(19) &   

(11) EP 2 322 187 A2

(12) EUROPEAN PATENT APPLICATION

(43) Date of publication: (51) Int Cl.: 18.05.2011 Bulletin 2011/20 A61K 31/7072 (2006.01) A61K 31/14 (2006.01) A61K 31/685 (2006.01) A61P 25/28 (2006.01) (21) Application number: 10075660.0

(22) Date of filing: 30.07.1999

(84) Designated Contracting States: (72) Inventors: AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU • Watkins, Carol MC NL PT SE Cambridge, MA 02142 (US) • Wurtman, Richard J. (30) Priority: 31.07.1998 US 95002 P Boston, MA 02116 (US)

(62) Document number(s) of the earlier application(s) in (74) Representative: Maury, Richard Philip accordance with Art. 76 EPC: Marks & Clerk LLP 09173495.4 / 2 145 627 90 Long Acre 07116909.8 / 1 870 103 London 99937631.2 / 1 140 104 WC2E 9RA (GB)

(71) Applicant: Massachusetts Institute of Technology Remarks: Cambridge, Massachusetts 02139 (US) This application was filed on 01-10-2010 as a divisional application to the application mentioned under INID code 62.

(54) Use of in combination with choline for the treatment of memory disorders

(57) A preparation for use in the prevention and/or ageing, said preparation comprising (a) uridine or a uri- treatment of memory disorders, memory decline, cogni- dine source and (b) chholine, a choline precursor, a tive dysfunction, and/or slowing down or reversing brain- choline salt or ester, or a mixture thereof. EP 2 322 187 A2

Printed by Jouve, 75001 PARIS (FR) 1 EP 2 322 187 A2 2

Description art can dismiss them either as insignificant or irreproduc- ible. Moreover, Dawson himself states that he was una- FIELD OF THE INVENTION ble to recover a with spectrophotometric char- acteristics of and admits that his conclusions [0001] The present invention relates to methods of in- 5 were based on probabilistic guessing. Thus, the alleged creasing cytidine levels by administering an exogenous phenomenon observed by Dawson may have been due uridine source and in particular to the pharmacological to misinterpretation of an experimental artifact as it is use of said uridine or uridine source alone or in combi- now known that experimentally measurable cytidine can nation with other pharmaceutical substances in treating be easily confused with tyrosine, which is chemically un- certain neurological disorders. 10 related amino acid compound (see Fig. 1). [0005] Thus, even though an enzyme 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- 15 any doubt. Thus, these levels may be not sufficient to istration of uridine or uridine source to certain animals 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 20 ical modality. In addition, as it is the case with many other as administration of cytidine or cytidine precursors. How- enzymes 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 25 does not necessarily means that a similar process is much less efficient than uridine in crossing the blood- 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- 30 Consequently, the prior art is silent in regard to the use tabolism of compounds, enzymes are known 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 35 cogens such as UDP glucose and UTP glucose. Medical species lack this enzyme. However, according to EC (en- 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: long-term therapy with uridine and a trial [0004] The prior art relating to the process of uridine 40 of . J Pediatr. 1969 Nov; 75(5): 885-891 ). Other to cytidine conversion is also limited. Only one publica- 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 conversion of uridine nucleotide to cytidine nucleotide 45 1974 Dec; 15(4): 479-500 ). Most commonly, uridine is ( Dawson. Enzymic conversion of uridine nucleotide to 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 , 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 par- zyme does not appear to be sufficiently potent. As com- 50 ticular dual combination range from liver and kidney dis- pared to the initial, administered dose of uridine (consid- eases to a number of neurological and cerebrovascular ered as 100%), the highest levels of newly converted diseases but such uses are irrelevant to the present in- cytidine in vivo were 12.4% in the liver and 9% in the vention directed at the use of uridine without concomitant brain. The conversion rates in vitro were 5.4% in the liver use with cytidine. and 8.05% in the brain. Thus, maximum observed levels 55 [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 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

2 3 EP 2 322 187 A2 4 al., teach that the benefit of uridine is due to increase in polio syndrome, and muscular dystrophies are also pos- 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 5 as treated by combination therapy in which uridine is one the preferred embodiment of the instant invention is treat- 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 von10 the light of the prior art. Borstel et al., on November 28, 1995 discloses the meth- 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 15 covery that uridine administration in humans leads to in- use both forms of since it was not obvious crease in systemic and braincytidine. While certain meth- to them that uridine alone is effective. The absolute re- ods of treatment of certain neurological diseases using quirement of both cytidine and uridine was due to the uridine are known the prior art has heretofore failed to lack of knowledge and anticipation in the prior art that provide methods of treatment of diseases which are ob- uridine might convert into cytidine, especially in humans. 20 jects of this invention. These and other objects of the One skilled in the art will recognize that the disclosed invention will become more readily apparent from the fol- composition matter is different and diseases to be treated lowing description. are not the same as in the present invention. [0013] It is a general object of this invention to provide [0009] The U.S. Pat. Nos. 5,141,943 ; 5,567,689 ; and a novel method of treatment using uridine, uridine prod- 5,723,449 disclose various methods and compositions 25 rugs, and pharmaceutically derivatives thereof alone or to raise levels of uridine in the blood as useful for reducing in combination with compounds that are useful in increas- toxicity of pyrimidine nucleoside drugs such as AZT and ing uridine availability. The terms "uridine precursor" or 5-Fluouracil for AIDS and cancer therapy respectively. It "uridine source" or "uridine prodrug" are used inter- is apparent to anyone skilled in the art that these teach- changeably and as defined hereinafter mean com- ings have nothing in common with the present invention. 30 pounds, e.g., uridine salts or food products containing [0010] Although all of these patents and prior art ref- uridine, that transform into uridine upon administration erences disclose at least one or another aspect of the to a host such as human. instant invention none of them taught specifically that cy- [0014] The present invention also provides a method tidine levels can be raised in humans by administering of treating various neurological disorders comprising uridine or uridine source as useful for the treatment of 35 memory disorders associated with aging as well as mem- certain neurological or brain disorders. These disorders ory decline and cognitive dysfunction associated with comprise disorders associated with aging such as mem- pathological conditions like Alzheimer’s disease, Pick’s ory decline and age related decline in cognition functions. disease, Lewy Body disease, and/or dementias like Hunt- These disorders also comprise memory decline and re- ington’s disease and AIDS dementia. lated cognition dysfunction associated with pathological 40 [0015] The present invention also provides a method conditions like Alzheimer’s disease, Pick’s disease, Lewy of slowing down or reversing brain- aging process, which Body disease, and/or dementias like Huntington’s dis- may cause memory decline and cognitive dysfunction. ease and AIDS dementia. Other cognitive dysfunctions, [0016] Other uses of uridine therapy are provided such i.e., disorders of attention, alertness, concentration, fo- as treatment of cognitive dysfunction, i.e., disorders of cus, and dyslexia can also be treated. Other uses of uri- 45 attention, alertness, concentration, focus, and dyslexia. dine therapy can be imagined such as treatment of mood [0017] Other uses of uridine therapy are also provided and emotional disorders, e.g., mania, depression, stress, such as treatment of mood and emotional disorders, e.g., panic, anxiety, insomnia, dysthemia, psychosis, season- mania, depression, stress, panic, anxiety, insomnia, dys- al effective disorders and bipolar disorders. Neurological themia, psychosis, seasonal effective disorders and bi- diseases like ataxias, including Friedreich’s ataxia and 50 polar disorders. movement disorders like tardive dyskinesia can also be [0018] It is also an object of this invention to provide treated. Method of treating stroke, cerebral thrombosis, methods for treating other neurological diseases like ischemia, and related cerebrovascular diseases result- ataxias, including Friedreich’s ataxia. ing from hypoxia as well as behavioral and neurological [0019] It is also an object of this invention to provide syndromes seen after brain trauma, spinal cord injury 55 methods for treating strokes, cerebral thrombosis, and/or anoxia can be also imagined. Methods of treating ischemia, and related cerebrovascular diseases result- diseases of the peripheral nervous system, e.g., neu- ing from hypoxia. romuscular disorders like myasthenia gravis, the post- [0020] Methods of treating behavioral and neurological

3 5 EP 2 322 187 A2 6 syndromes seen after brain trauma, spinal cord injury and other known in the art compounds or mixtures thereof and/or anoxia are also provided. involved in phospholipid synthesis. [0021] The present invention also provides a method of treating patients with movement disorders like tardive BRIEF DESCRIPTION OF THE DRAWINGS dyskinesia. 5 [0022] Methods of treating diseases of the peripheral [0028] nervous system, e.g., neuromuscular disorders like myasthenia gravis, the post-polio syndrome, and mus- Fig. 1 illustrates the coincidence of cytidine and ty- cular dystrophies are also provided. rosine peaks (6.59) when tested by a standard HPLC [0023] It is also an object of this invention to provide 10 method. methods for treating other diseases known in the art and Fig. 2 illustrates distinct cytidine (3.25) and tyrosine involving or dependent on cholinergic or uridine/ cytidine (2.92) peaks when tested by a modified HPLC meth- metabolic pathways. od, which utilizes elution buffer with low methanol. [0024] Methods of treating neurological diseases as- Fig. 3 shows the ratio of uridine (100%) to cytidine sociated with dopaminergic pathway, e.g., schizophrenia 15 in plasma after oral administration of 250 milligram and Parkinson’s disease as treated by combination ther- per kg of body weight (mg/kg) of uridine. apy in which uridine is one of constituents are provided. Fig. 4 shows the ratio of uridine (100%) to cytidine [0025] In another aspect, the invention provides addi- in the brain after oral administration of 250 milligram tional pharmaceutical compositions that can inhibit met- per kg of body weight (mg/kg) of uridine. abolic degradation or clearance of uridine as compounds 20 enhancing uridine bioavailability. Among such com- DESCRIPTION OF THE PREFFERED EMBODIMENTS pounds are uridine phosphorylase inhibitors like 5- benzyl or derivatives thereof. Among such com- [0029] The present invention concerns methods and pounds are also compounds competing with uridine in compositions for increasing systemic and brain levels of renal transport mechanism. They comprise compounds 25 cytidine in a human patient via administration of uridine like L-uridine, L-2’,3’-dideoxyuridine, and D- 2’,3’-dideox- or uridine source alone. Also methods and compositions yuridine. Among such compounds are also uridine se- are contemplated which encompass the use of uridine cretion inhibiting compounds like or hexobend- or uridine source in combination with drugs that increase ine. uridine availability. Among such drugs are drugs that act [0026] Choline is involved in metabolism and lipid30 as uridine phosphorylase inhibitors like benzyl barbitu- transport and is a component of a number of important rate or derivatives thereof. Among such drugs are also biological compounds including the membrane phos- drugs that act as uridine secretion inhibiting compounds pholipids like lecithin and sphingomyelin. Choline is also likedilazep or hexobendine. Among such drugs are drugs a precursor of acetylcholine - one of most important neu- that act as uridine renal transport competitors like L-uri- rotransmitters. Although a required nutrient for several 35 dine, L-2’,3’-dideoxyuridine, and D-2’,3’-dideoxyuridine. animal species, choline is not currently designated as Also methods and compositions are disclosed that are essential for humans. However, recent clinical studies beneficial to a human patient in need thereof and which show it to be essential for normal liver function. Addition- act in synergy with uridine in generation of phospholipids ally, a large body of evidence from the fields of molecular involved in brain cell membrane formation and repair. and cell biology shows that certain phospholipids play a 40 More specifically choline- based compounds are contem- critical role in generating second messengers for cell plated as compounds acting in synergy with uridine or membrane signal transduction. This process involves a uridine source. Among them are choline, choline salts or cascade of reactions that translate an external cell stim- esters, such as choline bitartrate or stearate or the like, ulus such as a hormone or growth factor into a change or compound that dissociate to choline, such as sphin- in cell transport, metabolism, growth, function, or gene 45 gomyelin, cytidine-diphospho-choline or citicoline or expression. Disruptions in phospholipid metabolism can CDP-choline,acylglycerophosphocholines, e.g., lecithin, interfere with this process and may underlie certain dis- lysolecithin, glycerophosphatidylcholine, mixtures there- ease states such as cancer and Alzheimer’s disease. of or the like. However, choline alone is not useful as a therapeutical [0030] Neurological diseases associated with modality. In light of the instant invention choline or choline 50 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. [0027] It is thus a further object of this invention to es- [0031] Other neurological conditions that were sug- tablish a synergy between uridine and various com- gested to be treatable by uridine like epilepsy, seizures, pounds affecting cholinergic pathway and/or phospholi- 55 and convulsions are excluded. pid metabolism. Among them are CDP-choline, choline, [0032] Specific medical uses of uridine in combination choline salts, lecithin or phosphatidylcholine, phosphati- with cytidine known in the prior art are also excluded. dylethanolamine, various fatty acids, e.g., linoleic acid, [0033] The following examples illustrate the invention

4 7 EP 2 322 187 A2 8 and are not intended to be limiting unless otherwise spec- the brain is totally unexpected and constitutes the basis ified. for the present invention.

EXAMPLE 1. EXAMPLE 3 5 [0034] In this example a new method is established [0036] In Example 3 a clinical study is carried out with that overcomes the problemof the coincidence of cytidine the goal of treating memory disorders and cognitive dys- and tyrosine peaks when tested by a standard HPLC functions associated with aging as well as memory de- method for measuring various nucleosides in biological cline and cognitive dysfunction associated with patho- fluids (see Fig. 1). By using the standard HPLC method, 10 logical conditions like Alzheimer’s disease, Pick’s dis- one can, however, easily distinguish the uridine peak ease, Lewy Body disease, and/or dementias like Hunt- from the cytidine peak. Detailed description of the HPLC ington’s disease and AIDS dementia. Patients with non- method can be found for example in Lopez-Coviella et pathological dementia associated with aging are also in- al., (Evidence that 5’-cytidinephosphocholine can affect cluded. Oral doses of uridine alone ranging from 5 mg to brain phospholipid composition by increasing choline15 50,000 mg are administered daily to five male and five and cytidine plasma levels. J. Neurochemistry 65: female patients suffering from one of the diseases listed 889-894, 1995 ). Modified HPLC is carried out in a same above. The adjustment in dosage to select optimally ef- way as standard HPLC except that elution buffer contains fective pharmaceutical dose is a routine procedure well low amount of methanol (0.1%) instead of formic acid known to the practitioner skilled in the relevant art. The and as a result cytidine can be distinguished from unre- 20 terms"therapeutically" or "pharmaceutically"or "pharma- lated compound tyrosine (Fig. 2). This method is useful cologically effective dose" of a drug as used hereinafter in distinguishing cytidine from masking effect of amino means the amount (dosage) of the drug that provides acid tyrosine, which may concomitantly be present in desired clinical effect in at least 10% of treated tested biological fluid, e.g., plasma or cerebrospinal fluid patients" population. (CSF). Due to the overlap between cytidine and tyrosine 25 [0037] Various other uridine-based compounds other it is very likely that the results of all the prior art studies than uridine itself serve as uridine source or uridine pre- dealing with cytidine measurement, including present in- cursors. These are uridine-rich food or dietary products ventors’ own studies supra, were interpreted incorrectly. like algae; salts of uridine like uridine phosphates, acylat- ed uridine or the like. These also include compounds, EXAMPLE 2. 30 like CDP-choline, though structurally unrelated to uridine are able, nevertheless, to raise uridine levels in treated [0035] Gerbils rather than rats or other rodents are se- subjects. If required by the exigency of the therapy, ther- lected for this example, as the pyrimidine metabolism of apeutically or pharmacologically effective doses of CDP- said gerbils is closer to humans. For practical and ethical choline are also administered since it is now known that reasons humans cannot always be used for certain ex- 35 administration of said drug raises uridine but not cytidine perimental studies and those skilled in the art generally levels and as such CDP- choline or citicoline is by defini- recognize that the gerbil model is equivalent to a human tion the uridine source. model. Indeed, gerbils are the choice model for certain [0038] If required by the exigency of the therapy, ther- human diseases and brain disorders such as cerebral apeutically or pharmacologically effective doses of acyl ischemia (Ginsburg et al., Rodent models of cerebral 40 derivatives of uridine or mixtures thereof like those dis- ischemia. Stroke 20: 1627-1642, 1989 ). Grerbils are giv- closed in U.S. Pat. No. 5,470,838 are also administered. en orally uridine and 60 minutes later plasma and brain [0039] If required by the exigency of the therapy, ther- levels of cytidine and uridine are measured by modified apeutically or pharmacologically effective doses of urid- HPLC method described in Example 1. The Fig. 3 shows ine phosphorylase inhibitors like 5-benzyl barbiturate de- the relative ratio between uridine and cytidine levels in 45 rivatives or mixtures thereof as disclosed in U.S. Pat. No. plasma after oral administration of 250 milligram per kg 5,141,943 are also administered. of body weight (mg/kg) of uridine. The Fig. 4 shows the [0040] If required by the exigency of the therapy, ther- relative ratio between uridine and cytidine levels in the apeutically or pharmacologically effective doses of urid- brain after oral administration of 250 mg/kg of uridine. ine secretion inhibiting compounds like dilazep, hex- These results indicate that the metabolic processing of 50 obendine, or mixtures thereof as disclosed in U.S. PAT. uridine in the brain is different than systemic processing No. 5,567,689 are also administered. of uridine in plasma. The results also indicate that uridine, [0041] If required by the exigency of the therapy, ther- when transported into the brain, is readily converted to apeutically or pharmacologically effective doses of com- cytidine and this conversion is more efficient in the brain pounds which compete with uridine in kidney clearance than in plasma. Similar experiments are also carried out 55 like L-uridine, L-2’,3’- dideoxyuridine, and D- 2’,3’-dideox- in humans wherein instead of measuring brain levels of yuridine or mixtures thereof as disclosed in U.S. Pat. Nos. nucleosides the CSF levels are measured. The finding 5,723,449 and 5,567,689 are also administered. Thera- that uridine is readily converted to cytidine especially in peutically or pharmacologically effective doses of uridine

5 9 EP 2 322 187 A2 10 as defined herein are also doses that produce blood or [0045] In Example 5 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 ([micro]M) and Imillimole (mM). In general, therapeuti- study of Example 3 except that patients enrolled in this cally or pharmacologically effective doses as defined study are patients with mood and emotional disorders, herein are also doses of drugs combination, which pro- 5 e.g., mania, depression, stress, panic, anxiety, insomnia, duce desired effect in at least 10% of treated patients= dysthemia, psychosis, seasonal effective disorders and population. Doses are administered either as a single bipolar disorders. dose or divided in several doses. Drugs are administered [0046] In Example 6 a clinical study is carried out, orally such as in tablet, capsule or liquid form or parenter- which by its design and principles is similar to clinical ally by intravenous, intramuscular or subcutaneous in- 10 study of Example 3 except that patients enrolled in this jection. study are patients with neurological diseases like ataxias, [0042] When necessary and as required by the exi- including Friedreich’s ataxia. gency of the therapy uridine is administered in combina- [0047] In Example 7 a clinical study is carried out, tion with other compounds that act either synergistically which by its design and principles is similar to clinical or in additive manner. This reduces the therapeutic dose 15 study of Example 3 except that patients enrolled in this of administered drugs, thereby reducing potential unde- study are patients with movement disorders like tardive sirable side effects and frequency of drug administration. dyskinesia. Compounds that act in such a manner are chemical sub- [0048] In Example 8 a clinical study is carried out, stances participating in cholinergic metabolism. For ex- which by its design and principles is similar to clinical ample, compounds administered along with uridine are 20 study of Example 3 except that patients enrolled in this following choline-based compounds: choline, choline study are patients with strokes, cerebral thrombosis, salts or esters, such as choline bitartrate or stearate or ischemia, and related cerebrovascular diseases result- the like, or compound that dissociate to choline, such as ing from hypoxia. sphingomyelin, cytidine-diphospho-choline or citicoline [0049] In Example 9 a clinical study is carried out, or CDP-choline, acylglycerophosphocholines, e.g., leci- 25 which by its design and principles is similar to clinical thin, lysolecithin, glycerophosphatidylcholine, mixtures study of Example 3 except that patients enrolled in this thereof or the like. The choline or compound that disso- study are patients with behavioral and neurological syn- ciates into choline is administered so that a choline level dromes seen after brain trauma, spinal cord injury and/or of at least about 20-30 nanomoles and usually between anoxia. 10 and 50 nanomoles is attained in patient=s blood or 30 [0050] In Example 10 a clinical study is carried out, brain. which by its design and principles is similar to clinical [0043] The pharmacologically effectivedoses are with- study of Example 3 except that patients enrolled in this in about 20 mg and 50 g/day range, preferably between study are patients with diseases of the peripheral nervous about 100 mg and 10 g/day. Doses are administered ei- system, e.g., neuromuscular disorders like myasthenia ther as a single dose or divided in several doses, e.g., 35 gravis, the post-polio syndrome, and muscular dystro- 10 mg to 1 g/ cap or tab. The minimal duration of the phies. therapy is at least one day but longer periods of time are [0051] In Example 11 a clinical study is carried out, usually required according to the exigency of the therapy. which by its design and principles is similar to clinical If needed, the usual time period spans from one day to study of Example 3 except that patients enrolled in this the period of lifetime. When these compounds are not 40 study are patients with neurological diseases associated available in pure form the active ingredient comprises at with dopaminergic pathway, e.g., schizophrenia and Par- leastabout 20-30 percentof the weight of thepreparation. kinson’s disease and said diseases are treated by com- The clinical study is continued for at least 1 day or longer bination therapy in which uridine is one of constituents. as required by the exigencies of the therapy. In general, [0052] In Example 12 a clinical study is carried out, the dose administered, the frequency of administration 45 which by its design and principles is similar to clinical and the duration of the treatment will vary as a function study of Example 3 except that patients enrolled in this ofthe condition of the patient and is determined according study are patients with other diseases known in the art to standard clinical procedures known to the practitioner and involving or dependent on cholinergic or uridine/cy- skilled in the relevant art. tidine metabolic pathways. 50 [0053] When possible clinical studies as disclosed in EXAMPLES 4-12 any preceding examples are preceded by in vivo studies in animal models, e.g., gerbil model, according to proce- [0044] In Example 4 a clinical study is carried out, dures established in the art. which by its design and principles is similar to clinical [0054] While methods of treatment and pharmaceuti- study of Example 3 except that patients enrolled in this 55 cal compositions recited in one of above examples have study are patients with cognitive dysfunction, i.e., disor- been described in detail, it will be apparent to those skilled ders of attention, alertness, concentration, focus, and in the art that these embodiments are exemplary rather dyslexia. than limiting and the true scope of the invention is that

6 11 EP 2 322 187 A2 12 defined within the attached claims. effective dose of at least uridine or a uridine source and other compounds. 1. A method of treating a human with neurological 17. The method according to item 16 wherein said disorders comprising administering therapeutically neurological disorders are memory disorders. effective doses of uridine or a uridine source. 5 18. The method according to item 17 wherein said 2. The method according to item 1 wherein said ther- memory disorders comprise memory decline asso- apeutically effective dose of uridine or a uridine ciated with brain aging. source is administered for at least one day. 19. The method according to item 17 wherein said 3. The method according to item 1 wherein said neu- memory disorders are selected from the group com- rological disorders are memory disorders. 10 prising Pick’s disease, Lewy Body disease, and/or 4. The method according to item 3 wherein said dementias like Huntington’s disease and AIDS de- memory disorders comprise memory decline asso- mentia. ciated with brain aging. 20. The method according to item 16 wherein said 5. The method according to item 3 wherein said neurological disorders are cognitive dysfunctions memory disorders are selected from the group com- 15 comprising lack of attention, alertness, concentra- prising of Pick’s disease, Lewy Body disease, and/or tion, focus, or dyslexia. dementias like Huntington’s disease and AIDS de- 21. The method according to item 16 wherein said mentia. neurological disorders are emotional disorders com- 6. The method according to item 1 wherein said neu- prising mania, depression, stress, panic, anxiety, rological disorders are cognitive dysfunctions com- 20 dysthemia, psychosis, seasonal effective disorders prising lack of attention, alertness, concentration, fo- and bipolar disorders. cus, or dyslexia. 22. The method according to item 16 wherein said 7. The method according to item 1 wherein said neu- neurological disorders are selected from the group rological disorders are emotional disorders compris- consisting of ataxia or Friedreich’s ataxia. ing mania, depression, stress, panic, anxiety, dys- 25 23. The method according to item 16 wherein said themia, psychosis, seasonal effective disorders and neurological disorders are movement disorders bipolar disorders. comprising tardive dyskinesia. 8. The method according to item 1 wherein said neu- 24. The method according to item 16 wherein said rological disorders are selected from the group con- neurological disorders are cerebral thrombosis, sisting of ataxia or Friedreich’s ataxia. 30 ischemia, and related cerebrovascular diseases re- 9. The method according to item 1 wherein said neu- sulting from hypoxia. rological disorders are movement disorders com- 25. The method according to item 16 wherein said prising tardive dyskinesia. neurological disorders are behavioral and neurolog- 10. The method according to item 1 wherein said ical syndromes. neurological disorders are cerebral thrombosis,35 26. The method according to item 25 wherein said ischemia, and related cerebrovascular diseases re- behavioral and neurological syndromes comprise sulting from hypoxia. those seen after brain trauma, spinal cord injury 11. The method according to item 1 wherein said and/or anoxia. neurological disorders are behavioral and neurolog- 27. The method according to item 16 wherein said ical syndromes. 40 neurological disorders are peripheral nervous sys- 12. The method according to item 11 wherein said tem disorders comprising neuromuscular disorders behavioral and neurological syndromes comprise like myasthenia gravis, the post-polio syndrome, and those seen after brain trauma, spinal cord injury muscular dystrophies. and/or anoxia. 28. The method according to item 16 wherein said 13. The method according to item 1 wherein said 45 neurological disorders selected from the group con- neurological disorders are peripheral nervous sys- sisting of schizophrenia and Parkinson’s disease. tem disorders comprising neuromuscular disorders 29. The method according to item 16 wherein said like myasthenia gravis, the post-polio syndrome, and other compound is choline. muscular dystrophies. 30. The method according to item 16 wherein said 14. The method according to item. 1 wherein said 50 other compounds are choline salt or stearate select- uridine is combined with pharmaceutically accepta- ed from the group consisting of choline chloride, ble source of uridine. choline bitartrate, choline stearate or mixtures there- 15. The method according to item I wherein said uri- of. dine or pharmaceutically acceptable source of urid- 31. The method according to item 16 wherein said ine is administered in dosages of between 100 mg 55 other compounds are choline precursors selected and 20 grams per day. from the group consisting of sphingomyelin, cytidine- 16. A method of treating a human with neurological diphospho-choline or citicoline or CDP-choline, an disorders comprising administering therapeutically acylglycerophosphocholine, e.g., lecithin, lysoleci-

7 13 EP 2 322 187 A2 14

thin, glycerophosphatidylcholine, fatty acids, or mix- 7. The preparation according to any one of the preced- tures thereof. ing claims, wherein the choline precursor is sphin- 32. The method as in one of items 29-31 wherein a gomyelin, cytidine-diphospho-choline, acylglycero- choline level of at least about 20-30 nanomoles and phosphocholine (AGP-choline), phosphatidylcho- usually between 10 and 50 nanomoles is attained in 5 line (PC), or a combination thereof. patient=s blood or brain. 33. The method according to item 16 wherein said 8. The preparation according to any one of the preced- other compounds are uridine phosphorylase inhibi- ing claims, whrein the choline salt is choline chloride, tors comprising benzyl barbiturate or derivatives choline bitartrate, choline stearate or mixtures there- thereof. 10 of. 34. The method according to item 16 wherein said other compounds are uridine secretion inhibiting 9. The preparation according to any one of the preced- compounds comprising dilazep or hexobendine. ing claims, wherein said uridine or uridine source is 35. The method according to item 16 wherein said administered in dosages between about 10 mg and other compounds are uridine renal transport com- 15 10 grams per day. petitors selected from the group consisting of L-uri- dine, L-2’,3’-dideoxyuridine, and D-2’,3’-dideoxyuri- 10. The preparation according to any one of the preced- dine. ing claims, wherein said uridine comprises uridine 36. The method according to item 16 wherein said salts or a food product containing uridine. other compound is a mixture of compounds from any 20 one of items 29-31 or 33-35. 11. Use of a preparation in the manufacture of a com- 37. The method according to item 16 wherein said position for the prevention and/or treatment of mem- uridine or a uridine source are administered in dos- ory disorders, memory decline, cognitive dysfunc- ages of between 10 mg and 10 grams per day. tion, and/or slowing down or reversing brain- ageing, 38. The method according to item 16 wherein said 25 said composition comprising (a) uridine or a uridine therapeutically effective dose of uridine or a uridine source and (b) choline, a choline precursor, a choline source in combination with other drugs is adminis- salt or ester, or a mixture thereof. tered for at least one day. 12. Use according to claim 11, wherein said cognitive 30 dysfunction is associated with aging or with patho- Claims logical conditions.

1. A preparation for use in the prevention and/or treat- ment of memory disorders, memory decline, cogni- tive dysfunction, and/or slowing down or reversing 35 brain-ageing, said preparation comprising (a) uridine or a uridine source and (b) choline, a choline precur- sor, a choline salt or ester, or a mixture thereof.

2. The preparation according to claim 1, wherein said 40 cognitive dysfunction is associated with aging or with pathological conditions.

3. The preparation according to claim 1, wherein said memory disorders are associated with aging and 45 age-related decline in cognition functions.

4. The preparation according to claim 1, wherein said memory disorder is Alzheimer’s disaese, Pick’s dis- ease, Lewy Body disease and/or dementias. 50

5. The preparation according to claim 4, wherein said dementias comprise Huntington’s disease or AIDS dementia. 55 6. A preparation comprising (a) uridine or a uridine source and (b) choline, a choline precursor, a choline salt or ester, or a mixture thereof.

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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] [0040] [0041] • US 5470838 A, von Borstel [0008] [0038] • US 5723449 A [0009] [0041] • US 5141943 A [0009] [0039]

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’-cytidinephos- 1968, vol. 15, 31-34 [0004] phocholine can affect brain phospholipid composition • Becroft DM et al. Hereditary orotic aciduria: by increasing choline and cytidine plasma levels. J. long-term therapy with uridine and a trial of uracil. J Neurochemistry, 1995, vol. 65, 889-894 [0034] Pediatr., November 1969, vol. 75 (5), 885-891 [0006] • Ginsburg et al. Rodent models of cerebral ischemia. • Roberts CA et al. Uridine anticonvulsant effects: se- Stroke, 1989, vol. 20, 1627-1642 [0035] lective control of nucleoside incorporation in experi- mental epilepsy. Epilepsia, December 1974, vol. 15 (4), 479-500 [0006]

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