Anticonvulsant activity of steroids

Abstract

To provide methods of treating, reducing and / or mitigating symptoms associated with and / or caused by traumatic brain injury, Alzheimer's disease, epilepsy, anxiety, fragile X syndrome, post-traumatic stress disorder, lysosomal storage disorders (Niemann-Pick type C disease), depression (including post-partum depression), premenstrual dysphoric disorder, alcohol craving, and smoking cessation in a subject in need of treating, reducing and / or mitigating the symptoms.SOLUTION: The present invention relates to methods of preventing, inhibiting, delaying, and / or mitigating seizures by administration of a steroid, e.g., a neurosteroid, e.g., allopregnanolone.SELECTED DRAWING: Figure 2

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 61 / 732,252, filed November 30, 2012, which is incorporated herein by reference in its entirety for all purposes.

[0002] Field The present invention relates to methods of preventing, suppressing, delaying and / or alleviating seizures by administration of a steroid, eg, a neurosteroid, eg, allopregnanolone. [Background technology]

[0003] background Steroids, including neurosteroids (e.g., allopregnanolone), are significantly insoluble in aqueous solutions. Various approaches are used to increase aqueous solubility, such as the use of cyclodextrin solutions. However, even with the use of cyclodextrin as a solvating aid, the solubility is still not sufficient to allow systemic delivery to treat medical conditions. Summary of the Invention

[0004] overview In one aspect, provided are methods for preventing, treating, reducing, and / or alleviating one or more symptoms associated with and / or caused by traumatic brain injury, Alzheimer's disease, epilepsy, anxiety, fragile X syndrome, post-traumatic stress disorder, lysosomal storage disorders (Niemann-Pick type C), depression (including postpartum depression), premenstrual dysphoric disorder, alcohol craving, and smoking cessation in a subject in need thereof. In some embodiments, the method comprises administering a steroid to the subject.

[0005] In another aspect, there is provided a method for preventing, treating, reducing, and / or alleviating symptoms associated with and / or caused by epilepsy in a subject in need thereof, hi some embodiments, the method comprises administering a steroid to the subject.

[0006] In yet another aspect, methods are provided for promoting the cessation or interruption of an impending seizure in a subject in need thereof, hi some embodiments, the methods comprise administering a steroid to the subject.

[0007] With respect to the embodiments of the present methods, in some embodiments, the steroid is a neurosteroid. In some embodiments, the neurosteroid is selected from the group consisting of allopregnanolone, allotetrahydrodeoxycorticosterone, ganaxolone, alphaxolone, alphadolone, hydroxydione, minaxolone, and artesin. In some embodiments, the neurosteroid is allopregnanolone. In some embodiments, the steroid is formulated in a cyclodextrin. In various embodiments, the steroid is formulated in hydroxypropyl-β-cyclodextrin or sulfobutylether-β-cyclodextrin sodium salt. In some embodiments, the subject is experiencing an aura. In some embodiments, the subject is warned of an impending seizure. In some embodiments, the subject is experiencing a seizure. In some embodiments, the subject has status epilepticus. In some embodiments, the subject has myoclonic epilepsy. In some embodiments, the subject is suffering from a cluster of seizures. In some embodiments, the seizures are tonic seizures. In some embodiments, the seizures are clonic seizures. In some embodiments, the subject is human. In some embodiments, the steroid is administered intramuscularly, intravenously, or subcutaneously. In some embodiments, the method involves treating, reducing, and / or alleviating symptoms associated with and / or caused by epilepsy by administering allopregnanolone formulated in sulfobutylether-β-cyclodextrin sodium salt intramuscularly (im), subcutaneously (sc), or intravenously (iv). In some embodiments, the epilepsy is status epilepticus. In some embodiments, the steroid or neurosteroid (e.g., allopregnanolone) is administered at a dose ranging from about 0.25 mg / kg to about 15 mg / kg, e.g., about 0.25, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 mg / kg. In some embodiments, the steroid or neurosteroid (e.g., allopregnanolone) is self-administered by the subject. In some embodiments, the steroid or neurosteroid (e.g., allopregnanolone) is administered by a caregiver who is not the subject.

[0008] In yet another aspect, a composition comprising, or consisting essentially of, a steroid and a cyclodextrin is provided. In some embodiments, the steroid is a neurosteroid. In some embodiments, the neurosteroid is selected from the group consisting of allopregnanolone, allotetrahydrodeoxycorticosterone, ganaxolone, alphaxolone, alphadolone, hydroxydione, minaxolone, and althesin. In some embodiments, the steroid is allopregnanolone. In some embodiments, the cyclodextrin is hydroxypropyl-β-cyclodextrin, sulfobutylether-β-cyclodextrin sodium salt, or a mixture thereof. In some embodiments, the composition comprises allopregnanolone and sulfobutylether-β-cyclodextrin sodium salt.

[0009] In some embodiments, the steroid or neurosteroid (e.g., allopregnanolone) is administered by inhaler or formulated for administration by inhaler. In some embodiments, the steroid or neurosteroid (e.g., allopregnanolone) is atomized or aerosolized. In some embodiments, the steroid or neurosteroid (e.g., allopregnanolone) is atomized or aerosolized without heating. In some embodiments, the atomized or aerosolized steroid or neurosteroid (e.g., allopregnanolone) particles have a mass median aerodynamic diameter ("MMAD") of about 5 μm or less. In some embodiments, the atomized or aerosolized steroid or neurosteroid (e.g., allopregnanolone) particles have a mass median aerodynamic diameter ("MMAD") of about 2-3 μm. In some embodiments, the steroid or neurosteroid (e.g., allopregnanolone) is delivered to the distal alveoli.

[0010] definition As used herein, "administering" refers to local and systemic administration, including, for example, enteral, parenteral, pulmonary, and topical / transdermal administration. Routes of administration of steroids or neurosteroids (e.g., allopregnanolone) that can be used in the methods described herein include, for example, oral (per os (PO)), nasal, inhalation, or pulmonary administration, administration as a suppository, topical contact, transdermal delivery (e.g., via a transdermal patch), intrathecal (IT), intravenous ("iv"), intraperitoneal ("ip"), intramuscular ("im"), or subcutaneous ("sc") administration to a subject, or implantation of a sustained-release device, such as a mini-osmotic pump, depot, or the like. Administration can be by any route, including parenteral and transmucosal (e.g., oral, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, for example, intravenous, intramuscular, intraarterial, intradermal, subcutaneous, intraperitoneal, intravenous, iontophoretic, and intracranial. Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, transdermal patches, etc.

[0011] The terms "systemic administration" and "systemically administered" refer to a method of administering a compound or composition to a mammal via the circulatory system so that the compound or composition is delivered to a site within the body, including the target site of pharmaceutical action. Systemic administration includes, but is not limited to, oral, intranasal, rectal, and parenteral (e.g., other than through the digestive tract, e.g., intramuscular, intravenous, intraarterial, transdermal, and subcutaneous) administration.

[0012] The term "co-administration" refers to the simultaneous presence of both active agents in the blood. Co-administered active agents can be delivered concurrently (i.e., simultaneously) or sequentially.

[0013] The term "administering" refers to the act of a medical professional (e.g., a physician) or other person managing a subject's medical care to administer and / or authorize the administration of a steroid or neurosteroid (e.g., allopregnanolone) to the subject. Administering can involve diagnosing and / or determining an appropriate therapeutic or prophylactic regimen, and / or prescribing a particular steroid or neurosteroid (e.g., allopregnanolone) for the subject. Prescribing can include drafting prescription forms, annotating medical records, etc.

[0014] The term "effective amount" or "pharmaceutically effective amount" refers to the amount and / or dosage and / or dosing regimen of one or more steroids or neurosteroids (e.g., allopregnanolone) necessary to produce a desired result, e.g., an amount sufficient to prevent, interrupt, or stop a seizure.

[0015] As used herein, the terms "treat" and "treatment" refer to delaying the onset of, slowing or reversing the progression of, reducing the severity of, or alleviating or preventing the disease or condition to which the term applies, or one or more symptoms of such disease or condition.

[0016] The terms "reduce," "suppress," "ameliorate," and "alleviate" refer to a detectable decrease in the frequency, severity, and / or duration of attacks. Reduction in attack frequency, severity, and / or duration can be measured by self-assessment (e.g., by patient report) or by a trained clinical observer. A determination of reduction in attack frequency, severity, and / or duration can be made by comparing the patient's condition before and after treatment.

[0017] The term "alleviate" refers to the reduction or elimination of one or more symptoms of the condition or disease, and / or the reduction in the incidence or delay in the onset or reduction in the severity of one or more symptoms (e.g., seizures) of the condition or disease, and / or the prevention of the condition or disease.

[0018] As used herein, the phrase "consisting essentially of" refers to the class or species of active pharmaceutical agents (e.g., neurosteroids, e.g., allopregnanolone) and excipients (e.g., hydroxypropyl-β-cyclodextrin or Captisol (sulfobutylether-β-cyclodextrin sodium salt)) included in a method or composition. In various embodiments, other unmentioned or unlisted active ingredients and inerts are specifically excluded. In various embodiments, additives (e.g., surfactants, acids (organic or fatty acids), alcohols, esters, cosolvents, solubilizers, lipids, polymers, glycols) are also specifically excluded.

[0019] The terms "subject," "individual," and "patient" refer interchangeably to mammals, preferably humans or non-human primates, but also to domesticated mammals (e.g., dogs or cats), laboratory animals (e.g., mice, rats, rabbits, hamsters, guinea pigs), and agricultural animals (e.g., horses, cows, pigs, sheep). In various embodiments, a subject can be a human (e.g., an adult male, adult female, adolescent male, adolescent female, boy, girl) receiving the care of a physician or other health care professional in a hospital, psychiatric facility, as an outpatient, or in other clinical settings. In certain embodiments, a subject may not be receiving the care or prescription of a physician or other health care professional.

[0020] The term "neuroactive steroid" or "neurosteroid" refers to steroid compounds that rapidly alter neuronal excitability by interacting with neurotransmitter-gated ion channels. Neurosteroids are compounds that rapidly alter neuronal excitability by interacting with GABA ANeurosteroids act as allosteric modulators of neurotransmitter receptors such as NMDA and sigma receptors. Neurosteroids can be used as sedatives for general anesthesia for surgical procedures, and can be used to treat epilepsy and traumatic brain injury. Exemplary neurosteroids include, for example, allopregnanolone, ganaxolone, alphaxolone, alphadolone, hydroxydione, minaxolone, and althesin (a mixture of alphaxolone and alphadolone). [The present invention 1001] 1. A method of treating, reducing, and / or alleviating symptoms associated with and / or caused by traumatic brain injury, Alzheimer's disease, epilepsy, anxiety, fragile X syndrome, post-traumatic stress disorder, lysosomal storage disorders (Niemann-Pick type C), depression (including postpartum depression), premenstrual dysphoric disorder, alcohol craving, and smoking cessation in a subject in need thereof, said method comprising administering to said subject a steroid. [The present invention 1002] 1. A method of treating, reducing and / or alleviating symptoms associated with and / or caused by epilepsy in a subject in need thereof, said method comprising administering to said subject a steroid. [The present invention 1003] 1. A method of promoting the cessation or interruption of an impending seizure in a subject in need thereof, comprising administering to said subject a steroid. [The present invention 1004] The method of any one of claims 1001 to 1003, wherein the steroid is a neurosteroid. [The present invention 1005] Any of the methods of claims 1001 to 1004, wherein the neurosteroid is selected from the group consisting of allopregnanolone, allotetrahydrodeoxycorticosterone, ganaxolone, alphaxolone, alphadolone, hydroxydione, minaxolone, and althesin. [The present invention 1006] 1005. The method of claim 1005, wherein the neurosteroid is allopregnanolone. [The present invention 1007] 1007. The method of any one of claims 1005 to 1006, wherein the steroid is formulated in a cyclodextrin. [The present invention 1008] The method of any of claims 1001 to 1007, wherein the steroid is formulated in hydroxypropyl-β-cyclodextrin, sulfobutylether-β-cyclodextrin sodium salt, or a mixture thereof. [The present invention 1009] Any of the methods of 1001 to 1008, wherein the subject is experiencing an aura. [The present invention 1010] Any of the methods of 1001 to 1009, wherein the subject has received a warning of an impending seizure. [The present invention 1011] The method of any of claims 1001 to 1010, wherein the subject is experiencing a seizure. [The present invention 1012] The method of any of claims 1001 to 1011, wherein the subject has status epilepticus. [The present invention 1013] 1012. The method of any one of claims 1001 to 1011, wherein the subject has myoclonic epilepsy. [The present invention 1014] The method of any of claims 1001 to 1011, wherein the subject is suffering from a cluster of seizures. [The present invention 1015] The method of any one of claims 1001 to 1014, wherein the seizure is a tonic seizure. [The present invention 1016] Any of the methods of 1001 to 1014, wherein the seizure is a clonic seizure. [The present invention 1017] The method of any one of claims 1001 to 1016, wherein the subject is a human. [The present invention 1018] The method of any of claims 1001 to 1017, wherein the steroid is administered intramuscularly (im), subcutaneously (sc) or intravenously (iv). [The present invention 1019] The method of any of claims 1001 to 1018, wherein the steroid is administered at a dose in the range of about 0.25 mg / kg to about 15 mg / kg. [The present invention 1020] Any of the methods of claims 1001 to 1019, comprising the step of treating, reducing, and / or alleviating symptoms associated with and / or caused by epilepsy by administering allopregnanolone formulated in sulfobutylether-β-cyclodextrin sodium salt intramuscularly (im), subcutaneously (sc), or intravenously (iv). [The present invention 1021] The method of any one of claims 1001 to 1020, wherein the epilepsy is status epilepticus. [The present invention 1022] A composition comprising or consisting essentially of a steroid and a cyclodextrin. [The present invention 1023] The composition of claim 1022, wherein the steroid is a neurosteroid. [The present invention 1024] The composition of claim 1023, wherein the neurosteroid is selected from the group consisting of allopregnanolone, allotetrahydrodeoxycorticosterone, ganaxolone, alphaxolone, alphadolone, hydroxydione, minaxolone, and althesin. [The present invention 1025] The composition of any one of claims 1022 to 1024, wherein the steroid is allopregnanolone. [The present invention 1026] The composition of any one of claims 1022 to 1025, wherein the cyclodextrin is hydroxypropyl-β-cyclodextrin, sulfobutylether-β-cyclodextrin sodium salt, or a mixture thereof. [The present invention 1027] 1027. The composition of any of claims 1022 to 1026, which is formulated for intramuscular (im), subcutaneous (sc) or intravenous (iv) administration. [The present invention 1028] Any of the compositions of 1022 to 1027 of the present inventions, comprising allopregnanolone and sulfobutylether-β-cyclodextrin sodium salt. [Brief explanation of the drawings]

[0021] [Figure 1] 1 illustrates the time course of protection by allopregnanolone (5α,3α-P) administered intravenously at doses of 1.5 mg / kg and 0.5 mg / kg, respectively, in a 6 Hz electrical stimulation (32 mA, 3 seconds) model. The interval between steroid injection and electrical stimulation is plotted on the abscissa, and the percentage of animals protected from seizures is plotted on the ordinate. Each point represents 8 mice. [Figure 2] Figure 1 illustrates the time course of protection by allopregnanolone (5α,3α-P) administered im at doses of 6, 3, and 1.5 mg / kg in a 6 Hz electrical stimulation (32 mA, 3 seconds) model. The interval between steroid injection and electrical stimulation is plotted on the abscissa, and the percentage of animals protected from seizures is plotted on the ordinate. Each point represents at least 8 mice. [Figure 3] 1 illustrates the time course of protection by allopregnanolone (5α,3α-P) administered sc at doses of 6 mg / kg and 1.5 mg / kg in a 6 Hz electrical stimulation (32 mA, 3 seconds) model. The interval between steroid injection and electrical stimulation is plotted on the abscissa, and the percentage of animals protected from seizures is plotted on the ordinate. Each point represents 8 mice. [Figure 4] Figure 1 illustrates the time course of protection by allopregnanolone (5α,3α-P) administered p.o. at doses of 300 mg / kg and 200 mg / kg (double the doses of 150 mg / kg and 100 mg / kg suspended / diluted in canola oil) in a 6 Hz electrical stimulation (32 mA, 3 seconds) model. The interval between steroid injection and electrical stimulation is plotted on the abscissa, and the percentage of animals protected from seizures is plotted on the ordinate. Each point represents 7-8 mice. [Figure 5]Figure 1 illustrates the effect of intravenous administration of allopregnanolone (5α,3α-P) (0.1–1.5 mg / kg) on ​​the occurrence of myoclonic twitches, generalized clonus, and tonic extension in response to PTZ (80 mg / kg, i.p.) injection in mice. 5α,3α-P was administered intravenously 1 min before PTZ injection. Bars represent the mean SEM of values ​​from eight mice. p<0.05 compared with vehicle control (ANOVA followed by Dunnett's test). [Figure 6] Figure 1 illustrates the effect of intravenous administration of allopregnanolone (5α,3α-P) (0.1–1.5 mg / kg) on ​​the occurrence of myoclonic twitches, generalized clonus, and tonic extension in response to PTZ (80 mg / kg, i.p.) injection in mice. 5α,3α-P was administered intravenously 2 min before PTZ injection. Bars represent the mean SEM of values ​​from eight mice. p<0.05 compared with vehicle control (ANOVA followed by Dunnett's test). [Figure 7] Figure 1 illustrates the effect of intravenous administration of allopregnanolone (5α,3α-P) (0.25–1.5 mg / kg) on ​​the occurrence of myoclonic twitches, generalized clonus, and tonic extension in response to PTZ (80 mg / kg, i.p.) injection in mice. 5α,3α-P was administered intravenously 30 min before PTZ injection. Bars represent the mean SEM of values ​​from eight mice. p<0.05 compared with vehicle control (ANOVA followed by Dunnett's test). [Figure 8] Figure 1 illustrates the effect of im administration of allopregnanolone (5α,3α-P) (0.25–1.5 mg / kg) on ​​the occurrence of myoclonic twitches, generalized clonus, and tonic extension in response to PTZ (80 mg / kg, i.p.) injection in mice. 5α,3α-P was administered im 2 min before PTZ injection. Bars represent the mean SEM of values ​​from at least seven mice. p<0.05 compared with vehicle control (ANOVA followed by Dunnett's test). [Figure 9]Figure 1 illustrates the effect of im administration of allopregnanolone (5α,3α-P) (0.25–1.5 mg / kg) on ​​the occurrence of myoclonic twitches, generalized clonus, and tonic extension in response to PTZ (80 mg / kg, i.p.) injection in mice. 5α,3α-P was administered im 30 min before PTZ injection. Bars represent the mean SEM of values ​​from eight mice. p<0.05 compared with vehicle control (ANOVA followed by Dunnett's test). [Figure 10] Figure 1 illustrates the time-concentration profile for plasma allopregnanolone (5α,3α-P) after a single iv injection in rats. Rats with indwelling jugular catheters received a single iv injection of 5α,3α-P or vehicle, and serial blood samples were collected at 1, 2, 10, 15, 30, 60, and 120 minutes post-injection. Plasma was assayed for 5α,3α-P by LC-MS. Each point represents at least four animals. DETAILED DESCRIPTION OF THE INVENTION

[0022] Detailed Description 1. Introduction Treatment of status epilepticus requires rapid administration of anti-seizure medications, which are typically delivered by either the intravenous (IV) or intramuscular (IM) route. Allopregnanolone (3α-hydroxy-5α-pregnan-20-one; 5α,3α-P) inhibits GABAergic activity. A Allopregnanolone, a steroid derived from endogenous progesterone, is a positive allosteric modulator of the progesterone receptor and a potent antiseizure agent with potential in the treatment of status epilepticus. This study determined and demonstrates that allopregnanolone administration provides protection from seizures when delivered intravenously (iv), intramuscularly (im), subcutaneously (sc), or orally (po).

[0023] 2. What can benefit you? In various embodiments, the subject has a condition that can be treated or alleviated by administration of a neurosteroid, such as allopregnanolone. Allopregnanolone has numerous medical uses, such as treating, reducing, and / or alleviating symptoms associated with and / or caused by traumatic brain injury, Alzheimer's disease, epilepsy, anxiety, fragile X syndrome, post-traumatic stress disorder, lysosomal storage disorders (Niemann-Pick type C), depression (including postpartum depression), premenstrual dysphoric disorder, alcohol craving, and smoking cessation. The subject may or may not be symptomatic.

[0024] Accordingly, the present invention also contemplates methods of treating, reducing, and / or alleviating symptoms associated with and / or caused by traumatic brain injury, Alzheimer's disease, epilepsy, anxiety, fragile X syndrome, post-traumatic stress disorder, lysosomal storage disorders (Niemann-Pick type C), depression (including postpartum depression), premenstrual dysphoric disorder, alcohol craving, and smoking cessation by administering a steroid or neurosteroid (e.g., allopregnanolone) as described herein dissolved or suspended in a vehicle suitable for systemic administration (e.g., intramuscular, intravenous, subcutaneous).

[0025] In some embodiments, the subject has epilepsy, has a history of epileptic seizures, or is currently experiencing epileptic seizures. In various embodiments, the patient may experience electrographic or behavioral seizures, or may experience aura (which is itself a localized seizure that can spread to a generalized behavioral seizure). For example, the subject may experience aura warning of the imminent onset of a seizure or a cluster of seizures.

[0026] Alternatively, the subject may be using a seizure prediction device that warns of the impending occurrence of a seizure or seizure cluster. Implantable seizure prediction devices are known in the art and are described, for example, in D'Alessandro et al., IEEE TRANSACTIONS ON BIOMEDICAL ENGINEERING, VOL. 50, NO. 5, MAY 2003, and U.S. Patent Application Publication Nos. 2010 / 0198098, 2010 / 0168603, 2009 / 0062682, and 2008 / 0243022.

[0027] The subject may have a personal or family history of any of the epilepsy conditions described herein.The subject may have been diagnosed with any of the epilepsy conditions described herein.In some embodiments, the subject has or is at risk of suffering from myoclonic seizures or myoclonic epilepsy, such as juvenile myoclonic epilepsy.The PTZ seizure model demonstrated herein is predicted to be useful and / or active in combating myoclonic seizures or myoclonic epilepsy in humans.

[0028] In various embodiments, the subject may be at risk of or have been exposed to a nerve agent or pesticide that can cause seizures. Exemplary nerve agents that can cause seizures include, for example, organophosphate nerve agents, such as tabun, sarin, soman, GF, VR, and / or VX. Exemplary pesticides that can cause seizures include, for example, organophosphate pesticides (e.g., acephate (Orthene), azinphos-methyl (Gusathion, Guthion), bensulide (Betasan, Lescosan), bomil (Swat), bromophos (Nexion), bromophos-ethyl (Nexagan), cadusafos (Apache, Ebufos, Rugby), carbophenothion (Trithion), chlorethoxyphos (Fortress), chlorfenvinphos (Apachlor), and the like. , Birlane), chlormephos (Dotan), chlorphoxim (Baythion-C), chlorpyrifos (Brodan, Dursban, Lorsban), chlorthiophos (Celathion), coumaphos (Asuntol, Co-Ral), crotoxyphos (Ciodrin, Cypona), crufomate (Ruelene), cyanofenphos (Surecide), cyanophos (Cyanox), cythioate (Cyflee, Proban), DEF (De-Green), EZ-Off D), Demeton (Systox), Demeton-S-methyl (Duratox, Metasystoxl), Diarifol (Torak), Diazinon, Dichlorofenthion (VC-13 Nemacide), Dichlorvos (DDVP, Vapona), Dicrotophos (Bidrin), Dimefos (Hanane, Pestox)XIV), dimethoate (Cygon, DeFend), dioxathion (Delnav), disulfoton (Disyston), ditalimfos, edifenphos, endothion, EPBP (S-seven), EPN, ethion (Ethanox), ethoprop (Mocap), ethyl parathion (E605, parathion, thiophos), etrimphos (Ekamet), famfur (Bash, Bo-Ana, Famfos), fenamiphos (Nemacur), fenitrothion (Accothion, Agrothion, Sumithion), fenophosphon (Agritox, trichloronate), fensulfothion (Dasanit), Enthion (Baytex, Entex, Tiguvon), fonofos (Dyfonate, N-2790), formothion (Anthio), fostietan (Nem-A-Tak), heptenophos (Hostaquick), hyometon (Ekatin), phosalone (Zolone), IBP (Kitazin), iodofenphos (Nuvanol-N), isazophos (Brace, Miral, Triumph), isofenphos (Amaze, Oftanol), isoxathion (E-48, Karphos), leptophos (Phosvel), malathion (Cythion), mephosphorane (Cytrolane), merphos (Easy Off-D, Folex), methamidophos (Monitor), methidathion (Supracide, Ultracide), methyl parathion (E601, Penncap-M), methyltrithion, mevinphos (Duraphos, Phosdrin), mipafox (Isopestox, Pestox)XV), monocrotophos (Azodrin), naled (Dibrome), oxydemeton-methyl (Metasystox-R), oxydeprophos (Metasystox-S), fencapton (G28029), phenthoate (dimephenthoate, phenthoate), phorate (Rampart, Thimet), phosalone (Azofene, Zolone), phospholan (Cylan, Cyolane), phosmet (Imidan, Prolate), phosphamidon (Dimecron), fostebpirim (Aztec), phoxim (Baythion), pirimiphos-ethyl (Primicid), pirimiphos-methyl (Actellic), profenofos (Curacron), propetamphos (Safrotin), propylthio These include pyrophosphate (Aspon), protoate (Fac), pyrazophos (Afugan, Curamil), pyridaphenthion (Ofunack), quinalphos (Bayrusil), Ronnel (fenchlorphos, Korlan), Schladan (OMPA), sulfotep (Bladafum, Dithione, Thiotepp), sulprofos (Bolstar, Helothion), temephos (Abate, Abathion), terbufos (Contraven, Counter), tetrachlorvinphos (Gardona, Rabon), tetraethyl pyrophosphate (TEPP), triazophos (Hostathion), and trichlorfon (Dipterex, Dylox, Neguvon, Proxol).

[0029] 3. Steroids The compositions generally comprise or consist essentially of a steroid, e.g., a neurosteroid, suspended or dissolved in a vehicle suitable for systemic administration, such as a cyclodextrin, e.g., hydroxypropyl-β-cyclodextrin or sulfobutylether-β-cyclodextrin sodium salt, or a mixture thereof.

[0030] In various embodiments, the neurosteroid is allopregnanolone (ALP).Also known as 3α-hydroxy-5α-pregnan-20-one or 3α,5α-tetrahydroprogesterone, IUPAC name 1-(3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl)ethanone, and CAS number 516-54-1, it is the prototype neurosteroid that exists in blood and brain.It is a metabolite of progesterone and acts as a GABA receptor. A Allopregnanolone is a GABA receptor modulator. A All GABA receptors, as well as receptor-active neurosteroids, such as allotetrahydrodeoxycorticosterone (3α,21-dihydroxy-5α-pregnan-20-one; THDOC), A Allopregnanolone positively regulates GABA receptor isoforms, with the isoform containing the δ-subunit exhibiting greater potentiation. AIt has similar pharmacological properties to other positive modulators of the receptor, such as anxiolytic and anticonvulsant activity. Allopregnanolone has been shown to be effective in treating Alzheimer's disease (Wang et al., Proc Natl Acad Sci USA. 2010 Apr 6;107(14):6498-503), cerebral edema (Limmroth et al., Br J Pharmacol. 1996 Jan;117(1):99-104) and traumatic brain injury (He et al., Restor Neurol Neurosci. 2004;22(1):19-31, and He, et al., Exp Neurol. 2004 Oct;189(2):404-12), mood disorders (Robichaud and Debonnel, Int J Neuropsychopharmacol. 2006 Apr;9(2):191-200), and type C Niemann-Pick disease (Griffin et al., Nat Med. 2004 It has neuroprotective effects in animal models of many neurodegenerative conditions, including those of the pentylenetetrazole (PTZ) model (Kokate et al., J Pharmacol Exp Ther. 1994 Sep;270(3):1223-9), and acts as an anticonvulsant against chemically induced seizures, such as the pentylenetetrazole (PTZ) model (Kokate et al., J Pharmacol Exp Ther. 1994 Sep;270(3):1223-9). The chemical structure of allopregnanolone is illustrated in Formula I below. TIFF0007744166000001.tif65128

[0031] In various embodiments, the compositions include allopregnanolone sulfates, salts, hemisuccinates, nitrosylates, derivatives, and the like.

[0032] Other neurosteroids that can be formulated in vehicles suitable for systemic administration include allotetrahydrodeoxycorticosterone (3α,21-dihydroxy-5α-pregnan-20-one; THDOC), 3α,21-dihydroxy-5β-pregnan-20-one, pregnanolone (3α-hydroxy-5β-pregnan-20-one), ganaxolone (INN, also known as CCD-1042; IUPAC name (3α,5α)-3-hydroxy-5-methylpregnan-20-one; 1-[(3R,5S,8R,9S,10S,13 S,14S,17S)-3-hydroxy-3,10,13-trimethyl-1,2,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydrocyclopenta[a]phenanthren-17-yl]ethanone), alphaxolone, alphadolone, hydroxydione, minaxolone, and artesin (a mixture of alphaxolone and alphadolone), tetrahydrodeoxycorticosterone, pregnenolone, dehydroepiandrosterone (DHEA), 7-substituted benzo[e]indene-3-carbonitriles (e.g., Hu, et al., J Med Chem. (1993) 36(24):3956-67; 7-(2-hydroxyethyl)benz[e]indene analogs (see, e.g., Han, et al., J Med Chem. (1995) 38(22):4548-56); 3α-hydroxy-5α-pregnan-20-one and 3α-hydroxy-5β-pregnan-20-one analogs (see, e.g., Han, et al., J Med Chem. (1996) 39(21):4218-32); enantiomers of dehydroepiandrosterone sulfate, pregnenolone sulfate, and (3α,5β)-3-hydroxypregnan-20-one sulfate (see, e.g., Nilsson, et al., J Med Chem. (1998) 41(14):2604-13); 13,24-cyclo-18,21-dinorcholane analogs (see, e.g., Jiang, et al., J Med Chem. (2003) 46(25):5334-48); N-acylated 17a-aza-D-homosteroid analogs (see, e.g., Covey, et al., J Med Chem.(2000) 43(17):3201-4); 5β-methyl-3-ketosteroid analogs (see, e.g., Zeng, et al., J Org Chem. (2000) 65(7):2264-6); 18-norandrostan-17-one analogs (see, e.g., Jiang, et al., J Org Chem. (2000) 65(11):3555-7); (3α,5α)- and (3α,5β)-3-hydroxypregnan-20-one analogs (see, e.g., Zeng, et al., J Med Chem. (2005) 48(8):3051-9); benzo[f]indenes (see, e.g., Scaglione, et al., J Med Chem. (2006) 49(15):4595-605); androgen enantiomers (see, e.g., Katona, et al., Eur J Med Chem. (2008) 43(1):107-13); cyclopenta[b]phenanthrene and cyclopenta[b]anthracene (see, e.g., Scaglione, et al, J Med Chem. (2008) 51(5):1309-18); 2β-hydroxygonane derivatives (see, e.g., Wang, et al, Tetrahedron (2007) 63(33):7977-7984); Δ16-alphaxalone and the corresponding 17-carbonitrile analogues (see, e.g., Bandyopadhyaya, et al, Bioorg Med Chem Lett. (2010) 20(22):6680-4); Δ(16)-alphaxalone and the Δ(17(20)) analogues of Δ(16)-alphaxalone (see, e.g., Stastna, et al, J Med Chem.(2011) 54(11):3926-34); neurosteroid analogs developed by CoCensys (now Purdue Neuroscience) (e.g., CCD-3693, Co2-6749 (also known as GMA-839 and WAY-141839); neurosteroid analogs described in U.S. Pat. No. 7,781,421 and PCT Patent Publications WO2008 / 157460; WO1993 / 003732; WO1993 / 018053; WO1994 / 027608; WO1995 / 021617; WO1996 / 016076; WO1996 / 040043, and salts, hemisuccinates, nitrosylates, sulfates, and derivatives thereof, but are not limited to these.

[0033] In various embodiments, the steroid or neurosteroid is not a sex hormone. In various embodiments, the steroid or neurosteroid is not progesterone.

[0034] The steroid or neurosteroid (e.g., allopregnanolone) may or may not be micronized, as appropriate. The steroid or neurosteroid (e.g., allopregnanolone) may or may not be encapsulated in microspheres suspended in oil, as appropriate.

[0035] 4. Formulation and Administration In various embodiments, steroids and / or analogs thereof can be administered systemically, as appropriate or desired, for example, intramuscularly (IM) or depot IM, subcutaneously (SQ), and depot SQ. In various embodiments, the dosage form is selected to facilitate delivery to the brain (for example, crossing the blood-brain barrier). In this way, the steroids or neurosteroids (for example, allopregnanolone) described herein can be easily delivered to the brain. Dosage forms known to those skilled in the art are suitable for steroid delivery.

[0036] A composition containing a therapeutically effective amount of steroid or neurosteroid (e.g., allopregnanolone) is provided. The steroid or neurosteroid (e.g., allopregnanolone) is preferably formulated into a suitable pharmaceutical preparation, such as a tablet, capsule, or elixir for oral administration, or a sterile solution or suspension for parenteral administration. Typically, the steroid or neurosteroid (e.g., allopregnanolone) is formulated into a pharmaceutical composition using techniques and methods well known in the art.

[0037] These steroids or neurosteroids (e.g., allopregnanolone) or analogs thereof can be administered in their "native" form or, if desired, in the form of salts, esters, amides, prodrugs, derivatives, etc., provided that said salts, esters, amides, prodrugs, or derivatives are appropriate and pharmaceutically effective, e.g., effective in the methods of the present invention. Salts, esters, amides, prodrugs, and other derivatives of active agents can be prepared using standard techniques known to those skilled in the art of synthetic organic chemistry, as described, for example, in March (1992) Advanced Organic Chemistry; Reactions, Mechanisms, and Structure, 4th Ed., NY Wiley-Interscience.

[0038] Methods for formulating such derivatives are known to those skilled in the art. For example, disulfide salts of some delivery agents are described in PCT Publication WO2000 / 059863, which is incorporated herein by reference. Similarly, acid salts of therapeutic peptides, peptoids, or other mimetics can be prepared from their free bases using conventional methods, which typically involve reaction with an appropriate acid. Generally, the base form of the drug is dissolved in a polar organic solvent such as methanol or ethanol, and an acid is added thereto. The resulting salt precipitates or can be precipitated by adding a less polar solvent. Suitable acids for preparing acid addition salts include, but are not limited to, organic acids such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, orotic acid, and inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Acid addition salts can be reconverted to free bases by treatment with an appropriate base. Particularly preferred acid addition salts of the active agents herein include those that can be prepared using halide salts, such as hydrochloric acid or hydrobromic acid. Conversely, base salts of the active agents of the present invention can be similarly prepared using pharmaceutically acceptable bases such as sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide, trimethylamine, and the like. In certain embodiments, base salts include alkali metal salts, such as sodium and copper salts.

[0039] To prepare salt forms of basic drugs, the pKa of the counterion is preferably at least about 2 pH lower than the pKa of the drug. Similarly, to prepare salt forms of acidic drugs, the pKa of the counterion is preferably at least about 2 pH higher than the pKa of the drug. This allows the counterion to lower the solution pH below the pHmax and reach the salt plateau, where the solubility of the salt exceeds that of the free acid or free base. This generalized rule regarding the pKa unit difference between the ionizable groups in the active pharmaceutical ingredient (API) and the ionizable groups in the acid or base makes proton transfer energetically favorable. If the pKa difference between the API and the counterion is not large enough, even if a solid complex is formed in an aqueous environment, it will rapidly disproportionate (e.g., decompose into individual entities of the drug and counterion).

[0040] Preferably, the counterion is a pharmaceutically acceptable counterion. Suitable anionic salt forms include acetate, benzoate, benzilate, bitartrate, bromide, carbonate, chloride, citrate, edetate, edisylate, estolate, fumarate, gluceptate, gluconate, hydrobromide, hydrochloride, iodide, lactate, lactobionate, malate, maleate, mandelate, mesylate, methyl bromide, methyl sulfate, mucate, napsylate, nitrate, pamoate (embonate), phosphate, and diphosphate. Suitable salt forms include, but are not limited to, salicylate and disalicylate, stearate, succinate, sulfate, tartrate, tosylate, triethiodide, valerate, and the like, while suitable cationic salt forms include, but are not limited to, aluminum, benzathine, calcium, ethylenediamine, lysine, magnesium, meglumine, potassium, procaine, sodium, tromethamine, zinc, and the like.

[0041] In various embodiments, the preparation of esters typically involves functionalization of hydroxyl and / or carboxyl groups present in the molecular structure of the active agent. In certain embodiments, esters are typically acyl-substituted derivatives of free alcohol groups, such as moieties derived from carboxylic acids of the formula RCOOH, where R is alkyl, preferably lower alkyl. If desired, esters can be reconverted to free acids using conventional hydrogenolysis or hydrolysis methods.

[0042] Amides may also be prepared using techniques known to those skilled in the art or described in the pertinent literature, for example, they may be prepared from esters, using suitable amine reactants, or they may be prepared from anhydrides or acid chlorides by reaction with ammonia or a lower alkyl amine.

[0043] The determination of effective amount when administered in a single dose is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure provided herein.Generally, the effective amount, i.e., effective dose, of steroid or neurosteroid (e.g., allopregnanolone) is determined by first administering a low dose, i.e., a small amount of the active substance, and then gradually increasing the administered dose, i.e., dosage, by adding a second or third drug treatment as necessary, until the desired effect in the treated subject is observed with minimal or no toxic side effects.Applicable methods for determining the appropriate dose and administration schedule for administering the combination of the present invention can be found, for example, in Brunton, et al., Goodman and Gilman's The Pharmacological Basis of Therapeutics, 12th Edition, 2010, McGraw-Hill Professional; Physicians' Desk Reference (PDR), 66 th Edition, 2012;Loyd, et al., Remington: The Science and Practice of Pharmacy, 22 stEd., 2012, Pharmaceutical Press; Martindale: The Complete Drug Reference, Sweetman, 2005, London: Pharmaceutical Press; and Martindale, Martindale: The Extra Pharmacopoeia, 31st Edition., 1996, Amer Pharmaceutical Assn., each of which is incorporated herein by reference. In various embodiments, the composition is formulated for oral administration at a dose ranging from about 5 mg / kg to about 250 mg / kg of steroid or neurosteroid (e.g., allopregnanolone), e.g., about 5 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, 40 mg / kg, 45 mg / kg, 50 mg / kg, 75 mg / kg, 100 mg / kg, 125 mg / kg, 150 mg / kg, 200 mg / kg, or 250 mg / kg.

[0044] In accordance with generally accepted pharmaceutical practice, approximately 1 to 1000 mg of a steroid or neurosteroid (e.g., allopregnanolone), or a physiologically acceptable salt or ester, is mixed with a physiologically acceptable vehicle, carrier, excipient, binder, preservative, stabilizer, flavor, etc. The amount of active substance in these compositions or preparations is such that a suitable dosage within the indicated range is obtained. The compositions are preferably formulated in a unit dosage form, each dosage containing about 1-1000 mg, 2-800 mg, 5-500 mg, 10-400 mg, 50-200 mg, for example about 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg or 1000 mg of the active ingredient. In various embodiments, the steroid or neurosteroid (e.g., allopregnanolone) is administered systemically (e.g., intramuscularly, intravenously, subcutaneously) at a dose ranging from about 0.25 mg / kg to about 15 mg / kg, e.g., about 0.25 mg / kg to about 15 mg / kg, e.g., about 0.25, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 mg / kg. The term "unit dosage form" refers to physically discrete units suitable as unit dosages for human subjects and other mammals, each unit containing a predetermined amount of active agent calculated to produce a desired therapeutic effect, together with suitable pharmaceutical excipients.

[0045] In various embodiments, the steroid or neurosteroid (e.g., allopregnanolone) is formulated for pulmonary administration. In various embodiments, the steroid or neurosteroid (e.g., allopregnanolone) is formulated for delivery by inhaler.

[0046] In various embodiments, steroid or neurosteroid (such as allopregnanolone) is atomized.Methods and systems for pulmonary delivery of steroid or neurosteroid (such as allopregnanolone) are known and useful in the art.Exemplary systems for aerosol delivery of steroid or neurosteroid (such as allopregnanolone) by inhalation are described in, for example, U.S. Patent Nos. 5,497,763; 5,660,166; 7,060,255; and 7,540,286; and U.S. Patent Application Publication Nos. 2003 / 0032638; and 2006 / 0052428, each of which is incorporated herein by reference in its entirety for all purposes.Preferably, steroid or neurosteroid (such as allopregnanolone) is atomized without applying heat.

[0047] When administering nebulized and / or aerosolized steroids or neurosteroids (e.g., allopregnanolone), the size of the aerosol particles can be within a suitable range for pulmonary delivery, particularly delivery to distal alveoli. In various embodiments, the aerosol particles have a mass median aerodynamic diameter ("MMAD") of less than about 5 μm, less than 4 μm, or less than 3 μm, e.g., in the range of about 1 μm to about 3 μm, e.g., in the range of about 2 μm to about 3 μm, e.g., in the range of about 0.01 μm to about 0.10 μm. Aerosols characterized by an MMAD in the range of about 1 μm to about 3 μm can be deposited on the alveolar walls by gravitational settling and absorbed into the systemic circulation, while aerosols characterized by an MMAD in the range of about 0.01 μm to 0.10 μm can also be deposited on the alveolar walls by diffusion. Aerosols characterized by an MMAD in the range of about 0.15 μm to about 1 μm are generally exhaled. Thus, in various embodiments, the aerosolized particulates can have an MMAD in the range of 0.01 μm to about 5 μm, e.g., in the range of about 0.05 μm to about 3 μm, e.g., in the range of about 1 μm to about 3 μm, e.g., in the range of about 0.01 μm to about 0.1 μm. Nebulized and / or aerosolized steroids or neurosteroids (e.g., allopregnanolone) can be delivered to the distal alveoli, allowing for rapid absorption and efficacy.

[0048] In various embodiments, the steroid or neurosteroid (e.g., allopregnanolone) is formulated in a solution containing an excipient suitable for aerosolized pulmonary delivery. The solution can include one or more pharmaceutically acceptable carriers and / or excipients. Pharmaceutically acceptable refers to being approved or capable of being approved by federal or state regulatory agencies for use in animals, more specifically humans, or listed in the United States Pharmacopoeia or other widely recognized pharmacopeia. The solution is preferably buffered to provide a relatively neutral pH, e.g., a pH in the range of about 4-8, e.g., a pH in the range of about 5-7. In some embodiments, the steroid or neurosteroid (e.g., allopregnanolone) is formulated in a buffered solution, e.g., phosphate-buffered saline.

[0049] In various embodiments, steroids or neurosteroids (e.g., allopregnanolone) are prepared as concentrated aqueous solutions. Conventional metered-dose liquid inhalers have low deep lung delivery efficiency because the particle size is not small enough (Kim et al., 1985 Am Rev Resp Dis 132:137-142; and Farr et al., 1995 Thorax 50:639-644). Therefore, these systems are mostly used for local drug delivery to the pulmonary airways. In addition, metered-dose inhalers may not be able to deliver a sufficient volume of even a concentrated steroid or neurosteroid (e.g., allopregnanolone) solution to produce the desired rapid anti-seizure effect. Therefore, in various embodiments, metered-dose inhalers are not used to deliver steroids or neurosteroids (e.g., allopregnanolone). In one embodiment, a nebulization system capable of delivering particles <5 μm (e.g., the PARI LC Star, which has a high efficiency, 78% respirable fraction 0.1-5 μm; see, e.g., pari.com) is used for pulmonary administration. Electronic nebulizers, which use a vibrating mesh or perforated plate to generate aerosols with the required particle size, can rapidly deliver sufficient amounts and may be useful (see, e.g., Knoch and Keller, 2005 Expert Opin Drug Deliv 2:377-390). Custom-made portable electronic nebulizers can also be made and may be useful.

[0050] Aerosolized delivery of steroids or neurosteroids (e.g., allopregnanolone) allows for the desired efficacy to be achieved with reduced dosage compared to, for example, intravenous or intranasal delivery. The appropriate dosage depends on the size and health of the patient and can be easily determined by a skilled clinician. The initial dose can be low and then gradually increased until the desired therapeutic effect is achieved with little or no adverse side effects. In various embodiments, the steroid or neurosteroid (e.g., allopregnanolone) is administered by the intrapulmonary route at a dose that is approximately 10%, 15%, 25%, 50%, or 75% of the dose established for their administration by other routes (e.g., oral, intravenous, or intranasal). In some embodiments, the steroid or neurosteroid (e.g., allopregnanolone) is administered by the intrapulmonary route at a dose in the range of about 0.05 mg / kg to about 1.0 mg / kg, e.g., about 0.2 mg / kg to about 0.8 mg / kg, e.g., about 0.05 mg / kg, 0.08 mg / kg, 0.1 mg / kg, 0.2 mg / kg, 0.3 mg / kg, 0.4 mg / kg, 0.5 mg / kg, 0.6 mg / kg, 0.7 mg / kg, 0.8 mg / kg, 0.9 mg / kg, or 1.0 mg / kg. In some embodiments, the steroid or neurosteroid (e.g., allopregnanolone) is administered by the intrapulmonary route at a dose ranging from about 10 μg / kg to about 80 μg / kg, e.g., about 20 μg / kg to about 60 μg / kg, e.g., about 25 μg / kg to about 50 μg / kg, e.g., about 10 μg / kg, 15 μg / kg, 20 μg / kg, 25 μg / kg, 30 μg / kg, 35 μg / kg, 40 μg / kg, 45 μg / kg, 50 μg / kg, 60 μg / kg, 70 μg / kg, or 80 μg / kg. In some embodiments, the steroid or neurosteroid (e.g., allopregnanolone) is administered by the intrapulmonary route at a dose ranging from about 0.3 μg / kg to about 3.0 μg / kg.

[0051] To prepare the composition, a steroid or neurosteroid (e.g., allopregnanolone) is mixed with a suitable pharmaceutically acceptable carrier. When the compound is mixed or added, the resulting mixture can be a solution, suspension, emulsion, or the like. Liposomal suspensions can also be suitable as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art. The form of the resulting mixture depends on several factors, such as the intended mode of administration and the solubility of the steroid or neurosteroid (e.g., allopregnanolone) in the selected carrier or vehicle. The effective concentration is sufficient to attenuate or ameliorate at least one symptom of the disease, disorder, or condition being treated and can be determined empirically.

[0052] Pharmaceutical carriers or vehicles suitable for administering the steroids or neurosteroids (e.g., allopregnanolone) provided herein include any carrier known to those skilled in the art to be suitable for the particular mode of administration (e.g., cyclodextrins). In addition, the active substance can be mixed with other active substances that do not impair the desired action, or with substances that complement the desired action or have a different action. The steroid or neurosteroid (e.g., allopregnanolone) can be formulated as the only pharmaceutically active ingredient in the composition, or can be combined with other active ingredients.

[0053] When steroids or neurosteroids (e.g., allopregnanolone) exhibit insufficient solubility, solubilization methods can be used. Such methods are known and include, but are not limited to, the use of cosolvents such as dimethyl sulfoxide (DMSO), the use of surfactants such as Tween™, and dissolution in aqueous sodium bicarbonate. Derivatives of steroids or neurosteroids (e.g., allopregnanolone), such as salts or prodrugs, can also be used to formulate effective pharmaceutical compositions.

[0054] The concentration of the steroid or neurosteroid (e.g., allopregnanolone) is effective to deliver, upon administration, an amount that is prophylactically effective and / or attenuates or ameliorates at least one symptom of the disorder for which the compound is administered. Typically, the composition is formulated for administration in a single dosage (e.g., daily).

[0055] Steroids or neurosteroids (e.g., allopregnanolone) can be prepared with carriers that protect them from rapid elimination from the body, such as sustained-release formulations or coatings. Such carriers include controlled-release formulations, such as, but not limited to, microencapsulated delivery systems. The active steroid or neurosteroid (e.g., allopregnanolone) is contained in a pharmaceutically acceptable carrier in an amount sufficient to exert a therapeutically useful effect in the treated patient without undesirable side effects. The therapeutically effective concentration can be determined experimentally by testing the steroid or neurosteroid (e.g., allopregnanolone) in known in vitro and in vivo model systems for the disorder being treated. Therapeutically or prophylactically effective doses can be determined by initially administering a low dose and then gradually increasing it until a dose is reached at which the desired effect is achieved with minimal or no undesirable side effects.

[0056] In various embodiments, the steroid or neurosteroid (e.g., allopregnanolone) and / or analog thereof can be packaged in a multi-dose or single-dose container. The packaged compounds and compositions can be provided, for example, in a kit containing components that can be assembled and used. For example, a lyophilized form of the compound inhibitor and a suitable diluent can be provided as separate components that are mixed before use. The kit can also include the compound inhibitor and a second therapeutic agent for co-administration. The inhibitor and the second therapeutic agent can be provided as separate components. The kit can include multiple containers, each containing a unit dose or multiple unit doses of the compound. The containers are preferably compatible with the desired mode of administration, such as, but not limited to, a parenteral depot product, a pre-filled syringe, an ampoule, or a vial.

[0057] The concentration and / or amount of steroid or neurosteroid (e.g., allopregnanolone) in the drug composition will depend on the absorption, inactivation, and excretion rates of the steroid or neurosteroid (e.g., allopregnanolone), the dosage regimen, and the amount administered, as well as other factors known to those skilled in the art.

[0058] The active ingredient may be administered at once, or may be divided into several small doses and administered at intervals.It is understood that the exact dosage and duration of treatment are a function of the disease being treated and can be determined experimentally using known testing protocols or by extrapolation from in vivo or in vitro test data.It should be noted that concentration and dosage values ​​may also vary depending on the severity of the condition to be alleviated.It should also be understood that for any particular embodiment, specific dosage regimens should be adjusted over time according to individual needs and the judgment of the professional administering or supervising the administration of the composition, and the concentration ranges described herein are merely exemplary and are not intended to limit the scope and practice of the compositions described in the claims.

[0059] 5. Efficacy Monitoring In various embodiments, administration of a steroid or neurosteroid (e.g., allopregnanolone) to a subject results in the prevention or alleviation of one or more symptoms of the disease state being treated (e.g., traumatic brain injury, Alzheimer's disease, epilepsy, anxiety, fragile X syndrome, post-traumatic stress disorder, lysosomal storage disorders (Niemann-Pick type C), depression (including postpartum depression), premenstrual dysphoric disorder, alcohol craving, and smoking cessation). Disease symptoms can be compared before and after administration of a steroid or neurosteroid (e.g., allopregnanolone) to a subject. Administration of a steroid or neurosteroid (e.g., allopregnanolone) to a subject is considered efficacious if the symptom no longer occurs (e.g., seizures) or is reduced, alleviated, and / or relieved after administration.

[0060] In various embodiments, administration of a steroid or neurosteroid (eg, allopregnanolone) to a subject results in the prevention of an impending attack and / or the termination or interruption of an ongoing attack.

[0061] In various embodiments, efficacy can be monitored by the subject.For example, when the subject experiences aura or receives a warning from the seizure prediction device, the subject can self-administer a single dose of steroid or neurosteroid (for example, allopregnanolone).When an effective amount of steroid or neurosteroid (for example, allopregnanolone) is administered, the aura sensation will subside and / or the seizure prediction device will no longer predict the imminent occurrence of an impending seizure.If the aura sensation does not subside and / or the seizure prediction device continues to predict an impending seizure, a second dose of steroid or neurosteroid (for example, allopregnanolone) can be administered.

[0062] In another embodiment, efficacy is monitored by caregivers.For example, when a subject is experiencing a seizure or a seizure has already begun, the subject may need to be administered steroid or neurosteroid (for example, allopregnanolone) by caregivers.If steroid or neurosteroid (for example, allopregnanolone) is administered in an effective amount, the seizure should stop or cease, along with the subject's seizure symptoms.If the seizure does not stop, a second dose of steroid or neurosteroid (for example, allopregnanolone) can be administered. [Example]

[0063] The following examples are offered to illustrate the claimed invention and are not intended to limit its scope.

[0064] Example 1: Anticonvulsant activity of intravenous and intramuscular allopregnanolone principle: Treatment of status epilepticus requires rapid administration of antiseizure medications, which are typically delivered by either the intravenous (iv) or intramuscular (im) route. Allopregnanolone (3α-hydroxy-5α-pregnan-20-one; 5α,3α-P) inhibits GABAergic activity. A It is a steroid derived from endogenous progesterone, a positive allosteric modulator of the receptor, and a potent antiseizure agent with potential in the treatment of status epilepticus. The purpose of this study was to determine whether allopregnanolone, when delivered intravenously and im, provides protection from seizures.

[0065] method: A mouse 6-Hz seizure model and a mouse pentylenetetrazol seizure model were used. A solution of 5α,3α-P was prepared in 6% (0.5 and 1.5 mg / ml) sulfobutylether-β-cyclodextrin sodium salt (Captisol®) in 0.9% saline. The solution was injected intravenously or imally (1, 2, and 30 minutes, or 2 and 30 minutes, respectively) prior to 6-Hz electrical stimulation or administration of PTZ (80 mg / kg, i.p.). For the PTZ model, animals were observed for 30 minutes, and myoclonic twitches and clonic and tonic seizures were recorded. Anticonvulsant activity was assessed by the delay in the onset of seizure signs. Allopregnanolone plasma levels in rats were determined by LC-MS.

[0066] result: 5α,3α-P exerted protective activity in the 6 Hz test 1–15 min after iv injection (1.5 mg / kg) but was inactive at 30 min. In contrast, im administration (3 mg / kg) produced a delayed protective effect (within 2 min) and lasted <2 h. At a dose of 0.1 mg / kg iv, 5α,3α-P failed to significantly delay seizure onset in the PTZ model at any pretreatment time (1, 2, or 30 min). However, at a dose of 0.5 mg / kg, administration 1 min before PTZ significantly delayed myoclonic twitches and clonic seizures and prevented tonic seizures and death, which were exclusively associated with tonic seizures, in 62.5% of animals. When injected 2 min before PTZ, 5α,3α-P (0.5 mg / kg) similarly increased the time to seizure onset and prevented tonic seizures in 25% of animals.

[0067] 5α,3α-P at a dose of 1.5 mg / kg completely prevented tonic seizures and death when injected i.v. 1 and 2 min before PTZ. 0.25, 0.5, and 1.5 mg / kg of 5α,3α-P protected 0%, 50%, and 100% of animals, respectively, from tonic seizures when injected im. 5α,3α-P at a dose of 1.5 mg / kg im conferred significant protection from tonic seizures when injected 30 min before PTZ, but the same dose was inactive when injected i.v. 30 min before PTZ. In rats, iv bolus doses of 0.5 mg / kg and 1.0 mg / kg of 5α,3α-P produced mean peak plasma levels (2 min) of 337 ng / ml and 746 ng / ml, respectively, with pooled mean two component halftimes of 2 min and 22 min at both doses.

[0068] Conclusion: Our results demonstrate that iv 5α,3α-P provides very rapid but transient anticonvulsant activity. When injected im, 5α,3α-P acts relatively rapidly and has a longer duration of action. Parenteral 5α,3α-P may be useful for the acute treatment of seizures.

[0069] Detailed Method Animals. Male NIH Swiss mice (22–30 g) were used. All procedures used in these studies were conducted in accordance with the University of California, Davis, Institutional Animal Care and Use Committee animal experimentation guidelines, in strict adherence to the National Research Council's Guide for the Care and Use of Laboratory Animals (National Academy Press, Washington, DC; available online at nap.edu / readingroom / books / labrats / ).

[0070] Test Substances and Drug Administration. Allopregnanolone (3α-hydroxy-5α-pregnan-20-one; 5α,3α-P) was synthesized by SAFC Pharma Inc. (Madison, WI, USA), and Captisol (sulfobutylether-β-cyclodextrin sodium salt) was provided by Ligand Pharmaceuticals, Inc. (La Jolla, CA, USA). Solutions of 5α,3α-P were made in 6% (0.5 and 1.5 mg / ml) or 24% (6 mg / kg) sulfobutylether-β-cyclodextrin sodium salt (Captisol®) in 0.9% saline. The volume used for all injections was 10–20 ml / kg body weight. To establish the time course of 5α,3α-P protection in the 6 Hz electrical stimulation (32 mA, 3 s) model, 5α,3α-P (0.5–6 mg / kg) was administered intravenously (iv), intramuscularly (im), subcutaneously (sc), or orally (po) before electrical stimulation. For the PTZ seizure test, 5α,3α-P or vehicle was administered iv or im 1, 2, or 30 min before PTZ.

[0071] Seizure model 6-Hz seizure test (Kaminski, et al., Epilepsia (2004) 45:1-4): 3-second corneal stimuli (200-microsecond duration, 6-Hz, 32-mA monopolar rectangular pulses) were delivered by a constant-current device (ECT unit 5780; Ugo Basile, Comerio, Italy). After stimulation, animals assumed a "stunned" posture accompanied by rearing and spontaneous movements, which lasted for 60–120 seconds in untreated animals. The experimental endpoint was protection from seizures; animals were considered protected if they resumed their normal exploratory behavior within 10 seconds of stimulation.

[0072] Pentylenetetrazol seizure test (Kokate, et al., J Pharmacol Exp Ther (1994) 270:1223-9): Mice were intraperitoneally injected with PTZ (80 mg / kg) and observed for 30 minutes. The time to the onset of myoclonic twitches, clonus, and tonic extension was recorded.

[0073] Surgery and Blood Collection: Male rats were implanted with indwelling jugular catheters as described (Baumann, et al., J Neurosci. (1998) 18:9069-77). Animals were allowed to recover for at least 1 week. Experiments were performed while the animals were in their home cages. Rats received an intravenous injection of vehicle or 5α,3α-P, and serial blood samples were collected into chilled tubes at 1, 2, 10, 15, 30, 60, and 120 minutes after the intravenous injection. 5α,3α-P and D4-5α,3α-P (internal standard) were extracted from rat plasma by SPE. Extracted 5α,3α-P and D4-5α,3α-P were quantified by ultra-performance liquid chromatography (UPLC) / atmospheric pressure chemical ionization (APCI) / tandem mass spectrometry (MS / MS).

[0074] Data Analysis. Results are expressed as mean ± SEM. The significance of differences in responses of treatment groups compared to controls was based on one-way analysis of variance (ANOVA) followed by specific post-hoc comparisons using Dunnett's test. Differences were considered statistically significant when the probability of error was less than 0.05 (p<0.05).

[0075] The results are shown in Figures 1-10. Our results demonstrate that iv 5α,3α-P provides extremely rapid but transient anticonvulsant activity. When injected im, 5α,3α-P acts relatively rapidly and has a longer duration of action. The low bioavailability of 5α,3α-P after oral administration prolongs the time to peak effect and duration of action. Parenteral 5α,3α-P is useful for the acute treatment of seizures.

[0076] It is understood that the examples and embodiments described herein are for illustrative purposes only, and that various changes and modifications therein will be suggested to those skilled in the art, which are within the spirit and scope of the present application and the scope of the claims. All publications, patents, and patent applications mentioned herein are incorporated herein by reference in their entirety for all purposes.

Claims

1. A pharmaceutical composition comprising an effective amount of a composition comprising a neurosteroid or a physiologically acceptable salt thereof formulated with a physiologically acceptable carrier for treating, reducing, and / or alleviating postpartum depression in a subject in need thereof, wherein the neurosteroid consists of allopregnanolone and the carrier is a sulfobutylether-β-cyclodextrin salt, and the pharmaceutical composition is an intravenous injection of allopregnanolone formulated in the sulfobutylether-β-cyclodextrin salt.

2. The pharmaceutical composition of claim 1, wherein the subject is a human.

3. 2. The pharmaceutical composition of claim 1, wherein allopregnanolone is administered at a dose ranging from about 0.25 mg / kg to about 15 mg / kg.

4. 10. The pharmaceutical composition of claim 1, further comprising a buffering agent.

5. 5. The pharmaceutical composition of claim 4, which is buffered to a pH in the range of about 4 to 8, optionally in the range of about 5 to 7.

6. 6. The pharmaceutical composition of claim 4 or 5, wherein the buffer is phosphate buffered saline.

7. 10. The pharmaceutical composition of claim 1, formulated in a unit dosage form containing about 1 to 1000 mg of allopregnanolone.

8. 10. The pharmaceutical composition of claim 1, comprising 1.5 mg / ml allopregnanolone in 6% sulfobutylether-β-cyclodextrin.

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