AU671995B2 - Purine derivatives - Google Patents

Description

OPI DATE 13/12/93 AOJP DATE 24/02/94 APPLN. ID 40612/93 II1111 111111 PCT NUMBER PCT/DK93/00158 II I 11111 1 1111111111 INI AU934061 2 1. IRNN'I IONAL APIPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (51) International Patent Classification 5 (11) International Publication Number: WNO 93/23418 C07H 19/16, 19/167, A61IK 31/70 Al (43) International Publication Date., 25 November 1993 (25.11,93) (21) International Application Number: PCT/DK93/00l58 Published Withi international searc/h report.

(22) International Filing Date: 12 May 1993 (12.05.93) Priority data: 0626/92 14 May 1992 (14.05,92) DK6 74" zry (71) Applicant: NOVO NORDISK A/S [DK/DK]; Novo Allc, DK-2800 Bagsvzerd tkDK), (72) Inventors: KNUTSEN, Lars, Jacob, Stray ;Aldersrovcj 7, DK-2950 Vedb~k LAU, Jesper ;Rosenvenget 3, DK-2530 Farum (DK).

(81) Designated States: AU, BG, CA, CZ, Fl, HU, JP, KR, NO, NZ, PL, PT, RO, RU, SK, UA, European patent (AT, BE, CH, DE, DK, ES, FR, GB, GR, 113, IT, LU, MC, N L, PT, S E).

(54)Title: PURINE DERIVATIVES R 2

YY

RZR

HO OH (57) Abstract A compound or formula or a pharmaceutically acceptable salt thereof, wherein X ik hydrogen, amino, halogen. hN.

droxv. lower alkox or lower alkyl and R1 is wherein Y is rnethylene or a valence bond, R- andI R 5 are 1-1 or lower, straight or branched alkyl. R-1 is H or lower alkyl. or R2 and R-1 can together form a cyclobutyl, cyclopentyl, cyclohexyl or phenyl ring. Z is oxygen, methylene, sulphur, sulphonyl or a valence bond, R 4 ik 1-1, lower alkyl, aralkyl, a mono or bicyclic ar('aat.

ic system optionally su~bstituted with various groups. The compounds base been found useful for treating central nervous system ailments, WO 93/23418 PCT/DK93/00158 Purine derivatives The present invention relates to a method for treating ischaemia, epilepsy and pain, to compounds for use in such a method and to pharmaceutical compositions containing the said compounds.

Background of the Invention Adenosine can be considered to be a hormone which has been shown to have a number of significant effects on the mammalian central nervous system (CNS) [Annual Reports in Medicinal Chemistry, 1988, 23, 39-48; International Review of Neurobiology (Smythies, J.R. and Bradley, R.J., eds.) Academic Press Inc., 1985, 27, 63-139], especially under conditions of neuronal stress where the compound appears to act as an endogenous neuroprotectant (Progress in Neurobiology, 1988, 31, 85-108, Trends in Pharmacological Sciences, 1988, 9, 193-194). For example, the concentration of adenosine has been demonstrated to rise greatly in certain brain regions following epileptic seizures or conditions of neuronal ischaemia/anoxia (Brain Research 1990, 516, 248-256), It has been established for some years now that centrally acting adenosine receptor agonists or compounds which increase extracellular adenosine levels can exhibit what is termed neuromodulator activity. Such substances influence the release of neurotransmitters in regions of the central nervous system (Annual Review of Neuroscience, 1985, 8, 103-124; Trends in Neurosciences, 1984, 164-168), with particular inhibitory effects on the release of the excitatory amino acid glutamic acid (glutamate) (Nature, 1985, 316, 148-150, Journal of Neurochemistry, 1992, 58, 1683-1689).

There are several CNS ailments for which this adenosine receptor mediated WO 93/23418 PCT/DK93/00158 -2neuromodulator activity could be of clear therapeutic benefit. Examples of these would include the treatment of convulsive disorders (European Journal of Pharmacology, 1991, 195, 261-265; Journal of Pharmacology and Experimental Therapeutics, 1982, 220, 70-76), prevention of neurodegeneration under conditions of brain anoxia/ischaemia (Neuroscience Letters, 1987, 83, 287-293; Neuroscience, 1989, 30, 451-462; Pharmacology of Cerebral Ischaemia 1990, (Kriegelstein, J. and Oberpichler, Eds., Wissenschaftliche Verlagsgesellschaft mbH: Stuttgart, 1990, pp 439-448) or the use of a purinergic agent in the treatment of pain (European Journal of Pharmacology, 1989, 162, 365-369; Neuroscience Letters, 1991, 121, 267-270).

Adenosine receptors represent a subclass (P1) of the group of purine nucleotide and nucleoside receptors known as purinoreceptors. This subciass has been further classified into two distinct receptor types which have become known as Al and A2. Extensive research has been carried out in a quest to identify selective ligands at these sites [see, for example, Comprehensive Medicinal Chemistry, Volume 3, (Hansch, Sammes, P.G.

and Taylor, Eds,, Pergamon Press PLC: 1990, pp 601-642)]. Selective ligands exist for Al and A2 adenosine receptors and the structure-activity relationships of the various reference ligands have been reviewed (Biochemical Pharmacology, 1986, 35, 2467-2481) together with their therapeutic potential (Journal of Medicinal Chemistry, 1992, 35, 407-422). Among the known adenosine receptor agonists most selective for the Al receptor over the A2 receptor are the examples where the adenine nucleus is substituted with a cycloalkyl group on the amino function, for example N-cyclopentyladenosine and N-cyclohaxyladenosine (Journal of Medicinal Chemistry, 1985, 28, 1383-1384) or 2-chldro-N-cyclopentyladenosine (Naunyn-Schmiedeberg's Arch. Pharmacol. 1988, 337, 687-689).

However, these ligands are found to possess undesirable effects as to WO 93/23418 PCT/DK93/60158 -3influence upon the cardiovascular system, rendering them unsuitable for the treatment of CNS disorders such as cerebral ischaemia, epilepsy and pain.

GB 1,143,150 (equivalent to USP 3,551,409) and GB 1,123,245 disclose a number of adenosine derivatives, having interesting cardiac and circulatory actions.

In EP 322,242A, a new use, as "agents to reduce plasma free fatty acid concentration or reducing heart rate and condition" is claimed for the compounds listed below as well as physiologically acceptable salts and solvates thereof: S, trans)-2-hydroxycyclopentyl] adenosine trans)-2-hydroxycyclopentyl]adenosine N-[(trans)-4-hydroxycyclohexyl]-2-methyladenosine N-[(cis)-4-hydroxycyclohexyl]adenosine N-[(cis)-2-hydroxycyclopentyl]adenosine N-[(trans)-3-hydroxycyclohexyl]adenosine N-[28-hydroxy-2-methylcyclopentyl]adenosine and N-[(cis)-2-hydroxycyclohexyl]adenosine Description of the Invention It has now been discovered that a selected group of adenosine derivatives, some of which are claimed in GB 1,143,150, has potential therapeutic utility for treating central nervous system ailments such as cerebral ischaemia, epilepsy and pain in humans. They have a clear CNS effect in relevant animal models at the same time as having a superior side-effect profile with respect to cardiovascular properties. In addition, the compounds have utility within myocardial ischaemia. Specifically, the following compounds possess therapeutic utility within the above-mentioned CNS indications: WO 93/23418 WO 9323418PC'/DK93/OO1 58 -4- 2-Chloro-N-(l -phenoxy-2-propyl)adenosinie 2-Chloro-N-((R) -1 -phenoxy-2-propyl) adenosine 2-Chloro-N- -1 -phenoxy-2-propyl] adenosine 2-Chloro-N-(2-phenoxyethyl) adenosine 2-Chloro-N-[(R) -1 -phenyl-2-propyl] adenosine 2-Chloro-N-(1 -phenyl-3-butyl) adenosine N-(1 -Phenoxy-2-propyl) adenosine 2-Amino-NA-(l -phenoxy-2-propyl)adenosine Nj-[ (iS, trans) -2-Hydroxycyclopentyl) adenosine NA-[(1 R, trans) -2-Hydroxycyclopentyl] adenosine 2-Chloro-N-(cis-2-phenoxycyclopentyl)adenosine trans-2-Chloro-N- (2-phenoxycyclopentyl) adenosine 2-Chl'oro-N-[ U' -1 -hydroxy-2-propyl] adenosine 2-Chloro-N-L 1-phenylthio-2-propyl]adenosine 2-Chloro-N- -(4-fluorophenoxy)-2-propyl] acenosine 2-Chloro-N-[(R)-2-phenoxy-1 -propyl) adenosine 2-Chloro- N- (phenylmethoxy) ethyl] adenosine 2-Fluoro-N-[(R)-1 -phenox, zwoopyfladenosine 2-Methoxy-N-[(R)-l -phen 2-propyijadenosine N-(2-Methoxyethyl)adenosine 2-Chloro-N-[(2-methoxyphenyl)methyl] adenosine 2-Chloro-N-[(R) -3-methyl-i -phenoxy-2-butyl~adenosine 2-Chloro-N-[(R)-l -(2-(2-propyloxy)phenoxy)-2-propyl] adenosine 2-Chloro-N- -1 -phenylsulphonyl-2-propyl) adenosine N- [(2-methyiphenyl) methyl] adenosine 2-Methyl-N- t(R)-1 -phenoxy-2-propyl] adenosine Accordingly, the present invention relates to adenosine analogues of WO 93/23418 PCT/DK93/00158 formula I HN"

RI

X N N HO HO OH wherein X is hydrogen, amino, halogen, hydroxy, lower alkoxy or lower alkyl and

R

1 is 3

R

ZR

4 wherein Y is methylene or a valence bond,

R

2 and R 5 is H or lower, straight or branched alkyi,

R

3 is H or lower alkyl, or

R

2 and R 3 can together form a cyclobutyl, cyclopentyl, cyclohexyl or phenyl ring, Z is oxygen, methylene, sulphur, sulphonyl or a valence bond,

R

4 is H, lower alkyl, aralkyl a mono or bicyclic aromatic system optionally substituted with halogen, hydroxy, haloalkyl, alkyl, alkoxy, aryloxy, acyloxy or alkylmercapto radicals, or a pharmaceutically acceptable salt thereof as these compounds have been found useful in treatment of a number of CNS-related ailments, such as cerebral ischaemia, epilepsy and pain.

Further, the compounds of formula are found to be useful agents, for lowering plasma free fatty acid (FFA) levels, as cardiovascular agents and WO 93/23418 PCT/DK93/00158 -6also have application to myocardial ischaemia.

The invention also relates to methods of preparing the above mentioned compounds. These methods comprise: Method A A compound of formula may be prepared by reacting a substance of formula wherein L represents a leaving group such as a halogen atom a chlorine or bromine atom) or a trimethylsilyloxy group, P 1

P

2 and P 3 are the same or different and represent hydrogen or a protecting group such as benzoyl-, p-toluoyl-, lower alkanoyl- acetyl-), a substituted silyl group a trimethylsilyl or t-butyldimethylsilyl group) or in the case of P 3 a triprylmethyl group, or in the case of P 1 and P 2 a 2',3'-Q-(1-methyl)ethylidene function, with a substituted amine of general formula (III) General (A)R

/R

l N N R R' xN N' N p 3 0 0 (IV)

P

2 OP1

P

2 0 0

I

R

l

HN

30 N N

HO

HO OH WO 93/23418 PCT/DK93/00158 -7giving the compound of formula (IV) as the reaction product. In cases where P 2 and P 3 are not hydrogen an additional step will be required to remove protecting groups from in cases where the groups P1, P 2 and

P

3 are for example acetyl or benzoyl, suitable conditions for deprotection include the use of methanolic ammonia, an alkali metal carbonate in methanol, or an alkali metal alkoxide in the corresponding alcohol, Where the protecting groups are for example alkylsilicon or arylsilicon derivatives, suitable methods for deprotection include, for example, treatment with tetraalkylammonium fluorides or aqueous hydrolysis in the presence of acid or base. Where the P' and P 2 groups comprise a 2',3'-Q-(1-methyl)ethylidene function or P 3 comprises triarylmethyl, suitable conditions for deprotection include, for example, hydrolysis with aqueous mineral acid.

Method B A compound of formula wherein X represents -NH 2 Q-alkyl or hydroxy, may be prepared by reacting a substance of general formula (V) S General roces (B) HN

I-I

N

N

L N X NI N 0 3 0 p 2 0 OpI p 2 0 OP X

N

HO o HO> 6 HO OH WO 93/23418 PCI'/DK93/00158 -8- [where L is a leaving group as defined in method with a nucleophile, for example, ammonia or with an anion C 1 ,-alkoxide) to afford the product In cases where P1, P 2 and P 3 are hydrogen, compound can be obtained directly. However, in cases where P 2 and P 3 are not hydrogen an additional step will be involved to remove protecting groups from examples of conditions for removal of protecting groups are given in process In some reactions involving with the anion C 1 -alkoxide, where P 2 and/or P 3 are for example acetyl- or benzoyl-, partial or full deprotection may take place. In cases where only partial deprotection has taken place, deprotection can be completed under conditions described in method Accordingly, the present invention provides a method for treating cerebral ischaemia, epilepsy and pain in human or non-human animals, which method comprises administering an effective, non-toxic amount of a compound of formula I or a pharmaceutically acceptable salt thereof, to human or non-human animals suffering from cerebral ischaemia, epilepsy or pain.

The present invention also provides the use of a compound of formula I or a pharmaceutically acceptable salt thereof in the preparation of a medicament for use in the treatment of cerebral ischaemia, epilepsy or pain.

SThe present invention further provides a pharmaceutical composition for use in the treatment of cerebral ischaemia, epilepsy or pain which comprises an effective amount of a compound of formula I of a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. Such compositions may be prepared in the manner as described below.

Various sails of compounds of formula can be prepared which can be considered physiologically acceptable. These include addition salts derived from inorganic or organic acids, for example, acetates, fumarates, glutarates, glutaconates, lactates, maleates, methanesulphonates, phosphates, VO 93/23418 PCT/DK93/00158 -9salicylates, succinates, sulphates, sulphamates, tartrates and paratoluenesulphonates. In some cases, solvates of either the free nucleosides or the acid addition salts can be isolated and these solvates may, for example, be hydrates or alcoholates.

The compounds of the invention, together with a conventional adjuvant, carrier, or diluent, and if desired in the form of a pharmaceutically-acceptable acid addition salt thereof, may be placed into the form of pharmaceutical compositions and unit dosages thereof, and in such form may be employed as solids, such as tablets of filled capsules, or liquids, such as solutions, suspensions, emulsions, elixirs, or capsules filled with the same, all for oral use, in the form of suppositories for rectal administration; or in the form of sterile injectable solutions for parenteral use (including subcutaneous .administration and infusion). Such pharmaceutical compositions and unit dosage forms thereof may comprise conventional ingredients in conventional proportions, with or without additional active compounds or principles, and such unit dosage forms may contain any suitable effective amount of the adenosine receptor agonist commensurate with the intended daily dosage range to be employed.

The compounds of this invention can thus be used for the formulation of pharmaceutical preparations, e.g. for oral and parenteral administration to mammals including humans, in accordance with conventional methods of galenic pharmacy. Conventional excipients are such pharmaceutically acceptable organic or inorganic carrier substances suitable for parenteral or enteral application which do not deleteriously react with the active compounds.

Examples of such carriers are water, salt solutions, alcohols, polyethylene glycols, polyhyroxyethoxylated castor oil, gelatine, lactose, amylose, magnesium stearate, talc, silicic acid, fatty acid monoglycerides and diglycerides, WO 93/23418 PCT/DK93/00158 pentaerythritol fatty acid esters, hydroxymethylcellulose and polyvinylpyrrolidone.

The pharmaceutical preparations can be sterilized and mixed, if desired, with auxiliary agents, emulsifiers, salt for influencing osmotic pressure, buffers and/or colouring substances and the like, which do not deleteriously react with the active compounds.

For parenteral application, particularly suitable are injectable solutions or suspensions, preferably aqueous solutions with the active compound dissolved in polyhydroxylated castor oil.

Ampoules are convenient unit dosage forms.

Tablets, dragees, or capsules having talc and/or a carbohydrate carrier or binder or the like, the carrier preferably being lactose and/or corn starch and/or potato starch, are particularly suitable for oral application. A syrup, elixir or the like can be used in cases where a sweetened vehicle can be employed.

Generally, the compounds of this invention are dispensed in unit form comprising 0.05-100 mg in a pharmaceutically acceptable carrier per unit dosage. The dosage of the compounds according to this invention is 0.1-300 mg/day, preferably 10-100 mg/day, when administered to patients, e.g. humans, as a drug.

A typical tablet which may be prepared by conventional tabletting technique3 contains: Active compound 5.0 mg Lactosum 67.8 mg PhEur, WO 93/23418 PCI'/DK93/00158 -11 Avicel® 31.4 mg Amberlite®IRP 88 1.0 mg Magnesii stearas 0.25 mg Ph.Eur.

Owing to activity against pain or convulsive disorders and prevention of neurodegeneration under conditions of anoxia/ischaemia the compounds of the invention are extremely useful in the treatment of related symptoms in mammals, when administered in an amount effective for agonist activity of compounds of the invention. The compounds of the invention may accrdingly be administered to a subject, a living animal body, including a human, in need of an adenosine receptor agonist, and if desired in the form of a pharmaceutically-acceptable acid addition salt thereof (such as the hydrobromide, hydrochloride, or sulfate, in any event prepared in the usual or conventional manner, evaporation to dryness of the free base in solution together with the acid), ordinarily concurrently, simultanously, or together with a pharmaceutically-acceptable carrier or diluent, especially and preferably in the form of a pharmaceutical composition thereof, whether by oral, rectal, or parenteral (including subcutaneous) route, in an effective amount of adenosine receptor agonist, and in any event an amount which is effective for the treatment of anoxia, traumatic injury, ischemia, migraine or other pain symptoms, epilepsy, or neurodegenerative diseases owing to their adenosine receptor agonist activity. Suitable dosage ranges are 1-200 milligrams daily, 10-100 milligrams daily, and especially 30-70 milligrams daily, depending as usual upon the exact mode of administration, form in which administered, the indication toward which the administration is directed, the subject in volved and the body weight of the subject involved, and the preference and experience of the physician or veterinarian in charge.

The preparation of compounds of the invention is further illustrated in the following examples: WO 93/23418 PCI'/DK93/00158 12- Hereinafter, TLC is thin layer chromatography, THF is tetrahydrofuran, TFA is trifluoracetic acid and mp is melting point. Where melting points are given, these are uncorrected. The structures of the compounds are confirmed by assignment of NMR spectra (from which representative peaks are quoted) and by microanalysis where appropriate. Compounds used as starting materials are either known compounds or compounds which can be prepared by methods known per se. Column chromatography was carried out on Merck silica gel 60 (Art 9385). HPLC was carried out on a Waters or Merck chromatograph with a multiwavelength detector and a reversed phase C18 column (250 x 4 mm, 5gm, 100A; eluent flow rate 1 mL/ min at 35 0 Retention times are given in minutes.

EXAMPLE 1 (Method A) 2-Chloro-N-(1-phenoxv-2-propyl)adenosine The title compound was prepared by reacting 1-phenoxy-2-propylamine (16.62 g, 0.11 mol) with 9-(2,3,5-tri-Q-acetyl-B-D-ribofuranosyl)-2,6-dichloro- 9H-purine (24.6 g, 55 mmol) in dioxan (250 ml) in the presence of triethylamine (7.23 g, 71.5 mmol) followed by deprotection of the product using a solution of sodium (0.15 g, 6.5 mmol) in methanol (250 ml). The reaction mixture was neutralized with citric acid, and treated with a mixture of ethyl acetate (300 ml) and water (200 ml). The ethyl acetate phase was separated, dried (MgSO 4 and evaporated before being purified by flash chromatography on silica gel, eluting initially with dichloromethane, and later with a mixture of dichloromethane and ethanol This provided the title 2-chloro-N-(1-phenoxy-2-propyl)adenosine (18.2 g, 76%) (a mixture of diastereoisomers) as an amorphous foam, 'H NMR (DMSO-de)5 1.31 (3H, d, -CH 3 3.53 3.59 (1H, m, 3.64 3.71 (1H, m, 3.95 (1H, q, 4.06- 4.20 (3H, 2 m, H-3' and -CH 2 4.54 (1H, m, 4.65 (1H, m,

-CHCH

3 5.07 (1H, t, 5.21, 5.50 (2H, 2d, and 5.84 (1H, d, WO 93/23418 PCT/DK93/00158 -13- 6.87 7.00 (3H, m, Ar-H), 7.23 7.32 (2H, t, Ar-H), 8.31 8.45 (2H, m, H-8 and N-H).

The corresponding maleate salt was prepared by dissolving the above 2-chloro-N-(1-phenoxy-2-propyl)adenosine (1.7 g, 3.9 mmol) in THF (10 ml), adding diethyl ether (60 ml) followed by maleic acid (0.45 g, 3.9 mmol). The residue on evaporation was treated with diethyl ether (50 ml) whereupon the maleate salt precipitated and was collected by filtration (1.15 m.p.

102-1040C.

C

23

H

26

CIN

5 0 7 requires C, 50.0; H, 4.7; N, 12.7. Found: C, 50.3; H, 4.9; N, 12.7%.

EXAMPLE 2 (Method A) 2-Chloro-N-[(R)-1-phenoxy-2-propyl1adenosine (R)-N-(tert-Butoxycarbonyl)-2-amino-1 -propanol (R)-2-Amino-1-propanol (15.0 g, 200 mmol) was dissolved in 1N sodium hydroxide (198 ml) and THF (85 ml) was introduced. The reaction mixture was cooled to 0°C and a solution of di-tert-butyl dicarbonate (52.4 g, 240 mmol) in THF (230 ml) was added dropwise over 30 min. The reaction mixture was stored at 4°C for 72 allowed to reach room temperature and filtered. The filtrate was evaporated to remove THF and the aqueous phase was extracted with ethyl acetate (2 x 200 ml). The combined extracts were dried (MgSO 4 evaporated and the crude product was dissolved in dichloromethane (100 ml) and extracted into water (5 x 200 ml). The combined aqueous extracts were evaporated in vacuo. The resultant oil crystallised whilst standing at room temperature to provide the required alcohol WO 93/23418 IPCI'/DK93/00158 -14- (15.35g, mp 59 61°C, 'H NMR (DMSO-d,)s 1.15 (3H, d, -CHCH,), 1.45 (9H, s, butyl-CH 3 3.50 (1H, dd, -CH 2 3.65 (1H, dd, -CH 2 3.70 3.80 (1H, m, CH).

(R)-N-(tert-Butoxycarbonyl)-1-phenoxy-2-propylamine (R)-N-(tert-butoxycarbonyl)-2-amino-1-propanol (10.0 g, 57 mmol), triphenylphosphine (22.5 g, 86 mmol) and phenol (5.4 g, 57 mmol) was dissolved in toluene (200 ml). Diethyl azodicarboxylate (14.9 g, 86 mmol) in toluene (100 ml) was slowly added keeping the temperature below 35°C (Mitsunobu, Synthesis, 1981, 1; Manhas, Hoffman, Lal, Bose, J. Chem. Soc. Perkin Trans 1, 1974, 461). The resulting yellow solution was stirred for 16 h at room temperature before being washed with 1N hydrochloric acid (3 x 100 ml). The organic phase was dried (MgSO 4 evaporated in vacuo, and the residual oil was purified by flash chromatography eluting with heptane/ethyl acetate giving the desired product g, 'H NMR (DMSO-de)s 1.10 (3H, d, -CH 3 1.38 (9H, s, butyl-

CH

3 3.70 3.90 (3H, m, -CH-CH 2 6.85 6.95 (3H, m, Ar-H), 7.25 (2H, t, Ar-H).

-Phenoxy-2-propylamine (R)-N-(tert-Butoxycarbonyl)-1-phenoxy-2-propylamine (8.0 g, 33 mmol) was dissolved in ethyl acetate (100 ml). A solution of hydrochloric acid in ethyl acetate (6N, 100ml) was added dropwise at room temperature. The reaction mixture was stirred at room temperature for 20h during which time a heavy precipitate was formed. The reaction mixtlure was concentrated to half the original volume before the product was collected by filtration and dried in vacuo to provide the title compound as at white solid hydrochloride (4,3 g, 69%) m.p. 186 1890C. 'H NMI (DMSO-d 6 )6 1,31 (3H, d, -CH 3 3.51 3.60 (1H, m, 4.05 (1H, dd, -CH 2 4.12 (1H, dd, -CH 2 6.95 WO 93/23418 WO 93/3418 C/DK93/OO 158 7.00 (3H, m, Ar-H), 7.32 (2H, t, Ar-H).

2-Chloro-N- [(R)l1-phenoxy-2-propyl] adenosine -phenoxy-2-propylamine (4.3 g, 23 mmol) was reacted with 9-(2,3,5-tri- O-benzoyl-B3-D-ribofuranosyl)-2,6-dichloro-9H-purine (11.2 g, 18 mmol) in dioxan (150 ml) in the presence of dilsopropylethylamine (5.3 g, 41 mmol).

The reaction mixture was stirred at room temperature for 18 h, heated at 5000 for 4h, and stirred at room temperature for 60h before being filtered and evaporated. The product (after purification by flash chromatography) was debenzoylated with methanolic ammonia to provide the title 2-chloro-N-[ -phenoxy-2-propyl] adenosine (after column chromatography) as a foarr. (4.2 g, 'H NMR (DMSO-ds 1.31 (3H, d, -OH 3 3.52 3.59 (1 H, m, 3.63 -3.72 (1 H, m, 3.92 3.99 and 4.10 4.21 (4H. 2 m, H-4' and~ -OH 2 4.52 (1 H, dd, 4.65 (1 H, m, 5.07 (1 H, t, 5.22, 5.49 (2H, 2d, 2' and 3'-OH), 5.84 (1 H, d. H-i 6.88 7.02 (3H, m, Ar-H), 7.24 7.33 (2H, dd, Ar-H), 8.32 8.45 (2H, s m, H-B and N-H).

C

19 H22CIN, 5 0, requires 0, 52.4; H, 5.1; N, 16.1. Found: C, 52.0; H, 5.2; N, 15.8%.

EXAMPLE 3 (Method A) 2-Chloro-N-r(S)..-phenoxv-2-propylladenosine 2-Chloro-Nj- -phenoxy-2-propyl) adenosine was prepared by the procedure described for Example 2, except that (S)-2-amino-1 -propanol was used in the first step, providing the opposite diastereolsomer to Example 2.

The nucleoside was obtained as a hemihydrate:

C

19 H22CIN 5 0,.0.5 H 2 0 requires C, 51.8; H, 5.2; N, 15.9. Found: C, 51.8; H, WO 93/23418 WO 93/4 18P/DK93/OO 158 16- 5.3; N, 15.6%.

EXAMPLE 4 (Method A) 2-Chloro-N-(2-phenoxvethF) adenosine The title compound was prepared by reacting 2-phenoxyethylamine hydrochloride (0.80 g, 4.6 m mol) with 9- (2,3,5-tri-O-be nzoyl B.D-rib of uranosyl)-2,6-dichloro-9H-purine (2.0 g, 3.2 mmol) in dioxan (25 ml) in the presence of triethylamine (1.0 g, 9.6 mmol) followed by deprotection of the purified product using methariolic ammonia to provide the title nucleoside (0.75 g, 60%) (following flash chromatography) as an amorphous foam, 1

H

NMVR (DMSO-dr 6 3.52 3.59 (1 H, m, H-5' 8 3.64 3.71 (1 H, m, H-5'b) 1 3.82 (2H, q, -OH 2 3.96 (1H, q, 4.14 (1H, m, 4.52 (1H, q, 5.12 (1 H. t, 5.25, 5.54 (2H, 2d, and 5.85 (1 H, d, 6.92 7.02 (3H, m, Ar-H), 7.26 -7.34 (2H, t, Ar-H), 8.46 (1 H, m, 8.56 (1 H, br t, N-H).

0 18

H

20 01N 5 0 5 .0.75 H 2 0 requires C, 49.7; H, 5.0; N, 16.1. Found: C, 49.7; H, 5.0; N, 15.7%.

EXAMPLE 5 (Method A) 2-Chloro-NR-l'(' -phenyl-2-rropvl1 adenosine The title compound wao prepared by reacting L-amphetamine (0.49 g, 3.6 rnmol) with 9-(2,3,5-tri-O-benzoyl-B3-D-ribofuranosyl)-2,6-dichloro-9H-purine (1.9 g, 3.0 mmol) in dioxan (25 ml) in the presence of d iisopropyl ethyl ami ne (0.58 g. 4.5 mmol) followed by deprotection of the purified product using methanolic ammonia. Evaporation of the reaction mixture provided a gum- WO 93/23418 WO 9/2348 1c1/DK9.'3/OO 158 17my residue which crystallized on addition of dichloromethane (10 ml), to provide the title compound (0.26 g, 38%) as a solid, m.p. 132.5 135.50C. A further sample of the title compound (0.26 g) was obtained by flash chromatography of the mother liquors. 1 H NMVR (DMSO-d 6 )6 1.22 (3H, d, -_CH3), 2.67 2.79 2.92 3.03 (2H, 2m, 3.51 3.58 (1 H, m, H-5' 0 3.62 3.68 (1 H, m, 3.94 (1 H, q, 4.12 (1 H, m, 4.41 4.54 (2H, m, H-2' and CH 3 5.06 (1 H, t, 5.22, 5.49 (2H, 2d, 2' and 3'-OH), 5.82 (1 H, d, 7.10 -7.33 (5H, m, Ar-H), 8.28 8.44 (2H, m, H-B and N-

H).

C

19 H22CIN.,O 4 .0.5 H 2 0 requires 0, 53.2; H, 5.4; N, 16.3. Found: 0, 53.3; H, 5.4; N, 16.3%.

EXAMPLE 6 (Method A) 2..Chloro-N-(1 -phenvl-3-butv)adenosine 3-Amino-i -pheriylbutane (0.67 g, 3.6 mmol) was reacted with 9-(2,3,5-tri- O-benzrjyl-B3-D-ribofuranosyl)-2,6-dichloro-9H-purine (1.9 g, 3 mmol) in dioxarn (25 ml) in the presence of diisopropylethylamine (0.58 g, 4.5 mmol), The reaction mixture was stirred at room temperature for 18 h, filtered and evaporated. The product (after purification by flash chromatography) was debenzoylated with methanolic ammonia to provide the title 2-chloro-N-(i -phenyl-3-butyl)adenosine (mixture of diastereolsomers) as a foam (0.72 g, 'H NMR (DMSO-d 6 6 1.12 (3H, d, -OH 3 3.53 3.61 (1iH, m, H-5' 0 3.64 3.72 (1 H, m, H-5ob) 1 3.93 3.99 (1 H, m, 4.11 4.17 m, H- 4.22 4.36 (1iH, m, -CH 2 4.53 (1 H, dd, 4.65 (1 H, M, -CF1 3

CH-

5.10 (1iH, t, 5.25, 5.51 (2H, 2d, 2' and 5.83 (1 H, d, 7.12 7.30 (5H, m, Ar-H), 8.28 8.37 (1 H, m, 8.40 (1iH, s, H. 8).

WO 93/23418 9323418Pcr/DK93/00158 18-

C

20

H

24 CIN 5 0 4

.H

2 0 requires 0, 53.2; H, 5.8; N, 15.5. Found: 0, 53.9; H, 5.7; N, 15.6%.

EXAMPLE 7 (Method A) N-(1 -Phenoxv-2-pjropyl)adenosine 1 -Phenoxy-2-propylamine (0.33 g, 2.18 mmol) was reacted with 6-chioropurine riboside 9-B3-D-ribofuranosyl-9H-purine) (0.5 g, 1.7 mmol) in dioxan (30 ml) in the presence of diisopropylethylamine (0.28 g, 2.2 mmol).

The reaction mixture was heated at reflux for 5 h, cooled and evaporated, The residue was purified by flash chromatography on silica gel to provide the N-(1-Phenoxy-2-propyl)adenosine (a mixture of diastereolsomers) as a foam (0.069g, 'H NMVR (DMSO-d 6 )S 1.32 (3H, d, -OH 3 3.52 3.60 (1 H, m, 3.64 3.71 (1 H, m, 3.93 3.99 and 4.1 2 4.20 (4H, 2m, m, H-3' and -OH 2 4.62 (1 H, q, 4.68 4.82 (1 H, m, -CHCH 3 5.40 (1 H, t, 51-OH), 5.20, 5.45 (2H, 2d, and 5.90 (1 H, d, 6.88 6.98 (3H, m, Ar-H), 7.23 7.31 f'2H, t, 7.85, 8.21, 8.38 (3H, 3s, H-2, H- 8 and N-H).

EXAMPLE 8 (Method A) 2-Amino-N-(l -rphenoxv-2-prorfladenosine 1 -Phenoxy-2-propylamine (2.90 g, 19.2 mmol) and 9-(2,3,5-tri-O-acetyl-13-Dribofuranosyl)-2-amino-6-chloro-9H-purine (6.94 g.,16.2 mmol) were dissolved in dioxan (50 ml) arnd triethylamine (4.5 ml, 33.2 mmol) was introduced.

After stirring the reaction mixture for 18 h at room temperature, diisopropylethylamine (2.08 g.,16.1 mmol) was added and the solution was heated at 8000 for 100 h. Following column column chromatography, a 1,3 g sample of the resultant 2',3',5'-tri-O-acetyl-2-amino-N-(1 -phenoxy-2-propyl) adenosi- WO 93/23418 TICT/DK93/00158ls 19ne was deprotected using saturated methanolic ammonia (50 ml). The reaction mixture was evaporated, and the residue dissolved in in a mixture of ethyl acetate (150 ml) and water (150 ml). The phases were separated and the ethyl acetate phase was washed with water (2 x 150 ml). The ethyl acetate phase was then extracted with pH 2 dilute hydrochloric acid, and this acidic aqueous phase was washed with ethyl acetate (2 x 100 ml), and basified with sodium bicarbonate solution before extraction with ethyl acetate (100 ml). The ethyl acetate phase was was dried (MgSO 4 and evaporated to give the title compound (0.43 g, 33%) a mixture of diastereoisomers as an amorphous foam, 'H NMR (DMSO-d 6 s) 1.28 (3H, d, -CH 3 3.49 3.57 (1H, m, 3.61 3.68 (1H, m, 3.85 3.94 and 4.07 4.15 (4H, 2m, m, H-3' and -CH 2 4.50 (1H, q, 4.68 (1H, br,

CCH

3 5.11, 5.36 (2H, 2d, and 5.40 (1H, t, 5'-OH) 5.73 (1H, d, 5.83 (1H, br, -NH 2 6.88 6.96 (3H, m, Ar-H), 7.22 7.31 (2H, t, Ar-H), 7.95 (1H, s, H-8).

C

1

,H,

4 CINsO..0.75 H 2 0 requires C, 53.1; H. 6.0; N, 19.5. Found: C, 53.0; H, N, 19.2%.

EXAMPLES 9 and 10 (Method A) Trans)-2-hydroxvcyclopentylladenosine and Trans)-2-hydroxvcyclopentvll aderosine Trans-2-hydroxycyclopentylamine (0.35 g, 3.46 mmol) (prepared by reaction of cyclopentene oxide with ammonia in a sealed vessel: see example 11) was reacted with 6-chloropurine riboside 9-A-D-ribofuranosyl-9H-purine) g, 1.7 mmol) in dioxan (30 ml) in the presence of triethylamine (0.93 g, 9 mmol). The reaction mixture was heated at 100°C for 70 h, cooled and evaporated. The resultant residue was purified by flash chromatography eluting with a mixture of ethyl acetate and methanol The fractions WO 93/23418 PCr/DK93/00158 found to contain the highest amounts of trans)-2-hydroxycyclopentyl]adenosine following HPLC examination, were combined and evaporated to a solid (0.17 Recrystallisation from methanol provided the pure Ntrans)-2-hydroxycyclopentyl]adenosine (0.11 g, 18%) mp 233-2350C.

'H NMR (DMSO-d s )6 1.43 2.12 (6H, 4m, -CH2CH 2

CH

2 3.52- 3.59 (1H, m, 3.54- 3.71 (1H, m, 3.97 (1H, q, 4.07 (1H, br, CHOH) 4.15 (1H, q, 4.61 (1H, q, 4.87, 5.21, 5.41 5.47 (4H, d 3m, OH groups), 5.89 (1H, d, 7.75 (1H, br d, 8.21 and 8.37 (H-2 and H-8).

The mother liquors from the above recrystallisation were evaporated and purified by short path chromatography on silica gel (Art. 7729) and the product recrystallised to provide trans)-2-hydroxycyclopentyl]adenosine (0.05 g, 'H NMR (DMSO-d 6 )6 1.44 2.13 (6H, 4m, -CH 2

CH

2 3.52 3.59 (1H, m, 3.54 3.71 (1H, m, 3.96 (1H, q, 4.15 (1H, q, 4.60 (1H, q, 5.20, 5.41 5.47 (3H, d m, 3' and OH), 5.88 (1H, d, 7.75 (1H, br d, 8.19 and 8.36 (H-2 and H-8).

EXAMPLE 11 (Method A) 2-Chloro-N-(cis-2-phenoxvcvclopentyl)adenosine trans-N-(tert-Butyloxycarbonyl)-2-hydroxycyclopentylamine This compound was prepared as a mixture of enantiomers by reaction of cyclopentene epoxide (8.0 g, 95.1 mmol) with a 25% aqueous ammonia solution (35 ml) in a sealed glass vessel at 110°C for 1.5 h. The reaction mixture was cooled and evaporated to half its original volume before 1N sodium hydroxide solution (95 ml) and THF (100 ml) were introduced at 0°C. A solution of di-tert-butyl dicarbonate (21.8 g, 99.6 mmol) in THF ml) was added dropwise and the reaction mixture stirred at room temperature for 18 h. The phases were separated and the aqueous phase was WO 93/23418 PCI'/DK93/00158 -21 washed with ethyl acetate (100 ml). The organic phases were combined and washed with saturated brine (100 ml), dried (MgSO 4 and evaporated.

The solid residue was recrystallised from a 10:1 mixture of heptane and ethyl acetate (55 mi) to provide an analytical sample of rans-N-(tert-butyloxycarbonyl)-2-hydroxycyclopentylamine (4.06 g, mp 103 1050C,

C

10

H

19

NO

3 requires C, 59.7; H, 9.5; N, 7.0. Found: C, 59.6; H, 9.8; N, The above trans-N-(tert-butyloxycarbonyl)-2-hydroxycyclopentylamine was converted into cis-2-phenoxy-cyclopentyl-amine by the sequence of reactions described in Example 2 phenyl ether formation by the Mitsunobu procedure resulting in inversion at the 2-position, followed by acidic hydrolysis of the BOC- group using TFA).

cis-2-Phenoxycyclopentylamine (0.75 g, 4.23 mmol) was combined with 9-(2,3,5-tri-O-benzoyl-B-D-ribofuranosyl)-2,6-dichloro-9H-purine (2,95 g, 4.7 mmol) and triethylamine (0.64 g, 6.3 mmol) in dioxan (30 ml) and stirred for h. The reaction mixture was filtered, evaporated and the residue dissolved in ethyl acetate and washed with water (2 x 50 ml). The organic phase was dried (MgSO 4 evaporated and the residue coevaporated to give cis-2',3',5'-tri-O-benzoyl-2-chloro-N-(2-phenoxycyclopentyl)adenosine (3.1 g, 90%) as an amorphous foam, which was deprotected using saturated methanolic ammonia (50 ml), After 70 h at room temperature the reaction mixture was evaporated and the residue purified by flash chromatography on silica gel, eluting with a mixture of dichloromethane and methanol The title cis-2-chloro-,. (2 phenoxycyclopentyl)adenosine (0.925 g, 53%) was obtained as an amorphous foam (a 1:1 mixture of diastereoisomers), 'H NMR (DMSO-d 6 )8 1.58 2.15 (6H, 3m, -CH 2

CH

2

CH

2 3,51 3.60 (1H, m, 3.62 3.71 (1H, m, 3.95 (1H, br q, 4.12 (1H, br q, 4,46 4.62 (2H, m, H-2' and 4.82- 4.89 (1H, m, -CH), 5.07 (1H, br t, 5.22, 5.49 (2H, 2d, and 5.83 (1H, d, WO 93/23418 PC1'/DK93/00158 -22- 6.80 6.91 (3H, m, Ar-H), 7.15 7.24 (2H, t, Ar-H), 7.99, 8.22 (1H, d m, N- 8.41, 8.45 (1H, 2s, H-8).

C

21

H

24

CIN

5 sO.0,5 H 2 0 requires C, 53,6; H, 5.4; N, 14.9. Found: C, 53.5; H, 5.3; N, 14.7%.

EXAMPLE 12 (Method A) Trans-2-chloro-N-(2-phenoxvcyclopentvl)adenosine Cis-N-(tert-butyloxycarbonyl)-2-hydroxycyclopentylamine Trans-N-(tert-butyloxycarbonyl)-2-hydroxycyclopentylamine (24.7 g, 123 mmol) (prepared as described in Example 11) was dissolved in THF (500 ml) and 4-nitrobenzoic acid (20.51 g, 123 mmol) was added, followed by triphenylphosphine (48.28 g, 184 mmol). A solution of diethylazodicarboxylate (32.06 g, 184 mmol) in THF (250 ml) was introduced dropwise. The reaction mixture was stirred for 18 h at room temperature, evaporated and purified by flash chromatography eluting with a mixture of cycohexane and ethyl acetate to provide the intermediate 4-nitrobenzoyl ester as a solid (25.5 TLC R, 0.52 [SiO 2 cyclohexane/ ethyl acetate This ester was suspended in a mixture of a mixture of methanol (180 ml) and 25% aqueous ammonia solution (20 ml) and the mixture was stirred at room temperature for 70 h before evaporation to a residue. Purification by flash chromatography eluting with a mixture of cycohexane and ethyl acetate provided fractions containing the title compound which crystallised on evaporation to afford cis-N-(tert-butyloxycarbonyl)-2-hydroxycyclopentylamine as a solid (11.0 g, mp 64 650C.

This cis-N.(tert-butyloxycarbonyl)-2-hydroxycyclopentylamine was converted WO 93/23418 WO 93/3418 C/DK93/OO 158 23 into trans-2-phenioxycyclopentylamine hydrochloride by Mitsunobu phenyl ether for~mation and deprotection the methods described in Example 2.

9-(2,3,5-Tri-O-benzoyl-i3-D-ribofuranosyl)-2,6-dichloro-9H-purine (3.0 g, 4.7 mmol) was dissolved in dioxan (30 ml) and trn-2-phenoxycyclopentylamine hydrochloride (0.95 g, 4.4 mmol) was added followed by triethylamine (0.64 g, 6.3 mmol). The reaction mixture was stirred at room temperature for 72 h and purified by flash chromatography on silica gel to provide a foam to which saturated methanolic ammonia (100 ml) was added. After 16 h at room temperature, the reaction mixture was evaporated and purified by flash chromatography to provide the title trans-2-chloro-N-(2-phenoxycyclopentyl)adlene-aine (0.70 g, 35%) as an amorphous foam (a mixture of diastereoisomers), 'H NMR (DMSO-d 6 )6 1.56 2.30 3m, -OH 2

CH

2

CH

2 3.52 3.60 (1 H, m, 3.63 3.71 (1 H, m, 3.96 (1 H, q, 4.13 (1 H, q, 4.50 -4.61 (2H, m, H-2' and 4.82 4.89 (1 H, m, -OH), 5.08 (1 H. t, 5.23, 5.49 (2H, 2d, and 5.83 (1 H, d, 6,90, 7.07 and 7.25 (5H, t,d,t, Ar-H), 8.43 (1 H, s, 8,60 (1 H, d, N-H).

021 H 24

CIN

5 0 5 0.5 H 2 0 requires C, 53.6; H, 5,4; N, 14.9. Found: 0, 53.4; H, 5.5; N, 14.5%.

EXAMPLE 13 (Method A) 2-Chloro-N- r(R) I-hydroxv-2-propyll adenosine (R)-2-Amino-l-propanol k0.23 g, 3.0 mmol), 9-(2,3,5-tri-O-benzoyl-3-D-ribofuranosyl)-2,6-dichloro-9H-purine (1.7 g, 2.7 mmol) and triethylamine (0.30 g, 3.0 mmol) were dissolved in dioxan (20 ml) and stirred for 200 h at room temperature. Following purification by column chromatography, the resultant ,5'-tri-O-benzoyl-2-chloro-N- (R)-l1-hydroxy-2-propyl] adenosine was de- WO 93/23418 WO 93/3418 CT/DK93/00158 24 protected using methanolic ammonia to provide the title 2-chloro-N~-[(R)-1hydroxy-2-propyl~adenosine as an amorphous foam (0.5 g, 'H NMVR (DMSO-d.),s 1.17 (3H, d, -OH 3 3.35 3.72 (4H, m, H-5'b and -OH 2 3.96 (1 H, q, 4.14 (1 H, m, 4.52 (1 H, dd, 5.08 (1 H, t, OH), 5.22, 5.49 (2H, 2d, 2' and 5.83 (1 H, ci, 8.0 (1KH, d, N-H) 8.40 (1 H, s H-8).

C

1 3

H

17 01N,0 5 .0.75 H 2 0 requires C, 41.9; H, 5.0; N, 18.8. Found: C, 42.1; H, 5.2; N, 15.8%.

EXAMPLE 14 (Method A)~ 2-Ohloro-N-rF(R'i-l -phenvlthio-2-propvflladenosine -N-tertbutyloxycarbonyl-1 -phenylthio-2-propylamine Thiophenol (1.5 g, 14 mmol) was dissolved in dry THIF (100 ml) and a oil dispersion of sodium hydride (0.30 g, '14 mmol) was added in portions under nitrogen. After stirring for 15 min. at room temperature, the mesylate ester of N-tert-butoxycarbonyl-2-hydroxypropylamine (3.2 g, 14 mmol) was added in three portions and the reaction mixture was heated at 7000 for 18 h. After cooling, water (30 ml) was added, the aqueous -,hase was separated and washed with dichloromethane (50 ml). The combined organic phases were dried (MgSO 4 and evaporated to give (R)-h!-tertbutyloxycarbonyl- 1 -phenylthio-2-propylamine as a fawn oil (3.2 g, TLC R 1 i 0.64 [S!O 2 heptane! ethyl acetate This -N-(tert-butoxycarboniyl)-1 -phenylthio-2-propylamine was converted into -phenylthio-2-propylamine hydrochloride by acidic hydrolysis using the method described in Example 2.

WO 93/23418PC/K3O15 PCT/DK93/00158 25 -Phenylthio-2-propylamine (0.4 g, 1.96 mmol) was reacted with 9- (2,3,5-tri-O-benzoyl-B3-D-ribofuranosyl)-2,6-dichloro-9H-purine (1.2 g, 1.9 mmol) in dioxan (15 ml) in the presence of triethylamine (0.4 g, 4 mmol).

The reaction mixture was stirred at room temperature for 72 h, heated at 500C for 24 h, cooled, filtered and evaporated. The product (after purification by flash chromatography) was debenzoylated with methanolic ammonia to provide the title 2-chloro-N-f(R)-1-phenylthio-2-propyl]adenosine (after column chromatography) as a foam (0.47 g, 'H NMR (DMSO-d.) s 1.34 (3H, d, -CH 3 3.01 (1 H, dd, 3.52 -3.60 (1 H, m, 3.62 -3.72 (1 H, m, 3.95 (1 H, q, 4.13 (1KH, m. 4.30 4.45 (1 H, m, 4.53 (1 H, m, 5.09, 5.22, 5.50 (3H, 3 br, 3'and 5.84 (1KH, d, 7.19 (1 H, t, Ar-H), 7.30 t, Ar-H), 7.45 (2H, d, Ar-H), 8.29 8.45 (2K, s m, H-8 and N-H).

C,,H22CIN 5 O4S requires C, 50.5; H, 4.9; N, 15., Found: C, 50.6; H, 5.1; N, 15.2%.

EXAMPLE 15 (Method A) (R)-2-Chloro-N-r1-(4-fluorophenoxv)-2-propvlladenosine (R)-1-(4-fluorophenoxy)-2-propylamine (0.29 g, 1.4 mmol) (prepared from 4fluorophenol by the method desrcribed in example 2) was reacted with 9-(2',3',5'-tri-.Q-benzoyl-B-D-ribofuranosyl)-2,6-dichloro-9H-purine (0.89 g, 1.4 mmol) in dioxan (30 ml) in the presence of triethylamine (0.42 g, 3 mmol).

The reaction mixture was stirred at room temperature for 18 h, and heated at 6000 for 4 h. The reaction mixture was filtered and evaporated to a residue which was purified by flash chromatography. The resultant 2',3',5'-tri-Obenzoyl-2-chloro-Nj-[(R)-1 -(4-fluorophenoxy)-2-propylj adenosine was deprotected using methanolic ammonia to provide the title 2-chloro-N-[(R)-1 fluorophenoxy)-2-propyl]adenosine (0.21 g, 40%) (after column chromato- WO 93/23418 WO 93/3418 Cr/DK93/0O1 58 26 graphy), mp 172 17300; 'H NMVR (DMSO-d 6 6 1.29 (3H, dl, -OH 3 3.52 3.60 (1 H, m, 3.64 3.72 (1 H, m, 3.92 4.00 m, H-4' and 4.05 4.20 (2H, m, H-3' and 4.53 (1 H, m, 4.65 (1 H, m,

CH

3 5.08, 5.24, 5.50 3 br, 3' and 5.86 (1 H, d, 6.89 7.15 (4H, 2 m, Ar-H), 8.30 8.46 m, H-8 and N-H).

C

19

H

21 C1FN,,0 5 requires 0, 49.8; H, 4.7; N, 15.3. Found: 0, 49.4; H, 4.7; N, 14.9%.

EXAMPLE 16 (Method A) 2-Chloro-N- F(R)-2-phenoxv-1 -propyll adenosine (R)-2-Phenoxy-1-propylamine (0.6 g, 2.9 mmol) (prepared by the method described in example 2) was reacted with 9-(2,3,5-tri-O-benzoyl-B3-D-ribofuranosyl)-2,6-dichloro-9H-purine (1.5 g, 2.4 mmol) in dioxan (20 ml) in the presence of triethylamine (0.5 g, 5.3 mmol). The reaction mixture was stirred at room temperature for 72 h before being filtered and evaporated. The product, following purification by flash chromatography, was treated with saturated methanolic ammonia (30 ml) for 18 h and evaporated to provide a solid residue. This solid was washed thoroughly with dichloromethane to provide the title 2-chloro-IA-[(R)-2-phenoxy--propyl]adenosine (0.7 g, mp 175 17700, 1 H NMVR (DMSO-d 6 6 1.39 (3H, d, -OH 3 3.56 (1 H, ABX, 3.68 (1 H, m, 3.33 -3.40 (1 H, m, 3.83 3.92 (1 H, m, -C- 3.96 (1 H, q, 4.14 (1 H, m, 4.53 (1 H, dd, 4.70 (1 H, q, 5.08, 5.34, 5.50 3 br, 3' and 5.85 (1H, d, 6.90 (1 H. t, Ar-H), 7.11 (2H, d, Ar-H), 7.28 t, Ar-H), 8.45 (1 H, s, 8.63 (1 H, t, N

C

19 H,2CIN 5

O

5 requires C, 52.4; H, 5.1 N, 16.1. Found: C, 52.5; H, 5.1; N, 15,9%, WO 93/23418 WO 93/3418 CT/DK93/OO 158 27- EXAMPLE 17 (Method A) 2-Ohloro-N- r2-(rhenylmethoxy) ethyl] adenosine The title compound was prepared by reacting 2-(phenylmethoxy)ethylamine hydrochloride (0.51 g, 2.7 mmol) with 9- (2,3,5-tri-O--benzoyl -8-D-ri bofu ran osyl)-2,6-dichloro-9H-purine (1.43 g, 2.25 mmol), followed by debenzoylation of the purified product using methanolic ammonia to provide the title 2chloro-N-[2-(phenylmethoxy) ethyl] adenosine (0.38 g, 44%) (after column chromatography) as a solid, mp 115 -124oC, 'H NMVR (DMSO-d 6 )S 3.50 3.58 (1 H, m, 3.60 3.70 (4H, m, H-5'b, -CH 2 and 3.95 (1 H, q, 4.04 4.16 (2H, m, H-3' and 4.52 (1H, br s, H-2' and -OH 2 5.07 (1 H, t, 5.21, 5.50 (2H, 2d, 2'-and 5.84 (1 H, d, 7.22 7.36 (5H, m, Ar-H), 8.25 8.40 (2H, m, H-8 and N-H).

0, 9 H2201N 5

O

5 0.1 H 2 0 requires 0, 52.1 5.1 N, 16.0. Found: C, 51.8; H, 5.3; N, 15.6%.

EXAMPLE 18 (Method A) 2-Fluoro-N-r(RY 1 -rhenoxv-2-rropylladenosine 9-(2,3,5-Tri-O-acetyl-j3-D-ribofuranosyl)-6-chloro-2-fluoro-9H-purine (1 .03 g, 2.38 mmol) POT Publication No. WO 93/08206, -phenoxy-2-propylamine (0.36 g, 2.38 mmol) and triethylamine (0.29 g, 0.28 mmol) in dioxan ml) were stirred at room temperature for 18 h. The reaction mixture was filtered and evaporated to a residue which was purified by flash chromatography. The resultant 2',3',5'-tri-O-acetyl-2-fluoro-N-f (R)-l1-phenoxy-2-propyijadenosine was deprotected using methanolic ammonia to provide the title 2-fluoro-N-[(R)-1-phenoxy-2-propyl]adenosine (0.28 g, 23%) (after column chromatography), mp 148 150'0; 'H NMVR (DMSO-d 6 )S 1.33 (3H, d, WO 93/23418 WO 9323418PCT/DK93/OO 158 28 3.59 (1IH, m, 3.63 -3.71 (1H, m, 3.92 -3.99 (2H, m, H-4' and 4.10 4.18 (2H, m, H-3' and 4.51 (1 H, q, 4.61 (1 H, m,

CH

3 5.06 (1 H, t, 5.22, 5.48 (2H, 2d, 2' and 5.82 (1 H, d, 6.89 6.97 (3H, m, Ar-H), 7.25 7.30 t, Ar-H), 8.39 (1 H, s, H-8), 8.49 (1H, d, N-H).

C

19 H22FN, 5

O

5 requires C, 54.4; H, 5.3; N, 16.7. Found: C, 54.7; H, 5.5; N, 16.4%.

Example 19 (Method B) 2-Methoxv-N- F(R)-1 -rhenoxv-2-Dropvll adenosine 2-Methoxy-N- -phenoxy-2-projpyl) adenosine was prepared by reacting 2-chloro-N-((R) I-phenoxy-2-propyl) adenosine (Example 2) (0.30 g, 0.69 mmol) with a mixture of sodium hydroxide (0.32 g, 8.0 mmol) and methanol ml) in a sealed vessel at 80 900C for 4h. The cooled reaction mixture was neutralised with concentrated hydrochloric acid and evaporated to dryness. Water (30 ml) was added and the mixture was extracted with dichloromethane (2 x 30 ml). The combined extracts were dried (MgSO 4 and coevaporated with dichloromethane (30 ml), giving the title compound as a foam (0.19 g, 'H NMVR (DMSO-d.) 6 1.32 (3H, d, -CHCH.

3 3.55 (1 H, m, H-5j 0 3.65 (1 H, m, H5'b), 3.72 (3H, s, -OH 3 3.91 3.99 and 4.10 4.20 (4H, 2 m, H-4' and -OH 2 4.51 (1 H, dd, 4.67 (1 H, m, -CHCH 3 5.84 (11-1, d, 6.89 6.98 m, 7.26 dd, Ar-H) 8.12 (1H-, br, 8.46 1H, s, H-8).

WO 93/23418 WO 93/23418 CT/DK193/00158 29 EXAMPLE 20 (Method A) N-(2-Methoxvethyl)adenosine The title compound was prepared by the procedure described in example 7 by reacting 2-methoxyethylamine hvdrochloride (0.27 g, 3.6 mmol) with 6-chioropurine riboside 9-3-D-ribofuranosyl-6-chloro-9H-purine) (1.0 g, mmol) in dioxan (30 ml) at room temperature for 72 h with triethylamine (1.04 ml, 7.5 mmol) present. The reaction mixture was filtered and evaporated and the resultant residue was recrystallised from methanol (100 ml) to provide the title compound (0.80 g, 82%) as a solid, mp 151 -1520C, 'H NMVR (DMSO-d 6 )6 3.26 (3H, s, -OH 3 3.50 3.58 (3H, m, H-5', 0 and -OH 2 3.60 3.70 (3H, m, H-5'b and -OH 2 3.96 (1 H, q, 4.14 (1 H, dd, 4.60 (1 H, dd, 5.20, 5.45 (2H, 2d, 2'-and 5.42 (1 H, t, 5.87 (1 H, d, 7.80 (1 H, br s, -NH) 8.22, 8.35 (2H, 2s, H-2 and H-B) C1 3 H1,N,0 5 requires 0, 48.0 H, 5.9 N, 21.5. Found: 0, 47,8; H, 5.9; N, 21.3%.

EXAMPLE 21 (Method A) 2-Chloro-N-r(2-methoxvohenvlmethyl1 adenosine The title compound was prepared by reacting (2-methoxyphenyl)methylamine (0.55 g, 4.0 mmol) with 9-(2,3,5-tri-O-benzoyl-13-D-ribofuranosyl)-2,6-dichloro-9H-purine (1.01 g, 1.6 mmol), followed by debenzoylation of the purified product using methano~c ammonia to provide the title 2-chloro-N-[(2-methoxyphenyl)mnethyl~adenosine (0.31 g, 45%) (after column chromatography) as a solid, mp 116 1190C, 'H NMVR (DMSO-d6) 6 3.51 3.60 (1 H, m, H-5' 0 3.61 3.70 (1 H, m, 3.95 (1 H, q. 4.13 (1 H, m, 4.52 (1 H. q, 5.06 (1 H, t, 5.22, 5.50 (2H, 2d, 2'-and 5.85 (1 H, d, WO 93/23418 WO 93/3418 C'/DK93/OO 158 30 6.83 7.25 (4H, 2t, 2d, Ar-H), 8.43 (1 H, s, 8.72 (1 H, t, N-H).

EXAMPLE 22 (Method A) 2-Chloro-N-[(R)-3-methvl-1 -phenoxv-2-buvladenosine (R)-3-methyl-1-phenoxy-2-butylamine (0.6 g, 2.8 mmol) was reacted with 9- (2,3,5-tri-O-benzoyl-B- D-ri bofu ran osyl) -2,6-d ich loro-9 H-pu ri ne (1 .4 g, 2.2 mmol) in dioxan (20 ml) in the presence of triethylamine (0.5 g, 5.0 mmol).

The reaction mixture was stirred at room temperature for 40 h before being filtered and evaporated. The product (after purification by flash chromatography) was debenzoylated with methanolic ammonia to provide the product (after column chromatography) as a foam which solidified on coevaporation with dichloromethane. 2-Chloro-N- [(R)-3-methyl-1 -phenoxy-2-butyl] adenosine (0.46 g. 44%) was obtained as a white solid, mp 95 10000,' 'H NMR (DMVSO-dr) s 0.95, 0.98 (6H, 2d, 2 x -OH 3 2.10 (1 H, m, -CH(0H 3 2 3.53 3.60 (1 H, m, 3.63 -3.71 (1 H, m, 5 3.95 (1 H, q, 4.07 4.23 (3H, m, H-3' and -OH 2 4.96 (1lH, m, 4.56 (1 H, q, 5.08 (1H, t. 5.23, 5.49 2d, 2' and 5.84 (1H, d, 6.87 6.97 (3H, m, Ar-H), 7.24 -7.31 (2H, d~d, Ar-H), 8.36 (1 H, d, 8.40 (I1H, s, H-8), 0 21

H

2 6 CIN,0,.0.5 H 2 0 requires C, 53.3; H, 5.8; N, 14.8. Found: C, 53.4; H, 5.7; N, 14.8%.

EXAMPLE 23 (Method A) 2-Chloro-W- R-1 -(2-(2-propvloxv) phenox )-2-propl1 adenosine 1-(2-(2-Propyloxy) phenoxy)-2-propylamine (prepared from 2-(2-propyloxy)phenol by the procedure described in example 2) (0.54 g, 2.2 mmol) WO 93/23418 WO 93/23418 PG/DK93/OO 158 31 was reacted with 9-(2,3,5-tri-O-acetyl-B3-D-ribofuranosyl)-2,6-dichloro-9Hpurine (2.0 g, 4.5 mmol) in dioxan (30 ml) in the presence of triethylamine (2.19 g, 22 mmol). The reaction mixture was stirred at room temperature for 18 h before being filtered and evaporated. The product (after purification by flash chromatography) was debenzoylated with methanolic ammonia to provide the 2-chloro-N-I(R)-1 -(2-(2-propyloxy)phenoxy)-2-propyl~adenosine (after column chromatography) as a foam (0.47 g, 'H NMVR (DMSO-d 6 1.04, 1.06 (6H, 2d, 2 x -OH 3 1.31 (3H, d, -OH 3 3.53 3.60 (1 H, m, H-5' 8 3.64 3.71 (1 H, m. H-Sib), 3.95 (1 H, q, 3.98 4.15 (3H, 2m, H-3' and -CH 2 4.35 (1 H, p, 4.51 (1 H, q, 4.72 (1 H, m, 5.08 (1 H, t, OH), 5.22, 5.48 (2K, 2d, 2' and 5.85 (1KH, d, 6.82 7.08 m, Ar-H), 8.32 (1KH, d, 8.41 (1KH, s H-8).

C22H 28

CIN

5 0 6 .1.0 H 2 0 requires C, 51.6; H, 5.9; N, 13.7. Found: C, 52.0; H, 5.8; N, 13.3%.

EXAMPLE 24 (Method A) 2-Chloro-N-[(R)-1-phenvlsulphonvl-2-rpropvlladenosine (R)-1-Phenylsulphonyl-2-propylamine (0.4 g. 1.7 mmol) was reacted with 9- (2,3,5-tri-O-benzoyl-B3-D-ribofuranosyl)-2,6-dichloro-9H-purine (1.7 g, mmol) in dioxan (20 ml) in the presence of triethylamine (0.4 g, 4.3 rmcl).

The reaction mixture was stirred at room temperature for 48 h, heated at 6000 for 4 h, cooled, filtered and evaporated. The product (after purification by flash chromatography) was debenzoylated with methanolic ammonia to provide the title 2-chloro-N- -1-phenylsulphonyl-2-propyl] adenosine (after column chromatography) as a foam (0.2 g, 'H NMR (DMSO-d,)S 1.24 (3H, d, -CH 3 3.45 (1KH, dci, 3.53 3.61 (2K, m, and 3.64 3.71 (1KH, m, 3.86 (1KH, dd, 3.97 (1KH, q, 4.14 (1KR WO 93/23418 WO 93/23418PCI',D K93/OO 158 32 4.53 (1HF, m, 5.09 (1iH, t, 5.23, 5.50 (2H, 2 d, 2' and 5.83 (1 H, d, 7.45 -7.82 (5H, m, Ar-H), 8.21 (1 H, s, 8.38 (1 H, s, H-8).

EXAMPLE 25 (Method A) N-f (2-methvhphenvl) methvll adenosine The title compound was prepared by react!.Ig (2-methylphenyl)methylamine (1.51 g, 12.5 mmol) with 6-chioropurine riboside (2.87 g, 10.0 mmol) in dioxan (100 ml) in the presence of diisopropylethylamnine (1.94 g, 15.0 mmol). The reaction mixture was heated at 6000 for 6 h, cooled, filtered and evaporated. The residue was purified by flash chromatography, eluting initially with dichloromethane, and later increasing polarity to dichloromethane/ ethanol to provide the product (2.6 g, 70%) as a solid which was recrystallised from methanol to give N-[(2-methylphenyl)methyl]adenosine as white crystals (1.75 g, mp 161.5 163.50C, 'H NMVR (DMSO-d 6

)S

2.35 (3H, s, -OHs), 3.53 3.60 (1 H, m, 3.65 3.72 (1 H, m, 3.98 (1 H, q, 4.16 (1 H, m, 4.64 (1 H, q, 5.41 (1 H, t, 5.21, 5.48 (2H, 2d, 2'-and 5.92 (1 H, d, 7.06 -7.24 (4H, m, Ar-H), 8.20 and 8.40 (3H, s and br s, H-2, H-B and N-H).

EXAMPLE 26 (Method A) 2-Methvl-N-r(R)-l -phenoxv-2-propvlladenosine (R)-l-phenoxy-2-propylai nine (0.56 g, 3 mmol) was reacted with 9-(2,3,5-tri- 0-acetyl-B-D-ribofuranosyl)-6- :hloro-2-methyl-9H-purine (0.43 g, 1 mmol) (prepared from 2-methylinosine (Journal of Organic Chemistry, 1967, 3, 3258 3260) by standard acylation and chloriration steps] in dioxan (20 ml) in the presence of triethylamine (0.41 g, 4 mmcl). The reaction mixture was WO 93/23418 WO 9/23 18PCr/DK93/00158 33 heated at 5000 for 70h, and at 9000 for 3 h. before being filtered and evaporated. The product (after purification by flash chromatography) was debenzoylated with methanolic ammonia to provide the title 2-methyl-N-[(R)- I-phenoxy-2-propyl]adenosine (after column chromatography) as a foam (0.21 g, 'H NMR (DMSO-d,)6 1.30 (3H, d, -OH 3 2.43 (3H, s, -OH 3 3.53 3.60 (1 H, m, 3.66 3.73 (1 H, m, 3.94 (1 H, dd, 3.99 (1 H, q, 4.12 4.22 (2H, rm, H-3'and 4.54 (1 H, dd, 4.76 (1 H, m, -CH 3 CII-_) 5.20, 5.52 (2H, 2d, 2' and 5.73 (1 H, t, OH), 5.87 (1 H. d, 6.90 7.31 (5H, t, m, t, Ar-H), 7.74 (1 H, br d, N-H), 8.28 (1 H, s, H-8).

0 20

H

25 01N 5 0 5 .0.33 H 2 0 requires C, 57.0; H, 6.1; N, 16.6. Found: C, 57.0; H, 6.2; N, 16.8%.

WO 93/23418 SPCT/DK93/00158 -34- Evaluation of the compounds.

Methods for assessing adenosine receptor binding in vitro have been reviewed [Adenosine Receptors, (Cooper, D.M.F. and Londos, eds.) Alan R.

Liss, Inc., New York, 1988, 43-62].

Evaluation of these compounds in established animal models has indicated that the compounds according to the invention possess desirable central nervous system properties. For example, they act as anticonvulsant agents, are effective in animal models of pain, and show cerebroprotective effects in laboratory test animals subjected to simulated cerebral ischaemia. In addition, the compounds may have efficacy as neuroprotective agents in cases of cerebral oedema and traumatic head injury.

Evaluation of in vitro binding to adenosine Al and A2 receptors The affinity of the known and novel compounds described in this invention for the adenosine Al receptor has been determined essentially as described in the literature using 3 H]-B-PIA as a radioligand (Naunyn-Schmiedeberg's Archives of Pharmacology, 1980, 313, 179-187). Affinity for the A2 receptor was measured using the radioligand 3 H]-CGS 21680 (European Journal of Pharmacology, 1989, 168, 243-246), and the values for representative compounds are given in table I below. In vitro receptor binding values obtained for the reference standard adenosine agonists CPA [N-(cyclopentyl)adenosine] and R-PIA [N-(1-phenyl-2-propyl)adenosin6]) are included for comparison.

WO 93/23418 PCT/DK93/00158 Method description DMCM INDUCED SEIZURES IN MICE In this model, seizures are induced by i.p. (intraperitoneal) dosing of methyl 6,7-dimethoxy-4-ethyl-i3-carboline-3-carboxylate DMCM at 15 mg/kg.

DMCM is an inverse agonist to the benzodiazepine receptor, presumably producing seizures by decreasing the potency of inhibition of the GABA receptor/benzodiazepine receptor/chloride ionophore complex.

mg/kg of DMCM dissolved in 0.02 N HCI (1 mg/ml) is administered i.p. In a volume of 300 Al to male NMRI mice weighing 20 2 g. This induces two different responses: a) some animals manifest a brief loss of righting reflexes or take up an upright position in which they have a mild short clonus of the upper extremities, b) other animals manifest intense clonic and tonic convulsions of all extremities often followed by death. DMCM is administered 30 min after an intraperitoneal injection of a test compound.

Latency time for the presence of intense clonic and tonic convulsions and death is noted until 15 min after administration of DMCM. At least 5 doses of each test compound are tested with 8 mice per dose.

An anticonvulsive EDso value is determined as the dose (mg/kg) protecting 50% of the animals against clonic convulsions; some representative values are shown in table II.

The above method is a described in Petersen, Eur, J. Pharmacol, 94, 117-124, 1983; Petersen, EN., Eur. J, Pharmacol. 195, 261-265, 1991.

WO 93/23418 PCr/DK93/00158 -36- Blood pre'qtre in anaesthetised rats Test compounds are generally dissolved in DMSO and diluted in 5% chremophore/saline before being dosed to nembutal anaesthetised 200 g female Sprague Dawley rats which have not been starved or fasted. The rats are breathing spontaneously; blood pressure (BP) and heart rate (HR) is measured 5 minutes after a bolus i.v. injection. Each measurement is repeated twice. Results for representative compounds are shown in table II.

Neuroprotective effect: Gerbil BCAO ischemia model.

Transient global ischaemia was produced in Mongolian gerbils (60-70 g, males) anaesthetized with 2% halothane in 70% nitrous oxide and 30% oxygen. The common carotid arteries were occluded for 5 min, and the animals were allowed to recover for 4 days, The animals were reanaesthetized, decapitated and the brains quickly removed and frozen in powdered dry ice.

Coronal sections (20 pm) were taken through the brain at the level of the hippocampus and stained with cresyl violet and hematoxylineosin. The brain sections were rated for neuronal damage in the hippocampus CA1 region using a scale from 0 (undamaged) to 3 (total damage of CA1). The body temperature of all the animals was maintained at 37°C throughout the surgery and the animals were placed in warmed boxes during the recovery period. Each experiment consisted of a drug and a vehicle control group (n 10-15). Test compounds were administered 30 min, after reperfusion.

WO 93/23418 PCT/DK93/00158 -37- TABLE I In Vitro evaluation of the compounds Adenosine agonist Al receptor A2 receptor Ratio tested binding binding A2/A1

(K

i nM) (K 1 nM) 43 1157 27 18 318 18 100 7413 74 166 3095 19 4 123 31 36 802 22 18 791 44 123 2188 18 3.3 3270 991 3.1 1320 426 (11) 35 397 11 (12) 6 383 64 (13) 7 2241 320 (14) 15 893 19 540 28 (17) 340 7776 23 (18) 8 310 39 (19) 88 2332 27 77 2432 32 (26) 69 1200 17 CPA 1.6 173 108 (R)-PIA 2,0 134 67 11 U01THMA A(.0612 170~ 17/0196 -38- TABLE 11 Pharm, ci'k11gical evaluation of the comp~ounds p..

p.

Compound (Example No.) (1) (2) (4) (6) (8) (10) (11) (12) (14) (15) (19) (26) DMCM seizures

ED

50 (m-g/kg) i.p.

HR

3.4 3.9 13.3 3.8 24.9 0.1 2.0 6.7 9,9 13.6 6.7 0.5 1.9 fall in BP 0. 1 mg/kg i. v.

0 8 0 16 22 Throughout this specification and the claivns which follow, unless the context requires otherwise, the word "comprise", or variations such as "comprises" or "comprising", wvill be understood to imply the inclusion of a stated integer or group of integers but not thle exclusion of any other integer or group of integers.

Claims (6)

1. A method for the treatment or prophylaxis of treating myocardial or cerebral ischaemia, or epilepsy which comprises administering to a patient in need of such treatnment a therapeutically effective amount of a compound of formula I 0 0* 0* 000 wherein X is hydrogen, amino, halogen, hydroxy, lower alkoxy or lower alkyl and RI is 3 204 wherein Y is methylene or a valence bond, R 2 and R 5 is H or lower, straight or branched alkyl, R 3 is IH or lower alkyl, or R 2 and R 3 can together form a cyclobutyl, cyclopentyl, cyclohexyl or phienyl ring, Z is oxygen, methylene, sulphur, suiphonyl or a valence bond, W 4 is H, lowver alkyl, aralkyl, a mono or bicyclie aromatic system optionally substituted with halogen, hydroxy, haloalkyl, alkyl, alkoxy, aryloxy, acyloxy or alkyltnercapto radicals, or a pharmaceutically acceptable salt thereof. 11:011101'I(10,10612-93,179 -27/96

2. A method according to claim 1, wherein, in the compound of formula 1, X is halogen, Z is oxygen or sulphur and W 4 is phenyl, optionally substituted with fluoro.

3. A method according to claim 1, wherein the compound of formula I is selected from 2-chloro-N-[(R)-l1-phenoxy-2-propyljadenosine, 2-chloro-N-(2-phenoxyethyl)adenosine, 2-chloro-N-(1 -phenyl-3-butyl)adenosine, 2-chloro-N-(1 -phenoxy-2-propyl)adenosine, 10 .ooN(i-2peoyylpetlaeoie

102-chloro-N-(tris-2-phenoxycyclopentyl)adenosine, 2-chloro-N-(R)n--hdoxycycropeyladenosine, 2-chloro-N-[(R)-1-hylox-2-propyladenosine, 2-chloro-N-[(R).1-4fupheni -2-propyladenosine,

152-chloro-N-[(R)1 -(-fluoroheo)--propyl]adenosine, OV, ~2-cloro-N-[2-(phienylmnethoxy)etliyl]adenosine, V, 2-fluoro-N-[(R)-1-phenoxy-2-propyl] adenosine, 2-methoxy-N-[(R)- 1-phenoxy-2-propylladenosine, 5: N-(2-methoxyethyl)adenosine, 20 2-chloro-N-[(R)-3-methyl-1 -phenoxy-2-butylladenosine, 2-cliloro-N-[(R)-l1-(2-(2-propyloxy)phenoxy)-2-propyl] adenosine, 2-cliloro-N-[(R)-1 -phenlylsulplionyl-2-propyljadenosine, N-[(2-methiylphenyl)metliyl]adenosine or 2-methyl-N-[(R)-1 -plhenoxy-2-propyl]adenosine, or pharmaceutically acceptable salts thereof. J:II1T1C46293,179 27/6196 -41- 4. A compound selected from 2-chloro-N-[XR)-i-phenoxy-2-propyljadenosine, 2-chloro-N-(2-phenoxyethyl)adenosine, 2-chloro-N-(1-phenyl-3-butyl)adenosine, 2-chloro-N-(1-phenoxy-2-propyi,)adenosine, 2-chloro-N-(cis-2-phenoxycyclopentyl)adenosine, 2-chioro-N-(trans-2-phenoxycyclopentyl)adenosine, 2-chloro-N-[(R)-1 -hydroxy-2-propylljadenosine, 10 2-chloro-N-II(R)-1-phenylthio-2-propylladenosine, 2-chloro-N-[( R) 1-(4-fliuorophenoxy)-2-propyllladenosine, 2-chloro-N-[(R)-2-phenoxy-l1-propyljadenosine, 2-chloro-N-[2-(phenylmethoxy)etliyl]adenosine, A 2-fluoro-N-[(R)- I -phenoxy-2-propyl] adenosine, 15 2-methoxy-N-[(R)-1-phenoxy-2-propylladenosinie, N-(2-methoxyethyl)adenosine, ***2-cliloro-N-[(R)-3-methiyl-1-phenioxy-2-butyl]adenosine, 2-chloro-N-[(R,)-1 -(2-(2-propyloxy)phenoxy)-2-propyl] adenosine, 2-chiloro-N-[(R)-1 -phenylsulphonyl-2-propylladenosine, 20 N-[(2-methylphenyl)methyl]adenosine or 2-iniethyl-N-[(R)- 1-phenoxy-2-propyllladenosine or pharmaceutically acceptable salts thereof. A pharmaceutical composition comprising as active component a compound according to claim 4 and a pharmaceutically acceptable carrier. 6. A pharmaceutical composition according to claim 5 in the form of an oral dosage unit containing about 1-200 mg of the active compound. 0 b 4 I':101(1VITC'IMO61293. 179- 27/6/96 -42- 7. A method for the treatment or prophylaxis of myocardial or cerebral ischaemia or epilepsy, substantially as hereinbefore described with reference to the Examples. 8. Compounds of Formula I methods for their preparation or pharmaceutical compositions or methods of treatment involving them substantially as hereinbefore described with reference to the Examples. DATED this 27th day of June 1996 10 Novo Nordisk A/S by DAVIES COLLISON CAVE Patent Attorneys for the Applicant 0 000000 0 0* 0 000 0 *00 0 0 0 ~i 6 ,q 1 INTERNATIONAL SEAiiCH REPORT International application No, PCT/DK 93/00158 I A. CLASSIFICATViN F SUBJECT MATTER IPC5: C07H 19/16 C07H 19/167 A61K 31/70 According to International Patent Classification or to both national classification and IPC B. FIELDS SEARCHED Minimum documentation searched (classification system followed by classification symbols) C07H, A61K Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched SE,DK,FI,NO classes as above Electronic data base consulted during the international search (name of data base and, where pt acticable, search terms used) CA, MEDLINE C. DOCUMENTS CONSIDERED TO BE RELEVANT Category* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No. X US, A, 4791103 (TRIVEDI ET AL), 13 December 1988 1-8 (13.12.88), the claims X US, A, 3551409 (WOLFGANG KAMPE ET AL), 1-8 29 December 1970 (29.12.70) X US, A, 3502649 (MAX THIEL ET AL), 24 March 1970 1-8 (24.03.70), see part examples 1-3,9,15-18 X EP, A2, 0322242 (GLAXO GROUP LIMITED), 1-8 28 June 1989 (28.06.89), see part page 3 and the exemples Further documents are listed in the continuation of Box C. See patent family annex. Special categones of cited documetr T' later document published after the international filing date or pnonty A date and not in conflict with the application but cited to understand A' document defining the eneral state of the an whch s not considered the pnnciple or theory underlying the invention to be of particular relevance E* erier document but published on or after the intenational iling date document of particular relevance: the claimed inv'ttbon cannot be considered novel or cannot be considered to involve inventive L" docunent ibch may throw doubts on pnonty claom(s) or whlch s step wh the dcument S an alone cited to establish the publication date of another ctUaon or other um t i alo special reason (as specified) document of particular relevance: the claimed invention cannot be document refernng to an oral duclosure, use, exhibition or other considered to involve an inventive step when the document is means combined with one or more other such document, such combination document publisbed pnor to the internatonal filing date but later than being obvious to a person skilled in the art the pnonty date claimed document member of the same patent family Date of the actual completion of the international search Date of mailing of the international search report 23 August 1993 '25 -08- 1993 \ame and mailing address of the ISA, Authorized oficer Swedish Patent Office Box 5055. S-102 42 STOCKHOLM Eva Johansson Facsimile No. +4 6 8 666 02 86 Telephone No. +46 8 782 25 00 Form PCT/ISAi210 (second sheet) (July 1992) INTERNATIONAL SEARCH REPORT International application No. PCT/DK 93/00158 C (Continuation). DOCUMENTS CONSIDERED TO BE RELEVANT Category* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No. X EP, A2, 0253962 (HOECHST JAPAN LIMITED), 1-8 27 January 1988 (27.01.88), page 5 page 61 X EP, A2, 0152944 (BOEHRINGER MANNHEIM GMBH), 1-8 28 August 1985 (28.08.85), see part table 3 compounds 5,40,42,46,61 and 67 X DE, A, 2052596 (BOEHRINGER MANNHEIM GMBH), 4 May 1-8 1972 (04.05.72) X DE, B, 1670077 (BOEHRINGER MANNHEIM GMBH), 1-8 14 August 1974 (14.08.74), see part the examples X J. Med. Chem., Volume 28, 1985, Shozo Kusachi et 1-8 al, "Dog Coronary Artery Adenosine Receptor: Structure of the N6-Alkyl Subregion" page 1636 page 1643 X Chemical Abstracts, Volume 96, No 15, 1-8 12 April 1982 (12.04.82), (Columbus, Ohio, USA), Dunwiddie, Thomas V et al, "Sedative and anticonvulsant effects of adenosine analogs in mouse and rat", page 135, THE ABSTRACT No 116488b, J, Pharmacol. Exp. Ther. 1982, 220 70-76 X Chemical Abstracts, Volume 76, No 23, 5 June 1972 1-8 (05.06.72), (Columbus, Ohio, USA), Fleysher, M. H., "N6-Substituted adenosines. Synthesis, Biological activity, and some structure-activity relations", page 11, THE ABSTRACT No 135537v, J. Med. Chem.,

187-191 Form PCTIISA/210 (ConunuAuon of second thee) (July 1992) INTERNATIONAL SEARCH REPORT International application No. PCT/DK 93/00158 C (Continuation). DOCUMENTS CONSIDERED TO BE RELEVANT Category* Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No. X Chemical Abstracts, Volume 72, No 1, 1-8 January 1970 (05.01.70), (Columbus, Ohio, USA), Fleysher, Maurice H et al, "Synthesis and biological activity of some new N6-substituted purine nucleosides", page 349, THE ABSTRACT No 3706g, J. Med. Chem. 1969, 12 1056-1061 X Dialog Information Services, Medline, 1-8 Dialog accession no. 91211907, Karlsten R et al: "The antinociceptive effect of inttrathecally administrered adenosine analogs in mice correlates with the affinity for the Al-adenosine receptor", Neurosci Lett, (1991 Jan 2) 121 267-70 X Chemical Abstracts, Volume 70, No 19, 1-8 28 April 1969 (28.04.69), (Columbus, Ohio, USA), Kampe, Wolfgang et al, "Adenosines", page 394, THE ABSTRACT No 88212z, S. African 1968, A WO, Al, 8504882 (NELSON RESEARCH AND DEVELOPMENT 1-8 COMPANY), 7 November 1985 (07.11.85) A EP, Al, 490818 (SANDOZ-PATENT-GMBH), 17 June 1992 1-8 (17.06.92) A Chemical Abstracts, Volume 117, No 21, 1-8 23 November 1992 (23.11.92), (Columbus, Ohio, USA), Rudolphi, Karl A et al, "Adenosine a pharmacological concept for the treatment of cerebral ischemia", THE ABSTRACT No 204289v, Pharmacol. Cereb. Ischemia 1990, 439-448 Form PCTr;SAi210 (conunuauon of second sheet) (July 1992) INTERNA'1JCNA.L SPARC14 REPORT International 2pplication No. .PCT/D< 93/00158 Box I Observiations where cer-tan claims were found urisarchablc (Continuation of Item 1 or first sheet) This international sea rch report has not been establ ished in respect of certa in claims under Article 17(2)(a) for the folldwingreasons: 1. rWvi Claims Ncs.: 9-14 L.Jbecause they relate to subject mat-ter not required to be searched by this Authority, namely: See PCT Rule 39.l(I\J): Methods For t:reatment of the human or animal body by Surgery or therapy, as well as diagnostic methods. 2. ED claims Nos-- because they relate to parts of the international application that do not comply with the prescribed requirements to such an extent that21 no meaningful international search can be carried out, specifically: 3. Dl Claims Nos.: because they are dependent claims and are not drafted in accordance withi the second and third sentences of Rule 6.4(a). Dox 11 Observations where unity of Invention Is lacking (Continuation or item 2 or flrst shet) This International Searching Autboriy found multiple inventions in this international application, as follows: 1. F As all required additional search fees were timely paid by the applicant, this international search report covers all searchable claimos. 2. F1As all searcable cilms could be seariched without effori justifying an additional fee, this Authority did not invite payment of any additional fee. .FlAs only some of the required additional search fees were timely paid by the applicant, this international search report 3Elcovers only those claims for which fees were paid, specifically claims Nos., 4. No required additional search1 fees were timely paid by the applica2nt. Consequently, this international search report is Remark on Pi-lett I]lTe additional search fees were accompanied by the applicant's protest. I No protest accompanied the payment of additional search fees. Form PCTIISAJ210 (continuttion of first shet (July 1992) I NTERNAT1IONAL SEARCHl REPORT' Internaional application No, Information on patent family members 307/31P1DK9015 Patent document Publication Patent family Publication Cited in search report d ate meber(s) 7date US-A- 4791103 13/12/88 AU-B- 579412 24/11/88 AU-A- 4877485 01/05/86 CA-A- 1262898 14/11/89 EP-A,B- 0179667 30/04/86 SE-T3- 0179667 US-A- 3551409 29/12/70 CH-A- 495364 31/08/70 CH-A- 538485 15/08/73 OE-A- 1670175 05/11/70 FR-A- 1558462 28/02/69 GB-A- 1143150 00/00/00 NL-A- 6717061 24/06/68 US-A- 3502649 24/03/70 CH-A- 487926 31/03/70 OE-iA,B,C 1670077 13/08/70 FRl- 1538356 00/00/00 GB-'A- 1123245 00/00/00 NP-C- 128629 00/00/00 NL-A- 6706368 08/11/67 EP-A2- 0322242 28/06/89 AU-A- 2740188 29/06/89 AU-A- 8589491 12/12/91 BE-A- 1002167 28/08/90 CH-A- 677495 31/05/91 DE-A- 3843609 06/07/89 FR-A- 2629715 13/10/89 FR-A- 2663936 03/01/92 GB-A,B- 2212498 26/07/89 JP-A- 1203400 16/08/89 LU-A- 87414 07/07/89 NL-A- 8803140 17/07/89 SE-A- 8804609 21/12/88 US-A- 5032583 16/07/91 EP-A2- 0253962 27/01/88 SE-T3- 0253962 AU-B- 610822 30/05/91 AU-A- 7194687 29/10/87 DE-A- 3784977 29/04/93 JP-A- 62252726 04/11/87 US-A- 5023244 11/06/91 EP-A2- 0152944 28/08/85 OE-A- 3406S33 29/08/85 JP-A- 60193998 02/10/85 US-A- 4704381 03/11/87 DE-A- 2052596 04/05/72 AU-B- 4S0603 11/07/74 AU-A- 3497171 03/05/73 BE-A- 774399 25/04/72 CA-A- 983395 10/02/76 FR-A,B- 2111862 09/06/72 GB-A- 1325970 08/08/73 US-A- 3851056 26/11/74 Form I'Cl'ISA1210 (Patent family annex) (July 1992) INTE'RNATIONAL SEARCH R IEPORT International application No. Information on patent family members 3/79 C/K9/05 Patent document Publication Patent family Publication cited in search report date member(s) date OE-B- 1670077 14/08/74 CH-A- 487926 31/03/70 FR-A- 1538356 00/00/00 GB-A- 1123245 00/00/00 NL-C- 128629 00/00/00 NL-A- 6706368 08/11/67 US-A- 3502649 24/03/70 WO-Al- 8504882 07/11/85 AU-B- 582359 23/03/89 AU-A- M~9585 15/11/85 EP-A,B- 0180614 14/05/86 SE-T3- 0180614 EP-Al- 490818 17/06/92 NONE IPormn PCUdSA1210 (patent family annek) (July 1992)