US 2010.00566.17A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2010/0056617 A1 Steiner et al. (43) Pub. Date: Mar. 4, 2010
(54) ROLE OF LIMONOID COMPOUNDS AS Related U.S. Application Data NEUROPROTECTIVE AGENTS (62) Division of application No. 1 1/893,100, filed on Aug. 13, 2007. (75) Inventors: Joseph(US): Avindra P. Steiner, Nath, Baltimore, Ellicott City,MD (60) Pygal application No. 60/837.365, filed on Aug. MD (US); Norman Haughey, s Baltimore, MD (US) Publication Classification (51) Int. Cl. Correspondence Address: A63L/35 (2006.01) EDWARDS ANGELL PALMER & DODGE LLP A6IP 25/18 (2006.01) P.O. BOX SS874 A6IP 25/28 (2006.01) BOSTON, MA 02205 (US) (52) U.S. Cl...... S14/453 (57) ABSTRACT (73) Assignee: The Johns Hopkins University, Disclosed herein are neuroprotective compounds. Methods Baltimore, MD (US) for the preparation of Such compounds are disclosed. Also disclosed are pharmaceutical compositions that include the (21) Appl. No.: 12/590,270 compounds. Methods of using the compounds disclosed, alone or in combination with other therapeutic agents, for the (22) Filed: Nov. 5, 2009 treatment of neurodegenerative conditions are provided.
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ROLE OF LIMONOID COMPOUNDSAS 0005. In a further embodiment, the at least one compound NEUROPROTECTIVE AGENTS of Formula I has a structure selected from:
RELATED APPLICATIONS
0001. This application claims the benefit of U.S. Provi sional Patent Application No. 60/837.365, entitled “Role of Liminoid Compounds as Neuroprotective Agents.” filed Aug. 11, 2006, the contents of which are incorporated by reference in their entirety.
BACKGROUND OF THE INVENTION 0002 Neurodegenerative conditions such as Alzheimer's Disease, multiple sclerosis, AIDS-related dementia, Hunting ton's Disease, stroke, and spinal cord trauma are character ized by extensive loss of neurons or glia.
SUMMARY OF THE INVENTION 0003. Described herein are neuroprotective compounds. Also described herein are pharmaceutical formulations com prising Such neuroprotective compounds, and methods for 0006. In a further embodiment, the at least one compound using Such neuroprotective compounds in the prophylaxis of Formula I has the structure: and treatment of neurodegenerative conditions. 0004. Accordingly, in one aspect provided herein is a pharmaceutical composition comprising at least one com pound having the structure of Formula I:
wherein R and Rs together form a Substituted cycloalkyl or cycloalkenyl group. wherein each compound of Formula I is in a substantially 0007. In a further embodiment, the at least one compound purified form; and of Formula I has the structure: Y is O and Y, is OH, O-alkyl, O-(hydroxyalkyl) or O-(alkoxyalkyl); or Y and Y together form a furan group; RandR together form a substituted cycloalkyl or cycloalk enyl group; and / O is selected from s CH O , or s CH R6 \7 / O, O wherein X and R together form a substituted heteroalicyclic R X group provided that R is H; or pharmaceutically acceptable salts, esters, prodrugs, or metabolites thereof; wherein R and R, together form a substituted cycloalkyl or or ester derivatives, saccharide derivatives, or—(CH2CH2O) cycloalkenyl group; and CH derivatives thereof, where n is 1 to 100: X is selected from H, oxo, OH, O-alkyl, O-(hydroxyalkyl). and a pharmaceutically acceptable excipient O-(alkoxyalkyl), or O—C(O)-alkyl. US 2010/00566.17 A1 Mar. 4, 2010
0008. In a further embodiment, the at least one compound 0010. In a further embodiment, the at least one compound of Formula I has a structure selected from: of Formula I has the structure:
wherein R and R, together form a substituted cycloalkyl or cycloalkenyl group; and X is selected from H, oxo, OH, O-alkyl, O-(hydroxyalkyl). O-(alkoxyalkyl), or O—C(O)-alkyl. 0011. In a further embodiment, the at least one compound of Formula I has a structure selected from:
Ro Rs wherein Rs and R are independently H or alkyl: X and X are independently selected from H, oxo, OH, O-alkyl, O-(hydroxyalkyl), O-(alkoxyalkyl), or O C(O)- alkyl; and is selected from O V7 O 0009. In a further embodiment, the at least one compound of Formula I has the structure:
wherein R and Rs together form a Substituted cycloalkyl or cycloalkenyl group. US 2010/00566.17 A1 Mar. 4, 2010 wherein Rs and R are independently H or alkyl: 0015. In another aspect provided herein is a pharmaceuti X and X are independently selected from H, oxo, OH, cal composition comprising at least one compound having the O-alkyl, O-(hydroxyalkyl), O-(alkoxyalkyl), or O C(O)- structure of Formula II: alkyl; and is selected from O
V7 O
0012. In a further embodiment, the at least one compound of Formula I has the structure:
wherein Het is a Substituted or unsubstituted oxygen-con taining aromatic or non-aromatic heterocycle; L is a bond or an alkylene group; each ------is independently selected from s R10 O,
O s / \ s X and Rio togetherform a substituted heteroalicyclic group. V/ X24 R20 0013. In a further embodiment, the at least one compound of Formula I has the structure: provided that no two adjacent ------groups are adjacent
O
V7O
groups: each X2, X, X2, X, and X is independently selected from H, oxo, OH, OC(O)-alkyl, O-(hydroxyalkyl). O-(alkoxyalkyl), or O-alkyl: wherein RandR togetherform a Substituted cycloalkyl or cycloalkenyl group. each Ro R and R is selected from Horalkyl, or any two 0014. In a further embodiment, the at least one compound of X20, X2, X22, X2: X24, R20 R2, or R22 can form an of Formula I has the structure: optionally substituted oxygen-containing heterocycle; or pharmaceutically acceptable salts, esters, prodrugs, or metabolites thereof; or ester derivatives, saccharide derivatives, or—(CH2CH2O) CH derivatives thereof, where n is 1 to 100; and a pharmaceutically acceptable excipient. I0016. In a further embodiment, the Het is an unsubsti tuted furanyl group. In a further or alternative embodiment, L is a bond. I0017. In a further or alternative embodiment, RandR are CH. In a further embodiment at least one of groups is a . Inafurther or alternative embodi ment, at least one of ------groups is a R13 R14 wherein R and Ra are independently H or alkyl, V7 X is selected from H, oxo, OH, O-alkyl, O-(hydroxyalkyl). O O-(alkoxyalkyl), or O—C(O)-alkyl; and Rs is alkyl-C(O)O-alkyl. US 2010/00566.17 A1 Mar. 4, 2010
In a further or alternative embodiment, at least one of maceutically acceptable excipients. In a further or alternative groups is a embodiment, the pharmaceutical composition has atherapeu tically effective amount of a compound isolated from the plant families of order Rutales, including in Maliaceae and Rutaceae. In a further or alternative embodiment, the phar maceutical composition has a therapeutically effective amount of a compound derived from a 4.4.8-trimethyl-17 In a further or alternative embodiment, X is an oxo group. In furanylsteroid skeleton. In a further or alternative embodi a further or alternative embodiment, the compound of For ment, the pharmaceutical composition has a therapeutically mula II is selected from: effective amount of a compound is a tetranortriterpenoid. 0019. In another aspect provided herein is a method for treating or reducing the risk of a neurodegenerative condition in a Subject in need thereof by administering to the Subject a therapeutically effective amount of any of the above-de scribed neuroprotective compound compositions. In some embodiments, the sole active ingredient in the pharmaceuti cal composition administered to the Subject is a neuroprotec tive compound disclosed herein. In some embodiments, the composition to be administered to the Subject comprises a neuroprotective compound that at a concentration of 10 LM provides at least 6% (e.g., at least 20%, 50%, or 70%) pro tection against 3 mM 3-nitropropionic acid to rat mixed hip pocampal cultures. In some embodiments, the Subject is diag nosed as Suffering from the neurodegenerative condition prior to administration of the composition. In some embodi ments, administration of the composition to the Subject is parenteral, intravenous, Subcutaneous, intra-muscular, trans nasal, intra-arterial, transdermal, or respiratory. In some embodiments, the subject to be treated is administered, in addition to one of the above-described compositions, a com position comprising a therapeutically effective amount of a polyphenol (e.g., resveratrol or epigallocatechin 3-gallate) or an antioxidant compound (e.g., Vitamin C or Vitamin E). 0020. In a related aspect, the subject to be treated is suf fering from a chronic neurodegenerative condition. In some embodiments, the chronic neurodegenerative condition is Alzheimer's disease. In some embodiments, in addition to administering one of the above-described neuroprotective compositions to a Subject Suffering from Alzheimer's Dis ease, the level of one or more Alzheimer's Disease prognostic biomarkers is determined in a biological sample from the Subject. In some embodiments, one or more Alzheimer's Dis ease prognostic biomarkers to be assayed comprise tau pro tein, phospho-tau protein, B-amyloid peptide, B-amy loido peptide, C1q protein, IL-6 protein, ApoE protein, C-1-antichymotrypsin protein, oxysterol, isoprostane, 3-ni trotyrosine, or any combination thereof. In some embodi ments, the subject to be treated is suffering from multiple Sclerosis. In some embodiments, the Subject to be treated is Suffering from Huntington's disease. In some embodiments, the subject to be treated is suffering from AIDS-related dementia. In some embodiments, the subject to be treated is Suffering from Schizophrenia. In some embodiments, the Sub ject to be treated is suffering from Amyotrophic Lateral Scle rosis. In some embodiments, the subject to be treated is suf fering from a retinal disease. In some embodiments, the Subject to be treated is Suffering from glaucoma, optic neuri tis, compressive optic neuropathy, or a hereditary neuropathy. In some embodiments, the subject to be treated is suffering from epilepsy. 0021. In a related aspect, the subject to be treated is suf 0018. In a further or alternative embodiment, the pharma fering from an acute neurodegenerative condition. In some ceutical composition has a therapeutically effective amount embodiments, the subject to be treated is suffering from a of a compound presented in Tables 2 or 3, along with phar stroke (e.g., an acute thromboembolic stroke, a focal
US 2010/00566.17 A1 Mar. 4, 2010 disease, Schilder's disease, Spielmeyer-Vogt-Sjogren-Batten 0040. Where a neurodegenerative disorder affects a cog disease (also known as Batten disease), Spinocerebellar nitive ability, a Subject can be diagnosed by any one of a ataxia (multiple types with varying characteristics), Spinal number of standardized cognitive assays, e.g., the Mini-Men muscular atrophy, Steele-Richardson-Olszewsli disease, tal State Examination, the Blessed Information Memory Con Tabes dorsalis, or any combination thereof. centration assay, or the Functional Activity Questionnaire. 0037. In some embodiments, the methods described See, e.g., Adelman et al. (2005), Am. Family Physician, 71 (9): herein can be used to treat acute neurodegenerative condi 1745-1750. Indeed, in some cases a subject can also be diag tions, which include, but are not limited to stroke (e.g., throm nosed as having a high risk of developing a chronic neurode boembolic stroke, focal ischemia, global ischemia, or tran generative condition (e.g., Alzheimer's disease), even in the sient ischemic attack), ischemia resulting from a Surgical absence of overt symptoms. For example, the risk of Alzhe technique involving prolonged halt of blood flow to the brain, imer's disease in a Subject can be determined by detecting a head trauma, spinal trauma, or any combination thereof. decrease in the Volumes of the Subject's hippocampus and 0.038 Symptoms, diagnostic tests, and prognostic tests for amygdala, using magnetic resonance imaging. See, e.g., den each of the above-mentioned conditions are described in, e.g., Heijeretal. (2006), Arch Gen Psychiatry, 63(1):57-62. Assay the Diagnostic and Statistical Manual of Mental Disorders, of prognostic biomarkers in a sample from a Subject are also 4" ed., 1994, Am. Psych. Assoc.; and Harrison's Principles useful in prognosis or diagnosis of a chronic neurodegenera of Internal Medicine(C,' 16th ed., 2004, The McGraw-Hill tive condition. For example, where the chronic neurodegen Companies, Inc. erative condition is Alzheimer's disease, prognostic biomar 0039 For example, where the subject is at risk of or is kers include, but are not limited to, total tau protein, phospho Suffering from multiple Sclerosis, a set of Standard criteria, tau protein, f-amyloid peptide, f-amyloido peptide, such as the "McDonald Criteria' can be used for prognosis/ complement component 1. q Subcomponent (C1q) protein, diagnosis. See McDonald et al. (2001), Ann Neurol, 50(1): interleukin 6 (IL-6) protein, apolipoprotein E (APOE) pro 121-127. Magnetic resonance imaging (MM) of the brain and tein, C.-1-antichymotrypsin protein, oxysterol (e.g., 24S-hy spine can be used to evaluate individuals with Suspected droxycholesterol), isoprostane (e.g., an F2-isoprostane), multiple sclerosis. MRI shows areas of demyelination as 3-nitrotyrosine, homocysteine, or cholesterol, or any combi bright lesions on T2-weighted images or FLAIR (fluid attenu nation thereof, e.g., the ratio of B-amyloid peptide to ated inversion recovery) sequences. Gadolinium contrast is f-amyloido peptide. used to demonstrate active plaques on T1-weighted images. 0041. The type of biological sample utilized in prognostic Further, a prognostic biomarker assay of cerebrospinal fluid Alzheimer's biomarker assays will vary depending on the (CSF) obtained by lumbar puncture can provide evidence of prognostic biomarker to be measured. Further, the relation chronic inflammation of the central nervous system. Specifi ship between the level of a prognostic biomarker and Alzhe cally, CSF is tested for oligoclonal bands, which are immu imer's risk varies depending on the particular biomarker, as noglobulins found in 85% to 95% of people with definite MS, well as on the biological sample in which the level of the albeit not exclusively in MS patients. Additional criteria for biomarker is determined. In other words, the level of the diagnosis of multiple Sclerosis include, e.g., a reduction in biomarker in a biological sample may be directly correlated visual evoked potentials and Somatosensory evoked poten or inversely correlated with the risk of Alzheimer's Disease, tials, which are indicative of demyelination. as summarized in Table 1.
TABLE 1
ALZHEIMERS DISEASE PROGNOSTIC BIOMARKERS
Biological Correlation to Biomarker Sample Type Dementia Risk Reference tau protein cerebrospinal increased Hampel et al. (2004), Mol Psychiatry, 9: 705-710 fluid (CSF) phospho-tau protein CSF increased Hampel et al. (2004), Arch Gen Psychiatry, 61: 95-102 Hansson et al. (2006), Lancet Neural, 5(3): 228-234 3-amyloid peptide CSF decreased Hampel et al. (2004), Mol Psychiatry, 9: 705-710 Ratio off-amyloid plasma decreased Graff-Radford et al. (2007), Arch Neural, 64(3): 354-362; peptide to 3-amyloido CSF decreased Hansson et al. (2007), Dement Geriatr Cogn Disord, peptide 23(5):316-20 C1q protein CSF decreased Smyth et al. (1994), Neurobiol Aging, 15(5): 609-614 IL-6 protein plasma increased Licastro et al. (2000), J Neuroimmunol, 103: 97-102; CSF increased Sun et al. (2003), Dement Geriatr Cogn Disord, 6(3): 136-44 APOE protein CSF increased Fukuyama et al. (2000), Eur Neural, 43(3):161-169 C-1-antichymotrypsin plasma increased Diket al. (2005), Neurology, 64(8): 1371-1377. protein oxysterol CSF increased Papassotiropoulos et al. (2002), J Psychiatr Res, 36(1): 27-32 isoprostane CSF increased Montine et al. (2005), Antioxid Redox Signal, 7(1-2): 269-275 3-nitrotyrosine CSF increased Tohgi et al. (1999), Neurosci Lett, 269(1): 52-54 homocysteine plasma increased Seshadri et al. (2002), N Engl J Med, 346(7): 476–83 cholesterol plasma increased Panza et al. (2006), Neurobiol Aging, 27(7): 933-940 US 2010/00566.17 A1 Mar. 4, 2010
0042 Animal models are useful for establishing a range of and B (2001), Plenum Press, New York Unless otherwise therapeutically effective doses of neuroprotective com indicated, methods of mass spectroscopy, NMR, HPLC, pro pounds for treating any of the foregoing diseases. For tein chemistry, biochemistry, recombinant DNA techniques example, animal models of chronic neurodegenerative con and pharmacology are employed. Standard techniques can be ditions have been established, e.g., for Alzheimer's disease, used for chemical syntheses, chemical analyses, pharmaceu multiple Sclerosis, amyotrophic lateral Sclerosis, multiple tical preparation, formulation, and delivery, and treatment of system atrophy, and Huntington's disease. See, e.g., Spires et patients. Standard techniques can be used for recombinant al. (2005), NeuroRx., 203):447-464; Gold et al. (2006), Brain, DNA, oligonucleotide synthesis, and tissue culture and trans 129(8): 1953-1971; Wong et al. (2002), Nat. Neurosci., 5(7): formation (e.g., electroporation, lipofection). 633-639, Stefanova et al. (2005), Am J Pathol, 166(3): 869 0047. As used herein, C-C, includes C-C, C-C . . . 876, Tadros et al. (2005), Pharmacol Biochem Behav, 82(3): C-C. 574-582. These animal models develop a chronic 0048. An “alkyl group refers to a straight-chained, neurodegenerative condition that is manifested behaviorally branched or cyclic aliphatic hydrocarbon group. The “allyl by impaired learning, memory, or locomotion. Cognitive moiety may have 1 to 10 carbon atoms (whenever it appears abilities, as well as motor functions in non-human animals herein, a numerical range such as "1 to 10” refers to each Suffering from a chronic neurodegenerative condition can be integer in the given range; e.g., “1 to 10 carbonatoms” means assessed using a number of behavioral tasks. Well-established that the alkyl group may have 1 carbonatom, 2 carbonatoms, sensitive learning and memory assays include the Morris 3 carbon atoms, etc., up to and including 10 carbon atoms, Water Maze (MWM), context-dependent fear conditioning, although the present definition also covers the occurrence of cued-fear conditioning, and context-dependent discrimina the term “alkyl where no numerical range is designated). The tion. See, e.g., Anger (1991), Neurotoxicology, 12(3):403 alkyl group of the compounds described herein may be des 413. Locomotor behavior, e.g., following spinal trauma, is ignated as "C-C alkyl or similar designations. By way of commonly assessed using a 21-point open field locomotion example only, "C-C alkyl” indicates that there are one to score assay developed by Basso, Beattie, and Bresnahan four carbon atoms in the alkyl chain, i.e., the alkyl chain is (BBB) (Basso, et al. (1995), J. Neurotrauma, 12(1): 1-21). selected from among methyl, ethyl, propyl; iso-propyl. n-bu Such animal models are suitable for testing effective dose tyl, iso-butyl, sec-butyl, and t-butyl. Thus C-C alkyl ranges for the neuroprotective compounds and compositions includes C-C alkyl and C-C alkyl. Alkyl groups can be described herein as well as for identifying additional neuro Substituted or unsubstituted. Typical alkyl groups include, but protective compounds. are in no way limited to, methyl, ethyl, propyl, isopropyl. butyl, isobutyl, tertiary butyl, pentyl, hexyl, cyclopropyl. Certain Chemical Terminology cyclobutyl, cyclopentyl, cyclohexyl, and the like. 0043 All patents, patent applications, published materials 0049. An “alkoxy group refers to a (alkyl)O— group, referred to throughout the entire disclosure herein, unless where alkyl is as defined herein. noted otherwise, are incorporated by reference in their 0050 “Hydroxyalkyl refers to an alkyl radical, as defined entirety. In the event that there area plurality of definitions for herein, substituted with at least one hydroxy group. Non terms herein, those in this section prevail. Where reference is limiting examples of a hydroxyalkyl include, but are not made to a URL or other such identifier or address, it is under limited to, hydroxymethyl, 2-hydroxyethyl, 2-hydroxypro stood that such identifiers can change and particular informa pyl, 3-hydroxypropyl, 1-(hydroxymethyl)-2-methylpropyl. tion on the internet can come and go, but equivalent informa 2-hydroxybutyl, 3-hydroxybutyl, 4-hydroxybutyl, 2,3-dihy tion can be found by searching the internet. Reference thereto droxypropyl, 1-(hydroxymethyl)-2-hydroxyethyl, 2,3-dihy evidences the availability and public dissemination of such droxybutyl, 3,4-dihydroxybutyl and 2-(hydroxymethyl)-3- information. hydroxypropyl, 2-hydroxyethyl, 2,3-dihydroxypropyl, and 0044. It is to be understood that the foregoing general 1-(hydroxymethyl)-2-hydroxyethyl. description and the following detailed description are exem 0051 "Alkoxyalkyl refers to an alkyl radical, as defined plary and explanatory only and are not restrictive of any herein, Substituted with at least one alkoxy group, as defined Subject matter claimed. In this application, the use of the herein. singular includes the plural unless specifically stated other 0.052 An "amide' is a chemical moiety with the formula wise. It must be noted that, as used in the specification and the - C(O)NHR or -NHC(O)R, where R is selected from appended claims, the singular forms “a” “an and “the among alkyl, cycloalkyl, aryl, heteroaryl (bonded through a include plural referents unless the context clearly dictates ring carbon) and heteroalicyclic (bonded through a ring car otherwise. In this application, the use of “or” means “and/or bon). An amide moiety may form a linkage between an amino unless stated otherwise. Furthermore, use of the term “includ acid or a peptide molecule and a compound described herein, ing as well as otherforms, such as “include.” “includes, and thereby forming a prodrug. Anyamine, or carboxyl side chain “included, is not limiting. on the compounds described herein can be amidified. The 0045. The section headings used herein are for organiza procedures and specific groups to make Such amides can be tional purposes only and are not to be construed as limiting found in Greene and Wuts, Protective Groups in Organic the subject matter described. All documents, or portions of Synthesis, 3' Ed., John Wiley & Sons, New York, N.Y., 1999, documents, cited in the application including, but not limited which is incorporated herein by reference in its entirety. to, patents, patent applications, articles, books, manuals, and 0053. The term “bond' or “single bond” refers to a chemi treatises are hereby expressly incorporated by reference in cal bond between two atoms, or two moieties when the atoms their entirety for any purpose. joined by the bond are considered to be part of larger sub 0046) Definition of standard chemistry terms may be Structure. found in reference works, including Carey and Sundberg 0054) The term “carbocyclic” or “carbocycle” refers to a “ADVANCED ORGANIC CHEMISTRY 4' E.D.” Vols. A (2000) compound which contains one or more covalently closed ring US 2010/00566.17 A1 Mar. 4, 2010
structures, and that the atoms forming the backbone of the replaced with halogenatoms, the halogenatoms are not all the ring are all carbon atoms. The term thus distinguishes car same as one another. The terms “fluoroalkyl and “fluoro bocyclic from heterocyclic rings in which the ring backbone alkoxy include haloalkyl and haloalkoxy groups, respec contains at least one atom which is different from carbon. tively, in which the halo is fluorine. In certain embodiments, Carbocycles include cycloalkyls and aryls. haloalkyls are optionally substituted. 0055. The term “cycloalkyl” refers to a monocyclic or 0060. As used herein, the terms "heteroalkyl "heteroalk polycyclic radical that contains only carbon and hydrogen, enyl and "heteroalkynyl' include optionally substituted and may be saturated, partially unsaturated, or fully unsatur alkyl, alkenyl and alkynyl radicals in which one or more ated. Cycloalkyl groups include groups having from 3 to 10 skeletal chain atoms are selected from an atom other than ring atoms. Illustrative examples of cycloalkyl groups include carbon, e.g., oxygen, nitrogen, Sulfur, silicon, phosphorus or the following moieties: combinations thereof. 0061. The term "heteroatom” refers to an atom other than carbon or hydrogen. Heteroatoms are typically independently selected from among oxygen, Sulfur, nitrogen, silicon and phosphorus, but are not limited to these atoms. In embodi ments in which two or more heteroatoms are present, the two or more heteroatoms can all be the same as one another, or some or all of the two or more heteroatoms can each be different from the others. 0062. As used herein, the term “ring refers to any covalently closed structure. Rings include, for example, car bocycles (e.g., aryls and cycloalkyls), heterocycles (e.g., het eroaryls and non-aromatic heterocycles), aromatics (e.g. aryls and heteroaryls), and non-aromatics (e.g., cycloalkyls and non-aromatic heterocycles). Rings can be optionally Sub stituted. Rings can be monocyclic or polycyclic. 0063 As used herein, the term “ring system” refers to one, or more than one ring. 0064. As used herein, the term “non-aromatic hetero cycle”, “heterocycloalkyl or "heteroalicyclic” refers to a non-aromatic ring wherein one or more atoms forming the ring is a heteroatom. A “non-aromatic heterocycle' or “het erocycloalkyl group refers to a cycloalkyl group that includes at least one heteroatom selected from nitrogen, oxy gen and Sulfur. The radicals may be fused with an aryl or heteroaryl. Heterocycloalkyl rings can be formed by three, and the like. Depending on the structure, an cycloalkyl group four, five, six, seven, eight, nine, or more than nine atoms. can be a monoradical or a diradical (e.g., an cycloalkylene Heterocycloalkyl rings can be optionally Substituted. In cer group). tain embodiments, non-aromatic heterocycles contain one or 0056 “Cycloalkylalkyl means an alkyl radical, as more carbonyl or thiocarbonyl groups such as, for example, defined herein, substituted with a cycloalkyl group. Non oXo- and thio-containing groups. Examples of heterocy limiting cycloalkylalkyl groups include cyclopropylmethyl, cloalkyls include, but are not limited to, lactams, lactones, cyclobutylmethyl, cyclopentylethyl, cyclohexylmethyl, and cyclic imides, cyclic thioimides, cyclic carbamates, tetrahy the like. drothiopyran, 4H-pyran, tetrahydropyran, piperidine, 1,3-di 0057 The term “ester” refers to a chemical moiety with oxin, 1.3-dioxane, 1,4-dioxin, 1.4-dioxane, piperazine, 1.3- formula —COOR, where R is selected from among alkyl, Oxathiane, 1,4-oxathiin, 1.4-oxathiane, tetrahydro-1,4- cycloalkyl, aryl, heteroaryl (bonded through a ring carbon) thiazine, 2H-1,2-oxazine, maleimide, Succinimide, barbituric and heteroalicyclic (bonded through a ring carbon). Any acid, thiobarbituric acid, dioxopiperazine, hydantoin, dihy hydroxy, or carboxyl side chain on the compounds described drouracil, morpholine, trioxane, hexahydro-1,3,5-triazine, herein can be esterified. The procedures and specific groups tetrahydrothiophene, tetrahydrofuran, pyrroline, pyrrolidine, to make such esters can be found in Greene and Wuts, Pro pyrrolidone, pyrrolidione, pyrazoline, pyrazolidine, imidazo tective Groups in Organic Synthesis, 3rd Ed., John Wiley & line, imidazolidine, 1,3-dioxole, 1,3-dioxolane, 1,3-dithiole, Sons, New York, N.Y., 1999, which is incorporated herein by 1,3-dithiolane, isoxazoline, isoxazolidine, oxazoline, oxazo reference in its entirety. lidine, oxazolidinone, thiazoline, thiazolidine, and 1.3-OX 0058. The term “halo” or, alternatively, “halogen' or athiolane. Illustrative examples of heterocycloalkyl groups, “halide” means fluoro, chloro, bromo or iodo. also referred to as non-aromatic heterocycles, include: 0059. The terms “haloalkyl,” “haloalkenyl,” “haloalky nyl' and “haloalkoxy” include alkyl, alkenyl, alkynyl and alkoxy structures in which at least one hydrogen is replaced O O. O O O O with a halogenatom. In certain embodiments in which two or more hydrogen atoms are replaced with halogen atoms, the S, , N N, N, O, halogen atoms are all the same as one another. In other \ / embodiments in which two or more hydrogen atoms are US 2010/00566.17 A1 Mar. 4, 2010
azetidinyl (derived from azetidine). An example of a 5-mem -continued bered heterocyclic group is thiazolyl. An example of a 6-membered heterocyclic group is pyridyl, and an example of a 10-membered heterocyclic group is quinolinyl. Examples of non-aromatic heterocyclic groups are pyrrolidinyl, tetrahy drofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropy ranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, aZetidi nyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiepa nyl, oxazepinyl, diazepinyl, thiazepinyl, 1.2.3,6-tetrahydro pyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl. 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl. dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo3. 1.0 hexanyl, 3-azabicyclo4.1.0 heptanyl, 3H-indolyl and quinolizinyl. Examples of aromatic heterocyclic groups are pyridinyl, imidazolyl pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl pyrrolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, ben Zofurazanyl, benzothiophenyl, benzothiazolyl, benzox azolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and furo pyridinyl. The foregoing groups, as derived from the groups listed above, may be C-attached or N-attached where such is possible. For instance, a group derived from pyrrole may be pyrrol-1-yl (N-attached) or pyrrol-3-yl (C-attached). Further, a group derived from imidazole may be imidazol-1-yl or the like. The term heteroalicyclic also includes all ring forms imidazol-3-yl (both N-attached) or imidazol-2-yl, imidazol of the carbohydrates, including but not limited to the 4-yl or imidazol-5-yl (all C-attached). The heterocyclic monosaccharides, the disaccharides and the oligosaccha groups include benzo-fused ring systems and ring systems rides. Depending on the structure, a heterocycloalkyl group substituted with one or two oxo (=O) moieties such as pyr can be a monoradical or a diradical (i.e., a heterocycloalky rolidin-2-one. Depending on the structure, a heterocycle lene group). group can be a monoradical or a diradical (i.e., a heterocy clene group). Phosphorous-containing rings include, but are 0065. “Heterocycloalkylalkyl refers to an alkyl group, as not limited to, 1-oxo-phospholanyl, 1-methyl-1-oxo-phos defined herein, substituted with a heterocycloalkyl, as defined phinan-4-yl, 1-phenyl-1-oxo-phosphinan-4-yl, 1-(cyclopro herein. pylmethyl)-1-oxo-phosphinan-4-yl 4-methyl-4-oxo-1.4 0066. The term "heterocycle” refers to heteroaromatic and aZaphosphinan-1-yl 4-phenyl-4-OXO-1,4aZaphosphinan-1- heteroalicyclic groups containing one to four heteroatoms y1, and 4-(cyclopropylmethyl)-4-oxo-1.4aZaphosphinan-1- each selected from O, S and N, wherein each heterocyclic y1. group has from 4 to 10 atoms in its ring system, and with the 0067. The term “membered ring can embrace any cyclic proviso that the ring of said group does not contain two structure. The term “membered' is meant to denote the num adjacent O or S atoms. Herein, whenever the number of ber of skeletal atoms that constitute the ring. Thus, for carbon atoms in a heterocycle is indicated (e.g., C-C het example, cyclohexyl, pyridine, pyran and thiopyran are erocycle), at least one other atom (the heteroatom) must be 6-membered rings and cyclopentyl, pyrrole, furan, and present in the ring. Designations such as "C-C heterocycle thiophene are 5-membered rings. refer only to the number of carbonatoms in the ring and do not refer to the total number of atoms in the ring. It is understood 0068. The term "moiety” refers to a specific segment or that the heterocylic ring can have additional heteroatoms in functional group of a molecule. Chemical moieties are often the ring. Designations such as “4-6 membered heterocycle” recognized chemical entities embedded in or appended to a refer to the total number of atoms that are contained in the ring molecule. (i.e., a four, five, or six membered ring, in which at least one 0069. As used herein, the term “O-carboxy” or “acyloxy” atom is a carbon atom, at least one atom is a heteroatom and refers to a group of formula RC(=O)C)—. the remaining two to four atoms are either carbon atoms or (0070. “Alkylcarbonyloxy” refers to a (alkyl)-C(=O)C)– heteroatoms). In heterocycles that have two or more heteroa group. toms, those two or more heteroatoms can be the same or 0071. As used herein, the term “alkoxycarbonyl refers to different from one another. Heterocycles can be optionally a group of formula—C(=O)CR. Substituted. Binding to a heterocycle can be at a heteroatom or (0072 “Carboxy” means a-C(O)OH radical. via a carbonatom. Non-aromatic heterocyclic groups include 0073. As used herein, the term “acetyl refers to a group of groups having only 4 atoms in their ring system, but aromatic formula—C(=O)CH. heterocyclic groups must have at least 5 atoms in their ring (0074) “Acyl” refers to the group –C(O)R system. The heterocyclic groups include benzo-fused ring (0075. The term “optionally substituted” or “substituted” systems. An example of a 4-membered heterocyclic group is means that the referenced group may be substituted with one US 2010/00566.17 A1 Mar. 4, 2010 or more additional group(s) individually and independently 268. Subsequently, the cultures are pretreated for one hour in selected from alkyl, cycloalkyl, aryl, heteroaryl, heteroalicy the presence of a candidate neuroprotective agent and then clic, hydroxy, alkoxy, aryloxy, alkylthio, arylthio, alkylsul incubated with a cytotoxic agent for about 18 hours. foxide, arylsulfoxide, alkylsulfone, arylsulfone, cyano, halo, Examples of cytotoxic agents include, but are not limited to, carbonyl, isocyanato, thiocyanato, isothiocyanato, nitro, per oxidative stressors (e.g., 3-nitropropionic acid (3-NP) or haloalkyl, fluoroalkyl, silyl, and amino, including mono- and HO), excitatory amino acids (e.g., kainate), or neurotoxic di-Substituted amino groups, and the protected derivatives proteins (e.g., Hiv Tat, or AB peptides). If the survival of cell thereof. By way of example an optional substituents may be cultures in the presence of the candidate compound and the LR, wherein each L is independently selected from a bond, cytotoxic agent (e.g., 3-NP) is significantly greater than that of cultures incubated with the cytotoxic agent alone, the can didate test compound is considered to have neuroprotective activity. The level of protection (“percent protection') pro tuted or unsubstituted C-C alkyl), or -(substituted or unsub vided by the test compound at a given test concentration in stituted C-C alkenyl); and each R is independently selected vitro can be expressed as: from H., (substituted or unsubstituted C-C alkyl), (substi tuted or unsubstituted C-C cycloalkyl), heteroaryl, or het eroalkyl. ( Cell Viability with Cytoxic agent + 0076. The compounds presented herein may possess one Test Compound x 100 or more stereocenters and each center may exist in the R or S (Cell Viability with Cytotoxic agent alone) configuration. The compounds presented herein include all (Control Cell Viability (medium alone) - diastereomeric, enantiomeric, and epimeric forms as well as (Cell Viability with Cytotoxic agent alone) the appropriate mixtures thereof. Stereoisomers, if desired, may be obtained, for example, by the separation of stereoi Somers by chiral chromatographic columns. I0081. In some embodiments, at a concentration of 10 uM, 0077. The methods and formulations described herein a neuroprotective compound described herein, provides at include the use of N-oxides, crystalline forms (also known as least 6% protection against a cytotoxic agent, i.e., at least 7%. polymorphs), or pharmaceutically acceptable salts of com 15%, 18%, 19%, 20%, 23%, 26%, 28%, 32%, 33%, 36%, pounds described herein, as well as active metabolites of 38%. 39%, 42%, 50%, 54%, 56%, 58%, 61%. 65%, 69%, these compounds having the same type of activity. In some 70%, 77%,82%, 84%, 97%, or any other percent from at least situations, compounds may exist as tautomers. All tautomers 6% to 100% protection against the cytotoxic agent. are included within the scope of the compounds presented I0082 In some embodiments, at a concentration of 10 uM, herein. In addition, the compounds described herein can exist a neuroprotective compound described herein, provides in unsolvated as well as Solvated forms with pharmaceutically greater than 100% protection, which signifies that, in addition acceptable solvents such as water, ethanol, and the like. The to blocking cytotoxic agent-induced cell death, the neuropro Solvated forms of the compounds presented herein are also tective compound also inhibits spontanteous cell death in the considered to be disclosed herein. control cell cultures (i.e., cultures exposed to medium alone). In Some embodiments, a neuroprotective compound Neuroprotective Compounds described herein provides, e.g., 101, 128%, 129, or 15.1% 0078. The neuroprotective compounds for use in the phar protection. maceutical compositions and methods described herein are I0083) A variety of methods for determining cell viability compounds of Formula I. Formula II, and/or compounds may be used. In some embodiments, cell viability is assessed found in Tables 2 and 3. Neuroprotective compounds suitable based on the “MTT' assay method described in Mosmann for the methods described herein also can come from a variety (1983), J Immunol Methods, 65(1-2):55-63, or a variant of Sources including both natural (e.g., plant extracts) and thereof. This assay is based on the ability of a mitochondrial synthetic. For example, neuroprotective compounds falling dehydrogenase enzyme from viable cells to cleave the tetra within the class of triterpenes can be extracted from plants of Zolium rings of the pale yellow MTT and form dark blue the order Rutales, e.g., from extracts of seeds, oils, kernels, formazan crystals, which are trapped in cells and readily leaves and bark of various plants from the Meliaceae family, quantified by ELISA spectrophotometry. including Neem and Mahogany. See, e.g., Roy et al. (2006), I0084. High throughput cell viability assays may be used, Biol Pharm Bull, 29(2):191-201. and are particularly useful for screening, with routine effort, 007.9 The neuroprotective compounds described herein a great number of candidate neuroprotective compounds or are identified or characterized in an in vitro cellular assay, structural variants of identified neuroprotective compounds, e.g., an assay that determines the viability of neurons or glia e.g., structural variants of Formula I or Formula II described in the presence of a cytotoxic challenge and one or more herein. See, e.g., J Biomol Screen, 9(6):506-515; Carrier et al. concentrations of a candidate neuroprotective compound. (2006), J Neurosci Methods, 154(1-2):239-244: See, e.g., In Such assays are useful for identifying and testing the in vitro addition, high throughput screening systems are commer neuroprotective potency of candidate neuroprotective com cially available (see, e.g., Zymark Corp., Hoplinton, Mass.; pounds. Air Technical Industries, Mentor, Ohio; Beckman Instru 0080 For example, a cell viability assay can be used in a ments, Inc. Fullerton, Calif.; Precision Systems, Inc., Natick, preliminary screen on a large series of compounds each of Mass., etc.). These systems typically automate entire proce which is tested at a fixed concentration. In some embodi dures including all sample and reagent pipetting, liquid dis ments, mixed hippocampal cell cultures are prepared as pensing, timed incubations, and final readings of the micro described in, e.g., Haughey et al. (1999), J Neurochem, 73(4): plate in detector(s) appropriate for the assay. Automated 1363-1374, and Haughey etal (2004), J Neurosci, 24(1):257 systems thereby allow the identification and characterization
US 2010/00566.17 A1 Mar. 4, 2010
exist as tautomers. All tautomers are included within the of a subject. It is understood that the plasma concentration of Scope of the compounds presented herein. Additionally, the compounds of Formula I or Formula II may vary significantly compounds described herein can exist in unsolvated as well between subjects, due to variability with respect to metabo as Solvated forms with pharmaceutically acceptable solvents lism and/or possible interactions with other therapeutic such as water, ethanol, and the like. The solvated forms of the agents. In accordance with one embodiment disclosed herein, compounds presented herein are also considered to be dis the blood plasma concentration of the compounds of Formula closed herein. I or Formula II may vary from subject to subject Likewise, 0096 “Antioxidants' include, for example, butylated values such as maximum plasma concentration (Cmax) or hydroxytoluene (BHT), sodium ascorbate, ascorbic acid, time to reach maximum plasma concentration (Tmax), or sodium metabisulfite and tocopherol. In certain embodi total area under the plasma concentration time curve (AUC ments, antioxidants enhance chemical stability where (0-oo)) may vary from subject to subject Due to this variabil required. ity, the amount necessary to constitute "a therapeutically 0097. In certain embodiments, compositions provided effective amount of a compound of Formula I or Formula II herein may also include one or more preservatives to inhibit may vary from Subject to Subject. microbial activity. Suitable preservatives include mercury 0102 “Carrier materials” include any commonly used containing Substances such as merfen and thiomersal; stabi excipients in pharmaceutics and should be selected on the lized chlorine dioxide; and quaternary ammonium com basis of compatibility with compounds disclosed herein, Such pounds Such aS benzalkonium chloride, as, compounds of Formula I or Formula II, and the release cetyltrimethylammonium bromide and cetylpyridinium chlo profile properties of the desired dosage form. Exemplary ride. carrier materials include, e.g., binders, Suspending agents, 0098. Formulations described herein may benefit from disintegration agents, filling agents, Surfactants, solubilizers, antioxidants, metal chelating agents, thiol containing com stabilizers, lubricants, wetting agents, diluents, and the like. pounds and other general stabilizing agents. Examples of "Pharmaceutically compatible carrier materials' may Such stabilizing agents, include, but are not limited to: (a) include, but are not limited to, acacia, gelatin, colloidal sili about 0.5% to about 2% w/v glycerol, (b) about 0.1% to about con dioxide, calcium glycerophosphate, calcium lactate, mal 1% w/v methionine, (c) about 0.1% to about 2% w/v mono todextmin, glycerine, magnesium silicate, polyvinylpyrrolli thioglycerol, (d) about 1 mM to about 10 mM EDTA, (e) done (PVP), cholesterol, cholesterolesters, sodium caseinate, about 0.01% to about 2% w/v ascorbic acid, (f) 0.003% to Soy lecithin, taurocholic acid, phosphotidylcholine, sodium about 0.02% w/v polysorbate 80, (g) 0.001% to about 0.05% chloride, tricalcium phosphate, dipotassium phosphate, cel w/v. polysorbate 20, (h) arginine, (i) heparin, () dextran Sul lulose and cellulose conjugates, Sugars sodium Stearoyl lac fate, (k) cyclodextrins, (1) pentosan polysulfate and other tylate, carrageenan, monoglyceride, diglyceride, pregelati heparinoids, (m) divalent cations such as magnesium and nized Starch, and the like. See, e.g., Remington: The Science Zinc, or (n) combinations thereof. and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.; Mack 0099 “Binders' impart cohesive qualities and include, Publishing Company, 1995); Hoover, John E., Remington's e.g., alginic acid and salts thereof; cellulose derivatives Such Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa. as carboxymethylcellulose, methylcellulose (e.g., Metho 1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceuti cel(R), hydroxypropylmethylcellulose, hydroxyethylcellu cal Dosage Forms, Marcel Decker, New York, N.Y., 1980; and lose, hydroxypropylcellulose (e.g., Klucel(R), ethylcellulose Pharmaceutical Dosage Forms and Drug Delivery Systems, (e.g., Ethocel(R), and microcrystalline cellulose (e.g., Seventh Ed. (Lippincott Williams & Willins 1999). Avicel(R); microcrystalline dextrose; amylose; magnesium 0103) “dispersing agents.” and/or "viscosity modulating aluminum silicate; polysaccharide acids; bentonites; gelatin: agents' include materials that control the diffusion and polyvinylpyrrolidone/vinyl acetate copolymer, crosspovi homogeneity Qfa drug through liquid media or a granulation done; poVidone; starch; pregelatinized Starch; tragacanth, method or blend method. In some embodiments, these agents dextrin, a Sugar, Such as Sucrose (e.g., Dipac.R.), glucose, also facilitate the effectiveness of a coating or eroding matrix. dextrose, molasses, mannitol, Sorbitol. Xylitol (e.g., Xyl Exemplary diffusion facilitators/dispersing agents include, itabR), and lactose; a natural or synthetic gum Such as acacia, e.g., hydrophilic polymers, electrolytes, Tween R. 60 or 80, tragacanth, ghatti gum, mucilage of isapol husks, polyvi PEG, polyvinylpyrrolidone (PVP; commercially known as nylpyrrolidone (e.g., Polyvidone(R) CL, Kollidon R. CL, Poly Plasdone(R), and the carbohydrate-based dispersing agents plasdone RXL-10), larch arabogalactan, Veegum(R), polyeth Such as, for example, hydroxypropyl celluloses (e.g., HPC. ylene glycol, waxes, sodium alginate, and the like. HPC-SL, and HPC-L), hydroxypropyl methylcelluloses 0100 “Bioavailability” refers to the percentage of the (e.g., HPMC K100, HPMC K4M, HPMC K15M, and HPMC weight of compounds disclosed herein, such as, compounds K10OM), carboxymethylcellulose sodium, methylcellulose, of Formula I or Formula II, that is delivered into the general hydroxyethylcellulose, hydroxypropylcellulose, hydrox circulation of the animal or human being studied. The total ypropylmethylcellulose phthalate, hydroxypropylmethylcel exposure (AUC(O-Oo)) of a drug when administered intrave lulose acetate stearate (HPMCAS), noncrystalline cellulose, nously is usually defined as 100% bioavailable (F%). “Oral magnesium aluminum silicate, triethanolamine, polyvinyl bioavailability” refers to the extent to which compounds dis alcohol CPVA), vinyl pyrrolidone/vinyl acetate copolymer closed herein, such as, compounds of Formula I or Formula II (S630), 4-(1,1,3,3-tetramethylbutyl)-phenol polymer with that are absorbed into the general circulation when the phar ethylene oxide and formaldehyde (also known as tyloxapol), maceutical composition is taken orally as compared to intra poloxamers (e.g., Pluronics F68(R), F88(R), and F1088), which venous injection. are block copolymers of ethylene oxide and propylene 0101) “Blood plasma concentration” refers to the concen oxide); and poloxamines (e.g., Tetronic 908R, also known as tration of compounds disclosed herein, Such as, compounds Poloxamine 908R, which is a tetrafunctional block copoly of Formula I or Formula II, in the plasma component of blood mer derived from sequential addition of propylene oxide and US 2010/00566.17 A1 Mar. 4, 2010
ethylene oxide to ethylenediamine (BASF Corporation, Par aluminum silicate), a gum Such as agar, guar, locust bean, Sippany, N.J.)), polyvinylpyrrolidone K12, polyvinylpyrroli Karaya, pectin, or tragacanth, sodium starch glycolate, ben done K17, polyvinylpyrrolidone K25, or polyvinylpyrroli tonite, a natural Sponge, a Surfactant, a resin Such as a cation done K30, polyvinylpyrrolidone/vinyl acetate copolymer exchange resin, citrus pulp, sodium lauryl Sulfate, sodium (S-630), polyethylene glycol, e.g., the polyethylene glycol lauryl Sulfate in combination starch, and the like. can have a molecular weight of about 300 to about 6000, or 0106 “Drug absorption” or “absorption' typically refers about 3350 to about 4000, or about 7000 to about 5400, to the process of movement of drug from site of administra Sodium carboxymethylcellulose, methylcellulose, polysor tion of a drug across a barrier into a blood vessel or the site of bate-80, Sodium alginate, gums, such as, e.g., gum tragacanth action, e.g., a drug moving from the gastrointestinal tract into and gum acacia, guar gum, Xanthans, including Xanthan gum, the portal vein or lymphatic system. Sugars, cellulosics, such as, e.g., sodium carboxymethylcel 0.107 The term “effective amount,” as used herein, refers lulose, methylcellulose, sodium carboxymethylcellulose, to a Sufficient amount of an agent or a compound being polysorbate-80, Sodium alginate, polyethoxylated Sorbitan administered which will relieve to some extent one or more of monolaurate, polyethoxylated Sorbitan monolaurate, povi the symptoms of the disease or condition being treated. The done, carbomers, polyvinyl alcohol (PVA), alginates, chito result can be reduction and/or alleviation of the signs, symp sans and combinations thereof. Plasticizcers such as cellulose toms, or causes of a disease, or any other desired alteration of or triethyl cellulose can also be used as dispersing agents. a biological system. An appropriate 'effective' amount in any Dispersing agents particularly useful in liposomal disper individual case may be determined using techniques, such as sions and self-emulsifying dispersions are dimyristoyl phos a dose escalation study. phatidyl choline, natural phosphatidyl choline from eggs, 0108. An "enteric coating is a substance that remains natural phosphatidylglycerol from eggs, cholesterol and iso Substantially intact in the stomach but dissolves and releases propyl myristate. the drug in the Small intestine or colon. Generally, the enteric 0104. The term "diluenf refers to chemical compounds coating comprises a polymeric material that prevents release that are used to dilute the compound of interest prior to in the low pH environment of the stomach but that ionizes at delivery. Diluents can also be used to stabilize compounds a higher pH, typically a pH of 6 to 7, and thus dissolves because they can provide a more stable environment. Salts sufficiently in the small intestine or colon to release the active dissolved in buffered solutions (which also can provide pH agent therein. control or maintenance) are utilized as diluents, including, 0109 "Filling agents' include compounds such as lactose, but not limited to a phosphate buffered saline solution. In calcium carbonate, calcium phosphate, dibasic calcium phos certain embodiments, diluents increase bulk of the composi phate, calcium Sulfate, microcrystalline cellulose, cellulose tion to facilitate compression or create sufficient bulk for powder, dextrose, dextrates, dextran, starches, pregelatinized homogenous blend for capsule filling. Such compounds starch, Sucrose, Xylitol, lactitol, mannitol, Sorbitol, Sodium include e.g., lactose, starch, mannitol, Sorbitol, dextrose, chloride, polyethylene glycol, and the like. microcrystalline cellulose such as Avicel(R: dibasic calcium 0110 "Flavoring agents' and/or “sweeteners' useful in phosphate, dicalcium phosphate dihydrate; tricalcium phos the formulations described herein, include, e.g., acacia syrup, phate, calcium phosphate; anhydrous lactose, spray-dried acesulfame K, alitame, anise, apple, aspartame, banana, lactose; pregelatinized starch, compressible Sugar, Such as Bavarian cream, berry, black currant, butterscotch, calcium Di-PacR (Amstar); mannitol, hydroxypropylmethylcellu citrate, camphor, caramel, cherry, cherry cream, chocolate, lose, hydroxypropylmethylcellulose acetate Stearate, cinnamon, bubble gum, citrus, citrus punch, citrus cream, Sucrose-based diluents, confectioner's Sugar, monobasic cal cotton candy, cocoa, cola, cool cherry, cool citrus, cyclamate, cium sulfate monohydrate, calcium Sulfate dihydrate; cal cylamate, dextrose, eucalyptus, eugenol, fructose, fruit cium lactate trihydrate, dextrates; hydrolyzed cereal Solids, punch, ginger, glycyrrhetinate, glycyrrhiza (licorice) syrup, amylose; powdered cellulose, calcium carbonate; glycine, grape, grapefruit, honey, isomalt, lemon, lime, lemon cream, kaolin; mannitol, Sodium chloride; inositol, bentonite, and monoammonium glyrrhizinate (MagnaSweet(R), maltol, the like. mannitol, maple, marshmallow, menthol, mint cream, mixed 0105. The term “disintegrate” includes both the dissolu berry, neohesperidine DC, neotame, orange, pear, peach, pep tion and dispersion of the dosage form when contacted with permint, peppermint cream, ProSweet(R) Powder, raspberry, gastrointestinal fluid. "Disintegration agents or disinte root beer, rum, saccharin, Safrole, Sorbitol, spearmint, spear grants' facilitate the breakup or disintegration of a Substance. mint cream, Strawberry, Strawberry cream, Stevia, Sucralose, Examples of disintegration agents include a starch, e.g., a Sucrose, Sodium saccharin, saccharin, aspartame, acesulfame natural starch Such as corn starch or potato starch, a pregela potassium, mannitol, talin, Sylitol. Sucralose, Sorbitol, Swiss tinized starch such as National 1551 or Amijel R., or sodium cream, tagatose, tangerine, thaumatin, tutti fruitti, Vanilla, starch glycolate such as Promogel(R) or Explotab(R), a cellu walnut, watermelon, wild cherry, wintergreen, xylitol, or any lose Such as a wood product, methylcrystalline cellulose, e.g., combination of these flavoring ingredients, e.g., anise-men Avice1(R), Avice1(R) PH101, Avice1(R) PH102, Avicel(R) PH105, thol, cherry-anise, cinnamon-orange, cherry-cinnamon, Elcema(RDP100, Emcocel(RD, Vivacel(R), Ming Tia(RD, and chocolate-mint, honey-lemon, lemon-lime, lemon-mint, Solka-FlocR), methylcellulose, croscarmellose, or a cross menthol-eucalyptus, orange-cream, Vanlla-mint, and mix linked cellulose, such as cross-linked sodium carboxymeth tures thereof. ylcellulose (Ac-Di-SolR), cross-linked carboxymethylcellu 0111 “Lubricants' and "glidants' are compounds that lose, or cross-linked croScarmellose, a cross-linked starch prevent, reduce or inhibit adhesion or friction of materials. Such as Sodium starch glycolate, a cross-linked polymer Such Exemplary lubricants include, e.g., Stearic acid, calcium as crosspoVidone, a cross-linked polyvinylpyrrolidone, algi hydroxide, talc, Sodium Stearyl fumerate, a hydrocarbon Such nate such as alginic acid or a salt of alginic acid Such as as mineral oil, or hydrogenated vegetable oil such as hydro Sodium alginate, a clay Such as Veegum R. HV (magnesium genated soybean oil (SteroteXCR), higher fatty acids and their US 2010/00566.17 A1 Mar. 4, 2010
alkali-metal and alkaline earth metal salts, such as aluminum, 0.122 The term “subject as used herein, refers to an ani calcium, magnesium, zinc, Stearic acid, sodium Stearates, mal which is the object of treatment, observation or experi glycerol, talc, waxes, Stearowet(R), boric acid, Sodium ben ment By way of example only, a Subject may be, but is not Zoate, sodium acetate, sodium chloride, leucine, a polyethyl limited to, a mammal including, but not limited to, a human. ene glycol (e.g., PEG4000) or a methoxypolyethylene glycol 0123 "Suspending agents' include compounds such as such as CarbowaxTM, sodium oleate, sodium benzoate, glyc polyvinylpyrrolidone, e.g., polyvinylpyrrolidone K12, poly eryl behenate, polyethylene glycol, magnesium or sodium vinylpyrrolidone K17, polyvinylpyrrolidone K25, or polyvi lauryl sulfate, colloidal silica such as SyloidTM, Cab-O-Sil.R., nylpyrrolidone K30, vinyl pyrrolidone/vinyl acetate copoly a starch Such as corn starch, silicone oil, a Surfactant, and the mer (S630), polyethylene glycol, e.g., the polyethylene like. glycol can have a molecular weight of about 300 to about 0112 A “measurable serum concentration” or “measur 6000, or about 3350 to about 4000, or about 7000 to about able plasma concentration” describes the blood serum or 5400, sodium carboxymethylcellulose, methylcellulose, blood plasma concentration, typically measured in mg, ug, or hydroxypropylmethylcellulose, hydroxymethylcellulose ng of therapeutic agent per ml, dl, or 1 of blood serum, acetate stearate, polysorbate-80, hydroxyethylcellulose, absorbed into the bloodstream after administration. As used Sodium alginate, gums, such as, e.g., gum tragacanthandgum herein, measurable plasma concentrations are typically mea acacia, guar gum, Xanthans, including Xanthan gum, Sugars, Sured in ng/ml org/ml. cellulosics, such as, e.g., sodium carboxymethylcellulose, 0113. The terms “neuroprotective.” “neuroprotection,” or methylcellulose, sodium carboxymethylcellulose, hydrox “neuroprotectant as used herein refer to the ability of an ypropylmethylcellulose, hydroxyethylcellulose, polysor agent (e.g., a compound described herein) to significantly bate-80, Sodium alginate, polyethoxylated Sorbitan monolau prevent or reduce the occurrence of spontaneous or induced rate, polyethoxylated Sorbitan monolaurate, povidone and the death (e.g., by apoptosis) of neurons or glia in vitro or in vivo, like. relative to the ability of a control reagent (e.g., cell culture 0.124 “Surfactants’ include compounds such as sodium medium, or a drug vehicle such as DMSO). lauryl sulfate, sodium docusate, Tween 60 or 80, triacetin, 0114. A "prognostic biomarker, as referred to herein, is vitamin ETPGS, sorbitan monooleate, polyoxyethylene sor any molecular, biochemical, metabolic, cellular, or structural bitan monooleate, polysorbates, polaxomers, bile salts, glyc entity (or a ratio of such entities), the presence or level of eryl monostearate, copolymers of ethylene oxide and propy which relates to a likelihood of developing, suffering from, or lene oxide, e.g., Pluronic R (BASF), and the like. Some other having a relapse of a pathological condition. surfactants include polyoxyethylene fatty acid glycerides and 0115 “Pharmacodynamics' refers to the factors which Vegetable oils, e.g., polyoxyethylene (60) hydrogenated cas determine the biologic response observed relative to the con tor oil; and polyoxyethylene alkylethers and alkylphenyl centration of drug at a site of action. ethers, e.g., octoxynol 10, Octoxynol 40. In some embodi 0116 “Pharmacolinetics' refers to the factors which ments, Surfactants may be included to enhance physical sta determine the attainment and maintenance of the appropriate bility or for other purposes. concentration of drug at a site of action. 0.125 “Viscosity enhancing agents' include, e.g., methyl 0117 “Plasticizers' are compounds used to soften the cellulose, Xanthan gum, carboxymethyl cellulose, hydrox microencapsulation material or film coatings to make them ypropyl cellulose, hydroxypropylmethyl cellulose, hydrox less brittle. Suitable plasticizers include, e.g., polyethylene ypropylmethyl cellulose acetate Stearate, hydroxypropylm glycols such as PEG 300, PEG 400, PEG 600, PEG 1450, ethyl cellulose phthalate, carbomer, polyvinyl alcohol, PEG 3350, and PEG 800, stearic acid, propylene glycol, oleic alginates, acacia, chitosans and combinations thereof. acid, triethyl cellulose and triacetin. In some embodiments, 0.126 “Wetting agents’ include compounds such as oleic plasticizers can also function as dispersing agents or wetting acid, glyceryl monostearate, Sorbitan monooleate, Sorbitan agents. monolaurate, triethanolamine oleate, polyoxyethylene Sorbi 0118. The term “prophylactically effective amount, as tan monooleate, polyoxyethylene Sorbitan monolaurate, used herein, refers that amount of a composition applied to a Sodium docusate, Sodium oleate, Sodium lauryl Sulfate, patient which will relieve to some extent one or more of the sodium doccusate, triacetin, Tween 80, vitamin E TPGS, symptoms of a disease, condition or disorder being treated. In ammonium salts and the like. Such prophylactic applications, such amounts may depend on I0127. The compositions described herein can be formu the patient's state of health, weight, and the like. lated for administration to a subject via any means including, 0119 “Solubilizers” include compounds such as triacetin, but not limited to, oral, parenteral (e.g., intravenous, Subcu triethylcitrate, ethyl oleate, ethyl caprylate, sodium lauryl taneous, or intramuscular), buccal, intranasal, rectal or trans sulfate, sodium doccusate, vitamin E TPGS, dimethylaceta dermal administration routes. As used herein, the term "sub mide, N-methylpyrrolidone, N-hydroxyethylpyrrolidone, ject' is used to mean an animal, preferably a mammal, polyvinylpyrrolidone, hydroxypropylmethyl cellulose, including a human or non-human. The terms patient and hydroxypropyl cyclodextrins, ethanol, n-butanol, isopropyl Subject may be used interchangeably. alcohol, cholesterol, bile salts, polyethylene glycol 200-600, I0128 Moreover, the pharmaceutical compositions glycofurol, transcutol, propylene glycol, and dimethyl isos described herein, which include a compound of Formula I or orbide and the like. Formula II, can be formulated into any Suitable dosage form, 0120 "Stabilizers’ include compounds such as any anti including but not limited to, aqueous oral dispersions, liquids, oxidation agents, buffers, acids, preservatives and the like. gels, syrups, elixirs, slurries, Suspensions and the like, for oral 0121 "Steady state.” as used herein, is when the amount of ingestion by a patient to be treated, Solid oral dosage forms, drug administered is equal to the amount of drug eliminated aerosols, controlled release formulations, fast melt formula within one dosing interval resulting in a plateau or constant tions, effervescent formulations, lyophilized formulations, plasma drug exposure. tablets, powders, pills, dragees, capsules, delayed release for US 2010/00566.17 A1 Mar. 4, 2010
mulations, extended release formulations, pulsatile release tions as homogeneous, it is meant that the particles of the formulations, multiparticulate formulations, and mixed compound of Formula I or Formula II, are dispersed evenly immediate release and controlled release formulations. throughout the composition so that the composition may be 0129. Pharmaceutical preparations for oral use can be readily subdivided into equally effective unit dosage forms, obtained by mixing one or more solid excipient with one or Such as tablets, pills, and capsules. The individual unit dos more of the compounds described herein, optionally grinding ages may also include film coatings, which disintegrate upon the resulting mixture, and processing the mixture of granules, oral ingestion or upon contact with diluent. These formula after adding suitable auxiliaries, if desired, to obtaintablets or tions can be manufactured by pharmacological techniques. dragee cores. Suitable excipients include, for example, fillers 0.134. In another aspect, dosage forms may include Such as Sugars, including lactose, Sucrose, mannitol, or sbrbi microencapsulated formulations. In some embodiments, one tol; cellulose preparations such as, for example, maize starch, or more other compatible materials are present in the wheat starch, rice starch, potato starch, gelatin, gum traga microencapsulation material. Exemplary materials include, cinth, methylcellulose, microcrystalline cellulose, hydrox but are not limited to, pH modifiers, erosion facilitators, anti ypropylmethylcellulose, Sodium carboxymethylcellulose; or foaming agents, antioxidants, flavoring agents, and carrier others such as: polyvinylpyrrolidone (PVP or povidone) or materials such as binders, Suspending agents, disintegration calcium phosphate. If desired, disintegrating agents may be agents, filling agents, Surfactants, solubilizers, stabilizers, added, such as the cross linked croScarmellose sodium, poly lubricants, wetting agents, and diluents. vinylpyrrolidone, agar, oralginic acid or a salt thereof such as 0.135 Materials useful for the microencapsulation Sodium alginate. described herein include materials compatible with com 0130 Dragee cores are provided with suitable coatings. pounds of Formula I or Formula II, which sufficiently isolate For this purpose, concentrated Sugar Solutions may be used, the compound of Formula I or Formula II from other non which may optionally contain gum arabic, talc, polyvinylpyr compatible excipients. Materials compatible with com rolidone, carbopol gel, polyethylene glycol, and/or titanium pounds of Formula I or Formula II are those that delay the dioxide, lacquer Solutions, and Suitable organic solvents or release of the compounds of Formula I or Formula II in vivo. solvent mixtures. Dyestuffs or pigments may be added to the 0.136 Exemplary microencapsulation materials useful for tablets or dragee coatings for identification or to characterize delaying the release of the formulations including com different combinations of active compound doses. pounds of Formula I or Formula II, include, but are not limited 0131 Pharmaceutical preparations which can be used to, hydroxypropyl cellulose ethers (RPC) such as Klucel(R) or orally include push fit capsules made of gelatin, as well as Nisso HPC, low-substituted hydroxypropyl cellulose ethers soft, sealed capsules made of gelatin and a plasticizer, such as (L-HPC), hydroxypropyl methyl cellulose ethers (IPMC) glycerol or Sorbitol. The push fit capsules can contain the such as Seppi film-LC, Pharmacoat(R&, Metolose SR, Metho active ingredients in admixture with filler such as lactose, cel(R)-E, OpadryYS, PrimaFlo, Benecel MP824, and Benecel binders such as starches, and/or lubricants such as talc or MP843, methylcellulose polymers such as Methocel(R)-A magnesium Stearate and, optionally, stabilizers. In soft cap hydroxypropylmethylcellulose acetate stearate Aqoat (HF Sules, the active compounds may be dissolved or Suspended in LS, HF-LG, HF-MS) and Metolose(R), Ethylcelluloses (EC) Suitable liquids, such as fatty oils, liquid paraffin, or liquid and mixtures thereof such as E461, Ethocel(R), Aqualon(R)-EC, polyethylene glycols. In addition, stabilizers may be added. Surelease R, Polyvinyl alcohol (PVA) such as Opadry AMB, All formulations for oral administration should be in dosages hydroxyethylcelluloses such as Natrosol(R), carboxymethyl Suitable for Such administration. celluloses and salts of carboxymethylcelluloses (CMC) such 0.132. In some embodiments, the solid dosage forms dis as Aqualon(R)-CMC, polyvinyl alcohol and polyethylene gly closed herein may be in the form of a tablet, (including a col co-polymers such as Kollicoat IRR, monoglycerides Suspension tablet, a fast-melt tablet, a bite-disintegration tab (Myverol), triglycerides (KLX), polyethylene glycols, modi let, a rapid-disintegration tablet, an effervescent tablet, or a fied food starch, acrylic polymers and mixtures of acrylic caplet), a pill, a powder (including a sterile packaged powder, polymers with cellulose ethers such as EudragitR) EPO, a dispensable powder, or an effervescent powder) a capsule Eudragit R& L30D-55, Eudragit R FS 30D EudragitR) L100 (including both soft or hard capsules, e.g., capsules made 55, Eudragit R& L100, EudragitR) S100, Eudragit R. RD100, from animal-derived gelatin or plant-derived HPMC, or EudragitR) E100, EudragitR) L12.5, EudragitR) S12.5, “sprinkle capsules'), Solid dispersion, Solid solution, bio EudragitRNE30D, and EudragitRNE 40D, cellulose acetate erodible dosage form, controlled release formulations, pulsa phthalate, sepifilms such as mixtures of HPMC and stearic tile release dosage forms, multiparticulate dosage forms, pel acid, cyclodextrins, and mixtures of these materials. lets, granules, or an aerosol. In other embodiments, the 0.137 In still other embodiments, plasticizers such as pharmaceutical formulation is in the form of a powder. In still polyethylene glycols, e.g., PEG 300, PEG 400, PEG 600, other embodiments, the pharmaceutical formulation is in the PEG 1450, PEG 3350, and PEG 800, stearic acid, propylene form of a tablet, including but not limited to, a fast-melt tablet. glycol, oleic acid, and triacetin are incorporated into the Additionally, pharmaceutical formulations of the present microencapsulation material. In other embodiments, the invention may be administered as a single capsule or in mul microencapsulating material useful for delaying the release tiple capsule dosage form. In some embodiments, the phar of the pharmaceutical compositions is from the USP or the maceutical formulation is administered in two, or three, or National Formulary (NF). In yet other embodiments, the four, capsules or tablets. microencapsulation material is Klucel. In still other embodi 0133. In some embodiments, Solid dosage forms, e.g., tab ments, the microencapsulation material is methocel. lets, effervescent tablets, and capsules, are prepared by mix Microencapsulated compounds of Formula I or Formula II ing particles of a compound of Formula I or Formula II, with may be formulated. Such methods include, e.g., spray drying one or more pharmaceutical excipients to form a bulk blend processes, spinning disk-solvent processes, hot melt pro composition. When referring to these bulk blend composi cesses, spray chilling methods, fluidized bed, electrostatic US 2010/00566.17 A1 Mar. 4, 2010 deposition, centrifugal extrusion, rotational Suspension sepa generally predictable location in the intestinal tract more ration, polymerization at liquid-gas or Solid-gas interface, distal to that which would have been accomplished if there pressure extrusion, or spraying solvent extraction bath. In had been no delayed release alterations. In some embodi addition to these, several chemical techniques, e.g., complex ments the method for delay of release is coating. Any coatings coacervation, Solvent evaporation, polymer-polymer incom should be applied to a sufficient thickness such that the entire patibility, interfacial polymerization in liquid media, in situ coating does not dissolve in the gastrointestinal fluids at pH polymerization, in-liquid drying, and desolvation in liquid below about 5, but does dissolve at pH about 5 and above. It media could also be used. Furthermore, other methods such is expected that any anionic polymer exhibiting a pH-depen as roller compaction, extrusion/spheronization, coacervation, dent solubility profile can be used as an enteric coating in the or nanoparticle coating may also be used. practice of the present invention to achieve delivery to the 0.138. In one embodiment, the particles of compounds of lower gastrointestinal tract. In some embodiments the poly Formula I or Formula II are microencapsulated prior to being mers for use in the present invention are anionic carboxylic formulated into one of the above forms. In still another polymers. embodiment, Some or most of the particles are coated prior to 0143. In other embodiments, the formulations described being further formulated by using standard coating proce herein, which include a compound of Formula I or Formula II, dures, such as those described in Remington's Pharmaceuti are delivered using a pulsatile dosage form. A pulsatile dos cal Sciences, 20th Edition (2000). age form is capable of providing one or more immediate 0.139. In other embodiments, the solid dosage formula release pulses at predetermined time points after a controlled tions of the compounds of Formula I or Formula II are plas lag time or at specific sites. Pulsatile dosage forms including ticized (coated) with one or more layers. Illustratively, a plas the formulations described herein, which include a com ticizer is generally a high boiling point solid or liquid. pound of Formula I or Formula II, may be administered using Suitable plasticizers can be added from about 0.01% to about a variety of pulsatile formulations. For example, such formu 50% by weight (w/w) of the coating composition. Plasticizers lations include, but are not limited to, those described in U.S. include, but are not limited to, diethyl phthalate, citrate esters, Pat. Nos. 5,011,692, 5,017,381, 5,229,135, and 5,840,329. polyethylene glycol, glycerol, acetylated glycerides, triace Other pulsatile release dosage forms suitable for use with the tin, polypropylene glycol, polyethylene glycol, triethyl cit present formulations include, but are not limited to, for rate, dibutyl sebacate, Stearic acid, Stearol, Stearate, and castor example, U.S. Pat. Nos. 4,871,549, 5,260,068, 5.260,069, oil. 5,508,040, 5,567,441 and 5,837,284. In one embodiment, the 0140. The pharmaceutical solid oral dosage forms includ controlled release dosage form is pulsatile release solid oral ing formulations described herein, which include a com dosage form including at least two groups of particles, (i.e. pound of Formula I or Formula II, can be further formulated multiparticulate) each containing the formulation described to provide a controlled release of the compound of Formula I herein. The first group of particles provides a substantially or Formula II. Controlled release refers to the release of the immediate dose of the compound of Formula I or Formula II compound of Formula Formula I or Formula II from a dosage upon ingestion by a mammal. The first group of particles can form in which it is incorporated according to a desired profile be either uncoated or include a coating and/or sealant. The over an extended period of time. Controlled release profiles second group of particles includes coated particles, which include, for example, Sustained release, prolonged release, includes from about 2% to about 75%, preferably from about pulsatile release, and delayed release profiles. In contrast to 2.5% to about 70%, and more preferably from about 40% to immediate release compositions, controlled release compo about 70%, by weight of the total dose of the compound of sitions allow delivery of an agent to a subject over an extended Formula I or Formula II in said formulation, in admixture period of time according to a predetermined profile. Such with one or more binders. The coating includes a pharmaceu release rates can provide therapeutically effective levels of tically acceptable ingredient in an amount Sufficient to pro agent for an extended period of time and thereby provide a vide a delay of from about 2 hours to about 7 hours following longer period of pharmacologic response while minimizing ingestion before release of the second dose. Suitable coatings side effects as compared to conventional rapid release dosage include one or more differentially degradable coatings Such forms. Such longer periods of response provide for many as, by way of example only, pH sensitive coatings (enteric inherent benefits that are not achieved with the corresponding coatings) such as acrylic resins (e.g., EudragitR) EPO. short acting, immediate release preparations. EudragitRL30D-55, EudragitRFS30DEudragit RL100-55, 0141. In some embodiments, the solid dosage forms Eudragit R L100, EudragitR) S100, Eudragit R. RD100, described herein can be formulated as enteric coated delayed EudragitR) E100, EudragitR) L12.5, Eudragit RDS12.5, and release oral dosage forms, i.e., as an oral dosage form of a Eudragit R NE30D, Eudragit R NE 40DR) either alone or pharmaceutical composition as described herein which ulti blended with cellulose derivatives, e.g., ethylcellulose, or lizes an enteric coating to affect release in the Small intestine non-enteric coatings having variable thickness to provide of the gastrointestinal tract. The enteric coated dosage form differential release of the formulation that includes a com may be a compressed or molded or extruded tablet/mold pound of Formula I or Formula II. (coated or uncoated) containing granules, powder, pellets, 0144. Additional examples of controlled release delivery beads or particles of the active ingredient and/or other com systems include, e.g., polymer-based systems, such as poly position components, which are themselves coated or lactic and polyglycolic acid, plyanhydrides and polycapro uncoated. The enteric coated oral dosage form may also be a lactone; porous matrices, nonpolymer-based systems that are capsule (coated or uncoated) containing pellets, beads or lipids, including sterols, such as cholesterol, cholesterol granules of the solid carrier or the composition, which are esters and fatty acids, or neutral fats, such as mono-, di- and themselves coated or uncoated. triglycerides; hydrogel release systems; Silastic systems; pep 0142. The term “delayed release' as used herein refers to tide-based systems; wax coatings, bioerodible dosage forms, the delivery so that the release can be accomplished at some compressed tablets using binders and the like. See, e.g., US 2010/00566.17 A1 Mar. 4, 2010
Liberman et al., Pharmaceutical Dosage Forms, 2 Ed., Vol. 1, cellulose ethers (e.g. HPMC K100, HPMC K4M, HPMC pp. 209-214 (1990); Singh et al., Encyclopedia of Pharma K15M, and HPMC K100M), carboxymethylcellulose ceutical Technology, 2nd Ed., pp. 751-753 (2002); U.S. Pat. sodium, methylcellulose, hydroxyethylcellulose, hydrox Nos. 4,327,725, 4,624,848, 4,968,509, 5,461,140, 5,456,923, ypropylmethyl-cellulose phthalate, hydroxypropylmethyl 5,516,527, 5,622,721, 5,686,105, 5,700,410, 5,977,175, cellulose acetate Stearate, noncrystalline cellulose, magne 6,465,014 and 6,932,983. sium aluminum silicate, triethanolamine, polyvinyl alcohol 0145 Liquid formulation dosage forms for oral adminis (PVA), polyvinylpyrrolidone/vinyl acetate copolymer (Plas tration can be aqueous Suspensions selected from the group done(R), e.g., S-630), 4-(1,1,3,3-tetramethylbutyl)-phenol including, but not limited to, pharmaceutically acceptable polymer with ethylene oxide and formaldehyde (also known aqueous oral dispersions, emulsions, Solutions, elixirs, gels, as tyloxapol), poloxamers (e.g., Pluronics F68(R), F88(R), and and syrups. See, e.g., Singh et al., Encyclopedia of Pharma F108(R), which are block copolymers of ethylene oxide and ceutical Technology, 2nd Ed., pp. 754-757 (2002). In addition propylene oxide); and poloxamines (e.g., Tetronic 908R, also to the particles of compound of Formula I or Formula II, the known as Poloxamine 908R, which is a tetrafunctional block liquid dosage forms may include additives, such as: (a) dis copolymer derived from sequential addition of propylene integrating agents; (b) dispersing agents; (c) Wetting agents: oxide and ethylene oxide to ethylenediamine (BASF Corpo (d) at least one preservative, (e) viscosity enhancing agents, ration, Parsippany, N.J.)). In other embodiments, the dispers (f) at least one Sweetening agent, and (g) at least one flavoring ing agent is selected from a group not comprising one of the agent. In some embodiments, the aqueous dispersions can following agents: hydrophilic polymers; electrolytes; further include a crystalline inhibitor. Tween R. 60 or 80; PEG: polyvinylpyrrollidone (PVP): 0146 The aqueous Suspensions and dispersions described hydroxypropylcellulose and hydroxypropyl cellulose ethers herein can remain in a homogenous state, as defined in The (e.g., HPC, HPC-SL, and HPC-L): hydroxypropyl methylcel USP Pharmacists Pharmacopeia (2005 edition, chapter 905), lulose and hydroxypropyl methylcellulose ethers (e.g. HPMC for at least 4 hours. The homogeneity should be determined K100, HPMC K4M, HPMC K15M, BPMC K10OM, and by a sampling method consistent with regard to determining Pharmacoat(R) USP 2910 (Shin-Etsu)); carboxymethylcellu homogeneity of the entire composition. In one embodiment, lose sodium; methylcellulose; hydroxyethylcellulose; an aqueous Suspension can be re-suspended into a homog hydroxypropylmethyl-cellulose phthalate; hydroxypropylm enous Suspension by physical agitation lasting less than 1 ethyl-cellulose acetate Stearate; non-crystalline cellulose; minute. In another embodiment, an aqueous Suspension can magnesium aluminum silicate; triethanolamine; polyvinyl be re-suspended into a homogenous suspension by physical alcohol (PVA): 4-(1,1,3,3-tetramethylbutyl)-phenol polymer agitation lasting less than 45 seconds. In yet another embodi with ethylene oxide and formaldehyde; poloxamers (e.g., ment, an aqueous Suspension can be re-suspended into a Pluronics F68(R), F88(R), and F108(R), which are block copoly homogenous Suspension by physical agitation lasting less mers of ethylene oxide and propylene oxide); or poloxamines than 30 seconds. In still another embodiment, no agitation is (e.g., Tetronic 908(R), also known as Poloxamine 908(R). necessary to maintain a homogeneous aqueous dispersion. 0149 Wetting agents suitable for the aqueous suspensions 0147 Examples of disintegrating agents for use in the and dispersions include, but are not limited to, cetyl alcohol, aqueous Suspensions and dispersions include, but are not glycerol monostearate, polyoxyethylene Sorbitan fatty acid limited to, a starch, e.g., a natural starch Such as corn starch or esters (e.g., the commercially available Tweens(R) Such as e.g., potato starch, a pregelatinized starch Such as National 1551 or Tween 20R) and Tween 80R (ICI Specialty Chemicals)), and Amijel R., or sodium starch glycolate such as Promogel(R) or polyethylene glycols (e.g., Carbowaxs 3350R) and 1450R), Explotab(R); a cellulose such as a wood product, methylcrys and Carbopol 934(R) (Union Carbide)), oleic acid, glyceryl talline cellulose, e.g., Avicel(R), Avicel(R) PHIOI, Avicel(R) monostearate, Sorbitan monooleate, Sorbitan monolaurate, PH102, Avice1(R) PH105, Elcema(R) P100, Emcocel(R), triethanolamine oleate, polyoxyethylene Sorbitan Vivacel(R), Ming TiaR), and Solka-FlocR, methylcellulose, monooleate, polyoxyethylene Sorbitan monolaurate, Sodium croScarmellose, or a cross-linked cellulose, Such as cross oleate, Sodium lauryl Sulfate, sodium docusate, triacetin, Vita linked sodium carboxymethylcellulose (Ac-Di-SolR), cross min E TPGS, sodium taurocholate, simethicone, phosphoti linked carboxymethylcellulose, or cross-linked croScarmel dylcholine and the like lose; a cross-linked starch Such as sodium starch glycolate; a 0150. Suitable preservatives for the aqueous suspensions cross-linked polymer Such as crospovidone; a cross-linked or dispersions described herein include, for example, potas polyvinylpyrrolidone; alginate Such as alginic acid or a salt of sium Sorbate, parabens (e.g., methylparaben and propylpara alginic acid Such as Sodium alginate; a clay Such as Veegum R ben), benzoic acid and its salts, other esters of parahydroxy HV (magnesium aluminum silicate); a gum Such as agar, benzoic acid Such as butylparaben, alcohols such as ethyl guar, locust bean, Karaya, pectin, or tragacanth; sodium alcohol or benzyl alcohol, phenolic compounds such as phe starch glycolate; bentonite; a natural sponge; a Surfactant; a nol, or quaternary compounds such as benzalkonium chlo resin Such as a cation-exchange resin; citrus pulp; sodium ride. Preservatives, as used herein, are incorporated into the lauryl Sulfate; sodium lauryl Sulfate in combination starch; dosage form at a concentration Sufficient to inhibit microbial and the like. growth. 0148. In some embodiments, the dispersing agents Suit 0151. Suitable viscosity enhancing agents for the aqueous able for the aqueous Suspensions and dispersions include, for Suspensions or dispersions described herein include, but are example, hydrophilic polymers, electrolytes, Tween(R) 60 or not limited to, methyl cellulose, Xanthan gum, carboxymethyl 80, PEG, polyvinylpyrrolidone (PVP; commercially known cellulose, hydroxypropyl cellulose, hydroxypropylmethyl as Plasdone(R), and the carbohydrate-based dispersing agents cellulose, Plasdon(R) S-630, carbomer, polyvinyl alcohol, Such as, for example, hydroxypropylcellulose and hydrox alginates, acacia, chitosans and combinations thereof. The ypropyl cellulose ethers (e.g., HPC, HPC-SL, and HPC-L), concentration of the Viscosity enhancing agent will depend hydroxypropyl methylcellulose and hydroxypropyl methyl upon the agent selected and the Viscosity desired. US 2010/00566.17 A1 Mar. 4, 2010
0152 Examples of Sweetening agents suitable for the ingredients. Methods of producing self-emulsifying dosage aqueous Suspensions or dispersions described herein include, forms include, but are not limited to, for example, U.S. Pat. for example, acacia syrup, acesulfame K, alitame, anise, Nos. 5,858,401, 6,667,048, and 6,960,563. apple, aspartame, banana, Bavarian cream, berry, black cur 0.155. It is to be appreciated that there is overlap between rant, butterscotch, calcium citrate, camphor, caramel, cherry, the above-listed additives used in the aqueous dispersions or cherry cream, chocolate, cinnamon, bubblegum, citrus, citrus Suspensions described herein, since a given additive is often punch, citrus cream, cotton candy, cocoa, cola, cool cherry, classified differently by different practitioners in the field, or cool citrus, cyclamate, cylamate, dextrose, eucalyptus, is commonly used for any of several different functions. Thus, eugenol, fructose, fruit punch, ginger, glycyrrhetinate, gly the above-listed additives should be taken as merely exem cyrrhiza (licorice) syrup, grape, grapefruit, honey, isomalt, plary, and not limiting, of the types of additives that can be lemon, lime, lemon cream, monoammonium glyrrhizinate included in formulations described herein. (MagnaSweet(R), maltol, mannitol, maple, marshmallow, 0156 Intranasal formulations of a compound of Formula I menthol, mint cream, mixed berry, neohesperidine DC, or Formula II can be prepared by adapting the methods neotame, orange, pear, peach, peppermint, peppermint described in U.S. Pat. Nos. 4,476,116, 5,116,817 and 6,391, cream, ProSweet(R) Powder, raspberry, root beer, rum, saccha 452. Formulations that include a compound of Formula I or rin, Safrole, Sorbitol, spearmint, spearmint cream, Strawberry, Formula II, which are prepared according to these and other Strawberry cream, Stevia, Sucralose, Sucrose, Sodium saccha techniques are prepared as solutions in Saline, employing rin, Saccharin, aspartame, acesulfame potassium, mannitol, benzyl alcohol or other suitable preservatives, fluorocarbons, tain, Sucralose, Sorbitol, Swiss cream, tagatose, tangerine, and/or other solubilizing or dispersing agents. See, for thaumatin, tutti fruitti, Vanilla, walnut, watermelon, wild example, Ansel, H. C. et al., Pharmaceutical Dosage Forms cherry, wintergreen, Xylitol, or any combination of these fla and Drug Delivery Systems, Sixth Ed. (1995). Preferably Voring ingredients, e.g., anise-menthol, cherry-anise, cinna these compositions and formulations are prepared with Suit mon-orange, cherry-cinnamon, chocolate-mint, honey able nontoxic pharmaceutically acceptable ingredients. lemon, lemon-lime, lemon-mint, menthol-eucalyptus, These ingredients can be found in REMINGTON: THE SCI orange-cream, Vanilla-mint, and mixtures thereof. In one ENCE-AND PRACTICE OF PHARMACY, 21st edition, embodiment, the aqueous liquid dispersion can comprise a 2005, a standard reference in the field. The choice of suitable Sweetening agent or flavoring agent in a concentration rang carriers is highly dependent upon the exact nature of the nasal ing from about 0.001% to about 1.0% the volume of the dosage form desired, e.g., Solutions, Suspensions, ointments, aqueous dispersion. In another embodiment, the aqueous liq or gels. Nasal dosage forms generally contain large amounts uid dispersion can comprise a Sweetening agent or flavoring of water in addition to the active ingredient. Minor amounts of agent in a concentration ranging from about 0.005% to about other ingredients such as pH adjusters, emulsifiers or dispers 0.5% the volume of the aqueous dispersion. In yet another ing agents, preservatives, Surfactants, gelling agents, or buff embodiment, the aqueous liquid dispersion can comprise a ering and other stabilizing and Solubilizing agents may also Sweetening agent or flavoring agent in a concentration rang be present. Preferably, the nasal dosage form should be iso ing from about 0.01% to about 1.0% the volume of the aque tonic with nasal secretions. ous dispersion. 0157 For administration by inhalation, the compounds of 0153. In addition to the additives listed above, the liquid Formula Formula I or Formula II described herein may be in formulations can also include inert diluents such as water or a form as an aerosol, a mist or a powder. Pharmaceutical other solvents, solubilizing agents, and emulsifiers. Exem compositions described herein are conveniently delivered in plary emulsifiers are ethyl alcohol, isopropyl alcohol, ethyl the form of an aerosol spray presentation from pressurized carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, packs or a nebuliser, with the use of a Suitable propellant, e.g., propyleneglycol. 1,3-butyleneglycol, dimethylformamide, dichlorodifluoromethane, trichlorofluoromethane, dichlo Sodium lauryl Sulfate, sodium doccusate, cholesterol, choles rotetrafluoroethane, carbon dioxide or other suitable gas. In terol esters, taurocholic acid, phosphotidylcholine, oils. Such the case of a pressurized aerosol, the dosage unit may be as cottonseed oil, groundnut oil, corn germ oil, olive oil, determined by providing a valve to deliver a metered amount. castor oil, and sesame oil, glycerol, tetrahydrofurfuryl alco Capsules and cartridges of Such as, by way of example only, hol, polyethylene glycols, fatty acid esters of Sorbitan, or gelatin for use in an inhaler or insufflator may be formulated mixtures of these substances, and the like. containing a powder mix of the compound described herein 0154) In some embodiments, the pharmaceutical formula and a suitable powder base Such as lactose or starch tions described herein can be self-emulsifying drug delivery 0158 Buccal formulations that include compounds of systems (SEDDS). Emulsions are dispersions of one immis Formula I or Formula II may be administered using a variety cible phase in another, usually in the form of droplets. Gen of formulations. For example, such formulations include, but erally, emulsions are created by vigorous mechanical disper are not limited to, U.S. Pat. Nos. 4,229,447, 4,596,795, 4,755, Sion. SEDDS, as opposed to emulsions or microemulsions, 386, and 5,739,136. In addition, the buccal dosage forms spontaneously form emulsions when added to an excess of described herein can further include a bioerodible (hydrolys water without any external mechanical dispersion or agita able) polymeric carrier that also serves to adhere the dosage tion. An advantage of SEDDS is that only gentle mixing is form to the buccal mucosa. The buccal dosage form is fabri required to distribute the droplets throughout the solution. cated so as to erode gradually over a predetermined time Additionally, water or the aqueous phase can be added just period, wherein the delivery of the compound of Formula I or prior to administration, which ensures stability of an unstable Formula II, is provided essentially throughout. Buccal drug or hydrophobic active ingredient. Thus, the SEDDS provides delivery avoids the disadvantages encountered with oral drug an effective delivery system for oral and parenteral delivery of administration, e.g., slow absorption, degradation of the hydrophobic active ingredients. SEDDS may provide active agent by fluids present in the gastrointestinal tract improvements in the bioavailability of hydrophobic active and/or first-pass inactivation in the liver. With regard to the US 2010/00566.17 A1 Mar. 4, 2010
bioerodible (hydrolysable) polymeric carrier, it will be appre 0162 Formulations that include a compound of Formula I ciated that virtually any Such carrier can be used, so long as or Formula II, Suitable for intramuscular, Subcutaneous, or the desired drug release profile is not compromised, and the intravenous injection may include physiologically acceptable carrier is compatible with the compound of Formula I or sterile aqueous or non-aqueous solutions, dispersions, Sus Formula II, and any other components that may be present in pensions or emulsions, and sterile powders for reconstitution the buccal dosage unit. Generally, the polymeric carrier com into sterile injectable solutions or dispersions. Examples of prises hydrophilic (water-soluble and water-swellable) poly Suitable aqueous and non-aqueous carriers, diluents, Sol mers that adhere to the wet surface of the buccal mucosa. vents, or vehicles including water, ethanol, polyols (propyle Examples of polymeric carriers useful herein include acrylic neglycol, polyethylene-glycol, glycerol, cremophor and the acid polymers and co, e.g., those known as “carbomers' like), suitable mixtures thereof, vegetable oils (such as olive (Carbopol R, which may be obtained from B.F. Goodrich, is oil) and injectable organic esters such as ethyl oleate. Proper one such polymer). Other components may also be incorpo fluidity can be maintained, for example, by the use of a rated into the buccal dosage forms described herein include, coating such as lecithin, by the maintenance of the required but are not limited to, disintegrants, diluents, binders, lubri particle size in the case of dispersions, and by the use of cants, flavoring, colorants, preservatives, and the like. For Surfactants. Formulations suitable for Subcutaneous injection buccal or Sublingual administration, the compositions may may also contain additives such as preserving, wetting, emul take the form of tablets, lozenges, or gels. Sifying, and dispensing agents. Prevention of the growth of 0159 Transdermal formulations described herein may be microorganisms can be ensured by various antibacterial and administered using a variety of devices that include, but are antifungal agents, such as parabens, chlorobutanol, phenol, not limited to, U.S. Pat. Nos. 3,598,122, 3,598,123, 3,710, sorbic acid, and the like. It may also be desirable to include 795, 3,731,683, 3,742,951, 3,814,097, 3,921,636, 3,972,995, isotonic agents, such as Sugars, sodium chloride, and the like. 3,993,072, 3,993,073, 3,996,934, 4,031,894, 4,060,084, Prolonged absorption of the injectable pharmaceutical form 4,069,307, 4,077,407, 4,201,211, 4,230,105, 4,292,299, can be brought about by the use of agents delaying absorp 4,292,303, 5,336,168, 5,665,378, 5,837,280, 5,869,090, tion, Such as aluminum monostearate and gelatin. 6,923,983, 6,929,801 and 6,946,144. 0163 For intravenous injections, compounds described 0160 The transdermal dosage forms described herein may herein may be formulated in aqueous solutions, preferably in incorporate certain pharmaceutically acceptable excipients. physiologically compatible buffers such as Hank's solution, In one embodiments, the transdermal formulations described Ringer's solution, or physiological saline buffer. For trans herein include at least three components: (1) a formulation of mucosal administration, penetrants appropriate to the barrier a compound of Formula I or Formula II; (2) a penetration to be permeated are used in the formulation. For other enhancer, and (3) an aqueous adjuvant. In addition, transder parenteral injections, appropriate formulations may include mal formulations can include additional components such as, aqueous or nonaqueous solutions, preferably with physi but not limited to, gelling agents, creams and ointment bases, ologically compatible buffers or excipients. and the like. In some embodiments, the transdermal formu 0164 Parenteral injections may involve bolus injection or lation can further include a woven or non-woven backing continuous infusion. Formulations for injection may be pre material to enhance absorption and prevent the removal of the sented in unit dosage form, e.g., in ampoules or in multi-dose transdermal formulation from the skin. In other embodi containers, with an added preservative. The pharmaceutical ments, the transdermal formulations described herein can composition described herein may be in a form suitable for maintain a saturated or Supersaturated State to promote diffu parenteral injection as a sterile Suspensions, solutions or sion into the skin. emulsions in oily or aqueous vehicles, and may contain for 0161 Formulations suitable for transdermal administra mulatory agents such as Suspending, stabilizing and/or dis tion of compounds described herein may employ transdermal persing agents. Pharmaceutical formulations for parenteral delivery devices and transdermal delivery patches and can be administration include aqueous solutions of the active com lipophilic emulsions or buffered, aqueous Solutions, dis pounds in water-soluble form. Additionally, Suspensions of Solved and/or dispersed in a polymer or an adhesive. Such the active compounds may be prepared as appropriate oily patches may be constructed for continuous, pulsatile, or on injection Suspensions. Suitable lipophilic solvents or vehicles demand delivery of pharmaceutical agents. Still further, include fatty oils such as sesame oil, or synthetic fatty acid transdermal delivery of the compounds described herein can esters, such as ethyl oleate or triglycerides, or liposomes. be accomplished by means of iontophoretic patches and the Aqueous injection Suspensions may contain Substances like. Additionally, transdermal patches can provide con which increase the viscosity of the Suspension, such as trolled delivery of the compounds of Formula I or Formula II. sodium carboxymethyl cellulose, sorbitol, or dextran. The rate of absorption can be slowed by using rate-controlling Optionally, the Suspension may also contain Suitable stabiliz membranes or by trapping the compound within a polymer ers or agents which increase the Solubility of the compounds matrix or gel. Conversely, absorption enhancers can be used to allow for the preparation of highly concentrated Solutions. to increase absorption. An absorption enhancer or carrier can Alternatively, the active ingredient may be in powderform for include absorbable pharmaceutically acceptable solvents to constitution with a suitable vehicle, e.g., sterile pyrogen-free assist passage through the skin. For example, transdermal water, before use. devices are in the form of a bandage comprising a backing 0.165. In certain embodiments, delivery systems for phar member, a reservoir containing the compound optionally maceutical compounds may be employed, such as, for with carriers, optionally a rate controlling barrier to deliver example, liposomes and emulsions. In certain embodiments, the compound to the skin of the host at a controlled and compositions provided herein can also include an mucoadhe predetermined rate over a prolonged period of time, and sive polymer, selected from among, for example, carboxym means to secure the device to the skin. ethylcellulose, carbomer (acrylic acid polymer), poly(meth US 2010/00566.17 A1 Mar. 4, 2010 20 ylmethacrylate), polyacrylamide, polycarbophil, acrylic of the patient's life in order to ameliorate or otherwise control acid/butyl acrylate copolymer, sodium alginate and dextran. or limit the symptoms of the patient's disease or condition. 0166 In some embodiments, the compounds described 0173. In the case wherein the patient's status does herein may be administered topically and can be formulated improve, upon the doctor's discretion the administration of into a variety of topically administrable compositions, such as the compounds may be given continuously; alternatively, the Solutions, Suspensions, lotions, gels, pastes, medicated Sticks, dose of drug being administered may be temporarily reduced balms, creams or ointments. Such pharmaceutical com or temporarily Suspended for a certain length of time (i.e., a pounds can contain solubilizers, stabilizers, tonicity enhanc “drug holiday'). The length of the drug holiday can vary ing agents, buffers and preservatives. between 2 days and 1 year, including by way of example only, 0167. The compounds described herein may also be for 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, mulated in rectal compositions such as enemas, rectal gels, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 rectal foams, rectal aerosols, Suppositories, jelly supposito days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 ries, or retention enemas, containing Suppository bases Such days, 300 days, 320 days, 350 days, or 365 days. The dose as cocoa butter or other glycerides, as well as synthetic poly reduction during a drug holiday may be from 10%-100%, mers such as polyvinylpyrrolidone, PEG, and the like. In including, by way of example only, 10%, 15%, 20%, 25%, Suppository forms of the compositions, a low-melting wax 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, Such as, but not limited to, a mixture of fatty acid glycerides, 80%, 85%, 90%, 95%, or 100%. optionally in combination with cocoa butter is first melted. 0.174. Once improvement of the patient's conditions has 0168. In other embodiments, the formulations described occurred, a maintenance dose is administered if necessary. herein, which include a compound of Formula I or Formula II, Subsequently, the dosage or the frequency of administration, are blood brain barrier-permeable (BBB-permeable) nano or both, can be reduced, as a function of the symptoms, to a particle formulations. Methods of producing such BBB-per level at which the improved disease, disorder or condition is meable nanoparticle formulations include, but are not limited retained. Patients can, however, require intermittent treat to, for example, U.S. Pat. Nos. 6,117,454 and 7,025,991. ment on a long-term basis upon any recurrence of symptoms. 0.175. The amount of a given agent that will correspond to Examples of Methods of Dosing and Treatment Regimens Such an amount will vary depending upon factors such as the particular compound, disease or condition and its severity, the 0169. The compounds described herein can be used in the identity (e.g., weight) of the Subject or host in need of treat preparation of medicaments for the treatment of neurodegen ment, but can nevertheless be determined according to the erative diseases or conditions that would benefit, at least in particular circumstances Surrounding the case, including, part, from neuroprotection. In addition, a method for treating e.g., the specific agent being administered, the route of any of the diseases or conditions described herein in a subject administration, the condition being treated, and the Subject or in need of Such treatment, involves administration of phar host being treated. In general, however, doses employed for maceutical compositions containing at least one compound of adult human treatment will typically be in the range of 0.02 Formula I or Formula II described herein, or a pharmaceuti 5000 mg per day, preferably 1-1500 mg per day. The desired cally acceptable salt, pharmaceutically acceptable N-oxide, dose may conveniently be presented in a single dose or as pharmaceutically active metabolite, pharmaceutically divided doses administered simultaneously (or over a short acceptable prodrug, or pharmaceutically acceptable Solvate period of time) or at appropriate intervals, for example as two, thereof, in therapeutically effective amounts to said subject. three, four or more Sub-doses per day. 0170 The compositions containing the compound(s) 0176 The pharmaceutical composition described herein described herein can be administered for prophylactic and/or may be in unit dosage forms suitable for single administration therapeutic treatments. In therapeutic applications, the com of precise dosages. In unit dosage form, the formulation is positions are administered to a patient already Suffering from divided into unit doses containing appropriate quantities of a disease or condition, in an amount Sufficient to cure or at one or more compound. The unit dosage may be in the form least partially arrest the symptoms of the disease or condition. of a package containing discrete quantities of the formulation. Amounts effective for this use will depend on the severity and Non-limiting examples are packaged tablets or capsules, and course of the disease or condition, previous therapy, the powders in vials or ampoules. Aqueous Suspension composi patient's health status, weight, and response to the drugs, and tions can be packaged in single-dose non-reclosable contain the judgment of the treating physician ers. Alternatively, multiple-dose reclosable containers can be 0171 In prophylactic applications, compositions contain used, in which case it is typical to include a preservative in the ing the compounds described herein are administered to a composition. By way of example only, formulations for patient Susceptible to or otherwise at risk of a particular parenteral injection may be presented in unit dosage form, disease, disorder or condition. Such an amount is defined to which include, but are not limited to ampoules, or in multi be a “prophylactically effective amount or dose.” In this use, dose containers, with an added preservative. the precise amounts also depend on the patient's state of 0177. The daily dosages appropriate for the compounds health, weight, and the like. When used in a patient, effective described herein described herein are from about 0.01 to 2.5 amounts for this use will depend on the severity and course of mg/kg per body weight. An indicated daily dosage in the the disease, disorder or condition, previous therapy, the larger mammal, including, but not limited to, humans, is in patient's health status and response to the drugs, and the the range from about 0.5 mg to about 100 mg, conveniently judgment of the treating physician administered in divided doses, including, but not limited to, 0172. In the case wherein the patient's condition does not up to four times a day or in extended release form. Suitable improve, upon the doctor's discretion the administration of unit dosage forms for oral administration include from about the compounds may be administered chronically, that is, for 1 to 50mg active ingredient. The foregoing ranges are merely an extended period of time, including throughout the duration Suggestive, as the number of variables in regard to an indi US 2010/00566.17 A1 Mar. 4, 2010
vidual treatment regime is large, and considerable excursions 0182. In any case, the multiple therapeutic agents (one of from these recommended values are not uncommon. Such which is a compound of Formula I or Formula II described dosages may be altered depending on a number of variables, herein) may be administered in any order, or even simulta not limited to the activity of the compound used, the disease neously. If simultaneously, the multiple therapeutic agents or condition to be treated, the mode of administration, the may be provided in a single, unified form, or in multiple forms requirements of the individual subject, the severity of the (by way of example only, either as a single pill or as two disease or condition being treated, and the judgment of the separate pills). One of the therapeutic agents may be given in practitioner. multiple doses, or both may be given as multiple doses. If not 0.178 Toxicity and therapeutic efficacy of such therapeutic simultaneous, the timing between the multiple doses may regimens can be determined by Standard pharmaceutical pro vary from more than Zero weeks to less than four weeks. In cedures in cell cultures or experimental animals, including, addition, the combination methods, compositions and formu but not limited to, the determination of the LD50 (the dose lations are not to be limited to the use of only two agents; the lethal to 50% of the population) and the ED50 (the dose use of multiple therapeutic combinations is also envisioned. therapeutically effective in 50% of the population). The dose 0183 The pharmaceutical agents which make up the com ratio between the toxic and therapeutic effects is the thera bination therapy disclosed herein may be a combined dosage peutic index and it can be expressed as the ratio between form or in separate dosage forms intended for Substantially LD50 and ED50. Compounds exhibiting high therapeutic simultaneous administration. The pharmaceutical agents that indices are preferred. The data obtained from cell culture make up the combination therapy may also be administered assays and animal studies can be used in formulating a range sequentially, with either therapeutic compound being admin of dosage for use in human. The dosage of Such compounds istered by a regimen calling for two-step administration. The lies preferably within a range of circulating concentrations two-step administration regimen may call for sequential that include the ED50 with minimal toxicity. The dosage may administration of the active agents or spaced-apart adminis vary within this range depending upon the dosage form tration of the separate active agents. The time period between the multiple administration steps may range from, a few min employed and the route of administration utilized. utes to several hours, depending upon the properties of each pharmaceutical agent, Such as potency, solubility, bioavail Combination Treatments ability, plasma half-life and kinetic profile of the pharmaceu 0179 The neuroprotective compound compositions tical agent. Circadian variation of the target molecule con described herein can also be used in combination with other centration may also determine the optimal dose interval. therapeutic reagents that are selected for their therapeutic 0184. In addition, the compounds described herein also value for the condition to be treated. In general, the compo may be used in combination with procedures that may pro sitions described herein and, in embodiments where combi vide additional or synergistic benefit to the patient. By way of national therapy is employed, other agents do not have to be example only, patients are expected to find therapeutic and/or administered in the same pharmaceutical composition, and prophylactic benefit in the methods described herein, wherein may, because of different physical and chemical characteris pharmaceutical composition of a compound disclosed herein tics, have to be administered by different routes. and/or combinations with other therapeutics are combined 0180. In certain instances, it may be appropriate to admin with genetic testing to determine whether that individual is a ister at least one neuroprotective compound described herein carrier of a mutant gene that is known to be correlated with in combination with another therapeutic agent. By way of certain diseases or conditions. example only, if one of the side effects experienced by a 0185. The compounds described herein and combination patient upon receiving one of the neuroprotective compounds therapies can be administered before, during or after the described herein is nausea, then it may be appropriate to occurrence of a disease or condition, and the timing of admin administer an anti-nausea agent in combination with the ini istering the composition containing a compound can vary. tial therapeutic agent. Or, by way of example only, the thera Thus, for example, the compounds can be used as a prophy peutic effectiveness of one of the compounds described lactic and can be administered continuously to Subjects with herein may be enhanced by administration of an adjuvant a propensity to develop conditions or diseases in order to (i.e., by itself the adjuvant may have minimal therapeutic prevent the occurrence of the disease or condition. A com benefit, but in combination with another therapeutic agent, pound is preferably administered as soon as is practicable the overall therapeutic benefit to the patient is enhanced). Or, after the onset of a disease or condition is detected or Sus by way of example only, the benefit experienced by a patient pected, and for a length of time necessary for the treatment of may be increased by administering one of the compounds the disease, Such as, for example, from about 1 month to about described herein with one or more (e.g., one, two, or three) 3 months. The length of treatment can vary for each subject other therapeutic agents (which also includes a therapeutic For example, the compound or a formulation containing the regimen) that also have a therapeutic benefit. In any case, compound can be administered for at least 2 weeks, about 1 regardless of the disease, disorder or condition being treated, month to about 5 years, and from about 1 month to about 3 the overall benefit experienced by the patient may simply be years. additive of the multiple therapeutic agents or the patient may Exemplary Therapeutic Agents for Use in Combination with experience a synergistic (i.e., a greater than additive) benefit a Neuroprotective Compound due to their specific combination. 0181. The compounds may be administered concurrently Agents for Treating Multiple Sclerosis (e.g., simultaneously, essentially simultaneously or within 0186. Where a subject is suffering from or at risk of suf the same treatment protocol) or sequentially, depending upon fering from multiple Sclerosis, a neuroprotective compound the nature of the disease, disorder, or condition, the condition disclosed herein can be used together with one or more of the of the patient, and the actual choice of compounds used. following exemplary multiple Sclerosis therapeutic agents in US 2010/00566.17 A1 Mar. 4, 2010 22 any combination: Interferon B-1a, Interferon B-1b, glatiramer can be used together with one or more of the following thera acetate (Copaxone(R), mitoxantrone (Novantrone(R), low peutic agents in any combination: immunosuppressants (e.g., dose naltrexone, Nataliaurab (Tysabri R), SativeX(R), Aimspro tacrolimus, cyclosporin, rapamicin, methotrexate, cyclo (Goats Serum), Trimesta (Oral Estriol), Laquinimod, phosphamide, azathioprine, mercaptopurine, mycopheno FTY720 (Fingolimod), MBP8298, NeuroVaxTM, TovaxinTM, late, or FTY720), glucocorticoids (e.g., prednisone, cortisone Revimmune, CHR-1103, BHT-3009, BG-12, Cladribine, acetate, prednisolone, methylprednisolone, dexamethasone, daclizumab (Zenapax) Rituximab (Rituxan), cyclophospha betamethasone, triamcinolone, beclometasone, fludrocorti mide, Campath, Fampridine-SR, MN-166, Temsirolimus, or Sone acetate, deoxycorticosterone acetate, aldosterone), non RPI-78M. steroidal anti-inflammatory drugs (e.g., Salicylates, aryla Agents for Treating Dementia (e.g. Alzheimer's Disease or llknoic acids, 2-arylpropionic acids, N-arylanthranilic acids, AIDS-Related Dementia) oxicams, coxibs, or Sulphonanilides), Cox-2-specific inhibi 0187. Where a subject is suffering from or at risk of suf tors (e.g., Valdecoxib, celecoxib, or rofecoxib), leflunomide, fering from dementia, a neuroprotective compound disclosed gold thioglucose, gold thiomalate, aurofin, Sulfasalazine, herein can be used together with one or more agents or meth hydroxychloroquinine, minocycline, TNF-C. binding pro ods for treating dementia in any combination. Examples of teins (e.g., infliximab, etanercept, oradalimumab), abatacept, therapeutic agents/treatments for treating dementia include, anakinra, interferon-B, interferon-Y, interleukin-2, allergy but are not limited to any of the following FlurizanTM (MPC vaccines, antihistamines, antileukotrienes, beta-agonists, 7869, r flurbiprofen), memantine, galantamine, rivastigmine, theophylline, or anticholinergics. donezipil, tacrine, AB immunotherapy, resveratrol, (-)- epigallocatechin-3-gallate, statin, vitamin C, or vitamin E. Agents for Treating Thromboembolic Disorders 0.192 Where a subject is suffering from or at risk of suf Agents for Treating Parkinson's Disease fering from a thromboembolic disorder (e.g., stroke), the 0188 Where a subject is suffering from or at risk of suf Subject can be treated with a neuroprotective compound dis fering from Parkinson's Disease, a neuroprotective com closed herein in any combination with one or more other pound disclosed hereincan be used together with one or more anti-thromboembolic agents. Examples of anti-thromboem agents or methods for treating Parkinson's disease in any bolic agents include, but are not limited any of the following: combination. Examples of therapeutic agents/treatments for thrombolytic agents (e.g., alteplase anistreplase, streptoki treating Parkinson's Disease include, but are not limited to nase, urolinase, or tissue plasminogen activator), heparin, any of the following: L-dopa, carbidopa, benserazide, tolca tinZaparin, warfarin, dabigatran (e.g., dabigatran etexilate). pone, entacapone, bromocriptine, pergolide, pramipexole, factor Xa inhibitors (e.g., fondaparinux, draparinux, rivar ropinirole, cabergoline, apomorphine, lisuride, selegiline, or oxaban, DX-9065a, otamixaban, LY517717, or YM150), rasagiline. ticlopidme, clopidogrel, CS-747 (prasugrel, LY640315), Ximelagatran, or BIBR 1048. Agents for Treating Amyotrophic Lateral Sclerosis 0189 Where a subject is suffering from or at risk of suf Agents for Treating an HV Infection fering from Amyotrophic Lateral Sclerosis, a neuroprotective (0193 Where the subject is suffering from an EHIV infec compound disclosed herein can be used together with one or tion (e.g., Suffering from AIDS), any of the neuroprotective more agents or methods for treating Amyotrophic Lateral compounds disclosed herein can be administered to the Sub Sclerosis in any combination. Examples of therapeutic ject prophylactically or therapeutically to treat AIDS-related agents/treatments for treating Parkinson's Disease include, dementia in combination with one or more anti-HIV com but are not limited to any of the following: riluzole, insulin pounds administered to treat the HIV infection. Examples of like growth factor 1, or ketogenic diet. anti-HIV compounds include, but are not limited to, AZT (zidovudine, Retrovir), ddI (didanosine, Videx), 3TC (lami Agents for Treating Huntington's Disease Vudine, Epivir), d4T (stavudine, Zerit), abacavir (Ziagen), 0190. Where a subject is suffering from or at risk of suf and FTC (emitricitabine, Emtriva), tenofovir (Viread), fering from Huntington's Disease, a neuroprotective com efavirenz (Sustiva), nevirapine (Viramune), lopinavir/ pound disclosed hereincan be used together with one or more ritonavir (Kaletra), indinavir (Crixivan), ritonavir (Norvir), agents or methods for treating Huntington's Disease in any nelfinavir (Viracept), saquinavir hard gel capsules (Invirase), combination. Examples of therapeutic agents/treatments for atazanavir (ReyataZ), amprenavir (Agenerase), fosam treating Huntington's Disease include, but are not limited to prenavir (Telzir), tipranavir (Aptivus), or T20 (enfuvirtide, any of the following: dopamine receptor blockers, creatine, Fuzeon) CoQ10, minocycline, exercise, antioxidants, antidepressants (notably, but not exclusively, selective serotonin reuptake Antipsychotic Compounds inhibitors SSRIs, such as sertraline, fluoxetine, and paroxet 0194 Where the subject is suffering from schizophrenia, ine), dopamine antagonists, (e.g., tetrabenazine), or RNAi which has recently been found to be characterized by a pro mediated silencing of mutant Huntingtin expression. gressive neurodegenerative process (see, e.g., Perez-Neri et Agents for Treating Autoimmune Inflammatory, or Allergic al. (2006), Neurochem Res, 31(10): 1279-1294), any of the conditions neuroprotective compounds disclosed herein can be admin 0191) Where a subject is suffering from or at risk of suf istered to the subject prophylactically or therapeutically in fering from an autoimmune, inflammatory disease, or allergic combination with one or more antipsychotic compounds for condition that affects the nervous system (see, e.g., Allanetal. treatment of schizophrenia. Examples of antipsychotic com (2003), Philos Trans R Soc Lond B Biol Sci, 358(1438): pounds include, but are not limited to, clozapine, risperidone, 1669-1677), a neuroprotective compound disclosed herein olanzapine, quetiapine, Ziprasidone, aripiprazole, paliperi US 2010/00566.17 A1 Mar. 4, 2010 done, sertindole, Zotepine, amisulpride, bifeprunoX, melper tial compounds (I.e., those exhibiting neuroprotective activ one, chlorpromazine (largactil, thorazine), fluphenazine, ity), we selected a further limited group of modified terpe haloperidol, molindone, thiothixene, thioridazine, trifluop noids. A larger series of modified terpenoid compounds was erazine, loxapine, perphenazine, prochlorperazine, then screened in the same type of assay as described below. pimozide, thiothixene, or Zuclopenthixol. Exemplary neuroprotective modified terpenoids are shown in FIG 2. Antiepileptic Compounds 0.195. Where the subject is suffering from epilepsy, any of Example 2 the neuroprotective compounds disclosed herein can be administered to the Subject prophylactically or therapeuti Pharmaceutical Compositions cally in combination with one or more antiepileptic com pounds. Examples of antiepileptic compounds include, but Example 2a are not limited to, carbamazepine, clobazam, clonazepam, ethoSuximide, felbamate, fosphenyloin, flurazepam, gabap Parenteral Composition entin, lamotrigine, levetiracetam, Oxcarbazepine, mepheny 0199 To prepare a parenteral pharmaceutical composition loin, phenobarbital, phenyloin, pregabalin, primidone, suitable for administration by injection, 100 mg of a water Sodium valproate, tiagabine, topiramate, Valproate semiso soluble salt of a compound of any of Formula I, Formula II, dium, Valproic acid, vigabatrin, diazepam, or lorazepamr. Table 2 or Table 3, is dissolved in DMSO and then mixed with 10 mL of 0.9% sterile saline. The mixture is incorporated into Neuroprotective Compounds and Compositions a dosage unit form Suitable for administration by injection. 0196. In some embodiments, one or more of the neuropro tective modified terpenoid compounds disclosed herein can Example 2b be used in combination with one or more neuroprotective compounds or compositions to treat a Subject Suffering from Oral Composition or at risk of neurodegenerative condition. Examples of neu roprotective compounds include, but are not limited to, any of 0200. To prepare a pharmaceutical composition for oral the following: resveratrol, GPI 1046, epigallocatechin gal delivery, 100 mg of a compound of any of Formula I, Formula late, C.-lipoic acid, Omega-3 fatty acids (e.g., docosa II, Table 2 or Table 3, is mixed with 750 mg of starch. The hexaenoic acid or eicosapentaenoic acid), Vitamin E (toco mixture is incorporated into an oral dosage unit for, such as a pherol), carnitine, cytidine diphosphocoline (citicholine), hardgelatin capsule, which is suitable for oral administration. coenzyme Q10, curcumin, Salviolonic acid B, folic acid, Gingko biloba extract, ginsenoside Rb1, ginsenoside Rg3. Example 2c L-Glutathione, grape seed extract, lutein, Zeaxanthin, meth ylcobalamin, N-acetyl-L-cysteine, pycnogenol, quercetin, or Sublingual (Hard Lozenge) Composition taurine. 0201 To prepare a pharmaceutical composition for buccal EXAMPLES delivery, Such as a hard lozenge, mix 100 mg of a compound of any of Formula I, Formula II, Table 2 or Table 3, with 420 0197) The following specific examples are to be construed mg of powdered Sugar mixed, with 1.6 mL of light corn Syrup, as merely illustrative, and not limitative of the remainder of 2.4 mL distilled water, and 0.42 mL mint extract. The mixture the disclosure in any way whatsoever. Without further elabo is gently blended and poured into a mold to form a lozenge ration, it is believed that one skilled in the art can, based on the suitable for buccal administration. description herein, utilize the present invention to its fullest extent. All publications cited herein are hereby incorporated by reference in their entirety. Where reference is made to a Example 2d URL or other such identifier or address, it is understood that Inhalation Composition Such identifiers can change and particular information on the internet can come and go, but equivalent information can be 0202) To prepare a pharmaceutical composition for inha found by searching the internet Reference thereto evidences lation delivery, 20 mg of a compound of any of Formula I, the availability and public dissemination of such information. Formula II, Table 2 or Table 3, is mixed with 50 mg of anhydrous citric acid and 100 mL of 0.9% sodium chloride Example 1 Solution. The mixture is incorporated into an inhalation deliv ery unit, such as a nebulizer, which is suitable for inhalation Identification of Neuroprotective Compounds administration. 0198 We sought to identify neuroprotective compounds using an in vitro neuroprotection assay as described in detail Example 2e below. We screened a specific collection/library of com pounds that we identified as potentially active agents. This Rectal Gel Composition particular collection contains 2000 compounds of which 50% are FDA-approved compounds, 30% are natural products, 0203 To prepare a pharmaceutical composition for rectal and 20% are other bioactive compounds. As shown in FIG. 1, delivery, 100 mg of a compound of any of Formula I, Formula the vast majority of the compounds conferred little or no II, Table 2 or Table 3, is mixed with 2.5g of methylcelluose neuroprotection or were even neurotoxic, with only about 3% (1500 mPa), 100 mg of methylparapen, 5 g of glycerin and exhibiting neuroprotective activity. From this subset of poten 100 mL of purified water. The resulting gel mixture is then US 2010/00566.17 A1 Mar. 4, 2010 24 incorporated into rectal delivery units, such as Syringes, 0208 Mixed hippocampal cultures were incubated with which are suitable for rectal administration. 3-NP (0.5-10 mM) for 18 hours and then assessed for viability using an MTT assay. As shown in FIG. 3, titration of 3-NP Example 2f levels for neurotoxic effects demonstrated that 3 mM 3-NP treatment consistently induced 25-35% cytotoxicity in rat Topical Gel Composition mixed hippocampal cultures. 0209. The assay system was validated using two neuro 0204 To prepare a pharmaceutical topical gel composi protective agents, GPI 1046 and Resveratrol. Both of these tion, 100 mg of a compound of any of Formula I, Formula II, compounds has demonstrated antioxidant and/or neuropro Table 2 or Table 3, is mixed with 1.75 g of hydroxypropyl tective activities in numerous in vitro and in vivo assays (for cellulose, 10 mL of propylene glycol, 10 mL of isopropyl review, see Poulter et al. (2004), Neuroscience, 128(1):1-6: myristate and 100 mL of purified alcohol USP. The resulting Caporello, etal. (2006), J Neurochem, 98(1): 146-155; Zamin gel mixture is then incorporated into containers, such as et al. (2006), Neurobiol Dis, 24(1):176-182). Cultures were tubes, which are suitable for topical administration. preincubated with GPI 1046 or Resveratrol for one hour prior to an 18 hour exposure to 3 mM 3-NP. These “positive con Example 2g trol’ neuroprotective compounds significantly protected rat Ophthalmic Solution Composition neurons from oxidative damage elicited by 3-NP (FIG. 4A) in the rat mixed hippocampal culture assay system described 0205 To prepare a pharmaceutical opthalmic solution above. The same neuroprotective compounds were evaluated composition, 100 mg of a compound of any of Formula I, for efficacy against HIV-1 Tat protein toxicity using the same Formula II, Table 2 or Table 3, is mixed with 0.9 g of NaCl in 1 hour preincubation protocol. As with the 3-NP neurotoxic 100 mL of purified water and filtered using a 0.2 micron filter. ity assay, these compounds protected hippocampal neurons The resulting isotonic solution is then incorporated into oph from Tattoxicity as well (FIG. 4B). These data indicated that thalmic delivery units, such as eye drop containers, which are the measurement of neuroprotection against 3-NP toxicity suitable for ophthalmic administration. Example 3: Biologi likely serves as a good indicator of protective activity against cal Assays and Analyses HIV-1 Tattoxicity. 0210. Using the validated 3 mM 3-NP neurotoxicity assay Example 3A described above, we evaluated the neuroprotective efficacy of Exemplary Modified Terpenoids are Neuroprotective approximately 2000 compounds from the Spectrum Collec against an Oxidative Stressor and a Neurotoxic Pro tion (MicroSource Discovery) as described in Example 1. tein Several of the neuroprotective compounds identified in this collection were modified terpenoids. Thus, we tested an 0206 We sought to evaluate the protective efficacy of a expanded collection of modified terpenoids in the in vitro number of modified terpenoid compounds against the against neuroprotection assay. As shown in Tables 2 and 3, a number many different neurotoxins, ranging from the chemotoxic of modified terpenoid compounds were identified as having 6-OHDA, NMDA, 3-nitropropionic acid (3-NP), and viral neuroprotective activity against 3-NP. proteins such as Tat and gp120. Thus, we established an in 0211 3-NP assay data for four of these compounds are vitro neuroprotection assay using rat mixed hippocampal cul shown in FIG. 5. These compounds protected the mixed hip tures, in which we evaluated the protective efficacy of neuro pocampal cultures significantly, with nearly complete protec protective compounds disclosed herein. The oxidative stres tion provided by 10LM of Khivorin, about 60-70% protection sor 3-NP was used to elicit toxicity in the rat hippocampal resulting from odoratone, and about 50% protection from cultures to mimic the oxidative damage, reactive oxygen spe gedunin and angolensic acid, methyl ester treatment. The cies production and ensuing neurodegeneration resulting 3-NP protection dose-response characteristics of the modi from HIV infection. Another measure of neurotoxicity which fied terpenoids Khivorin, Gedunin, Nomin, and Limonin are results from HIV infection was evaluated by exposure of the shown in FIGS. 6A-6D, respectively. hippocampal cultures to HIV-1 Tat (Lietal (2005), Neurotox 0212 Gedunin and Limonin also dose dependently pro Res, 8(1-2): 119-134). tected hippocampal cultures from HIV-1 Tattoxicity (FIG. 7), 0207 Rat mixed hippocamal neuronal cultures were gen with nearly complete neuroprotection provided by 1-10 uM erated from freshly dissected rathippocampi (embryonic day Gedunin. Thus, Some modified terpenoids also protect hip 18) in neurobasal media containing 5% fetal bovine serum pocampal neurons from HIV-1 neurotoxic protein degenera and 2% B27 supplement. The cells were plated into 96 well tion. plates at a density of 4x10 cells/mL and routinely used on 0213 We also tested the ability of Limonin to protect days 11-14 following culturing. Cell viability was assessed against N-methyl-D-Aspartic Acid (NMDA) excitotoxicity, with MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetra and 6-Hydroxydopamine (6-OHDA), a dopaminergic neuro Zolium bromide assay. The MTT assay is based on the ability toxin. As shown in FIG. 8, Limonin dose-dependently of a mitochondrial dehydrogenase enzyme from viable cells blocked the neurotoxicity of both NMDA (FIG. 8A) and to cleave the tetrazolium rings of the pale yellow MTT and 6-OHDA (FIG. 8B) conferring complete protection at the form dark blue formazan crystals. See Mosmann (1983), J highest Limonin dose tested (10 uM). Immunol Methods, 65(1-2):55-63. These crystals are largely 0214 Finally, we sought to determine if the compounds impermeable to cell membranes, and thus accumulate within tested in rat neuronal cultures would also be effective on healthy cells. The resultant formazan precipitates are solubi cultured human fetal neurons. Indeed, as shown in FIG. 9, lized with DMSO and read on a multiwell scanning spectro Limonin dose-dependently protected human fetal neuronal photometer (ELISA reader). The number of surviving cells is cultures against both 3-NP (FIG.9A) and 6-OHDA. directly proportional to the level of the formazan product 0215 Based on these data, we concluded that various created. modified terpenoid compounds are neuroprotective. US 2010/00566.17 A1 Mar. 4, 2010 25
Compound Structure % Protection
3alpha 151 ACETOXYDIHYDRODEOXY. GEDUNIN
1,3-DIDEACETYLKHIVORIN 129
DEOXODIHYDROGEDUNIN 101
3beta 128 ACETOXYDEOXYANGOLENSIC ACID, METHYL ESTER US 2010/00566.17 A1 Mar. 4, 2010 26
-continued
Compound Structure % Protection
TRIDESACETOXYKHIVORIN 97
7beta-HYDROXY-7- O O 84 DESACETOXYKHIVORINIC N-1N
ACID, METHYL ESTER 1. E
3beta-HYDROXYDEOXODIHYDRO- W O 82 GEDUNIN 2
GEDUNINDEOXODEOXYDIHYDRO- CO 77
US 2010/00566.17 A1 Mar. 4, 2010 27
-continued
Compound Structure % Protection
KHIVORIN 70
EPOXYGEDUNIN 69
7-EPIKHIVORIN 65
3beta,7beta 61 DIACETOXYDEOXODEACETOXY. DEOXYDIHYDROGEDUNIN
US 2010/00566.17 A1 Mar. 4, 2010 28
-continued
Compound Structure % Protection
DESACETYL (7)KHIVORINIC 58 ACID, METHYL ESTER
3-DEOXO-3beta 56 ACETOXYDEOXYDIHYDRO GEDUNIN
3beta 56 HYDROXYDEOXODIHYDRO DEOXYGEDUNIN
DEOXYGEDUNOLACETATE
US 2010/00566.17 A1 Mar. 4, 2010 29
-continued
Compound Structure % Protection
ISOGEDUNIN 50
GEDUNOL 45
2,3-DIHYDROISOGEDUNIN 44
7-DEACETOXY7-OXO 42 KHIVORINIC ACID, METHYL ESTER
US 2010/00566.17 A1 Mar. 4, 2010 30
-continued
Compound Structure % Protection
TRIDESACETOXYKHIVORIN 39
3beta-ACETOXYDEOXODIHYDRO- W O 39 GEDUNIN 21
DEACETOXY-7-OXOGEDUNIN C 38
DEOXYKHIVORIN 36 US 2010/00566.17 A1 Mar. 4, 2010 31
-continued
Compound Structure % Protection
7-DEACETOXY-7- 36 OXOKHIVORIN
3alpha-HYDROXY-3- 35 DEOXYANGOLENSIC ACID METHYLESTER
ANGOLENSIC ACID, METHYL O 33
ESTER C)
7-DEACETYLKHIVORIN
US 2010/00566.17 A1 Mar. 4, 2010 32
-continued
Compound Structure % Protection
3beta 30 HYDROXYDEOXYDESACETOXY. 7-OXOGEDUNIN
3-alpha 28 HYDROXYDEOXYGEDININ
DIHYDROGEDUNIN 28
6-HYDROXYANGOLENSIC 26 ACID METHYLESTER
US 2010/00566.17 A1 Mar. 4, 2010 33
-continued
Compound Structure % Protection
12alpha 25 EPOXYDEACETOXYDIHYDRO GEDUNIN
7-DEACETOXY-7- 24 OXODEOXYGEDUNIN
DEOXYGEDUNIN 23
GEDUNIN 40
US 2010/00566.17 A1 Mar. 4, 2010 34
-continued
Compound Structure % Protection
DEACETYLGEDUNIN 19
DIHYDROGEDUNIN 19 ETHANEDITHIOKETAL
1,7-DIDEACETOXY-1,7- 15 DIOXO-3- DEACETLYKHIVORN
1,3-DIDEACETYL-7- 15 DEACETOXY-7- OXOKHIVORIN
US 2010/00566.17 A1 Mar. 4, 2010 35
-continued
Compound Structure % Protection 1 (2)alpha 14 EPOXYDEOXYDIHYDRO GEDUNIN
compartment of semipermeable TranswellTM tissue culture TABLE 3 inserts (24 wells, Corning-Costar). The upper compartment compares to blood side and bottom compartment to brain Modified Terpenoids Exhibiting Neuroprotective Activity against 3-NP side. Transmission electron microscopy revealed a smooth endothelial cell monolayer, typical rod shaped Weibel-Palade Protection bodies and tightjunctions. Polarity was shown after treatment Structure vs 3-NP with TNF-C., which resulted in an apical expression of ICAM 3beta-ACETOXYDEOXODIHYDROGEDUNIN 39 1, which is in agreement with data of Wong et al. (1992), J CARAPIN-8(9)-ENE 38 Neuroimmunol, 39(1-2):11-21. The presence of junctional DEACETOXY-7-OXOGEDUNIN 38 proteins is seen by ZO1 immuno-staining and Western blot KHAYANTHONE 36 DEOXYKHIVORIN 36 ting for ZO-1, beta-catenin and occludin-1, showing that 7-DEACETOXY7-OXOKHIVORIN 36 HBMEC possess endothelial and brain characteristics and ANGOLENSIC ACID, METHYL ESTER 33 functions. 7-DEACETYLKHIVORIN 32 0217 Propidium iodide (PI) (MW=600) at 0.5 mg/ml, an FISSINOLIDE 28 6-HYDROXYANGOLENSIC ACID METHYLESTER 26 indicator of in vitro blood brain barrier (“BBB) integrity, is DEOXYGEDUNIN 23 applied to the upper compartment along with a vehicle solu DEACETYLGEDUNIN 19 tion or a solution containing test compound at a concentration BUSSEIN 18 of 1 M. At 2 and 4 hours post drug treatment, levels of CAR.APIN 18 compound are measured in top and bottom compartments by ENTANDROPHRAGMIN 18 UTILIN 16 mass spectrometery as described in Tian et al. (2004), Rapid 3-DIDEACETYL-7-DEACETOXY-7-OXOKHIVORIN 15 Comm Mass Spec, 18:3099-3104. The level of test compound 7-DIDEACETOXY-1,7-DIOXO-3-DEACETYLKHIVORIN 15 detected in the bottom compartment is then normalized for KHAYASINC 13 differences in PI permeability between the test compound DIHYDROFISSINOLIDE 12 solution and the vehicle control solution. 8beta-HYDROXYCARAPIN, 3,8-HEMIACETAL 9 MEXICANOLIDE 9 QUASSIN 8 Example 3C 3-DEACETYLKHIVORIN 7 PRIEURIANIN 6 In Vivo Assessment of Neuroprotective Modified Terpenoid Compounds in a 3-NP-Induced Neurotox icity Animal Model Example 3B 0218. In order to determine the in vivo efficacy of com pounds identified as neuroprotective against 3-NP in vitro, as In Vitro Modeling of Blood Brain Barrier Permeabil described herein, we employ a 3-NP-induced neurotoxicity ity to Test Compounds model in rats. See, e.g., Kumar et al. (2006), Behav Pharma 0216. In vitro models of the Blood Brain Barrier from col, 17(5-6):485-492. human brain microvascular endothelial cells (HBMEC) that 0219. To induce 3-NP neurotoxicity in vivo, 12-week-old were isolated and characterized have been published previ male Lewis rats weighing 340-370gm are administered intra ously (see, e.g., Stins et al. (1997), J Neuroimmunol, 76(1-2): peritoneal injections of 3-NP (20 mg/kg) for 4 days. The 81-90; Cucullo et al. (2007), Epilepsia, 48(3):505-516). animals are divided into three treatment groups (n=12 per These HBMEC possess gamma glutamyl transpeptidase group) as follows: (GGTP) and drug transporter P-glycoprotein, and junctional 0220 Group 1 is administered Khivorin, Gedunin, Odo proteins as seen by ZO1 immuno-staining, thereby demon ratone, Angolensic acid, or another modified terpenoid com strating their brain endothelial cell characteristics. In vitro pound once daily (10 mg/kg, oral gavage) beginning four BBB models were constructed by growing HBMEC on days prior to and continuing for four days Subsequent to the microporous membranes (0.4 um pore size) in the upper beginning of the 3-NP injections. US 2010/00566.17 A1 Mar. 4, 2010 36
0221 Group 2 is administered saline vehicle (negative with TNF-C. and Y-IFN to induce a focal EAE lesion as control), by oral gavage, beginning four days prior to and described below. Three weeks later, the animals are sacri continuing for four days Subsequent to the beginning of the ficed. Outcome measures include behavioral studies (Basso 3-NP injections. Beattie-Bresnahan scale), evoked potentials (nerve conduc 0222 Group 3 is administered resveratrol (positive con trol) (10 mg/kg, oral gavage) by oral gavage, beginning four tion Velocity), radiological outcomes (Diffusion Tensor days prior to and continuing for four days Subsequent to the Imaging, Magnetic Resonance Imaging), and histology mea beginning of the 3-NP injections. surements (Luxol Fast Blue, Toluidine Blue, Myelin Basic 0223 Subsequent to the beginning of the 3-NP injections, Protein, phosphoNeurofilament and AXonal degeneration). A animals are assessed for significant loss of body weight, a second cohort includes another set of animals where the same decline in motor function (locomotor activity, movement pat drug regimen listed above is initiated following the laminec tern, and vacuous chewing movements) and cognitive deficits tomy. (e.g., impairment in learning or memory). Differences of performance between groups are analyzed at each time point Animals by two-tailed ttest 0224. Twenty-four hours after the last 3-NP injection ani 0229. For these experiments, Lewis rats (10-12 weeks old) mals are anesthetized, perfused transcardially with Saline, are allowed free access to food and water to acclimate 7-10 followed by ice-cold 4% paraformaldehyde in 0.1 M phos days before the initiation of experiments. At the time of the phate buffer, pH 7.4. Brains are immediately removed and study, the animals weigh 200-250 g. postfixed overnight in the same fixative and then cryopro tected in 30% sucrose in 0.1 M phosphate buffer, pH 7.4. Induction of Myelin Oligodendrocyte Glycoprotein (MOG) Sequential coronal sections (30 um) are made on a freezing Sensitivity microtome, starting from the anterior aspect of the corpus callosum throughout the entire striatum. For histological 0230 Animals are injected subcutaneously at the base of assessment, every sixth section (210um interval) is processed the tail with 100 ul of recombinant MOGs (250-500 for cresyl Violet staining to assess cell loss and neuronal ug/mL) emulsified in incomplete Freund's adjuvant. No degeneration. Cresyl Violet staining is performed with stan manipulations are performed for 18-30 days after immuniza dard protocols. tion to allow for the immune system to develop a sensitivity to 0225 Stereological analysis of lesion volumes are per MOG. This procedure does not induce clinical symptoms of formed by digitally acquiring cresyl violet-stained sections EAE. through the striatum at 4x objective using a computerized image analysis system. Lesion Volumes for each group of Induction of Focal EAE Lesion animals are calculated by Summing the cross-sectional areas of the lesion in each section and multiplying this value by the 0231. To induce an EAE lesion within a specific region of distance between sections. the spinal cord, a steriotaxic injection of TNF-C. and IFN-Y is 0226 Compounds found to confer a significant reduction administered within the spinal cord after laminectomy. The in 3-NP-induced behavioral deficits or neuroanatomical animals are shaved prior to initiation of the procedure. Anes lesion volume are considered to be neuroprotective in vivo. thetized animals are placed in the prone position, and a mid line incision is made. Following the dissection of fascia and Example 3D muscle, a laminectomy is performed at T8 with Rongeur In Vivo Assessment of Identified Neuroprotective forceps. A focal EAE lesion within the cortical spinal tract is Compounds in a Focal Ischemia Animal Model induced by administering 2 LL of 250 ng of TNF-C. and 150 Units of IFN-y dissolved in phosphate buffered saline with 0227. A problem with ongoing methodology used to test trace amounts of Monastral Blue using a capillary glass tube. the efficacy of neuroprotective compounds in vivo in the Following surgery the wound is sutured first through the classic Experimental Autoimmune Encephalomyelitis (EAE) model is that inflammation-mediated lesions are randomly fascia, and then the skin with a 4.0 Vicryl thread or with distributed within the central nervous system, thereby making wound clips. Animals are warmed and allowed to recover. uniform quantification of axonal damage and neuronal cell 0232. It is understood that the examples and embodiments death difficult. In contrast, a focal lesion model induces a described herein are for illustrative purposes only and that localized inflammatory response and Subsequent lesion only various modifications or changes in light thereofwill be Sug within a specific region of the spinal cord. The focal lesion gested to persons skilled in the art and are to be included model therefore enables a more accurate assessment of the within the spirit and purview of this application and scope of efficacy of neuroprotective agents because a specific region of the appended claims. All publications, patents, and patent the spinal cord and brain are compared between animals. applications cited herein are hereby incorporated by refer Furthermore, the focal lesion model decreases variability ence in their entirety for all purposes. found within the classic EAE models and therefore decreases the number of animals required. 0228 Lewis rats (12-15 rats per treatment group) are treated with a test compound at a dose of 0.1. 1, and 10 mg/kg 1-11. (canceled) s.c or p.o. or with vehicle once daily for 3 days prior to 12. A method for treating or reducing the risk of a neuro immunization with MOG and incomplete Freunds Adjuvant degenerative condition in a subject in need thereof, compris as described below. Eighteen days later, the animals are sub ing administering to the Subject a therapeutically effective jected to laminectomy of the dorsal column at T8 and injected amount of the pharmaceutical composition comprising US 2010/00566.17 A1 Mar. 4, 2010 37
23. The method of claim 22, wherein the acute neurode
generative condition is stroke. 24. The method of claim 23, wherein the stroke is selected from an acute thromboembolic stroke, a focal ischemia, a global ischemia, or a transient ischemic attack. 25. The method of claim 22, wherein the acute neurode generative condition is an ischemia resulting from a Surgical technique involving prolonged halt of blood flow to the brain. 26. The method of claim 22, wherein the acute neurode generative condition is selected from head trauma, spinal trauma, optic nerve stroke, anterior ischemic optic neuropa thy, or traumatic optic neuropathy. 27. The method of claim 16, wherein the chronic neurode generative condition is glaucoma, optic neuritis, compressive optic neuropathy, or a hereditary neuropathy. 28. The method of claim 16, wherein the chronic neurode generative condition is schizophrenia. 13. The method of claim 12, wherein, at a concentration of 29. A method for treating multiple sclerosis in a subject in 10 uM, the compound provides at least about 20% protection need thereof, comprising administering to the Subject a com against 3 mM3-nitropropionic acid to rat mixed hippocampal position comprising a therapeutically effective amount of the cultures. compound of claim 1. 14. The method of claim 12, wherein the subject in need 30. A method for treating AIDS-related dementia in a sub thereof is diagnosed as Suffering from the neurodegenerative ject in need thereof, comprising administering to the Subject condition prior to the administration. a composition comprising a therapeutically effective amount 15. The method of claim 12, wherein the administration is of the compound comprising parenteral, intravenous, Subcutaneous, intramuscular, trans
nasal, intra-arterial, transdermal, or respiratory. 16. The method of claim 12, wherein the neurodegenera tive condition is a chronic neurodegenerative condition. 17. The method of claim 16, wherein the chronic neurode generative condition is selected from Alzheimer's Disease, multiple sclerosis, Huntington's Disease, or Parkinson's Dis CaSC. 18. The method of claim 16, wherein the chronic neurode generative condition is AIDS related dementia. 19. The method of claim 16, wherein the chronic neurode generative condition is Amyotrophic Lateral Sclerosis. 20. The method of claim 16, wherein the chronic neurode generative condition is a retinal disease. 21. The method of claim 16, wherein the chronic neurode generative condition is epilepsy. 22. The method of claim 12, wherein the neurodegenera tive condition is an acute neurodegenerative condition.