ADENOSINE DERIVATIVES HAVING A2 AGONIST ACTIVITY

Abstract: Adenosine derivatives having A2 agonist activity of formula (I), a process for the preparation thereof and pharmaceutical compositions containing them. The compounds of the invention are useful for the treatment of cardiovascular pathologies and nervous system diseases.

Description: ADENOSINE DERIVATIVES HAVING A g AGONIST ACTIVITY The present invention relates to adenosine deriv tives having A 2 agonist activity and the use thereof therapy. Adenosine is known to modulate a number of physi logical functions. At the cardiovascular system leve adenosine is a strong vasodilator and a cardiac depre sor. On central nervous system, adenosine induces sed tive, anxiolytic and antiepileptic effects. At t kidney level, it exerts a diphasic action, inducing v soconstriction at low concentrations and vasodilatati at high doses. Adenosine acts as a lipolysis inhibit on fat cells and as an antiaggregant on platele (Stone T.W., Purine receptors and their pharmacologic roles. In: Advances in drug research. Academic Pre Limited, 1989, 18_, 291-429; Progress Cardiovasc. Di 1989, 32, 73-97). A number of studies showed adenosine actions a mediated by two subtypes of receptors which are locat on the cell membrane: an high-affinity one, inhibiti the activity of the enzyme adenylate cyclase ( receptor), and a low-affinity one, stimulating t activity of the same enzyme (A 2 receptor). (J. Me Chem. 1982, 5, 197-207. Physiol. Rev. 1990, 7)(3 761-845. J. Med. Chem. 1992, 3_5, 407-422). Both rece tors are widely spread in the different systems of t organism. In some tissues, however, only one of sa receptors is mainly present. For example, A, recept is prevailing at the cardiac level, whereas the receptor is present mainly at the vascular level a on platelets. Therefore, it is clear that compounds capable of interacting selectively with the A ? receptor could have an interesting pharmacological pattern. In fact, the vasodilating activity, together with the antiaggrega- ting action, can lead to useful therapeutical applica¬ tions in the treatment of severe cardiovascular patho¬ logies , such as ischemic cardiopathy, hypertension and atherosclerosis. Moreover, due to the actions on central nervous system, the use of A~ selective medicaments can be en¬ visaged in the treatment of cerebrovascular ischemia, epilepsy and various emotional disorders, such as anxiety and psychosis. The prototypical compound having activity on the A- receptor is adenosine-5'-N-ethyluronamide or NECA (Mol. Pharmacol., 1986, 2_5, 331-336). On the other hand, NECA is also active on the A, receptor and there¬ fore it lacks selectivity for the adenosine receptors. Being the only available compound having A- affinity, NECA was used for pharmacological tests for the receptor binding. Only recently, the use of NECA as a prototypical A 2 agonist has been gradually quit since compounds were found having a certain A 2 receptor selectivity. Said compounds are NECA derivatives which are substituted at the C2-position with phenylamino groups. For example, compound 2-(p-(carboxye- thyDphenylethylamino)-5'-N-ethyluronamide, named CGS 21680 (J. Pharmacol Exp. Ther., 1989, 251, 888-893) has become the reference compound for the pharmacological studies on A- receptor. Purine derivatives having a selective 2 agoni activity are disclosed, for example, in GB-A-2203149 EP-A-0309112, EP-A-0267878, EP-A-0277917, EP-A-0323807. Particularly, substitution at the 2-position the purine group has been considered promising to gi the desired selectivity (J. Med. Chem. 1992, 3_5, 407 422). 2-Alkynylpurine derivatives have been disclosed i EP-A-0219876. Now it has been found that 2-alkynyladenosine de rivatives substituted at the ethyne position with ary heterocyclic or hydroxyalkyl groups and in which t riboside residue is substituted by the N-alkyl- (or c cloalkyl)-uronamido, have surprisingly advantageo properties compared with the known compounds. The compounds of the invention have the followi general formula: NHR wherein R is hydrogen, C- ^ -Cg alkyl, C 3 -C 6 cycloalkyl, phen C 1 ~C 3 alk * l J R, has one of the following meanings: a) phenyl or naphthyl optionally substituted with one to three halogen atoms (chlorine, fluorine an . bromine), ι- c g alkyl, C ι -C 6 haloalkyl, c χ _c g alkoxy, C J ~ C Q haloalkoxy, C 2~ C 6 alkoxycarbonyl, C 2" C 6 al^oxyaikyl, ^-Cg alkylthio, thio, CHO, cyanomethyl, nitro, cyano, hydroxy, carboxy, C 2 ~ 6 acyl, amino, C.-C 3 monoalkylamino, C 2 -Cg dialkyla¬ mino, methylenedioxy; aminocarbonyl; b) a group of formula -(CH 2.) m-Het wherein m is 0 or an integer rom 1 to 3 and Het is 5 or 6 membere heterocyclic aromatic or non aromatic ring, optionally benzoσondensed, containing 1 to 3 heteroatoms selected from oxygen, nitrogen or sulphur, linked through a carbon atom or through nitrogen atom; c) C 3 ~ C 7 cycloalkyl optionally containing insatura tions or 2 -C . alkenyl; _ , ~5 d) -(CH, Δ) n -C-R i. R 4 R 2 is hydrogen, methyl or phenyl; R g is hydrogen, C,-C 8 linear or branched alkyl, C ς - cycloalkyl or C--C-, cycloalkenyl, phenyl-C,-C 2 -alkyl o R- and R 5 , taken together, form a 5 or 6-membere carbocyclic ring or R, is hydrogen and 2 and R. , take together, form an oxo group or a corresponding acetali derivative; when R is different from hydrogen and/or R^ i different from ethyl, R, can also be C,-C 8 linear o branched alkyl ; R , is OH , NH 2 , dialkylamino , halogen , cyano ; n is 0 or 1 to 4 ; Within the scope of the definitions given fo formula I, the following meanings are preferred: for C,-Cg alkyl: methyl or ethyl; for C--C 7 cycloalkyl: cyclopropyl, cyclopentyl, cyclohexyl or cyclohexenyl; for phenyl-C,-C 2 alkyl: benzyl or phenylethyl; for C.-Cg haloalkyl: trifluoromethyl; for halogen: chlorine; for C,-Cg alkoxy: methoxy or ethoxy; for C,-Cg haloalkoxy: trifluoromethoxy or difluo romethoxy; - for C ~ c 6 alkoxycarbonyl: methoxycarbonyl o ethoxycarbonyl; for C 2 -Cg alkoxyalkyl: methoxymethyl, methoxyethy or ethoxymeth l; for C 1 -Cg alkylthio: methylthio; - for C 2 -Cg acyl: acetyl; for C,-C 3 monoalkylamino: methylamino, ethyla ino isopropylamino; for C 2 -C 8 dialkylamino: dimethylamino, diethyla ino, methylethylamino, methylisopropylamino, dii sopropylamino; for heterocyclic aromatic or non aromatic rin . containing 1 to 3 N, S, 0 atoms: pyridyl, thienyl furyl, imidazolyl, thiazolyl, pyrazolyl, tria zolyl. - for acetalic derivative: diethylacetal. Preferred compounds of formula I are: 1. Compounds in which R is hydrogen; 2. Compounds in which R is C,-C 7 cycloalkyl; (R) or (S)-phenylisopropyl; 3. Compounds in which R, is phenyl or naphthyl optio- nally monosubstituted with one of the substituents in point a) ; 4. Compounds in which R, is phenyl optionally substi¬ tuted with one of the substituents in point a); 5. Compounds in which R, is phenyl, 2-, 3- or 4-ni- trophenyl, 2-, 3- or 4-aminophenyl, 2-, 3- or 4-methyl- phenyl, 2-, 3- or 4-acetylphenyl, 4-cyanomethylphenyl, 4-formylphenyl, 2-, 3- or 4-trifluoromethylphenyl, 2-, 3- or 4-fluorophenyl, 2-, 3- or 4-methoxyphenyl, 2-, 3- or 4-carbomethoxyphenyl, 2-, 3- or 4-carbethoxyphenyl; 6. Compounds in which R-, is cyclohexyl or 1-cy- clohexenyl; 7. Compounds in which R, is 2-thienyl, 2-pyridyl, 4- pyridyl, 4-pyrazolyl, 2-furyl, 3-furyl, 2-thiazolyl, 1- imidazolyl-methyl, 1-triazolylmethyl. R 5 I 8. Compounds in which R, is - (CH 2 ) -C-R 2 group I R 4 wherein R 2 is hydrogen or methyl, R 5 is hydrogen or C,-Cg alkyl, R. is hydroxy or amino. 9. Compounds in which R is hydrogen and R, has the meanings according to points 2-7 above. The claimed compounds can be used as anhydrous bases, solvates or pharmaceutically acceptable aci salts. Compounds I can be prepared according to th following general schemes: Scheme I hHR' Scheme II MR' In schemes I and II, R and R', have the sa meanings as R and R, or they are groups which can converted into R and R,, respectively, for example removing any protecting groups which can be present R* and R 1 , compatible with the reaction conditions; is Br or I and X is chlorine, bromine or iodine. Preferably Y is iodine and X is bromine or iodine. The reactions reported in schemes I and II ar carried out in the presence of catalysts (for example bis(triphenylphosphine) palladium dichloride and cuprous halide) and of a suitable acid-binding agent such as an organic base (for example: triethylamine diisopropylethylamine or pyridine) . As the solvent, a substituted amide (such as di methylformamide) , an ether (such as dioxane or tetrahy drofuran) , acetonitrile or optionally a mixture of tw or more of said solvents, are preferably used. The compounds of formula II, in which Y is iodin and R' is hydrogen, can be prepared from 2-iodioadeno sine (Synthesis, 1982, 670-672) according to th following scheme III e 2 The compounds of formula II, in which Y is iodine and R 1 is different from hydrogen can be prepared according to the following Scheme IV Scheme IV VI H.-N -R ( β ) - (Ph 3 P)Pd I 2 ; Cul ; E 3 N In the above scheme IV, R, R, and R 3 are as above defined. The compounds of formula VI are novel and they are a further object of the invention, as intermediates. The compounds of formula V are prepared by reaction of compounds II with an acetylene derivative, for example 1-trimethylsilylethynyle, under the condi¬ tions reported for the reaction between compounds II and III. Compounds V are novel and they are a further object of the invention, as intermediates. Compounds III and IV are known or they can be prepared according to well-known methods. Compounds I have a strong A 2 agonist selectivity and therefore they are useful for the treatment of car- diovascular pathologies such as cardiac ischemia, hy¬ pertension and atherosclerosis and of diseases of cen¬ tral nervous system such as cerebrovascular ischemia, epilepsy and emotional disorders (anxiety and psychosis) . Pharmacological activity The pharmacological properties of the disclosed compounds can be shown in the most suitable experimen¬ tal models both in vitro and in_ vivo. Adenosine A 2 receptor affinity was tested by means of receptor binding techniques on rat (Wistar strain) brain striatum, which is a tissue rich in A 2 receptors. Compound 3 H-CGS 21680 (J. Pharm. Exp. Ther. 1989, 251, 888-893) was used as the radioligand. The A, receptor affinity was tested with receptor binding techniques on rat (Wistar strain) cerebellar cortex membranes, which are tissues rich in A, recep- tors. 3H-Cyclohexyl-adenosine, 3H-CHA (Proc. Natl Acad. Sci. - USA - 1980, 77_, 5547-5551) was used as th radioligand. The affinity for the A, or A 2 recepto shown by each compound was compared to evaluate selec tivity for the A 2 receptor. A number of experimenta proofs evidence a marked relationship between th affinity found with binding techniques in brain tissue and the physiological effects modulated by adenosin receptors. In order to evaluate the functional response o specific tissues to the disclosed compounds, th isolated organs models were used. Particularly, the va sodilatation response, which is modulated by A receptors, was studied to the tested compounds (Eur. J Pharmacol 1990, 176, 207-21*2) on rat aorta in which contraction had been induced by a prostaglandi compound PGF 2 d . Any chronotropic negative effects o the various compounds (Br. J. Pharmacol .1983, 7_8_, 207 212) were tested on rat isolated heart atria, whos rate is known to be modulated by A, receptors. From th comparison of the activities shown by each compound fo the functional responses of the A 2 type (vasodilata tion) or the A. type (reduction in the heart rate) , th potential anti-ischemic and antihypertensive activitie of the tested compounds can be evaluated, as well a the absence of undesired effects on the heart rate. In order to evaluate the potential anti-athero sclerosis and anti-ischemic activities, the inhibitin effect of various compounds on platelet aggregatio induced by aggregation agents such as adenosine dipho sphate (ADP) (J. Physiol. 1963, 168, 178-195) wa studied. The test is particularly important as only th A 2 receptor is present on the platelet cell membrane. The ii vivo activity was evaluated in Swiss mic and spontaneously hypertensive rats (SHR). Th behaviour response to a treatment with various doses o the tested compounds administered parenterally was eva luated. To test the antiepileptic action, the propert of the novel compounds to antagonize the convulsion induced by pentylenetetrazole was evaluated in th mouse. The antihypertensive activity was tested measurin the systolic arterial pressure by means of the "tail cuff" technique (Arch. Int. Pharmacodyn., 1987, 286, 546-254) in SHR rats conditioned to the experimenta environment. The tested compounds were administered parente rally at increasing doses and arterial pressure an heart rate were measured at various times from the tre atment. The compounds of formula I showed a marked A affinity with Ki ranging from 7 to 200 nM. The A 2 se lectivity for some compounds is higher than 100 an anyhow it turned ut to be markedly higher than that o the prototypical compound NECA. In the platelet aggre gation test, said compounds proved to be effectiv anti-platelet aggregation agents, with IC 5Q s of 0,1-1 μM. The vasodilatation activity is clear, as evidence by ED 5Q S of 0,1-10 μM, whereas the same concentration did not change the heart rate of isolated atria. In th in vivo models, the tested compounds showed a depres sing activity on central nervous system, they antagoni zed the convulsions induced by pentylenetetrazole an reduced the arterial pressure without changing signifi cantly the heart rate. The compounds turned out to b active at doses from 0.001 to 3 mg/k intraperitoneally. For the envisaged therapeutical uses, compounds will be formulated as suitable pharmaceutica compositions, which can be administered, for example by the oral or parenteral routes, using known techni ques and excipients, as described for example i Remington's Pharmaceutical Sciences Handbook, Mack Pub Co., NY, USA, XVII ed. The daily dosage will depend, o course, on many factors (severity of the pathology t treat, patient conditions, toxicology and pharmacokine tic of the selected compound) but generally it wil range from 0,01 to 10 mg/kg of body weight, preferabl from 0,1 to 1 mg/kg, optionally subdivided in more ad ministrations. Examples of pharmaceutical composition comprise capsules, tablets, solutions, syrups, vials controlled-release forms and the like. The following examples illustrate the invention. EXAMPLE 1 N-ethyl-1'-deoxy-1*-(6-amino-2-iodo-9H-purin-9-yl)-β-D- ribofuranuronamide. a) A solution of 2 g (5.08 mmoles) of 2-iodoadenosin in 100 ml of acetone was added with 9.6 g of p-tolue nesulfonic acid. The reaction mixture was stirred a room temperature for 1 h, then, after addition of 15 of NaHC0 3 , it was stirred again for 3 h. The solid wa removed and washed 2 times with ethyl acetate and th filtrate was concentrated to dryness. The residue wa flash chromatographed on a silica gel column elutin with CHCl 3 -MeOH (99:1) to give 1.62 g (74%) of 6-amino 2-iodo-9-(2' ,3'-O-isopropylidene-β-D-ribofuranosyl)-9H- purine as a solid: m.p. 185-187°C; 1 H NMR (Me 2 SO-d 6 ) -5 1.33 and 1.54 (s, 3H each, C(CH 3 ) 2 3.53 (m, 2H, CH 2 -5'), 4.19 ( , IH, H-4 ' ) , 5.07 (t, IH OH), 4.93 (m, IH, H-3'), 5.27 (m, IH, H-2'), 6.05 (d J=2.5 Hz, IH, H-l'), 7.76 (s, 2H, NH 2 ) , 8.28 (s, IH, H 8 ) . Anal. ( 13 H i6 IN 5°4 ) C . ' H ' N * b) A stirred solution of 1.6 g (3.7 mmoles) of th product obtained in a) in 200 ml of H ? 0 was added wit 0.60 g of KOH and, dropwise, with a solution of 1.70 (10.8 mmoles) of KMnO. in 50 ml of H 2 0. The mixture wa left aside in the dark at room temperature for 20 h The reaction mixture was cooled to 5-10 e C and then de colorized with a solution of 4 ml of 30% H 2 0 2 in 16 m of water, keeping the temperature under.10°C with a ice-salt bath. The mixture was filtered through Celit and the filtrate was concentrated under vacuum to abou 15 ml and then acidified to pH 4 con 2N HCl. The resul ting precipitate was filtered, and subsequently washe with water, acetone and ether to give 1.25 g (76%) o 1 ■ -deoxy-1'-(6-amino-2-iodo-9H-purin-9-yl)-2 ' ,3•-O-iso- propylidene-β-D-ribofuranuronic acid as a white solid: m.p. 187-190°C; IR max 1590, 1640 cm "1 (COOH); 1 H NMR (Me 2 SO-dg) 5 1.33 and 1.49 (s, 3H each, C(CH 3 ) 2 , 4.64 (s, IH, H-4 » ) 5.35 (d, J 3 , 2 ,=5.6 Hz, IH, H-3' ) , 5.41 (d, J 2 , 3 ,=5. Hz, IH, H-2'), 6.23 (s, IH, H-l'), 7.53 (s, IH, COOH) 7.67 (s, 2H, NH 2 ), 8.17 (s, IH, H-8). Anal (C 13 H 14 IN 5°5 ) C ' H ' N* c) A solution of 1.72 g (3.85 mmoles) of the produc obtained in b) in 80 ml of 50% formic acid was stirre at 80 β C for 1.5 h. The reaction mixture was evaporate under vacuum and the residue was dissolved in water and the solution was evaporated. This process wa repeated several times, until there was no more odor o formic acid in the residue. Recrystallization fro water yielded 1.33 g (85%) of l'-deoxy-l'-(6-amino-2 iodo-9H-purin-9-yl)-β-D-ribofuranuronic acid as a whit solid: m.p. 217-220 e C (dec.) E NMR (Me 2 SO-d 6 ) 54.28 (m, IH, H-3 « ), 4.41 (d, J=2. Hz, IH, H-4'), 4.81 (m, IH, H-2 » ), 5.95 (d, J=6.7 Hz IH, H-l'), 7.78 (s, 2H, H 2 ) , 8.38 (s, IH, H-8) , 12.9 (br s, IH, COOH). Anal. (C 10 H 10 IN 5 5 ) C, H, N. d) A cooled (5°C) and stirred solution of 1.29 g (3.1 mmoles) of the product obtained in c) . in 150 ml o absolute ethanol was added dropwise with 1.15 ml o ice-cooled S0C1-. The mixture was stirred at room tem perature .overnight and then brought to pH 8 wit saturated aqueous sodium bicarbonate. The mixture wa filtered and the filtrate was concentrated unde vacuum. The recrystallization of the residue fro water-ethanol (1:1) gave 900 mg (65%) of ethyl l'-de oxy-1'-( -amino-2-iodo-9H-purin-9-yl)-β-D-ribofuranu- ronate as a white solid: m.p. 221-223°C (dec.) IR 0 max 1728 cm "1 (COOEt); 1 H NMR (Me 2 S0-Dg) 1.2 (t, 3H, -CH 2 CH 3 ), 4.18 (q, 2H, -CH 2 CH 3 ), 4.34 (m, IH H-3'), 4.47 (s, IH, H-4' ) , 4.58 (m, IH, H-2' ) , 5.96 (d J=6.7 Hz, IH, H-l'), 7.74 (s, 2H, NH 2 ), 8.33 (s, IH, H 8 ) . Anal. (c i2 H i4 IN 5 0 5* ) > H ' N - e) A mixture of 620 mg of the product obtained in th step d) and 18 ml of dry ethylamine was stirred a -20 β C for 3 h and then at room temperature overnight. The reaction mixture was diluted with absolute ethano and the precipitated product was filtered and washe with dry ether to give 530 mg (85%) of N-ethyl-1' deoxy-1'-(6-amino-2-iodo-9H-purin-9-yl)-β-D-ribofura- nurona ide as a pure solid: m.p. 232-234°C IR -0 max 1637, 1560 cm "1 (C=0, amide); 1 H NMR (Me 2 S0 dg) δ 1.06 (t, 3H, -CH 2 CH 3 ), 3.28 ( , 2H, -CH 2 CH 3 ), 4.16 (m, IH, H-3'), 4.31 (d, J=2,l Hz, IH, H-4' ), 4.5 (m, IH, H-2'), 5.91 (d, IH, J=7.3 Hz, H-l'), 7.79 (s, 2H, NH 2 ), 8.15 (t, IH, NH) , 8.40 (s, IH, H-8). Anal. (C 12 H 15 IN 6 0 4 ) C, H, N. EXAMPLE 2 A solution of 250 mg (0.58 mmole) of the product o Example 1 in 10 ml of dry acetonitrile, 5 ml of DMF an 2.5 ml of triethylamine under nitrogen was added wit 8.1 mg (0.0115 mmole) of bis(triphenylphosphi ne)palladium dichloride and 0.58 mg (0.003 mmole) o cuprous iodide. The mixture was added with 2.9 mmole of phenylacetylene and the reaction mixture was stirre under nitrogen atmosphere at room temperature for 2 h The solvent was evaporated off under vacuum and th residue was chromatographed on a silica gel column eluting with a chloroform-benzene-methanol 78:10:1 mixture, to give N-ethyl-l'-deoxy-l , -(6-amino-2 phenylethynyl-9H-purin-9-yl)-β-D-ribofuranuronamide (yield: 60%) : m.p. 175-178°C (dec) 1 H NMR (Me 2 SO-dg) S 1.02 (t, 3H, -CH 2 CH 3 ) , 3.28 ( , 2H, -CH 2 CH 3 ), 4.15 (m, IH, H-3 • ) , 4.31 (d, J=1.2 Hz, IH, H- 4 « ), 4.62 (m, IH, H-2 ' ) , 5.97 (d, IH, J=7.5 Hz, H-l'), 7.58 (d, J=7.8 Hz, 2H, H-Ph), 7.46 (m, 2H, H-Ph), 7.66 (s, 2H, NH 2 ), 8.49 (s, IH, H-8) , 8.60 (t, IH, NH) . Anal. (C 20 H 20 NgO 4 " H j O) C, H, N. EXAMPLES 3-35 By the same method of Example 2, the following compounds I, wherein R 3 is ethyl and R is hydrogen, were prepared. Ex. R, - m.p. ("O Yield Reaction ' (%) time (h) * 11 -C-Ph 203-205 (dec) 75 20 OH 12 HO(CH 2 ) 4 245-247 . 69 24 Ex. R. m.p. ( β C) Yield Reaction (%) time (h) 13 —ζ ~ 200-203 (dec) 68 6 14 3 16 20 48 3 20 245-248 (dec) 70 20 211-214 (dec) 52 20 OH 22 (CH 2 ) 3 OH 20 23 CH 2 -CH-CH 3 5 I OH 24 -CH-CH 2 -CH 3 I OH CH 3 CH 3 25 -CH-CH 2 -CH-CH 3 135-137 41 OH 26 OH 153-155 30 20 27 NH„ 211-213 76 30 >o Ex. R. m.p. ( β C) Yield Reaction (%) time (h) 28 CH 2 NH 2 52 36 61 20 65 20 56 48 53 53 16 53 16 Anal. (CHN) and H-NMR in agreement with the give formulas. EXAMPLE 36 a) N-ethyl-1'-deoxy-1'-[6-amino-2-ethynyl-9H-purin-9- 1.6 mg (3.96 mmoles) of the product obtained i Example 15 in 30 ml of methanol were added wit 0.32 g of KOH in 10 ml of methanol. The reactio mixture was stirred at room temperature for 1 and then the solvent was removed under vacuum. T residue was partitioned between ethyl acetate a water and the combined organic layers were dri (Na 2 S0 4 ) and evaporated to give 1.0 g (81%) of t title compound as a chro atographically pu solid: m.p.: 245-248 β C (dec) 1 H NMR (Me 2 SO-dg) 61.04 (t, 3H, -CH 2 CH 3 ) , 3. (m, 2H, -CH 2 CH 3 ), 4.10 (s, IH, -C=CH) , 4.12 ( IH, H-3'), 4.29 (d, J=1.5, IH, H-4• ) , 4.55 ( , I H-2'), 5.92 (d, IH, J=7.5 Hz, H-l'), 7.62 (s, 2 NH 2 ), 8.46 (s, IH, H-8), 8.63 (t, IH, NH) . Ana (C 14 H 16 N 6°4 * H 2 0) C ' H ' N ' b) N-ethyl-1'-deoxy-1'-{6-amino-2-(4-trifluorome- thy1)phenyl]ethynyl-9h-purin-9-yl}-β-D-ribofura- nuronamide. A solution of 200 mg (0.66 mmole) of the compou of step a) in 10 ml of dry acetonitrile, 5 ml DMF and 2.6 ml of triethylamine under nitrog atmosphere, was added with 9.24 mg (0.013 mmol of bis(triphenylphosphine)palladium dichloride a 0.66 mg (0.0035 mmole) of cuprous iodide. T mixture was added with 3.3 mmoles of trifluoromethyl-iodobenzene and the reacti mixture was stirred under nitrogen atmosphere f 1 h at 50°C. The solvent was evaporated und vacuum and the residue was chromatographed on silica gel column, eluting with chloroform-be zene-methanol (80:10:10). The title product w obtained in a 65% yield, m.p.: 224-227°C (dec) X H NMR (Me 2 SO-dg) <5 " 1.01 (t, 3H, -CH 2 CH 3 ) , 3.2 (m, 2H, -CH 2 CH 3 ), 4.14 (m, IH, H-3' ) , 4.31 (s, IH H-4 » ), 4.61 (m, IH, H-2'), 5.97 (d, IH, J=7.5 Hz H-l'), 7.71 (s, 2H, NH 2 ), 7.81 (s, 4H, H-Ph), 8.5 (s, IH, H-8), 8.58 (t, IH, NH) . Anal (C 21 H 19 3 Ng0 4 * H 2 0) C, H, N. EXAMPLES 37-43 By the same method of Example 36, the followin compounds I, wherein R is hydrogen and R, is ethyl were prepared. EXAMPLES 44-46 By reacting the compound of Example Id with a amine selected from: - cyclopentylamine; - benzylamine; - aniline; at room temperature for 20 hours, the followin compounds as chroma tographically pure oils wer obtained after the usual work-up (solution concentrate to dryness and the residue chroma tographed on a silic gel column eluting with a suitable mixture o solvents): N-cyclopentyl-1 ' -deoxy-1 ' - ( 6-amino-2-iodo-9H-purin-9- yl) -β-D-ribof uranuronamide (Ila) X H NMR (Me 2 SO-d 6 ) δ 1.33-1.98 (m, 8H, H-cyclopentyl) 4.06 ( , IH, H-cyclopentyl), 4.19 (m, IH, H-3'), 4.3 (d, IH, J = 2.4 Hz, H-4 » ), 4.58 (m, IH, H-2 ' ) , 5.93 (d IH, J = 6.4 Hz, H-l'), 7.77 (s, 2H, NH 2 ), 8.08 (d, IH J = 7.5 Hz, NH), 8.50 (s, IH, H-8) . Anal. ( c ι 5 H 19 IHg0 4 C, H, N. N-benzyl-1 ' -deoxy-1 • - ( 6-amino-2-iodo-9H-purin-9-yl ) -β- D-ribof uranuronamide ( lib ) X H NMR (Me 2 S0-dg) 64.22 (m, IH, H-3 ' ) , 4.42 (d, IH, = 1.5 Hz, H-4-), 4.46 (m, 2H, CH 2 ) , 4.59 (m, IH, H-2 ' ) 5.94 (d, IH, J = 7.2 Hz, H-l'), 7.28 (m, 5H, H-Ph) 7.81 (s, 2H, NH 2 ), 8.40 (s, IH, H-8) , 8.74 (t, IH, HN) Anal. ( c i7 H i7 IN 6°4^ > H > N ' N-phenyl-1 ' -deoxy-1 • - ( 6-amino-2-iodo-9H-purin-9-yl ) -β- D-ribof ranuronamide (lie) ■ '"H NMR (Me 2 S0-dg) <54.35 (m, IH, H-3' ) , 4.58 (d, J 1.5 Hz, IH, H-4 « ), 4.66 ( , IH, H-2'), 6.01 (d, IH, J 6.6 Hz, H-l'), 7.11 (t, IH, H-Ph), 7.34 (t, 2H, H-Ph), 7.65 (d, 2H, H-Ph), 7.78 (s, 2H, NH 2 ) , 8.52 (s, IH, H- 8), 10.16 (s, IH, NH) . Anal. ( c ι-s H ι 5 IN g0 4 ) c > H ' N - Said compounds were reacted with 1-hexyne according to the procedure of Example 2 to give the compounds of formula I reported in the following Table. Ex. R R, R 3 m.p. Yield Reaction ( β C) (%) time (h) 44 H -(CH 2 ) 3 -CH 3 _ 145-147 48 40 45 H -(CH 2 ) 3 -CH 3 -CH 2 - Q 128-130 37 40 46 H -(CH 2 ) 3 -CH 3 -(QS 35 40 Anal. (CHN) and H-NMR in agreement with the given formulas. EXAMPLE 47 a) N-ethyl-1'-deoxy-1'-(2,6-diiodo-9H-purin-9-yl)-β- D-ribofuranuronamide (VI) To 0.5 g (1.16 mmol) in 10 ml of DMF was added 1.7 ml of isopentyl nitrite and 5.6 ml of diiodometan and the mixture was heated at 85 e C under a N 2 atmosphere for 2 h. The solvent was removed unde pressure (oil pomp) and the residue wa chromatographed on a flash silica gel colum eluting with a gradient from CHC1 3 to CHCl 3 -MeO (98:2) to give 285 mg (45%) of the title compoun as a chromatographiσally pure oil. ! H NMR (Me 2 SO-dg) δ 1.04 (t, 3H, CH 2 CH 3 ) , 3.19 (m 2H, CH 2 CH 3 ), 4.28 (m, IH, H-3' ) , 4.36 (s, IH, H 4'), 4.70 (m, IH, H-2' ) , 6.03 (d, IH, J = 6.4 Hz H-l'), 8.15 (t, IH, NH), 8.98 (s, IH, H-8). Anal (C 12 H 13 I 2 N 5°4 ) C ' H ' N* b) N-ethyl-1'-deoxy-1'-(6-cyclopentylamino-2-iodo-9H- purin-9-yl)-β-D-ribofuranuronamide (VII) To 240 g (0.44 mmol) of N-ethyl-l'-deoxy-l'-(2,6 diiodo-9H-purin-9-yl)-β-D-ribofuranuronamide wa added 10 ml of eyelopentylamine and the mixtur was stirred at room temperature for 4 h. Th solution was concentrated to dryness and th residue was chromatographed on a silica gel colum eluting with CHCl 3 ~MeOH (96:4) to give 160 m (72%) of the title compound as chro atographically pure oil. ^ ■ H NMR (Me 2 SO-dg) δ 1.07 (t, 3H, CH 2 CH 3 ) , 1.48 2.08 (m, 8H, H-cyclopentyl), 3.23 (m, 2H, CH 2 CH 3 ) 4.18 (m, IH, H-3'), 4.32 (d, IH, J = 1.7 Hz, H 4 « ), 4.44 ( , IH, H-cyclopentyl), 4.60 (m, IH, H 2'), 5.92 (d, IH, J = 7.2 Hz, H-l'), 8.16 (t, IH NHCH 2 ), 8,29 (d, IH, J = 7.7 Hz, NH) , 8.41 (s, IH H-8). Anal. (C 17 H 23 INg0 4 ) C, H, N. c) N-ethyl-1•-deoxy-1'-[6-cyclopentylamino-2-(1- hexyn-l-yl)-9H-purin-9-yl]-β-D-ribofuranuronamide (VIII) The title compound was prepared according t Example 2. Reaction time: 16 h. Chromatographical system: chloroform-methano (95:5) Yield: 57% m.p. : 120-122 °C. ^ MR (Me 2 SO-dg) δ 0.92 (t, 3H, CH j CH- j ), 1.08 (t, 3H, NCH 2 CH 3 ), 1.35-2.07 (m, 12H, H-cyclopentyl and CH 2 CH 2 ), 2.44 (m, 2H, CH 2 C = C) , 3.31 (m, 2H, NCH 2 CH 3 ), 4.12 (m, IH, H-3 r ) , 4.31 (s, IH, H-4' ) , 4.60 (m, 2H, H-2 ' and H-cyclopentyl) , 5.94 (d, IH, J - 7.7 Hz, H-l'), 8.03 (d, IH, NH) , 8.43 (s, IH, H-8), 8.76 (t, IH, NHCH 2 ). Anal. ( C 2 3 H 32 N 6°4 " 10 H 2 0) C, H, N.

Claims: CLAIMS 1. Compounds of formula I NBR wherein R is hydrogen, C,-C 8 alkyl,* 3 -C 7 cycloalkyl, pheny R, has one of the following meanings: a) phenyl or naphthyl optionally substituted with on to three halogen atoms (chlorine, fluorine an bromine) , C,-Cg alkyl, C 1 "C 6 haloalkyl, C 1 "C alkoxy, C.-Cg haloalkoxy, C 2 -Cg alkoxycarbonyl C 2 -Cg alkoxyalkyl, C^Cg alkylthio, thio, CHO cyanomethyl, nitro, cyano, hydroxy, carboxy, C 2 -C acyl, amino, C,-C 3 monoalkylamino, C 2 -C 8 dialkyla mino, methylenedioxy; aminocarbonyl; b) a group of formula -(CH 2 ) m -Het wherein m is 0 o an integer from 1 to 3 and Het is 5 or 6 membere heterocyclic aromatic or non aromatic ring optionally benzocondensed, containing 1 to heteroatoms selected from oxygen, nitrogen o sulphur, linked through a carbon atom or through nitrogen atom; c) C 3 -C 7 cycloalkyl optionally containing insatura tions or 2 -C 4 alkenyl; R 5 I d) -(CH 2 ) n -C-R 2 I R 4 2 is hydrogen, methyl or phenyl; R e is hydrogen, C- j -Cg linear or branched alkyl, ^-C cycloalkyl or C 3 -C 6 cycloalkenyl, phenyl-C,-C 2 -alkyl o R 2 and R g , taken together, form a 5 or 6-membere carbocyclic ring or R- is hydrogen and R 2 and R 4 , take together, form an oxo group or a corresponding acetali derivative; when R is different from hydrogen and/or R 3 i different from ethyl, R, can also be .-Cg linear o branched alkyl; R 4 is OH, NH 2 , dialkylamino halogen, cyano; n is 0 or 1 to 4. 2. Compounds according to claim 1 in which R i hydrogen or cyclopentyl. 3. Compounds according to claim 1 or 2 in which R, i phenyl, 2-, 3- or 4-nitrophenyl, 2-, 3- or 4-ami nophenyl, 2-, 3- or 4-methylphenyl, 2-, 3- or 4-acetyl phenyl, 4-cyanometh lphenyl, 4-formylphenyl, 2-, 3- o 4-trifluoromethylphenyl, 2-, 3- or 4-fluorophenyl, 2- 3- or 4-methoxyphenyl, 2-, 3- or 4-carbomethoxyphenyl 2-, 3- or 4-carbethoxyphenyl, cyclohexyl or 1-cy clohexenyl, hydroxyphenyl. 4. Compounds according to claim 1 or 2 in which R, i 2-thienyl, 2-pyridyl, 4-pyridyl, 4-pyrazolyl, 2-furyl 3-furyl , 2-thiazolyl . 5. Compounds according to claim 1 or 2 in which R*, i a group R 5 I -(CH 2 ) -C-R 2 wherein R 2 and R 5 are hydrogen and n is an I OH integer 1 to 4 or R 2 is phenyl, Re is hydrogen and n is zero. 6. A compound according to anyone of the precedin claims, wherein R 3 is cyclopentyl, benzyl or phenyl. 7. A compound according to claim 6, wherein R. is n butyl. 8. A process for the preparation of compounds o formula I which comprises the reaction of a compound o formula II MR * wherein Y is Br or I and R ' has the same meanings as or is a group which can be converted into R, with compound of formula III R^-C≡≡C-H III wherein R 1 , has the same meanings as R, or is a grou which can be converted into R, . 9. A process for the preparation of compounds o formula I which comprises the reaction of a compound o formula IV R , 1 --X IV wherein R* is as defined in claim 8, with a compoun of formula V 20 wherein R' is as defined in the claim 8, 10. Compounds of formula V WR' wherein R' is as defined in claims 8 and 9, as interme diate compounds. 11. Compounds of formula VI I VI wherein R 3 is as defined in claim 1, as intermedia compound. 12. Pharmaceutical compositions containing a compou of claims 1-7 in admixture with a suitable carrier.

Inventors: CRISTALLI GLORIA, CRISTALLI, GLORIA
Applicants: CRISTALLI GLORIA, SCHERING PLOUGH S P A, SCHERING-PLOUGH S.P.A.
Application year: 1993
Main classification: C07H19
IPC codes: A61K31, A61P9, A61P25, C07H19