Methods and Compositions for Reducing Neurodegeneration in Amyotrophic Lateral Sclerosis - DocsLib

sign in sign up

<<

Home , Kyotorphin

(19) &

(11) EP 2 255 812 A1

(12) EUROPEAN PATENT APPLICATION

(43) Date of publication: (51) Int Cl.: 01.12.2010 Bulletin 2010/48 A61K 31/575 (2006.01) A61K 47/36 (2006.01) A61K 9/00 (2006.01) A61P 25/16 (2006.01) (2006.01) (21) Application number: 10176111.2 A61P 25/28

(22) Date of filing: 31.10.2005

(84) Designated Contracting States: (72) Inventor: Yoo, Seo Hong AT BE BG CH CY CZ DE DK EE ES FI FR GB GR Wyckoff, HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI New Jersey 07481 (US) SK TR (74) Representative: Lucas, Brian Ronald (30) Priority: 01.11.2004 US 624100 P Lucas & Co. 16.11.2004 US 628421 P 135 Westhall Road Warlingham, (62) Document number(s) of the earlier application(s) in Surrey CR6 9HJ (GB) accordance with Art. 76 EPC: 05820886.9 / 1 814 558 Remarks: This application was filed on 10-09-2010 as a (71) Applicant: Yoo, Seo Hong divisional application to the application mentioned Wyckoff, under INID code 62. New Jersey 07481 (US)

(54) Methods and compositions for reducing neurodegeneration in amyotrophic lateral sclerosis

(57) A composition comprising a clear aqueous so- (c) water, wherein the first material and the carbohydrate lution comprising: both remain in solution for any subset of the range of pH (a) a first material selected from soluble ursodeoxycholic values obtainable in an aqueous system sufficient for a acid or a sodium salt oftauroursodeoxycholic acid (TUD- pharmaceutical formulation to remain in solution from CA); preparation, to administration, to absorption in the body; (b) a carbohydrate selected from the group consisting of an aqueous soluble starch conversion product or an for use in the treatment of at least one symptom of a aqueous soluble non-starch polysaccharide; neurodegenerative disease or at least one symptom of a motor neuron disease in a subject. EP 2 255 812 A1

Printed by Jouve, 75001 PARIS (FR) (Cont. next page) EP 2 255 812 A1

2 1 EP 2 255 812 A1 2

Description closure with riluzole may reduce riluzole toxicity or side effects in some embodiments. TECHNICAL FIELD [0006] Compositions of the present disclosure may include (1) a bile acid, a bile acid derivative, a bile acid salt, [0001] The present disclosure is related to composi- 5 or a bile acid conjugate with an amine, (2) water, and (3) tions and methods for ameliorating or treating at least a sufficient quantity of an aqueous soluble starch con- one symptom of a neurodegenerative process or dis- version product such that the bile acid and the starch ease. conversion product remain in solution at any pH within a selected pH range. BACKGROUND 10 [0007] The disclosure also relates to a composition which comprises (1) a bile acid, a bile acid derivative, a [0002] At any given time, as many as 30,000 Ameri- bile acid salt, or a bile acid conjugate with an amine, (2) cans suffer with Amyotrophic Lateral Sclerosis (ALS), water, and (3) a sufficient quantity of an aqueous soluble which is nearly always fatal. ALS, also known as Lou non-starch polysaccharide such that the bile acid and the Gehrig’s Disease, is a progressive neurodegenerative 15 polysaccharide remain in solution at any pH within a se- disease that attacks motor neurons in the brain and spinal lected pH range. cord and results in muscle weakness and atrophy. Early [0008] The disclosure further relates to a pharmaceu- symptoms include loss of dexterity and gait. As the dis- tical composition which comprises (1) a bile acid, a bile ease progresses, patients becomeparalyzed and require acid derivative, a bile acid salt, or a bile acid conjugate respiratory support. The life expectancy of ALS patients 20 with an amine, (2) water, (3) a pharmaceutical compound is usually 3 to 5 years after diagnosis with the leading in a pharmaceutically appropriate amount, and (4) a suf- cause of death being loss of respiratory function. ficient quantity of an aqueous soluble starch conversion [0003] ALS etiology is only partially understood. Fa- product or an aqueous soluble non-starch polysaccha- milial (inherited) cases make up only about 5-10% of ALS ride such that the bile acid, the pharmaceutical com- patients overall. Within this subset of ALS patients, one 25 pound, and the carbohydrate remain in solution at any in five carry the only genetic defect identified to date, a pH level within a selected pH range. According to some mutation in the SOD1 gene. The mutant allele leads to non-limiting embodiments, the pharmaceutical com- production of a protein believed to be toxic to motor neu- pound may be any drug that has beneficial effect when rons. Most cases, i.e., the remaining 90-95%, arise administered to a subject having a neurodegenerative seemingly spontaneously and without an identifiable pat- 30 disease. According to one non-limiting embodiment of tern. Thus, ALS appears to be capable of striking anyone the disclosure, the pharmaceutical compound may be at any time. Effective therapies are scarce or non-exist- riluzole or pharmaceutically active or activatable metab- ent. olites, pro-drugs, derivatives or analogs of riluzole. [0009] The disclosure further relates to solution dos- SUMMARY 35 age forms of bile acid compositions. Advantages of these solution dosage forms include improved bioavailability [0004] Accordingly, a need has arisen for methods and and absorbability of a bile acid. Additional advantages of compositions useful for ameliorating or eliminating pro- solution dosage forms include improved bioavailability gression of a neurodegenerative process or disease in- and absorbability of a pharmaceutical compound. cluding, without limitation, ALS. 40 [0010] In some embodiments of the disclosure, a com- [0005] The present disclosure relates to compositions position is provided which comprises (1) a bile acid, a and methods for ameliorating or treating at least one bileacid derivative, a bile acid salt, or a bile acid conjugate symptom of a neurodegenerative process or disease. For with an amine, (2) water, and (3) a sufficient quantity of example, in some embodiments, the disclosure provides carbohydrate such that the bile acid component and the compositions and methods for ameliorating or eliminat- 45 carbohydrate remain in solution at any pH within a se- ing progression of a neurodegenerative process or dis- lected pH range, wherein the carbohydrate is a combi- ease. In some embodiments of the disclosure, a clear nation of an aqueous soluble starch conversion product stable solution of a bile acid may be administered to a and an aqueous soluble non-starch polysaccharide. In subject having a progressive neurodegenerative disor- embodiments containing both soluble starch non- der. According to some embodiments, a bile acid solution 50 polysaccharide and high molecular weight starch con- may further comprise another pharmaceutical ( e.g., rilu- version product, the amounts of each are such that when zole). In some embodiments, a bile acid solution of the combined together in the composition they are sufficient disclosure may be administered to a subject having to allow the bile acid component, the high molecular amyotrophic lateral sclerosis. According to some embod- weight starch conversion product, the soluble non- starch iments, coadministration of a bile acid with riluzole may 55 polysaccharide and the pharmaceutical compound, if result in a surprisingly-improved outcome over adminis- any, to remain in solution at any pH within a selected pH tration of either pharmaceutical alone. In some embodi- range. ments, coadministration of a bile composition of the dis- [0011] In some embodiments of the disclosure, a com-

3 3 EP 2 255 812 A1 4 bination therapy composition is provided which may in- been identified as familial ALS (FALS). In about a fifth of crease the intensity of response to or efficacy of a phar- FALS cases, the mutant gene is a cytoplasmic enzyme, maceutical. More specifically, administration of a com- Cu/Zn superoxide dismutase- 1 (SOD). More than 90 mu- position of the disclosure comprising a bile acid and rilu- tations inCu/Zn SOD have been identified andare spread zole to a subject suffering from a neurodegenerative dis- 5 across over 30 sites. These mutations may give rise to order may have more than an additive effect of adminis- a new adverse function that leads to FALS as opposed tration of either compound alone. to simply impairing a normal function of the gene product. For example, in experiments with transgenic mice that BRIEF DESCRIPTION OF THE DRAWINGS overexpress a mutant human Cu/Zn SOD (A4V, G93A, 10 G85R, G37R), there was no correlation between loss of [0012] Some specific example embodiments of the SOD1 activity and the onset or severity of disease. disclosure may be understood by referring, in part, to the [0014] The symptoms and pathology of FALS patients following description and the accompanying drawings, with SOD1 mutations closely resemble those of patients wherein: with SALS. The clinical progression and pathologic alter- 15 ations in motor neurons from mice expressing mutant FIGURE 1A is life expectancy and its result is shown SOD1 are also strikingly similar to those found in SALS as the percent of survival on time when animal died; patients, suggesting that the mechanisms of neurode- FIGURE 1B is Rotarod test and its result is shown generation for SALS and FALS may share common com- as the time they remained on the rod before sliding ponents. Mitochondria play a pivotal role in many meta- off on every week until dying; 20 bolic and apoptotic pathways that regulate the life and FIGURE 2 is a bar graph showing the results of a death of cells. Mitochondria also are the site of initiation cell viability assay with wildtype cells, A4V cells, and of the intrinsic apoptotic cascade, which can be activated G93A cells in which the cells were untreated (left by the release of pro- apoptotic factors that may act both panel) or incubated with 200 nM of solubilized UDCA in a caspase-dependent or caspase-independent man- in solution of the disclosure (center panel), or 20 PM 25 ner. Mitochondrial dysfunction may be directly involved of solubilized UDCA in solution of the disclosure in the pathogenesis of ALS. Mitochondrial dysfunction (right panel); causes motor neuron death by predisposing them to cal- FIGURE 3 is a bar graph showing the results of a cium-mediated excitotoxicity, by increasing generation cell viability assay with wildtype cells, A4V cells, and of reactive oxygen species, and/or by initiating the intrin- G93A cells in which the cells were untreated (left 30 sic apoptotic pathway. panel) or incubated with 500 PM S-nitrosoglutath- [0015] Mitochondrial dysfunction may result in quantal ione (GSNO; middle panel), or 500P M GSNO fol- releases of pro-apoptotic factors, such as cytochrome c, lowed by a 20 PM UDCA solution of the disclosure. apoptosis inducing factor (AIF), and endoG, from individ- FIGURE 4A is a micrograph showing untreated A4V ual mitochondria, perhaps in response to local calcium cells (control cells); 35 mediated toxicity, for example, under excitatory synaps- FIGURE 4B is a micrograph showing A4V cells in- es. This local toxicity might induce death of subcellular cubated with 500 PM S-nitrosoglutathione (GSNO); compartments, e.g., dendritic or axonal branches. This FIGURE 4C is a micrograph showing A4V cells in- kind of subcellular compartmental degeneration might be cubated with 500 PM GSNO and then, were incu- insufficient to induce the cell to die immediately, but could bated in succession with 20 PM of solubilized UDCA 40 spread to the cell bodies over a period of time. Consistent in solution of the disclosure; with this scenario, in ALS, axons degenerate from the FIGURE 4D is a micrograph showing untreated distal to the proximal direction (dying back) and dendrites G93A cells; become atrophic before the final motor neuron death. FIGURE 4E is a micrograph showing G93A cells in- This mechanism of cell death may be unique for neuronal cubated with 500 PM GSNO; and 45 degeneration because neurons have complex subcellu- FIGURE 4F is a micrograph showing G93A cells in- lar branches, and it may progress relatively slowly com- cubated with 500 PM GSNO and then, were incu- pared with typical cell death mechanisms in other cell bated in succession with 20 PM of solubilized UDCA types. As a consequence, it may be that at any given in solution of the disclosure. time in the course of the disease, only a small number 50 of cells are actually dying of apoptosis. DETAILED DESCRIPTION [0016] The motor neuron degeneration by SOD1 mutation may be investigated in cell culture and in transgenic [0013] About 85-90% of adult-onset ALS patients have mice models. In in vitro analyses, the spread and pro- no family history of the disease. This apparently random gressive motor neuronal death may be observed in mis- or haphazard occurrence has lead some practitioners to 55 sense mutations such as G93A (glycine to alanine at po- identify these cases as sporadic ALS (SALS). By con- sition 93) and A4V (alanine to valine at position 4) in the trast, ALS may be inherited as an autosomal dominant human Cu/Zn-superoxide dismutase gene (hSOD1). For condition in about 10-15% of patients. These cases have example, viability of cells with wildtype, G93A, and A4V

4 5 EP 2 255 812 A1 6 hSOD1 24 h after the neuronal differentiation was eval- tageous features of existing commercial dosage forms uated by using both the MTT assay and Trypan blue of UDCA. In addition, a bile acid composition of the dis- staining. Viability was significantly reduced over time. In closure may, in some embodiments, ameliorate and/or the wild-type cells, the viability significantly decreased at treat at least one symptom of ALS and/or advanced ALS. 48 h after the neuronal differentiation (85.91 9.08%) 5 Bile acid dosage forms, according to some embodiments (P < 0.05), and was 59.41 13.54% at 72 h (P < 0.01). of the disclosure, may be suitable or adaptable for oral Viability decreased even more in G93A (63.71 6.25%) and parenteral administration. In some embodiments, a and A4V cells (58.85 7.83%) compared with wild cells bile acid composition of the disclosure may include an (100 6.97%) at 24 h after the neuronal differentiation. intact molecule of UDCA and an aqueous soluble starch At 48 h, viability was further reduced to 23.12 8.96% 10 conversion product ( e.g., a product resulting from hydrol- in G93A cells and 20.79 8.07% in A4V cells (P < 0.01). ysis of starch).A bile acid composition, according to some At 72 h, these mutant cells were nearly all dead with embodiments of the disclosure, may be solubilized in wa- viabilities at about 0%. These results suggest that G93A ter and may remain in aqueous solution without precipi- or A4V mutations in hSOD1 make motor neurons more tation at any pH. vulnerable. In view of the these results, analyses per- 15 [0022] In some embodiments, the solubility of UDCA formed according to the instant disclosure may, in some in a solution of the disclosure may be about 3,000 times embodiments, be performed at about 24 h to avoid the higher than that of commercialized UDCA (0.15mol vs. substantially lower viability that may occur at later time 0.05mmol) and may be 300 times higher than that of points. TUDCA. A solution of the disclosure may, in some em- [0017] The transgenic mice expressing G93A or A4V 20 bodiments, deliver solubilized UDCA to blood, brain, develop a severe motor neuron degenerative syndrome stomach, duodenum, jejunum, ileum and/or colon. In despite normal or above normal SOD activities. By con- some embodiments, an oral and parenteral dosage form trast, these symptoms may not occur in mice in which may contain, for example, 500mg of UDCA and may have Cu/Zn SOD is knocked out or overexpressed. a Cmax that is at least 8 times higher than an existing [0018] Nitric oxide (NO) may play a role in physiolog- 25 commercial UDCA form and Tmax that is about 4-6 times ical and pathological processes in the central nervous shorter than an existing commercial UDCA form. system and mitochondria may be the primary targets for [0023] Moreover, according to some embodiments, a NO toxicity in the brain. NO may interact with a superox- bile composition of the disclosure may not contain any ide anion to form reactive peroxynitrites, which may precipitation at any pH and may function as a systemic cause cell death by oxidative damage, disruption of en- 30 drug. A solution, according to some embodiments, may ergy metabolism, calcium homeostasis, and mitochon- be administered concurrently with one or more pharma- drial function. This toxicity may be prevented by an NOS ceutical compounds (e.g., a pharmaceutical compound inhibitor and a nitric oxide scavenger, i.e., peroxynitrite that is therapeutically active against ALS). Administration and hydroxyl radical scavenger. of a bile composition of the disclosure with another phar- [0019] S-nitrosoglutathione (GSNO) may be a useful 35 maceutical compound may, in some embodiments, (a) NO donor that may slowly and spontaneously release increase the intensity of a response to the pharmaceuti- NO under physiological conditions. GSNO may be a stor- cal compound, (b) increase the efficacy of the pharma- age and/or transport vehicle for NO in the body. A met- ceutical compound, (c) decrease the required dose of abolic enzyme for GSNO may be conserved from bacte- the pharmaceutical compound, and/or (d) decrease the ria to humans. Endogenous GSNO, which may be gen- 40 toxicity of the pharmaceutical compound. Solutions of erated in endothelial cells and astroglial cells during ox- the disclosure may also be administered separately, in idative stress, may be located in the cerebellum in rats. terms of both the route and time of administration. Thus, GSNO may be an endogenous NO reservoir and [0024] A solution of the disclosure may be used, in may play one or more roles in the brain. Interestingly, some embodiments, to treat or ameliorate ALS disease remarkable apoptosis may be observed when cells are 45 and/or advanced ALS disease. For example, a solution treated with GSNO. of the disclosure may include a pharmaceutical com- [0020] Some of the FALS mutant CuZnSOD enzymes pound that decreases motor neuron death such as Pas- may induce a significantly increased peroxidative activity iniazide (Tuberculostatic), Benzthiazide (Diuretic, antihy- in comparison to the wild type protein in vitro. Peroxyni- pertensive), Prednisolone (Glucocorticoid), Menthol 50 trite, a product of superoxide (O2) and nitric oxide (NO), Topical analgesic, (antipuritic), Mebhydrolin Naphthale- reacts with the Cu2+ of mutant SODs, producing nitron- nesulfonate (H1, antihistamine), Trichlormethiazide (Di- ium ions, which lead to nitration of proteins and subse- uretic, antihypertensive), Oxytetracycline (Antibacterial), quent neurotoxicity; motor neurons of ALS patients ex- Arcaine sulphate (NOS inhibitor, NMDA inhibitor, anti- hibit increased immunoreactivity for nitrotyrosine. The protozoal), Erythromycin (Antibacterial), Glutathione enhanced peroxidase activity may increase production 55 (Heavy metal poisoning, antioxidant), Trioxsalen (Mela- of hydroxyl radicals, which could damage neurons. nizing agent, antipsoriatic), NylidrinHCL (Peripheral va- [0021] In some embodiments, a bile acid composition sodilator), Desmethyldiazepam (Sedative, minor tranqui- of the disclosure may lack one or more of the disadvan- lizer), Thonzylamine HCL (Antihistamine), Valproate (Na

5 7 EP 2 255 812 A1 8

Anticonvulsant), Aminophenazone(Antipyretic, analge- sodeoxycholic acid (UDCA) may inhibit hepatocyte ap- sic), Sulfamethizole (Antibacterial), Droperidol (Neu- optosis in vivo. Both in hepatocytes and in nonhepatic roleptic), 2-Thiouracil Thyroid (depressant), Kynurenic cells apoptosis may be induced with various factors such acid (Nutrient in vitamin B deficiency diseases), Fusidic as hydrophobic acids, ethanol, transforming growth fac- acid(Antibacterial), Leucovorin Ca (Anti-anemic, anti- 5 tor-α, an agonistic Fas antibody, or okadaic acid. Sur- dote to folic acid antagonists), Sparteine sulfate (Oxyto- prisingly, UDCA may attenuate apoptosis and display cy- cic), Amygdalin (Anti-inflammatory, experimental antin- toprotection by modulating mitochondrial membrane per- eoplastic), Pramoxine HCL (Anesthetic; topical), Furo- turbation, Bax translocation and/or cytochrome c re- semide (Diuretic, antihypertensive), Dinitolmide (Anti- lease. protozoal), Budesonide(Anti-inflammatory), Flopropione 10 [0026] Ursodeoxycholic acid (3K-7K-dihydroxy-5K- (Antispasmodic), Fluorometholone(Glucocorticoid, anti- cholanic acid; UDCA) is a non- toxic hydrophilic bile acid inflammatory), Formylmethionylphenylalanine N- and normally present in human bile, albeit in a low con- (Chemotactic peptide), Thiopental Na (Anesthetic), Lan- centration of only about 3% of total bile acids. UDCA may soprazole (Antiulcerative), Bretylium Tosylate (Inhibitor be used for the treatment of various cholestatic disorders of norepinephrine release), Cefamandole Na (Antimicro- 15 for which it is the only drug approved by the U.S. Food bial), Oxybendazole (Antihelmintic), Cycloleucylglycine and Drug Administration (FDA). (Inhibits narcotic-induced dopamine R sensitivity), Dan- [0027] A major component of bear bile, UDCA may be trolene Na (Skeletal muscle relaxant), Tetroquinone useful as a pharmaceutical agent for the treatment of and (Keratolytic), Piperazine (Antihelmintic), Aesculin (Anti- the protection against many types of liver disease. Its inflammatory), Ethisterone(Progestin), Dimethadione 20 medicinal uses at the present day include the dissolution (Anticonvulsant), Griseofulvin (Antifungal, inhibits mito- of radiolucent gall stones and various cholestatic disor- sis in metaphase, interacts with, polymerized microtu- ders which are primary biliary cirrhosis, primary scleros- bules and associated proteins), Acetaminosalol (Analge- ing cholangitis, intrahepatic cholestasis of pregnancy, sic, antipyretic), Isoguvacine HCL (GABA agonist), Pu- cystic fibrosis- associated liver disease, a number of pedi- trescine DIHCL (Ornithine decarboxylase inhibitor, cell 25 atric liver disorders, and chronic graft-versus-host dis- growth factor), Emetine HCL (Antiamebic, inhibits RNA, ease of the liver. DNA, and protein synthesis), Sulfanilamide (Antibacteri- [0028] Pharmacological action of UDCA may include al), Mimosine (Depilatory agent), Acetylcholine (Cardiac replacement and/or displacement of toxic bile acids depressant, miotic, peripheral vasodilator), Pralidoxime through UDCA in a dose-dependent manner, cytoprotec- Mesylate (Cholinesterase reactivator), LysylTryptopha- 30 tive effects in a dose-dependent manner, stabilization/ nyl-Lysine (acetate Binds to DNA), Hecogenin (Steroid protection of cell membranes in a dose- dependent man- precursor), Prednisolone acetate (Glucocorticoid), Al- ner, antiapoptotic effects in a dose-dependent manner, bendazole (Antihelmintic), Hydrochlorothiazide (Diuret- immunomodulatory effects due to activation of the intra- ic), Demeclocycline HCL (Antibacterial), Nitrofurazone cellular glucocorticoid receptor in a dose-dependent (Topical anti-infective), Dicloxacillin Na (Antibacterial), 35 manner, antiinflammatory effects due to repression of alpha-Tocopherol (Vitamin E deficiency), Tetracycline NF-kB and inhibition of the induction of nitric oxide syn- HCL (Anti-amebic, antibacterial, antirickettsial), Fenofi- thase, stimulation of bile secretion in a dose-dependent brate (Antihyperlipemic), Probenecid (Uricosuric), manner, Stimulation of exocytosis and insertion of canal- Tretinoin (Keratolytic), Acetaminophen (Analgesic, anti- icular membrane transporters in a dose- dependent man- pyretic), Hydrastinine HCL (Cardiotonic, uterine homeo- 40 ner. static), d[-Arg-2]Kyotorphin acetate (Analgesic), NMDA [0029] UDCA is practically insoluble at pH 1 to 8. The NMDA agonist), Cefmetazole Na (Antimicrobial), Riba- solubility of its protonated form is about 0.05mM. The virin (Antiviral), O-Benzyl-L-Serine (Amino acid deriva- solubility of its taurine conjugated metabolite (TUDCA; tive), Picrotoxin (Stimulant, convulsant, GABA R antag- 0.45mM) is about ten times higher than UDCA solubility. onist, ichthyotoxin), Oxethazine (Local anesthetic), Sul- 45 Moreover, TUDCA is the only bile acid (BA) with relatively fathiazole (Antibacterial), Trichlormethine (Antineoplas- low solubility when protonated. Following oral adminis- tic, cytotoxic), Nabumetone (Anti-inflammatory), Chlo- tration, approximately 30 to 60% of UDCA is absorbed ramphenicol (Antibacterial, antirickettsial, inhibits protein along the length of the jejunum and ileum by nonionic synthesis), riluzole, ginseng and its extract, glycyrrhizin passive diffusion and is absorbed in the ileum by active and glycyrrhizic acid, derivatives of carboquinone, coen- 50 transport mechanisms and to a small extent (20% of an zyme Q10, creatine, insulin-like growth factor-1, minoc- ingested dose) in the colon due to the insolubility of crys- ycline, mecamserin, xaliproden, gabapentin, dex- tal UDCA, which causes extremely slow and incomplete tromethorphan, talampanel, IL-1, TR-500, procysteine, dissolution due to the low aqueous solubility of its non- brain derived neurotrophic factor, baclofen, tizanidin, ionized molecules and more lipophilicity than the ionized benzodiazepines, glycopyrrolate, atropine, quinine,55 bile salt species, and can therefore partition into biolog- phenytoin and morphine. ical membranes. [0025] Hydrophobic bile salts fed to rats may induce [0030] Once taken up by hepatocytes, UDCA may be apoptosis in the liver. In addition, coadministration of ur- conjugated to TUDCA and GUDCA, the latter two being

6 9 EP 2 255 812 A1 10 the secreted bile acids in humans and excreted in bile levels is suggested during the first year of riluzole treat- by hepatic first-pass clearance. Consequently, its blood ment. levels are extremely low in the systemic circulation. Bile [0035] Bile acids may act as intracellular signaling acids undergo extensive hepatic recycling, or free UDCA agents, which modulate cellular transport, alter intracel- may also be secreted by hepatocytes in bile, where it 5 lular Ca2+ levels, and activate cell surface receptors. Ur- may be actively and efficiently reabsorbed by cholangi- sodeoxycholic acid (UDCA) is a hydrophilic bile acid with ocytes. UDCA and GUDCA are absorbed by both active proven clinical efficacy in the treatment of hepatobiliary and passive transport mechanisms, while tauro-conju- disorders. UDCA may be rapidly conjugated with glycine gated UDCA (TUDCA) may be transported actively in the or taurine in vivo to produce glycoursodeoxycolic and terminal ileum. 10 tauroursodeoxycholic (TUDCA) acids, respectively. UD- [0031] In some embodiments, a UDCA dose above CA and its derivatives and conjugates may function as 1012 mg/kg per day may not further increase its pro- cytoprotective agents by inhibiting apoptosis. portion in bile since a large quantities of UDCA may be [0036] Since glutamate neurotoxicity may result in cell biotransformed to CDCA through 7-keto-lithocholic acid death through apoptosis, blocking apoptosis may slow by intestinal bacteria. Alternatively, UDCA may be con- 15 acute and chronic neurodegenerative processes. In verted to CDCA by epimerization of theβ -hydroxyl 7 some embodiments of the disclosure, a bile composition group and further to lithocholic acid (LCA). Therefore, blocks a toxic effect mediated by p53. In some embodi- with increasing doses of UDCA the absorption of UDCA ments of the disclosure, a bile composition blocks a toxic decreases. effects mediated by an oxidative process. [0032] In some embodiments, administration of a com- 20 [0037] The present disclosure relates to an aqueous position of the disclosure may achieve adequatesolution comprising (i) one or more soluble bile acids, amounts of UDCA in the liver, in the systemic circulation, aqueous soluble bile acid derivatives, bile acid salts, or and/or in brain to have a therapeutic effect. A solution of bile acid conjugated with an amine, (collectively "bile ac- the disclosure may, in some embodiments, display sig- id"), (ii) water, and (iii) one or more aqueous soluble nificantly increased aqueous solubility of UDCA, in-25 starch conversion products or aqueous soluble non- creased membrane permeability, protection from starch polysaccharides in an amount sufficient to pro- epimerization of UDCA to CDCA. duce a solution which does not form a precipitate at any [0033] Survival time and quality of life for ALS patients pH within a desired pH range. The composition may con- may be improved by respiratory therapy given while tain a bile acid or a bile acid salt which itself has phar- sleeping during early stages of the disease and alterna- 30 maceutical effectiveness. Formulations of the disclosure tive feeding methods that maintain good nutrition as the may act as a carrier, an adjuvant or enhancer for the disease progresses and swallowing becomes more dif- delivery of a pharmaceutical material which remains dis- ficult. To date, only one drug, riluzole, has been approved solved in the composition of the disclosure across the by the U.S. Food and Drug Administration for treatment desired pH range. Alternatively, according to some em- of ALS. However, the lifespan of patients receiving rilu- 35 bodiments of the disclosure, the composition may com- zole is only extended by a few months. Research related prise a non-bile acid pharmaceutical that is incompletely to stem cells and gene therapy are promising frontiers, soluble. but have not yet enhanced the options available to treat- [0038] In some embodiments, it may be an advantage ing physicians. of this disclosure that the bile acid and the carbohydrate [0034] Without being limited to any particular mecha- 40 remain in solution without precipitation at any pH from nism of action, the present disclosure provides clear, sta- acidic to alkaline. These aqueous solution systems of ble solutions of soluble bile acids that may ameliorate a bile acid are substantially free of precipitate or particles. neurodegenerative process. In some embodiments of A further advantage of this disclosure is that the aqueous the disclosure, the compositions comprise riluzole. Rilu- solution systems demonstrate no changes in physical zole, the only drug for treating ALS to yet receive FDA 45 appearance such as changes in clarity, color or odor fol- approval may function by reducing the amount of gluta- lowing the addition of strong acids or alkali even after mate released during signal transduction. Riluzole effi- several months observation under accelerated condi- cacy has been demonstrated primarily in two principal tions of storage at 50°C. controlled clinical trials. The drug’s most frequent ad- [0039] In some embodiments of the disclosure, an verse events were nausea, vomiting, anorexia, diarrhea, 50 aqueous solution system of a bile acid is administered asthenia, somnolence, vertigo, circumoral paresthesia, orally whereupon it reaches the intestine through the gas- abdominal pain and dizziness. Of these, vertigo, di- trointestinal track without precipitation of bile acids by arrhea, nausea, circumoral paresthesia and anorexia ap- exposure to acidic gastric juices and alkaline juices of pear more frequently in patients that received higher dos- the intestine.These dissolved bile acid formulationsdem- es. Increased serum transaminase levels have generally 55 onstrate intact solution systems in the intestine can be been observed within three months of starting riluzole effectively and completely absorbed and, consequently, treatment. however, these levels recede after two to six undergo enterohepatic cycling. According to an embod- months of treatment. Monitoring serum transaminase iment of the disclosure, bile acid solubility (e.g. precipi-

7 11 EP 2 255 812 A1 12 tation and changes in physical appearance) is affected id, hyodeoxycholic acid, deoxycholic acid, 7-oxolitho- by whether a carboxylic acid side chain of certain bile cholic acid, lithocholic acid, iododeoxycholic acid, iochol- acids can be protonated (non- ionized), is ionized, or is a ic acid, tauroursodeoxycholic acid, taurochenodeoxy- simple carboxylic acid. cholic acid, taurodeoxycholic acid, taurolithocholic acid, [0040] The ionization state of a bile acid carboxylic acid 5 glycoursodeoxycholic acid, taurocholic acid, glycocholic side chain may greatly affect the hydrophobicity and the acid, and their derivatives at a hydroxyl or carboxylic acid hydrophillicity of the bile acid in some aqueous solution group on the steroid nucleus. systems. In some embodiments of the disclosure, that [0044] In some embodiments of the disclosure, one ionization state is manipulated by adjusting the pH to advantage may be that delivery of bile acid in solution control the toxicity, absorption, and amphiphilicity of bile 10 achieves higher in vivo levels of bile acids than existing acids. One or more bile acids may be dissolved in these commercial preparations. Therefore, the therapeutic po- aqueous solution systems as a therapeutically active tential of bile acid may be more fully achieved than pre- agent, as an adjuvant of a drug, as a carrier of a drug or vious formulations. The in vivo levels of bile acids attain- as an enhancer of drug solubility. These aqueous solu- able with existing formulations inwhich bile is incomplete- tion systems may be prepared for oral consumption, en- 15 ly solubilized are lower and require administration of larg- emas, mouthwashes, gargles, nasal preparations, otic er amounts of bile acids. Since bile acid is completely preparations, injections, douches, topical skin prepara- dissolved in the inventive formulations, higher in vivo lev- tions, other topical preparations, and cosmetic prepara- els of bile acid may be achieved, even though lower dos- tions which have a desired pH without the disadvantage es are administered. of precipitation or deterioration in physical appearance 20 [0045] In some embodiments of the disclosure, a plu- after long periods of time. rality of bile acids may be used in a single formulation. [0041] Soluble bile acids are any type of aqueous sol- Mixtures of two or more bile salts of differing hydrophobic uble bile acids. A bile acid salt is any aqueous soluble activity may behave asa single bilesalt of an intermediate salt of a bile acid. Bile salts exhibit greater solubilizing hydrophobic activity. As a result, detergent properties capacity for phospholipid and cholesterol and are con- 25 and the toxicity of mixtures of two bile acids of differing sequently better detergents. More hydrophobic bile salts hydrophobic activity often are intermediate between the may be more injurious to various membranes, both in individual components. vivo and in vitro. Aqueous dissolved salts of bile acids [0046] Mixtures of two or more bile salts of differing may be formed by the reaction of bile acids described hydrophobic activity may behave as a single bile salt of above and an amine including but not limited to aliphatic 30 an intermediate hydrophobic activity. As a result, deter- free amines such as trientine, diethylene triamine, tetra- gent properties and the toxicity of mixtures of two bile ethylene pentamine, and basic amino acids such as ar- acids of differing hydrophobic activity often are interme- ginine, lysine, ornithine, and ammonia, and amino sugars diate between the individual components. such as D-glucamine, N-alkylglucamines, and quater- [0047] Carbohydrates suitable for use in the disclosure nary ammonium derivatives such as choline, heterocyclic 35 include aqueous soluble starch conversion products and amines such as piperazine, N-alkylpiperazine, piperid- aqueous soluble non-starch polysaccharides. According ine, N-alkylpiperidine, morpholine, N-alkylmorphline, tosome embodiments of the present disclosure, aqueous pyrrolidine, triethanolamine, and trimethanolamine. Ac- soluble starch conversion products include carbohy- cording to the disclosure, aqueous soluble metal salts of drates obtained directly from the partial or incomplete bile acids, inclusion compound between the bile acid and 40 hydrolysis of starch under various pH conditions. Non- cyclodextrin and its derivatives, and aqueous soluble O- limiting examples include maltodextrin, dextrin, liquid glu- sulfonated bile acids are also included as soluble bile cose, corn syrup solid (dried powder of liquid glucose), acid salts. and soluble starch, (e.g., maltodextrin or corn syrup sol- [0042] Soluble bile acid derivatives according to some id). In some embodiments, MALTRIN ® M200, a corn syr- embodiments of this disclosure, may be those derivatives 45 up solid, and MALTRIN® M700, a maltodextrin, may be which are as soluble in aqueous solution as or more sol- used and both of which are manufactured by GPC®, uble in aqueous solution than is the corresponding un- Grain Processing Corporation of Muscatine, Iowa may derivatized bile acid. Bile acid derivatives include, but are be used. For the purpose of this embodiment, the term not limited to derivatives formed at the hydroxyl and car- "corn syrup" includes both corn syrup and liquid glucose. boxylic acid groups of the bile acid with other functional 50 If a starch conversion product is polymeric, the polymer groups including but not limited to halogens and amino has at least one reducing end and at least one non-re- groups. Soluble bile acid may include an aqueous prep- ducing end and may be linear or branched. The molecular aration of a free acid form of bile acid combined with one weight may be from about 100 mass units to over 106 of HCl, phosphoric acid, citric acid, acetic acid, ammonia, mass units. High molecular weight aqueous soluble or arginine. 55 starch conversion products are those having a molecular [0043] Bile acids that may be used in accordance with weight over 105. the teachings of this disclosure include, without limitation, [0048] According to some embodiments of the present ursodeoxycholic acid, chenodeoxycholic acid, cholic ac- disclosure, aqueous soluble non- starch polysaccharides

8 13 EP 2 255 812 A1 14 may be under various pH conditions by various hydrolytic and wheat bran. or synthetic mechanisms. Non- limiting examples include [0052] In some embodiments of the disclosure, the for- to dextran, guar gum, pectin, indigestible soluble fiber. If mulation further comprises emulsifying agents. For the polymeric, the polymer has at least one reducing end and purpose of the disclosure, the term "emulsifying agent" at least one non-reducing end. The polymer may be linear 5 includes emulsifying agents and suspending agents. or branched. The molecular weight is from about 100 Non-limiting examples of emulsifying agents include guar mass units to over 106 mass units. Preferably the mo- gum, pectin, acacia, carrageenan, carboxymethyl cellu- lecular weight is over 105 mass units. lose sodium, hydroxymethyl cellulose, hydroxypropyl [0049] The amount of high molecular weight aqueous cellulose, methyl cellulose, polyvinyl alcohol, povidone, soluble starch conversion product and/or soluble non- 10 tragacanth gum, xanthan gum, and sorbian ester. starch polysaccharide used in embodiments of the dis- [0053] The selected pH range for which the formulation closure is at least the amount needed to render the cho- will not precipitate its bile acid, starch conversion product, sen bile acid(s) in the preparation soluble in the concen- soluble non-starch polysaccharide or its pharmaceutical tration desired and in the pH range desired. In some em- compound may be any range of pH levels obtainable with bodiments of the disclosure, the approximate minimal 15 an aqueous system. Preferably this range is between weight ratio of maltodextrin to UDCA required to prevent about pH 1 and about pH 14 and more preferably be- UDCA precipitation is 6:1 (i.e. 1.2 g for every 0.2 g of tween about pH 1 and about pH 10. Still more preferably UDCA, 6 g for every 1g of UDCA, and 12 g for every 2 g the range is any subset of the range of pH levels obtain- of UDCA in 100 mL of water). In some embodiments of able in an aqueous system sufficient for a pharmaceutical the disclosure, the approximate minimal quantity of mal- 20 formulation to remain in solution from preparation, to ad- todextrin is 30 g for every 200 mg of chenodeoxycholic ministration, to absorption in the body, according to the acid, 12 g for every 200 mg of 7-ketolithocholic acid, 10 method of administration. Thus, the composition may be g for every 200 mg of cholic acid and 50 g for every 200 used as a pharmaceutical formulation wherein the phar- mg of deoxycholic acid. In some embodiments of the dis- maceutical compound remains in solution without pre- closure, the approximate minimal weight ratio of liquid 25 cipitation at prevailing pH levels in the mouth, stomach glucose (commercial light corn syrup) to UDCA required and intestines. In some embodiments of the disclosure, to prevent the precipitation of bile acids from the aqueous a bile acid remains dissolved under acidic conditions as solution dosage forms of the disclosure is about 25: 1 (i.e. a free bile acid in spite of the general insolubility of bile 12.5 g for every 500 mg UDCA in 100 mL water and 25 acids under acidic conditions. g for every 1 g ursodeoxycholic acid in 200 mL water). 30 [0054] In some embodiments of the disclosure, the In some embodiments of the disclosure, the approximate pharmaceutical is riluzole. Non-limiting examples of oth- minimal quantity of dried powder of liquid glucose (corn er pharmaceutical compounds include hormones, hor- syrup solid, e.g. MALTRIN® M200) required to prevent mone antagonists, analgesic, antipyretics, anti-inflam- the precipitation of bile acids from the aqueous solution matory drugs, immunoactive drugs, antineoplastic drugs, dosage forms of the disclosure is 30 g for every 1 g ur- 35 antibiotics, anti-inflammatory agents, sympathomimetic sodeoxycholic acid in 100 mL water, and approximately drugs, anti-infective drugs, anti-tumor agents, and anes- 60 g for every 2 g of ursodeoxycholic acid in 200 mL thetics. Further non-limiting examples include drugs that water. In some embodiments of the disclosure, the ap- target or affect the gastrointestinal tract, liver, cardiovas- proximate minimal quantity of soluble starch non- cular system, and respiratory system. Further non-limit- polysaccharide required to prevent the precipitation of 40 ing examples of pharmaceutical compounds include in- bile acids from the aqueous solution dosage forms of the sulin, heparin, calcitonin, ampicillin, octreotide, sildenafil disclosure is 50 g guar gum for every 500 mg ursodeox- citrate, calcitriol, dihydrotachysterol, ampomorphine, yo- ycholic acid in 100 mL water and 80g of pectin for every himbin, trazodone, acyclovir, amantadine •HCl, rimanta- 500 mg of ursodeoxycholic acid in 100 mL water. The dine•HCl, cidofovir, delavirdine•mesylate, didanosine, minimal required quantity of high molecular weight aque- 45 famciclovir, forscarnet sodium, fluorouracil, ganciclovir ous soluble starch conversion products or soluble non- sodium, idoxuridine, interferon-α, lamivudine, nevirap- starch polysaccharide is primarily determined by the ab- ine, penciclovir, ribavirin, stavudine, trifluridine, valacy- solute quantity of bile acids in the solution formulation clovir•HCl, zalcitabine, zidovudine, indinavir•H2SO4, rather than the concentration. ritonavir, nelfinavir•CH3SO3H, saquinavir•CH3SO3H, d- [0050] In some embodiments of the disclosure, a for- 50 penicillamine, chloroquine, hydroxychloroquine, aurothi- mulation may comprise cyclodextrin in addition to a oglucose, gold sodium thiomalate, auranofin levamisole, starch conversion product and/or a non-starch polysac- DTC, isoprinosine, methyl inosine monophosphate, mu- charide. Alternatively, in some embodiments, a compo- ramyl dipeptide, diazoxide, hydralazine•HCl, minoxidil, sition of the disclosure may lack cyclodextrin. dipyridamole, isoxsuprine•HCl, niacin, nylidrin•HCl, 55 [0051] In some embodiments of the disclosure, the for- phentolamine, doxazosin•CH3SO3H, prazosin•HCl, ter- mulation further comprises dietary fiber. Non- limiting ex- azocin•HCl, clonidine•HCl, nifedipine, molsidomine, ami- amples of dietary fiber include guar gum, pectin, psyllium, odarone, acetylsalicylic acid, verapamil, diltiazem, nisol- oat gum, soybean fiber, oat bran, corn bran, cellulose dipine, isradipine, bepridil, isosorbide•dinitrate, pentaer-

9 15 EP 2 255 812 A1 16 ythrytol•tetranitrate, nitroglycerin, cimetidine, famotidine, catechin and its derivatives, glycyrrhizal extract and its nizatidine, ranitidine, lansoprazole, omeprazole, misopr- main constituents such as glycyrrhizin and glycyrrhizic ostol, sucralfate, metoclopramide•HCl, erythromycin, acid, water soluble bismuth compounds e.g( ., bismuth bismuth compound, alprostadil, albuterol, pirbuterol, sodium tartrate), and which are soluble and stable in acid 5 terbutaline•H2SO4, salmetrol, aminophylline, dyphylline, and/or alkali can be added as needed into these aqueous ephedrine, ethylnorepinephrine, isoetharine, isoprotere- solution dosage formulations containing ursodeoxychol- nol, metaproterenol, n- docromil, oxytriphylline, theophyl- ic acid in this disclosure. line, bitolterol, fenoterol, budesonide, flunisolide, be- [0056] Some embodiments of the disclosure may be clomethasone•dipropionate, fluticasone•propionate, co- practiced with pH adjustable agents. Non- limiting exam- 10 deine, codeine sulfate, codeine phosphate, dextrometh- ples include HCl, H 3PO4, H2SO4, HNO3, CH3COOH, cit- orphan•HBr, triamcinolone•acetonide, montelukast sodi- ric acid, malic acid, tartaric acid, lactic acid, phosphate, um, zafirlukast, zileuton, cromolyn sodium, ipratropium eidetic acid and alkalies. bromide, nedocromil sodium benzonate, diphenhy- [0057] In some embodiments of the disclosure, the for- dramine•HCl, hydrocodone•bitartarate, methadone•HCl, mulations may be used to treat human and mammalian morphine sulfate, acetylcysteine, guaifenesin, ammoni- 15 diseases. The disclosure contemplates treating ALS, um carbonate, ammonium chloride, antimony potassium ALS-related disorders, and other neurodegenerative dis- tartarate, glycerin, terpin•hydrate, colfosceril palmitate, orders. Solutions of the disclosure may also be used to atorvastatin•calcium, cervastatin•sodium, fluvastatin•so- treat gastrointestinal disorders, liver diseases, gall dium, lovastatin, pravastatin•sodium, simvastatin, picror- stones, and hyperlipidemia. Non- limiting examples of liv- rhazia kurrva, andrographis paniculata, moringa oleifera, 20 er diseases include alcohol-induced liver diseases and albizzia lebeck, adhata vasica, curcuma longa, momor- non-alcohol-induced liver diseases. Non-limiting exam- dica charantia, gymnema sylvestre, terminalia arjuna, ples of gastrointestinal disorders include chronic gastri- azadirachta indica, tinosporia cordifolia, metronidazole, tis, reflux gastritis, and peptic ulcer disease. Non- limiting amphotericin B, clotrimazole, fluconazole, haloprogin, examples of non-alcohol-induced liver diseases include ketoconazole, griseofulvin, itraconazole, terbinafin•HCl, 25 primary biliary cirrhosis, acute and chronic hepatitis, pri- econazole•HNO3, miconazole, nystatin, oxicona- mary sclerosing cholangitis, chronic active hepatitis, and zole•HNO3, sulconazole•HNO3, cetirizine•2HCl, dexam- excess accumulation of fat in the liver. The disclosure ethasone, hydrocortisone, prednisolone, cortisone, cat- further contemplates treating viral, bacterial and fungal echin and its derivatives, glycyrrhizin, glycyrrhizic acid, diseases. In some embodiments of the disclosure, a for- betamethasone, ludrocortisone•acetate, flunisolide, flu- 30 mulation is administered to treat and/or eradicate Heli- ticasone•propionate, methyl prednisolone, somatostatin, cobacter pylori infection. In some embodiments of the lispro, glucagon, proinsulin, insoluble insulins, acarbose, disclosure, a formulation is administered to treat and/or chlorpropamide, glipizide, glyburide, metformin•HCl, eradicate hepatitis C virus infection, influenza A, Influen- repaglinide, tolbutamide, amino acid, colchicine, sulfin- za C, parainfluenza 1, sendai, rubella, and pseudorabies pyrazone, allopurinol, piroxicam, tolmetin sodium, in-35 virus. In some embodiments of the disclosure, a formu- domethacin, ibuprofen, diflunisal, mefenamic acid, lation is administered to treat acute or chronic inflamma- naproxen, and trientine. tory diseases. Non-limiting examples of inflammatory [0055] Additional examples of pharmaceutical com- diseases include bronchitis, chronic pharyngitis, and pounds that may be included in the formulation are any chronic tonsillitis. In some embodiments of the disclo- compounds which remain soluble when added to the for- 40 sure, a formulation is administered to treat hypercholer- mulation. With an additional pharmaceutical compound sterolemia. in the formulation, a bile acid in solution may act as an [0058] In some embodiments of the disclosure, the for- adjuvant, carrier, or enhancer for the solubility of certain mulation is modified such that it may be administered as therapeutically active agents, including, but not limited a liquid, solid, powder or tablet. In some embodiments to, insulin (pH 7.4-7.8), heparin (pH 5-7.5), calcitonin, 45 of the disclosure, the formulation is comprised in a ampicillin, amantadine, rimantadine, sildenafil, neomycin parenteral solution (e.g., an injectable solution, a solu- sulfate (pH 5-7.5), apomorphine, yohimbin, trazodone, tion, a syrup, a thick syrup or a paste. A non- limiting ex- ribavirin, paclitaxel and its derivatives, retinol, and tretin- ample of a syrup is a solution of maltodextrin wherein the oin, which are soluble and stable in acid and/or alkali and concentration of maltodextrin is less than 500 g/L. A non- can be added as needed into these aqueous solution 50 limiting example of a syrup is a solution of maltodextrin dosage forms of certain concentrations of bile acids in wherein the concentration of maltodextrin is between 500 this disclosure. Certain therapeutically active agents, in- g/L and 1.0 kg/L inclusive. A non-limiting example of a cluding, but not limited to, metformin HCl (pH 5-7), rani- thick syrup is a solution of maltodextrin wherein the con- tidine HCl, cimetidine, lamivudine, cetrizine 2HCl (pH centration of maltodextrin is between 1.0 kg/L and 1.2 4-5), amantadine, rimantadine, sildenafil, apomorphine, 55 kg/L inclusive. A non-limiting example of a paste is a so- yohimbine, trazodone, ribavirin and dexamethasone, hy- lution of maltodextrin wherein the concentration of mal- drocortisone, prednisolone, triamcinolone, cortisone, ni- todextrin is greater than 1.2 kg/L. acin, taurine, vitamins, naturally occurring amino acids, [0059] The stability of dosage formulations of the dis-

10 17 EP 2 255 812 A1 18 closure may be evaluated by measuring the concentra- evaluated in like manner. The highest cell viability (87% tion of the relevant bile acid over time in preparations compared to control, 100%) was observed in cells treated comprising soluble bile acid, a high molecular weight withboth cisplatin (20 PM)and a solution of the disclosure aqueous soluble starch conversion product, and water (1mg/ml soluble UDCA), whereas the lowest cell viability at various pH and temperature levels. The retention time 5 (35% compared to control, 100%) was observed in cells (high performance liquid chromatography) of each bile treated with cisplatin (20 PM) alone. These effects were acid may be adjusted as needed to permit individual anal- also found in a dosage- dependent fashion. According to ysis each bile acid present in complex samples, i.e. a the MTT assays, a solution of the disclosure may block sample having a plurality of bile acids. Stability tests may almost completely hydrogen peroxide- induced oxidative also be performed by assessing the light-scattering prop- 10 cytotoxicity and may completely block cisplatin-induced erties of a test solution. In addition, established acceler- oxidative cytotoxicity. In conclusion, solution of the dis- ated testing conditions may be used. closure possess a strong antioxidative properties and [0060] All stability tests performed on solutions of the non-cytotoxicity. disclosure were satisfactory in that the concentration of bile acid as measured by HPLC did not change appreci- 15 EXAMPLES ably over time at various pH levels. Particularly, all bile acid solution formulations tested showed excellent re- [0064] Some embodiments of the present disclosure sults in the stability tests with no precipitation and no may be understood in connection with the following ex- physical appearance changes over the test period. Some amples. However, one skilled in the art will readily ap- formulations remain stable for over 2 years. The aqueous 20 preciate the specific materials, compositions, and results solution dosage forms according to this disclosure that described are merely illustrative of the disclosure, and weretested did not change either physically or chemically are not intended to, nor should be construed to, limit the atvarious pH conditions under the accelerated conditions scope disclosure and its various embodiments. despite the addition of therapeutically and chemically active agents that are stable and soluble in hydrochloric 25 Example 1: Preparation of Bile Acid Solutions acid solution. Therefore, these aqueoussolution systems may be extremely valuable pharmaceutical dosage [0065] A stock solution of bile acid was prepared by forms for the therapeutically active bile acids prepara- first dissolving UDCA (60 g) in 500 mL NaOH (6.7 g) tions, and/or the drug (pharmaceutical compound) deliv- solution. Next, to the resulting clear solution, 375 g of ery preparations in which bile acids play roles as the ad- 30 maltodextrin was added, portion by portion with vigorous juvant of drug, the carrier of drug, or the enhancer of agitation. The pH was then adjusted to between 7.0 and solubility of a drug by micelle formation at various pH 7.2 by the dropwise addition of HCl with high throughput conditions without the stability problems, including pre- sonication (750W, 20kHz). The volume was then adjust- cipitation in acidic conditions. ed to 1.0 L with injectable distilled water. Lastly, the re- [0061] Human neuronal cells were treated with a so- 35 sulting clear solution was filtered with sterilized using a lution of the disclosure and 50 PM of hydrogen peroxide 0.22 P GP express plus membrane under aseptic con- and/or cisplatin. Hydrogen peroxide is strong oxidant. ditions. (This filtration is important for sterilization, but not Cisplatin stimulates production of reactive oxygen spe- for removing particulate matter since the solution is al- cies (ROS), which interfere with the antioxidant defense ready clear.) Dilutions of this solution to the desired UD- system. Cell viability, cell proliferation, and apoptosis 40 CA concentration were prepared according to standard were then analyzed by measurement of MTT reduction. pharmacy practice. Several studies, using exogenous ROS, and 2HO2, in particular, demonstrate that exposure of human and rat Example 2: Preparation of Bile Acid Solutions peripheral vascular smooth muscle cells (VSMCs) to relatively low levels of oxidant stress, for short periods pro- 45 [0066] A stock solution of bile acid was prepared by motes cell growth, whereas prolongedexposure to higher first dissolving UDCA (25 g) in 400 mL NaOH (2.7 g) concentrations leads to cell death, either by apoptosis or solution. Next, to the resulting clear solution, 745 g of necrosis. maltodextrin was added, portion by portion with vigorous [0062] Cell viability for hydrogen peroxide with and agitation. To this resulting solution 100 mL of a preserve without solution of the disclosure was evaluated by using 50 solution which contains 0.95g of methyl p-hydroxyben- the MTT assay. Cells treated with a solution of the dis- zoate and 0.3g of sodium hydrogensulfite was added and closure (0.2 mg/ml solubilized UDCA) and hydrogen per- then stirred. The volume was then adjusted to 1.0 L with oxide (50 PM) displayed the highest cell viability (75% pharmaceutical grade water. Lastly, the resulting clear compared to control, 100%). The lowest cell viability solution was filtered with proper filtering apparatus. (This (26% compared to control, 100%) was observed in cells 55 filtration is not performed to remove particulate matter treated with hydrogen peroxide (50P M) alone. These since the solution is already clear.) Dilutions of this so- effects were found in a dosage-dependent fashion. lution to the desired UDCA concentration were prepared [0063] Cell viability in the presence of cisplatin was according to standard pharmacy practice.

11 19 EP 2 255 812 A1 20

Example 3: Animal Test recovering motor performance.

[0067] Transgenic Rat; The transgenic animals used Example 4: Cell survival experiments in this example were heterozygotic hSOD1 carriers with a glycine93-alanine mutation (G93A). The strain is reg- 5 [0073] To evaluate the protective effect of solubilized istered as B6SJL-TgN(SOD1-G93A)1Gur (The Jackson UDCA solution on wild type cells, G93A cells, and A4V Lab., Bar Harbor, ME, USA) containing a reduced copy cells, these cells were treated with solutions containing number of hSOD1.were purchased from the Jackson 200nM or 20 PM of solubilized UDCA after neuronal dif- Laboratories. Transgenic mice, over-expressing the hu- ferentiation and then, were incubated for 24 h. The pro- man SOD1 gene with a mutation identified in ALS pa- 10 tective effect of solubilized UDCA solution on wild type tients, develop an adult-onset, progressive motor dete- cells, G93A cells, and A4V cells was determined by using rioration. These transgenic mice are considered to be a both the MTT assay and Trypan blue staining. Cell via- model for the disease and have been used to test a bility may be expressed as a percentage of cell survival number of strategies to delay disease progression and as shown in Figure 2. mortality. 15 [0074] GSNO has been known to be associated with [0068] In order to evaluate the potential benefit of cell apoptosis. To evaluate the anti-apoptotic effect of present invention in ALS, 240mg/kg solubilized UDCA in solubilized UDCA against GSNO on wild type cells and the present solution were given by the oral administration cells containing G93A and A4V mutations in hSOD1, twice per week, beginning when G93A transgenic mice wildtype cells, G93A cells, and A4V cells were incubated were 70 days old and continuing until death. 20 with 500 PM GSNO for 24h in order to induce apoptosis [0069] Life expectancy and its results; The mean and then cell viability of was determined. These apopto- life expectancy of the control mice (n=8) and the treated sis-induced wildtype cells, G93A cells, and A4V cells mice (n=8) is shown in Fig. 1. The clear solution (25 were next incubated with 20 PM of solubilized UDCA for mg/kg) increased the mean survival time from 134.8 days 24h and cell viability was determined again using both in the control mice (n=8) to 146.6days in the treated mice 25 the MTT assay and Trypan blue staining. Cell viability (n=8), a delay of mortality with 13 days. This corresponds may be expressed as a percentage of cell survival as to an 8.81% increase in life span of the G93A mice. shown in Figure 3. [0070] Rotarod Test and its results; A rotarod was [0075] Cell culture: VSC 4.1. (ventral spinal cord 4.1; used to evaluate motor performance. Mice were placed motoneuron-neuroblastoma hybrid cells) were main- on the rod against the direction of rotation, forcing them 30 tained in Dulbecco’s modified Eagles’ medium/F-12 to keep moving forwards to avoid slipping off the rod growth medium (Gibco, Grand Island, NY) with Sato’s backwards. After a learning period of several days, mice components (Sigma, St. Louis, MD) and 2% heat-inacti- were able to stay on the rotarod rotating at 15 r.p.m. Mice vated newborn calf serum (HyClone, Logan, UT). They were tested once a week. Each mouse was given three were grown in log-phase growth on poly-(L-orni- trials on each rotarod test and the time they remained on 35 thine)-precoated culture dishes (Falcon, Franklin lakes, the rod before sliding off was recorded (latency). The NJ). After confluency, the cells were plated in 96-well highest staying time on the rotarod was chosen from plates (NUNC, Denmark) at a density of 1 X 10n (n=4) among the three trials as a measure for motor perform- cells/well. For immunoblot, 1 X 10 n (n=5) cells were seed- ance. ed in 100 mm dishes (Falcon, Franklin Lakes, NJ). [0071] The motor performance of treated mice did im- 40 [0076] Constructs and establishing stable cell prove greatly as the disease progressed, while the motor line: Wild type (control) or mutant (G93A, A4V) human performance of untreated mice declined more and more Cu/Zn SOD-1 cDNA was cloned into the BamHI and rapidly as the disease progressed. Differences in motor EcoRII sites of pcDNA 3.0. An empty vector that did not performance between untreated and treated mice was contain the insert was used for the control. Cells were highly significant on day 112 (p=0.01), on day 119(0.001), 45 tranfected (Superfect, Qiagen, Valencia, CA) and main- on day 126(p=0.01), on day 133(p=0.001) and on day tained in a medium that contained G418 at a concentra- 140(p=0.043). tion of 400 mg/ml (Gibco, Grand Island, NY). Single or [0072] Conclusion; 70 days old, an age at which pro- pooled colonies were used for the experiment after clar- gressive cytoskeletal alterations extensively began to ifying the expression of human SOD1 (WT, Mutant) by demonstrate in SOD1- G93A rat, has been known as clin- 50 Western blot analysis using an anti-human SOD1 poly- ical onset of ALS. These data demonstrate that solubi- clonal antibody (Calbiochem, La Jolla, CA). These cell lized UDCA solution significantly prolongs life span, re- lines were grown under the same conditions as the VSC tards the onset of paralysis and slows the evolution of 4.1 cells. functional parameters connected with muscle strength in [0077] MTT assay and trypan blue staining to eval- the ALS-mouse model. Moreover, the improved motor 55 uate the protective effects and its results; 3-(4,5- performance on older models than 70 days old with ad- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide ministration of solubilized UDCA has been provided that (MTT) is absorbed into cells and converted to formazan this invent solution can treat advanced ALS disease with by the action of mitochondrial succinate dehydrogenase.

12 21 EP 2 255 812 A1 22

Therefore, accumulation of formazan reflects the activity undergoing apoptosis in G93A and A4V cells provided of mitochondria directly and the cell viability indirectly. evidence that apoptotic effect of GSNO was more signif- Thus, neuronally differentiated cells (wildtype cells(con- icant on G93A cells and A4V cells than the wild type cells. trol), G93A cells, A4V cells) were plated at a density of [0081] MTT assay and trypan blue staining to eval- 1 X 10n (n=4) cells/well in a 96 well plate. To evaluate 5 uate the anti-apoptotic effects of solubilized UDCA the effect of G93A or A4V mutations on viability, the plat- and its results; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphe- ed cells were incubated with the culture media containing nyltetrazolium bromide (MTT) is absorbed into cells and 1 mM dibutyryl cAMP and 0.025 ug/mL aphidicolin, and converted to formazan by the action of mitochondrial suc- the viability of wildtype cells, G93A cells, and A4V cells cinate dehydrogenase. Therefore, accumulation of for- was evaluated as a function of concentration. To evaluate 10 mazan reflects the activity of mitochondria directly and the protective effect of solubilized UDCA on viability, the cell viability indirectly. Thus, neuronally differentiated 200nM and 20 PM of solubilized UDCA were added into cells (wildtype cells (control), G93A cells, A4V cells) were the media. At 24 h of incubation, 50 PL of 2 mg/mL MTT plated at a density of 1 X 10 n (n=4) cells/well in a 96 well (Sigma, Saint Louis, MO, USA) were added after media plate. The plated cells were incubated with the culture (200 PL) were added into each well. Subsequently, 220 15 media containing 1 mM dibutyryl cAMP and 0.025 ug/mL PL of the solution were removed from each well, and 150 aphidicolin, and then, were further incubated for 24h with/ PL of dimethyl sulfoxide were then added back to each without addition of 500 PM GSNO. To evaluate the anti- well. Finally, optical density (OD) was read at 540 nm on apoptotic effect of solubilized UDCA on cells, 200nM and the ELISA plate reader after particles in the well were 20 PM of solubilized UDCA were added into the media. dissolved by a microplate mixer for 10 min. Results were 20 At 24 h of incubation, 50 PL of 2 mg/mL MTT (Sigma, calibrated using OD measured without cell culture. For Saint Louis, MO, USA) were added after media (200 PL) trypan blue staining, 10 PL of trypan blue solution were were added into each well. Subsequently, 220 PL of the added to 10 PL of cells collected from each plate, and solution were removed from each well, and 150P L of the cells were incubated for 2 min. Unstained live cells dimethyl sulfoxide were then added back to each well. were counted on a haemocytometer. Cell viability was 25 Finally, optical density (OD) was read at 540 nm on the expressed as percentages of the cell survival. ELISA plate reader after particles in the well were dis- [0078] The viability of the wild type cells, G93A cells, solved by a microplate mixer for 10 min. Results were and A4V cells was 100%, 130% and 115%, respectively, calibrated using OD measured without cell culture. For compared to untreated wild type cells (control; 100%), trypan blue staining, 10 PL of trypan blue solution were when treated with 200 nM of solubilized UDCA. At 100 30 added to 10 PL of cells collected from each plate, and times higher concentration of solubilized UDCA (20 PM) the cells were incubated for 2 min. Unstained live cells than 200 nM, the viability of wild type cells, G93A cells, were counted on a haemocytometer. Cell viability was and A4V cells were 89%, 133% and 101%, respectively, expressed as percentages of the cell survival. compared to untreated wild type cells (control; 100%). [0082] The decrement of viability by S-nitrosoglutath- This experimental data showed that solubilized UDCA 35 ione-induced apoptosis was more prominent in G93A has a protective effect on G93A cells and A4V cells, and cells and A4V cells than in wild type cells. The increment is non-toxic to wild type cells, G93A cells and A4V cells. of viability by solubilized UDCA in GSNO treated (apop- [0079] DAPI staining to evaluate apoptosis by totic) cells was significant in G93A cells and A4V cells GSNO and its results; 4’, 6-Diamidino-2-phenylindole compared to the wild type cells. More specifically, com- dihydrochloride (DAPI) staining was performed to eval- 40 pared to the wild type cells (92%), the viabilities of G93A uate apoptosis as follows. After differentiation, the and A4V mutations were 81% and 75%, respectively, at wildtype (control), G93A, and A4V cells were incubated 24 h after the incubation with 500 PM GSNO. After adding with/without 500 PM GSNO for 24 h; the cells were then solubilized UDCA and incubating further, the viability of centrifuged at 265 g for 2 min, and 4% neutral buffered the wild type cells, G93A and A4V cells increased to 98% formalin (100 PL) was added to the cell pellet. A 50 PL 45 from to 92% in the wild type cells, increased to 100% aliquot of the cell suspension was applied to a glass slide from 81% in G93A cells, and increased to 115% from to and dried at room temperature. The fixed cells were 75% in A4V cells. washed in PBS, air dried, and stained for 20 min with the [0083] This experiment demonstrates that soluble UD- DNA-specific fluorochrome, DAPI (Sigma, Saint Louis, CA solution may protect G93A and A4V cell from NO- MO, USA). The adhered cells were rinsed with PBS, air 50 mediated apoptosis and may revive apoptosis-mediated dried, and mounted with 90% glycerol. The slides were damaged cells of ALS. observed under Olympus fluorescence microscopy(pic- [0084] As will be understood by those of ordinary skill tures). Cell viability was expressed as percentages of the in the art, other equivalent or alternative methods, and/or cell survival. compositions for ameliorating and/or treating a symptom [0080] DAPI staining showed that the percent of ap- 55 of a neurodegenerative disease and/or a motor neuron optotic cells among the G93A and A4V cells (19% versus disease. For example, methods and dosages may be 25%, respectively) significantly increased compared with scaled to diagnose and/or treat subjects of different sizes the wild cells (8%). The increased percentage of cells (e.g., children and adults), subjects with additional aller-

13 23 EP 2 255 812 A1 24 gies or conditions, and/or subjects having varying sever- pH values of the solution within a selected ity of allergy and/or symptoms. In addition, methods and range of pH values. dosages may be adapted to fluctuations over time ( e.g., monthly or seasonal). These equivalents and alterna- 2. A method according to claim 1, wherein the neu- tives along with obvious changes and modifications are 5 rodegenerative disease is selected from the group intended to be included within the scope of the present consisting of Parkinson’s disease, Huntington’s dis- disclosure. For example, values and/or range endpoints ease, Alzheimer’s disease, amyotrophic lateral scle- provided are not intended to be rigid limits for all embod- rosis, stroke, and spinal cord injury. iments. Moreover, one of ordinary skill in the art will ap- preciate that no single embodiment, use, and/or advan- 10 3. A method according to claim 2, wherein the neu- tage is intended to universally control or exclude other rodegenerative disease is amyotrophic lateral scle- embodiments, uses, and/or advantages. For example, a rosis. medical practitioner may deem circumstances to warrant giving preference to one over another. While the present 4. A method according to claim 3, wherein the amyo- disclosure includes extensive information about current 15 trophic lateral sclerosis is advanced amyotrophic lat- perceptions of the genetics, biochemistry, and cell biol- eral sclerosis. ogy of ALS and bile acid metabolism, future work may reveal that aspects of these perceptions are inaccurate 5. A method according to claim 1, wherein the symp- or incomplete. Accordingly, as will be understood by tom is selected from the group consisting of short- those skilled in the art, the present disclosure, whether 20 ened lifespan and paralysis. taken in whole or in part, is not limited to a particular model or mechanism of action. In addition, one of ordi- 6. A method according to claim 1, wherein subject nary skill in the art will recognize that other equivalent or is a mammal. alternative compositions and methods may be used. For example, while a number of compounds have been dis- 25 7. A method according to claim 1, wherein subject closed as being capable of administration with a bile acid, is a human. other compounds may be included as well. Also, administration of a pharmaceutical may be performed at the 8. A method according to claim 1, wherein the first same time as administration of a bile acid composition material is present in therapeutically active amount. or the two may simply be administered during the same 30 or overlapping time periods (e.g., during the same hour, 9. A method according to claim 1, wherein the first the same day, or the same week). Accordingly, the fore- material is selected from the group consisting of going disclosure is intended to be illustrative, but not lim- chenodeoxycholic acid, cholic acid, hyodeoxycholic iting, of the scope of the disclosure as illustrated by the acid, deoxycholic acid, 7-oxolithocholic acid, litho- following claims. 35 cholic acid, iododeoxycholic acid, iocholic acid, tauroursodeoxycholic acid, taurochenodeoxycholic ac- CLAUSES id, taurodeoxycholic acid, glycoursodeoxycholic acid, taurocholic acid, glycocholic acid, their deriva- [0085] tives at a hydroxyl or carboxylic acid group on the 40 steroid nucleus, their salts, or their conjugates with 1. A method of ameliorating or treating at least one amines. symptom of a neurodegenerative diseases in a subject comprising: 10. A method according to claim 1, wherein the aqueous soluble starch conversion product is selected administering to a subject a clear aqueous so- 45 from the group consisting of maltodextrin, dextrin, lution comprising: liquid glucose, corn syrup solid, and soluble starch.

(a) a first material selected from the group 11. A method according to claim 1, wherein the se- consisting of a bile acid, an aqueous soluble lected pH range is between approximately 1 and ap- derivative of a bile acid, a bile acid salt, and 50 proximately 10 inclusive. a bile acid conjugated with an amine by an amide linkage; 12. A method according to claim 1, wherein the first (b) a carbohydrate selected from the group material is ursodeoxycholic acid or sodium salt of consisting of an aqueous soluble starch ursodeoxycholic acid. conversion product or an aqueous soluble 55 non-starch polysaccharide; 13. A method according to claim 1, wherein the aque- (c) water, wherein the first material and the ous soluble starch conversion product is maltodex- carbohydrate both remain in solution for all trin.

14 25 EP 2 255 812 A1 26

14. A method according to claim 1, wherein the aque- roursodeoxycholic acid, taurochenodeoxycholic ac- ous soluble non-starch polysaccharide are selected id, taurodeoxycholic acid, glycoursodeoxycholic ac- from the group consisting of dextran, guar gum, pec- id, taurocholic acid, glycocholic acid, their deriva- tin, indigestible soluble fiber. tives at a hydroxyl or carboxylic acid group on the 5 steroid nucleus, their salts, or their conjugates with 15. A method of ameliorating or treating at least one amines. symptomof a motor neurondisease in a subject comprising: 24. A method according to claim 15, wherein the aqueous soluble starch conversion product is select- administering to a subject a clear aqueous so- 10 ed from the group consisting of maltodextrin, dextrin, lution comprising: liquid glucose, corn syrup solid, and soluble starch.

(a) a first material selected from the group 25. A method according to claim 15, wherein the se- consisting of a bile acid, an aqueous soluble lected pH range is between approximately 1 and ap- derivative of a bile acid, a bile acid salt, and 15 proximately 10 inclusive. a bile acid conjugated with an amine by an amide linkage; 26. A method according to claim 15, wherein the first (b) a carbohydrate selected from the group material is ursodeoxycholic acid or sodium salt of consisting of an aqueous soluble starch ursodeoxycholic acid. conversion product or an aqueous soluble 20 non-starch polysaccharide; 27. A method according to claim 15, wherein the (c) water, wherein the first material and the aqueous soluble starch conversion product is mal- carbohydrate both remain in solution for all todextrin. pH values of the solution within a selected range of pH values. 25 28. A method according to claim 15, wherein the aqueous soluble non-starch polysaccharide are se- 16. A method according to claim 15, wherein the mo- lected from the group consisting of dextran, guar tor neuron disease is selected from the group con- gum, pectin, indigestible soluble fiber. sisting of amyotrophic lateral sclerosis, progressive bulbar palsy, pseudobulbar palsy, primary lateral 30 29. A method for ameliorating or treating at least one sclerosis, progressive muscular atrophy, and post- symptom of a neurodegenerative disease or at least polio syndrome. one symptom of a motor neuron disease in a subject comprising: 17. A method according to claim 16, wherein the motor neuron disease is amyotrophic lateral sclerosis. 35 administering a pharmaceutical compound to the subject; and 18. A method according to claim 17, wherein the administering to the subject a clear aqueous so- amyotrophic lateral sclerosis is advanced amyo- lution comprising: trophic lateral sclerosis. 40 (a) a first material selected from the group 19. A method according to claim 15, wherein the consisting of a bile acid, an aqueous soluble symptom is selected from the group consisting of derivative of a bile acid, a bile acid salt, and shortened lifespan and paralysis. a bile acid conjugated with an amine by an amide linkage; 20. A method according to claim 15, wherein subject 45 (b) a carbohydrate selected from the group is a mammal. consisting of an aqueous soluble starch conversion product or an aqueous soluble 21. A method according to claim 15, wherein subject non-starch polysaccharide; is a human. (c) water, wherein the first material and the 50 carbohydrate both remain in solution for all 22. A method according to claim 15, wherein the first pH values of the solution within a selected material is present in therapeutically active amount. range of pH values;

23. A method according to claim 15, wherein the first 30. A method according to claim 29, wherein the sub- material is selected from the group consisting of55 jects is a mammal. chenodeoxycholic acid, cholic acid, hyodeoxycholic acid, deoxycholic acid, 7-oxolithocholic acid, litho- 31. A method according to claim 29, wherein the sub- cholic acid, iododeoxycholic acid, iocholic acid, tau- jects is a human.

15 27 EP 2 255 812 A1 28

32. A method according to claim 29, wherein the administering a clear aqueous solution and the admin- ministering a pharmaceutical compound and the ad- istering a pharmaceutical compound occur concur- ministering a clear aqueous solution occur at the rently for each dose. same time. 5 44. A method according to claim 43, wherein the 33. A method according to claim 29, wherein the ad- clear aqueous solution further comprises the phar- ministering a pharmaceutical compound and the ad- maceutical compound. ministering a clear aqueous solution occur at different moments within a defined period of time. 45. A method according to claim 29, wherein the first 10 material is present in therapeutically active amount. 34. A method according to claim 33, wherein the administering a pharmaceutical compound occurs be- 46. A method according to claim 29, wherein the first fore the administering a clear aqueous solution. material is selected from the group consisting of chenodeoxycholic acid, cholic acid, hyodeoxycholic 35. A method according to claim 33, wherein the ad- 15 acid, deoxycholic acid, 7-oxolithocholic acid, litho- ministering a pharmaceutical compound occurs after cholic acid, iododeoxycholic acid, iocholic acid, tau- the administering a clear aqueous solution. roursodeoxycholic acid, taurochenodeoxycholic acid, taurodeoxycholic acid, glycoursodeoxycholic ac- 36. A method according to claim 29, wherein the ad- id, taurocholic acid, glycocholic acid, their deriva- ministering a pharmaceutical compound further20 tives at a hydroxyl or carboxylic acid group on the comprises administering a plurality of doses of the steroid nucleus, their salts, or their conjugates with pharmaceutical compound. amines.

37. A method according to claim 29, wherein the ad- 47. A method according to claim 29, wherein the ministering a clear aqueous solution further compris- 25 aqueous soluble starch conversion product is select- es administering a plurality of doses of the clear ed from the group consisting of maltodextrin, dextrin, aqueous solution. liquid glucose, corn syrup solid, and soluble starch.

38. A method according to claim 37, wherein the ad- 48. A method according to claim 29, wherein the se- ministering a pharmaceutical compound further30 lected pH range is between approximately 1 and ap- comprises administering a plurality of doses of the proximately 10 inclusive. pharmaceutical compound. 49. A method according to claim 29, wherein the first 39. A method according to claim 38, wherein the ad- material is ursodeoxycholic acid or sodium salt of ministering a pharmaceutical compound further35 ursodeoxycholic acid. comprises administering at least one dose of the pharmaceutical compound after the administering a 50. A method according to claim 29, wherein the clear aqueous solution. aqueous soluble starch conversion product is maltodextrin. 40. A method according to claim 38, wherein the ad- 40 ministering a pharmaceutical compound further 51. A method according to claim 29, wherein the comprises administering at least one dose of the aqueous soluble non-starch polysaccharide are se- pharmaceutical compound before the administering lected from the group consisting of dextran, guar a clear aqueous solution. gum, pectin, indigestible soluble fiber. 45 41. A method according to claim 38, wherein the ad- 52. A method according to claim 29, wherein the neu- ministering a clear aqueous solution further compris- rodegenerative disease or motor neuron disease is es administering at least one dose of the clear aque- selected from the group consisting of Parkinson’s ous solution after the administering a pharmaceuti- disease, Huntington’s disease, Alzheimer’s disease, cal compound. 50 amyotrophic lateral sclerosis, stroke, spinal cord injury, progressive bulbar palsy, pseudobulbar palsy, 42. A method according to claim 38, wherein the ad- primary lateral sclerosis, progressive muscular atro- ministering a clear aqueous solution further compris- phy, and post-polio syndrome. es administering at least one dose of the clear aqueous solution before the administering a pharmaceu- 55 53. A method according to claim 52, wherein the neu- tical compound. rodegenerative disease or motor neuron disease is amyotrophic lateral sclerosis. 43. A method according to claim 38, wherein the ad-

16 29 EP 2 255 812 A1 30

54. A method according to claim 29, wherein the clovir,forscarnet sodium, fluorouracil, ganciclovir so- pharmaceutical compound decreases motor neuron dium, idoxuridine, interferon-_, lamivudine, nevirap- death. ine, penciclovir, ribavirin, stavudine, trifluridine, val- acyclovir•HCl, zalcitabine, zidovudine, indina- 55. A method according to claim 54, wherein the 5 vir•H2SO4, ritonavir, nelfinavir•CH3SO3H, saquina- pharmaceutical compound that decreases motor vir•CH3SO3H,d- penicillamine,chloroquine, hydrox- neuron death is selected from the group consisting ychloroquine, aurothioglucose, gold sodium thioma- of Pasiniazide, Benzthiazide, Prednisolone, Men- late, auranofin levamisole, DTC, isoprinosine, me- thol, Mebhydrolin Naphthalenesulfonate, Trichlo- thyl inosine monophosphate, muramyl dipeptide, rmethiazide, Oxytetracycline, Arcaine sulphate,10 diazoxide, hydralazine•HCl, minoxidil, dipyridamole, Erythromycin, Glutathione, Trioxsalen, NylidrinHCL, isoxsuprine•HCl, niacin, nylidrin•HCl, phentolamine, Desmethyldiazepam, Thonzylamine HCL, Valproate doxazosin•CH3SO3H, prazosin•HCl, terazocin• Na, Aminophenazone, Sulfamethizole, Droperidol, HCl, clonidine•HCl, nifedipine, molsidomine, amio- 2-Thiouracil, Kynurenic acid, Fusidic acid, Leucov- darone, acetylsalicylic acid, verapamil, diltiazem, orin Ca, Sparteine sulfate, Amygdalin, Pramoxine 15 nisoldipine, isradipine, bepridil, isosorbide•dinitrate, HCL, Furosemide, Dinitolmide, Budesonide, Flopro- pentaerythrytol•tetranitrate, nitroglycerin, cimeti- pione, Fluorometholone, anti-inflammatory), N- dine, famotidine, nizatidine, ranitidine, lansoprazole, Formylmethionylphenylalanine, Thiopental Na, Lan- omeprazole, misoprostol, sucralfate, metoclopra- soprazole, Bretylium Tosylate, Cefamandole Na, mide•HCl, erythromycin, bismuth compound, alpros- Oxybendazole, Cycloleucylglycine, Dantrolene Na, 20 tadil, albuterol, pirbuterol, terbutaline•H2SO4, sal- Tetroquinone, Piperazine, Aesculin, Ethisterone, metrol, aminophylline, dyphylline, ephedrine, ethyl- Dimethadione, Griseofulvin, Acetaminosalol, Isogu- norepinephrine, isoetharine, isoproterenol, met- vacine HCL, Putrescine DIHCL, Emetine HCL, Sul- aproterenol, n-docromil, oxy triphylline, theophylline, fanilamide, Mimosine, Acetylcholine, Pralidoxime bitolterol, fenoterol, budesonide, flunisolide, beclom- Mesylate, LysylTryptophanyl-Lysine, Hecogenin,25 ethasone•dipropionate, fluticasone•propionate, co- Prednisolone acetate, Albendazole, Hydrochlorothi- deine, codeine sulfate, codeine phosphate, dex- azide, Demeclocycline HCL, Nitrofurazone, Dicloxa- tromethorphan•HBr, triamcinolone•acetonide, mon- cillin Na, alpha-Tocopherol, Tetracycline HCL, telukast sodium, zafirlukast, zileuton, cromolyn so- Fenofibrate, Probenecid, Tretinoin, Acetaminophen, dium, ipratropium bromide, nedocromil sodium ben- Hydrastinine HCL, [- dArg-2]Kyotorphin acetate, 30 zonate, diphenhydramine•HCl, hydrocodone•bitar- NMDA, Cefmetazole Na, Ribavirin, O- Benzyl-L-Ser- tarate, methadone•HCl, morphine sulfate, acetyl- ine, Picrotoxin, Oxethazine, Sulfathiazole, Trichlo- cysteine, guaifenesin, ammonium carbonate, am- rmethine, Nabumetone, Chloramphenicol, riluzole, monium chloride, antimony potassium tartarate, ginseng and its extract, glycyrrhizin and glycyrrhizic glycerin, terpin•hydrate, colfosceril palmitate, atorv- acid, derivatives of carboquinone, coenzyme Q10, 35 astatin•calcium, cervastatin•sodium, fluvastatin•so- creatine, insulin-like growth factor-1, minocycline, dium, lovastatin, pravastatin•sodium, simvastatin, mecamserin, xaliproden, gabapentin, dextromethor- picrorrhazia kurrva, andrographis paniculata, morin- phan, talampanel, IL-1, TR-500, procysteine, brain ga oleifera, albizzia lebeck, adhata vasica, curcuma derived neurotrophic factor, baclofen, tizanidin, ben- longa, momordica charantia, gymnema sylvestre, zodiazepines, glycopyrrolate, atropine, quinine,40 terminalia arjuna, azadirachta indica, tinosporia cor- phenytoin and morphine. difolia, metronidazole, amphotericin B, clotrimazole, fluconazole, haloprogin, ketoconazole, griseofulvin, 56. A method according to claim 29, wherein the itraconazole, terbinafin•HCl, econazole•HNO3, mi- pharmaceutical compound is selected from the conazole, nystatin, oxiconazole•HNO3, sulcona- group consisting of hormones, hormone antago-45 zole•HNO3, cetirizine•2HCl, dexamethasone, hy- nists, analgesic, antipyretics, anti-inflammatory drocortisone, prednisolone, cortisone, catechin and drugs, immunoactive drugs, antineoplastic drugs, its derivatives, glycyrrhizin, glycyrrhizic acid, betam- antibiotics, anti-inflammatory agents, sympathomi- ethasone, ludrocortisone•acetate, flunisolide, fluti- metic drugs, anti-infective drugs, anti- tumor agents, casone•propionate, methyl prednisolone, somato- and anesthetics. 50 statin, lispro, glucagon, proinsulin, insoluble insulins, acarbose, chlorpropamide, glipizide, glyburide, met- 57. A method according to claim 29, wherein the formin•HCl, repaglinide, tolbutamide, amino acid, pharmaceutical compound is selected from the colchicine, sulfinpyrazone, allopurinol, piroxicam, group consisting of insulin, heparin, calcitonin, amp- tolmetin sodium, indomethacin, ibuprofen, diflunisal, icillin, octreotide, sildenafil citrate, calcitriol, dihy-55 mefenamic acid, naproxen, and trientine. drotachysterol, ampomorphine, yohimbin, trazodone, acyclovir, amantadine •HCl, rimantadine•HCl, 58. A clear aqueous solution comprising: cidofovir, delavirdine•mesylate, didanosine, famci-

17 31 EP 2 255 812 A1 32

(a) a first material selected from the group con- 67. A clear aqueous solution according to claim 58, sisting of a bile acid, an aqueous soluble deriv- wherein the aqueous soluble non-starch polysac- ative of a bile acid, a bile acid salt, and a bile charide is selected from the group consisting of dex- acid conjugated with an amine by an amide link- tran, guar gum, pectin, indigestible soluble fiber. age; 5 (b) a carbohydrate selected from the group con- 68. A clear aqueous solution according to claim 58, sisting of an aqueous soluble starch conversion wherein the pharmaceutical compound is selected product or an aqueous soluble starch non- from the group consisting of Pasiniazide, Benzthi- polysaccharide; azide, Prednisolone, Menthol, Mebhydrolin, Naph- (c) a pharmaceutically effective amount of a10 thalenesulfonate, Trichlormethiazide, Oxytetracy- pharmaceutical compound that decreases mo- cline, Arcaine sulphate, Erythromycin, Glutathione, tor neuron death; and Trioxsalen, NylidrinHCL, Desmethyldiazepam, (d) water, Thonzylamine HCL, Valproate Na, Aminophena- wherein the first material, the carbohydrate, and zone, Sulfamethizole, Droperidol, 2-Thiouracil, the riluzole all remain in solution for all pH values 15 Kynurenic acid, Fusidic acid, Leucovorin Ca, of the solution within a selected range of pH val- Sparteine sulfate, Amygdalin, Pramoxine HCL, ues. Furosemide,

59. A clear aqueous solution according to claim 58, [0086] Dinitolmide, Budesonide, Flopropione, Fluor- wherein the first material is present in a neuropro- 20 ometholone, anti-inflammatory), N-Formylmethionyl- tective amount. phenylalanine, Thiopental Na, Lansoprazole, Bretylium Tosylate, Cefamandole Na, Oxybendazole, Cycloleu- 60. A clear aqueous solution according to claim 58, cylglycine, Dantrolene Na, Tetroquinone, Piperazine, wherein the pharmaceutical compound remains in Aesculin, Ethisterone, Dimethadione, Griseofulvin, solution for all pH values within the selected range. 25 Acetaminosalol, Isoguvacine HCL, Putrescine DIHCL, Emetine HCL, Sulfanilamide, Mimosine, Acetylcholine, 61. A clear aqueous solution according to claim 58, Pralidoxime Mesylate, LysylTryptophanyl-Lysine, Heco- wherein the first material is ursodeoxycholic acid. genin, Prednisolone acetate, Albendazole, Hydrochloro- thiazide, Demeclocycline HCL, Nitrofurazone, Dicloxa- 62. A clear aqueous solution according to claim 58, 30 cillin Na, alpha-Tocopherol, Tetracycline HCL, Fenofi- wherein the first material is the sodium salt of urso- brate, Probenecid, Tretinoin, Acetaminophen, Hydrasti- deoxycholic acid. nineHCL, d [-Arg-2]Kyotorphinacetate, NMDA, Cefmeta- zole Na, Ribavirin, O-Benzyl-L-Serine, Picrotoxin, Oxet- 63. A clear aqueous solution according to claim 58, hazine, Sulfathiazole, Trichlormethine, Nabumetone, wherein the first material is selected from the group 35 Chloramphenicol, riluzole, ginseng and its extract, gly- consisting of chenodeoxycholic acid, cholic acid, hy- cyrrhizin and glycyrrhizic acid, derivatives of carboqui- odeoxycholic acid, deoxycholic acid, 7- oxolithochol- none, coenzyme Q10, creatine, insulin- like growth factor- ic acid, lithocholic acid, iododeoxycholic acid, iochol- 1, minocycline, mecamserin, xaliproden, gabapentin, ic acid, tauroursodeoxycholic acid, taurochenodeox- dextromethorphan, talampanel, 1, IL- TR-500, pro- ycholic acid, taurodeoxycholic acid, glycoursodeox- 40 cysteine, brain derived neurotrophic factor, baclofen, ycholic acid, taurocholic acid, glycocholic acid, their tizanidin, benzodiazepines, glycopyrrolate, atropine, qui- derivatives at a hydroxyl or carboxylic acid group on nine, phenytoin and morphine. the steroid nucleus, their salts, or their conjugates with amines. 45 Claims 64. A clear aqueous solution according to claim 58, wherein the selected pH range is between approxi- 1. A composition comprising a clear aqueous solution mately 1 and approximately 10 inclusive. comprising:

65. A clear aqueous solution according to claim 58, 50 (a) a first material selected from soluble ursode- wherein the aqueous soluble starch conversion oxycholic acid or a sodium salt of tauroursode- product is selected from the group consisting of mal- oxycholic acid (TUDCA); todextrin, dextrin, liquid glucose, corn syrup solid, (b) a carbohydrate selected from the group con- and soluble starch. sisting of an aqueous soluble starch conversion 55 product or an aqueous soluble starch non- 66. A clear aqueous solution according to claim 58, polysaccharide; wherein the aqueous soluble starch conversion (c) water, wherein the first material and the car- product is maltodextrin. bohydrate both remain in solution for any subset

18 33 EP 2 255 812 A1 34

of the range of pH values obtainable in an aque- 13. The composition according to one of the preceding ous system sufficient for a pharmaceutical for- claims further comprising: mulation to remain in solution from preparation, to administration, to absorption in the body; (d) a pharmaceutically effective amount of a for use in the treatment of at least one symptom 5 pharmaceutical compound selected from one of of a neurodegenerative disease or at least one the following: symptom of a motor neuron disease in a subject. the pharmaceutical compound is selected 2. The composition according to claim 1, wherein the from the group consisting of Pasiniazide, neurodegenerative disease is selected from the10 Benzthiazide, Prednisolone, Menthol, Me- group consisting of Parkinson’s disease, Hunting- bhydrolin, Naphthalenesulfonate, Trichlo- ton’s disease, Alzheimer’s disease, amyotrophic lat- rmethiazide, Oxytetracycline, Arcaine sul- eral sclerosis, stroke, and spinal cord injury or the phate, Erythromycin, Glutathione, Triox- motor neuron disease is selected from the group salen, NylidrinHCL, Desmethyldiazepam, consisting of amyotrophic lateral sclerosis, progres- 15 Thonzylamine HCL, Valproate Na, Ami- sive bulbar palsy, pseudobulbar palsy, primary lat- nophenazone, Sulfamethizole, Droperidol, eral sclerosis, progressive muscular atrophy, and 2-Thiouracil, Kynurenic acid, Fusidic acid, post-polio syndrome. Leucovorin Ca, Sparteine sulfate, Amygda- lin, Pramoxine HCL, Furosemide, Dinitol- 3. The composition according to claim 1, wherein the 20 mide, Budesonide, Flopropione, Fluor- neurodegenerative disease or the motor neuron dis- ometholone, antiinflammatory), N-Formyl- ease is amyotrophic lateral sclerosis. methionylphenylalanine, Thiopental Na, Lansoprazole, Bretylium Tosylate, Cefa- 4. The composition according to claim 1 or claim 3, mandole Na, Oxybendazole, Cycloleucylg- wherein the amyotrophic lateral sclerosis is ad-25 lycine, Dantrolene Na, Tetroquinone, Pip- vanced amyotrophic lateral sclerosis. erazine, Aesculin, Ethisterone, Dimethadi- one, Griseofulvin, Acetaminosalol, Isogu- 5. The composition according to claim 1, wherein the vacine HCL, Putrescine DIHCL, Emetine symptom is selected from the group consisting of HCL, Sulfanilamide, Mimosine, Acetylcho- shortened lifespan and paralysis. 30 line, Pralidoxime Mesylate, LysylTryptoph- anyl-Lysine, Hecogenin, Prednisolone ace- 6. The composition according to claim 1, wherein the tate, Albendazole, Hydrochlorothiazide, subject is a mammal. Demeclocycline HCL, Nitrofurazone, Di- cloxacillin Na, alpha-Tocopherol, Tetracy- 7. The composition according to claim 1, wherein the 35 cline HCL, Fenofibrate, Probenecid, Tretin- subject is a human. oin, Acetaminophen, Hydrastinine HCL, d[- Arg-2]Kyotorphin acetate, NMDA, Ce- 8. The composition according to claim 1, wherein the fmetazole Na, Ribavirin, Benzyl-L-Serine, first material is present in therapeutically active Picrotoxin, Oxethazine, Sulfathiazole, amount. 40 Trichlormethine, Nabumetone, Chloram- phenicol, riluzole, ginseng and its extract, 9. The composition according to claim 1, wherein the glycyrrhizin and glycyrrhizic acid, deriva- aqueous soluble starch conversion product is select- tives of carboquinone, coenzyme Q10, cre- ed from the group consisting of maltodextrin, dextrin, atine, insulin-like growth factor-I, minocy- liquid glucose, corn syrup solid, and soluble starch. 45 cline, mecamserin, xaliproden, gabapentin, dextromethorphan, talampanel, 1, IL- 10. The composition according to claim 1, wherein the TR500, procysteine, brain derived neuro- selected pH range is between approximately 1 and trophic factor, baclofen, tizanidin, benzodi- approximately 10 inclusive. azepines, glycopyrrolate, atropine, quinine, 50 phenytoin and morphine 11. The composition according to claim 9, wherein the that decreases motor neuron death, and aqueous soluble starch conversion product is mal- wherein the first material, the carbohydrate, todextrin. and the pharmaceutically effective amount of the pharmaceutical compound all remain 12. The composition according to claim 1, wherein the 55 in solution for all pH values of the solution aqueous soluble non-starch polysaccharide are se- within a selected range of pH values. lected from the group consisting of dextran, guar gum, pectin, indigestible soluble fiber. 14. The composition according to claim 13, wherein the

19 35 EP 2 255 812 A1 36

first material is present in a neuroprotective amount.

15. A composition comprising a clear aqueous solution comprising 5 (a) a soluble tauroursodeoxycholic acid (TUD- CA); (b) water, whereintauroursodeoxycholic acid remain in solution for any subset of the range of pH values obtainable in an aqueous system suf- 10 ficient for a pharmaceutical formulation to remain in solution from preparation, to administration, to absorption in the body;

15

20

25

30

35

40

45

50

55

20 EP 2 255 812 A1

21 EP 2 255 812 A1

22 EP 2 255 812 A1

23 EP 2 255 812 A1

24 EP 2 255 812 A1

25 EP 2 255 812 A1

26 EP 2 255 812 A1

27 EP 2 255 812 A1

28

Web Analytics