(19) TZZ ¥ _T

(11) EP 2 329 829 B1

(12) EUROPEAN PATENT SPECIFICATION

(45) Date of publication and mention (51) Int Cl.: of the grant of the patent: A61K 31/7072 (2006.01) A61K 31/14 (2006.01) 16.04.2014 Bulletin 2014/16 A61K 31/685 (2006.01) A61P 25/28 (2006.01)

(21) Application number: 10075661.8

(22) Date of filing: 30.07.1999

(54) Use of in combination with choline for the treatment of neurological disorders Verwendung von Uridin in Kombination mit Cholin zur Behandlung neurologischer Erkrankungen Utilisation de l’uridine en combinaison avec la choline pour le traitement des maladies neurologiques

(84) Designated Contracting States: • CACABELOSR ET AL: "THERAPEUTIC EFFECTS AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU OF CDP-CHOLINE IN ALZHEIMER’S DISEASE MC NL PT SE COGNITION, BRAIN MAPPING, CEREBROVASCULAR HEMODYNAMICS, AND (30) Priority: 31.07.1998 US 95002 P IMMUNE FACTORS", ANNALS OF THE NEW YORK ACADEMY OF SCIENCES, NEW YORK (43) Date of publication of application: ACADEMY OF SCIENCES, NEW YORK, NY, US, 1 08.06.2011 Bulletin 2011/23 January 1996 (1996-01-01), pages 399-403, XP008065562, ISSN: 0077-8923 (62) Document number(s) of the earlier application(s) in • SPIERS P A ET AL: "CITICOLINE IMPROVES accordance with Art. 76 EPC: VERBAL MEMORY IN AGING", ARCHIVES OF 09173495.4 / 2 145 627 NEUROLOGY, AMERICAN MEDICAL 07116909.8 / 1 870 103 ASSOCIATION, CHICAGO, IL, US, vol. 53, no. 5, 99937631.2 / 1 140 104 1 May 1996 (1996-05-01), pages 441-448, XP008028412, ISSN: 0003-9942 (73) Proprietor: Massachusetts Institute of Technology • WEISS G B: "Metabolism and actions of CDP- Cambridge, Massachusetts 02142-1601 (US) choline as an endogenous compound and administered exogenously as citicoline.", LIFE (72) Inventors: SCIENCES 1995 LNKD- PUBMED:7869846, vol. • Watkins, Carol 56, no. 9, 1995, pages 637-660, XP002640081, Cambridge, MA 02142 (US) ISSN: 0024-3205 • Wurtman, Richard J. • D’ORLANDO K J ET AL: "Citicoline (CDP- Boston, MA 02116 (US) choline): mechanisms of action and effects in ischemic brain injury.", NEUROLOGICAL (74) Representative: Pearl Cohen Zedek Latzer Baratz RESEARCH AUG 1995 LNKD- PUBMED: 7477743, UK LLP vol. 17, no. 4, August 1995 (1995-08), pages 15 Old Bailey 281-284, XP009149054, ISSN: 0161-6412 London EC4M 7EF (GB) • PICCOLI F ET AL: "CDP- choline in the treatment of chronic cerebrovasculopathies", ARCHIVES (56) References cited: OF GERONTOLOGY AND GERIATRICS, WO-A-97/45127 DE-A1- 2 508 474 ELSEVIER, AMSTERDAM, NL, vol. 18, no. 3, 1 May DE-A1- 2 629 845 US-A- 4 221 784 1994 (1994-05-01), pages 161-168, XP023512323, US-A- 4 609 647 US-A- 4 960 759 ISSN: 0167-4943, DOI: DOI: 10.1016/0167-4943(94) US-A- 5 567 689 90010-8 [retrieved on 1994-05-01]

Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). EP 2 329 829 B1

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• KARKISHCHENKO N N; MAKLYAKOV YU S; STRADOMSKII B V: "Use of uridine as antidepressant - shows its reduced toxicity and elimination of several harmful side effects", DERWENT, 30 November 1993 (1993-11-30), XP002302971, • DATABASE BIOSIS [Online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 1986, MERLINI G ET AL: "EFFECTS OF LARGE DOSES OF AND URIDINE IN ELDERLY PATIENTS WITH NEUROPSYCHOLOGICAL DISTURBANCES CAUSED BY VASCULAR AND CEREBRAL METABOLIC INSUFFICIENCY", XP002640124, Database accession no. PREV198783027367 & GAZZETTA MEDICA ITALIANAARCHIVIO PER LE SCIENZE MEDICHE, vol. 145, no. 6, 1986, pages 379-390, ISSN: 0393-3660

2 1 EP 2 329 829 B1 2

Description ible. Moreover, Dawson himself states that he was una- ble to recover a with spectrophotometric char- [0001] The present invention relates to uridine or a uri- acteristics of cytidine and admits that his conclusions dine source in combination with choline, a choline pre- were based on probabilistic guessing. Thus, the alleged cursor, a choline salt or ester, or a mixture thereof, for 5 phenomenon observed by Dawson may have been due use in treating neurological disorders associated with the to misinterpretation of an experimental artifact as it is dopaminergic pathway, such as schizophrenia or Parkin- now known that experimentally measurable cytidine can son’s disease, cognitive dysfunction stroke andbe easily confused with tyrosine, which is chemically un- ischemia. related amino acid compound (see Fig. 1). 10 [0005] Thus, even though an catalyzing the DESCRIPTION OF THE RELATED ART conversion of uridine to cytidine may exist in rats its ac- tivity is not sufficiently potent to raise the levels of cytidine [0002] This invention stems from unexpected discov- to a level that can be measured and ascertained beyond ery that increase in levels of uridine following the admin- any doubt. Thus, these levels may be not sufficient to istration of uridine or uridine source to certain animals 15 warrant practical exploitation for clinical application. In- comprising human patients, leads to increased levels of deed nowhere in Dawson publication is there a sugges- cytidine in a human body and particularly in the human tion or an attempt to make a suggestion that the uridine brain. Thus, administering uridine or uridine precursors to cytidine conversion process can be useful for any med- to human patients in need thereof can be as beneficial ical modality. In addition, as it is the case with many other as administration of cytidine or cytidine precursors. How- 20 and metabolic pathways, this particular en- ever, the potential benefit of uridine or uridine source zyme may have been present in rats but not in humans. administration is overwhelmingly greater than the benefit One skilled in related art knows that a discovery of a of cytidine administration. This is due to the fact that cy- biological process in one species of an animal, e.g., rat, tidine, as opposed to uridine, either cannot cross or is does not necessarily means that a similar process is much less efficient than uridine in crossing the blood- 25 present in another animal, e.g., man. Based on that one brain barrier (Cornford et al., independent blood-brain skilled in the art will be not sufficiently motivated to exploit barrier transport systems for nucleic acid precursors. Bi- this phenomenon for any useful purposes other than an ochim. Biophys. Acta 349:211-219, 1975). experimental tool to study enzyme metabolism in rats. [0003] According to the knowledge relating to the me- Consequently, the prior art is silent in regard to the use tabolism of pyrimidine compounds, enzymes are known 30 of the process of uridine to cytidine conversion for any in the art, such as cytidine deaminase (EC 3.5.4.5), which meaningful application. converts cytidine into uridine. Cytidine deaminase can [0006] Uridine is a pyrimidine and is essen- be found in some prokaryotes and eukaryotes including tial in the synthesis of ribonucleic acids and tissue gly- humans, primates, and some rodents although some cogens such as UDP glucose and UTP glucose. Medical species lack this enzyme. However, according to EC (en- 35 uses of uridine alone are limited to treatment of genetic zyme classification) list there are no known examples of disorders related to deficiencies of pyrimidine synthesis aminase-like enzymes, which are capable of opposite such as orotic aciduria (Becroft DM, et al., Hereditary action, i.e., converting uridine into cytidine. orotic aciduria: longterm therapy with uridine and a trial [0004] The prior art relating to the process of uridine of . J Pediatr. 1969 Nov; 75 (5): 885-891). Other to cytidine conversion is also limited. Only one publica- 40 less common uses of uridine alone are known such as tion, citing two earlier references, seems to exist, wherein treatment of seizures and epilepsy (Roberts CA, et al., it was suggested that a soluble fraction of the rat liver Uridine anticonvulsant effects: selective control of nucl- and possibly of the brain may catalyze in vitro and in vivo eoside incorporation in experimental epilepsy. Epilepsia. the conversionof uridine nucleotide to cytidine nucleotide 1974 Dec; 15(4): 479-500). Most commonly, uridine is (Dawson. Enzymic conversion of uridine nucleotide to 45 used in combination with cytidine (Monticone GF, et al., cytidine precursor by rat brain. J. Neurochem. 15:31-34, On the therapeutic use of the nucleosides, cytidine and 1968). Even though this report implicated the possibility uridine, in some neurological diseases. Minerva Med. of such an enzyme reaction in rats the activity of the en- 1966 Dec 19; 57 (101): 4348-4352). The uses of this zyme does not appear to be sufficiently potent. As com- particular dual combination range from liver and kidney pared to the initial, administered dose of uridine (consid- 50 diseases to a number of neurological and cerebrovascu- ered as 100%), the highest levels of newly converted lar diseases but such uses are irrelevant to the present cytidine in vivo were 12.4% in the liver and 9% in the invention directed at the use of uridine without concom- brain. The conversion rates in vitro were 5.4% in the liver itant use with cytidine. and 8.05% in the brain. Thus, maximum observed levels [0007] U.S. Pat. No. 4,960,759, issued to De Luca et were within 5.4-12.4 % range. From a statistical point of 55 al., on October 2, 1990 discloses the pharmacological view all these figures are within the range of a typical use of uridine in the treatment of nervous disorders such scatter in a gamma counter (15%) and practitioner in the as schizophrenia and Parkinson’s disease. De Luca et art can dismiss them either as insignificant or irreproduc- al., teach that the benefit of uridine is due to increase in

3 3 EP 2 329 829 B1 4 cholecystokinin levels in the brain, which in turn improves sible. It is also possible to imagine the methods of treating dopamine functioning and results in therapeutic benefit. neurological diseases associated with dopaminergic Said benefit is described as a reduction in symptoms of pathway, e.g., schizophrenia and Parkinson’s disease Parkinson’s disease, which are tremor and rigidity. As as treated by combination therapy in which uridine is one the preferred embodiment of the instant invention is treat- 5 of constituents. ment of neurological disorders unrelated to schizophre- [0011] Thus, none of the prior art patents or references nia and Parkinson’s disease it is clear that the teachings have anticipated or made the instant invention obvious. by De Luca et al., are irrelevant to this invention. The present invention is thus unique and stands out in [0008] The U.S. Pat. No. 5,470, 838, issued to von the light of the prior art. Borstel et al., on November 28, 1995 discloses the meth- 10 od of delivering exogenous uridine or cytidine in form of SUMMARY OF THE INVENTION acylated uridine or cytidine and said compounds as use- ful in treating cardiac insufficiency, myocardial infarction, [0012] This invention is based upon unexpected dis- and cirrhosis of the liver. Von Borstel et al., propose to covery that uridine administration in humans leads to in- use both forms of since it was not obvious 15 crease in systemic and brain cytidine. Thus the present to them that uridine alone is effective. The absolute re- invention relates to a preparation for use in the treatment quirement of both cytidine and uridine was due to the ofa neurologicaldisorder selected from neurologicaldys- lack of knowledge and anticipation in the prior art that order associated with the dopaminergic pathway, cogni- uridine might convert into cytidine, especially in humans. tive dysfunction, stroke or ischemia the preparation com- One skilled in the art will recognize that the disclosed20 prising (a) uridine or a uridine source and (b) choline, a composition matter is different and diseases to be treated choline precursor, a choline salt or ester, or a mixture are not the same as in the present invention. thereof. The neurological disorder may be cognitive dys- [0009] The U.S. Pat. Nos. 5,141,943; 5,567,689; and function, schizophrenia, Parkinson’s disease, stroke or 5,723,449 disclose various methods and compositions ischemia. The choline precursor may be sphingomyelin, to raise levels of uridine in the blood as useful for reducing 25 cytidine-diphospho-choline, acylglycerophosphocholine toxicity of pyrimidine nucleoside drugs such as AZT and (AGP-choline), phosphatidylcholine (PC), or a combina- 5-Fluouracil for AIDS and cancer therapy respectively. It tion thereof. The choline salt may be choline chloride, is apparent to anyone skilled in the art that these teach- choline bitartrate, choline stearate or mixtures thereof. ings have nothing in common with the present invention. The uridine or uridine source may be administered in [0010] Although all of these patents and prior art ref- 30 dosages between about 10 mg and 10 grams per day. erences disclose at least one or another aspect of the The uridine may be a uridine salt or a food product con- instant invention none of them taught specifically that cy- taining uridine. tidine levels can be raised in humans by administering [0013] The terms" uridine precursor" or " uridine uridine or uridine source as useful for the treatment of source" or" uridine prodrug" are used interchangeably certain neurological or brain disorders. These disorders 35 and as defined hereinafter mean compounds, e.g., urid- comprise disorders associated with aging such as mem- ine salts or food products containing uridine, that trans- ory decline and age related decline in cognition functions. form into uridine upon administration to a host such as These disorders also comprise memory decline and re- human. lated cognition dysfunction associated with pathological [0014] Choline is involved in metabolism and lipid conditions like Alzheimer’s disease, Pick’s disease, Lewy 40 transport and is a component of a number of important Body disease, and/or dementias like Huntington’s dis- biological compounds including the membrane phos- ease and AIDS dementia. Other cognitive dysfunctions, pholipids like lecithin and sphingomyelin. Choline is also i.e., disorders of attention, alertness, concentration, fo- a precursor of acetylcholine - one of most important neu- cus, and dyslexia can also be treated. Other uses of uri- rotransmitters. Although a required nutrient for several dine therapy can be imagined such as treatment of mood 45 animal species, choline is not currently designated as and emotional disorders, e.g., mania, depression, stress, essential for humans. However, recent clinical studies panic, anxiety, insomnia, dysthemia, psychosis, season- show it to be essential for normal liver function. Addition- al effective disorders and bipolar disorders. Neurological ally, a large body of evidence from the fields of molecular diseases like ataxias, including Friedreich’s ataxia and and cell biology shows that certain phospholipids play a movement disorders like tardive dyskinesia can also be 50 critical role in generating second messengers for cell treated. Method of treating stroke, cerebral thrombosis, membrane signal transduction. This process involves a ischemia, and related cerebrovascular diseases result- cascade of reactions that translate an external cell stim- ing from hypoxia as well as behavioral and neurological ulus such as a hormone or growth factor into a change syndromes seen after brain trauma, spinal cord injury in cell transport, metabolism, growth, function, or gene and/or anoxia can be also imagined. Methods of treating 55 expression. Disruptions in phospholipid metabolism can diseases of the peripheral nervous system, e.g., neu- interfere with this process and may underlie certain dis- romuscular disorders like myasthenia gravis, the post- ease states such as cancer and Alzheimer’s disease. polio syndrome, and muscular dystrophies are also pos- However, choline alone is not useful as a therapeutical

4 5 EP 2 329 829 B1 6 modality. In light of the instant invention choline or choline dopaminergic pathway, e.g., schizophrenia and Parkin- precursors are appropriate in considering them in com- son’s disease as treated by uridine alone are specifically bination with uridine or uridine source. excluded. [0015] It is thus a further object of this invention to es- [0019] The following examples illustrate the invention tablish a synergy between uridine and various com-5 and are not intended to be limiting unless otherwise spec- pounds affecting cholinergic pathway and/or phospholi- ified. pid metabolism. Among them are CDP-choline, choline, choline salts, lecithin or phosphatidylcholine. EXAMPLE 1.

BRIEF DESCRIPTION OF THE DRAWINGS 10 [0020] In this example a method is established that overcomes the problem of the coincidence of cytidine [0016] and tyrosine peaks when tested by a standard HPLC method for measuring various nucleosides in biological Fig. 1 illustrates the coincidence of cytidine and ty- fluids (see Fig. 1). By using the standard HPLC method, rosinepeaks (6. 59) whentested bya standard HPLC 15 one can, however, easily distinguish the uridine peak method. from the cytidine peak. Detailed description of the HPLC Fig. 2 illustrates distinct cytidine (3. 25) and tyrosine method can be found for example in Lopez-Coviella et (2. 92) peaks when tested by a modified HPLC meth- al., (Evidence that 5’-cytidinephosphocholine can affect od, which utilizes elution buffer with low methanol. brain phospholipid composition by increasing choline Fig. 3 shows the ratio of uridine (100%) to cytidine 20 and cytidine plasma levels. J. Neurochemistry 65: in plasma after oral administration of 250 milligram 889-894, 1995). Modified HPLC is carried out in a same per kg of body weight (mg/kg) of uridine. way as standard HPLC except that elution buffer contains Fig. 4 shows the ratio of uridine (100%) to cytidine low amount of methanol (0.1%) instead of formic acid in the brain after oral administration of 250 milligram and as a result cytidine can be distinguished from unre- per kg of body weight (mg/kg) of uridine. 25 lated compound tyrosine (Fig. 2). This method is useful in distinguishing cytidine from masking effect of amino DESCRIPTION OF THE PREFFERED EMBODIMENTS acid tyrosine, which may concomitantly be present in tested biological fluid, e.g., plasma or cerebrospinal fluid [0017] The compositions prepared according to the (CSF). Due to the overlap between cytidine and tyrosine present invention increase systemic and brain levels of 30 it is very likely that the results of all the prior art studies cytidine in a human patient via administration of uridine dealing with cytidine measurement, including present in- or a uridine source in combination with choline, a choline ventors’ own studies supra, were interpreted incorrectly. precursor, a choline salt, a choline ester, and thus are for use in treating neurological disorders associated with EXAMPLE 2. the dopaminergic pathway such as schizophrenia or Par- 35 kinson’s disease, cognitive dysfunction, stroke and [0021] Gerbils rather than rats or other rodents are se- ischemia. The compositions optionally additionally en- lected for this example, as the pyrimidine metabolism of compass drugs that increase uridine availability. Among said gerbils is closer to humans. For practical and ethical such drugs are drugs that act as uridine phosphorylase reasons humans cannot always be used for certain ex- inhibitors like benzyl or derivatives thereof. 40 perimental studies and those skilled in the art generally Among such drugs are also drugs that act as uridine se- recognize that the gerbil model is equivalent to a human cretion inhibiting compounds like dilazep or hexobend- model. Indeed, gerbils are the choice model for certain ine. Among such drugs are drugs that act as uridine renal human diseases and brain disorders such as cerebral transport competitors like L-uridine, L-2’, 3’-dideoxyurid- ischemia (Ginsburg et al., Rodent models of cerebral ine, and D-2’, 3’-dideoxyuridine. The disclosed compo- 45 ischemia. Stroke 20: 1627-1642, 1989). Gerbils are given sitions are beneficial to a human patient in need thereof orally uridine and 60 minutes later plasma and brain lev- and act in synergy with uridine in generation of phosphol- els of cytidine and uridine are measured by modified ipids involved in brain cell membrane formation and re- HPLC method described in Example 1. The Fig. 3 shows pair. More specifically choline-based compounds are the relative ratio between uridine and cytidine levels in contemplated as compounds acting in synergy with uri- 50 plasma after oral administration of 250 milligram per kg dine or uridine source. Among them are choline, choline of body weight (mg/kg) of uridine. The Fig. 4 shows the salts or esters, such as choline bitartrate or stearate or relative ratio between uridine and cytidine levels in the the like, or compound that dissociate to choline, such as brain after oral administration of 250 mg/kg of uridine. sphingomyelin, cytidine-diphospho-choline or citicoline These results indicate that the metabolic processing of or CDP-choline, acylglycerophosphocholines, e.g., leci- 55 uridine in the brain is different than systemic processing thin, lysolecithin, glycerophosphatidylcholine, mixtures of uridine in plasma. The results also indicate that uridine, thereof or the like. when transported into the brain, is readily converted to [0018] Neurological diseases associated withcytidine and this conversion is more efficient in the brain

5 7 EP 2 329 829 B1 8 than in plasma. Similar experiments are also carried out also doses of drugs combination, which produce desired in humans wherein instead of measuring brain levels of effect in at least 10% of treated patients population. Dos- nucleosides the CSF levels are measured. The finding es are administered either as a single dose or divided in that uridine is readily converted to cytidine especially in several doses. Drugs are administered orally such as in the brain is totally unexpected and constitutes the basis 5 tablet, capsule or liquid form or parenterally by intrave- for the present invention. nous, intramuscular or subcutaneous injection. [0028] When necessary and as required by the exigen- EXAMPLE 3 cy of the therapy uridine is administered in combination with other compounds that act either synergistically or in [0022] In Example 3 a clinical study is carried out with 10 additive manner. This reduces the therapeutic dose of the goal of treating memory disorders and cognitive dys- administered drugs, thereby reducing potential undesir- functions associated with aging as well as memory de- able side effects and frequency of drug administration. cline and cognitive dysfunction associated with patho- Compounds that act in such a manner are chemical sub- logical conditions like Alzheimer’s disease, Pick’s dis- stances participating in cholinergic metabolism. For ex- ease, Lewy Body disease, and/or dementias like Hunt- 15 ample, compounds administered along with uridine are ington’s disease and AIDS dementia. Patients with non- following choline-based compounds: choline, choline pathological dementia associated with aging are also in- salts or esters, such as choline bitartrate or stearate or cluded. Oral doses of uridine alone ranging from 5 mg to the like, or compound that dissociate to choline, such as 50,000 mg are administered daily to five male and five sphingomyelin, cytidine-diphospho-choline or citicoline female patients suffering from one of the diseases listed 20 or CDP-choline, acylglycerophosphocholines, e.g., leci- above. The adjustment in dosage to select optimally ef- thin, lysolecithin, glycerophosphatidylcholine, mixtures fective pharmaceutical dose is a routine procedure well thereof or the like. The choline or compound that disso- known to the practitioner skilled in the relevant art. The ciates into choline is administered so that a choline level terms"therapeutically" or "pharmaceutically" or "pharma- of at least about 20-30 nanomoles and usually between cologically effective dose" of a drug as used hereinafter 25 10 and 50 nanomoles is attained in patient’s blood or means the amount (dosage) of the drug that provides brain. desired clinical effect in at least 10% of treated[0029] The pharmacologically effective doses are with- patients" population. in about 20 mg and 50 g/day range, preferably between [0023] Various other uridine-based compounds other about 100 mg and 10 g/day. Doses are administered ei- than uridine itself serve as uridine source or uridine pre- 30 ther as a single dose or divided in several doses, e.g., cursors. These are uridine-rich food or dietary products 10 mg to 1 g/cap or tab. The minimal duration of the like algae; salts of uridine like uridine phosphates, acylat- therapy is at least one day but longer periods of time are ed uridine or the like. usually required according to the exigency of the therapy. [0024] If required by the exigency of the therapy, ther- If needed, the usual time period spans from one day to apeutically or pharmacologically effective doses of acyl 35 the period of lifetime. When these compounds are not derivatives of uridine or mixtures thereof like those dis- available in pure form the active ingredient comprises at closed in U.S. Pat. No. 5,470,838 are also administered. leastabout 20-30 percent of theweight of the preparation. [0025] If required by the exigency of the therapy, ther- The clinical study is continued for at least 1 day or longer apeutically or pharmacologically effective doses of urid- as required by the exigencies of the therapy. In general, ine phosphorylase inhibitors like 5-benzyl barbiturate de- 40 the dose administered, the frequency of administration rivatives or mixtures thereof as disclosed in U.S. Pat. No. and the duration of the treatment will vary as a function 5, 141, 943 are also administered. ofthe condition of the patient and isdetermined according [0026] If required by the exigency of the therapy, ther- to standard clinical procedures known to the practitioner apeutically or pharmacologically effective doses of urid- skilled in the relevant art. ine secretion inhibiting compounds like dilazep, hex-45 [0030] In Example 4 a clinical study is carried out, obendine, or mixtures thereof as disclosed in U.S. PAT. which by its design and principles is similar to clinical No. 5,567, 689 are also administered. study of Example 3 except that patients enrolled in this [0027] If required by the exigency of the therapy, ther- study are patients with cognitive dysfunction, i.e., disor- apeutically or pharmacologically effective doses of com- ders of attention, alertness, concentration, focus, and pounds which compete with uridine in kidney clearance 50 dyslexia. like L-uridine, L-2’,3’-dideoxyuridine, and D-2’,3’-dideox- [0031] In Example 5 a clinical study is carried out, yuridine or mixtures thereof as disclosed in U.S. Pat. Nos. which by its design and principles is similar to clinical 5,723,449 and 5,567,689 are also administered. Thera- study of Example 3 except that patients enrolled in this peutically or pharmacologically effective doses of uridine study are patients with strokes or ischemia. as defined herein are also doses that produce blood or 55 [0032] In Example 6 a clinical study is carried out, brain levels of cytidine ranging between 0. 1 micromole which by its design and principles is similar to clinical (pM) and 1 millimole (mM). In general, therapeutically or study of Example 3 except that patients enrolled in this pharmacologically effective doses as defined herein are study are patients with neurological diseases associated

6 9 EP 2 329 829 B1 10 with dopaminergic pathway, e.g., schizophrenia and Par- in the choline salt is choline chloride, choline kinson’s disease and said diseases are treated by com- bitartrate, choline stearate or mixtures thereof. bination therapy in which uridine is one of constituents. [0033] When possible clinical studies as disclosed in 10. The use according to any one of claims 7 to 9, where- any preceding examples are preceded by in vivo studies 5 in said uridine or uridine source is administered in in animal models, e.g., gerbil model, according to proce- dosages between about 10 mg and 10 grams per dures established in the art. day.

Claims 10 Patentansprüche

1. A preparation for use in the treatment of a neurolog- 1. Eine Zubereitung zur Verwendung in der Behand- ical disorder, said preparation comprising (a) uridine lung einer neurologischen Erkrankung, wobei die or a uridine source and (b) choline, a choline precur- Zubereitung Folgendes umfasst: (a) Uridin oder eine sor, a choline salt or ester, or a mixture thereof,15 Uridinquelle und (b) Cholin, einen Cholin-Vorläufer, wherein the neurological disorder is selected from a ein Cholinsalz oder -ester oder eine Mischung da- neurological disorder associated with the dopamin- von, wobei die neurologische Erkrankung ausge- ergic pathway, cognitive dysfunction, stroke or wählt wird aus einer mit der dopaminergen Bahn ver- ischemia. bundenen neurologischen Erkrankung, kognitiver 20 Fehlfunktion, Schlaganfall oder Ischämie. 2. The preparation for use according to claim 1, wherein the neurological disorder associated with the 2. Die Zubereitung zur Verwendung gemäß Anspruch dopaminergic pathway comprises schizophrenia 1,wobei die mit der dopaminergen Bahn verbundene and Parkinson’s disease. neurologische Erkrankung Schizophrenie und Par- 25 kinson-Krankheit umfasst. 3. The preparation for use according to any one of the preceding claims, wherein the choline precursor is 3. Die Zubereitung zur Verwendung gemäß einem der sphingomyelin, cytidine-diphospho-choline, acylg- vorhergehenden Ansprüche, wobei der Cholin-Vor- lycerophosphocholine (AGP-choline), phosphatidyl- läufer Sphingomyelin, Cytidindiphosphocholin, Acyl- choline (PC), or a combination thereof. 30 glycerophosphocholin (AGP-Cholin), Phosphatidyl- cholin (PC) oder eine Kombination davon ist. 4. The preparation for use according to any one of the preceding claims, wherein the choline salt is choline 4. Die Zubereitung zur Verwendung gemäß einem der chloride, choline bitartrate, choline stearate or mix- vorhergehenden Ansprüche, wobei das Cholinsalz tures thereof. 35 Cholinchlorid, Cholinbitartrat, Cholinstearat oder Mi- schungen davon ist. 5. The preparation for use according to any one of the preceding claims, wherein said uridine or uridine 5. Die Zubereitung zur Verwendung gemäß einem der source is administered in dosages between about vorhergehenden Ansprüche, wobei das Uridin oder 10 mg and 10 grams per day. 40 die Uridinquelle in Dosierungen zwischen etwa 10 mg und 10 g pro Tag verabreicht wird. 6. The preparation for use according to any one of the preceding claims, wherein said uridine comprises 6. Die Zubereitung zur Verwendung gemäß einem der uridine salts or a food product containing uridine. vorhergehenden Ansprüche, wobei das Uridin Uri- 45 dinsalze oder ein uridinhaltiges Nahrungsmittel um- 7. Use of a preparation comprising (a) uridine or a uri- fasst. dine source and (b) choline, a choline precursor, a choline salt or ester, or a mixture thereof in the man- 7. Verwendung einer Zubereitung, umfassend (a) Uri- ufacture of a composition for treating a neurological din oder eine Uridinquelle und (b) Cholin, einen Cho- disorder selected from cognitive dysfunction, schiz- 50 lin-Vorläufer, ein Cholinsalz oder -ester oder eine ophrenia, Parkinson’s disease, stroke or ischemia. Mischung davon bei der Herstellung einer Zusam- mensetzung zur Behandlung einer neurologischen 8. The use according to claim 7, wherein the choline Erkrankung, ausgewählt aus einer kognitiven Fehl- precursor is sphingomyelin, cytidine-diphospho- funktion, Schizophrenie, Parkinson-Krankheit, choline, acylglycerophosphocholine (AGP-choline), 55 Schlaganfall oder Ischämie. phosphatidylcholine (PC), or a combination thereof. 8. Die Verwendung gemäß Anspruch 7, wobei der Cho- 9. The use according to any one of claims 7 or 8, where- lin-Vorläufer Sphingomyelin, Cytidindiphosphocho-

7 11 EP 2 329 829 B1 12

lin, Acylglycerophosphocholin (AGP-Cholin), Phos- tion pour le traitement d’un trouble neurologique sé- phatidylcholin (PC) oder eine Kombination davon ist. lectionné parmi dysfonctionnement cognitif, schi- zophrénie, maladiede Parkinson, attaque vasculaire 9. Die Verwendung gemäß einem der Ansprüche 7 cérébrale ou ischémie. oder 8, wobei das Cholinsalz Cholinchlorid, Cholin- 5 bitartrat, Cholinstearat oder Mischungen davon ist. 8. L’utilisation selon la Revendication 7, où le précur- seur de choline est sphingomyéline, cytidine-diphos- 10. Die Verwendung gemäß einem der Ansprüche 7 bis pho-choline, acylglycérophosphocholine (AGP-cho- 9, wobei das Uridin oder die Uridinquelle in Dosie- line), phosphatidylcholine (PC), ou une combinaison rungen zwischen etwa 10 mg und 10 g pro Tag ver- 10 de ceux-ci. abreicht wird. 9. L’utilisation selon l’une quelconque des Revendica- tions 7 ou 8, où le sel de choline est chlorure de Revendications choline, bitartrate de choline, stéarate de choline, ou 15 des mélanges de ceux-ci. 1. Une préparation destinée à une utilisation dans le traitement d’un trouble neurologique, ladite prépa- 10. L’utilisation selon l’une quelconque des Revendica- ration contenant (a) uridine ou une source d’uridine tions 7 à 9, où ladite uridine ou source d’uridine est et (b) choline, un précurseur de choline, un sel ou administrée dans des dosages entre environ 10 mg ester de choline, ou un mélange de ceux-ci, où le 20 et 10 grammes par jour. trouble neurologique est sélectionné parmi un trou- ble neurologique associé à la voie dopaminergique, un dysfonctionnement cognitif, une attaque vascu- laire cérébrale ou une ischémie. 25 2. La préparation destinée à une ut ilisation selon la Re- vendication 1, où le trouble neurologique associé à la voie dopaminergique comprend schizophrénie et maladie de Parkinson. 30 3. La préparation destinée à une utilisation selon l’une quelconque des Revendications précédentes, où le précurseur de choline est sphingomyéline, cytidine- diphospho-choline, acylglycérophosphocholine (AGP-choline), phosphatidylcholine (PC), ou une 35 combinaison de ceux-ci.

4. La préparation destinée à une utilisation selon l’une quelconque des Revendications précédentes, où le sel de choline est chlorure de choline, bitartrate de 40 choline, stéarate de choline, ou des mélanges de ceux-ci.

5. La préparation destinée à une utilisation selon l’une quelconque des Revendications précédentes, où la- 45 dite uridine ou source d’uridine est administrée dans des dosages entre environ 10 mg et 10 grammes par jour.

6. La préparation destinée à une utilisation selon l’une 50 quelconque des Revendications précédentes, où la- dite uridine comprend des sels d’uridine ou un pro- duit alimentaire contenant de l’uridine.

7. L’utilisation d’une préparation contenant (a) uridine 55 ou une source d’uridine et (b) choline, un précurseur de choline, un sel ou ester de choline, ou a un mé- lange de ceux-ci dans la fabrication d’une composi-

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REFERENCES CITED IN THE DESCRIPTION

This list of references cited by the applicant is for the reader’s convenience only. It does not form part of the European patent document. Even though great care has been taken in compiling the references, errors or omissions cannot be excluded and the EPO disclaims all liability in this regard.

Patent documents cited in the description

• US 4960759 A, De Luca [0007] • US 5567689 A [0009] [0026] [0027] • US 5470838 A, von Borstel [0008] [0024] • US 5723449 A [0009] [0027] • US 5141943 A [0009] [0025]

Non-patent literature cited in the description

• CORNFORD et al. independent blood-brain barrier • MONTICONE GF et al. On the therapeutic use of the transport systems for nucleic acid precursors.Bio- nucleosides, cytidine and uridine, in some neurolog- chim. Biophys. Acta, 1975, vol. 349, 211-219 [0002] ical diseases. Minerva Med., 19 December 1966, vol. • DAWSON. Enzymic conversion of uridine nucleotide 57 (101), 4348-4352 [0006] to cytidine precursor by rat brain.J. Neurochem., • LOPEZ-COVIELLA et al. Evidence that 5’-cyti- 1968, vol. 15, 31-34 [0004] dinephosphocholine can affect brain phospholipid • BECROFT DM et al. Hereditary orotic aciduria: long- composition by increasing choline and cytidine plas- term therapy with uridine and a trial of uracil. J Pedi- ma levels. J. Neurochemistry, 1995, vol. 65, 889-894 atr., November 1969, vol. 75 (5), 885-891 [0006] [0020] • ROBERTS CA et al. Uridine anticonvulsant effects: • GINSBURG et al. Rodent models of cerebral selective control of nucleoside incorporation in exper- ischemia. Stroke, 1989, vol. 20, 1627-1642 [0021] imental epilepsy. Epilepsia, December 1974, vol. 15 (4), 479-500 [0006]

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