CA2220642A1 - C-4' modified adenosine kinase inhibitors - Google Patents

Abstract

This invention relates to adenosine kinase inhibitors and to nucleoside analogs, C-4' modified pyrrolo[2,3-d] pyrimidine and pyrazolo[3,4-d]
pyrimidine nucleoside analogs having activity as adenosine kinase inhibitors.
The invention relates to nucleoside analogs of this kind, having zero substitutions or two substitutions at the C-4' position of the furanose (sugar) moiety. The invention also relates to the preparation and use of these adenosine kinase inhibitors in the treatment of cardiovascular, and cerebrovascular diseases, inflammation and other diseases which can be regulated by increasing the local concentration of adenosine.

Description

W O 96/4070~ PCT~US96tlO404 C-4' MODIFIED ADENOSINE KINASE INHIBITORS

This invention relates to adenosine kinase inhibitors and to nucleoside analogs, C-4' modified pyrrolo[2,3-d] pyrimidine and pyrazolo[3,4-d]
pyrimidine nucleoside analogs having activity as adenosine kinase inhibitors.
5 The invention relates to nucleoside analogs of this kind, having zero substitutions or two substitutions at the C-4' position of the furanose (sugar) moiety. The invention also relates to the preparation and use of these adenosinekinase inhibitors in the treatment of cardiovascular, and cerebrovascular diseases, infla",r"dlion and other diseases which can be regulated by increasing10 the local concentration of adenosine.
This application is a continuation in part of Serial No.07/812,916, filed December 23, 1991, which is a continuation-in-part of Serial No.
07/647,117, filed January 23, 1991, which is a continuation-in-part of Serial No. 466,979, filed January 18, 1990; which is a continuation-in-part of Serial 15 No.408,707, filed September 15,1989. This appiication is also a continuation-in-part of Serial No. 08/191,282. The disclosures of these applications are incorporated herein by reference.

Sl~BSTlTUTE SHEET (RULE 26) W O ~G/4G70~ PCT~US96/10404 BACKGROUND OF THE INVENTIC)N
Adenosine is an endogenously produced molecule that plays a major role in a variety of important cellular processes It is a v~sor~ tor, can inhibit immune function, enhance activation of mast cells (associated with allergic 5 reactions), inhibit neutrophil oxygen free-radical production, is antiarrhythmic, and is an inhibitory neurotransmitter. Adenosine is phosphorylated to adenosine triphospl1aLe (ATP) which is used by all cells to store energy for use in future energy-utilizing metabolic reactions or mechanical work (e.g. muscle contraction).
Extracellular adel ,osine, frequently prouc ed by breakdown of intracellular ATP pools, 10 evokes a variety of pharmacological responses through activation of extracellular adenosine receptors located on the surface of nearly all cells. For example, adenosine produces a variety of cardiovascular related effects including vasodilation, inhibition of platelet aggregation, and negative inotropic, c~ ,ru, luLI .~ i and domoll opic effects on the heart. Adenosine also has effects within the central nervous system 15 (CNS) including inhibition of neulull~l-sll,ilLer release from presynaptic neurons and inhibition of post-synaptic neuron firing in brain and the spinal cord and at sites of inflammation, such as inhibition of neutrophil adhesion to endothelial cells andinhibition of neutrophil oxygen free-radical production.
Compounds that increase extracellular adenosine can be beneficial to 20 living organisms, particularly under certain conditions. For example, com~.ounds that increase adenosine levels have been associated with the treatment of ischemic conditions such as stroke, as well as other conditions benefitted by enhanced adenosine levels, such as inflammation, alll.liLis, seizures, epilepsy and otherneul .,lo~ l co, IdiLiCJllS. The CCJII Ipounds are also useful for treating pain, as muscle 25 relaxants, and for inducing sieep.
Adenosine kinase is a cytosolic enzyme which catalyzes the phospl ,olylation of adenosine to AMP. Inhibition of ade"osi"e kinase can potentially reduce the ability of the cell to utilize adenosine, leading to increased adenosine outc~ide of the cell where it is pharmacologically active. However, the regulation of 30 adenosine concentration is complex and involves other adenosine-metabolizing enzymes each with different kinetic properties and mechanisms of regulation.
Adenosine can also be deaminated to inosine by adenosine deaminase (ADA) and CA 02220642 l997-ll-28 W O 96/40705 PCT~US96/10404 condensed with L-homocysteine to S-adenosylhomocysteine (SAH) by SAH
hydrolase. The role of each of these enzymes in mo~lul~ting adenosine concentration is dependent on the prevailing physiological conditions, is tissuespecific and is not well understood.
A number of nucleosides including pyrrolo[2,3-d]pyrimidine and pyrazolo[3,4-d]pyrimidine analogs have been evaluated for inhibition of adenosine kinase but were reported to have IC,-'s of greater than 800 nM. Caldwell and Henderson, Cancer Cl ,emull ,er. Rep. 2:237-46 (1971); Miller et al., J. Biol. Chem.
254:2346-52 (1979). A few compounds have been reported as potent inhibitors of 10 adenosine kinase with ~'s of less than 100 nM. These are the purine nucleosides, 5'-amino-5'-deoxyadenosine (Miller et al.) and 1,12-bis(adenosin-N6-yl)dodecane (Prescott et al., Nucleosides & Nucleotides. 8:297 (1989)); and the pyrrolopyrimidine nucleosides, 5-iodotl ~hercidin (Henderson et al., Cancer Chemotherapy Rep. Part 2 3:71-85 (1972); Bot,Lei",ùs et al., Proc. Natl. Acad. Sci. USA. 80:2829-33 (1983);
15 Davies et al., Biochem. Pharmacol. 35:3021-29 (1986)) and 5'-deoxy-5-iodotuberc;din (Davies et al., Biochem. Phar" ,acol., 33:347-55 (1984) and 35:3021 -29 (1986))-Some of these compounds have been used to evaluate whetheradenosine kinase inhibition might lead to increased extracellular adenosine 20 concentrations. In rat cardiomyocytes, inhibition of adenosine deaminase by 2'-deox~corur"~cin was reported to have no effect on adenosine release from the cells.
In contrast, inhibition of ADA together with adenosine kinase by 5'-amino-5'-deoxyadenosi"e resulted in a 6-fold increase in adenosine release. Zoref-Shani et al., J. Mol Cell. Cardiol.. 20:23-33 (1988). The effects of the adenosine kinase25 i"l ,ibiL ,r alone were not, epo, led. Similar results were reported in isol~ed ~uinea pig hearts; in these studies addition of 5'-amino-5'-deoxyadenosine to the perfusionmedium, in the presence of EHNA to inhibit deamination, was reported to result in a 15-fold increase of adenosine release. Schrader in Regulatory Funct1On of Adenosine: (Beme et al.) eds. pp.133-156 (1983). These effects were not apparent30 in the absence of ADA inhibition, and other studies using isol~t~d rat hearts perfused with 5-iodotubercidin alone, have reported no increase in perfusate adenosine concentration under normoxic conditions Newby et al., Biochem. J.. 214:317-323 (1983), or under hypoxic, anoxic or ischemic condiLic, Is, Achtenberg et al., Biochem.
J., 235:13-17 (1986). In other studies, adenosine release has been measured in neuroblastoma cells in culture and co""~ared with that of a variant deficient inadenosine kinase (AK-). The AK cells used in this shldy were said to release 5 adenosine at an accelerated rate; the concentration of adenosine in the growthmedium was reported to be elevated compared to the normal cells. Green, J.
Supramol. Structure. 13:175-182 (1980). In rat and guinea pig brain slices, adenosine uptake was reportedly inhibited by the adenosine kinase inhibitors, 5-iodotubercidin and 5'-deoxy-5-iodotubercidin. Davis et al., Biochem. Pl,ar")acol..
10 33:347-55 (1984). However, inhibition of uptake and intracellular trapping via phosphorylation does not necess~rily result in increased extracellular adenosine, since the adenosine could enter other metabolic pathways or the percentage of adenosine being phosphorylated could be insignificant compared to the total adenosine removed.
The e~fects of ade"osi"e and certain inhibitor~; of ade"osi"e ~t~holj.sm, including 5-iodotubercidin were ev~ ted in an experimental model in which dog hearts were s~ b;~ te~l to ischemia and reperfusion; 5-iodotubercidin was reported to have inconsistent effects. Wu, et al., Cytobios. 50:7-12 (1987).
Although the adenosine kinase il IhiLJilol -~i, 5'-amino-5'-deoxyadenosine 20 and 5-iodotubercidin have been widely used in experimental models, the susce~ Libility of 5'-amino-5'-deoxyadenosine to dea" ,i"aLion, and hence its potentially short half life, and the cytotoxicity of 5-iodotuber~,;di" make ~heir clinical utility limited and may limit inter~r~taLions based on these compounds The known pyrrolo[2,3-d]pyrimidines,5-iodotul.er~;d;n and 5'-deoxy-5-iodotubercidin have been reported to 25 cause pronounced general flaccidity and much-reduced spontaneous locomotor activity in mice, interpreted to be skeletal muscle relaxation; to cause hypothermia in mice; and to decrease blood pressure and heart rate in anesthetized rats. Daves et al., Biochem. Pharmacol.. 33:347-55 (1984) and 35:3021-29 (1986); and U.S.
- Patent No. 4,455,420). The skeletal muscle effects of these compounds have been 30 poorly documented, while the other effects were considered significant toxicities.

W O ~C/~ ; PCT~US96/10404 More recent rer~ "ces co"ce" ,ed with the mechanisms and effects of adenosine kinase inhibitors are Keil et al., Life Sciences 51:171-76 (1992); Zhang et al., J.Pl,a""acol. Exper. Ther. 264(3): 1415 (1993); Phillis et al., Life Sciences. 53:
497-502 (1993); Sciotti etal., J. Cerebral Blood FlowMetab.,13:201-207 (1993); Pak 5 et al., Soc. for Neuroscience Abs20: 149.2 (1994); White, Soc. Neurosci. Abs.,20:308.9 (1994); and Firestein etal., J. Immunology 154:326-34 (1995). These publications in gel ,er~l show that ade"osi"e kinase i, lhibiLol :j, as a class, have a role in brain fu, IcLio"s, and show pron~ise in connection with the treatment of neurological conditions such as seizures. One ,~rer~nce, Phillis et al., indicates that the known adenosine kinase inhibitor 5-iodotubercidin apparently does not protect against ischemic cer~bral injury. Keil et al. disclose that adenosine kinase plays a key role in the mediation of nervous system responses to stimulus, particularly pain (antinociceplion), but notes that the control of endogenous adenosine concentrations by such means is a complex process requiring further study.
Thus, there is a need for selective, potent, and bioavailable adenosine kinase inhibitors with a useful half-life, i.e. compounds which can be exploited to benericially influence or control endogenous adenosine kinase activity, and Ll ,er~rore, extracellular adenosine levels. The compounds of the invention are suitable adenosine kinase inhibitors having these characteristics.

SUMMARY OF THE INVENTION
The invention is directed to novel pyrrolo[2,3-d] pyrimidine or pyrazolo[3,4-d] pyrimidine nucleoside analogs having activity as adenosine kinase i, lhibiLo, ~, wherein the furanose moiety has zero substituents or two substituents at the C-4' position. Preferred substitutents are hydroxymethyl, aminomethyl, and methyl. Most preferred are compounds where both substituents are the same, but are not both methyl, or both substituents form a small ring, such as cyclopropyl. In addition to the furanose moiety, additional asymmetric carbons may be present incompounds of the present invention, for example in the substituted heterocyclic pyrrolo[2,3-d] pyrimidine or pyrazolo[3,4-d]pyrimidine ring. All of the resulting isomers, en~"liomers, and diastereomers are considered to fall within the scope of the present invention.

These compounds are selective adenosine kinase inhibitors with pult:l ,cies comp~l able to or significantly higher than other known adenosine kinase inhibitors. The compounds are also nontoxic, particularly in connection with liver function. The invention concerns the compounds themselves, the preparation of 5 these compounds, and the in vitro and in vivo adenosine kinase inhibition activity of these compounds. Another aspect of the invention is dire!cted to the clinical use of the compounds to increase adenosine concentrations in biological systems. For example, in vivo inhibition of adenosine kinase prevents phosphorylation of adenosine resulting in higher local concel Ill ~Lions of endogenous adenosine.
The compounds of the invention possess advantages for pharmaceutical use such as enhanced pharmacological selectivity, efficacy, bioavailability, ease of manufacture and compound stability.
The compounds of the invention may be usecl clinically to treat medical conditions where an increased localized adenosine concer.LlaLion is beneficial.
1~; Accordil IYlr- the invention is directed to the Ll ~LI I ,e, IL of ischemic conditions such as stroke, as well as other conditions benefitted by e, Ihal ,ced adel losine levels, such as inflammation, arthritis, seizures, epilepsy and other neurological conditions. The compounds are also useful for treating pain, as muscle re!laxants, and for inducing sleep.
The invention is also directed to prodrugs and pharmaceutically accepLable salts of the compounds described, and to pharmaceutical compositions suitable for different routes of drug administration and which comprise a therapeutically effective amount of a described compound admixed with a pharmacologically acceptable carrier.

W O 96/40705 PCT~US96/10404 Definitions The following terms generally have the following meanings.
The term "aryl" refers to aro",alic groups, which have at least one ring having a conjugated pi electron system, including for example carbocyclic aryl, 5 heterocyclic aryl and biaryl groups, all of which may be optionally substituted.
Carbocyclic aryl groups are groups wherein all the ring atoms on the aromatic ring are carbon atoms, such as phenyl. Also included are optionally substituted phenyl groups, being preferably phenyl or phenyl substituted by one to three substituents, preferably lower alkyl, hydroxy, lower alkoxy, lower alkanoyloxy, halogen, cyano, 10 perhalo lower alkyl, lower acylamino, lower alkoxycarbonyl, amino, alkylamino, carboxdl"ido, and sulfamido. Further included are phenyl rings fused with a five or six membered I ,eleroc~clic aryl or carbocyclic ring, optionally containing one or more heteroatoms such as oxygen, sulfur, or nitrogen.
Heterocyclic aryl groups are groups having from 1 to 4 heteroatoms as 15 ring atoms in the aromatic ring and the remainder of the ring atoms carbon atoms.
Suitable heteroatoms include oxygen, sulfur, and nitrogen, and include furanyl, thienyl, pyridyl, pyrrolyl, pyrimidyl, pyrazinyl, imidazolyl, and the like, all optionally substituted.
Optionally substituted furanyl represents 2- or 3-furanyl or 2- or 3-20 furanyl ,u, ~re, ~bly s~ ~hstituted by lower alkyl or halogen. Optionally s~ Ihstih Ited pyridylrepresents 2-, 3- or 4-pyridyl or 2-, 3- or 4-pyridyl preferably substituted by lower alkyl or halogen. Optionally sl ~hstitl ~ted thienyl represenl~ 2- or 3-thienyl, or 2- or 3-thienyl preferably substituted by lower alkyl or halogen.
The term "biaryl" represents phenyl substituted by carbocyclic aryl or 25 heterocyclic aryl as defined herein, ortho, meta or para to the point of attachment of the phenyl ring, advantageously para; biaryl is also represented as the -C6H4-Arsubstituent where Ar is aryl.
The term "aralkyl" refers to an alkyl group substituted with an aryl group. Suitable aralkyl groups include benzyl, picolyl, and the like, and may be30 optionally substituted.

The term "lower" rerel ~ ed to herein in connection with organic r~ r~ls or compounds, ~specLi~/ely defines such with up to and including 7, prerer~bly up to and including 4 and adv~ Jeol ~cly one or two carbon atoms. Such groups may be straight chain or brancl ,ed.
~i The terms (a) "alkyl amino", (b) "arylamino~', and (c) "aralkylamino", ,t:spe~;Li~ely, referto the groups -NRR' wherein respectively, (a) R is alkyl and R' is hydrogen, aryl or alkyl; (b) R is aryl and R' is hydrogen or aryl, and (c) R is aralkyl and R' is hydroge, I or aralkyl.
The term "acylamino" refers to RC(O)NR'.
The term "carbonyl" refers to -C(0)-.
The term Uacyl" refers to RC(O)- where R is alkyl, aryl, aralkyl, or alkenyl.
The term "carboxamide" or "carboxamido" refers to -CONR2 wherein each R is independently hydrogen, lower alkyl or lower aryl.
1~ The term "alkyl" refers to saturated aliphatic groups including straight-chain, b,cnched chain and cyclic groups, optionally containing one or more heteroatoms.
The term "alkenyl" refers to unsaturated alkyl groups which conlai" at least one carbon~arbon double bond and includes sL, ~igl ,L-chain, L,l c- ,cl ,ed or cyclic groups, optionally containing one or more heteroaLon,s such as oxygen, sulfur, or nitrogen.
The term "alkynyl" refers to unsaturated alkyl groups which contain at least one carbon carbon triple bond and includes straight c:hain, branched, or cyclic groups, optionally containing one or more heteroatoms such as oxygen, sulfur, or2~ nitrogen.
The term Uamidino'' refers to -C(NH)NH2 The term ~amidoximo" refers to -C(NOH)NH2 The term Uguanidino'' refers to -NR,CN(R2)NR3R4 where R" R2, R3 and R4 are independently hydrogen, alkyl or aryl groups.
The term Uaminoguanidino" refers to the group -NR,NR2CN(R3)NR4R5 where R" R2, R3, R4 and R5 are independently hydrogenl alkyl or aryl groups.

W O 96/40705 PCT~US96/10404 The term "ureido" refers to the group -NR,C(O)NR2R3where R" R2 and R3 are independently hydrogen, alkyl or aryl groups.
~ The term Ucarboxylic acid" refers to the group -COOH.
The term Uacylguanidino'' refers to the group -C(O)NR,CN(R2)NR3R4 5 where R" R2, R3 and R4 are independently hydrogen, alkyl or aryl groups.
The term "mercapto" refers to SH or a tautomeric form thereof.
The temm "alkylene" refers to a divalent straight chain or branched chain saturated aliphatic radical.
The temm Usulrona, I ,ido" means -SO2NHR where R is h~dl uge., or lower 1 0 alkyl.
The temm "N-sulfonyi amine" means -NHS02R where R is fluoro, lower peRluoroalkyl or lower alkyl.
The term UN-acylated sulfonamide" refers to the group -S02NHCOR
where R is lower alkyl or lower perfluoroalkyl.
The term Ubasic nitrogen" generally refers to the nitrogen atom of an alkyl amine and implies a compound whose conjug~ d acid in aqueous solution has a pKa in the range of 9 to 11.
The term "prodrug" refers to any co" ,pound that may have less i, ILI irlSic activity than the "drug" but when administered to a biological system generates the 20 "drug" sub~l~"ce either as a result of spontaneous chemical reaction or by enzyme catalyzed or ~, lel~bolic reaction. Reference is made to various prodrugs such as acyl esters, carbonates, and urethanes, included herein as examples. The groups illustrated are exemplary, not exhaustive and one skilled in the art could pr~,uare other known varieties of prodrugs. Such prodrugs fall within the scope of the 25 invention.
The term "pharmaceutically acceptable salt" includes salts of compounds described herein derived from the combination of a compound of this invention and an organic or inorga"ic acid. The compounds of the present invention are useful in both free base and salt form. A water solubilizing group is a group that CA 02220642 1997-ll-28 increases the so! Ihility of an inhibitor by a factor of at least 10 and pr~r~r~bly at least 100 at pH values suitable for intravenous adm i~ L, aliGrl (pH 4 to pH 10). In practice the use of salt form amounts to use of base form; both forms are within the scope of the present invention.
The term ~ ~ll "ent includes prophylactic or therapeutic administration of compounds of the invention, for the cure or amelioration of disease or s~""pto~,s associated with disease, and includes any benefit obtained or derived from the administration of the described compounds.

=

W O 96/40705 PCT~US96/10404 DETAILED DESCRIPTION OF THE INVENTION
The invention relates to C4'-modified pyrrolo~2,3-d]pyrimidine and pyrazolo[3,4-d]pyrimidine nucleoside analogs of Formula 1, having activity as adenosine kinase inhibitors.
F

G~N' 'E

A~
Z1 (~ ~5Z2 wherein:
A and B are both hydrogen, or are each independently alkenyl, the group (CH2)nQ, where n is from 1 to 4 and Q is hydrogen, hydroxy, alkyl, alkoxy, amino, azido, or halogen; or A and B together form a ring of from 3 to 6 carbons, the ring containing 0 to 3 heteroatoms selected from oxygen and nitrogen and optionally substituted by Q as defined above;
D is halogen, aryl, aralkyl, alkyl, alkenyl, alkynyl optionally containing one or more heteroatoms such as nitrogen, oxygen or sulfur, haloalkyl, cyano, or carboxamido;
1~ E is nothing when Y is r ~ oge,1; and is hydrogen, halogen, or alkyl when Y is carbon;
F is alkyl, aryl, aralkyl, halogen, amino, alkylamino, arylamino, aralkylamino, alkoxy, aryloxy, aralkyloxy, alkylthio, arylthio, aralkylthio;
~ G is hydrogen or halogen;
Y is carbon or nitrogen;
Z, and Z2 are independently hydrogen, acyl, or taken together form a cyclic carbonate; and pharmaceutically acceptable salts thereof.

WO 96/40705 PCT~US96/10404 rerably, A and B are the same, but are not both methyl, and most prerer~bly are hydrogen or (CH2)nQ where n is 1 and Q is hydroxy, or amino. Alsoprefered are compounds where A and B form a ring of frorn 3 to 4 carbons having 0 or 1 heteruaLo, "s. Where A and B are not the same, they are each chosen from the 5 group consisting of methyl, CH2OH, Ci~OC~ and Ci-2i Ni-2i . Z is prererably hydrogen, or in prodrug form is preferably acyl or carbonate ester.
D is pr~rer~bly haloye, ., heterocyclic aryl, phenyl or sl Ihstitl Ited phenyl;
E is nothing when Y is nitrogen and is prerer~bly hydrogen when Y is carbon;
G is preferably hydrogen; and F is halogen, amino, arylamino, or heterocyclic arylamino, most preferably phenylamino or substituted phenylamino. P,erer,ed substitutions are halogen, alkyl, alkoxy, or alkylamino or other groups containing a basic or acidic functionality that improves water solubility. The most ~rt:r~:n ~d substitution is at the 15 para position of phenylamino. For example, prerered col,lpounds of the invention include those where F is phenylamino, substituted at the para position with halogen (e.g. fluorine) or a water-solubilizing group.
Exemplary substitutions of the arylamino or phenylamino (Group F) which improve water solubility have the formula (CH2)rT where r is from 0 to 3 and 20 T is an alkyl chain of 0 to 16 carbon atoms containing one or more nitrogen atoms, N-sulfonylated amino, admidoximo, N-aminoguanidino, amidino, guanidino, acyl guallidillo, cyclic derivative of amidino, guanidino, or aminoguanidino, a heterocyclic aryl group, or a ~ or 6 membered alicyclic ring containing at least one basic nitrogen and optionally one or more oxygen atoms, and optionally substituted by CONW', 2~ where each V is independently an alkyl chain, at least one of which contains one or more basic nitrogen atoms, and optionaliy oxygen atoms, or V and V' together form a six-membered ring containing at least one basic nitrogen and optionally one or- more oxygen atoms. Additionally the water solubilizing groups T can be an anionic group such as sulfonic acid, carboxylic acid, squaric acid derivatives, ~-lell a~Olyl and other bioisosteric repiace, I~e, lls of a carboxylic acid group such as, but not limited to, those described in Carini et al, (J. Med. Chem 34, 2525 (1991)) and rererences cited therein. Similar substitutions can also be made at Group D to improve water 5 solubility.
It wiil be u"de,-~lood that compounds according to the invention, when made according to the methods set forth below, or by other methods, may be provided in both diastereomeric forms. Usually, one form will predominate in thereaction mixture, however, both forms are within the scope of the invention.
Prodrugs of the compounds of the pl-esenl invention are incl~ ed in the scope of this application. Such prodrugs may be prepared by esterification of the hydroxyl groups on the sugar ring. Specially preferred will be the ester derivatives that improve the water solubility properties.

SYNTHESIS OF ADENOSINE KINASE INHIBITORS
1~ The compounds of the invention can be made by several reaction schemes. Exemplary synthetic routes are given below.
The synthesis of compounds of the present invention can be viewed as consisting of the following steps: (A) preparation of the carbohydrate 2, (B) preparation of the heterocycle 3, (C) coupling of the carbohydrate and the 20 heterocycle to provide a protected intermediate 4, (D) modiricalion of substituents on the heterocycle and carbohydrate; and (E) removal of the protecting groups (Scheme 1). Each step is discussed below.

B O F D , B~y/ E ~ ~Y(DE
~NH /~E A~i A~

SC~3EME 1 W096/40705 PCT~US96/10404 (A) PREPARATION OF THE CARBOHYDRATE.
4-s~ ~hstifl Ited carbohydrates of formula 2 are used for the synthesis of compounds of Formula 1, where A and B are both hyd, uge, I or cl ,osen i. Idepe, Ide- Itly from methyl, azidomethyl, aminomethyl, alkylaminomethyl, alkoxymethyl, 5 hydroxymethyl or alkylthiomethyl. The formula 2 carbohyclrates are made from the known methyl 2,3-O-methylethylidenefuranoside 5 (Scheme 2). See, Leonard N. J.
et al. J. Heferocycl. Chem. 3, 485 (1966). The 5-alkoxy group is introduced to 5, to make 6, by the IIl~lo~ of Snyder J. R. ef al. Carbohydr. Res. 163, 169 (1987). The 5-deoxy, azido, amino, alkylamino, alkylthio and alternatively alkoxy c.arbohydrates 10 are made by first transforming the 5-hydroxy into a leaving group L, preferably mesylate, tosylate, trifluoromethanesulfonate or halide. Treatment of 7 with a nucleophile, e.g. hydride, alkylamine, dialkylamine, alkymerca,. ta.., alcohol or other precursors of amines such as azides or protected amines provides intermerli~tes of formula 8. The isopropylidene is then replaced for less reactive ~rotectil)g groups, 15 ,~r~rtrdbly benzyl, according to methods well known to those skilled in the art. For example, Greene T. W., Protective Groups in Organic Chemistry, John Wiley & Sons, New York (1981).

HO o R~ \~O~_OCH3 \_J NaH, R-L \--X

5 \ 6 H, A~ ~ oCH~ A~ ~ OC~

7 ~ 9 Carbohydrates for compounds of Formula 1 where A is hydroxymethyl 20 are made by the method of Barker R. et a/. J. Org. Chem. 26, 4605 (1961), to give compounds of formula 9 where A' is preferably benzyl protected hydroxymethyl.

W O 9~'~10705 PCT~US96/10404 The carbohydrate of Formula 2 is ~referably prepared by the method illustrated in Scheme 3. Treatment of methyl glycoside 9 with a thiol or a dithiol, ~,e~er~bly 1,3-,uropa"ediLl"cl, in the presence of an acid or a Lewis acid, prerer~bly boron trifluoride-diethyletherate, gives dithioacetal 10. Oxidation of the genera~e 5 alcohol using well described methods and reagents, e.g. pyridinium dich,omale,pyridinium chlGr~,cl " u" ,cle, Moffat oxidation, sulfur Ll io~id~pyridine, p,er~, d~ y Swem oxidation, gives ketone 11. Chelation-controlled addition of an orga"o",etallic B'-M, preferably organolithium, stereoselectively provides tertiary alcohol 12. The dithio~cePI protecting group is removed using a modification of the procedure 10 developed by Fetizon M. ef al. J. Chem. Soc. Chem. Comm. 382 (1972), involving treatment of l hlQ::~C.e~ l with iodomethane and an inorganic base, preferably calcium ca, bo"ale. Altematively, other dithio~cet~l dep, u~ec;~iol " rocedures are known which use reagents such as N-halosu~ci"i" ,ide, cupric, mercuric and silver salts. However the use of these oxidative procedures is precluded for compounds bearing 15 incompatible functional groups such as thioethers, azides or amines.

~~ RSH, IA A~ Oxlda~don A~ sR
P~ OP2 or H~ ~Pl SR ~ ~P1 R

B'M A~SR D~ OH
~P1 R p~ OP2 Altematively the second 4-C-substituent may be introduced using the procedure of Youssefyeh R. D. et al. J. Org Chem. 44(8), 1301(1g79) where alkylation of an aldehyde of formula 1~ with electrophile B'+ followed by a reduction 20 gives co"lpound 16 (Scheme 4). The aldehyde is obtained from the oxidative cleavage, preferably with sodium periodate, of hexofuranose 13 or oxidation of the CA 02220642 l997-ll-28 primary alcohol of furanoside 14, plererably using a hloffat oxidation. Another method to obtain compounds of formula (16) is to use the procedure of Johnson C
R. et al. J. Org Chem. 59(20), 5854 (1994).
HO~
HO~ oR
NalO4 o \~ H~ Base, B'~- HOX~_OR
P10 OP2 Reduction P~ ~P2 HO~ OR ~ 15 16 Oxidation p~ ~P2 ~i SCHEME 4 Carbohydrates for compounds of Formula 'I where A=B=H are made using a suitably protected carbohydrate of formula 2 where A'=B'=H. This carbohydrate is easily obtained from erythrofuranose accorcJil,g to methods wellknown to those skilled in the art described in Greene T. W., Profecfive Groups in 10 Organic Chemisfry, John Wiley & Sons, New York (1981). This carbohydrate is also easily obtained by reduction of the corresponding lactone as in N. Cohen et a/. J. Am.
Chem.Soc. 105, 3661, (1983).
Carbohydrates of formula 2, where A and B form a ring, e.g.
cyclopropyl, are prepared from D-ribose via the well known enol ether 17 (Scheme15 5). Inokawa S. ef al. Carbohydr. Res. 30, 127 (1973). Cyclopropanation is pe,r.""ed according to the procedure of Simmons H.E. et al. J. Am Chem. Soc. 81,4256 (1959) or one of its many modifications. Alterna~ively cyclopro~a"~lion is accomplished with a ~ o~lkane and a metal salt, preferably palladium. Cossy J.
ef al. Tefrahedron Leff. 28(39), 4547 (1987).

W O 96/40705 PCT~US96/10404 H~o OH O OC~ O ~ OH
~~ Cy:do~.upan~ti~
~ s ~ s D; r~ r p4 ~P2 Another alternative is to generate a carbene from a dihaloalkane or trihalo~ U la. Ie with a base in the pr~sence of the olefin (Von E. Doering W. et a/. J.
Am. Chem. Soc. 76, 6162 (1954)) followed by dehalogenation, for example accon~ing 5 to Jefford C. W. et a/. J. Am. Chem. Soc. 94, 8905 (1972). Cycloaddition between diazomethane and compounds of formula 17 provides a pyrazoline intermediate which upon photolysis and deprotection prodl Ices spirocyclopropane 18 (Samano V.
ef al. Tefrahedron Leff. 35(21), 3445 ~1994)). The deprotection of the anomeric center is in tum accon l,~lished using one of the many procedures well known to those 10 skilled in the art, e.g. Greene, T. W., Protective Groups in Organic Chemistry, John Wiley&Sons, NewYork, (1981).
Carbohydrates of formula 20 are made by a wide variety of procedures.
Reaction of olefin 17 with ketene under the conditions of Redlich H. ef al. Angew.
Chem. 101 (6), 764 (1989) gives cyclobutanone 19 (Scheme 6), with is then 15 deoxygenated using the procedure of Mori K ef al. Tefrahedron, 43(10), 2229 (1987) or Romming C. et al. Acta Chem. Scan. B, 40(6), 434 (1986). The free reducing sugar is then obtained as mentioned above (Greene T. W. Profecfive Groups in Organic Chemistry John Wiley & Sons, New York, 1981).

~OCH3 O o~_OCH3 ~ OH
's ~ , . Dooxy~,-h~tiùn 0~
P~O oP2 ll P~Ci OP2 D~,ut~un p,~ ~P2 W O 96/40705 PCT~US96/10404 An alternative route uses the photochemical cyclization of a ca, bol ,ydrate of forrnula 22 to give cyclobutanol 23 (Schenne 7, Paquette L. A. ef al.
J. Am. Chem. Soc. 108(13), 3841 (1986). Deoxygenalion of alcohol 23 occurs accordi,~g to the procedure of Barton D. H. R. ef a/. Pure Appl. Chem. 53, 15 (1981).
5 Precursor 22 is made by 4-alkylation of the corresponding aldehyde derived from selectively protected methyl riboside 21, e.g. Youssefyeh R. D. ef a/. J. Org Chem.
44(8), 1301 (1979).

H OH
H0~ OC H3 ~~O~C~ ~o~ /\~O OH
~!/ r H3C \_/ hv \/\J D~o~ ~
P,~ ~P2 P~ ~)P2 P''5 ~P2 FP~ ~P2 21 ~ 23 2D

Alkylation of aldehyde 15 with a two carbon dielectrophile, preferably 10 diiodoethane, gives 4~isl Ihstituted aldehyde 24 (Scheme 8, Youssefyeh R. D. ef al.
J. Org Chem. 44(8), 1301 (1979)). Treatment of aldehyde 24 with a metal or metalsalt, preferably samarium diiodide (Molander G. A. ef al. J. Am. Chem. Soc. 109(2), 453 (1987)), or with an organometallic reagent, preferably an alkyllithium (Vanderdoes T. ef al. Tefrahedron Left 27(4), 519 (1986)), achieves the ring closure.
15 The cyclobutanol is then deoxygenated and deprotected at the anomeric position using the procedures previously mentioned to provide spirocyclobutylfuranose 20.
o O OH
H'~,~O OR Ba~ H~,o OR M ~~ OR ~ o OH
~Y ~L ~ R-L~ \~ b~ Ay~ ~dbOr~ ~
P,~i OP2 P~i OP2 P,d OP2 P,~i OP2 An altemative method to make carbohydrate of formula 20 is illustrated in Scheme 9. ~ctivation of the two primary hydroxyl groups using the many 20 procedures well known to those skilled in the art (Laroc:k R. C. Comprehensive =

W096/40705 PCT~S96/1040~s Organic Transformafsions, VCH Pubiishers, Inc. New York (1989)) followed by diallylation with a malonate produces dicarboxyiate spirocyclobutane 27 (PecquetP. et al. Heterocycles 34(4), 739 (1992)). Decarboxylation using the procedure of Tufariello J. J. ef al. Tefrahedron Lett. 6145 (1966) followed by deoxygenation and 5 de,crotection of cyclobutanone 19 as previously mentioned gives spirocyclobutane 20.
Carbohydrate of formula 29 iS made by activation of one of the primary hydroxyl groups of diol 25 (Scheme 10, Larock R. C. Comprehensive Organic Transformafions, VCH Publishers, Inc. New York, (1989)). Cyclization occurs upon10 treatment of alcohol 28 with a base (Koll P. et al. Angew. Chem. Inf. Ed. Engl. 25, 368 (1986)). The anomeric position is then deprotected as previously mentioned to afford spirooxetanofuranose 29.
O O
HO O ~R L O ~R RO/~OR R ~ ~R
X~ ~ BaSe P~ OP2P,~5 OP2 D~ o~OR D~ XJ~ ~dSOn O~~H
s s Deprul~cli~ n P~ OP2 P,~ OP2 Alternatively, compound 29 iS obtained via Mitsunobu reaction of diol 15 25 under the conditions of Berkowitz W. F. et al. J. Org. Chem. 52(6), 1 1 19 (1987) which gives compound 29 after de,u, olec~ison. As another alle, I ,~Li~e, lithium chloride treatment of a cyclic carbonate derived from diol 25 followed by deprotection also gives carbohydrate 29.

CA 02220642 l997-ll-28 W O 96/40705 PCT~US96/10404 HO~ OR HX~OR Base ~, ii s D~ q P~ OP2 P,t5 '~P p,t5 ~P2 The syntheses of the carbohydrates of fonnula 32 are illustrated in Schemes 11, 12 and 13. Treatment of activated carbohydrate 31 with a base prior to deprule~;lio~ I gives spiroa~e~idinofuranose 32 (Scheme 11, Vaughan W. R. et a/.
5 J. Org. Chem. 26, 138 (1961)). Aminoalcohol 30 is prepared following the procedures used to make carbohydrates of formula 2 (Scheme 3).

RHNX~ RHN~i D~p. ~ n s P,p OP2 P,~ OP2 p,p ~P2 Alternatively, treatment of diactivated compound 26 with ~" "no"ia, a primary amine, a protected amine or an activated amine (Scheme 12) followed by 10 deprotectiongivesc ,I,ohydrate32. See, Juaristi E. etal. 7etrahedron Lett. 25(33), 3521 (1984).

LX~ R-NH2 RN~
P~p OP2 D~ P~ P2 26 ~

An other alternative is to treat azidoalcohol 33 with a trialkyl or triaryl phospl ,ine (Scheme 13, Szmuszkovicz J. et al. J. Org. Chem. 46(17), 3562 (1981)).

15 Decomposition of the azido group and Mitsunobu like cyclization gives after deprotection azetidine 32.

W O 96140705 PCT~US96/10404 N N~ ~ . PR3 2 ~ D~,u~i~n ~ ~
P,p ~)P2 P,t5 ~)P2 (B) PREPARATION OF THE HETEROCYCLE
Heterocycles for compounds of formula 1 (Scheme 1 ) where Y=C, F' and D' are s~ Ihstit~ ItPd aroma~ic groups, pr~r~, ~bly para-fluorophenyl, G is hydrogen, 5 and E' is a hydl u~en, aikyl, prt:rer~bly hydl uyen, are made via a pyrrole intermediate 37 made using the procedure of Gewald, K. Z Chem., 1, 349 (1961). Alternatively,37 is made by condensing phenone 34, where L is halide or sulfonate, with phthalimide 35 in order to introduce the pyrrole nitrogen. Knoevenagel condensation of ketone 36 with malonitrile followed by removal of the phthalimide protecting group 10 affords pyrrole 37.
Upon treatment with an orthoester, preferably triethylorthoformate, an imidate is formed which is further condensed with a substituted aniline, preferably para-fluroaniline, to give diaryl-pyrrolopyrimidine 39 (Taylor, E. C. et a/. J. Am Chem.
Soc. 87(9),1995 (1965)). Additionally, the pyrrolopyrimidine can be further 15 functionalized at the 6 position, when E' is methyl, by treatment with N-b~ u, "osuccinimide (Saroja ,B. et al. Tetrahedron Lett 1984, 25(47), 5429). Treatment of this bl on ,on ,ethylene with a nucleophile or with an alkyllithium and an electrophile allows easy introduction of functional groups such as amino or guanidino.

WO 96/40705 PCT~US96/10404 Rl 0 ~ B R2 ~N~

R3 5 R3 ~R5 3~ 35 3~S

gN R h--~_R5 (EtO)3CH R~ ~.~R5 R2~ jCN R2. ~ CN
2) CH30Na E'~-- E'~--N~--OEt 3~ 38 R'2 R.~N~ R5 R'2 R'1 ~ R1 SC5~EME 14 Heterocycles for compounds of formula 1 (Scheme 1 ) where Y=N F
andD aresubstitutedaro",alicgroups preferablypara-fluorophenyl.

> ~ R~ 1)5~ pholin~l ~ 1)foma~nide NC 2) NH 2NH 2 H2 /N 2) POC13 " '~

Ra ~-NH ~2 h 42 SCHEME 1 ~
Compounds where E is "nothingn are made using the procedure of Kobayashi, S.
Chem Pharm. Bull. (JapanJ 21, 941 (1973). ~noevenagel condens~LiGn of ",alonil,ile 5 with a substituted benzaldehyde, preferably para-fluorobenzaldehyde, followed by treatment with h~d~ ~ine gives 5-aminopyrazole~-carbonitrile 40 (Scheme 15). The4-chloro-pyrazolo[3,4-d]pyrimidine 41 is obtained upon ring closure reaction with ru""~",ide and chlorination using the procedure described by Cheng, C. C. J. Org.
Chem. 21, 1240 (1966). Treatment of chloride 41 with ammonia, for the 4-amino 10 series, or a sl ~hstit~ Ited aniline preferably para-fluoroaniline, as previously mentioned gives diarylpyrazolopyrimidine 42.

C. COUPLING OF THE CARBOHYDRATE WITH THE HETEROCYCLE
The coupling of the carbohydrate 2 with pyrrolo~2,3-d]pyrimidine heterocycles is acco",plished as follows (Scheme 16). The sugar is first converted 15 to its 1-halo derivative, preferably chloro, by reacting it with CC14 and HMPT by a procedure described in Wilcox, C. T. eta/. Tetrahedron Leff. 27(9), 1011 (1986). The halo derivative is condensed with the anion of the heterocycie 3 (where Y is carbon and E is h~rdl ugen) using a phase ~ ~"arer catalyst such as TDA-1. Rosemeyer H., and Seela, F, Helvetica Chimica Acta, 71:1573 (~1988,). The resulting blocked nuclsosides are deprotected by a variety of procedures well known to those skilled in the art. F' B' O ~ ~ ~E' ActivatiolD B~o oP! ~~z 2 ) Coupling _ ~
2 ~PI ~P z Coupling of sugars to the pyrazolo[3,4-d]pyrimidine bases is performed by Lewis acid catalysis conditions. Cottom, et al., J. Med Chem.. 27, 11210 (1984). In such cases the sugars are converted to their 1-O-acyl form, preferably 1-O-acetyl, by again using one of the many standard acetylation 10 ptucedures. A mixture of the heterocycle 3 (where Y is nitrogen) and the acetylated sugar in boiling nitromethane is treated with boron trifluoride diethyl etherate. The products are purified by chromatography or crystallization, and are cle,urolected to obtain the final compounds.

D. MODIFICATION OF SUBSTITUENTS ON THE HETEROCYCLE
Due to the chemical incompatibility between some of the substituent on the heterocycle and the glycosidation-reaction conditions, the final fi~ ;Lio,~li7~fion of the nucleoside is done after the coupling reaction. For example, the 5-aryl group is introd~ Ir~3d onto the pyrrolopy, il "idir ,e ring system using one of the many palladium-catalyzed cross coupling procedures (review: Stille, J. K. Ang.
20 Chem., In~. Ed. Engl. 25, 508(1986)).
Typically, a 4-substituted-amino-5-halopyrrolo[2,3-d]pyrimidine 44, where the halogen is iodo, is coupled to an arylbororsic acid (i.e. A=B(OH)2 in Scheme 17) in the presence of a catalyst such as tetrakistriphenylphosphine p~ll~rlil Irr~.

R~2 R~4 R'~ R~3 E' 43 44 Su3ar R~
R~4 R~5 R~3 R~3~A R4~ R'5~R,2 Pd~ ~N~NJ~E' Sugar Altematively, in place of aryl boronic acids, other activated aryl co",pounds such as aryltrialkyltin(A=Sn(alkyl)3 is successfully used to obtain the final product 45.
5 S~ ~hstih ~tion of the aryltrialkyltin by an unsaturated trialkyl stannane by procedures as described but not limited to the one described in Stille, J. K. Ang. Chem.. Int. Ed.
Engl. 25, 508 (1986) provides the 5-alkenyl derivative which can be hydrogenated in order to prepare the corresponding alkyl analog.
Further modifications can be added to the aromaLic rings after cross 10 coupling with the heterocycle either before or after gl~,~;osid~lion. Reduction, oxidation and/or deprotection steps are done at this stage. For instance a cyano group is oxidized to its carboxamide or reduced to its amine. A N-phenylacelamide is depr~,lected and kept as its aniline or l,~n~r~""ed to its trifluoromethanesulfonamide to improve water solubility.

WO 96/40705 PCT~US96/10404 R N-U
X (lH2)n (l~)n R2~ chain extension ~/ aminaUon ~/R4 R1~\R5 lljllu~u.~ 'I R~\R5 R~R5 Base Base Base 4~ 46R=OH 48 47 R=L

Carbon chain extension is done at this stage (Scheme 18). The aromatic ring substituent on the glycosiiated intermediate 45 where X is halide or trifluoru" ,e~h~l ,esulfonate is coupled with a vinyl or allyl trialkyltin species using one 5 of the many palladium-catalyzed cross coupling procedures ~review: Stille supra).
The double bond is then oxygenc~led at the terminal ,~)osi~io, ! and the resulting alcohol 46 is converted to a leaving group L ~rer~r~bly iodide (Srivastava P. C. et al. J. Med Chem.1975 18(12) 1237). Displ~cement by an amine cornpletes the carbon chain extension and improves the water solubility for compounds of formula 48.
Other ~o~"lial water solubilizing groups such as guanidino derivatives can be ~3, epar~:d from the corresponding amino or hydroxy compounds by application of methods described in the literature such as but not limited to the proceduresdescribed by Miller and Bischoff (Synfhesis 778 (1986)) Dodd and Kozikowski (Tefrahedron Leff. 35 977 (1994)) Beatty and Magrath (J. Chem. Soc. 12 (1965)) 15 Larson et al (Inf. J. Pepf. Profe~n Res. 9, 182(1977)) Brancl and Brand (Org. Synth.
22 59 (1942)) Ichikawa (Tefrahedron Leff 29 4957(1988)) Katritzky et al (Synfh.
Commun. 25 1173 (1995)) Ariga and Anslyn (J.Org. Chern. 57 417 (1992)) Palat et al (Collecf. Czech. Chem. Comm~m. 57 1127 (1992)) or M. S. Bernatowicz (J.
Or~. Chem. 57 2497 (1992). Acylguanidines can be ~ par~d by methods described 20 in the literature such as the methods described by Bock et al (J. Med.Chem. 29 1~40 (1986)) and lefere-lces cited therein.

W 096/40705 PCT~US96/10404 E. REMOVAL OF THE PROTECTING GROUPS
Acid labile protecting groups such as ketals, silyl ethers or ethers are removed using a dilute acid or a weak organic acid, e.g. 0.1 N hydrochloric acid or 70% aqueous trifluoroacetic acid (Greene, T. W., Protective Groups in Organic 5 Ci ,e" ,isl, ~ John Wiley & Sons, New York (1981). Base labile protecting groups such as acyls or ca, LJa" IGleS are removed by ll e~ enl with an organic or i"or~al ,ic base, e.g. sodium ,nell ,uxide, sodium hydroxide, ammonia (Id). Benzyl protec~i"g groups are removed by hydrogenolysis in the presence of a metal catalyst, preferably palladium chloride. Shen, T. Y. eta/. J. Org. Chem. 30, 835 (1965).
Preferred compounds of the invention, which can be made using the methods described, include the following.

EXAMPLES
Example 1 Preparation of comPound of formula 10 2.3.5-Tri-O-(phenylmethyl)-1 -(1,3-dithian-2-yl)-D-nbo-pentane Boron trifluoride diethyl etherate (11.4 mL, 92.4 mmol) was added to a solution of methyl 2,3,5-tri-O-(phenylmethyl)-D-ribofuranoside (30 9, 66 mmol)(Barker, R. and Fletcher, H. G. J. Org. Chem.1961, 26,4605) and 1,3-propanedithiol (10 mL, 99 mmol) in dry dichloromethane (130 mL) at 48~ C. The reaction mixture was stirred 30 minutes at -48~ C and warmed to room temperature in the course of20 one hour. After stirring at room temperature for one hour, the mixture was quenched with saturated aqueous sodium bicarbonate, diluted with ethyl acetate and washedwith saturated ~queo~ ~-s sodium bicarbonate and saturated ~ eo~ ~s sodium chloride.
The organic layer was dried over sodium sulfate and conce, ILI aled under reduced pressure. The residue was purified by flash chromatography on silica gel 25 (hexanes/ethyl acetate 90/10 to 75/25). Yield 31.9 9, 94 %, Rf = 0.3 (silica gel, hexanes/ethyl acetate 80/20).

WO 96/40705 PCT~US96/10404 Example 2 Pl ~paralion of compound of formula 11 (3S. 4R)-1.3.4-Tri-[(phenylmethyl)oxy~-5-(1.3-dithian-2-yl)Dentan-2-one A solution of dimethyl sulfoxide (22.1 rnL, 312 mmol) in dry dichloromethane (100 mL) was added dropwise over 10 minutes to a solution of 5 oxalyl c hlolide (16.3 mL,187 mmol) in dry dichlo,u"~ethane (200 mL) at -78~ C. After stirring 10 minutes at-78~C, a solution of the compound of Example 1 (31.9 g, 62.4 mmol) in dry ~Jicl .l ~ ro",~Ll ,a, le (100 mL) was added dropwise to the reaction mixture over 20 minutes at -78~C. After stirring at -78~C for 2() minutes, a solution oftriethylamine (87 mL, 624 mmol) in dry dichloromethane (100 mL) was added 10 dropwise over 10 minutes at -78~C. Affer completion of the addition the internal temperature was allowed to raise to -40~ C over 30 minutes. The reaction mixturewas quenched with saturated aqueo~s ammonium chloride and warmed to room te"~er~lure. The layers were separated and the ~q~ ~eo~ ~s layer was back extracted twice with cJichloromeU ,~ne. The combined organic extracts were dried over sodium 15 sulfate and cc ~ Icenl~ aLecl under redl ~c d pressure. The residue was purified by flash ~ llumaLuylap hy on silica gel (hexanes/ethyl acetate 90/10 l:o 70/30). Yield: 27.9 g, 88 %, Rf = 0.35 (silica gel, hexanes/ethyl acetate 80/20).

Example 3 Pre~aration of compound of formula 12 ~C-[(Phenylmethyl)oxy]methyl-2.3.5-tri-0-~henYlmethyl-1 -(1,3-dithian-2-yl)-D-ribo-20 pentane.
A solution of the compound of Example 2 (1 g, 1.97 mmol) in drytetrahydrofuran (25 mL) was added dropwise over 5 minutes to a solution of [(phenylmethyl)oxy~methyllithium ~3.94 mmol) (Still, W. C. J. Am. Chem. Soc. 1978, 100, 1481) in dry tetrahydrofuran (25 mL) at -78~ C. After stirring for 20 minutes at -25 78~C, the reaction mixture was quenched with saturated ~ql~eo~s ammoniumchloride, warmed to room temperature, diluted with ethyl acetate and washed with saturated aqueous ammonium chloride. The organic layer was dried over sodium sulfate and concer,L, ~led under reduced pressure. The residue was purified by flash ch, c" "~ography on silica gel (hexanes/ethyl acetate 90/10 to 75/25). Yield: 1.05 g, 30 85 %, Rf = 0.45 (silica gel, hexanes/ethyl acetate 70/30).

Example 4 PreParation of compound of formula 2 4-C-[(Phenylmethyl)oxy]methyl-2.3.5-tri-O-(phenylmethyl)-D-ribofu, c, ,ose A heterogeneous mixture of calcium carbonate (4 9, 40 mmol), jGdUIIIe~ l ,e (1.25 mL,20 mmol) and the compound of Example 3 (2.52 9, 4 mmol) 5 in acetonitrile/tetrahydrofuran/water (1/1/9, 44 mL) was refluxed overnight (Fetizon, M. J. Chem. Soc., Chem. Comm. 1972,382). More iodomethane (1.25 mL, 20 mmol) was added and refluxing was pursued for 24 hours. The mixture was cooled, diluted with ethyl ~cel~le and washed with saturated aq~ ~eo~ ~s sodium chloride. The ~q~ ~eo~ ~s layer was extracted with dichloro"~elhal ,e and the combined organic extracts were 10 dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (hexanes/ethyl ~cet~te 80/20 to 65/35). Yield: 2.06 9, 95 %, Rf = 0.2 (silica gel, hexanes/ethyl acetate 70/30).
Example 5 Preparation of compound of formula 4 4-N-Phenylamino-5-phenyl-7-(4-C-~(phenylmethyl)oxy]methyl-2.3.5-tri-O-15 (phenylmethyl)-B-D-ribofuranosyl)pyrrolo[2.3-dlPyrimidine Hexamethylphosphorous triamide (415 ,uL, 1.95 mmol) was added to a solution of carbon tetrachloride (250,uL, 2.6 mmol) and the compound of Example 4 (349 mg, 0.65 mmol) in dry toluene at -78~ C. The reaction mixture was warmed to 0~ C in the course of one hour and stirred at 0~ C for 30 minutes. The orange solution 20 was quenched with water, diluted with toluene and washed with water and saturated aqueous sodium chloride. The organic layer was dried over sodium sulfate and concentrated under red~ ~ce:l pressure to a volume of c.a. 5 mL. The chloro-sugar solution was added to a mixture of 4-N-phenylamino-5-phenyl-pyrrolo[2,3-~pyrimidine (370 mg, 1.3 mmol), finely powdered potassium hydroxide (85%,170 25 mg, 2.6 mmol), tris[2-(2-methoxyethoxy)ethyl]amine (420 ~L, 1.3 mmol) and 4 Amolecular sieves in dry toluene which had been stirring at room temperature for 2 hours. After stirring ovemight at room temperature, the reaction mixture was filtered through Celite6~ and the filtering pad was rinsed with ethyl acetate. The filtrate was diluted with ethyl ~ e and washed with saturated aqueous sodium chloride. The 30 organic layer was dried over sodium sulfate and conce"l,aLed under reduced pressure. The residue was purified by flash chron,aLography on silica gel W O 96140705 PCT~US96/10404 (hexanes/ethyl ~cet~te 90/10 to 70/30). Yield: 229 mg, 44 %, Rf = 0.6 (silica gel, hexanes/ethyl ~cePte 80/20).

Example 6 Preparation of compound of formula 1 4-N-Phenylamino-5-Phenyl-7-(4-C-hydroxymethyl-B-D-ribofuranosyl)pyrrolo~2.3-5 dlPyrimidine: Table 1 #1 A mixture of palladium hydroxide (200 mg) ;and 4-N-phenylamino-5-phenyl-7-(4-C-[(phenylmethyl)oxy]methyl-2,3,5-tri-O-(phenylmethyl)-~-D-ribofuranosyl)pyrrolo~2,3~pyrimidine (212 mg, 0.26 mmol) in acetic acid/ methanol(1/1, 10 mL) was vigorously stirred at room temperature under one atmosphere of10 hy~, ogel ,. After 7 days of stirring the reaction mixture was filtered through Celite~
and the filtering pad was rinsed with hot methanol. The filtrate was cGncenl,a~ed under reduced pressure and the solid residue was recrystallized from ethanol. Yield:
30 mg, 25 %, Rf = 0.4 (silica gel, dichloromethane/methanol 80/20), mp 232~ C.

Example 7 Preparation of compound of formula 10 15 5-Deoxy-2.3-di-O-(phenylmethyl)-1 -(1.3-dithian-2-yl)-D-nbo-pentane The title compound was synthesized following a procedure analogous to the synthesis described in Example 1. Thus methyl 5-deoxy-2,3-di-O-(phenylmethyl)~-ribofil, ~noside (7 g,21.3 mmol) prepared ~y a procedure an~logo~ l~
to the synthesis of methyl 2,3,5-tri-O-(phenylmethyl)-r:)-ribofuranoside (Barker, R. and 20 Fletcher, H. G. J. Org. Chem. 1961, 26,4605), gave 7.9 9,92 %, Rf = 0.35 (silica gel, hexanes/ethyl acetate 70/30).

Example 8 Preparation of compound of formula 11 (3S.4R)-3 4-Bis-~(phenylmethyl)oxy~-5-(1 3-dithian-2-yl)pentan-2-one The title compound was synthesized following a procedure analogous 25 to the synthesis described in Example 2. Thus the compound of Example 7 (7.9 g, 19.5 mmol) gave 6.32 9, 80 %, Rf = 0.2 (silica gel, hexanes/ethyl acetate 70/30).

W O 96/40705 PCT~US96/10404 ExamPle 9 Preparation of compound of formula 12 5-Deoxy-4-C-methyl-2.3-di-O-(phenylmethyl)-1-(1 3-dithian-2-yl)-D-nbo-penlane - A solution of the compound of Example 8 (2 9, 5 mmol) in dry tetrahydrofuran (30 mL) was added dropwise over 10 minutes to a solution of 5 methyllithium (20 mmol) in dry tetrahydrofuran (20 mL) at -78~ C. After stirring for 20 minutes at -78~C, the reaction mixture was quenched by slow addition of a solution of acetic acid (2 mL) in dry tetrahydrofuran (10 mL) over 5 minutes at -78~C. The quenched solution was warmed to room temperature, diluted with ethyl acetate andwashed with saturated ~queous sodium bicarbonate and saturated aqueous sodium 10 chloride. The organic layer was dried over sodium sulfate and concel ILI aled under reduced pressure. The residue was purified by flash chromatography on siiica gel(hexanes/ethyl acetaLe 85/15 to 75/25). Yield: 2.038g, 98 %, Rf = 0.38 (silica gel, hexanes/ethyl ~cet~te 70/30).

Example 10 Preparation of compound of formula 2 15 5-Deoxy~-C-methyl-2-3-di-O-(phenylmethyl)-D-ribofuranose The title compound was synthesized following a procedure analogous to the synthesis described in Example 4. Thus, the compound of Example 9 (2.04 g, 4.87 mmol) gave 1.4 g, 88 %, Rf = 0.4 (silica gel, hexanes/ethyl acetate 70/30).
Example 11 PreParation of compound of formula 2 20 5-Deoxy-4-C-methyl-2.3-O-(methylethylidene)-D-ribofuranose A mixture of p~ rn hydroxide (0.5 9) and the compound of Example 10 (2.62 g,7.98 mmol) was vigorously stirred at room temperature for 3 hours under one atmosphere of hydrogen. The reaction mixture was filtered through Celite~ and the filtering pad was rinsed with hot methanol. The filtrate was concenll dled under 25 reduced pressure and azeotroped twice with dimethylformamide. The residue wasdissolved in dimethylformamide (10 mL). p-Toluenesulfonic acid monohydrate (catalytic) and 2,2-dimethoxypropane (4.6 mL, 32 mmol) were added. After stirring overnight at room temperature, the reaction mixture was diluted with ethyl acetate and washed with saturated aqueous sodium bicarbonate and saturated aqueous 30 sodium chloride. The Ol gar,ic layer was dried over sodium sulfate and concentrated W O 96/4070~ PCT~US96/10404 under reciuc~d pressure. The residue was purified by flash chromaloyl c~p hy on silic,a gel (hexanes/ethyl ~cel~l~ 80/20 to 70/30). Yield: 507 mg, ~4 %, Rf = 0.3 (silica gel, hexanes/ethyl ~cet~te 70/30).

Example 12 Preparation of compound of formula 4 5 4-N-Phenylamino-5-phenyl-7-(5-deoxy4-C-methyl-2.3-O-(methylethylidene)-~-D-ribofuranosyl)pyrrolo-~2.3-dlPyrimidine Hexamethylphosphorous triamide (800 ~L, 4.35 mmol) was added to a solution of carbon tetrachloride (600,uL, 5.8 mmol) and the compound of example 11 (272 mg,1.45 mmol) in dry toluene at -50~ C. The reacltion mixture was warmed10 to -10~ C in the course of 30 minutes and stirred at -10~ C for 15 minutes. The orange solution was quenched with water, diluted with toluene anci washed with water and saturated ~q~ ~eous sodium chloride. The oryanic layer was ciried over sodium sulfate and cc "ce"~ aled under red~ ~ced pressure to a volume of C.cl. 5 mL. The chloro-sugar solution was added to a mixture of 4-N-phenyiamino-5-phenylpyl ,~ 3~pyrimidine 15 (830 mg,2.9 mmol), finely powdered po~C5jllrll hydroxide (85%, 380 mg, 5.8 mmol) and tris[2-(2-r"~U ,oxyethoxy)ethyl]amine (925,uL,2.9 mmol) in dry toluene which had been stirring at room temperature for 90 minutes. After stirring overnight at room temperature, the reaction mixture was diluted with ethyl -~cet~le and washed with saturated aq~ ~eo~ ~s ammonium chloride. The organic laye!r was dried over sodium 20 sulfate and conce"l, clled under reduced pressure. The residue was purified by flash chr~m~lography on silica gel (hexanes/ethyl acelale 70130 to 50/50). Yield: 223 mg, 34 %, Rf = 0.3 (silica gel, hexanes/ethyl acetate 60/40).

Example 13 Prer~aration of compound of formula 1 4-N-Phenylamino-5-phenyl-7-(5-deoxy4-C-methyl-~-D-ribofuranosyl)pyrrolo~2.3-25 dlpyrimidine A solution of 4-N-phenylamino-5-phenyl-7-(5~ieoxy4-C-methyl-2,3-O-(methylethylidene)-,B-D-ribofuranosyl)pyrrolo[2,3-dlpyrimidine (220 mg) in 70 %
aqueous trifluoroacetic acid (20 mL) was stirred at 0~C for one hour and at roomtemperature for one hour. The reaction mixture was conc,entrated under reduced 30 pressure and azeotroped twice with water and twice with ethanol. The residue was CA 02220642 1997-ll-28 W O 96/40705 PCT~US96/10404 neutralized with saturated aqueous sodium bicarbonate and the precipitated nucleoside was filtered and rinsed with water. The solid was recovered and - recrystallized from ethanol. Yield: 130 mg 65 % Rf = 0.5 (silica gel dichloromethane/methanol 90/10) mp 198-200~ C.
-5 Example 14 Prd,~a, ~lion of co" ,pound of formula 18 Methyl 2.3-O-(methylethylidene)-4-C-spirocyclopropvl-D-erythrofuranoside A solution of methyl 5-deoxy-2 3-O-(methylethylidene)-~-D-erythr~
pent4-enofu,anoside (2 g 10.7 mmol) (Inokawa S. et a/. Carbohyd. Res. 1973, 30, 127~ and diodor,)t:LI ,ane in dry ether (20 mL) was added dropwise over 4 hours to a 10 refluxing suspe"siori of freshly made zinc-copper couple in dry ether. The reaction mKture was refluxed ove" .:~I ,I cooled diluted with ether and washed with saturated ~ql ~eol Is a" " "o" - lm chloride. The organic layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (pentane/ether 90/10 to 80/20) to provide the title15 compound 18 (1 9 47 %) Rf = 0.3 (silica gel hexanes/ethyl ~cet~te 90/10).

Example 15 Preparation of compound of formula 18 2.3-o-(Methylethylidene~-4-c-spirocyclopropyl-D-erythrofuranose A mixture of methyl 2 3-O-(methylethylidene)-4-C-spirocyclopropyl-D-erythrofuranoside (2.57 9 12.8 mmol) 1 N aqueous hydrochloric acid (20 mL) and 20 tetrahydrofuran (20 mL) was refluxed for 1 hour. The cooled reaction mixture was neutralized with DOWEX~91X8-200 ion exchange resin (OH-form) filtered and rinsedwith methanol. The combined filtrates were concentrated under reduced pressure and azeotroped twice with dimethylformamide. The residue was dissolved in dimethyl~ ar"ide (10 mL). ~Toluenesulfonic acid " ,onol "~drate (catalytic) and 2 2-2~ dimethoxypropane (4.6 mL 32 mmol) were added. After stirring 4 hours at roomtemperature the reaction mixture was diluted with ether and washed with saturatedaqueous sodium bicarbonate and saturated aqueous sodium chloride. The organic layer was dried over sodium sulfate and concentrated under reduced pressure. Theresidue was purified by flash chromatography on silica gel (pentane/ether 70/30 to 30 40/60). Yield: 1.2 9 50 % Rf = 0.4 (silica gel hexanes/ethyl acetate 60/40).

W O 96/40705 PCTnUS96/10404 Example 16 Pr~aralion of compound of formula 4 4-N-(4-Fluorophenyl)amino-5-phenyl-7-(2.3-O-(methvlethylidene)-4-C-spirocyclopropyl-~-D-erythrofuranosyl)pyrrolo[2.3-dlPyrimidine The title compound was synthesized following a procedure analogous 5 to the synthesis desuibed in example 12. Thus coupling of 2,:3-O~methylethylidene)-4-Gspirocyclopropyl-D-erythrofuranose (450 mg, 2.42 mmol) with 4-N-(4-fluorophenyl)amino-5-phenylpyrrolol2,3-~pyrimidine (20, 1.47 9, 4.84 mmol) provided the title nucleoside (294 mg, 26 %), Rf = 0.6 (sllica gel, hexanes/ethyl acetaLe 70/30).

10 Example 17 Preparation of compound of formula 1 4-N-(4-Fluorophenvl)amino-5-phenvl-7-(4-C-spirocyclopropyl-~-D-erythrofuranosyl)pyrrolo~2.3-dl~yrimidine:Table4 ~150 The title compound was s~- ,Ihesi~ed following a procedure analogous to the synthesis desuibed in Example 13. Thus 4-N~4-fluorophenyl)amino-5-phenyl-15 7~2,3-O-(methylethylidene)4-C-spirocyclopropyl-~-D-erythrofuranosyl)pyrrolo[2,3-o~pyrimidine (289 mg, 0.6 mmol) provided the titled depn,~ected nucleoside (159 mg, 60 ~/0), Rf = 0.5 (silica gel, dichloromethane/methanol 90/10), m.p.142~C.
Example 18 Preparation of comPound of formula 4 4-chloro-5-iodo-7-(5-deoxy-4-c-methyl-2.3-o-(methylethylidene)-B-D
20 ribofuranosyl)pyrrolo~2 3-dlpyrimidine The title compound was synthesized following a procedure analogous to the synthesis desuibed in Example 12. Thus coupling of 5-deoxy4-C-methyl-2,3-O-(methylethylidene)-D-ribofuranose (550 mg, 2.9 mmol) with 4-chloro-5-iodo-pyrroloE2,3~pyrimidine (1.23 9, 4.35 mmol) provided the titled nucleoside (581 mg, 25 44 %), Rf = 0.4 (silica gel, hexanes/ethyl acetate 60/40).

Example 19 r, eparalion of cor"~ound of formula 1 4-Chloro-5-iodo-7-(5-deoxy-4-C-methyl-~-D-ribofuranosyl)pyrrolo~2,3-~Pyrimidine ~ The title compound was synthesized following a procedure analogous to the synthesis described in Example 13. Thus 4-chloro-5-iodo-7-(5-deoxy4-C-5 methyl-2,3-O~(methylethylidene)-~-D-ribofuranosyl)pyrrolo-~2,3~pyrimidine (100 mg, 0.22 mmol) provided the titled depr~ected n~ ~cleoside (14 mg,15 %), Rf = 0.45 (silica gel, dichloromethane/methanol 90/10), m.p. 173-174~ C.

Example 20 Preparation of compound of formula 4 4-chloro-5-iodo-7-(2~3-o-(methylethylidene)-4-c-spirocyclopropyl-B-D
1 0 erythrofuranosvl)pyrrolo~2.3-dlPyrimidine The title compound was synthesized following a procedure analogous to the s)/r,U,esis described in Example 12. Thus coupling of 2,3-O~methylethylidene)-~spirocyclopropyl-D-erythrofuranose (500 mg, 2.66 mmol) with 4-chloro-5-iodo-pyrrolo[2,3~pyrimidine (1.11 9, 3.99 mmol) provided the titled nucleoside (402 mg, 15 34 %), Rf = 0.7 (silica gel, hexanes/ethyl acetate 70/30).

Example 21 Preparation of compound of formula 1 4-Amino-5-iodo-7-(4-C-spirocyclopropyl-B-D-erythrofuranosvl)pyrrolo~2.3-diPvrimidine: Table 4 ~175 Liquid ~"",lu,.ia (15 mL) was added to a solution of 4-chloro-5-iodo-7-20 (2,3-O-(methylethylidene)-4-C-spirocyclopropyl-~-D-erythrofuranosyl)pyrrolo~2,3-d~pyrimidine (200 mg, 0.45 mmol) in methanol (15 mL) at -78~C. The reaction mixture was heated at 100~ C in a sealed steel bomb for 24 hours. Ammonia was slowly released from the cooled bomb and the resulting solution was conce, Ill aLed under r~duced pressure. The residue was disolved in 70 % aqueous trifluoroacetic25 acid and stirred at room temperature. After 30 minutes the reaction mixture was conce, ILI aLed under reduced pressure and azeotroped twice with water and twice with ethanol. The residue was neutralized with saturated aqueous sodium bicarbonate and the precipitated nucleoside was filtered and rinsed with water. The solid was recovered and recryst~ili7~d from ethanol. Yield: 73 mg, 42 %, Rf = 0.35 (silica gel, 30 dichloromethane/methanol 90/10), m.p. 232~C (dec).

_ W O 96/40705 PCT~US96/10404 Example 22 Preparation of compound of formula 12 4-C-Methyl-2.3.5-tri-O-(phenylmethyl~-1 -(1.3~ithian-2-yl)-D-/yxo-pentane The title compound was synthesi7ed following a procedure analogous to the synthesis desuil,ed in Example 9. Thus the compound of Example 2 (5 9 9.85 mmol) gave the title compound (3.94 9 84 %) from a separ~ble 12/1 epimeric mixture Rf = 0.38 (silica gel hexanes/ethyl acetate 70/30).

ExamDle 23 Preparation of comDound of formula 2 4-c-Methvl-2.3.5-tri-o-(phenylmethyl)-D-lyxofuranose The title compound was s~, lll ,esi~ed following a procedure analogous 10 to the synthesis described in Example 4. Thus the compound of Example 22 (3.94 9 7.51 mmol) gave the title compound (2.6 9 80 %) Rf = 0.25 (silica gel hexanes/ethyl acetate 70/30).

ExamPle 24 Pl l~udl ~lion of compound of formula 4 4-N-~4-rDimethylaminomethyl)Phenyl3amino-5-~henyl-7-(4-C-methyl-2.3,5-tri-0-15 (phenylmethyl)-B-D-lyxofuranosyl)pyrrolo~2 3-dlryrimidine The title compound was synthesi~Pd following a procedure analogous to the synthesis described in Example 12. Thus coupling ol 4-C-methyl-2 3 5-tri-O-(phenylmethyl)-D-lyxofuranose (500 mg 1.15 mmol) with 4-N-[4-(dirnethyla"~i"or"eLhyl)phenyl]amino-5-phenyl-pyrrolo[2 3-d~pyrimidine (593 mg 1.5 20 mmol) provided an u~,sepa,~ble mixture of the title compound and its N1-isomer and tris[2-(2-methoxyethoxy)ethyl]amine (836 mg); Rf = 0.6 (silica gel dichloromethane/methanol 90/10).

W O 96/40705 PCT~US96/10404 Example 25 Preparalion of formula 1 - 4-N-~4-(Dimethylaminomethyl)phenyl]amino-5-phenyl-7-(4-C-methyl-~-D-lyxofuranosyl)pyrrolo~2.3-d~-pyrimidine: Table 2 # 64 An orange solution of palladium chloride (400 mg) in anhydrous ,r,e~l ,anol (10 mL) was ~le~cse~l and stirred under hydrogen (1 atm) for 10 minutes.
A solution of 4-N-[4-(dimethylaminomethyl)phenyl]amino-5-phenyl-7-(4-C-methyl-2 3 5-tri-O~phenylmethyl)-~-D-lyxofuranosyl)-pyrrolo~2 3-dlpyrimidine its N1-isomer and tris[2-(2-meU .u,cyethoxy)ethyl]amine (786 mg) in solution in anhydrous " ,e~l -a"ol (10mL) was added to the suspension of reduced palladium. The heterogeneous reaction mixture was stirred at room ter~ ,per~L~re under hydrogen (1 atm) for 6 hours filtered through Celite~ and the filtering pad was rinsed with boiling methanol. The combined filtrates were concentrated under reduced pressure. The residue was dissolved in 0.1N hydrochloric acid and washed twice with ethyl acetate. The pH of the ~ eous solution was brought to 12 with 1 N aqueous sodium hydroxide and the resulting solution was extracted 3 times with ethyl ~cePte. The combined Gryal liC
ext, ~ were dried over sodium sulfate and concentrated under red~ ~ce~i pressure.
The residue was purified by flash cl ,ro" ,aLûyl ~pl ,y on silica gel (dichloromethane/methanol/30 % aqueous ammonium hydroxide 90/10/1 to 80/2011).
The partially purified nucleoside was further purified by HPLC (C18 50X250 mm methanol/(~Ler/".ell,ar,ol/aceticacid95/5/0.5)45/55 16.5mL/minute A,~ 299nm Rt= 20.6 minutes) and cryst~lli7Pd from ethanol. Yield: 26.8 mg Rf = 0.25 (silica gel dichloromethane/methanol 80/20) m.p. 205-206~C.

Example 26 Preparation of compound of formula 12 4-C-Methyl-2 3.5-tri-O-(phenylmethyl)-1-(1.3-dithian-2-yl)-D-nbo-pentane A solution of the compound of Example 8 (4 g 10 mmol) in dry tetrahydrofuran (100 mL) was added dropwise over 10 minutes to a solution of [(phenylmethyl)oxy]methyllithium (1.8 mmol) (Still W. C. J. Am. Chem. Soc. 100, 1481 (1978)) in dry tetrahydrofuran (50 mL) at -78~ C. After stirring for 10 minutes at -78~ C the reaction mixture was quenched by slow addition of a solution of acetic acid (2.3 mL) in dry tetrahydrofuran (50 mL) over 5 minutes at -78~ C. The quenched solution was warmed to room temperature, diluted with 0thyl ~cet~te and washed with saturated ~ eous a" l, l lonium cl ,lo, ide, saturated aqueous sodium bicarbonate and saturated aqueous sodium chloride. The organic layer was dried over sodium sulfate and concenlr~Led under red~ ~ced pressure. The residue was purified by flash 5 c;hl Ul l laloyl ap hy on silica gel (hexanes/ethyl ~cet~te 90/10 to 75/25). Yield: 3.68 9, 70 %, Rf = 0.45 (silica gel, hexanes/ethyl ~cet~te 70/30).

ExamPle 27 P, eparaLion of compound of formula 2 4-C-Methyl-2,3.5-tri-0-(phenylmethyl)-D-ribofuranose The title compound was synthesi7ed following a procedure analogous 10 to the synthesis described in Example 4. Thus the compound of Example 26 (3.68 9, 7 mmol) gave 2.22 9, 73 %, Rf = 0.25 (silica gel, hexarles/ethyl aceta~e 70/30).

Example 28 Preparation of compound of formula 4 4-N-Phenylamino-5-Phenyl-7-(4-C-methyl-2,3 5-tri-O-(phenylmethyl)-~-D-ribofuranosyl)pyrrolor2~3-~pyrimidine The title compound was synthes;~d following a procedure analogous to the synthesis described in Example 12. Thus coupling of 4-C-methyl-2,3,5-tri-O-(phenylmethyl)-D-ribofuranose (500 mg,1.15 mmol) with 4-N-phenylamino-5-phenyl-pyrrolo[2,3~pyrimidine (494 mg,1.73 mmol) provided the titled nucleoside (165 mg, 20 %); Rf = 0.6 (silica gel, hexanes/ethyl acetate 70/30).

20 Example 29 Preparation of compound of formula 1 4-N rl ,en~lamino-5-phenyl-7-(4-C-methyl-B-D-ribofuranosyl)pyrrolo~2.3-dlr yrimidine:
Table 2 ~81 The title compound was synthesized following a procedure analogous to the synthesis described in Example 24. Thus 4-N-phenylamino-5-phenyl-7-(4-C-25 methyl-2,3,5-tri-0-(phenylmethyl)-~-D-ribofuranosyl)pyrrolo-~2,3-o~pyrimidine (144 mg) provided the titled deprotected nucleoside (63 mg, 73 %), Rf = 0.45 (silica gel, dichloromethane/methanol 90/10), m.p. 211 -213~ C.

W O 96/40705 PCTrUS96/10404 Example 30 Preparation of comDound of formula 4 t 4-N-~4-(Dimethyiaminomethyl)phenyl]amino-5-phenyl-7-(2.3-O-(methylethylidene)4-C-spirocyclopropyl-~-D-erythrofuranosyl)pyrrolo[2.3-dloyrimidine The title compound was synthesized following a procedure analogous to the synthesis described in Example 12. Thus coupling of 2,3-O~methylethylidene)-4-C-spirocyclopropyl-D-erythrofuranose (350 mg, 1.88 mmol) with 4-N-[4-(dimethyld" ,i"omell ,yl)phenyl]amino-5-phenyl-pyrroloE2,3-o~pyrimidine (1.1 1 9, 3.99 mmol) provided an unseparable mixture of the titled nucleoside, its N1-isomer and tris[2-(2-methoxyethoxy)ethyl~amine (1.31 9); Rf = 0.45 (silica gel, dichloromethane/methanol 90/10).

ExamDle 31 P,eparalion of comDound of formula 1 4-N-~4-rDimethylaminomethyl)phenyllamino-5-phenyl-7-(4-c-spirocvclopropyl-B-D
erythrofuranosyl)~yrrolo-[2.3-dl~yrimidine: Table 4 #158 A mixture of 4-N-[4-(dimethylaminomethyl)phenyllamino-5-phenyl-7-1 5 (2,3-0-(methylethylidene)4-C-spirocyclopropyl-~-D-erythrofuranosyl)pyrrolo[2,3-dlpyrimidine, its N1-isomer and tris[2-(2-methoxyethoxy)ethyl]amine (1.31 g) wasdissolved in methanol (10 mL) and 0.1N hydrochloric acid (10 mL). The pH was adjusted to pH=1.5 with 6N hydrochloric acid (0.5 mL) and the homogeneous solution was refluxed for one hour. The reaction mixture was diluted with 0.1 N hydrochloric acid and washed twice with ethyl acetate. The pH of the aqueous solution was brought to 12 with 1 N aqueous sodium hydroxide and the resulting solution was ext, d~ed 3 times with ethyl acetate. The combined organic extracts were dried over sodium sulfate and concer,l, aled under reduced pressure. The residue was purified by flash chromatography on silica gel (dichloromethane/methanol/30 % aqueous ammonium hydroxide 90110/1 to 80/20/1). The partially purified nucleoside was further purified by HPLC (C18, 50X250 mm, methanol/(water/methanol/acetic acid 9515/0.5) 45/55, 18 mUminute, AmaX= 299 nm, Rt= 17 minutes) and crystallized from ethyl acetate to provide the title compound (Rf = 0.25 (silica gel, dichloromethane/methanol 80/20). MS, calculated (M+H) = 472.23; found = 472.

W O 96/40705 PCT~US96/10404 Example 32 Preparation of compound of formula 4 4-N-Phenylamino-5-Phenyl-7-(2~3-0-(methylethylidene)-B-D-erythrofuranosyl)pyrrolo~2, 3-d~pyrimidine Oxalyl chloride (.55 mL, 6.3 mmol) was added dropwise, keeping the 5 Ler,l5~er~lure below35~C, to a solution of N,N-dimethylro,."ai,lide (4.8 mL, 63 mmol) in toluene (5.4 mL) and aceLoni~ ile (1.9 mL). The slushy mi~ture was stirred at room temperature for 15 minutes then cooled to -12~C. A solution of 2,3-O-(methylethylidene)-~-D-erythrofuranose (1 9, 6.24 mmol)( Cohen, N. ef al. J. Am.Chem. Soc. 105, 3661 (1983)) in toluene (1.2 mL) was added to the reaction mixture 10 maintaing the temperature below -12~ C. After stirring at -12~ C for 20 minutes the solution was cooled to -16~C and a solution of triethylarlline (1.1 mL, 7.9 mmol) in toluene (1 mL) was added maintening the temperature below 0~ C. The precipitate was stirred 15 minutes at 0~C, filtered off over a pad of Celite~ and rinsed with toluene. The combined filtrates were added to a mixture of 4-N-phenylamino-5-15 phenylpyrrolo[2,3-~pyrimidine (2.85 g, 1 mmol), finely powdered pot~ssil ~m hydroxide (85%, 1.31 g, 2 mmol) and tris[2-(2-methoxyethoxy)ethyl]amine (4 mL, 1.25 mmol) in dry toluene which had been stirring at room t~mperature for 2 hours.
After stirring overnight at room temperature, the reacLiGI, rnixture was diluted with ethyl :~ceL~la and washed with saturated aqueous a, l Imo, lium chloride. The organic 20 layer was dried over sodium sulfate and conce, lll ~Led under reduced pressure. The residue was purified by flash chromatography on silica gel (hexanes/ethyl ~cet~le 70/30 to 50/50). Yield: 969 mg, 36 %, Rf = 0.55 (silica gel, hexanes/ethyl acetate 60/40).

CA 02220642 1997-ll-28 Example 33 Preparation of compound of formula 1 4-N-Phenylamino-5-phenyl-7-(~-D-erythrofuranosyl)pyrrolo~2.3-d~pyrimidine: Table1 ~27 The title compound was synthesi~d following a procedure analogous 5 to the sy, llhesis des~ i~ed in Example 13. Thus 4-N-phenylamino-5-phenyl-7-(2,3-O-(methylethylidene)-~-D-erythrofuranosyl)pyrrolo[2,3-~pyrimidine (969 mg, 2.26 mmol) provided the title nucleoside (401 mg, 46 %), Rf = 0.5 (silica gel, dichloromethane/l ,~elhar ~ol 90/10), m.p. 210.5-211.5~ C.

ExamDle 34 P, et~ara~ion of compound of formula 12 10 4-C-~(M~;l ,oxy)methyl]-2 3.5-tri-O-(phenylmethyl)-1 -(1.3-dithian-2-yl)-D-/yxo-pentane A solution of the compound of Example 2 (1.02 9, 2 mmol) in dry tetrahydrofuran (40 mL) was added dropwise over 5 minutes to a solution of [(methyl)oxy]methyllithium (6 mmol) (Still, W. C. J. Am. Chem. Soc. 1978, 100, 1481) in dry tetrahydrofuran (40 mL) at -78~ C. After stirring for 20 minutes at -78~ C, the 15 reaction mixture was quenched with saturated ~queo~ ~s a~ ~ ~monium chloride, warmed to room temperature, diluted with ethyl acetate and washed with saturated aqueous ammonium chloride. The organic layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by flash chroma-tography on silica gel (hexanes/ethyl acetate 90/10 to 75/25). Yield: 0.48 g, 43 %, 20 Rf = 0.45 (silica gel, hexanes/ethyl acetate 70/30).

Example 35 Preparation of compound of formula 2 4-C-Methoxymethyl-2.3.5-tri-0-(phenylmethyl)-D-lyxofuranose The title compound was synthesized following a procedure analogous to the synthesis described in Example 4. Thus the compound of Example 34 (3.53 25 g,6.3 mmol) gave the title compound (1 g,34 %), Rf = 0.25 (silica gel, hexanes/ethyl acetate 70/30).

W O 96/40705 PCT~US96110404 Example 36 Preparation of compound of formula 4 4-N-(4-Fluorophenyl)amino-5-phenyl-7-(4-C-methoxymethyl-2.3,5-tri-O-(phenylmethyl)-B-D-lyxofuranosyl)pyrrolo~2.3-~lpyrimidine The title c ompound was synthesi7~d followin~1 a procedure analogous 5 to the synthesis described in Example 12. Thus coupling of 4-C-methoxymethyl-2,3,5-tri-O-(phenylmethyl)-D-ly~cofuranose (500 mg, 1.08 mmol) with 4-N-(4-fluoro,ul ,enyl)amino-5-phenylpyrrolo[2,3~p~ ll;dil ,e (0.5 9, 1.64 mmol) provided the title compound (328 mg, 40 %); Rf = 0.6 (silica gel, hexanes/ethyl acetate 70/30).

Example 37 Preparation of compound of formula 1 10 4-N-(4-Fluorophenyl)amino-5-phenyl-7-(4-C-methoxymethyl-~-D-lyxofuranosvl)Pyrrolo~2.3-dlPyrimidine: Table 3 ~372 The title compound was synthesized following a proce.l.lre analogous to the synthesis desc~ ibed in Example 25. Thus 4-N-(Fluor~phe"yl)amino-5-phenyl-7-(4-C-methoxymethyl-2,3,5-tri-O-(phenylmethyl)-~-D-lyxofuranosyl)pyrrolo-[2,3-15 ~pyrimidine (430 mg) provided the titled depr~tected nucleoside Rf = 0.5 (silica gel,dichloromethanelmethanol 90/10). MS, calu ll~ted (M+H)= 481; found = 481. m.p.
205-206 ~C.

ExamDle 38 Preparation of Compound of formula 4 1 -(2.3-Di-O-acetyl-,B-D-erythrofuranosyl)-3-(4-chlorophenyl)-4-N-20 phenylaminopyrazolo~3.4-dlpyrimidine A mixture of 1 ,2,3-tri-O-acetyl-D-erythrofuranose (619 mg, 2.5 mmol) obtained accordi"g to Kline, J.Org.Chem. 57:6, 1772 (1992), 3~4 chlc),~ l,enyl)-4-N-phenyla" ,L ,o~yrazolo[3,4-d]pyrimidine (809 mg, 2.51 mmol.) obtained accor~ lg to the methods in U.S. arplir~tion Serial No. 08/014,190, and boron trifluoride diethyl 25 etherate (620 ,uL; 5 mmol) in nitromethane (20 mL) was refluxed for 1 hour. After cooling to room temperature, the reaction mixture was conc:entrated under reduced pressure. The residue was purified by flash chromatography (silica, hexanes/ethyl ~3~1e 80/20 to 50/50) to provide the protected nucleoside (625 mg, 49%), Rf=0.2 (silica, hexaneslethyl acetate 70/30).

CA 02220642 l997-ll-28 W O 96140705 PCT~US96/10404 Example 39 Preparation of compound of formula 1 3-(4-Chlorophenyl)-1 -(B-D-erythrofuranosyl)-4-N-phenylaminopyrazolo[3.4-dlovrimidine: Table 5 #432 A 0.5 M solution of sodium methoxide in methanol (2.4 mL, 1.2mmol) 5 was added to a solution of 1-(2,3-di-O-acetyl-~-D-erythrofuranosyl)-3-(4-chlorophenyl)-4-N-phenylaminopyrazolo~3,4-d]pyrimidine (308 mg, 0.6 mmol) in methanol (10 mL) at 0 ~C. After stirring at 0 ~C for 30 minutes, the reaction mixture was quenched with acetic acid (0.25 mL) and conce"l~aled under red~ Iced pressure.
The residue was purified by ~ " um aLuyl ~phy (silica, dichloromethane/methanol 96/4 10 to 90/10). Recryst~ tion from ethanol afforded the pure product. Rf=0.6, (silica, dichloromethane/methanol 90:10), m.p.194-195~C.

Example 40 Preparation of compound of formula 4 4-N-(4-Fluorophenyl)amino-5-phenyl-7-(2,3-O-(methylethylidene)-B-D-erythrofuranosyl)pyrrolo~2.3-dlPyrimidine The title compound was synthesized following a procedure analogous to the synthesis described in Example 32. Thus coupling of 2,3-O-(methylethylidene)-~ -D-erythrofu, ~"ose (5.6 g, 35 mmol) with 4-N-(4-fluorophenyl)amino-5-phenylpyrrolo[2,3-d]pyrimidine (15.9 9, 52.5 mmol) provided the title n~ ~clcoside (4.58 9,29 %); Rf = 0.5 (silica gel, hexanes/ethyl acetate 80/20).

20 Examole 41 Preparation of compound of formula 1 4-N-(4-Fluorophenyl)amino-5-phenvl-7-(B -D-erythrofuranosyl)pyrrolo~2.3-dl~yrimidine: Table 1 #28 A solution of 4-N-(4-fluorophenyl)amino-5-phenyl-7-(2,3-O~methylethylidene)-~ -D-erythrofuranosyl)pyrrolo[2,3-d]pyrimidine (3.26 9) in 70 % aqueous 25 trifluoroacetic acid (30 mL) was stirred at rt for 1 hour. The reaction mixture was conce, ~ lecl under reduced pressure and azeotroped twice with water and twice with ethanol. The residue was purified by chromatography (silica, dichloromethane/methanol 95/5 to 90/10). Recryst~ tion from ethanol afforded the pure product (2.35 g, 79 %); Rf = 0.5 (silica, dichloromethane/methanol 90/10), m.p.
30 194-195~C.

CA 02220642 l997-ll-28 WO 96/40705 PCT~US96/10404 Example 42 Preparation of comoound of formula 4 4-N-(4-Fluorophenyl)amino-5-(4-fluorophenyl)-7-(2,3-O-(methylethylidene)-~-D-erythrofuranosyl)pyrrolo[2,3-dlPyrimidine The title compound was synthesized following a procedure analogous to the 5 synthesis desc~ il,ed in Example 32. Thus c oupling of 2,3-O-(methylethylidene)-~ -D-~ erythrofuranose (1 9, 6.2 mmol) with 4-N-(4-fluorophenyl)amino-5-(4-fluoropl ,enyl)pyrrolo[2,3-d3pyrimidine (3 g, 9.3 mmol) provided the title nucleoside (580 mg,20 %); Rf = 0.6 (silica gel, hexaneslethyl acetate 70/30).

Example 43 Preparation of compound of formula 1 1 0 4-N-r4-Fluorophenyl)amino-5-(4-fluorophenyl)-7-(~-D-erythrofuranosvl)pyrrolo~2.3-dlPyrimidine: Table 1 ~29 The title c ompound was synthesized following a procedure analogous to the synthesis described in Example 41. Thus 4-N-(4-fluorophenyl)amino-5-(4-fluorophenyl)-7-(2,3-O-(methylethylidene)-~ -D-erythrofuranosyl)pyrrolor2,3-1 ~ d~pyrimidine (580 mg) provided the title compound (244 mg, 41 %); Rf = 0.7 (silic a gel, dichloromethanelmethanol 80120), m.p. 200-202~C.

Example 44 Preparation of compound of formula 4 4-N-(4-Chlorophenyl)amino-5-phenyl-7-(2 3-O-(methylethylidene)-B-D-erythrofuranosyl)pyrrolo~2,3-dlPyrimidine The title c ompound was synthesized following a proc:edure analogous to the synthesis described in Example 32. Thus coupling of 2,3-O-(methylethylidene)-,~ -D-erythrofuranose (900 mg, 5.6 mmol) with 4-N-(4-chlorophenyl)amino-5-phenylpyrrolo[2,3-d]pyrimidine (900 mg,2.8 mmol) provided lhe title nucleoside (744 mg, 57 %); Rf = 0.7 (silica gel, hexaneslethyl acetate 70130).

CA 02220642 1997-ll-28 W O 96/40705 PCT~US96/10404 4~

Example 45 Preparation of compound of formula 1 4-N-(4-Chlorophenyl)amino-5-phenyl-7-(i3 -D-erythrofuranosyl)pyrrolo~2.3-dloyrimidine: Table 1 ~299 The title compound was synthesized following a procedure analogous to the 5 sy"li ~esis desc ribed in Example 41. Thus 4-N~4-chloro, l ~el ,yl)amino-5-phenyl-7-(2,3-O-(methyiethyiidenei-i3 -D-ery~hrorTurarlosyl)pyrrolo[2~3-d]pyrimidin~ (744 m, ~) provided the title compound (500 mg, 74 %); Rf = 0.5 (silica gel, dichloromethane/methanol 90/10), m.p. 212-213~C.

Example 46 Preparation of compound of formula 4 1 0 4-Chloro-5-iodo-7-(2.3-0-(methYlethylidene)-B-D-erythrofuranosyl)pyrrolor2.3-d]Pyrimidine The title compound was synthesized following a procedure analogous to thesynthesis described in Example 32. Thus coupling of 2,3-O-(methylethylidene)-~ -D-erythrofuranose (3.8 9, 21.8 mmol) with 4-chloro-5-iodopyrrolo~2,3-d]pyrimidine 15 (Pudlo, J. S. J. Med. Cem. 1990, 33, 1984, 2.54 9, 10.9 mmoij provided ihe iil;Q
nucleoside (2.64 9, 62 %); Rf = 0.7 (siiica gel, hexanes/ethyl acetate 70/30).

Example 47 Preparation of comDound of formula 1 4-Amino-5-iodo-7-(-B-D-erythrofuranosyl)pyrrolo~2 3-dlpyrimidine: Table 1 ;5~300The title compound was synthesized following a procedure analogous to the 20 synthesis described in Example 21. Thus 4-chloro-5-iodo-7-(2,3-O-(methylethylidene)-3 -D-erythrofuranosyl)pyrrolo[2,3-d]pyrimidine (714 mg) gave after chromatography (silica gel, dichloromethane/methanol 95/5 to 85/15) and crystallization from ethanol the title nucleoside (270 mg, 40 %), m.p. 258~ C (dec).

PCT~US96/10404 Example 48 r~ ~parcLiGl, of con ,pound of formula 4 4-N-~4-(2~(1.1 -Dimethylethyl)dimethylsilyloxv)ethyl)phenyl~amino-5-phenyl-7-(2.3-~(methylethylidene~-B -D-erythrofuranosyl)pyrrolo~2.3-dlPyrimidine The title compound was synthesized following a procedure analogous to the 5 synthesis des~iL,ed in Example 32. Thus coupling of 2,3-O-(methylethylidene)-~ -D-erythrofuranose (1 9. 6.2 mmol) with 4-N-[4-(2-((1,1 -dimethylethyl)~li" ,~lh~/lsilyloxy)ethyl)phenyl]amino-5-phenylpyrrolo[2,3-d]pyrimidine (1 9, 2.2 mmol) provided the title nucleoside (551 mg, 4.7 %); Rf = 0.7 (silica gel, hexanes/ethyl acetate 70/30).

10 Example 49 Preparation of comPound of formula 4 4-N-f4-(2-hydroxyethyl)Phenyl~amino-5-phenyl-7-(2.3-O-(methylethvlidene)-~-D-ervthrofuranosyl)pvrrolo~2.3-dlPyrimidine A mixture of tetraethylammonium fluoride hydrate (0.86 9, 5.76 mmol) and 4-N-~4-(2-((1,1 -dimethylethyl)dimethylsilyloxy)ethyl)phenyllamino-5-phenyl-7-(2,3-O-15 (methylethylidene)-~ -D-erythrofuranosyl)pyrrolo[2,3-d]pyrimidine (2.7 9, 4.6 mmol) in dimeth~,lru~ ,, ,ide (55 mL) was stirred at rt for 18 hours. The reaction mixture was concentrated under reduced pressure, diluted in ethyl acetate and washed with saturated ~q~ ~eo~ ~c a" " "onium chloride. The aqueous layer was back extracted with ethyl ~r~et~te and the combined organic extracts were dried over sodium sulfate and 20 concentrated under redl ~ced pressure. The residue was purified by chromatography (silica gel, hexaneslethyl acelaLe 90110 to 70130) to give the title compound (2.12 9, 98 ~/0); Rf = 0.2 (silica gel, hexanes/ethyl acetate 70130).

ExamPle 50 Preparation of compound of formula 4 4-N-~4-(2-(4-MG"~,holi, ~o)ethyl)phenyl]amino-5-Phenyl-7-(2~3-o-(methylethylidene)-l3 25 -D-erythrofuranosyl)~yrrolo~2.3-dlr~yrimidine A mixture of 4-N-[4-(2-hydroxyethyl)phenyl]arnino-5-phenyl-7-(2,3-0-(methylethylidene)-,~ -D-erythrofuranosyl)pyrrolo[2,3-d~pyrimidine (158 mg, 3.34mmol) and methyl triphenoxyphosphonium iodide (460 mg, 1 mmol) in dichloromethane (6 mL) was stirred overnight at rt. The reaction mixture was 30 quenched with methanol (1 mL) and poured into a 0.5 M solution of sodium W O 96/40705 PCTrUS96/10404 thiosulfate and extracted with ethyl acetate. The combined organic extracts werewashed with water and saturated aqueous sodium chloride, dried over sodium sulfate and concenL~ ~led under red~ ~ced pressure. The residue was dissolved in dioxane (30 mL) and morpholine was added (0.84 mL, 9.6 mmol). The resulting solution was 5 refluxed for 24 hours. After cooling to rt, the reaction mixture was diluted with ethyl le and washed with saturated aq~ ~eol ~-s ammonium chloride, dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, dichloromethane/methanol 95/5) to give the title compound, Rf = 0.7 (silica gel, dichloromethane/methanol 90/10).

10 Example 51 Preparation of compound of formula 1 4-N-~4-(2-(4-Morpholino)ethyl)phenyllamino-5-phenyl-7-(B-D-erythrofuranosyl)Dyrrolo~2.3-dlPyrimidine: Table 1 #301 The title compound was synthesized following a procedure analogous to the synthesis described in Example 41. Thus 4-N-[4-(2-(4-15 morpholino)ethyl)phenyl]amino-5-phenyl-7-(2,3-O-(methylethylidene)-~ -D-erythrofuranosyl)pyrrolo~2,3-d]pyrimidine (1.7 g) provided the title coi"pound (150 mg, 10 %) after recrys~lli7~tion from methanol/water; Rf = 0.1 (silica gel, dichloromethane/methanol 90/10), m.p.128-130~ C.

Exam~le 52 Preparation of compound of formula 4 20 4-N-~4-(2-(1-(4-feff-Butyloxyca, L u"ylpiperazino))ethvl)phenyl]amino-5-phenyl-7-(2.3-O-(methylethylidene)-~ -D-erythrofuranosyl)pyrrolo~2.3-dl~yrimidine The title compound was synthesized following a procedure analogous to the synthesis described in Example 50. Thus 4-N-[4-(2-hydroxyethyl)phenyl]amino-5-phenyl-7-(2,3-0-(methylethylidene)-~ -D-erythrofuranosyl)pyrrolo[2,3-d]pyrimidine 25 (701 mg, 1.48 mmol) and substituting feff-butyl 1-piperazinecarboxylate (830 mg, 4.46 mmol) for morpholine provided the title compound; Rf = 0.45 (C18, methanol/0.1 N hydrochloric acid 50/50).

Example 53 Preparation of compound of formula 1 4-N-[4-(2-(1 -Piperazino)ethyl)phenyl]amino-5-phenyl-7-(B-D-erythrofuranosyl)pyrrolo~2.3-d1cyrimidine trihydrochloride salt: Table 1 3~43 A s o I u t i o n o f 4 - N - [ 4 - ( 2 - ( 1 - ( 4 - f e rt-5 butyloxycarbonylpiperazino))ethyl)phenyl]amino-5-phenyl-7-(2,3-O-(methylethylidene)-~-D-erythrofuranosyl)pyrrolo[2,3-d]pyrimidine (931 mg) in 70 %
~1 leol IC trifluoroacelic acid (65 mL) was stirred at OC for 1 hour and at rt for 1 hour.
The reaction mixture was concel,l,aled under redl~ced pressure and azeotroped twice with water and twice with ethanol. The residue was purified by HPLC (C18, 10 50X250 mm, methanol/(0.1 % aqueous trifluoroacetic acid 50/50, 15 mL/minute, A,~"= 260 nm, Rt= 15.7 minutes) and Iyophilized to give the title compound (339 mg, 37 %); Rf = 0.75 (C18, methanol/0.1 N hydrochloric acid 50/50), m.p. 150-180~C
(dec).

Example 54 P, epar~Lion of compound of formula 9 15 Methyl 5-O-methyl-2.3-di-O-(phenylmethyl)-D-ribofuranoside A solution of methyl 5-~methyl-D~ibofuranoside (Dubois L. et al. Tefrahedron 1993, 49(4), 901-910, 2 9, 11.2 mmol) in dimeth~lru""a"lide (10 mL) was added dropwise to a suspension of sodium hydride (60 % in oil, 2.25 9, 563 mmol) in dimethylformamide (54 mL). After stirring at rt for 45 minutes, a solution of benzyl 20 bromide (4 mL, 33.6 mmol) was added dropwise in dimethylformamide (4 mL). Thereaction mixture was stirred at rt overnight, quenched with methanol and concentrated under reduced pressure. The residue was dissolved in ethyl acetate and washed with saturated ~ eo~ls sodium chloride. The organic layer was dried over sodium sulfate and concer,LIaled under reduced pressure. The residue was 25 purified by flash cl ,ror"~lography ( silica, hexaneslethyl acetale 70/30 to 50150) to provide the a anomer (2.86 9, 71 %) and the ,B anomer (0.86 9, 21 %); a anomer Rf = 0.5, ,~ anomer Rf = 0.4 (silica, hexaneslethyl acetate 50150).

Example 55 Pl eparalion of compound of formula 10 5-0-Methyl-2.3-di-0-(Phenylmethyl)-1-(1 3-dithian-2-yl)-D-nbo-pentane The title compound was synthesized following a procedure analogous to the synthesis described in example 1. Thus methyl 5-0-methyl-2,3-di-O-(phenylmethyl)-5 D-ribofuranoside (3.72 g, 10.4 mmol) gave the title compound (3.74 9, 83 %), Rf =
0.45 (silica, hexaneslethyl acetate 50/50).

Example 56 Preparation of compound of formula 11 (3S. 4R)-1-Methoxy-3 4-di-[(phenylmethyl)oxv]-5-(1 3-dithian-2-yl)pentan-2-one The title compound was synthesized following a procedure analogous to the 10 synthesis described in example 2. Thus 5-0-methyl-2,3-di-O-(phenylmethyl)-1 -(1,3-dithian-2-yl)-D-r7b~pentane (3.74 g, 8.61 mmol) gave the titie compound (3.32 g, 89 %), Rf = 0.3 (silica, hexaneslethyl acetate 70130).

Example 57 Preparation of compound of formula 12 4-C-Methoxymethyl-2 3.5-tri-O-(phenylmethyl)-1-(1.3-dithian-2-yl)-D-nbo-pentane The title compound was synthesized following a procedure analogous to the synthesis described in example 3. Thus (3S, 4R)-1-methoxy-3,4-di-[(phenylmethyl)oxy]-5-(1,3-dithian-2-yl)pentan-2-one (3.32 9, 7.67 mmol) gave the title compound (1.47 9, 34 %); Rf = 0.45 (silica, hexaneslethyl acetate 70130).

Example 58 Preparation of compound of formula 2 20 4-C-Methoxymethyl-2.3.5-tri-O-(Phenvlmethyl)-D-ribofuranose The title compound was synthesized following a procedure analogous to the synthesis described in example 4. Thus 4-C-methoxymethyl-2,3,5-tri-0-(phenylmethyl)-1-(1,3-dithian-2-yl)-D-nb~pentane (1.47 9, 2.65 mmol) gave the title compound (0.91 9, 74 %); Rf = 0.2 (silica, hexaneslethyl acetate 70130).

W096/40705 PCT~US96/10404 Example 59 Preparation of compound of formula 4 4-N-(4-Fluorophenyl~amino-5-~henvl-7-(4-C-methoxymethyl-2,3.5-tri-O-(phenylmethyl)-B -D-ribofuranosyl)pyrrolo~2.3-d~pyrimidine The title compound was synthesized following a procedure analogous to the 5 synthesis desc; ibed in example 12. Thus coupling of 4-C-methoxymethyl-2,3,5-tri-0-(phenylmethyl)-D-ribofi~ ose (500 mg,1.08 mmol) with 4-~V-(fluorophenyl)amino-5-phenyl-pyrrolo[2,3~pyrimidine (0.5 g,1.64 mmol) provided the title n~ ~cleoside (328 mg, 40 %); Rf = 0.5 (silica, hexanes/ethyl acetate 70/30).

ExamDle 60 Preparation of compound of formula 1 10 4-N-(4-Fluorophenyl)amino-5-phenvl-7-(4-C-methoxymethyl-B-D-ribofuranosyl)Pyrrolo[2.3-dlPyrimidine: Table 3 #352 The title compound was synthesized following a procedure analogous to the synthesis described in example 25. Thus 4-N-(4-fluoropl ,enyl)amino-5-phenyl-7-(~C-methoxymethyl-2,3,5-tri-~(phenylmethyl)-~ -D-ribofuranosyl)pyrrolo[2,3-d]pyrimidine 15 (0.69 g, 0.92 mmol)) provided the deprotected nucleoside (134 mg, 30 %), Rf = 0.4 (silica, dichloromethane/methanol 90/10), m.p.198-199~ C.

Example 61 Preparation of compound of formula 9 Methyi 5-azido-5-deoxy-D-ribofuranoside A solution of methyl 5-azido-5-deoxy-2,3-0-(methylethylidene)-D-20 ribof~ c"oside (Browne et al., U.S. Patent Serial No. 08/812,916, 15.3 g, 70.3 mmol)and para-toluenesulfonic acid monohydrate (0.69 g, 3.6 mrrol) in methanol (750 mL) was refluxed for 18 hours. The reaction mixture was quenched with pyridine (8.4 mL, 10 mmol), concentrated under reduced pressure and purified by flash chromatography (silica, hexanes/ethyl acetate 50/50 to 30/70) to provide the title 25 compound (8.19 g, 62 %); Rf = 0.15 (silica, hexanes/ethyl acetate 70/30).

W O 96/4070S PCT~US96/10404 Example 62 P- epal ~Lion of compound of formula 9 Methyl 5-azido-5-deoxy-2.3-di-O-(phenYlmethyl)-D-ribofuranoside The title compound was synthesi7~d following a procedure analogous to the synthesis descl iL,ed in example 54. Thus methyl 5-azido-5-deoxy-D-ribofuranoside 5 (B.19 9, 43.3 mmol) gave the title compound as a mixture of a anomer (2.2 9, 14 %) and the ,~ anomer (12.46 9, 78 %); ~ anomer Rf = 0.6, a anomer Rf = 0.4 (silica,hexanes/ethyl acetate 70/30).

Example 63 P,epar~lion of compound of formula 10 5-Azido-5-deoxy-2.3-di-O-(phenylmethyl)-1 -(1 .3-dithian-2-yl)-D-nbo-pentane The title compound was synthesized following a procedure analogous to the synthesis described in example 1. Thus methyl 5-azido-5-deoxy-2,3-di-0-(phenylmethyl)-D-ribofu,~,loside (12.45 9, 33.7 mmol) gave the title compound (13.48 9, 90 %), Rf = 0.45 (silica, hexanes/ethyl acetate 70/30).

Example 64 Preparation of compound of formula 11 15 (3S. 4R~-1-Azido-3.4-di-~(Phenylmethyl)oxy~-5-(1.3-dithian-2-yl)pentan-2-one The title compound was synthesized following a procedure analogous to the synthesis described in example 2. Thus 5-Azido-5-deoxy-2,3-di-O-(phenylmethyl)-1-(1,3-dithian-2-yl)-D-ribo-pentane (13.48 9, 31 mmol) gave the title compound (9.91 9, 74 %), Rf = 0.5 (silica, hexanes/ethyl acetate 70/30).

20 Example65 Preparation of compound offormula 12 4-C-Azidomethyl-5-0-[(4-methoxyphenyl)methyl~-2.3-di-O-(~henylmethyl)-1 -(1 .3-dithian-2-yl)-D-ribo-pentane The title compound was synthesized following a procedure analogous to the synthesis described in example 3. Thus reaction of [((4-25 methoxyphenyl)methyl)oxy]methyllithium (prepared using a procedure analogous tothe one described by Still, W. C. J. Am. Cem. Soc. 1978, 100,1481 for the W096/40705 PCTrUS96/10404 pr~ ion of [(phenylmethyl)oxy]methyllithium, 7.78 9, 1 ~.2 mmol) and (3S, 4R)-1 azido-3,4-di-~(phenylmethyl)oxy]-5-(1,3-dithian-2-yl)pentan-2-one (4.02 9, 9.06 mmol) gave the title compound (2.07 g, 39 %); Rf = 0.45 (silica, hexanes/ethyl ~cet~te70/30)-5 ExamPle 66 Preparation of compound of formula 2 4-C-Azidomethyl-5-0-r(4-methoxyPhenyl)methyl]-2,3-di-0-(phenylmethyl)-D-ribofuranose The title compound was synthesized following a procedure analogous to the synthesis described in example 4. Thus 4-C-azidomethyl-5-0-~(4 10 methoxyphenyl)methyl]-2,3,-di-0-(phenylmethyl)-1 -(1,3-ditllian-2-yl)-D-nbo-pentane (2.07 9, 3.47 mmol) gave the title compound (1.19 g, 68 %); Rf = 0.4 (silica, hexanes/ethyl Acet~le 70/30).

Example 67 P,~paralion of compound of formula 4 4-N-(4-Fluorophenyl)amino-5-phenvl-7-(4-C-azidomethyl-5-0-~(4-15 methoxyphenyl)methyl]-2 3-di-0-(phenylmethyl)-B-D-ribofuranosyl)pyrrolo~2.3-dlDyrimidine The title compound was synthesized following a procedure analogous to the synthesis described in example 12. Thus coupling of 4-C-azidomethyl-5-0-[(4-methoxyphenyl)methyl]-2,3~i-0-(phenylmethyl)-D-ribofuranose (1.19 mg, 2.3 mmol) 20 with 4-N-(fluorophenyl)amino-5-phenylpyrrolo[2,3-c~pyrirr~idine (2.17 9, 7.1 mmol) provided the title nuGleoside (656 mg, 35 %); Rf = 0.6 (silic:a, hexanes/ethyl ~ce~te 70/30).

Example 68 Preparation of compound of formula 4 4-N-(4-Fluorophenyl)amino-5-phenyl-7-(4-C-aminomethyl-5-0-~(4-25 methoxyphenyl)methyl~-2,3-di-0-(phenvlmethyl)-B-D-ribofuranosyl)pyrrolo[2 3-dlPyrimidine A mixture 4-N-(4-fluorophenyl)amino-5-phenyl-7-(4-C-azidomethyl-5-0-[(4-methoxyphenyl)methyl]-2,3-di-0-(phenylmethyl)-B-D-ribofuranosyl)pyrrolo[2,3-d]pyrimidine (448 mg, 0.57 mmol) and triphenylphosphine (300 mg, 0.11 mmol) in W O 96/40705 PCT~US96/10404 toluene (30 mL) was refluxed for 2 hours. The reaction mixture was quenched withlh~nol and reflux was carried on for 30 minutes. After cooling, the solution wascol lcei ,laled under red~ ~ced pressure and purified by flash chromaLoyl d~Jh y (silica, dichlor~meU,ane/lllell,allol 95/5) to provide the title compound (284 mg, 65 %), Rf =
S 0.4 (silica, dichloromethane/methanol 90/10).

Example 69 Preparation of compound of formula 1 4-N-(4-Fluorophenyl)amino-5-phenyl-7-(4-C-aminomethyl-~-D-ribofuranosyl)pyrrolo[2.3-dlPvrimidine dihydrochloride: Table 3 #392 lodc,LI i",eU~ylsilane (0.4 mL,28 mmol) was added dropwise to a solution of 4-10 N-(4-fluorophenyl)amino-5-phenyl-7-(4-C-aminomethyl-5-0-[(4-methoxyphenyl)methyl]-2,3-di-O-(phenylmethyl)-~ -D-ribofuranosyl)pyrrolo[2,3-d]pyrimidine (166 mg, 0.22 mmol) in chloroform (10 mL) at 0~C. After stirring at 0~C
for 40 minutes and rt for 24 hours, the reaction mixture was quenched with methanol and concentrated under reducerl pressure. The residue was purified by flash 15 cl,ro",alography (silica, dichloromethane/methanol/28% aqueous ammonium hydl oxide 80/20/1). HPLC (C18, 50X250 mm, l "~ll ,c, lol/0.1 % ~ql ~eous trifluoroacetic acid 65/35,15 mUminute, Ama,~= 260 nm, Rt= 21.64 minutes) and Iyophilization with 1 N h~dl u~l ,lo, ic acid gave the title compound (64 mg, 32 %); m.p.200-220 ~ C (dec).

ExamPle 70 Prepar~lion of compound of formula 4 20 4-N-(4-Fluorophenyl)amino-5-phenyl-7-(4-C-azidomethyl-2.3-di-O-(phenylmethyl)-~
-D-ribofuranosyl)pyrrolo~2,3-dlPyrimidine 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (122 mg, 0.53 mmol) was added to a solution of 4-N-(4-fluorophenyl)amino-5-phenyl-7-(4-C-azidomethyl-5-0-[(4-methoxyphenyl)methyl]-2,3-di-O-(phenylmethyl)-~-D-ribofuranosyl)pyrrolo[2,3-25 d]pyrimidine (212 mg, 0.27 mmol) in dichloromethane/water (2/1, 7.5 mL). Afterstirring at rt o~ I ,l, the reaction mixture was diluted with ethyl z~r~ l~le and washed with saturated aqueous sodium bical bo"ate and saturated aqueous sodium chloride.

W O 96/40705 PCTrUS96/10404 The organic layer was dried over sodium sulfate and conce, IL. aled under reduced pressure. The residue was purified by flash chromatography (silica, hexanes/ethyl acetate 70/30) to give the title compound (51 mg, 28 %); Rf = 0.35 (silica, hexanes/ethyl acetate 70/30).

5 Example 71 Preparation of comPound of formula 4 4-N-(4-FluoroPhenyl)amino-5-phenvl-7-(4-C-azidomethyl-5-0-[(4-methylphenyl)sulfonyl~-2.3-di-0-(phenylmethyl)-~-D-rib,ofuranosyl)pyrrolo~2,3-d1Pyrimidine para-Toluenesulfonyl chloride (76 mg, 0.4 mmol) was added to a solution of 10 4-N~4-fluorupl ,enyl)amino-~-phenyl-7-(4-C-~ido" ,ethyl-2,3-di-O-(phenylmethyl)-~-D-ribofuranosyl)pyrrolo[2,3-d]pyrimidine (51 mg, 0.08 mmol~, pyridine (0.062 mL, 0.8 mmol) and 4-dimethyla",ir,opyridine (10 mg, 0.08 mmol) in dichloromethane at 0~ C.
After stirring at 0~ C for 30 minutes, at rt for 3 days and being refluxed for 2 hours, more para-toluenesulfonyl chloride, pyridine and 4-dimethylaminopyridine was added 15 and the reaction mixture was stirred at rt for 2 days. Concenlldlion under re~ ced pressure and purification by flash chl ~,r"~Lography (silica, hexanes/ethyl :~cet~te 95/5 to 80/20) gave the title compound (51 mg, 50 %); Rf = 0.5 (silica, hexanes/ethylacetate 70/30).

Example 72 Prepar~Lion of compound of formula 4 20 4-N-(4-Fluorophenyl)amino-5-phenyl-7-(4-C-spiro('3-azetidino)-2 3-di-O-(phenylmethyl)-B -D-erythrofuranosyl)pyrrolo~2 3-dlPYrimidine A mixture of 4-N-(4-fluorophenyl)amino-5-phenyl-7-(4-C-azidomethyl-5-0-[(4-methylphenyl)sulfonyl]-2,3-di-O-(phenylmethyl)-,B-D-ribofuranosyl)pyrrolo[2,3-d]pyrimidine (31 mg, 0.038 mmol) and triphenylphosphine was refluxed for 1525 minutes, cooled to rt and concentrated under reduced pressure. The residue was purified by flash chr~r"dLography (silica, dichloromethane/methanol) to give the title pound (22 mg,94 %); Rf = 0.6 (silica, dichloromethane/methanol/1 % aqueous ammonium hydroxide 80/20/1).

W O 96/~0705 PCTAUS96/10404 Example 73 P, t:par~Lion of compound of formula 1 4-N-(4-Fluorophenvl)amino-5-phenyl-7-(4-C-sPiro(3-azetidino)-~-D-ervthrofuranosyl)pyrrolo~2.3-d~Pyrirriidine: Table 4a ;~414 The title compound was synthesized following a procedure analogous to the 5 synthesis described in example 69. Thus 4-N-(4-Fluorophenyl)amino-5-phenyl-7-(4 C-spiro(3-azetidino)-2,3-di-0-(phenylmethyl)-~-D-erythrofuranosyl)pyrrolo~2,3-d]pyrimidine (4.8 mg) gave the title compound; Rf=0.15 (silica, dichloromethane/methanol 80/20); MS, calcui~ted (M+H) = 448.16; found = 448.

Example 74 Preparation of compound of formula 4 10 ~N-~(1.1.2-Trimethylpropyl)dimethylsilyloxvmethyllohenylamino-5-phenyl-7-(2.3-O-(methylethylidene)-~-D-erYthrofuranosyl)pvrrolo~2,3-d]pyrimidine The title compound was synthesized following a procedure analogous to the synthesis described in Example 32. Thus coupling of 2,3-0-(methylethylidene)-~-D-erythrofuranose (3.6 g, 22.6 mmol) with 4-N-[(1,1,2-i, i m eLi ,yipropyi)di" ,eti ,ylsilyloxymethyl]phenylamino-5-phenylpyrrolo[2,3-d]pyrimidine (5 9, 11.3 mmol) provided the title nucleoside (4.21 9, 64 %); Rf = 0.65 (silica, hexanes/ethyl acetate 70130).

Example 75 Preparation of compound of formula 4 4-N-(Hydroxymethyl)phenylamino-5-,chenyl-7-(2 3-O-(methylethylidene)-~-D-20 erYthrofuranosyl)pyrrolo~2.3-d~Pvrimidine Triethylammonium fluoride hydrate (36 mg, 0.24 mmol) was added to a solution of ~N-[(1,1,2-~l i" leLI "/Ipropyl)di" ,e~l ,ylsilyloxymethyl]phenylamino-5-phenyl-7-(2,3-~methylethylidene)-~-D-erythrofuranosyl)pyrrolo[2,3-dlpyrimidine (101 mg,0.17 mmol) in dimethylru,,,,~l,,ide (2 mL) at rt. After stirring at rt for 30 minutes, the 25 reaction mixture was concentrated under reduced pressure. The residue was dissolved in ethyl ~ and washed with saturated aqueous sodium chloride. The organic layer was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by flash chromatography (silica, hexanes/ethyl ~ce~le 70/30 to 50/50) to provide the title compound (68 mg, 88 %); Rf = 0.2 (silica, 30 hexanes/ethyl ~cetsJte 70/30).

W O 96/4Q705 PCT~US96/10404 Example 76 P,~aralion of compound of formula 4 4-N-(Diethylaminomethyl)phenylamino-5-phenvl-7-(2 3-O-(methylethylidene)-B-D-erythrofuranosyl)pyrrolo[2.3-dlpyrimidine Triphenoxyphosphonium iodide (1 9, 2.2 mmol) was added to a solution of 5 4-N-(hydroxymethyl)phenylamino-5-phenyl-7-(2,3-O-(rnethylethylidene)-~-D-erythrofuranosyl)pyrrolo[2,3-d]pyrimidine (500 mg,1.09 rnmol) in dichloromethane(4 mL) at rt. After stirring at rt for 30 minutes, diethylamine (0.46 mL, 4.5 mmol) was added and stirring was carried on at rt overnight. The reaction mixture was thendiluted with ethyl acel~le and washed with 0.5 N aqueous sodium thiosulfate, 10 saturated ~ql~eous sodium bicarbonate, water and saturated aq~so~s sodium chloride. The organic layer was dried over sodium sulfate and concentrated underreduced pressure. The residue was purified by flash chromatography (silica, hexanes/ethyl ~c~l~le 75/25 to 25/75) to provide the title compound (469 mg, 84 ~/0);
Rf = 0.1 (silica, hexanes/ethyl acetate 70/30).
15 Example 77 Preparation of compound of formula 1 4-N-(Diethyla"~;, lcs" ,~ll ,yl)phenylamino-5-phenyl-7-(B-D-ervthrofuranosyl)pyrrolo[2.3-d10yrimidine dihydrochloride salt: Table 1 ~303 ~ N-(Diethyl~" ,i"o" ,elhyl)phenylamino-5-phenyl-7-(2,3-O-(methylethylidene)-B-D-erythrofuranosyl)pyrrolo[2,3-d]pyrimidine (2 mmol) was dissolved in 70%
20 aqueous trifluoroacetic acid and stirred at rt for 2 hours. The volatiles were evaporated and the residue was coevaporated with water (2X20 mL) and ethanol (2X20 mL). The residue was purified by HPL(, (C18, 50X250 mm, ",ell,~..ol/(water/0.1% trifluoroacetic acid) 50/50), 15 mL/rninute, A,r,a,~- 260 nm, Rt=
23.4 minutes) and Iyophilized three times with 0.5 N hydrochloric acid to afford the 25 pure product Rf = 0.3 (silica, dichloromethane/methanol/~8% aqueous ammonium hydroxide 80/20/1); m.p. 80-140~C (dec).

CA 02220642 l997-ll-28 WO 96140705 PCT~US96/10404 Example 78 Preparation of compound of formula 4 4-N-~1 -(4-feff-Butyloxycarbonylpiperazino)methyllDhenYlamino-5-phenyl-7-(2~3 (methylethylidene)-B-D-erythrofuranosyl)pvrrolo~2.3-dl~yl ir~idine The title compound was synthesized following a procedure analogous to the synthesis described in Example 76. Thus 4-N-(hydroxymethyl)phenylamino-5-phenyl-7~2,3-O-(methylethylidene)-~-D-erythrofuranosyl)pyrrolo[2,3-d]pyrimidine (100 mg, 0.22 mmol) and feff-butyl 1-piper~inecarboxylate (160 mg, 0.86 mmol) gave the title compound (109 mg, 79 %); Rf = 0.35 (silica, hexanes/ethyl ~cet~te 50150).

Example 79 PreParation of comPound of formula 1 4N-(1-Pi~ er~i"omethyl)phenylamino-5-Dhenyl-7-(B-D-erythrofuranosyl)pyrrolo~2.3-dlPyrimidine dihydrochloride salt: Table 1 ;~304 The title compound was synthesized following a procedure analogous to the synthesis described in Example 77. Thus 4-N-[1-(4-teff-butyloxycarbonylpiperazino)methyl~phenylamino-5-phenyl-7-(2,3-0-(methylethylidene)-~-D-erythrofuranosyl)pyrrolo~2,3-d]pyrimidine gave the title compound; MS, calculated (M~H~ = 486.58; found = 487.

Example 80 Preparation of compound of formula 4 4-N-(2-N-Phthalimidoethyl)phenylamino-5-phenyl-7-(2,3-O-(methylethylidene)-~-D-erythrofuranosyl)pyrrolo~2.3-d1Pyrimidine Diisopropyl azodicarboxylate (0.75 mL, 3.75 mmol) was added to a clear solution of 4-N-(2-hydroxyethyl)phenylamino-5-phenyl-7-(2,3-~(methylethylidene)-~-D-erythrofuranosyl)pyrrolo[2,3-d]pyrimidirie (1.2 9, 2.5 mmol), triphenylphosphine (1 g,3.75 mmol) and phthalimide (560 mg,3.75 mmol) in tetrahydrofuran (25 mL) at rt.
After stirring at rt for 3 hours, the reaction mixture was concentrated under redl Iced pressure and the residue was purified by column chromatography (silica, - hexanes/ethyl acetaLe 70/30 to 50/50) to give the title compound; Rf = 0.55 (silica, hexanes/ethyl acetate 50/50).

CA 02220642 l997-ll-28 W O 96/40705 PCT~US96/10404 Example 81 Pre,.)ar~Lion of compound of formula 4 4-N-(2-aminoethyl)phenylamino-5-phenyl-7-(B-D-erythrofuranosyl)pyrrolo~2.3-dlPyrimidine A mixture of 4-N-(2-N-phthalimidoethyl)phenylamino-5-phenyl-7-(2,3-O-5 (methylethylidene)-~-D-erythrofuranosyl)pyrrolo[2,3-d]pyrimidine (2.76 9, 4.5 mmol) and 97% hydrazine (0.8 mL) in ethanol (25 mL) was r~ fluxed for 2 hours. After cooling to rt, the white precipitate that formed during the reaction was filtered off and rinsed with ethanol. The combined filtrates were concentrated under red~cecl pressure and the residue was dissolved in 70% aqueou; trifluoroaceLic acid. After 10 stirring at rt for 2 hours, the reaction mixture was concentrated under red~ced pressure and azeotroped twice with water and twice with ethanol. The residue waspurified by column chror,,~Luyl~p hy (silica, dichloromethane/methanol/28% aqueous ammonium hydroxide 90/10/1 to 70130/1) to provide the title compound; Rf = 0.3 (silica, dichloromethane/methanol 80/20).

15 Example 82 Preparation of compound of formula 1 4-N-(2-auanidinoethyl)phenylamino-5-phenyl-7-(B-D-ery~hrofuranosyl)pyrrolo~2.3-dlPyrimidine: Table 1 i~309 The title compound is made following the procedure of M. S. Bernatowicz ef al. J. Org. Chem. 57, 2497 (1992).

20 Example 83 Preparation of the Representative Heterocycles Heterocycles, as shown in Scheme 14, are made in the following manner.

A. Preparation of compound of formula 37 (2-Amino-3-cyano4-phenylpyrrole) To a solution of phenacyl chloride (500 9, 3.23 M) in dry N,N-25 dimethylformamide (600 mL) was added potassium phthalimide, (600 9, 3.23 M) in small portions. The resulting mixture was stirred at ambient temperature overnight.
To this was added malononitrile (256 9, 3.88 M) in one lot followed by a 25 wt %solution of sodium methoxide in methanol (744 mL, 3.2 mol). The resulting mixture CA 02220642 1997-ll-28 W O 96/40705 PCT~US96/10404 was stirred at room tempera~ure overnight. Ice-water (10.0 L) was added to the reaction mix~ure and stirring was continued at room temperature overnight. The precipitate fommed was collected by rilll~Lion and washed with cold water (4.0 L). The off-white solid was stirred in toluene (3.0 L) and filtered. The solid was washed with 5 toluene (300 mL) and dried under vacuum at 60 C overnight. Yield 298.56 g. m.p.
172-174~C.

B. Preparation of compound of formula 39 (5-Phenyl4-N-(4-fluorophenyl)aminopyrrolo~2.3-dlPyrimidine) A mixture of the compound of Examlple 39A (296.0 9, 1.62 mol) and triethylol U ,Oro" ,~Le (3.2 L) was reflu~ced for 1 h. The triethylo, Ll ,ur." " ,~le was distilled off under reduced pressure un~il the pot temperature reached 88~ C. To the cooled rt:a~;~;o" mixture hexane(3.0 L) was added with vigorous stirring. The contents of the vessel were cooled to 0~ C and the off-white solid formed was collected by filtration 15 and washed with hexane (2x~00 mL) and dried under suction. Final drying was done in a high vacuum oven. Yield of the 2-ethoxymethylene-3-cyano~phenylpyrrole was 323.0 9 (83%). m.p. 98-100~ C.
The above material (100 g, 0.42 mol) was dissolved in 1,2-dichlorobenzene. 4-Fluoroaniline(60 mL, 0.62 mol) was added and the reaction 20 mixture was heated to 125~C for 1 h. An addiLional 985 mL of 1,2-dichlorobenzen was added and the reaction temperature was raised to 140~C for 3 h. Upon coolingto 0~C the title compound precipitated as a yellow solid which was collected by filtration and dried under vacuum. Yield 66.0 g. m.p. 215- 218~ C.

C. Preparation of compound of formula 39 25 4-N-(4-N.N-Dimethylaminomethylphenyl)amino-5-phenyl-pyrrolo~2.3-dlPyrimidine This compound was made by a route similar to Example 39B. Here, - the fluoroaniline was replaced with 4-N,N-dimethylaminomethylaniline. m.p. 208-209~ C.

D. Preparation of compound of formula 39 5-Phenyl-4-phenylaminopyrrolor2.3-dlPyrimidine This compound was made by a route similar to Example 39B. Here, the fluoroaniline was replaced by aniline. m.p. 208-209~ C.
E. Pre,u~, cflion of compound of formula 39 4-N-(4-Fluorophenyl)amino-5-(4-fluorophenyl)pyrrolo~2.3-dlPyrimidine This compound was made by a route similar to 70A, replacing phenacyl chloride by 4-fluorophenacyl chloride, followed by ~, eal" ,ent with 4-fluoroaniline as in example 70B; m.p. 245-248 ~C.

F Preparation of compound of formula 39 4-N-(4-Chlorophenyl)amino-5-phenylpyrrolo~2.3-dlpyrimidine This compound was made by a route similar to 70B by replacing 4-fluoroaniline by 4-chloroaniline.

G P~epar~lion of compound of formula 39 15 4-N-~4-(2-Hydroxyethyl)phenyl]amino-5-phenylpyrrolo[2.3-dlpyrimidine This compound was made by a route similar to 70B by replacing 4-fluoroaniline by 4-aminophenethyl alcohol; m.p. 206-208 ~ C

H Preparation of compounds of formula 42 Heterocycles as shown in scheme 15, were made accorcJing to the procedures 20 described in Browne et al. U.S. Patent Application Serial No. 081812/916.

I. Preparation of compound of formula 39 4-N-[(1.1.2-Trimethylpropvl)dimethylsilyloxymethyl]phenylarnino-5-phenylpyrrolo[2.3-dl~yrimidine The title compound was prepared by a route similar to 70B by replacing 4-25 fluoroaniline by 4-aminobenzyl alcohol followed by silylation with dimethyl thexyl chlorosilane; Rf 0.5 (silica, hexanes/ethyl acetate 50/50).

Example 84 Representative C~'-Svmmetrically Substituted Pyrrolo Pyrimidine Nucleosides Representative compounds of the invention, which can be made accordi"y to the methods described above, are identified in the following tables. With 5 r~rere, Ice to Formula 1, p, ~re" ~d compounds of the invention are pyrrolo pyrimidines (Y is carbon) where the A and B substituents are the same.
In one group of preferred compounds A and B are both HOCH2; in another, A and B are both hydrogen. G is preferably hydrogen, and E is preferably hydrogen or bromine, most ,orerer~bly hydrogen. Z, and Z2 are pr~rer~bly hydrogen 10 or methyl, most preferably hydrogen.
F

NJ~ ~D

~ N~N' 'E
B~o~

C 4'-SYMMETRICALLY SIJ~S l l l ~JTED PYRROLO PYRIMIDINE NlJCLEOSlDES

Y=C E=G=Z1 =Z2 =H

15 A=B=HOCH2 A = B = F D

H

27 phenylamino phenyl 2 28 4-fluorophenylamino phenyl 3 29 4-fluorophenylamino 4-fluorophenyl 4 30 4-fluorophenylamino 4-hydroxyphenyl 20 5 31 4-hydroxyphenylamino phenyl 6 32 4-hydroxymethylphenylamino phenyl 7 33 4-fluorophenylamino 3-pyridyl 8 34 3-pyridylamino phenyl 9 35 phenylmethylamino phenyl 25 10 36 4-(dimethylaminomethyl) phenylamino phenyl C~ SYMMETRICALLY StJ~ ~ JTED PYRROLO PYRIMIDINE NUCLEOClnFS
Y=C E=G=Z~ = Z2 =H
15 A=B=HOCH2 A=B= F D
11 37 4-(N,N-diethylethylenediamino phenyl ethyl)-phenylamino 12 38 4-fluorophenylamino 4-(2-dimethyl.. , ,oeU,yl)phenyl 13 39 4-cyanophenylamino phe.nyl 14 40 4-fluoruphe,,ylamino 4-cyanophenyl 41 4-(2-dimethylaminoethyl phenyl carbamoyl)phenylamino 16 42 4-(4-morpholinomethyl)phenyl amino phenyl 17 43 4-(2-(1-piperazino)ethyl)phenylamino phenyl 18 44 4-fluoruphe,,ylamino 4-(1-piperazinomethyl)phenyl 19 45 4-(sulrunal, o)phenylamino phenyl 46 4-(N-trifluoromethanesulfonyl phenyl amino)-phenylamino 21 47 4-fluoruphel,ylamino 4-(N-trifluoromethanesulfonylami no) phenyl 22 48 4guanidinophenylamino phenyl 23 49 4-fluorophenylamino 4-guanidinophenyl 24 50 4-(guan' ,o" ,ethyl)phenylamino phenyl 1 5 25 51 4-a" ,ophenylamino phenyl 26 52 4-fluorophenylamino 4-arnidinophenyl 267 299 4-cl.' ~ ruphenylamino phenyl 268 300 amino iodo 269 301 4-(2-(4- phenyl ",oruh-l ,o)ethyl)phe"~la" ,o 270 302 4-fluorupher,yl 4-(2-(1- -r~",o)ethyl)phenyl 271 303 4-(diethylaminomethyl)phenylamino phe 1yl 272 304 4-(1-r.-r;lZ",omethyl)phenylamino) phenyl 273 305 4-(1-piperidinoethyl)phenylamino phenyl 274 306 4-(1 -pyrrolidinoethyl)phenylamino phe 1yl 2~; 275 307 4-(diethylcarLo~dl";lo)phenylamino phenyl 276 308 4-(guar ,ocarbonyl)phenylamino phenyl 277 309 4-(2guanidinoethyl)phenylamino phenyl C-4'-SYMMETRICALLY SUt~S l l l UTED PYRROLO PYRIMIDINE NUC! FOclnF.c Y=C E=G=Z~ =Z2 =H
1 5 A=B=HOCH2 A= B = F D

278 310 4-(1-~. ~ d~i,.oacetyl)pher"~la" ,o phenyl 279 311 4-(gual ,oact:lyl)phenylamino phenyl 28Q 312 4-(2-(1-l~l,d~oiyl)ethyl)phenylamino phenyl 281 313 4-fluoropher"llamino 4-(5-l~l,d~olylmethyl)phenyl 282 314 4-(1-triazolylmethyl)phenylamino phenyl 283 315 4-(1 -i" Iylmethyl)phenylamino phenyl 284 316 4-mt:U,o~,-henylamino phenyl 285 317 4-ethùxy~,her"rlamino phenyl 286 318 4-(methylethyl)oxyphenylamino phenyl 1 0 287 319 3-methoxyphenylamino phenyl 288 320 4-fluoruphe, "~lamino 4-methoxyphenyl 289 321 4-fluo, upher,~lamino 3-ethoxyphenyl 290 322 4-l~elhuxyphenylamino 4-methoxyphenyl 291 323 4-ethoxyphenylamino 3-methoxyphenyl 1 5 292 324 3 4-methylenedioxyphenylamino phenyl 293 325 4-ethoxyphenylamino 4-methylphenyl 294 326 4-methylphenylamino phenyl 295 327 3-ethylaminophenylamino phenyl 296 328 4-(methylethyl)phenylamino phenyl 297 329 4-fluorophenylamino 4-methyl 298 330 4-fluorophenyl 3-guar ~ophenyl 447 467 3,4-dimethoxyphenylamino phenyl 448 468 3,4,5-trimethoxyphenylamino phenyl 449 469 4-fluorophenylamino 3,4-dimethoxyphenyl 450 470 4-fluorophenylamino 3 4-ethylenedioxyphenyl 451 471 4-(2-methoxyethyloxy)phenylamino phenyl 452 472 3-ethoxy-4-fluorophenylamino phenyl W O 96/40705 PCTrUS96/10404 C 4'-SYMMETRICALLY SU~ .)TED PYRROLO PYRIMIDINE NUCLEOSIDES

Y=C E=G=Z, =Z2 =H
A=B=HOCH2 A=B=H F D
453 473 3-al l lophe"~lamino phenyl 454 474 3-amino4-fluo,uphe,lylamino phenyl 455 475 4-fluorupher,ylamino 3-dl, llophe,."l 456 476 3-ethyb., ~ ,ophe"yldl, ,o phenyl 457 477 3-diethyh" ,;. ~uphe, ,ylamino phenyl 458 478 4-fluc.,upher"/lamino cyclohexyl 459 479 5-be"~i" Ny lamino phenyl 460 480 4~2- phenyl diethyld", ,ot:Ll "~lo)gr)phenylamino 461 481 4-(2-carboxyethyl)phenylamino phenyl 462 482 4~2-carboxyethenyl)~,her"rld" ,o phenyl 1 0 463 483 4-carboxyphenylamino phenyl 464 484 4-fluo, upher,yl 4-(2-~, bùxyethenyl)phenyl 465 485 4-(carboxymethyl)pher,yld", .o phenyl 466 486 4-(2-N- phenyl aminoguan ,o~:Ll ,yl)phenylamino When E is bromine and A and B are both HOCH2 a p, t r~ u~d compound is one where 1 5 (53) F is 1 fluuruphenylamino and D is phenyl. Using the same d~ri, 'i~ ns for D E F G and Z, . Z2.
another prere.,~d compound is one where (54) A and B are both hydrogen.

~Xdl l r ~ 76 Re~r~senlaLi~re C4' Uns~""",~l,~ -~Iy Suhstihlt~d Pyrrolo Pyrimidine NU~ OC;~ OS
Other prefered pyrolo pyrimidine compounds of the invention are those where A and 20 B are not the same as shown in Tables 2 and 3.

C 4'-UNSYMMETRICALLY SlJ~5~i 111 UTED PYRROLO PYRIMIDINE NUCLEOSIDES
Y=C E=G=Z, =Z2 =H
A=HOCH2 B=HOCH2 F D
B=CH3 A=CH3 81 phenylamino phe!nyl ~UBST!TUTE SHEET (RU~F 26~

C 4'-UNSYMMETRICALLY SUBSTITUTED PYRROLO PYRIMIDINE NUCLEOSIDES
Y=C E=G=Z~ =Z2 =H
A=HOCHz B=HOCH2 F D
B=CH3 A=CH3 56 82 4-fluorophenylamino phenyl 57 83 4-fluorophenylamino 4-fluorophenyl 58 84 4-fluorophenylamino 4-hydroxyphenyl 59 85 4-hydroxyphenylamino phenyl 86 4-hydroxymethyiphenylamino 4-fluorophenyl 61 87 4-fluorophenylamino 3-pyridyl 62 88 3-pyridylamino phenyl 63 89 phenylmethylamino phenyl 64 90 4-(dimethylaminomethyl) phenyl phenylamino 1 0 65 91 4-(N,N-diethylethylenediamino phenyl ethyl)phenylamino 66 92 4-fluorophenylamino 4-(2-dimethylaminoethyl)phenyl 67 93 4~yanophenylamino phenyl 68 94 4-fluorophenylamino 4~yanophenyl 69 95 4-(2-dimethylaminoethyl phenyl carL al l luyl)-phenylamino 96 4-(4-morpholinomethyl)phenyl amino phenyl 71 97 4-(2-(1-piperazino)ethyl)phenylamino phenyl 72 98 4-fluorophenylamino 4-(1-piperazinomethyl)phenyl 73 99 4-(sulr~ndlll :'o)phenylamino phenyl 74 100 4-(N-trifluoromethanesulfonyl phenyl amino)-phenylamino 101 4-fluorophenylamino 4-(N-trifluoromethane-sulfony I amino)phenyl 76 102 4-guanidinophenylamino phenyl 77 103 4-fluorophenylamino 4-guanidinophenyl 78 104 4-(guanidinomethyl)phenylamino phenyl 79 105 4-amidinophenylamino phenyl 106. 4-fluorophenylamino 4-amidinophenyl When E is bromine, A is HOCH2 and B is CH3, a preferred compound is one where (107) F is 4-fluorophenylamino and D is phenyl. Using the same definitions SUBSTlTUT~ SHEET (RUI~ 26) W O 96/40705 PCT~US96/10404 for D, E, F, G, and Z,. Z2. another preferred compound is one where (108) A is CH3 and B is HOCH2.

ExamDle 77 Additional C~' Unsymmetrically Substituted Pyrrolo Pyrimidine Nuclesides Still other prefered pyrolo pyrimidine compGunds of the invention are those where one of A and B is CH3 and the other is C~1~1~12, or one of A and B is methoxymethyl or CH20H, or one of A and B is C~ OH and C~ Nl~ as shown in Table 3.

1 0 MORE C 4'-UNSYMMETRICALLY SIJ~ I I I UTED PYRROLO PYRIMIDINE
NUCLEOS!nF-S
Y = C E = G = Z1 = Z2 = H
A=CHs A=H2NCH2 F D
B=H2NCH2 B=CH~
1~ 109 128 phenylamino phenyl 110 129 4-chloruphenylamino phenyl 111 130 4-fluoruphenylamino 4-fluc,ruphen~rl 112 131 4-fluorophenylamino 4-hydroxyphenyl 113 132 4-hydroxypher,~la., ~ ,o phenyl 114 133 4-hydroxymethylphen~la",, phenyl 115 134 phe"~lau ,o 3-pyridyl 116 135 3-pyridylamino phenyl 117 136 phenylmethylamino phenyl 118 137 4-cyanophenylamino phenyl 2~i 119 138 4-fluo, upllen~rlamino 4-cyanophenyl 120 139 4- carbamoylphenylamino phenyl 121 140 4-(sulfonamido)phenylamino phen~tl 122 141 4-(N-trifluoromethanesulfony phenyl I amino)phenylamino 123 142 phenylamino 4-(N-l rifluorui "t:U ,~nesulfonylamin o) phenyl 124 143 4guanidinophenylamino phenyl 125 144 4-fluorophenylamino 4guar ,opl el,~l 126 145 4-a". ,ophenylamino phenyl 1 0 MORE C 4'-UNSYMMETRJCALLY Sl 1~ ITED PYRROLO PYRIMIDINE
NUCLEOSIr~FS
Y=C E=G=Z~ =Z2 =H
A=CH~ A=H2NCH2 F D
~ B=H2NCH2 B=CH~
127 146 4-fluorophenylamino 4-dlN ,opher,ll 331 332 4-fluorophenylamino phenyl A=CH~OCH A=HOCH2 F D
2 B=HOCH2 B=CH ,OCH2 5 333 353 phenylamino phenyl 334 354 4-chlorophenylamino phenyl 335 355 4-fluorophenylamino ~fluo,uphe,,~l 336 356 4-fluorophenylamino 4-hydroxyphenyl 337 357 4-hydroxyphenylamino phenyl 1 0 338 358 4-hydroxymethylphenylamin phenyl o 339 359 phenylamino 3-pyridyl 340 360 3-pyridylamino phenyl 341 361 phenylmethylamino phenyl 342 362 4-cyanophenylamino phenyl 1 5 343 363 4-fluorophenylamino 4~rdnophe"~1 344 364 4- carbamoylphenylamino phenyl 345 365 4-(sulrùna", ~ ~)phenylamino phenyl 346 366 4-(N-trifluorom~:~ ,anesulfony phenyl I amino)phenylamino 347 367 phenylamino 4-(N-tr~fluoromethanesulrur,~la",, o) phenyl 348 368 4~uanidinophenylamino phenyl 349 369 4-fluorophenylamino 4~uan ,ophe"~l 350 370 4-amidinophenylamino phenyl 351 371 4-fluorophenylamino 4-amidinophenyl 352 372 4-fluorophenylamino phenyl A=H2NCH2 A=HOCH2 F D
B=HOCH2 B=H2NCH2 373 393 phenylamino phenyl 374 394 4-chlorophenylamino phenyl 375 395 4-fluorophenylamino 4-fluoruphe~
376 396 4-fluorophenylamino 4-hydroxyphenyl CA 02220642 l997-ll-28 WO 96140705 PCTrUS96/10404 25 A=H2NCH2 A=HOCH2 F D
B=HOCH2 B=H2NCH2 377 397 ~hydrox~,her.yld,. .o phenyl 378 398 4-hydroxymethylphenylamin phenyl 379 399 pher"rla~ 3-pyridlyl 380 400 3-p~ Jylar, ~o phenyl 381 401 phenylmethylamino phenyl 382 402 4~y~nopl e.. ylamino phenyl 383 403 ~fluorupher.~rld.. .o 4-c~dhophell~l 384 404 4- carLd....... uylphenylamino phenyl 385 405 4 (sl~lr~.r.d.. ..-)phenylamino phenyl 10 386 406 4-(N-tnfluoru.. e:ll.anesulfony phenyl I amino)phenylamino 387 407 pher.yld....... .o ~(N-trifluorom~:ll,dl~e~.llfonylamin o) phenyl 388 408 4 guanidinophenylamino phenyl 389 409 4-fluorupl el"llamino 4~ual ,oph~yl 390 410 ~ar, ,opher.,~la.. .o phenyl 15 391 411 ~fluoruphe"~lamino 4-all .ophenyl 392 412 4-fluorophenylamino phenyl When E is bromine, A is NH2CH2 and B is CH ~, a prerer, ed compound is one where (147) F is 4-fluorophenyiamino and D is phenyl. Using the same definitions for D, E, F, G, and Z1, Z2~ another ,." ~r~" ed compound is one where ~148) A is CH3 and B is NH2CH2.

Example 78 Representative C~' Spirocyclic Pyrrolo Pyrimidine Nucleosides A and B can together form a cyclopropyl ring. Prefered pyrolo compounds of this kind, where E, G, Z, and Z2 are all hydrogen, are shown in Table 4.

TABLE 4 C-4' SPIROCYCLIC PYRROLO PYRIMIDINE NUCLEOSIDES
A & B FORM A RING Y = C E = G = Zl = Z2 = H
F D
149 phenylamino phenyl 150 4-fluorupher.yL.. .o phenyl W O 96/40705 PCT~US96/10404 TABLE 4 C-4' SPIROCYCLIC PYRROLO PYRIMIDINE NUCLEO~:!r)F-S
A&BFORMARING Y=C E=G=Z~ =Z2 =H
F D
151 4-fluorophenylamino 4-fluol upher~l 152 4-fluGruphenylamino 4-hydroxyphenyl 153 4-hyd~ her,ylamino phenyl 154 4-hydroxymethylphenylamino 4-fluoruphe,, 155 4-fluorophenylamino 3-pyridyl 156 3-pyridylamino phenyl 157 phenylmt~ ld" ~o phenyl 158 4-(d "eU,Jla., .u,,,ethyl)phenylamino phenyl 159 4-(N,N-diethylethylenediaminoethyl)phenyl phenyl amino 10 160 4-fluoropheh~lar, ,o 4-(2-dimethylan ,ot:ll"rl)phenyl 161 4-cyanophenylamino phenyl 162 4-fluorophenylamino 4-cyanophenyl 163 4-(2-dimethylaminoethylcdrl ar"u~l)- phenyl phenylamino 164 4-(4-morpholinomethyl)phenylamino phenyl 15 165 4-(2-(1-~. rdZ,-,o)ethyl)phenylamino phenyl 166 4-fluorophenylamino 4-(1 - . - rd~"omethyl)phenyl 167 4-(sulrundl lo)phenylamino phenyl 168 4-(N-trifluoromethanesulfonylamino)phenyl phenyl amino 169 4-fluorophenylamino 4-(N-trifluoromethanesulfonylamino) phenyl 20 170 4-guanidinophenylamino phenyl 171 4-fluorophe"~la.1 ,o 4-guanidinophenyl 172 4-(guanidinomethyl)phenylamino phenyl 173 4-all ,opl,enylamino phenyl 174 4-fluorophenylamino 4-amidinophenyl 25 175 amino iodo TABLE 4a C-4' SPlROA~t ~ INO PYRROLOPYRIMIDINE NUCLEOSIDES
Y=C E=G=Z~ =Z2 =H
A ~ B Forrn a ring F D
30 containing a nitrogen 413 phenylamino phenyl 414 4-fluorophenylamino phenyl TABLE 4a C 4' SPlROA~t ~ lO PYi~ROLOPYRlMlDlNE NUCLEOSIDES
Y=C E=G=Z~ =Z2 =H
A & B Forrn a ring F D
30 co~ -,g a l.it,~,gen 415 4-fluorophenylamino 4-fluorophenyl 416 4-fluoropher"rla" ' ,o 4-hydroxyphenyl 417 4-hydroxyphenylamino phenyl 418 4-hydroxymethylphenylamino phenyl 419 phenylamino 3-pyriclyl 420 3-pyridylamino phenyl 421 phenylmethylamino phenyl 4~ 4~yanophenylamino phenyl 423 4-fluorophenylamino 4~yanophenyl 424 4- cd,l,d.,.oylphenylamino pheny'l 425 4-(sulruna",''~)phen~la,., ,o phenyl 426 4-(N-tn'fluoromethanesulfonyl phenyl amino)phenylamino 427 phenylamino 4-(N-bifluoromethanesulfonylamin o) phenyl 428 4guanidinophenylamino phenyl 1 5 429 4-fluorupher.~lamino 4gua, ' ' ,ophenyl 430 4-amidinophenylamino phenyl 431 4-fluorophenyiamino 4-al, ~ ,ophenyl When E is bromine, and A and B form a cyclopropyl ring, a preferred compound is one where (176) F is 4-fluorophenylamino and D is phenyl.

20 Example 79 Representative C-4' Symmetrically Substituted Pyrazolo Pyrimidine Nucleosides An acldiLio"al group of prefered compound are the pyrazolo pyrimidines, where Y is nitrogen and E is nothing. Representative gem pyrazolo compounds, where A and B are the same (in this case both hydrogen) are shown in Table 5.

W O 96/40705 PCT~US96/10404 C ~' SYMMETRICALLY SUBSTITUTED PYRAZOLO PYRIMIDINE NUC! FOSlr~ES
Y=N G=Z, =Z2 =H
A=B=H F D
177 phenylamino phenyl 178 4-fluoruphen~lamino phenyl 179 4-fluorupher.~lamino 4-fluorophenyl 180 4-fluorophenylamino 4-hydroxyphenyl 181 4-hydroxyphenylamino phenyl 1 0 182 4-hydroxymethylphenylamino 4-fluorophenyl 183 4-fluorophenylamino 3-pyridyl 184 3-pyridylamino phenyl 185 phenylmethylamino phenyl 186 4-(dimethylaminomethyl)phenylamino phenyl 1 5 187 4-(N,N-diethylethylene.lidn ,oethyl) phenyl phenylamino 188 4-fluorophenylamino 4-(2-dimethylaminoethyl)phenyl 189 4-cydnophe--~ - ~o phenyl 190 4-fluorophenylamino 4-cyanophenyl 191 4-(2-dimethylaminoethyl phenyl carbamoyl)-phenylamino 192 4-(4-rllul~h~ ~o,--ethyl)phenylamino phenyl 193 4-(2-(1 ~r ', d~il ,o)ethyl)phenylamino phenyl 194 4-fluorophenylamino 4-(1-piperazinomethyl)phenyl 195 4-(sulrunat. o)phenylamino phenyl 196 4-(N-trifluoru,--eU,anesulfonylamino) phenyl phenylamino 197 4-fluorophenylamino 4-(N-trifluoromethanesulfonylamino) phenyl 198 4-guan ,ophen~lamino phenyl 199 4-fluorophen~lan ,o 4-gual ,ophenyl 200 4-(9Udl .or.. ethyl)phenylamino phenyl ~ 201 4-amidinophenylamino phenyl 30 202 4-fluorophenylamino 4-al . .opher ~rl ~ 432 pher.~rla,- ~o 4-chlorc,phenyl 433 4-methoxyphenylamino phenyl 434 4-ethoxyphenylamino phenyl 435 4-(methylethyl)oxyphenylamino phenyl C 4' SYMMETRICALLYSUBSTITUTED PYRAZOLO PYI;~ INE ~I~!C! FOSI~F-C
Y=N G=Z, =Z2=H
A=B=H F D
436 3-methoxyphenylamino phenyl 437 4-fluoropher,~ld", ,o 4-",t:llw~l,har,~
438 4-fluorophenylamino 3-ethoxyphenyl 439 4-~ II-uxyphenylamino 4-methoxyphenyl 440 4-ethoxyphel,~rlc.. , lo 3-methoxyphenyl 441 3,4-methylen_ yphenylamino phenyl 442 4-ethoxyphenylamino 4-methylphenyl 443 4-methylphenylamino phenyl 444 3-ethylaminophenylamino phenyl 1 0 445 4-(methylethyl)phenylamino phenyl 446 4-fluorophenylamino 4-methyl Example 80 C~' Unsymmetrically Substituted Pyrazolo Pyrimidine Nucleosides Still other prefered pyr~7010 pyrimidine compounds of the invention are 15 those where one of A and B is CH3 and the other is H2NCH2, as shown in Table 6.

C 4' UNSYMMETRICALLY SUBSTITUTED PYRAZOLO PYRIMIDINE
NUCLEOSIDES
Y= N G=Z~ = Z2 = H
A=CHa A=H2NCH F D
B=H2NCH 2 B=CHa 203 2~ phenylamino phenyl 204 223 4-fluorophenylamino phenyl 205 224 4-fluorophenylamino 4-fluorupher,yl 206 Z5 4-fluorophenylamino 4-hydroxyphenyl 207 Z6 4-hydroxyphenylamino phenyl 208 227 4-hydroxymethylphenylamino phenyl CA 02220642 l997-ll-28 C~' UNSYMMETRICALLY SlJ~S ~ ITED PYRAZOLO PYRI~înlNC
~ NUCLEOSIDES
Y = N G = Z~ = Z2 = H
A=CH~ A=H2NCH F D
B=H~NCH 2 B=CHS
209 228 4-fluorophenylamino 3-pyridyl 210 229 3-pyridylamino 4-fluorupher.
211 230 phenylmethylamino phenyl 212 231 4-cyanophenylamino phenyl 213 232 4-carbamoylphenyiamino phenyl 214 233 4-fluoruphe, ~ylamino ~cyanopher,~l 215 234 4-(sulrona,.,''~)phenylamino phenyl 216 235 4-(N-trifluor~,n,ell,anesulfonyl phenyl amino)-phenylamino 217 236 4-fluorphenylamino 4-(N-trifluoromethanesulfonylamin o) phenyl 218 237 4guar,' ' ~ophenylamino phenyl 219 238 4-fluorophenylamino 4~uar' ' ~ophenyl ~o 239 4-a.. ' " .ophenylamino phenyl 221 240 4-fluorophenylamino 4-amidinophenylamino Example 81 15 Representa~ive C4' Spirocyclic Pyrazolo Pyrimidine Nucleosides A and B can together form a cyclopropyl ring. Preferred pyrazolo pyrimidine n~lcleosides of this kind, where G, Z, and Z2 are all hydrogen, are shown in Table 7.
.

TABLE 7 C~' SPIROCYCLIC PYRAZOLO PYRIMIDINE NUO! FOS!nES
A&BFORMARING Y=N G=Z~ =Z2 =H
F D
241 phenylamino phenyl 242 4-fluorophenylamino phenyl 243 phenylamino, 4-chlororophenyl 244 4-fluoruphe.. ~lamino 4-hydroxyphenyl 2~ 245 4-hydroxyphenylamino phenyl 246 4-melh~ ,,hen~lamino phenyl W O 96/40705 PCT~US96/10404 TABLE 7 C 4' SPIROCYCLIC PYRAZOLO PYRI~lnlN~ NUCLE~ClnF~:
A&BFoRMARlNG Y=N G=Z1 =Z2 =H
F D
247 4-hydroxymethyiphenylamino phenyl 248 4-fluorophenylamino 3-pyridyl 249 3-pyridylamino phenyl 250 phenylmethylamino phenyl 251 4-~ .. ell.~rl.. o.. t:U.~l)phenylamino phenyl 252 4-(N,N-diethyl~U-Jlenediaminoethyl) phenyl pher,~ld...' .o 253 4-fluo.~,pher.~rlamino 4-(2-dimethyJai, ~ .ot:U.~l)phenyl 254 4-cy~nophe.-~lamino phenyl 255 4-fluorophenylamino 4-cyanophenyl 256 4-(2-dimethylaminoethylcdrLd---oyl)- phenyl pher,JI~
257 4-(~mo,~h~~ lor"ethyl)phenylamino phenyl 258 4-(2-(1-r -rd~i,.o)ethyl)phenylamino phenyl 259 4-fluoruphell~rldlll .o 4-(1- -rd~i.. omethyl)phenyl 260 4-(sulr~,nar. ~ Ic)phenylamino phenyl 1 5 261 4-(N-trifluoromethanesulfonylamino) phenyl phenylamino 262 4-fluorophenylamino ~1 (N-trifluoromethanesulfonylamino) phenyl 263 4-gu-du~ ,ophe"ylamino phenyl 264 4-fluorophenylamino 4 guanidinophenyl 265 4-(guan ,o",t:U"~l)phenylamino phenyl 266 4-drl l ,ophenylamino phenyl UTILITY
The adeno-~ii"e kinase inhibitors of the present invention may be used in the treatment of a variety of clinical situations where increasing local levels of ade"osi"e are beneficial. The compounds of the invention act as potent inhibitors 25 of adenosine kinase in vitro and the present compounds in particular are orally available.
Adenosine has been proposed to serve as a natural anticonvulsant.
Compounds of the present invention which enhance adenosine levels are useful in seizure disorders as shown in animal models of seizures de~ailed below. Adenosine 30 kinase inhibitors may be used in the treatment of patients ~rith seizures or epilepsy .

W O 96/40705 PCT~US96/10404 or patients who might have chronic low or insufficient adenosine levels or mightbenefit from increased adenosine such as those suffering from autism, cerebral ~ palsy, iI ,so" " ,ia or other neuropsychiatric s~," IptOI, .s.
Adenosine kinase inhibitors of the invention find further utility in the 5 treatment of acute pain, including but not limited to peri-operative, post-surgical, and end-stage cancer pain. Compounds of the invention are also useful in controllingcl "."~ic pain, including but not limited to pain c~used by arthritis, cancer, trigeminal neuralgia, multiple scierosis, neuropathies such as those arising from ~ hetes and AIDS and in addition, lower back pain and phantom limb pain. Treatment of acute 10 and cl1r~".ic pain can be treated by ad~ ,isl,~lio,- of the compounds of the invention in a systemic or oral fashion, as illustrated by animal models detailed below.
Adenosine has been reported to be an endogenous modulator of inflammation by virtue of its effects on stimulated neutrophil function and on ",a~ u~hage, Iymphocyte and platelet function. The compounds of this invention may 15 therefore be used in treating conditions in which inflar"",aLo~y processes are prevalent such as a, lhIilis, reperfusion injury, and other inflan,",alory disorders.
The compounds of the invention are also useful in the treatment of chronic neurodegenerative r~ise~se, such as Alzheimer's dise~se, Parkinson's disease, ALS, Huntington's disease, and AIDS dementia.
Stroke and central nervous system ("CNS") trauma are conditions where tissue injury results from reduced blood supply to the CNS and are thus amenable to an intervention that provides increased levels of adenosine to the compromised tissue. It is reported that a significant com~.o"ent of the neurodegeneration resulting from stroke or CNS trauma is caused by increased excitatory amino acid release and sensitivity, which results in neurons being stimulated to death. In addition to vasodilatory properties, adenosine has been reported to inhibit release of excitatory amino acids (Burke and Nadler J.
Neurochem., 1988, 51:1541) and responsiveness of neurons to excil~liol1. The compounds of this invention, which increase adenosine levels, may also be used in the Lr~LInenl of conditions where release of or sensitivity to exciLa~o, y amino acids is implicated.

CA 02220642 l997-ll-28 To assist in ~ Id"ding the present illl,~..liùl~5 and especially their ~Iu~-~llids and utilities, the results of a series of ~A~ are also included. These eA~ lllull~lldl~l that a number of c~ M~ of the present invention were potent inhibitors of ~ purified cardiac ~ ?
kinase. Certain a Ir~ o kinase inhibitors were found to inhibit seizures and exhibit anti i~r~ .y activity in well-~ -d animal models. The results of these eA~ ,i",~ ; are shown in Table 8.
TABLE 8 UTlLlTY OF Re~ e~eNIATlVE COMPOUNDS
# Name (ICso) AK Call _ Paw 1%inh) A
Inhibition Aotivity (MESI
~nM) P~ i-P i.p. (mglkg) ~ Alph-~ ~ 5-phenyl-7-14-C
h, ' u,~ hyl 1,B;D; ., 1.2 18 3 > 1.00 " .),,.1- . 3~1,"
(m p. 232 C) 1 0 150 4N (4-F' " ' , !; 5-phenyl-7-(4-C- . I , ~ r ~l ~ D~ 0 3 80 78 0.23 [2,3 ']~ ,. " (m.p. 142 C) 175 4-Amino-5-iodo-7-(4C-~;yl;lll' ': ;I''7 3 600 l, ,. " (m.p. 232 C) 81 4-11/-r; ,: 5-phenyl-7-(4-C-methyl-B-D-.il,~u. ,:; ~.,.u ' '? ~- 2000 ~Jrl.i (m.p. 211-213 C) 64 4-A/-14-(r ' ,~
~.. _lh~'), ,!, 5-phenyl-7-(4G 70 >5 0 methyl-B-D l~,.u0.~ ".~.' '? 3-d]~ -,;,lh,e (m.p. 205-206 C~
P; ,' 5-phenyl-7 (,B-D-27 ~,y~hll,f~ ".. ' '? ~- 4 12 44 <5.0 '], "i ' (m.p. 210-211 C~
~1 /V (q F , ' ,I: ~ - 5 phenyl-372 7 (4 C III~IIIUA~. hyl ,8 D 24 > 5.0 I~Au~u-a"u~ r"-ulù[Z3-d]~"illlillill~ (m.p. 205-206 Cl rl~.~.,.."h_..,l) ~ - 5 phenyl-352 7;(4-C ' ~ ~yl b-D- 400 a~ùi~.,r~.lulu~2,3 .J]".illliJlllr, (m.p. 198-199 C~

SU8STITUTE SHEET (RULE 261,!

W O 96/40705 PCTnUS96/10404 TABLE8 UTlLln OF~Lr~c~tNlATlVECOMPOUNDS
# Name (ICso~ AK Cdll Paw (~inh) A
- InhibitiDn ActiYity (MES) (nM) P-~- i.p. i.p. (mglkg) I A (~ Flh~ 5-phenyl-7-(4-C- ~ :b~: b -D-392 L , )".,1 3:l" ~ 1100 -b~3~Imp. 200-220 C
dec) 3-(4-C . ~1)1-lb-D-432 ~lylllll ,: ~/1/ 9O 15 24 >5.0 (m.p. 194-195 C) 4-/Y-(4-Fluo,. ,1 5-phenyl-28 7-(b-D-~y _ , ), ,.lu10l2,3- 28 25 52 <5.0 d] ,.i(m.p. 194195 C) 4-A~-(4r , ~ ,~; e 1~!
r .~.l ,:) 7-lb-D- 45 20 46 .,. ytlll ~
.11, ~,i ,idill~ Im.p. 200-202 C~
4JV(~Lr~, !; ~ 5phenyl-299 7-lb-D elytlllurl"~ , ~".II .2 3- 40 28 20 ]" ~ Im.p. 212-213 CJ
4-Amino-5 iodo-7-1b-D-300 ..yllllu~ ".1ulu[2,3- 6 91 90 r9]~ ,. Im.p. 258 C Idec.

, ~ 5 301 phenyl-7- 1 39 21 (b-D L.yllll~ ".,~ ' '7 3-~l]~ ~'- Im.p. 128-130 C~
4-Ar14-12-11-r~ , ~ 5~ ~, " ~ 5-43 phenyl-7-lb-D- 3.5 34 65 ~, y llll, ~ " 1 1 . 3 ~],, ~ 1I h~. n id~ Im-p-150-180 Idec.l~

SUBSTITUT~ SHEET (RULE 26) WO 96/40705 PCT~USg6/10404 AK INHIBITION
Adenosine kinase activity was measured esse"Lially as described by Yamada et a/. (Yamada, Y., Goto, H., Ogasawara, N. (1988) Bfochlm. Biophys. Acta660, 36~3.) with a few minor modifications. Assay mix~ures contained 50 mM TRIS-maleate buffer, pH 7.0, 0.1% BSA, 1 mM ATP 1 mM MgCI2, 0.5 ~M [U-14C]
adenosine (4û0~00 mCiimrnoij and varying dupiicate concentraiions of inhibitor.
The rt:ac~io"s were i, liLi~led by addilion of approximately 0.1 ~U partially purified pig heart adenosi. ,e kinase or recombinant human adenosine kinase (Spychala, J. et al., Proc.Natl.Acad.Sc~. USA 93, 1232-1237, (1996)), where one unit is defined as that 10 amount of enzyme required to phosphorylate 1 ,~lmol adenosine per minute. Thereactions were incl ~h~ted for 20 minutes at 37~ C. The assay was quenched upon spo~Ling 30 l~l aliquots onto 2 cm2 pieces of Whatman DE8 1 anion exchange paper.
The papersquares were washed for 3 minutes in 6 L distilled/deionized water to remove the u"l ea~;~ed adenosine. The washed squares were rinsed in 95% ethanol 15 and dried in an oven at 100~C for 10 minutes. The amount of '4C-AMP was quantified by sdntillation counting. The concentration of inhibitor required to inhibit 5û% of the adenosine kinase activity (IC50) was determined graphically. The results for representative adenosine kinase inhibitors of the invention are shown in Table 8.

ANTICONVULSANT ACTIVITY
The anticonvulsant activity of the tested compounds was evaluated in male SA rats (100-1509, Simonsen) using the maximal electroshock (MES) model described in Swinyard et al., Anfiepilepfic Drugs, 3d Ed. at 85-102 (Levy, et al., eds.), NY: Raven Press (1989). The rats were maintained on a 12/12 lighVdark cycle in temperature controlled facilities with free ~ccess to food and water. For p.o.
2~ administration, the animals are fasted overnight, prior to the experiment. One hour prior to seizure testing, the animals were injected interperitoneaily (ip) or orally (per os, po) with one of various doses of test compound dissolved in DMSO or PEG 400.
Maximal electroshock seizures (MES) were induced by administering a 150 mA, 60 Hz current for 0.2 seconds via corneal electrodes using a Wahlquist W O 96/40705 PCT~US96/10404 Model H stimulator. The e, IdpoinL measurement was suppression of hind limb tonic extension (HTE), which was judged to occur when any hind leg extension did not - ~yree~ a 90 degree angle with the plane of the body. HTE suppression of this kind - indicates that the test compound has the ability to inhibit seizures, in theory by 5 inhibiting seizure pr..pa~ ;o" and spread, if not by raising the seizure threshold (i.e.
preventing seizure potential). This endpoint was expressed as the per;e"lage of animals in which the response was inhibited. Typically, co",,l~ounds were screened initially at one hour following a dose of 5 mg/kg ip. In some cases, the effective dose at which 50% of the rats were protected (EDso) was ~ c~ ted from a dose response10 curve. The results for exemplary col"pounds of the invention are set forth in Table 8, ex~r~ssed as ED50 values. For compounds where the ED50 was not c~l~ ll~t~ , the result is listed as >5 if HTE was inhibited in fewer than 50% of the a, li" lals in the intial screen, or <5 if HTE was inhibited in more than 50% of the animals in the intialscreen.

ANTI-INFLAMMATORY ACTIVITY
Carrageenan (Type A) was suspended in sterile PBS at 1% (w/v), ~ ~orl~Yed for 30 minutes, and stored at room temperature. Rats were pretreated with vehicle or AK inhibitor (10 mg/kg) by oral gavage or i.p. administration and the volume of the left hind paw was measured using a water displacement 20 plethysmometer (Stoelting Co., Wood Dale, IL). One hour after oral treatment or 30 minutes after i.p. treatment, the rats were briefly anaesthetized, and 0.1 ml of the carrageenan solution was i, I;~cted s~ IhCUt~rleOUSly into the planar surface of the left hind paw. The ensuing paw swelling was measured by plethysmometry after 3 hours. The paw volume in milliliters was subtracted from the pre-injection paw 25 volume. Data are presented in Table 8 as the percent inhibition of paw swelling in AK inhibitor treated animals, compared to vehicle treated control animals.
Rosengren et al., J. Immunology 154: 5444-51 (1995).

LIVER TOXICITY
Fernale SA rats 5150-200 g) were anesthetized with halothane and 30 cannulated via the internal jugular vein. The animals were allowed to recover for 3 WO 96/40705 PCTnUS96/10404 days. At this time, 37.5 ,umol/kg of an AK inhibitor was dissolved in 75%
PEG400/25% saline and infused through the jugular catheter over 40 minutes.
Twelve hours later, an additional 37.5 ,umol kg was infused over 40 minutes (total dose = 75 ,umol/kg). Twelve hours after the second dose, the animals were 5 anesthetized with halothane and exsanguinated through the descending aorta.
Serum was prepared and liver enzymes (serum glutamic-oxaloacetic transaminase (SGOT), serum glutamic-pyruvic transaminase (SGPT)) and total bilirubin in ~he serum samples were determined by a co" ~" ,ercial laboratory. Results are shown in table 9.

1 0 TABLE 9: ~VERAGE SERUU CONTENT OF SGOT, SGPT AND TOTAL BILIRUBIN
SGOT SGPTTotal Bilirubin #;~ animals (U/mL) (U/mL) (UlmL) 2 65 53 0.1 150 2 560 155 1.58 27 5 114 41 0.14 28 4 61 23 0.07 29 4 130 30 0.16 43 5 99 58 0.1 RAT SKIN LESION MODEL
Rat skin lesions were induced as in Rosengren et al., J. Immunology 20 154: 5444-51 (1995). The dorsal skin of male SA rats was shaved and carrageenan (Type A) or phos~,hale-buffered saline was injected intradermally. Three hours later, the injection sites were b.opsied and weighed. Neutrophil conlenL of the skin biopsies was measured as the quantity of myeloperoxidase (MPO) present in a tissue homogenate. The .oyrise~i~ weighed skin pieces were placecl in 4 ml 0.5% mixed alkyl 25 Ll i" ,e~l ,ylam" ,o"ium bromide and homogenized at the highest speed for 15 seconds in a Polytron homogenizer (Brinkmann Instruments, Westbury, NY). Lipids were extracted by adding 1 ml of dichloromethane to the homogenate, vortexing W O ~10/~ PCTAUS96/10404 ~,;yuluu~ly, and centrifuging at 1000g, 5~C for 15 min. Fifty ~1 of each Su~ lldldlll was added in duplicate to a 96 well assay plate, along with dilutions of human ul~r~lo~e~oAidase standard. Puld~;~;uul ~Jhu;~Jlldl~ buffer (pH 6.1) containing 0.36 mglml of o-dianisidine dihyd~- '' iJe and 0.001% hydrogen 5 peroxide was added t200 ~11well), and the al,su-l.dnce at 450 nm was read after 5 min ;-~ h ,liun at room I~ dlul~. The myelope-uAidase content of each skin piece was calculated from a standard curve cu~ dd using least-square l~yl~;ùn, and ~A~ ssed as units of MPOlg of tissue.
AK inhibitors were administered orally using poly~.lllyl~ glycol-400 as vehicle or i"L,d~.~,ilo"eally using dinlelll~b~ uAide as vehicle, at indicated time before skin lesion injection. For each eA~ lll, the 10 average of all values from saline-induced lesions was calculated. This baseline value was then S~blld~ d from values obtained from Cd.ldy~ud', induced lesions. Percent inhibition for AK inhibitors were calculated from these baselinc c...,-,~t~d values. The results are shown in Table 10.

TABLE 10: Inhibition of ~ in rat skin lesions induced by G.--l _ F~ Time ## animalsRoute Dose (mglkg)% inhibition SEM
(min) 1 5 27 5 i.p. 3û 3 2 14 27 5 i.p. 30 10 59 7 27 5 i.p. 30 30 89 2 27 8 i.p. 60 10 60 4 27 4 i.p. 120 10 14 6 27 4 i.p. 180 10 2 26 27 4 i.p. 240 10 0 27 4 i.p. 360 10 0 28 4 i.p. 30 3 18 10 2812 i.p. 30 10 62 5 28 4 i.p. 30 30 79 11 28 8 i.p. 60 10 49 7 28 8 i.p. 120 10 60 4 SUBSTITUTE SltEET (RULE 26) CA 02220642 l997-ll-28 28 8 i.p. 120 10 60 4 28 8 i.p. 240 10 28 8 28 8 p.o. 60 10 28 11 28 25 p.o. 60 20 40 7 28 10 p.o. 120 20 84 4 28 8 p.o. 240 20 77 4 28 8 p.o. 420 20 20 18 29 4 p.o. 60 10 0 29 4 p.o. 60 20 26 15 29 3 p.o. 60 30 40 14 ADJUVANT ARTHRITIS
Heat-killed M~ùbd~lLIiulll LUIY~;LUIII was ground to a find powder and ~"~r ..lAd in heavy mineral oil at 10 mglml. The s ~ o" was injected ~ ~1. m~ u~ y into the base of the tail of male Lewis rats at 0.1 ml per rat. This i~ ,n;~" p,u~e.lu,t: induces an ay,~ arthritis that is apparent at day 10-12 and rapidly worsens. The volumes of the hind paws were l"ea~ d before immunization and on days 12, 15 and 20 after ;~ The baseline paw volume was ~ulllld~ d from the arthritic volumes to yield paw swelling. AK inhibitors were given by daily oral gavage beginning on day 4 after i.. :".~ , using I ~l~d~h,'~ ~~ glycol400 as the vehicle. Control rats received vehicle only. Percent inhibition was calculated based on paw swelling in AK inhibitor treated group cu,.l~Jd,~d to vehicle treated group, and is reported in Table 11.
TABLE 11: PERCENT INHIBITION
Dose # % inh at day 12 % inh at day 15% inh at day 20 ~mglkg~
28 10 29 i 7 22 i 4 28 20 52 i 23 52 + 22 40 i 8 SUBSTITUTE SHEET (RULE 26) CA 02220642 1997-ll-28 W O 96/40705 PCT~US96/10404 FORMALIN PAW
In this assay, injection of formalin, an irritant, into the hindpaw of rats typically evokes a biphasic response of pain-related behaviors. Phase 1 of the response which is brief, lasting appr~xil"~lely O - 5 min post-injection, is followed by 5 a more prolonged phase 2, lasting approximately 10 - 80 min post-inje--tion. Phase 1 behavior is thought to be a direct effect of the irritant on "oc;cep~or~ at the injection site while phase 2 behavior is thought to include a hyperalgesic cc ~ onent merii~ted by sensitization of neuronal elements within the spinal cord. Studies from otherla).o, ~Lories have found the first portion of Phase 2 (sometimes referred to as Phase 10 2a) to be most responsive to pharmacological manipulation.
Rats (male, Si~"onsen) weighing between 100 - 200 9, are used in the present experi"~e"l-~. For screening purposes drugs are administered orally 90 min prior to initiation of formalin test. At designated intervals the animals in groups of 4 are placed individually in a small animal restrainer with the right hindpaw ~ccessihle 15 through a hole in the bottom of the resl, ci"er. The formalin paw assay is initiated by the il ,;ection using a 30G needle of 50 ~11 of a 5% formalin solution in saline into the right plantar surface of each hindpaw. The rat is then immediately placed in a separate plexiglass box and scoring (described below) of the animal's behavior is begun at 1.7 min after formalin in,_otion. The il l~l~l .lal leous behavior of each animal 20 in a group of 4 was observed and assiyl ,ed a score once in each 20 second interval.
This sequence is repeated over a 30 min period. The scorin~ protocol is an adaptation of the method published by Dubuisson and Dennis (Pain 4:161 -174, 1977) which assigns a score from 0 - 3 as fol!ows:

0 - no discernible favoring of injected paw, weight evenly distributed 25 1 - injected paw is favored, rests lightly on floor 2 - injected paw is elevated 3 - injected paw is vigorously licked, bitten or shake Scores are continuously recorded directly onto an Excel spreadsheet.
For comparative examination of drug effects the data is reported in two different 30 ways: 1) the scores are summed for Phase 1 (1.7 - 5 min post-formalin) and for WO 96/40705 PCT~US96/10404 Phase 2110.3 - 30 min post-formalin) and the mean values of the sums are determined from 6 different animals with results ~ u~o;~od as % inhibition culllpdlod to vehicle control. 2) the total number of ~ ' specifically of 'i~ ; ~9 behavior is summed over Phase 2 and mean values determined from 5 6 different animals with results eAI,-essed as % inhibition culllpalrd to vehicle control. Results are shown in Table 12.

IN VIVO ACTIVIT~' OF SELECTED AKIS IN RATS IW THE FORMALIN PA~ MODEL
10Example ~% Inhibiti~n% Inhibiti~nX Inhibition Phase 2.
Phase 1 PhasO 2 1 ' ."
CDm~osite scoreComposite score 27 25.5 47.6 77.5 28 9.2 37A 76.1 29 24.8 35.1 67.3 FORMULATIONS
C~ -d~ of the invention are administered to the affected tissue at the rate of from 0.1 to 200 nmolelminlkg, ~ Or~O~ably from 1 to 50 nmollminlkg. Such rates are easily maintained when soluble c--~I~u~ c are i..l..,~_.,ou;,l~ od as d~,~.,"ed below. When other methods are used (e.g., oral administration), use of time-release ~JIe~Jdldliùns to control the rate of release of the active ingredient may be preferred. These .v~ -- I< are administered in a dose of about 0.01 mglkglday to 20 about 100 mglkglday, ~ Or~OIably from about 0.1 Illylkyl-ldy to about 10 mglkglday.
For the purposes of this invention, the c -- ~po -,~lC of the invention may be administered by a variety of means including orally, parenterally, by i l~ i n spray, topically, or rectally in rw~ i- n, containing cu...~ iulldl non-toxic pl.d...~e.,li~ally aLc~O~-i ' ' carriers, adjuvants and vehicles.
The term pdlOIlLeldl as used herein includes ~uLLuLdll-ouu~ i..L.u~,~..uu~ LId~u~clllar, and illLIddlLOlial 25 injections with a variety of infusion tL' , - hlllddllolidl and illll~.,.o.lduS injection as used herein includes admin;~l,dliu.. through cdLl~-olol:i. F~l-o~o~-od for certain indications are methods of admi..;~L,dLiùn which allow rapid access to the tissue or SU~STITUTE SHEET (RULE 26) W O 96/40705 PCT~US96/10404 organ being lr~dled, such as intravenous injections for the t, e~l" ,~, ll of myoca, dial i"rarcliol ,. When an organ outside a body is being treated, perfusion is prerel, ed.
~ rhc",~celJtic~l col"~,osilions containing the active ingredient may be in any form suitable for the intended method of administration. When used for oral 5 use for example, tablets, troches, lo,e,)ges, aqueous or oil suspensions, dispersible powders or granules, emulsions, hard or soft capsules, syrups or elixirs may be prepared. Compositions intended for oral use may be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such co",posilions may contain one or more agents including those from the grouptO consisting of sweetening agents, flavoring agents, coloring agents and preserving agents, in order to provide a palatable preparation. Tablets containing the active ingredient in admixture with non-toxic pharmaceutically acceptable excipient which are suitable for manufacture of tablets are acceptal,le. These excipients may be, for example, inert diluents, such as calcium carbonate, sodium carbonate, l~ctose, 15 calcium phosphate or sodium phospl ,ale; granulating and disintegrating agents, such as maize starch, or alginic acid; binding agents, such as starch, gelatin or acacia; and lubl ic~ling agents, such as magnesium slearcle, stearic acid or talc. Tablets may be uncoated or may be coated by known techniques including microencars~ tjon to delay disintegration and adsorption in the gastrointestinal tract and thereby provide 20 a sustained action over a longer period. For example, a time delay material such as glyceryl ~I~onoslear~le or glyceryl distearate alone or with a wax may be employed.
Formulations for oral use may be also presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, forexample calcium phosphale or kaolin, or as soft gelatin capsules wherein the active 2~ ingredient is mixed with water or an oil medium, such as peanut oil, liquid ,ua~ ~rrin or olive oil.
Aqueo~ l-s suspensions of the invention contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions.
- Such excipients include a suspending agent, such as sodium carboxymethylcell~ s O, 30 methylcell~ ~lose, hydroxypropylmethylcelluose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum ~c~ci~ and dispersing or wetting agents such as a naturally occurring pl)os,uh~lide (e.g., lecithin), a condensalion product of an alkylene CA 02220642 l997-ll-28 W O 96/40705 PCT~US96/10404 oxide with a fatty acid (e.g., polyoxyethylene slearale~, a condensalion product o ethylene oxide with a long chain ali~l ,alic alcohol (e.g., hepl~de~lhyleneoxyoeta"ol), a cGndel)s~lio" product of ethylene oxide with a partial ester derived from a fatty acid and a hexitol anhydride (e.g., polyoxyethylene sorbitan mono-oleate). The aqueous 5 suspension may also conlai, I one or more preservative such as ethyl of n-propyl p-hydroxybe"~oale, one or more coloring agent, one or more flavoring agent and oneor more sweetening agent, such as sucrose or sacchal i".
Oil suspensio"s may be formulated by suspending the active ingredient in a vegetable oil, such as arachis oil, olive oil, sesame oil or coconut oil, or in a 10 mineral oil such as liquid pd,~r~i". The oral suspensions may contain a thickening agent, such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents, such as those set forth above, and flavoring agents may be added to provide a p~l~t~hle oral preparation. These compositions may be preserved by the z~d~ ol ~ of an antioxidant such as ascorbic acid.
Dispersible powders and granules of th~e invention suitable for preparation of an aqueous suspension by the addition ol water provide the activeingredient in admixture with a dispersing or wetting agent, a suspending agent, and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those ~isclQsed above. Addilional excipients, for example 20 sweetening, flavoring and coloring agents, may also be present.
The plla""aceutical compositions of the invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, such as olive oil or arachis oil, a mineral oil, such as liquid par~rin, or a mixture of these. Suitable emulsifying agents include naturally-occurring gums, such as gum acacia and gum 25 tragacanth, naturally occurring phosphatides, such as soybean lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan mono-oleate, and condensation products of these partial esters with ethylene oxide, such as polyoxyethylene sorbitan mono-oleate. The emulsion may also contain sweetening and flavoring agents.
Syrups and elixirs may be formulated with sweetening agents, such as glycerol, sorbitol or sucrose. Such formulations may also contain a demulcent, apreservative, a flavoring or a coloring agent.

W O 96/40705 PCT~US96/10404 The phal,llaceutical compositions of the invention may be in the form of a sterile il, o~h'~ ,c r~par~Lio, I, such as a sterile injectable ~q~ ~eo~ ~s or oleaginous suspension. This suspension may be formulated according to the known art using those s~ ~it~h'e disper~i, Ig or wetting agents and suspending agents which have been mentioned above. The sterile il ;- ~hlQ preparation may also be a sterile injectable solution or suspension in a non-toxic pal ~nler~lly-acce~lable diluent or solvent, such as a solution in 1,3-butanediol or pre,c,ared as a Iyophylized powder. Among theacceplable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fixed oils may 10 convenlionally be employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid may likewise be used in the preparation of injectables.
The amount of active ingredient that may be combined with the carrier 15 material to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. For example, a time-release formulation intended for oral adminisllaliol) to humans may contain 20 to 1000 ,L~moles of active rl l~l~rial compounded with an appropri~le and convenient amount of carrier l l ,ale, ial which may vary from about 5 to about 95% of the total compositions. It is prafe" ~d 20 that pharmaceutical composition be prepared which provides easily measurable amounts for administration. For example, an aqueous solution intended for intravenous infusion should conlail I from about 0.1 to about 15 ,umoles of the active ingredient per ML of solution so that infusion of a suitable volume at a rate of about 30 mL/hr can occur.
As noted above, formulations of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets each conlai"i"y a predelel I l lil ,ed amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous or non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active30 ingredient may also be administered as a bolus, electuary or paste.
A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by W O 96/40705 PCT~US96/10404 co",pressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder (e.g., povidone, gelatin, hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disi"Ley,~l,L
(e.a., sodium starch glycolate, cross-linked povidone, cross-linked sodium carboxymethyl cell~ ~ose) surface-active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound " ,oi~Lel ,ed with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying 10 pr~po, Lions to provide the desired release profile. Tablets may opLiol ,ally be provided with an enteric coating, to provide release in parts of the gui other than the stomach.
This is particularly advantageous with the compounds of formula (I) as such compounds are s~ ~sceptible to acid hydrolysis.
Formulations suitable for topical administra~ion in the mouth include 15 lozenges comprising the active ingredient in a flavored basis, usually sucrose and acacia or tragacanth; pastilles co" ,,urisi"y the active ingredient in an inert basis such as gelatin and glycerin, or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier. Formulations for rectal administration may be presented as a suppository with a suitable base comprising for example 20 cocoa butter or a salicylate.
Formulations suitable for vaginal administration may be presented as pessaries, tar",uo~ ,s, creams, gels, pastes, foams or spray formulations containing in addition to the ddPN ingredient such carriers as are known in the art to be appropriate.
Formations suitable for pa, ~"aeral administration include aqueous and non-aqueous isotonic sterile injection solutions which may contain anti-oxidants, bu~fers, bacteriostats and solutes which render the formation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions whichmay include suspending agents and thickening agents. The formulations may be presented in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be sorted in a freeze-dried (Iyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Exle",por~neous injection solutions and suspensions may be prep~r~d fromsterile powders, granules and tablets of the kind previously described.
- Preferred unit dosage formulations are those containing a daily dose or unit, daily sub-dose, or an appropriate fraction thereof, of an adenosine kinase 5 inhibitor compound. It will be understood, however, that the specific dose level for any particular patient will depend on a variety of factors including the activity of the specific compound employed; the age, body weight, general health, sex and diet of the individual being treated; the time and route of administration; the rate of excretion; other drugs which have previously been administered; and the severity of 10 the particular ~l~se~se undergoing tnerapy, as is well understood by those skilled in the art.
Capsules comprising adenosine kinase inhibitors suitable for oral administration according to the methods of the present invention may be preparedas follows: (1 ) for a 10,000 c~rsl 'IQ preparation: 1500 9 of adenosine kinase inhibitor 15 is blended with other ingredients (as described above) and filled into capsules which are suitable for ad" lillisll alion depending on dose, from about 4 capsules per day (1 per 6 hours) to about 8 capsules per day (2 capsules per 6 hours), to an adult human.
The compounds of this invention and their preparation and use can be 20 understood further by the, ep, ~se"l~li\/e examples above, which illustrate the various aspects of the invention without limiting its scope.

Claims (68)

WE CLAIM:

1. C-4'-modified pyrrolo[2,3-d] and pyrazolo[3,4-d] pyrimidine nucleoside analogs of Formula 1 wherein:
A and B are both hydrogen, or are each independently alkenyl, the group (CH2)nQ, where n is from 1 to 4 and Q is hydrogen, hydroxy, alkyl, alkoxy, amino, azido, or halogen; or A and B together form a ring of from 3 to 6 carbons, the ring containing 0 to 3 heteroatoms selected from oxygen and nitrogen and optionally substituted by Q as defined above;
D is halogen, aryl, aralkyl, alkyl, alkenyl, alkynyl optionally containing one or more heteroatoms such as nitrogen, oxygen or sulfur, haloalkyl, cyano, or carboxamido;
E is nothing when Y is nitrogen; and is hydrogen, halogen, or alkyl when Y is carbon;
F is alkyl, aryl, aralkyl, halogen, amino, alkylamino, arylamino, aralkylamino, alkoxy, aryloxy, aralkyloxy, alkylthio, arylthio, aralkylthio;
G is hydrogen or halogen;
Y is carbon or nitrogen;

Z1 and Z2 are independently hydrogen, acyl, or taken together form a cyclic carbonate;
and pharmaceutically acceptable salts thereof.

2. A compound of claim 1, where Y is carbon.

3. A compound of claim 1, where Y is nitrogen.

4. A compound of any of claims 1-3 where A and B are the same.

5. A compound of any of claims 1-3 where A and B are the same, but are not both methyl.

6. A compound of any of claims 1-3 where A and B are both hydrogen.

7. A compound of any of claims 1-3 where A and B are different.

8. A compound of any of claims 1-3 where D is halogen, aryl, cyano, or CONRR', where R and R' are independently hydrogen or alkyl.

9. A compound of any of claims 1-3 where F is amino, arylamino,halogen or alkyl.

10. A compound of claim 1, where G and each Z are hydrogen.

11. A compound of claim 2, where E, G, and each Z are hydrogen.

12. A compound of claim 3, where G and each Z are hydrogen.

13. A compound of any of claims 10-12 where A and B are the same.

14. A compound of any of claims 10-12 where A and B are the same, but are not both methyl.

15. A compound of any of claims 10-12 where A, and B are both hydrogen.

16. A compound of any of claims 10-12 where A and B are different.

17. A compound of any of claims 10-12 where D is aryl.

18. A compound of any of claims 10-12 where F is arylamino.

19. A compound of and of claims 1-3 where F is arylamino and D is aryl.

20. A compound of any of claims 10-12 where F is arylamino and D is aryl.

21. A compound of claim 1, where A and B together form a ring of 3 to 6 carbons, the ring containing 0 to 3 heteroatoms selected from oxygen and nitrogen and optionally substituted by hydrogen, hydroxy, alkyl, alkoxy, amino, azido or halogen.

22. A compound of claim 2, where A and B together form a ring of 3 to 6 carbons, the ring containing 0 to 3 heteroatoms selected from oxygen and nitrogen and optionally substituted by hydrogen, hydroxy, alkyl, alkoxy, amino, azido or halogen.

23. A compound of claim 3, where A and B together form a ring of 3 to 6 carbons, a ring containing 0 to 3 heteroatoms selected from oxygen and nitrogen and optionally substituted by hydrogen, hydroxy, alkyl, alkoxy, amino, azido or halogen.

24. A compound according to any of claims 21 and 23, where G and each Z
are hydrogen.

25. A compound according to claim 22, where E, G and each Z are hydrogen.

26. A compound according to any of claims 21-23, where at least one of F is arylamino and D is aryl.

27. A compound according to any of claims 21-23, where F is arylamino and D is aryl.

28. A compound of claim 1, where A and B are independently HOCH2 or CH3, provided both A and B are not CH3.

29. A compound of claim 2, where A and B are independently HOCH2 or CH3 provided both A and B are not CH3.

30. A compound of claim 3, where A and B are independently HOCH2 or CH3 provided both A and B are not CH3.

31. A compound of claim 28, where F is phenylamino optionally substituted at any position by halogen, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, amino, cyano or lower alkyl, and D is phenyl optionally substituted at any position by halogen, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, amino, cyano or lower alkyl.

32. A compound of claim 29, where F is phenylamino optionally substituted at any position by-halogen, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, amino, cyano or lower alkyl, and D is phenyl optionally substituted at any position by halogen, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, amino, cyano or lower alkyl.

33. A compound of claim 30, where F is phenylamino optionally substituted at any position by halogen, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, amino, cyano or lower alkyl, and D is phenyl optionally substituted at any position by halogen, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, amino, cyano or lower alkyl.

34. A compound of claim 21, where F is phenylamino optionally substituted at any position by halogen, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, amino, cyano or lower alkyl, and D is phenyl optionally substituted at any position by halogen, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, amino, cyano or lower alkyl.

35. A compound of claim 22, where F is phenylamino optionally substituted at any position by halogen, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, amino, cyano or lower alkyl, and D is phenyl optionally substituted at any position by halogen, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, amino, cyano or lower alkyl.

36. A compound of claim 23, where F is phenylamino optionally substituted at any position by halogen, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, amino, cyano or lower alkyl, and D is phenyl optionally substituted at any position by halogen, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, amino, cyano or lower alkyl.

37. A compound according to any of claims 28-36 where G and each Z are hydrogen.

38. A compound according to any of claims 28, 29, 31, 32, 34, and 35 where E, G, and each Z are hydrogen.

39. A compound of claim 35, where D is phenyl and F is phenylamino substituted at any position by halogen or alkoxy.

40. A compound of claim 36, where D is phenyl and F is phenylamino substituted at any position by halogen or alkoxy.

41. A compound of any of claims 1-3 where D is halogen or aryl.

42. A compound of any of claims 1-3 where D is aryl.

43. A compound of any of claims 1-3 where F is amino or arylamino.

44. A compound of any of claims 1-3 where F is arylamino.

45. A compound of claim 6 where D is aryl.

46. A compound of claim 6 where F is arylamino.

47. A compound of claim 6 where D is aryl and F is arylamino.

48. A compound of claim 15 where D is aryl.

49. A compound of claim 15 where F is arylamino.

50. A compound of claim 15 where D is aryl and F is arylamino.

51. A compound of claim 6 where D is phenyl optionally substituted at any position by 1 to 2 substituents chosen independently from halogen, lower alkyl, amino, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, (CH2)rT where r is from 0 to 3 and T is an alkyl chain of 0 to 16 carbon atoms containing one or more hetero atoms, N-sulfonylated amino, amidoximo, N-aminoguanidino, amidino, guanidino, acyl guanidino, cyclic derivatives of amidino, guanidino, aminoguanidino, 5 or 6 membered ring containing at least one basic nitrogen and optionally one or more oxygen atoms, and optionally substituted by CONVV' where V and V' are independently an alkyl chain, at least one of which contains one or more basic nitrogen atoms and optionally oxygen atoms, or V and V' together form a 6 membered ring containing at least one basic nitrogen.

52. A compound of claim 6 where F is is phenylamino optionally substituted at any position by 1 to 2 substituents chosen independently from halogen, lower alkyl, amino, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, (CH2)rT where r is from 0 to 3 and T is an alkyl chain of 0 to 16 carbon atoms containing one or more hetero atoms, N-sulfonylated amino, amidoximo, N-aminoguanidino, amidino, guanidino, acyl guanidino, cyclic derivatives of amidino, guanidino, aminoguanidino, 5 or 6membered ring containing at least one basic nitrogen and optionally one or more oxygen atoms, and optionally substituted by CONVV' where V and V' are independently an alkyl chain, at least one of which contains one or more basic nitrogen atoms and optionally oxygen atoms, or V and V' together form a 6 membered ring containing at least one basic nitrogen.

53. A compound of claim 6 where D is phenyl and F is phenylamino and both are optionally and independently substituted at any position by 1 to 2 substituents chosen independently from halogen, lower alkyl, amino, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, (CH2)rT where r is from 0 to 3 and T is an alkyl chain of 0 to 16 carbon atoms containing one or more hetero atoms, N-sulfonylated amino, amidoximo, N-aminoguanidino, amidino, guanidino, acyl guanidino, cyclic derivatives of amidino, guanidino, aminoguanidino, 5 or 6 membered ring containing at least one basic nitrogen and optionally one or more oxygen atoms, and optionally substituted by CONVV' where V and V' are independently an alkyl chain, at least one of which contains one or more basic nitrogen atoms and optionally oxygen atoms, or V and V' together form a 6 membered ring containing at least one basic nitrogen.

54. A compound of claim 15 where D is phenyl optionally substituted any position by 1 to 2 substituents chosen independently from halogen, lower alkyl, amino, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, (CH2)rT where r is from 0 to 3 and T is an alkyl chain of 0 to 16 carbon atoms containing one or more hetero atoms, N-sulfonylated amino, amidoximo, N-aminoguanidino, amidino, guanidino, acyl guanidino, cyclic derivatives of amidino, guanidino, aminoguanidino, 5 or 6membered ring containing at least one basic nitrogen and optionally one or more oxygen atoms, and optionally substituted by CONVV' where V and V' are independently an alkyl chain, at least one of which contains one or more basic nitrogen atoms and optionally oxygen atoms, or V and V' together form a 6 membered ring containing at least one basic nitrogen.

55. A compound of claim 15 where F is is phenylamino optionally substituted at any position by 1 to 2 substituents chosen independently from halogen, lower alkyl, amino, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, (CH2)rT where r is from 0 to 3 and T is an alkyl chain of 0 to 16 carbon atoms containing one or more hetero atoms, N-sulfonylated amino, amidoximo, N-aminoguanidino, amidino, guanidino, acyl guanidino, cyclic derivatives of amidino, guanidino, aminoguanidino, 5 or 6 membered ring containing at least one basic nitrogen and optionally one or more oxygen atoms, and optionally substituted by CONVV' where V and V' are independently an alkyl chain, at least one of which contains one or more basic nitrogen atoms and optionally oxygen atoms, or V and V' together form a 6 membered ring containing at least one basic nitrogen.

56. A compound of claim 15 where D is phenyl and F is phenylamino and both are optionally and independently substituted at any position by 1 to 2 substituents chosen independently from halogen, lower alkyl, amino, hydroxy, alkoxy, perhalo lower alkyl, carboxamido, (CH2)rT where r is from 0 to 3 and T is an alkyl chain of 0 to 16 carbon atoms containing one or more hetero atoms, N-sulfonylated amino, amidoximo, N-aminoguanidino, amidino, guanidino, acyl guanidino, cyclic derivatives of amidino, guanidino, aminoguanidino, 5 or 6 membered ring containing at least one basic nitrogen and optionally one or more oxygen atoms, and optionally substituted by CONVV' where V and V' are independently an alkyl chain, at least one of which contains one or more basic nitrogen atoms and optionally oxygen atoms, or V and V' together form a 6 membered ring containing at least one basic nitrogen.

57. A compound of claim 6 where D is phenyl optionally substituted by halogen.

58. A compound of claim 6 where F is phenylamino optionally substituted by halogen or (CH2)rT where r is from 0 to 3 and T is an alkyl chain of 0 to 16 carbon atoms containing one or more hetero atoms, N-sulfonylated amino, amidoximo, N-aminoguanidino, amidino, guanidino, acyl guanidino, cyclic derivatives of amidino, guanidino, aminoguanidino, 5 or 6 membered ring containing at least one basic nitrogen and optionally one or more oxygen atoms.

59. A compound of claim 6 where D is phenyl optionally substituted by halogen and F is phenylamino optionally substituted by halogen or (CH2),T where r is from 0 to 3 and T is an alkyl chain of 0 to 16 carbon atoms containing one or more hetero atoms, N-sulfonylated amino, amidoximo, N-aminoguanidino, amidino, guanidino, acyl guanidino, cyclic derivatives of amidino, guanidino, aminoguanidino, 5 or 6 membered ring containing at least one basic nitrogen and optionally one or more oxygen atoms.

60. A compound of claim 15 where D is phenyl optionally substituted by halogen.

61. A compound of claim 15 where F is phenylamino optionally substituted by halogen or (CH2)rT where r is from 0 to 3 and T is an alkyl chain of 0 to 16 carbon atoms containing one or more hetero atoms, N-sulfonylated amino, amidoximo, N-aminoguanidino, amidino, guanidino, acyl guanidino, cyclic derivatives of amidino, guanidino, aminoguanidino, 5 or 6 membered ring containing at least one basic nitrogen and optionally one or more oxygen atoms.

62. A compound of claim 15 where D is phenyl optionally substituted by halogen and F is phenylamino optionally substituted by halogen or (CH2)rT where r is from 0 to 3 and T is an alkyl chain of 0 to 16 carbon atoms containing one or more hetero atoms, N-sulfonylated amino, amidoximo, N-aminoguanidino, amidino, guanidino, acyl guanidino, cyclic derivatives of amidino, guanidino, aminoguanidino, 5 or 6 membered ring containing at least one basic nitrogen and optionally one or more oxygen atoms.

63. A compound of claim 6 where D is phenyl, 4-chlorophenyl or 4-fluorophenyl.

64, A compound of claim 15 where D is phenyl, 4-chlorophenyl or 4-fluorophenyl.

65. A compound of claim 58 where r is from 0 to 2 carbon atoms and T is one of amino, alkylamino, dialkylamino, guanidino, acyl guanidino, cyclic amidino, cyclic guanidino, an alicyclic ring containing at least one basic nitrogen and optionally one oxygen atom.

66. A compound of claim 61 where r is from 0 to 2 carbon atomsand T is one of amino, alkylamino, dialkylamino, guanidino, acyl guanidino, cyclic amidino, cyclic guanidino, an alicyclic ring containing at least one basic nitrogen and optionally one oxygen atom.

67. A compound of claim 6 where D is phenyl, 4-chlorophenyl or 4-fluorophenyl and F is phenylamino optionally substituted by halogen or (CH2)rTwhere r is from 0 to 2 carbon atoms and T is one of amino, alkylamino, dialkylamino, guanidino, acyl guanidino, cyclic amidino, cyclic guanidino, an alicyclic ring containing at least one basic nitrogen and optionally one oxygen atom.

68. A compound of claim 15 where D is phenyl, 4-chlorophenyl or 4-fluorophenyl and F is phenylamino optionally substituted by halogen or (CH2)rTwhere