(19) TZZ ¥ __T (11) EP 2 322 187 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) 14.05.2014 Bulletin 2014/20 A61K 31/685 (2006.01) A61P 25/28 (2006.01) (21) Application number: 10075660.0 (22) Date of filing: 30.07.1999 (54) Use of uridine in combination with choline for the treatment of memory disorders Verwendung von Uridin in Kombination mit Cholin zur Behandlung von Gedächtnisstörungen Utilisation de l’uridine en combinaison avec la choline pour le traitement des maladies de la mémoire (84) Designated Contracting States: (56) References cited: AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU WO-A-97/45127 CH-A5- 680 334 MC NL PT SE DE-A1- 2 508 474 DE-A1- 2 629 845 DE-U1- 9 412 374 US-A- 4 221 784 (30) Priority: 31.07.1998 US 95002 P US-A- 4 994 442 US-A- 5 567 689 US-A- 5 700 590 (43) Date of publication of application: 18.05.2011 Bulletin 2011/20 • CACABELOSR ET AL: "THERAPEUTIC EFFECTS OF CDP-CHOLINE IN ALZHEIMER’S DISEASE (62) Document number(s) of the earlier application(s) in COGNITION, BRAIN MAPPING, accordance with Art. 76 EPC: CEREBROVASCULAR HEMODYNAMICS, AND 09173495.4 / 2 145 627 IMMUNE FACTORS", ANNALS OF THE NEW 07116909.8 / 1 870 103 YORK ACADEMY OF SCIENCES, NEW YORK 99937631.2 / 1 140 104 ACADEMY OF SCIENCES, NEW YORK, NY, US, 1 January 1996 (1996-01-01), pages 399-403, (73) Proprietor: Massachusetts Institute of Technology XP008065562, ISSN: 0077-8923 Cambridge, Massachusetts 02142-1601 (US) • SPIERS P A ET AL: "CITICOLINE IMPROVES VERBAL MEMORY IN AGING", ARCHIVES OF (72) Inventors: NEUROLOGY, AMERICAN MEDICAL • Watkins, Carol ASSOCIATION, CHICAGO, IL, US, vol. 53, no. 5, Cambridge, MA 02142 (US) 1 May 1996 (1996-05-01), pages 441-448, • Wurtman, Richard J. XP008028412, ISSN: 0003-9942 Boston, MA 02116 (US) • WEISS G B: "Metabolism and actions of CDP- choline as an endogenous compound and (74) Representative: Korn, Richard Mervyn administered exogenously as citicoline.", LIFE Pearl Cohen Zedek Latzer Baratz UK LLP SCIENCES 1995 LNKD- PUBMED:7869846, vol. 15 Old Bailey 56, no. 9, 1995, pages 637-660, XP002640081, London EC4M 7EF (GB) ISSN: 0024-3205 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 2 322 187 B1 Printed by Jouve, 75001 PARIS (FR) 1 EP 2 322 187 B1 2 Description ble to recover a nucleotide with spectrophotometric char- acteristics of cytidine and admits that his conclusions [0001] The present invention relates to the use of uri- were based on probabilistic guessing. Thus, the alleged dine or a uridine source in combination with choline, a phenomenon observed by Dawson may have been due choline precursor, a choline salt or ester, or a mixture 5 to misinterpretation of an experimental artifact as it is thereof in the manufacture of a composition for treating now known that experimentally measurable cytidine can cognitive dysfunction associated with aging, and/or for be easily confused with tyrosine, which is chemically un- slowing down or reversing brain-aging. related amino acid compound (see Fig. 1). [0005] Thus, even though an enzyme catalyzing the DESCRIPTION OF THE RELATED ART 10 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 warrant practical exploitation for clinical application. In- comprising human patients, leads to increased levels of 15 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- enzymes and metabolic pathways, this particular en- ever, the potential benefit of uridine or uridine source 20 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- present in another animal, e.g., man. Based on that one brain barrier (Cornford et al., Independent blood-brain 25 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 of the process of uridine to cytidine conversion for any in the art, such as cytidine deaminase (EC 3.5.4.5), which 30 meaningful application. converts cytidine into uridine. Cytidine deaminase can [0006] Uridine is a pyrimidine nucleoside 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- uses of uridine alone are limited to treatment of genetic zyme classification) list there are no known examples of 35 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 uracil. 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 40 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 used in combination with cytidine (Monticone GF, et al., cytidine precursor by rat brain. J. Neurochem. 15:31-34, 45 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- diseases to a number of neurological and cerebrovascu- ered as 100%), the highest levels of newly converted 50 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 al., on October 2, 1990 discloses the pharmacological view all these figures are within the range of a typical 55 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 ible. Moreover, Dawson himself states that he was una- cholecystokinin levels in the brain, which in turn improves 2 3 EP 2 322 187 B1 4 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- of constituents. ment of neurological disorders unrelated to schizophre- 5 [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.