism spectrum disorder in a subject may result in a reduction in symptoms associated with ASD that is characterized by an at least 10 % decrease in the SRS- A score compared to prior to said treatment.

[0199] In some embodiments, an improvement in sociability is associated with a decrease in the SRS-A score of about 10 %, about 15 %, about 20 %, about 25 %, about 30 %, about 35 %, about 40 %, or about 50 %, compared to prior to said treatment.

[0200] The aberrant behavior checklist (ABC) is a behavior rating scale that is utilized to rate individuals in five subscales: (1) irritability, agitation, and crying, (2) lethargy, social withdrawal, (3) stereotypic behavior, (4) hyperactivity and noncompliance, and (5) inappropriate speech. Individuals can be evaluated on the ABC by any adult that knows the individual well. The ABC contains a 58-item questionnaire that is utilized to assess the five-subscales. Each item on the 58-item questionnaire is rated from 0 to 3. A score of 0 means an absence in symptom. A score of 3 means an individual experiences the symptom with high severity. Items within each subscale are added to obtain a subscale score. Possible subscale scores on the ABC range from 0 to 48 with higher sub-scores indicating behavioral impairment.

[0201] In some embodiments, after said treatment the subject may experience a reduction of symptoms associated with ASD that is characterized by at least a one point decrease in the ABC irritability agitation and crying (ABC-I) sub-score. In some embodiments, after said treatment the patient experiences a reduction in irritability that is characterized by a decrease in the ABC-I communication domain score compared to prior to the treatment.

[0202] In some embodiments, the subject exhibits an ABC-I score of > 18 prior to treatment.

[0203] In some embodiments, the method of treating autism spectrum disorder in a subject may result in a reduction in irritability that is characterized by an about 2 point, about 4 point, about 6 point, about 8 point, about 10 point, about 15 point, about 20 point, about 25 point, or about 30 point decrease in the ABC-I domain score compared to prior to said treatment. In some embodiments, the decrease in the ABC-I domain score is about 5 %, about 10 %, about 15 %, about 20 %, about 25 %, about 30 %, about 35 %, about 40 %, about 45 %, about 50 %, about 55 %, about 60 %, about 65 %, about 70 %, about 75 %, about 80 %, about 85 %, about 90 %, or about 95 % compared to prior to said treatment. In some embodiments, the decrease in the ABC-I domain score is at least about 5 %, at least about 10 %, at least about 15 %, at least about 20 %, at least about 25 %, at least about 30 %, at least about 35 %, at least about 40 %, at least about 45 %, at least about 50 %, at least about 55 %, at least about 60 %, at least about 65 %, at least about 70 %, at least about 75 %, at least about 80 %, at least about 85 %, at least about 90 %, or about at least 95 % compared to prior to said treatment.

[0204] In embodiments, the method of treating autism spectrum disorder in a subject may result in a reduction in irritability that is characterized by a 25% reduction on the irritability subscale and wherein the patient is rated as much improved or very much improved (i.e., 2 or 1, respectively) on the CGI improvement scale.

[0205] In some embodiments, after said treatment the subject may experience a reduction of symptoms associated with ASD that is characterized by at least a one-point decrease in the ABC-Lethargy and Social Withdrawal (ABC-LSW) subscore compared to priorto said treatment. In some embodiments, after said treatment the subject may experience a reduction in lethargy and social withdrawal that is characterized by a decrease in the ABC-LSW communication domain score compared to prior to the treatment.

[0206] In some embodiments, the decrease in the ABC-LSW communication domain score is about 2 points, about 4 points, about 6 points, about 8 points, about 10 points, about 15 points, about 20 points, about 25 points, or about 30 points, compared to prior to said treatment.

[0207] In some embodiments, the decrease in the ABC-LSW communication domain score is about 5 %, about 10 %, about 15 %, about 20 %, about 25 %, about 30 %, about 35 %, about 40 %, about 45 %, about 50 %, about 55 %, about 60 %, about 65 %, about 70 %, about 75 %, about 80 %, about 85 %, about 90 %, or about 95 %, compared to prior to said treatment.

[0208] The social communication questionnaire is a tool used by physicians to screen patients with ASD. An individual’s caregiver rates the verbal individual on a score of 0 to 39 or the non-verbal individual on a scale of 0 to 33. An individual that has ASD is characterized by a social communication questionnaire that is above 15.

[0209] In some embodiments, the method of treating autism spectrum disorder in a subject may result in a reduction of symptoms associated with ASD that is characterized by an at least 10 %, at least 15 %, at least 20 %, at least 25 %, at least 30 %, at least 35 %, at least 40 %, at least 45 %, at least 50 %, at least 55 %, at least 60 %, at least 65 %, at least 70 %, at least 75 %, at least 80 %, at least 85 %, at least 90 %, or at least 95 % decrease in the social communication questionnaire scale compared to prior to said treatment.

[0210] The pervasive developmental disorder behavior inventory (PDDBI) is utilized to evaluate the efficacy of the treatment of ASD. The PDDBI assesses both problem behaviors and appropriate social, language, and learning / memory skills. The PDDBI is divided into two behavioral dimensions: (a) approach- withdrawal problems, assessing maladaptive behaviors and (b) receptive / expressive social communication abilities, assessing social communication competence. The approach-withdrawal problem dimension is further divided into behavioral domains, including sensory / perceptual approach behaviors, ritualisms / resistance to chain (RITUAL), social pragmatic problems (SOCPP), semantic / pragmatic problems (SEMPP), arousal regulation problems (AROUSE), specificfears (FEARS), and aggressiveness (AGG) domains. The receptive / expressive social communication abilities, assessing social communication competence (REXSCA) is further divided into behavioral domains, including the social approach behaviors (SOCAPP), expressive language (EXPRESS), and learning memory and receptive language (LMRL) domains. An individual receives a T score for each domain and a composite score, which is the sum of the scores in each domain. An individual with ASD is assigned a T score of greater than 50.

[0211] In some embodiments, the pervasive developmental disorder behavior inventory (PDDBI) is utilized to characterize the reduction in symptoms associated with ASD provided by the methods of the disclosure.

[0212] In some embodiments, a subject may exhibit a T score of greater than 50 prior to said treatment. In some embodiments, after said treatment the subject may experience a reduction of symptoms associated with ASD that is characterized by a decrease in the PDDBI by about 5 %, or about 10 %, or about 15 %, or about 20 %, or about 25 %, or about 30 %, or about 35 %, or about 40 %, or about 45 %, or about 50 %, or about 55 %, or about 60 %, or about 65 %, or about 70 %, or about 75 %, or about 80 %, or about 85 %, or about 90 %, or about 95 % compared to prior to said treating.

[0213] The ATEC is a scale utilized to evaluate the efficacy of ASD treatments. The ATEC is a one-page form designed to be completed by parents, teachers, or caretakers. The ATEC consists of four subtests: speech / language communication, sociability, sensory / cognitive awareness, and health / physical / behavior. An individual is assigned a speech / language communication subtest rating from 0 to 28. An individual is assigned a sociability subtest rating from 0 to 40. An individual is assigned a sensory / cognitive awareness subtest rating from 0 to 36. An individual is assigned a health / physical / behavior subtest rating of 0 to 75. The individual subsets are summed, and the individual can receive a maximum score of 180 points

[0214] In some embodiments, after said treatment the subject may experience a reduction of symptoms associated with ASD that is characterized by a decrease in the mean ATEC score compared to prior to the treatment. In some embodiments, after said treatment the subject may experience an improvement in speech / language communication that is characterized by a decrease in the speech / language communication subtest rating compared toprior to said treating. In some embodiments, after said treatment the subject may experience an improvement in sociability that is characterized by a decrease in the sociability subtest rating compared to prior to said treating. In some embodiments, after said treatment the subject may experience an improvement in sensory / cognitive awareness that is characterized by a decrease in sensory / cognitive awareness subtest rating compared to prior to said treating. In some embodiments, after said treatment the subject may experience an improvement in health / physical / behavior that is characterized by a decrease in health / physical / behavior rating, compared to prior to said treating.

[0215] In some embodiments, the subject may exhibit a decrease on the sociability subsection of the ATEC of at least 1 point, or about 2 points, or about 3 points, or about 4 points, or about 5 points, or about 6 points, or about 7 points, or about 8 points, or about 9 points, or about 10 points, or about 11 points, or about 12 points, or about 13 points, or about 14 points, or about 15 points, or about 16 points, or about 17 points, or about 18 points, or about 19 points, or about 20 points compared to prior to said treating. In some embodiments, the subject may exhibit a decrease on the sociability subsection of the ATEC of at least 1 point compared to prior to said treating.

[0216] In some embodiments, the subject may exhibit a decrease on the sociability subsection of the ATEC of at least 5 %, or at least 10 %, or at least 15 %, or at least 20 %, or at least 25 %, or at least 30 %, or at least 35 %, or at least 35 %, or at least 40 %, or at least 50 % compared to prior to said treating. In some embodiments, the subject may exhibit a decrease on the sociability subsection of the ATEC of at least 10 % compared to prior to said treating.

[0217] Objective / performance-based assessments are utilized to measure the reduction in the symptoms associated with ASD after administering a therapeutically effective amount of a triptan to a patient in need thereof. In some embodiments, the objective / performance based assessments are selected from the group consisting of eye gaze tracking of social stimuli (eye tracking), parent engagement rating inventory (JERI), and Noldus Ethovision Analysis, In some embodiments, eye gaze tracking of social stimuli is utilized to characterize the reduction in symptoms associated with ASD provided by the methods of the disclosure. Patients with ASD exhibit significantly shorter look durations to the eyes (eye contact) as compared to their peers without ASD. Patients with ASD have moredifficulty locating and processing pertinent social information the more rapidly stimuli are presented.

[0218] The PCIT provides training methods designed to help adults improve their parenting and language skills and to help children learn how to better control emotions. In some embodiments, the parent child interaction task (PCIT) is utilized to improve the relationship between the caregiver and the patient with ASD. In some embodiments, the PCIT is helpful for reducing child behavior issues and increasing communication and interaction skills within the family. In some embodiments, children who participate in CPT develop greater self-esteem, experience less anger and frustration, see an improvement in social, organizational, and play skills, feel safer and calmer, and communicate more effectively.

[0219] The Joint Engagement Ranking Inventory (JERI) is a tool which is utilized to measure targets for early intervention for developmental disorders and delays, such as ASD. The JERI is an 18-item inventory developed to measure relevant intervention targets, such as unengagement, object engagement, joint engagement, stereotyped, restricted and repetitive behavior, attention to the caregiver, initiation of communication, expressive language level and use, scaffolding, following-in, caregiver affect, fluency and connectedness, and shared routines and rituals. An individual evaluated on the JERI scale receives a score from 1 to 7 on each item in the JERI inventory. A score of 7 is assigned to an individual with the most joint engagements. A score of 1 on an item is assigned to an individual that has no episodes of joint engagement. A lower JERI scale is correlated with ASD.

[0220] In some embodiments, a reduction in the symptoms in ASD is associated with lower scores on the JERI compared to prior to treatment by the methods of the disclosure. In some embodiments, after said treatment the patient experiences a reduction of symptoms associated with ASD that is characterized by a decrease in the JERI score compared to prior to the treatment. In some embodiments, the JERI scores are reduced by about 10 %, about 20 %, about 30 %, about 40 %, about 50 %, about 60 %, or about 70 % compared to prior to the treatment.

[0221] The Ohio State University Autism Rating Scale (OARS-5) measures persistent impairment in social interactions, restrictive interests / activities and repetitions in behavior, and level of support from social, academic, and community (Hollway, J. A., Arnold, L. E., & Aman, M. G. (2017, September). OSU Autism Rating Scale -DSM-5 (OARS-5).). TheOARS-5 was developed to provide three types of summary scores: (A) a Autism Symptom Count, based on clinical interview (OARS-5 Total Score); (b) a Weighted Mean Severity score based on severity of autism symptoms, derived from the clinical interview; (c) and an Impairment Index ranging from 0 to 9, based on the level of supports needed due to severity. The OARS-5 Total Score is equal to OARS-5 Social Deficits Subscale Score + OARS-5 Restricted Patterns of Interest Subscale Score.

[0222] The OARS-5 includes the following subscales:

[0223] Section A: Persistent impairment in social interactions across multiple settings (OARS-5 Social Deficits Subscale Score);

[0224] Section B: Restricted interests / activities and repetitive patterns of behavior (OARS- 5 Restricted Patterns of Interest Subscale Score) and

[0225] Section C: Level of support for Section A (social interactions / communication) and B (restricted and repetitive behaviors).

[0226] In some embodiments, after said treatment the subject may experience a reduction of symptoms associated with ASD that is characterized by a decrease in the OARS-5 Total Score compared to prior to the treatment. In some embodiments, after said treatment the subject may experience a reduction of symptoms associated with ASD that is characterized by a decrease of about 1 point, about 2 points, about 3 points, or about 4 points, about 5 points, about 6 points, about 7 points, about 8 points, about 9 points or about 10 points in the OARS-5 Total Score compared to prior to the treatment.

[0227] In some embodiments, after said treatment the subject may experience a reduction of symptoms associated with ASD that is characterized by a decrease in the OARS-5 Social Deficits Subscale Score compared to prior to the treatment. In some embodiments, after said treatment the subject may experience a reduction of symptoms associated with ASD that is characterized by a decrease of about 1 point, about 2 points, about 3 points, or about 4 points, about 5 points, about 6 points, about 7 points, about 8 points or about 9 points in the OARS-5 Social Deficits Subscale Score compared to prior to the treatment.

[0228] In some embodiments, after said treatment the patient experiences a reduction of symptoms associated with ASD that is characterized by a decrease in the Ohio State University Autism Clinical Global Impression (OSU Autism CGI) total score compared to prior to the treatment. OSU Autism CGI-S total score is equal to OSU Autism CGI SeverityScale (OSU Autism CGI-S) Score + OSU Autism CGI-Improvement Scale (OSU Autism CGI-I) Score.

[0229] In some embodiments, after said treatment the subject may experience a reduction of symptoms associated with ASD that is characterized by a decrease of about 1 point, about 2 points, about 3 points, or about 4 points, about 5 points, about 6 points, or about 7 points in the OSU Autism CGI total score compared to prior to the treatment.

[0335] In some embodiments, after said treatment the patient experiences a reduction of symptoms associated with ASD that is characterized by a decrease in the Ohio State University Autism Clinical Global Impression-Improvement Scale (OSU Autism CGI-I) score compared to prior to the treatment. In some embodiments, after said treatment the patient experiences a reduction of symptoms associated with ASD that is characterized by a decrease of about 1 point, about 2 points, about 3 points, or about 4 points, about 5 points, about 6 points, or about 7 points in the OSU Autism CGI-I compared to prior to the treatment.

[0230] In some embodiments, after said treatment the subject may experience a reduction of symptoms associated with ASD that is characterized by a decrease in the Ohio State University Autism Clinical Global Impression-Severity Scale (OSU Autism CGI-S) score compared to prior to the treatment. In some embodiments, after said treatment the patient experiences a reduction of symptoms associated with ASD that is characterized by a decrease of about 1 point, about 2 points, about 3 points, or about 4 points, about 5 points, about 6 points, or about 7 points in the OSU Autism CGI-S compared to prior to the treatment.

[0231] In some embodiments, the present disclosure provides methods of treating the symptoms of ASD comprising administering an effective amount of a composition of the present disclosure to a subject in need thereof.

[0232] In embodiments, the subject is an adolescent or adult with an autism spectrum disorder (ASD). In embodiments, the subject is at least 12 years old. In embodiments, the subject is 12-45 years old.

[0233] In some embodiments, the subject in need thereof is a subject diagnosed with ASD according to the DSM-5 diagnostic criteria.

[0234] In some embodiments, prior to treatment according to the methods of the disclosure, the ASD patient in need thereof has a full scale IQ score on the Weschsler Abbreviated Scale of Intelligence (WASI®)-II of > 70.

[0235] In some embodiments, the present disclosure provides methods of treating the symptoms associated with ASD that are refractory' to treatment with existing ASD treatments. In some embodiments, the symptoms associated with ASD are refractory to treatment with aripiprazole. In some embodiments, the symptoms associated with ASD are refractory to treatment with risperidone.

[0236] In some embodiments, the treated ASD patient exhibits atypical sensory processing. Sensory processing refers to the ability to register, process, and organize sensory information and to execute appropriate responses to environmental demands, which manifest as hypersensitivities or hyposensitivities to stimuli. The patient in need with atypical sensory processing may have aversions to the color, taste, smell, and / or texture of foods or medicines. In some embodiments, atypical sensory processing is manifested in the patient in need as non-compliance with taking medications. In some embodiments, the treated ASD patient refuses to sw'allow' medications. In some embodiments, said medications are liquid formulations or pills.

[0237] In some embodiments, the treated ASD patient is an infant. In some embodiments, the treated ASD patient is a child. In some embodiments, the treated ASD patient is an adolescent. In some embodiments, the treated ASD sujbect is an adult. In some embodiments, the treated ASD subject is a geriatric patient.

[0238] In some embodiments, provided herein is a method of reducing a a fem ale's risk of having a fetus with a neurodevelopmental disorder, said method comprising administering to a subject in need thereof an effective amount of any one of the compositions described herein.

[0239] In some embodiments, the neurodevelopmental disorder may be fragile X syndrome, autism spectrum disorder, attention deficit hyperactivity disorder (ADHD), pervasive developmental disorder-not otherwise specified (PDD-NOS), Rett syndrome, Asperger syndrome, Angelman syndrome, generalized anxiety disorder, social anxiety disorder, bipolar disorder, epilepsy, oppositional defiant disorder, cerebral folate deficiency, FOLRl-related cerebral folate transport deficiency, or a sleep disorder. In some specific embodiments, the neurodevelopmental disorder may be autism spectrum disorder. In some embodiments, the method comprises administering the compositions described herein at preconception, during the first trimester of pregnancy, during the second trimester of pregnancy, and / or during the third trimester of pregnancy.

[0240] In some embodiments, the subject is a human.

[0241] In some embodiments, the composition is administered orally as a liquid or solid. In some embodiments, the composition is administered topically or parenterally. In some embodiments, the composition is administered via intravenous or subcutaneous injection.EXAMPLESExample 1. Phase Solubility Study

[0242] The solubility of 6S-N5-Methyl Tetrahydrofolate Calcium Salt (levomefolic acid calcium salt or LMFCa) was determined in water and several CAPTISOL® (a sulfobutylether-ß-cyclodextrin sodium) solutions at room temperature. The concentrations of CAPTISOL® were 0, 50, 100, 200, 300, and 400 mg / mL. Two milliliters of the CAPTISOL® solutions were placed in a 4 dram (15 ml) bottle with excess amount of the LMFCa solid added to the bottle. The bottle lid was tightly closed and the bottle was rotated to equilibrate overnight. Samples of the saturated solutions were taken and placed in a plastic centrifuge tube, and centrifuged 4 minutes at 3,000 rpm. Samples from the clear solution were taken and diluted in a 100mL volumetric with PBS pH 7.5 buffer, and the UV absorbance read at 290nm. The phase solubility measurement result is shown in Fig. 1. The results show that CAPTISOL® causes a linear enhancement in LMFCa solubility.Example 2. Phase Solubility Study

[0243] The solubility of folinic acid or a salt thereof is determined in water and several CAPTISOL® (a sulfobutylether-B-cyclodextrin sodium) solutions at room temperature. The concentrations of CAPTISOL® are 0, 50, 100, 200, 300, and 400 mg / mL. Two milliliters of the CAPTISOL® solutions are placed in a 4 dram (15 ml) bottle with excess amount of the LMFCa solid added to the bottle. The bottle lid is tightly closed and the bottle is rotated to equilibrate overnight. Samples of the saturated solutions are taken and placed in a plastic centrifuge tube, and centrifuged 4 minutes at 3,000 rpm. Samples from the clear solution are taken and diluted in a 100mL volumetric with PBS pH 7.5 buffer, and the UV absorbance read at 290nm.Example 3, Sample Preparation

[0244] Tablet formulation samples were prepared using 6S-N5-Methyl Tetrahydrofolate Calcium Salt (Cerbios-Pharma S A ), CAPTISOL® (CyDex Pharmaceuticals, Inc.), Avicel PH 101 NF (FMC corporation), Klucel (hydroxypropyl cellulose) (GF Pharma from Hercules), Lactose Fast Flo # 316 (Formost Food), Ac-Di-Sol (croscarmellose sodium NF) (FMC corporation), magnesium stearate NF (Mallinckrodt), and purified water.

[0245] Five formulations were prepared using the procedures below. Formulation 1 was prepared from LFMCa alone, Formulation 2 was prepared from co-grinding a mixture of levomefolic Ca salt and CAPTISOL®, Formulation 3 was prepared from trituration of a mixture of LMFCa and CAPTISOL®, Formulation 4 was prepared from freeze-drying a solution of levomefolic Ca salt and CAPTISOL®, and Formulations 5 and 6 were prepared from two different spray-drying runs from a solution of LMFCa salt and CAPTISOL®. The five formulations were made into 100 mg tablets each containing 0.5mg LMFCa salt.

[0246] A Virtis Maestro freeze-dryer was used to prepare Formulation 4, A Buchi 190 mini spray drier was used to prepare Formulations 5 and 6. The tablets were made using a Stokes Model F single punch tablet machine (with instrumentation of lower punch force measurement and a Varian VK200 tablet hardness tester).

[0247] Table 1. Ingredients in Formulation 1 using neat drugIngredient AmountLMFCa 500 mg Ac-Di-Sol 1 gramKlucel GF 2 gramMagnesium Stearate 0.5 gramAvicel PHI 01 & Lactose 96.5 gram

[0248] All ingredients (except magnesium stearate) in Formulation 1 were mixed in a bottle containing baffles and mixed on a roller mill with end over end mixing by hand over 15 minutes. The magnesium stearate was added and mixed for 2 minutes. The material was tableted on the Stokes Press at 1 tablet per second and compression force of A ton using ’Ainch standard concave round punches. The weight, thickness and hardness were measured on the batch of about 600 tablets.

[0249] Table 2. Ingredients in Formulation 2 using a Co-Ground Physical mixture of neat drug and CAPTISOL®Ingredient Amount LMFCa Salt 500 mgCAPTISOL® 2.58 mgAc-Di-Sol 1 gramKlucel GF 2 gramMagnesium Stearate 0.5 gramAvicel PHI 01 & Lactose 93.42 gram

[0250] The neat drug and CAPTISOL® was ground in a mortar to produce a drug and CAPTISOL® mixture. The same procedure was used to prepare tablets as described for Formulation 1.

[0251] Table 3, Ingredients in Formulation 3 using a Triturate mixture of neat drug and CAPTISOL® solutionIngredient Amount Drug and CAPTISOL® Triturate 3.52 mgAc-Di-Sol 1 gramKlucel GF 2 gramMagnesium Stearate 0.5 gramAvicel PH101 & Lactose 92.98 gram

[0252] A mixture of 1.16 g drug and 5.36 g of CAPTISOL® was triturated with ImL water in a mortar and dried in a vacuum oven at 45°C overnight. The mixture was ground in the mortar and assayed for drug using UV spectroscopy and found to be 14.2% drug. A sample at 3.52g would contain 500mg drug. The same procedure was used to prepare tablets as described for Formulation 1.

[0253] Table 4. Ingredients in Formulation 4 using Freeze-dried neat drug and CAPTISOL®.Ingredient AmountDrug and CAPTISOL® Freeze-dried 3.27 mgAc-Di-Sol 1 gramKlucel GF 2 gramMagnesium Stearate 0.5 gramAvicel PHI 01 & Lactose 93.23 gram

[0254] A mixture of 38.6 g drug and 173 g of CAPTISOL® was mixed and water was added to a volume of 800mL. The solution was sonicated to bring the drug into solution. The solution was freeze-dried to yield 207 g a dry yellow cake (i.e. a preformed complex mixture) which was forced through a 40-mesh stainless steel screen to fracture the cake into free-flowing plate-like shards. UV assay showed the material to be 18.2% drug. A sample of this material weighing 3.27 g would contain 500 mg LMFCa.

[0255] Table 5. Ingredients in Formulations 5 and 6 using Spray-dried neat drug and CAPTISOL®Ingredient AmountDrug and C APTISOL® Freeze-dried 4.24 mgAc-Di-Sol 1 gramKlucel GF 2 gramMagnesium Stearate 0.5 gramAvicel PHI 01 & Lactose 92.26 gram

[0256] A mixture of 2.43 g drug and 17 g of CAPTISOL® was mixed and water was added to a volume of 50 mL. The solution was sonicated to bring the drug into solution (the calculated concentration was 220 mg / mL CAPTISOL® and 42 mg / mL LMFCa). The solution was spray-dried to yield 13 g of light yellow very fine powder. UV assay showed the material to be 11.8% drug. A sample at 4.24 g would contain 500 mg drug. The two formulations using a spray-dried preparation of LMFCa and Captisol, Formulation 5 and Formulation 6, were prepared from spray-dried mixtures using similar procedure except that the spray-dried material used in Formulation 6 was dried at a slower rate than Formulation 5.Tablets were prepared using the procedure used to prepare the tablets as described for Formulation 1.Table 6. Data Collected on the Five Formulations of Tablets Prepared#4 #5Tablet #1 Neat #2 Mixture #3 TrituratedFreeze-dry Spray-dry Weight (mg) 100.6 100.9 99.2 99.1 101.6 Thickness (mm) 3.15 3.16 3.15 3.14 3.19Hardness (Kg) 15.3 16.3 13.7 15.3 15.9

[0257] The RSD for tablet weight and thickness were less than 1 % and less than 5% for hardness.Example 4, Dissolution Study

[0258] The dissolution of each tablet formulation described in Example 2 was tested. Samples from the dissolved solution were measured using the UV absorbance spectrum at 290 nm to determine the relative amount of LMFCa released from the tablets as a function of time. The dissolution study results are shown in FIG. 2 and FIG. 3.

[0259] FIG. 2 shows the dissolution curve of six tablet formulations using 900 ml simulated gastric fluid (SGF) as medium, at 37 C and 50 rpm. FIG. 3 shows the dissolution curve of six tablet formulations using 900 ml simulated intestinal fluid (SIF) as medium, at 37 °C and 50 rpm.

[0260] The disintegration time for each of the five formulation tablets are listed in Table 7.Table 7. Disintegration time for five tested formulationsFormulation # Disintegration time ( second)1 30 to 402 100 to 120 3 40 to 504 60 to 805290 to 310Example 5, Stability Study -- 3 Day test

[0261] Samples of the neat material, freeze-dried and spray-dried LMFCa + Captisol powders were placed in bottles closed tightly and bottles where the lid was left loose and placed in a stability chamber at 40°C and 75% relative humidity. After three days, thesamples were then removed from the stability oven. The samples from each bottle were weighed and dissolved in water, and assayed using HPLC to determine the peak area percent attributed to drug and other impurities including degradation products. The tables below show the calculated peak area percent attributed to only the LMFCa salt based on HPLC analysis and the difference between the peak area percent after 3 days and initially after preparation.Table 8. Comparison of T=0 and 3 Days at 40°C / 75% RH in Closed BottleChange Sample Initial (%) Final (%) (Final minus Initial) (%) LMFCa StartingMaterial 98.15 96.36 - 1.79 (No CAPTISOL®)Freeze DriedLMFCa +96.62 96.18 - 0.44 CAPTISOLSpray-driedLMFCa + 94.86 91.76 - 3.10CAPTISOLTable 9. Comparison of T=0 and 3 Days at 40°C / 75%RH in Open BottleChange (Final minus Sample Initial (%) Final (%)Initial) (%) LMFCa StartingMaterial 98.15 94.10 -4.05(No CAPTISOL®)Freeze Dried LMFCa +CAPTISOL 96.62 93.82 -2.80 Spray-dried LMFCa +CAPTISOL 94.86 91.84 -3.02

[0262] These results show that the freeze-dried samples, particularly the sample stored in the sealed bottle, had the least amount of degradation after 3 days when compared with the neat drug and spray-dried powder samples tested at the same condition.Example 6, Stability Study - 10 Day test

[0263] Samples of the neat material, freeze-dried and spray-dried LMFCa + Captisol powders were placed in bottles closed tightly and bottles where the lid was left loose and placed in a stability chamber at 40 °C and 75% relative humidity. After 10 days, the samples were then removed. The samples from each bottle were weighed and dissolved in water, and assayed using HPLC to determine the peak area percent attributed to drug and other impurities including degradation products. Table 10 below shows the calculated relative peak area percent remaining of the LMFCa after the 10-day test period based on HPLC analysis.Table 10. Recovery of LMFCa over 10 Days at 40°C / 75%RHSample t = 0 (%) 10 Days (% remaining)LMFCa Starting 100 Closed 95.4Material(No CAPTISOL®)LMFCa Starting 100 Open 89.7Material(No CAPTISOL®)Freeze Dried 100 Closed 98.6LMFCa +CAPTISOLFreeze Dried LMFCa 100 Open 95.8+ CAPTISOLSpray-dried LMFCa 100 Closed 91.9+ CAPTISOLSpray-dried LMFCa 100 Open 90.7+ CAPTISOL

[0264] Closed samples were assumed to not have picked up water.

[0265] These results show that the freeze-dried samples, particularly the sample stored in the sealed bottle, retained higher peak area percent (purity) after 10 days when compared with the neat drug and spray-dried samples tested under the same condition.Example 7. Levo Leucovorin and LMFCa Solubility Study

[0266] 50 mg of Levo Leucovorin Ca or LMFCa was weighed and placed in 3 x 4 mL glass vials. 1 mL of the indicated solvent was added to a vial, which was vortexed for 10 seconds and sonicated for 10 minutes. The vials were placed on a roller mixer overnight, then the contents were transferred to microcentrifuge tubes and subjected to centrifuge at 21,400 G at 22 °C for 5 min. The supernatant was filtered through a PVDF syringe filter. The resulting solids were diluted with water and analyzed by RP-HPLC / UV / MS The results are presented in Table 11.Table 11. Results of Levo Leucovorin and LMFCa Solubility StudyLevo Leucovorin Ca LMFCa Solventmg / mL % PA pH mg / mL % PA pHWater 32.2 97.6 7.91 9.8 60 7.5920% Captisol in water 80.4 97.2 7.80 50.5 84 8.2840% Captisol in water 85.4 95.6 7.66 68.5 88 8.0720% Captisol in Tris buffer pH 7 87.4 99.5 7.29 NT NT NT20% Captisol in Ammonium 72.5 97.8 7.68 NT NT NT Acetate buffer pH 720% Captisol in Citrate buffer NT NT NT 37.8 87.7 5.74 pH 3NT = Not TestedExample 8. Levo Leucovorin Ca and LMFCa Stability Studies

[0267] 50 mg of the indicated solid was weight into a 10 mL volumetric flask. Either buffer of 20% Captisol in buffer was added, and the flasks were vortexed for 10 seconds,followed by sonication at 50 °C for 10 minutes. Samples were collected and subjected to 1 / 50 dilution in water prior to pH measurement. The results are presented in Tables 12 and 13.Table 12. Results of Levo Leucovorin Ca Stability Study% Initial Buffer 20% Captisol pH Time (hr) 5 °C 25 °C 40 °C 5 °C 25 °C 40°C PH 2 0 100.00 100.00 100.00 100.00 100.00 100.003 44.14 84.20 70.73 93.81 75.88 42.44 24 37.81 45.11 17.65 79.61 24.65 12.30 48 37.58 24.39 11.58 65.75 14.07 11.47 312 44.40 7.26 5.18 23.32 10.35 10.20 pH 4.5 0.00 100.00 100.00 100.00 100.00 100.00 100.003.00 98.99 98.71 96.45 99.57 101.00 102.79 6.00 98.93 97.93 96.09 104.42 102.88 101.28 24.00 98.84 93.23 81.50 103.16 102.03 96.52 48.00 97.96 86.97 70.99 105.57 93.49 91.71 168.00 93.91 65.80 42.47 101.87 90.85 65.71 pH 7 0 100.00 100.00 100.00 100.00 100.00 100.00 TRIS 24 100.48 101.19 101.26 102.13 103.13 101.2348 99.99 100.78 101.52 98.43 100.76 100.69 312 101.26 101.43 98.79 100.95 104.48 100.71 pH 7 0 100.00 100.00 100.00 100.00 100.00 100.00 Am. Ac. 24 73.64 98.77 98.25 98.29 97.75 99.5148 98.93 98.26 98.09 97.69 98.21 97.73 312 99.15 93.30 94.65 103.00 100.67 100.32Table 13, Results of LMFCa Stability Study% Initial Buffer 20% Captisol pH Time (hr) 5 °C 25 °C 40 °C 5 °C 25 °C 40°C pH 2 0 100.00 100.00 100.00 100.00 100.00 100.003 91.06 76.94 80.75 102.44 100.53 99.17 24 52.63 34.27 39.58 103.15 96.77 95.00 48 24.90 23.47 28.75 98.03 92.37 84.06 312 60.09 13.36 4.30 92.78 63.68 27.18 pH 4.5 0.00 100.00 100.00 100.00 100.00 100.00 100.003.00 57.30 59.12 46.37 94.46 95.52 100.58 6.00 42.26 40.17 33.55 99.18 101.65 97.21 24.00 13.10 6.89 5.53 98.41 94.37 85.11 48.00 5.40 3.49 8.80 98.54 88.61 75.01 168.00 1.54 9.80 10.96 92.39 58.29 42.73 pH 7 0 100.00 100.00 100.00 100.00 100.00 100.00 TRIS 24 95.76 80.92 65.37 95.31 91.66 82.3548 91.55 64.37 41.22 97.24 85.98 74.52 312 72.94 10.75 16.84 86.29 51.00 16.49 pH 7 0 100.00 100.00 100.00 100.00 100.00 100.00 Am. Ac. 24 93.70 80.30 67.02 95.48 91.72 83.7748 89.85 65.59 44.17 97.06 84.04 76.49 312 73.24 9.74 13.37 89.77 51.56 11.84Example 9, Levo Leucovorin Oral Solution Stability Study

[0268] Formulations la through 4a were prepared as follows. Formulation la was prepared by weighing Levo Leucovorin Ca and transferring into a 25 mL volumetric flask. Nanopure water was added almost to the fill line, visibly ensuring that the drug was dissolved. Concentrated NaOH or HCl was added while using a calibrated pH meter to bring the pH to 7.8. Additional nanopure water was added to bring the solution to the fill line. Aliquots of thesolution were added to 4 mL glass vials subsequently sealed with screw caps. In a taste test, Formulation la had no taste.Table 14, Formulation la: 10 mg / mL Drug, No Tris, No Captisol, pH 7,8Concentration mg for each mg for each (mg / mL) 100mL 25mL Levo Leucovorin Ca (1010 1,000 250 mg / mL)HCl / NaOH adj pH 7.8* as needed as needed as neededCaptisol (0%) 0 0 0*pH 7.8 target (acceptable range 6.5-8.5)

[0269] Formulations 2a and 3a were prepared by weighing Levo Leucovorin Ca and transferring into a 25 mL volumetric flask. A solution of Captisol in nanopure water was prepared separately and added to the volumetric flask almost to the fill line, visibly ensuring that the drug is dissolved. Concentrated NaOH or HCl was added while using a calibrated pH meter to bring the pH to 7.8. Additional nanopure water was added to bring the solution to the fill line. Aliquots of the solution were added to 4 mL glass vials subsequently sealed with screw caps. In a taste test, Formulation 2a had a faint taste of salt and Formulation 3a had a slight taste of salt.Table 15, Formulation 2a: 10 mg / mL Drug, No Tris, 10% Captisol, pH 7.8Captisol weight adjusted for Concentration mg for each mg for each 5.0% H2O content (mg / mL) lOOmL 25mL Levo Leucovorin Ca (1010 1,000 250 mg / mL)HCl / NaOH adj pH 7.8* 0 0 0 Captisol (10%, H2O100 10,526.3 2,631.6corrected)*pH 7.8 target (acceptable range 6.5-8.5)Table 16. Formulation 3a: 10 mg / mL Drug, No Tris, 20% Captisol, pH 7.8Captisol weight adjusted for Concentration mg for each mg for each 5.0% H2O content (mg / mL) 100mL 25mL Levo Leucovorin Ca (1010 1,000 250 mg / mL)HCl / NaOH adj pH 7.8* 0 0 0 Captisol (20%, H2O200 21,052.6 5,263.2corrected)*pH 7.8 target (acceptable range 6.5-8.5)Formulation 4a was prepared by weighing Levo Leucovorin Ca and transferring into a 25 mL volumetric flask. A Captisol / Tris buffer solution in nanopure water was prepared separately and added to the volumetric flask almost to the fill line, visibly ensuring that the drug is dissolved. Concentrated NaOH or HCl was added while using a calibrated pH meter to bring the pH to 7.8. Additional nanopure water was added to bring the solution to the fill line. Aliquots of the solution were added to 4 mL glass vials subsequently sealed with screw caps. In a taste test, Formulation 4a had a slight taste of salt.Table 17. Formulation 4a: 10 mg / mL Drug, 50 mM Tris, 20% Captisol, pH 7.8 Captisol weight adjusted for Concentration mg for each mg for each 5.0% H2O content (mg / mL) 100mL 25mL Levo Leucovorin Ca (1010 1,000 250 mg / mL)Tris Buffer, Base (50 mM2.148 214.8 53.7 final)Tris Buffer, HCl (50 mM5.084 508.4 127.1 final)Captisol (20%, H2O200 21,052.6 5,263.2corrected)*pH 7.8 target (acceptable range 6.5-8.5)

[0270] The vials from each formulation were placed in stability chambers creating one of three sets of storage conditions: 5 °C, 25 °C and 60% relative humidity, or 40 °C and 75% relative humidity. A sample from each of the vials was removed at six days and at two weeks, and samples are removed at one month, three months, and six months. The samples from each vial were removed by pipette and diluted in water and assayed using HPLC todetermine the peak area or peak area % attributed to drug and other impurities including degradation products. Six days 5 °C is used as TO for all formulations.Table 18. Formulations la-4a - Assay 5 °C (Target 10 mg / mL)Time Point TO 6 days 2 weeksFormulation la 9.93 9.93 9.77Formulation 2a 9.81 9.81 9.84Formulation 3a 9.90 9.90 9.84Formulation 4a 9.95 9.95 9.46Table 19. Formulations 1a-4a – Assay 25 °C / 60% RH (Target 10 mg / mL)Time Point TO 6 days 2 weeksFormulation la 9.93 9.92 9.79Formulation 2a 9.81 9.77 9.68Formulation 3a 9.90 9.89 9.89Formulation 4a 9.95 9.91 9.72Table 20. Formulations la-4a - Assay 40 °C / 75% RH (Target 10 mg / mL)Time Point TO 6 days 2 weeksFormulation la 9.93 9.89 9.65Formulation 2a 9.81 9.92 9.66Formulation 3a 9.90 9.90 9.57Formulation 4a 9.95 9.91 9.62

[0271] The pH of each sample is provided in the tables below. Six days 5 °C is used as TO for all formulations.Table 21, Formulations la-4a - Assay 5 °C (Target 10 nig / mL)Time Point TO 6 days 2 weeksFormulation la 7.89 8.03 7.81Formulation 2a 7.64 7.78 7.63Formulation 3a 7.68 7.59 7.46Formulation 4a 7.93 7.96 7.97Table 22, Formulations la-4a ~ Assay 25 °C / 60% RH (Target 10 nig / mL)Time Point TO 6 days 2 weeksFormulation la 7.89 8.12 8.04Formulation 2a 7.64 7.81 7.43Formulation 3a 7.68 7.56 7.46Formulation 4a 7.93 7.92 7.93Table 23, Formulations la-4a - Assay 40 °C / 75% RH (Target 10 mg / mL)Time Point TO 6 days 2 weeksFormulation la 7.89 7.95 7.60Formulation 2a 7.64 7.48 7.55Formulation 3a 7.68 7.33 7.23Formulation 4a 7.93 7.91 7.91

[0272] The appearance of each sample is provided in the tables below. Six days 5 °C is used as TO for all formulations.Table 24, Formulations la-4a - Assay 5 °C (Target 10 mg / mL)Time Point TO 6 daysFormulation la Clear ClearFormulation 2a Clear ClearFormulation 3a Clear ClearFormulation 4a Clear ClearTable 25, Formulations la-4a ~ Assay 25 °C / 60% RH (Target 10 mg / mL)Time Point TO 6 daysFormulation la Clear ClearFormulation 2a Clear ClearFormulation 3a Clear ClearFormulation 4a Clear ClearTable 26, Formulations la-4a - Assay 40 °C / 75% RH (Target 10 mg / mL)Time Point TO 6 daysVery' slightFormulation la ClearyellowVery slightFormulation 2a ClearyellowVery slightFormulation 3a ClearyellowVery slightFormulation 4a ClearyellowExample 10. Levo Leucovorin Powder for Oral Solution Stability Study

[0273] Formulations 5a through 9a were prepared as follows. Formulations 5a, 6a, and 7a were prepared by weighing the indicated amounts of methyl paraben, potassium sorbate, xylitol, and strawberry flavor to make a pre-blend appropriate for a 25 mL batch. The ingredients were mixed with a mortar and pestle. The requisite amount of Levo Leucovorin Ca was weighed and transferred into a 4 mL glass vial. The requisite amount of pre-blend wasweighed and added to the vial. If called for, the requisite amount of Captisol was weighed and added to the vial. The vial was sealed with a screw cap and the contents were mixed in the vial. The entries in the Concentration column in Tables 27 through 31 indicate the concentration of that substance when a sample of the formulation is reconstituted in water. In a taste test, Formulation 5a had a faint taste of strawberry, Formulation 6a had a taste of strawberry with a faint taste of salt, and Formulation 7a had a faint taste of strawberry' with a taste of salt.Table 27. Formulation 5a CompositionConcentration mg for each mg for eachComponent Amount (%)(mg / mL) lOOmL 25mLLevo Leucovorin Ca (1010 1,000.0 250.0 34.2 mg / mL)Methyl Paraben 0.8 80.0 20.0 2.7 Potassium Sorbate 2.1 210.0 52.5 7.2 Xylitol 14 1,400.0 350.0 47.9Strawberry Flavor 2.3 230.0 57.5 7.9 Table 28. Formulation 6a CompositionConcentration mg for each mg for eachComponent Amount (%)(mg / mL) 1 OOmL 25mLLevo Leucovorin Ca (1010 1,000.0 250.0 7.7 mg / mL)Captisol (10%)* 100 10,526.3 2631.6 77.4 Methyl Paraben 0.8 80.0 20.0 0.6 Potassium Sorbate 2.1 210.0 52.5 1.6 Xylitol 14 1,400.0 350.0 10.8Strawberry Flavor 23 230.0 57.5 1.8 * Captisol weight adju ted for 5.0% H2O contentTable 29. Formulation 7a CompositionConcentration mg for each mg for eachComponent Amount (%)(mg / mL) 1 OOmL 25mLLevo Leucovorin Ca (1010 1,000.0 250.0 4.4 mg / mL)Captisol (20%)* 200 21,052.6 5,263.2 87.3 Methyl Paraben 0.8 80.0 20.0 0.3 Potassium Sorbate 2.1 210.0 52.5 0.9 Xylitol 14 1,400.0 350.0 6.1Strawberry Flavor 2.3 230.0 57.5 1.0*Captisol weight adjusted for 5.0% H2O content

[0274] Formulation 8a was prepared by weighing the indicated amounts of methyl paraben, potassium sorbate, xylitol, strawberry flavor, and Tris buffer components to make a pre-blend appropriate for a 25 mL batch. The ingredients were mixed with a mortar and pestle. The requisite amount of Levo Leucovorin Ca was weighed and transferred into a 4 mL glass vial. The requisite amount of pre-blend was weighed and added to the vial. The requisite amount of Captisol was weighed and added to the vial. The vial was sealed with a screw cap and the contents were mixed in the vial. In a taste test, Formulation 8a had a faint taste of strawberry with a taste of salt.Table 30. Formulation 8a CompositionConcentration mg for each mg for eachComponent Amount (%)(mg / mL) lOOmL 25mLLevo Leucovorin Ca (1010 1,000.0 250.0 4.23 mg / mL)Tris Buffer, Base (50 mM2.148 214.8 53.7 0.91 final)Tris Buffer, HCl (50 mM5.084 508.4 127.1 2.15 final)Captisol (20%)* 200 21,052.6 5,263.2 84.59 Methyl Paraben 0.8 80.0 20.0 0.34 Potassium Sorbate 2.1 210.0 52.5 0.89 Xylitol 14 1,400.0 350.0 5.92Strawberry Flavor 2.3 230.0 57.5 0.97 *Captisol weight adjusted for 5.0% H2O content

[0275] Formulation 9a was prepared by weighing the indicated amounts of methyl paraben, potassium sorbate, sucralose, strawberry flavor, and Tris buffer components to make a pre-blend appropriate for a 25 mL batch. The ingredients were mixed with a mortar and pestle. The requ isite amount of Levo Leucovorin Ca was weighed and transferred into a 4 mL glass vial. The requisite amount of pre-blend was weighed and added to the vial. The requisite amount of Captisol was weighed and added to the vial. The vial was sealed with a screw cap and the contents were mixed in the vial. In a taste test. Formulation 9a had a very sweet taste with a faint taste of strawberry.Table 31, Formulation 9a: 10 mg / mL Drug, 50mM Tris, 20% Captisol, Sucralose Concentration mg for each mg for eachComponent Amount (%)(mg / mL) lOOmL 25mLLevo Leucovorin Ca (1010 1,000.0 250.0 4.23 mg / mL)Tris Buffer, Base (50 mM2.148 214.8 53.7 0.91 final)Tris Buffer, HCl 50 mM5.084 508.4 127.1 2.15 final)Captisol (20%)* 200 21,052.6 5,263.2 84.59 Methyl Paraben 0.8 80.0 20.0 0.34 Potassium Sorbate 2.1 210.0 52.5 0.89 Sucralose 14 1,400.0 350.0 5.92Strawberry Flavor 2.3 230.0 57.5 0.97 * Captisol weight adj sted for 5.0% H2O content

[0276] Samples of Formulations 5a - 9a in 4 mL glass vials were placed in a stability chamber. For each of Formulations 5a through 9a, at least 10 vials were prepared. The three vials were stored in the chamber at 5 °C. Three vials were stored at 25 °C and 60% relative humidity. Three vials were stored at 40 °C and 75% relative humidity. A vial will be removed at one month, three months, and six months for analysis. After allowing each vial to come to room temperature, water is added to each vial to prepare 2 mL of the reconstituted oral solution formulation. The following amounts of water are added to each formulation: 1.99, 1.86, 1.73, 1.70, 1.70 mL for Formulations 5a, 6a, 7a, 8a, 9a, respectively. A sample from each vial is removed by pipette and diluted in water and assayed using HPLC to determine the peak area or peak area % attributed to drug and other impurities including degradation products. Data collected from samples that were collected at To for the formulations are shown in the following table.Table 32. Formulations 5a-9a at TOpH Appearance Time for ConstitutionWhite / Off-whi te Invert / vortex 2.5 min then Formulation 5a 7.13Powder sonicate for 2 x 10 minWhi te / Off-white Invert / vortex 2.5 min then Formulation 6a 7.28Powder sonicate for 2 x 10 minWhite / Off-white Invert / vortex 2.5 min then Formulation 7a 7.21Powder sonicate for 2 x 10 minWhite / Off-white Invert / vortex 2.5 min then Formulation 8a 8.01Powder sonicate for 2 x 10 minWhi te / Off-white Invert / vortex 2.5 min then Formulation 9a 8.02Powder sonicate for 2 x 10 minExample 11. Levo Leucovorin Excipient Compatibility Study

[0277] Samples of Levo Leucovorin Calcium (10 mg) and an excipient shown in the following table were weighed and place into 4x 8 mL glass vials. The vials were subjected to vortex and then tapped to settle the powder. Three of the vials were placed in an oven at 60 °C, leaving one vial out of the oven for TO analysis. At each time point (0, 1 week, 2 weeks, and 4 weeks) a vial was removed from the oven (if it had been in the oven), brought to room temperature, combined with 5 mL of nanopure water, and subjected to vortex until the solution turned clear. The solution was then diluted a further 1 / 20 into autosampler vials with nanopure water for analysis. A Levo Leucovorin Ca standard at 0.1 mg / mL was prepared by weighing 1 mg into a 5 mL volumetric flask and dissolving it in nanopure water. This was diluted 1 / 2 into an autosampler vial in water. The samples and standard were analyzed by UPLC / UV. After 1 week analysis, the sample with sucralose was discolored (slightly pinkish), so additional vials were put on stability of the excipients alone at the same concentrations as the excipient + L-Leucovorin Ca.Table 33, Excipient Concentrations in the Compatibility StudyExcipient AmountSucralose 14 nigStrawberry flavor 2.3 mgPotassium sorbate 2.1 mgMethyl paraben 0.8 mgXylitol 14 mgGrape flavor 2.3 mgBerry flavor 2.3 mg

[0278] Results: all of the excipients except sucralose remained stable and did not alter the L-Leucovorin Ca stability over 4 weeks at 60 °C.

Claims

WHAT IS CLAIMED IS:1, A pharmaceutical composition comprising:(iii) a compound having the structure of Formula (II):(II)or a pharmaceutically acceptable salt thereof, tautomer thereof, or ionic form thereof, wherein:each of RA, RB, and RCis independently selected from -H, -Ci-6 alkyl, -C2-ioalkenyl, -C2-ioalkynyl, -C(O)H, and -C(O)Ci-6alkyl, orRBand RCtogether with intervening atoms form a 4-8 membered ring, optionally comprising one or more double bond; and(iv) a cyclodextrin.

2. The pharmaceutical composition of claim 1, wherein the ionic form is a cationic form.

3. The pharmaceutical composition of claim 1 or 2, wherein the compound of Formula (II) is compound, or pharmaceutically acceptable salt thereof, tautomer thereof, or cationic form thereof, having the structure of Formula (Il-a):(Il-a).

4. The pharmaceutical composition of claim 1 or 2, wherein the compound of Formula (II) is compound, or pharmaceutically acceptable salt thereof, tautomer thereof, or cationic form thereof, having the structure of Formula (Il-b):(Il-b).

5. The pharm ceutical composition of claim 1 or 2, wherein the compound of Formula (II) is compound, or pharmaceutically acceptable salt thereof, tautomer thereof, or cationic form thereof, ha ving the structure of Formula (II-c):

6. The pharmaceutical composition of any one of claims 1 to 5, wherein RAis -H.

7. The pharmaceutical composition of any one of claims 1 to 6, wherein RBis -II, -CH3, or -C(O)H.

8. The pharmaceutical composition of any one of claims 1 to 7, wherein RCis -H, -CH3, or -C(O)H.

9. The pharmaceutical composition of any one of claims 1 to 5, wherein RBand RCtogether with intervening atoms form a five membered ring,10. The pharmaceutical composition of claim 1, wherein the compound of Formula (II), or pharmaceutically acceptable salt thereof, tautomer thereof, or cationic form thereof is selected from the group consisting of:

11. The pharmaceutical composition of claim 1, wherein the compound of Formula (II), or pharmaceutically acceptable salt thereof, tautomer thereof, or cationic form thereof is selected from the group co sisting of.CK ^OH CK ^0H12. The pharmaceutical composition of claim I, wherein the compound of Formula (II) isor a pharmaceutically acceptable salt thereof, tautomer thereof, or cationic form thereof.

13. The pharmaceutical composition of claim 1, wherein the compound of Formula (II) isor a pharmaceutically acceptable salt thereof, tautomer thereof, or cationic form thereof.

14. The pharmaceutical composition of any one of claims 1 to 13, wherein the pharmaceutically acceptable salt of the compound of Formula (II) is a sodium, potassium, calcium, or magnesium salt.

15. The pharmaceutical composition of claim 14, wherein the pharmaceutically acceptable salt of the compound of Formula (II) is a calcium salt.

16. The pharmaceutical composition of any one of claims 1 to 15, wherein the cyclodextrin is a sulfoalkyl ether cyclodextrin.

17. The pharmaceutical composition of any one of claims 1 to 16, wherein the cyclodextrin is a compound, or a mixture thereof, of Formula (I):or a pharmaceutically acceptable salt thereof, wherein:p is 4, 5, or 6;each Ri is independently selected from -OH and -O-(Ci-Cs alkylene -SOsT; andT is selected from hydrogen or a pharmaceutically acceptable cation thereof; provided that at least one R1is -O-(C1-C8alkylene)-SO3T.

18. The pharmaceutical composition of claim 17, wherein at least one Ri is OCH2CH2CH2CH2SO3T.

19. The pharmaceutical composition of any one of claims 17 to 18, wherein the pharmaceutically acceptable cation is sodium or potassium.

20. The pharmaceutical composition of cl im 17 or 18, wherein the pharmaceutically acceptable cation is a cation of a (Ci-C6)-alkanolamine.

21. The pharmaceutical composition of claim 20, wherein the alkanolamine is ethanolamine, methanolamine, 2-amino-2-methyl-l -propanol, valinol, orN-methyl-(2,3,4,5,6-pentahydroxy-hexyl)-amine.

22. The pharmaceutical composition of any one of claims 17 to 18, wherein the pharm ceutically acceptable cation is a cationic form of the compound of Formula (II).

23. The pharmaceutical composition of claim 22, wherein the pharmaceutically acceptable cation is cation of the compound having the structure:

24. The pharmaceutical composition of any one of claims 1 to 23, wherein the composition further comprises a pH adjusting agent.

25. The pharmaceutical composition of claim 24, wherein the pH adjusting agent is N-methyl-(2,3,4,5,6-pentahydroxy-hexyl)-amine.

26. The pharmaceutical composition of any one of claims 1 to 25, wherein the composition does not contain clopidogrel.

27. The pharmaceutical composition of any one of claims 1 to 26, wherein the composition is in a solid form,28. The pharmaceutical composition of any one of claims 1 to 27, wherein the composition is formulated into a pill, capsule, granules, or tablet.

29. The pharmaceutical composition of any one of claims 1 to 27, wherein the composition is formulated into a reconstitutable solid.

30. The pharmaceutical composition of any one of claims 1 to 29, wherein the composition is a lyophile.

31. The pharmaceutical composition of any one of claims 1 to 26, wherein the composition is formulated into a liquid form suitable for injection.

32. The pharmaceutical composition of any one of claims 1 to 26, wherein the composition is formulated into a liquid form suitable for oral administration.

33. The composition of any one of claims I to 32, further comprising an excipient.

34. The pharmaceutical composition of any one of claims 1 to 33, wherein the composition is characterized by compound of Formula (11) being in an amount of at least 96% by weight in relation to an initial content of the compound of Formula (II) after storage in a closed container for 10 days at 40 °C and 75% relative humidity.

35. The pharmaceutical composition of any one of claims 1 to 34, further comprising a chemotherapeutic agent.

36. The pharmaceutical composition of claim 35, wherein the chemotherapeutic agent is selected from the group consisting of: methotrexate, pyrimethamine, pralatrexate, 5-fluorouracil, pyrimethamine, doxorubicin, epirubicin, idarubicin, daunorubicin, and cyclophosphamide.

37. A method of preparing the composition of any one of claims 1 to 36, comprising:providing a solution containing the compound of Formula (II), or a pharmaceutically acceptable salt thereof, tautomer thereof, or ionic form thereof, and cyclodextrin; andfreeze-drying the solution.

38. A method of preparing the composition of any one of claims 1 to 26, comprising:providing a dry powder containing the compound of Formula (II), or a pharmaceutically acceptable salt thereof, tautomer thereof, or ionic form thereof, and a cyclodextrin; andreconstituting the dry powder into a liquid.

39. A method of reducing an oxidative degradation of a compound of Formula (II), comprising combining an amount of the compound of Formula (II) and an amount of cyclodextrin, wherein the amount of cyclodextrin is effective in reducing the oxidative degradation of the compound of Formula (II).

40. The method of claim 39, wherein the molar ratio of the cyclodextrin to the folic acid or folic acid derivative is in the range of about 0.1 to about 10.

41. The method of claim 40, wherein the molar ratio is about 1.0 to about 1.

2.

42. A method of reducing a female subj ect’ s risk of having a miscarriage or having a fetus with a birth defect, comprising administering to the female subject in need thereof an effective amount of the composition of any one of claims 1 to 34.

43. The method of claim 42, wherein the birth defect is a neural tube defect, a cleft lip defect, or a cleft palate defect.

44. A method of treating a deficiency or disease positively affected by the administration of both folates and reduced folates selected from the group consisting of subacute encephalitis associated with dementia, vacuolar myelopathy, premature occlusive arterial disease, severe vascular disease in infancy, severe vascular disease in childhood, progressive arterial stenosis, intermittent claudication, renovascular hypertension, ischemic cerebrovascular disease, premature retinal artery occlusion, premature retinal vein occlusion, cerebral occlusive arterial disease, occlusive peripheral arterial disease, premature death due to thromboembolic disease, premature death due to ischemic heart disease, psoriasis, celiac disease, arthritic condition, inflammation conditions, megaloblastic anaemia due to folate deficiency, intestinal malabsorption, and depressive illness, said method comprising administering to a subject in need thereof an effective amount of the composition of any one of claims 1 to 36.

45. A method of reducing a female's risk of having a miscarriage, reducing a female's risk of having a fetus with a neural tube defect, reducing a female's risk of having a fetus with a cleft lip defect, reducing a female’s risk of having a fetus with a cleft palate defect, said method comprising administering to the female subject in need thereof an effective amount of any one of the composition of any one of claims I to 36.

46. A method of reducing the side effects of a chemotherapeutic agent in a subject, said method comprising administering to the subject in need thereof an effective amount the composition of any one of claims 1 to 36.

47. The method of claim 46, wherein the side effects are selected from the group consisting of: megaloblastic anemia, nausea, vomiting, and combinations thereof.

48. The method of claim 46 or 47, wherein the chemotherapeutic agent is selected from the group consisting of: methotrexate, pralatrexate, 5 -fluorouracil, pyrimethamine, doxorubicin, epirubicin, idarubicin, daunorubicin, and cyclophosphamide.

49. The method of claim 48, wherein the chemotherapeutic agent is 5 -fluorouracil or methotrexate.

50. A method of treating cancer in a subject, said method comprising administering to a subject in need thereof an effective amount of the composition of any one of claims 1 to 36.

51. The method of claim 50, further comprising administering one or more chemotherapeutic agent to the subject.

52. The method of claim 51, wherein the chemotherapeutic agent is methotrexate, pralatrexate, 5 -fluorouracil, pyrimethamine, doxorubicin, epirubicin, idarubicin, daunorubicin, and cyclophosphamide.

53. A method of therapeutically treating or preventing a neurodevelopmental disorder, said method comprising administering to a subject in need thereof an effective amount of the composition of any one of claims 1 to 36.

54. The method of claim 53, wherein the neurodevelopmental disorder may be fragile X syndrome, autism spectrum disorder, attention deficit hyperactivity disorder (ADHD), pervasive developmental disorder-not otherwise specified (PDD-NOS), Rett syndrome, Asperger syndrome, Angelman syndrome, generalized anxiety disorder, social anxiety disorder, bipolar disorder, epilepsy, oppositional defiant disorder, cerebral folate deficiency, FOLRl-related cerebral folate transport deficiency, or a sleep disorder.

55. The method of any one of claims 53 to 54, wherein the administration reduces symptoms associated with neurodevelopment disorders in said subject.

56. The method of claim 55, wherein symptoms associated with neurodevelopment disorder may be selected from the group consisting of irritability, homing deficits, deficits inadaptive functioning, chronic pain, seizures, difficulty with communication, difficulty with social interactions, obsessive interests, repetitive behaviors, inappropriate social interaction, poor eye contact, compulsive behavior, impulsivity, repetitive movements, self-harm, persistent repetition of words or actions learning disabilities, speech delays, depression, anxiety, changes, tics, and sound sensitivity.

57. The method of any one of claims 53 to 56, wherein the neurodevelopmental disorder is autism spectrum disorder is ASD.

58. The method of claim 57, wherein said method results in a reduction of the symptoms of ASD that is characterized by improvement according to the Vineland Adaptive Behavior Scales, ADOS-2, CGI-C social deficit subscales, aberrant behavior checklist, the Autism Treatment Evaluation Checklist (ATEC), Social Responsiveness Scale, 2ndEdition (SRS-2), or any combination thereof.

59. A method of reducing a female's risk of having a fetus with a neurodevelopmental disorder, said method comprising administering to the female in need thereof an effective amount of a composition of any one of claims 1 to 36.

60. The method of claim 59, wherein the neurodevelopmental disorder is fragile X syndrome, autism spectrum disorder, attention deficit hyperactivity disorder (ADHD), pervasive developmental disorder-not otherwise specified (PDD-NOS), Rett syndrome, Asperger syndrome, Angelman syndrome, generalized anxiety disorder, social anxiety disorder, bipolar disorder, epilepsy, oppositional defiant disorder, or a sleep disorder.

61. The method of claim 60, the neurodevelopmental disorder is autism spectrum disorder.

62. The method of any one of Claims 59 to 61, wherein said method comprises administering the composition at preconception, during the first trimester of pregnancy, during the second trimester of pregnancy, or during the third trimester of pregnancy.

63. The method of any one of claims 42 to 62, wherein the subject is a human.

64. The method of any one of claims 42 to 63, wherein the composition is administered orally.

65. The method of claim 64, where the composition is administered as a liquid.

66. The method of claim 65, comprising:providing a dry powder comprising the compound of Formula (II) and the composition;adding a diluent to the dry powder;mixing the dry powder and diluent to form a liquid formulation; and administering the liquid formulation to the subject.

67. The method of any one of claims 42 to 63, wherein the composition is administered parenterally.

68. The method of any one of claims 42 to 63, wherein the composition is administered intravenously.

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