IE83251B1 - AICA riboside analogs - Google Patents

Description

Field of Invention The present invention generally relates to nucleoside analogs, specifically to 5-aminobeta—D—ribofuranosyl- imidazo1e—4—carboxamide ("AICA riboside") analogs. The present invention also relate to the preparation of these compounds and their use in the manufacture of medicaments for use in the treatment of cardiovascular, cerebrovascular, other ischemic disorders, and other diseases which can be regulated by local increase of extracellular adenosine such as inflammatory or thrombotic conditions.

Background of The Invention The present invention is directed at novel compounds which are analogs of AICA riboside. AICA riboside enters cells and is phosphorylated to AICA riboside monophosphate ("ZMP"), a naturally occurring intermediate in purine biosynthesis. AICA riboside is said to increase extracelluar adenosine levels under conditions of net ATP breakdown and, therefore, in light of the cardioprotective and neuroprotective properties of adenosine it may have potential therapeutic uses. However, AICA riboside has a relatively low potency and short half life. Also, we have found that AICA riboside does not cross the blood-brain barrier well and is inefficiently absorbed from the gastrointestinal tract. These characteristics of limited potency, limited oral bioavailability and limited brain penetration decrease its potential for use as a therapeutic agent.

AICA riboside treatment has been reported to have beneficial effects in a number of experimental models of myocardial ischemia. In a dog model, in which pacing induced a profound progressive decline in wall thickening and endocardial blood flow and an increase in ST segment of the EKG, AICA riboside markedly attenuated these changes to maintain contractile deviation intramyocardial function [Young and Mullane, (1991)]. induced by coronary artery occlusion, Am. J. Physio., In another dog model, in which ischemia was AICA riboside was reported to be beneficial by significantly decreasing ischemia-induced arrhythmias and improving blood flow to the ischemic region of the myocardium_ (Gruber et al, Circulation g (5): 1400-1410 (1990)). An effect of AICA riboside to increase regional blood flow and maintain contractile function was also reported in a dog model of coronary embolization in which ischemia was induced by administration of microspheres directly into the coronary circulation (Takashima et (Supplement 4):41 (1990)). A potential consequence of this in blood flow by AICA riboside was said to be a reduction of infarct size (McAllister et al, 303A (1987)).

Treatment with AICA riboside has been reported to have al, Heart favorable consequences in other experimental models of myocardial ‘ischemia. For instance, Mitsos et al (Pharmacology ;;: 121-131 (1985)) reported that AICA riboside improved the recovery of post-ischemic function in the isolated blood—perfused cat heart and Bullough et in press recovery in an isolated buffer-perfused guinea pig heart.

Thus, AICA riboside has ischemia-induced injury to been reported to alleviate the heart in various experimental models.

AICA riboside has also been reported to protect brain _ In a rat model of focal ischemia, AICA riboside treatment hippocampal CA-1 cells, was reported to provide a significant reduction in infarct size. The protective effects of AICA riboside have also been reported in other models of ischemia, including rat.

A number of studies suggest that the beneficial effects of AICA riboside can be ascribed, at least in part, to an increase in local levels of adenosine, which has similar cardioprotective (olafsson et al, Circulation AICA levels is both direct i.e.

Evidence for riboside—induced enhancement of reversal of the anti-ischemic properties of AICA riboside culture models (Gruber et al, i.e. by removal of exogenous adenosine using adenosine deaminase (Young & Mullane, Am.J. Physio., in press (1991)). In hearts subjected to ischemia and reperfusion, cellular damage has been, in part, attributed to plugging of the microvessels by neutrophils. Adenosine has been reported to inhibit neutrophil adhesion to coronary and hence through prevention of neutrophil-dependent tissue injury This is supported by evidence for decreased accumulation of in some models of ischemia and reperfusion. neutrophils in the ischemic region of the heart by AICA riboside (Gruber et al, Circulation gg: 1400-1410 (1990)).

A recognition of the neuroprotective properties of cardioprotective and adenosine have led to attempts to explore the therapeutic use of exogenously' administered adenosine itself. However the short half life of adenosine in blood (<10 secs) necessitates the use of high doses and continuous infusions to maintain levels appropriate for most treatments. Adenosine itself causes hypotension, i.e. reduces blood pressure; it is also a negative chronotrcpic and dromotropic agent, i.e. reduces electrical conduction in the heart, heart rate and respectively. Adenosine would therefore exert marked systemic hemodynamic effects at concentrations that would be required to elicit cardioprotective or neuroprotective properties. These systemic cardiovascular actions are frequently contraindicated in most clinical conditions where adenosine could be useful. In contrast, as a result of its local effects on adenosine levels, AICA riboside administration does not produce such side-effects, even at doses considerably higher than the expected therapeutic H Adenosine receptor agonists have also been studied and effects similar to adenosine have been reported in a (Daly, J. Med. Chem. because most cell types have number of experimental models. (3):l97 (1982).

Again, adenosine receptors, exogenously administered adenosine agonists exhibit profound actions on a variety of tissues and organs, outside of the target organ, thereby limiting their therapeutic potential. other ways of potentially achieving the effect of a high local extracellular level of adenosine have been studied. of adenosine They include: a) interference with the uptake with that specifically block adenosine transport, as described by Paterson et al., in reagents p. 402 (1975); b) adenosine, as described by Carson and Seegmiller in Ihe Journal of Clinical Investigation, Vol. 57, p. 274 (1976); and c) the use of analogs of adenosine constructed to bind prevention of the degradation of to adenosine cell plasma membrane receptors.

There are a repertoire of chemicals that reportedly can inhibit the cellular uptake of adenosine. Some have been reported to do so specifically, and are believed to be essentially competitive inhibitors of adenosine uptake, and others are believed to inhibit nonspecifically. p-nitrobenzylthioinosine appears to be a competitive inhibitor, while dipyridamole and a variety of other chemicals, including colchicine, phenethyalcohol and papaverine appear to inhibit uptake nonspecifically.

U.s. Patent No. 4,115,641 to Fischer et al. is directed to certain ribofuranosyl derivatives which are said to have cardiac and circulatory-dynamic properties.

In particular, Fischer et al. are directed to certain compounds which are said to have intrinsic adenosine—like modes of action as determined by measuring decreased heart rate and blood pressure.

In contrast, AICA riboside and AICA riboside-like lead to at the specific time and location of a pathological event and compounds enhanced adenosine levels thus permit increased adenosine levels to be selectively targeted without the detrimental side effects.

The present invention is directed to AICA riboside analogs which exhibit and, in many cases, improve upon, The novel compounds typically exhibit one or more of the 1) functional more potent the positive biological effects of AICA riboside. following improvements over AICA riboside: adenosine or; 4) increased oral bioavailability and/or brain penetration. benefits at lower doses; 2) regulating actions; 3) increased half-lives Summary of the Invention The present invention is directed to certain new analogs of AICA riboside which exhibit enhanced potency, or metabolism efficacy or improved pharmacokinetics compared to AICA riboside. In particular, the present invention is directed to four series of novel analogs having chemical modification at the following positions: N-4 (Series I), C-2 (Series II), 5'-C (Series III) and 2'- C (Series IV). A number of compounds in these preferred series provide improved functional recovery of post ischemic function at lower _concentrations than AICA riboside. The beneficial effects of these compounds result, at least in part, from their ability to increase extracellular adenosine levels more effectively than AICA riboside. Moreover, some of these compounds are inhibitors of adenosine transport and individual adenosine-regulating enzymes.

The AICA riboside analogs of this invention are useful in treating a variety of clinical situations where increased extracellular levels of adenosine would be bene- ficial. Accordingly, the compounds may be used in the conditions as heart attack, treatment of such PTCA, ischemia-related diseases, prophylactic and affirmative angina, cardioplegia, stroke and other as well as seizures and inflammatory disorders. This invention is also directed to pharmacological compositions comprising an effective amount of the analog of the present invention and a pharmaceutically acceptable carrier.

Qefinitions As used herein, the following terms have the following meanings, unless expressly stated to the contrary.

The term "hydrocarbyl" refers to an organic radical comprised of primarily carbon and hydrogen and includes alkyl, groups including aryl and aralkyl groups and groups which have a mixture of saturated and unsaturated bonds, alkenyl and alkynyl groups, as well as aromatic ali- cyclic (carbocyclic or cycloalkyl) groups or such groups substituted with aryl (aromatic) groups or combinations thereof and may refer to straight-chain, branched-chain or cyclic structures or to radicals having a combination thereof.

The term "alkyl" refers to saturated aliphatic groups, including straight, branched and carbocyclic groups. The term "lower alkyl" refers to both straight- and branched-chain alkyl groups having a total of from 1 to 6 carbon atoms and includes primary, secondary and tertiary alkyl groups. Typical lower alkyls include, for ethyl, n—propy1, isopropyl, n-butyl, isobutyl, t-butyl, n-pentyl, n-hexyl, and the like.

The term "aryl" refers to aromatic groups having from example, methyl, about 6 to 14 carbon atoms and includes cyclic aromatic systems such as phenyl and naphthyl.

The term "aralkyl" refers to an alkyl group of about 1 to 4 carbon atoms substituted with an aryl group of from 6 to 10 carbon atoms and includes, for example, benzyl, p- chlorobenzyl, p-methylbenzyl and 2-phenylethyl.

The term "alkenyl" refers to unsaturated alkyl groups having at least one double bond [e.g. CH3CH=CH(CH2)2-] and includes both straight and branched-chain alkenyl groups.

The term refers to unsaturated groups having at least one triple bond [e.g. CH3CEC(CH2)2-] and "alkynyl" includes both straight chain and branched-chain groups.

The term chlorine, bromine and iodine. "halo" or "halogen" refers to fluorine, The term "acyl" refers to the group R'C- wherein R‘ is hydrocarbyl.

The term "alkylene" refers to straight, branched- chain and carbocyclic alkylene groups which are biradicals, and includes, for example, groups such as CH3 ethylene, propylene, 2-methylpropylene (e.g. -CHZCHCH2-), CH3 ,6—n-hexylene, 3-methylpentylene (e.g. -CH2CH2CHCH2CH2-), 1,4-cyclohexylene, and the like.

-CNR"2 wherein each R" is independently hydrogen or hydrocarbyl, or to compounds having at least one such group.

O The term "carboxamide" refers to the group -CNR"2 wherein each R" is independently hydrogen or hydro- carbyl. The term "unsubstituted carboxamide" refers to O the group -CNH2.

O The term "acylamino" refers to the group -NHCR' wherein R’ is hydrocarbyl. The term "lower acylamino" refers to acylamino groups wherein R‘ is alkyl of 1 to 6 carbon atoms. ' The term "carbonate ester“. refers to the group -OCOR' wherein R‘ is hydrocarbyl or to compounds having at least one such group. 0 refers to the group -OCR‘ wherein R‘ is hydrocarbyl or to compounds having at least The term "acyl ester" one such group.

The term "phosphate ester" refers to the group O -OP-OR" wherein R" is independently hydrogen or hydrocarbyl and/or to compounds having at least one such group, and includes salts thereof.

The term "mixed ester" refers to compounds having at least one carbonate ester group and at least one acyl ester group or to compounds having combinations of different acyl ester or carbonate ester groups.

The term "carboxylic acid ester" or "carboxy ester" refers to the group -COR‘ wherein R‘ is hydrocarbyl or to compounds having at least one such group.

The term "carbocyclic AICA riboside" refers to an analog of AICA riboside wherein the oxygen atom in the ribosyl ring has been replaced by a methylene (-CH2-).

The term "hydrocarbyloxy" refers to the group R'0- wherein R‘ is hydrocarbyl.

The term "alkoxy" refers to the group R'0- wherein R‘ is alkyl.

The term "hydrocarbylthio" refers to the group having the formula R'S- wherein R‘ is hydrocarbyl.

The term NHR‘ or -NR‘: "hydrocarbylamino" refers to the groups- where R‘ is an independently selected hydrocarbyl group.

The term "hydrocarbylimidate" refers to the group NH -C—0R' wherein R" is hydrocarbyl.

The term "carboxamideoxime" refers to the group The term NOR‘ -C—NH2 wherein R‘ is hydrocarbyl. "hydrocarbyloxyamidine refers to the group The term "hydrocarbyloxycarbonyl refers to the group 0 R'-o-C- wherein R‘ is hydrocarbyl.

The term "hydrocarbyloxycarboxy" refers to the group 0 II R‘-O-C-O-wherein R‘ is hydrocarbyl.

The term wherein R‘ is hydrocarbyl. "thioester" refers to the group -C-S-R‘ Brief Description of the Drawings Fig. 1 depicts a comparison of the dose-dependent effects of AICA riboside (Compound No. 1 of Tables XII and XIII’ (1-110)) (Series I) substituted AICA riboside (1-186)) adenosine levels in a rat heart ischemia model. and an N-4 analog (Compound No. 10 on tissue Fig. 2 depicts a comparison of the effect of AICA riboside (Compound No. 1 and a series of 2'-(Series IV) substituted AICA riboside analogs .20 (1-188), 34(1-250) and 32 (1-262)) on utilization of adenosine (together with inosine and hypoxanthine) cell culture model.

(Compound Nos. in a Fig. 3 the substituted AICA riboside analogs of N-4 (Series I) (Compound Nos. 10(1- ) and 11(1-226)) in a gerbil brain ischemia model.

Fig. 4 depicts inhibition of adenosine transport in depicts effects WI—L2 lymphoblasts after 1 minute preincubation with Compound No. 53 (1-468) at the noted concentrations.

‘Hereinafter "Compound No. _" refers to compounds listed in Tables XII amdxm. -11..

Fig. 5 depicts inhibition of adenosine transport in WI-L2 lymphoblasts after 1 hour preincubation with Compound No. 53 (1-468) at the noted concentrations.

Detailed Description of the Invention Preferred AICA Riboside Analoqg The present invention provides analogs of AICA riboside which are compounds of the formula I n,o on, or a pharmaceutically acceptable salt thereof wherein X is or ‘CHf‘; R, is hydrogen, amino, hydrocarbylamino, acylamino, or dihydrocarbylaminoalkyleneimino; Rzis hydrogen, cyano, hydrocarbylimidate, carboxamideoxime, hydrocarbyloxyamidine, carboxamide, or carboxylic acid or an amide, ester, thioester or salt thereof; R3 is hydrogen, hydrocarbyl, amino, hydrocarbylamino, halogen, hydroxy (including tautomeric 2-imidazolone), hydrocarbyloxy, sulfhydryl (including tautomeric 2-imidazolthione), or hydrocarbylthio; R4 and R5 are independently hydrogen, hydrocarbyl, acyl or hydrocarbyloxycarbonyl; R6 is hydrogen, hdyrocarbyl,-halogen, hydroxy, hydrocarbyloxy, sulfhydryl, hydrocarbylthio, sulfamyloxy, amino, hydrocarbylamino, azido, acyloxy or hydrocarbyloxycarboxy or phosphate ester group or salts thereof; provided that when R, is amino, R2 is unsubstituted carboxamide, R3 is hydrogen; R4 and R5 are hydrogen, acyl, or hydrocarboxycarbonyl; then Réis not hydroxy, acyloxy or hydrocarbyloxycarboxy, for use in a method of reducing or preventing tissue damage associated with undesired decreased blood flow.

Since compounds of the above formula wherein R,is hydroxy or sulfhydryl may exist in their isomeric (tautomeric) imidazoleone and imidazolethione forms, these isomers are intended to be included in the ambit of Formula I.

Preferred compounds include those wherein (i) R1 is amino, R2 is carboxamide wherein one of the amide hydrogens is replaced by a hydrocarbyl group, more preferably an aralkyl group (such hydrocarbyl or aralkyl group is substituted, those set forth below); R3 is hydrogen, R‘ hydrogen or hydrocarbyloxycarbonyl, more preferably and R6 optionally suitable suhstituents include and R5 are is hydroxy or amino (Series I); (ii) R1 is amino, R2 is cartoxamide, R3 is halogen or sulfhydryl, R4 is hydrogen, R5 is hydrogen and R6 is hydroxy (Series II); (iii) R1 is amino, R2 is carboxamide; R3, R‘ and R5 are hydrogen and R6 (Series III) (iv) R1 carboxamide, R3 is hydrogen, R‘ is alkyl, R5 is hydrogen is amino and is amino, R2 is and R6 is hydroxy (Series IV).

In particular, in view of their demonstration of activity in various experimental models, compounds include Compound Nos. 10, 23, 25,'29, 47, 52, 53 (Series I), 27, 43 (Series II), 21, 66 (Series III) and , 34 (GP250) and 32 (GP262) (Series IV) of Tables XII and XIII. preferred _ cu:-; R Preferred ACIA Riboside Analogs one preferred group of compounds of formula I include certain AICA riboside analogs wherein X is or — is amino, hydrocarbylamino or dihydrocarbylaminoalkyleneimino, Rzis carboxamide wherein one of the amide hydrogens (attached to the nitrogen atom) is optionally replaced by alkyl, cycloalkyl, or aryl or aralkyl, optionally substituted with 1 to 3 substituents independently selected from halogen, alkyl, aryl, nitro, sulfhydryl, hydrocarbylthio, amino, hydrocarbylamino, 'hydroxy, hydrocarbyloxy, trifluoromethyl, or sulfonamide; R2 is carboxamide wherein both amide hydrogens are replaced by alkyl or together by an alkylene or aralkylene group to form a ring; or R2 is -c(o)—s-R7 wherein R7 is alkyl, cycloalkyl, aryl or aralkyl optionally substituted with 1 to 3 substituents independently selected from halogen, alkyl, aryl, nitro, amino, hydrocarbylamino, sulfhydryl, hydrocarbylthio, hydroxy, hydrocarbyloxy, trifluoromethyl or sulfonamide; R3 is hydrocarbylamino, halogen, hydroxy hydrocarbyl, hydrogen, amino, imidazolone), -imidazolthione) or (including tautomeric sulfhydryl hydrocarbylthio; R4 (including tautomeric and R5 are independently hydrogen, (of 1 to about 18 carbon atoms), acyl or hydroxy, hydrogen, hydrocarbyloxy, sulfhydryl, amino, hydrocarbylamino, hydrocarbyl hydrocarbyloxycarbonyl; and R6 is hydrocarbyl, halogen, hydrocarbylthio, sulfamyloxy, azido, acyloxy, hydrocarbyloxycarboxy or phosphate ester or salt thereof; provided that when -X- is or -CH2-, R1 hydrogen, R4 and R5 independently are hydrogen, acyl or hydrocarbyloxycarbonyl, then R6 is not hydrogen, hydroxy, is amino, R2 is unsubstituted carboxamide, R3 is acyloxy or hydrocarbyloxycarboxy or when R‘ and R5 are both hydrogen, then R6 is not a phosphate ester; when X is oxygen, R1 is amino, R2 is unsubstituted carboxamide, R3 is sulfhydryl, and R‘ and R5 are both hydrogen, then R‘ is not acetoxy; when X is oxygen, R1 is amino, R2 is unsubstituted carboxamide and R3 is chloro, bromo, amino or methoxy, andi R4 and R5 both hydrogen, then R‘ is not hydroxy or when R‘ and Rs are both acetyl, then R‘ is not acetoxy; and provided further that when X is oxygen, R1 is amino, R2 is benzylcarboxamide or p-iodophenylcarboxamide, R3 is hydrogen, then R‘ and R5 are not both hydrogen and R‘ is not hydroxy; or when R: is p—iodophenylcarboxamide, then R‘ and R5 are not both acetyl and R6 is not acetoxy.

Preferred compounds include those whereiJ1R1 is amino, R2 is carboxamide substituted with an aralkyl group, more preferably a benzyl group, having from 1 to 3 ring substitutions as described above, or cycloalkyl. In view of their . preferred compounds include Compound No.5 23, 25, 29, 47, 52 and 53.

One example of an especially preferred compound is a oxygen, R1 chlorobenzylcarboxamide, R3, R‘ and R5 are hydrogen and R6 activity in various experimental models, compound where X is is amino, R5 is p- is amino and salts thereof. one particularly preferred salt is the hydrochloride salt.

Preparation of Preferred ACIA Riboside Analogs The substituted imidazole analogs can be synthesized by well chemical reactions as demonstrated in the examples which follow. (i) prepared from 4-methy1—5-nitro-1H-imidazole by the route known In general, compounds of formula can be described by Baker et al (Baker D., J. Org. Chem. g1:34S7 (1982)) v to carboxylic acid, prepare 1-benzylnitro-1H-imidazole followed by the additional step ‘of reducing the nitro group to give the desired amino group at R1. A the elegant synthesis of AICA riboside reported by Ferris et al. (Ferris, J.P., J. Org.

Alternatively, appropriately protected riboside and diaminomaleonitrile. (1985), -aminoimidazoles starting This route also allows for the introduction of the desired R3 alkyl, hydrocarbyl and aryl groups by selection of the appropriate ortho ester in the cyclization reaction of the the other substituents can be introduced by the methods described by Miyoshi et al. (Miyoshi T., Chem. Pharm. Bull. z4(2):2089 (1976) for the preparation of 2-bromo and 5-aminothio- maleonitrile to imidazole. desired R3 —amino, and 2—hydroxy (as substituted Compounds where the desired R1 tautomeric 2—imidazolones)> 5-amino substituent is acylamino can be prepared by acylation of the corresponding appropriately protected R1 amino compound with the desired acyl anhydride followed by deacylation with ammonia or sodium methoxide. Compounds where R1 is alkylamino or arylamino can be prepared by reductive alkylation of the corresponding appropriately protected R1 amino compound with the desired hydrocarbyl amine as described by Sato et al. (Chem. Pharm. Bull. ;1:1604 (1989)).

Preparation of compounds where R6 "is acyloxy or hydrocarbyloxycarboxy can be prepared selectively by reaction of the appropriate hydrocarbyl acid anhydride or with the 2',3'-C isopropylidene protected riboside followed by removal of supra). Compounds according to formula (I) where R6 is sulfhydryl, prepared from the 5'-deoxy-5'-iodo—2',3‘-isopropylidene hydrocarbylthio or hydrocarbylamino can be ;1:6189 (1971)) for nucleoside phosphates.

Compounds according to formula (I) Utility The AICA riboside analog compounds of this invention will be particularly useful in the reduction of injury during or prevention of ischemia-related events i.e. conditions that arise because of restriction. of blood supply. This attack, infarction, a situation that follows from obstruction of includes heart or myocardial one or more of the coronary arteries supplying blood to the heart muscle, or myocardium, and which, if prolonged, leads to irreversible tissue damage. like AICA riboside, adenosine, Compounds which, lead to increased local levels of and thereby increasing blood flow to the ischemia myocardium, will ameliorate this tissue damage. for a heart attack is thrombolytic therapy, which involves administering a clot one current treatment dissolving agent such as streptokinase or tissue plasminogen activator factor (tPA). However, these drugs must be used within a few hours (1-3) of the heart attack and their effectiveness decreases dramatically with longer delay. The compounds of the present invention, which can be administered prophy£.ctically (i.e, before the event) to achieve a benefit, would therefore clearly be useful.

Angina pectoris is a condition in which the blood supply is sufficient to meet the normal needs of the heart but insufficient when the needs of the heart increase (e.g. becomes more limited (e.g. during coronary artery spasm). during exercise), and/or when the blood supply Patients with angina pectoris or with related conditions such as transient ischemic episodes or silent ischemia could similarly benefit from such an adenosinergic intervention.

In advanced coronary artery disease or persistent chest pain at rest, a number of clinical procedures are currently used to improve blood supply to the heart.

These include percutaneous transluminal coronary angioplasty (PTCA), also known as angioplasty; percutaneous transluminal directional coronary atherectomy, laser atherectomy, intravascular stents and coronary artery bypass graft surgery. The compounds of this invention will also be useful as adjunctive therapies to these techniques.

Another factor lending to cardiovascular problems is which lead to deficiencies in the ability of the heart to supply blood. abnormal heart rhythm, or arrhythmias, The ability of these compounds, like AICA riboside, to reduce arrhythmias will also make them useful in suppressing this condition.

Stroke and central nervous system (CNS) trauma conditions resulting from reduced blood supply to the CNS and is thus amenable to an intervention that provides increased levels of adenosine to the compromised tissue to facilitate tissue survival. other indications ameliorated by agents effecting regional blood flow include organ transplantation, skin flap grafting in reconstructive surgery, peripheral vascular disease, endotoxemia, hemorrhagic shock, pulmonary edema, (thermal pulmonary hypertension, pulmonary injury) or microembolization, injury secondary to burns septicemia, impotence, glomerulonephritis or progressive glomerulosclerosis, vasculitis cardiomyopathies and cardiopulmonary arrest. artherosclerosis, myocarditis, and function. eet al. suggest additional Adenosine has been reported to be an endogenous modulator of inflammation by virtue of its effects on stimulated granulocyte function (Cronstein et al., Q; Clin. _7_8:760-770 (l986)) lymphocyte and platelet function. invention will therefore be useful in conditions in which Invest. and on macrophage, The compounds of this inflammatory processes are prevalent such as arthritis, osteoarthritis, autoimmune (ARDS), necrotizing enterocolitis, disease, adult respiratory distress syndrome inflammatory bowel disease, chronic obstructive pulmonary disease (COPD) and other inflammatory disorders.

Adenosine has been proposed to serve as a natural (Lee et al., Brain Res. 32;:1650-1654 (1984); Dunwiddie, 1nt. Rev. Neugobiol. ;1:63-139 (1985)). anticonvulsant Agents that enhance adenosine levels will therefore be useful for the treatment of seizure disorders. In a recent study, Marangos et al., gpilepsia ;;:239-246 (1990) reported that AICA riboside was an inhibitor of seizures in an experimental animal model.

AICA riboside analogs will also be useful in the treatment of patients who might have chronic low adenosine levels or who might benefit from enhanced adenosine, such as those suffering from autism, cerebral palsy, insomnia, anxiety, or other neuropsychiatric symptoms or those suffering from irritable bowel syndrome. Indeed, a number of studies (Komhuber and Fischer Neurosci. Lett. ;g:32 (1982); Kim et al. Eur. Neurol. g;:367 (1983)) have linked excitatory amino acids with the pathophysiology of schizophrenia. ' The compounds of this invention may also be useful in treating other conditions in which AICA riboside itself has beneficial effects. For instance, since AICA riboside has been reported to have anti-allergic actions in a guinea pig model of bronchospasm induced by antigen sensitization (Bergren et al., submitted to J. of Allergy and Clinical Immunology (1990)), AICA riboside analogs may have therapeutic benefit in the treatment of asthma, hayfever or allergic diseases.

The AICA riboside analogs of the present invention are therefore useful in the treatment of a variety of clinical situations extracellular where increasing adenosine levels and in some cases, at the same time, providing free radical scavenging and/or antioxidant activity are beneficial.

Compounds of the invention are administered to the at the rate of from 0.01 to 3.0 preferably from 0.1 to 1.0 umol/min/kg. such rates are easily maintained when these compounds are intravenously administered as discussed below. When other methods are used (e.g., oral administration), use of time- release preparations to control the rate of release of the affected tissue pmole/min/kg, active ingredient may be preferred. These compounds are administered in a dose of about 0.01 mg/kg/day to about 200 mg/kg/day, about 100 mg/kg/day.

The compounds of the invention may be administered by a variety of means spray, containing preferably from about 0.5 mg/kg/day to including orally, parenterally, by inhalation topically, or rectally in formulations conventional non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles. The term parenteral as used herein includes subcutaneous, intravenous, intramuscular, and intraarterial injections with a variety of infusion techniques. Intraarterial and intravenous injection as used herein includes administration through catheters. Preferred for certain indications are methods of administration which allow rapid access to the tissue or organ being treated, such as intravenous injections for the treatment of myocardial infarction. outside a body is being treated, perfusion is preferred.

Pharmaceutical compositions containing the active ingredient may be in any form suitable for the intended method of administration. when used for oral use for tablets, suspensions, dispersible powders or granules, emulsions, example, troches, lozenges, aqueous or oil 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 compositions may contain one or more agents including those from the group consisting of sweetening agents, and preserving agents, Tablets flavoring agents, coloring agents in order to provide a palatable preparation. containing the active ingredient in admixture with non- toxic pharmaceutically acceptable excipient which are suitable for manufacture of tablets are acceptable. These excipients may be, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium when an organ phosphate or sodium phosphate; granulating and disintegrating agents, such as maize starch, or alginic acid; binding agents, such as starch, gelatin or acacia; and lubricating agents, such as magnesium stearate, stearic acid or talc. Tablets may be uncoated or may be coated by known techniques including microencapsulation to in then intestinal tract and thereby provide a sustained action delay disintegration and adsorption gastro- over a longer period. For example, a time delay material such as glyceryl monostearate 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, for example calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water Aor an oil medium, such as peanut oil, liquid paraffin or olive oil.

Aqueous 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 carboxymethyl- cellulose, methylcellulose, hydroxypropylmethylcelluose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia, and dispersing or wetting agents such as a (e.g., lecithin), a condensation product of an alkylene oxide with a fatty acid (e.g., product of ethylene oxide with a long chain aliphatic naturally occurring phosphatide polyoxyethylene stearate), a condensation alcohol (e.g., heptadeaethyleneoxycetanol), a condensation product of ethylene oxide with a partial ester derived fatty hexitol (e.g., polyoxyethylene sorbitan mono-oleate). aqueous from a acid and a anhydride The suspension may also contain one or more preservative such as ethyl of n-propyl p-hydroxybenzoate, coloring agent, one or more one or more flavoring agent and one or more sweetening agent, such as sucrose or saccharin.

Oil suspensions 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 mineral oil such as liquid paraffin. 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 palable oral preparation. These compositions may be preserved by the addition of an antioxidant such as ascorbic acid.

Dispersible powders and granules of the invention suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient 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 disclosed above. Additional excipients, for example sweetening, flavoring and coloring agents, may also be present.

The pharmaceutical 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 paraffin, or a mixture of these. Suitable emulsifying agents include naturally- occurring gums, such as gum acacia and gum 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 Such formulations may also contain a demulcent, a preservative, agents, such as glycerol, sorbitol or sucrose. a flavoring or a coloring agent.

The pharmaceutical compositions of the invention may be in the form of a sterile injectable preparation, such as a sterile injectable aqueous or oleaginous suspension, This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and The sterile injectable preparation may also be a sterile suspending agents which have been mentioned above. injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, such as a solution in 1,3-butanediol or prepared as a lyophylized powder. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fixed oils may conventionally 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 material to produce a single dosage form will the host treated nd the particular mode of administration. a time- vary depending upon For example, release formulation intended for oral administration to humans may contain 20 to 200 pmoles of active material compounded with an appropriate and convenient amount of carrier material which may vary from about 5 to about 95% of the‘ total pharmaceutical composition be prepared which provides compositions. It is preferred that easily measurable amounts for administration. Fc: example, an aqueous solution intended for intraveno infusion should contain from about 20 to about 50 umoles of the active ingredient per milliliter of solution in order that infusion of a suitable volume at a rate of about 30 ml/hr can occur.

It will be understood, that the specific dose level for any particular patient will depend on a however, 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; other and the severity of the particular disease undergoing therapy, as the rate of excretion; drugs which have previously been administered; ‘is well understood by those skilled in the art.

Examples of use of the method of the invention It will be understood that these and that the method of the invention is not limited solely to these examples. includes the following. examples are exemplary The method may be used following thrombolysis for coronary occlusion. The compound would be given as a sterile injectable preparation with water or isotonic sodium chloride as the solvent. The solution can be administered intravenously or directly into the coronary artery at the time of left heart'catheterization or into a carotid artery. The rate of administration could vary from 0.2 to 1 pmole/min/kg with, for example, an infusion volume of 30 ml/hr. be about 96 hours.

Duration of therapy would typically Angina and early myocardial infarcts can be treated by intravenous administration using a sterile injectable preparation using the rates discussed above.

Compounds of the invention can also be administered to patients intravenously during cardiac bypass surgery or to other surgical patients at risk for a myocardial infarct. The compound can be added directly to the solution administered by the membrane oxygenation, or to the cardiac preservation solution, at the rates discussed above.

Organs can be preserved using the method of the the containing a compound of the invention. with a solution The dosage administered would vary with the rate of perfusion of the invention by perfusing organ organ, as is well understood to those skilled in the art.

This method is particularly applicable to organs and tissues used in organ transplantation.

Qescription of Preferred Embodiments We have identified a number of analogs of AICA riboside that function in experimental models of ischemia. improve the recovery of post-ischemic Table I, the benefit that results from treatment with the preferred analogs is at least equal to AICA riboside 11, 40 (series I), and 19 (Series III)), and in many examples achieved at lower concentrations than AICA riboside (e.g. Compound Nos. 10, 23, 25, 29, 47, 52, 53 (Series I), 27 (Series II), 21, and 66 (Series III)).

In functional assays, which specifically compounds for their ability to increase extracellular adenosine levels, (Compounds Nos. evaluate many of these preferred analogs show The results of evaluating the compounds for their ability‘to markedly enhanced potency compared to AICA riboside. inhibit stimulated contraction in the isolated ileum, an adenosine—mediated functional response, showed that these compounds in each of the preferred series were more effective than AICA riboside (Table II). In addition, the N-4 substituted AICA riboside analogs (Series I) enhanced both tissue adenosine levels in ischemic rat hearts (Table III) inhibited adenosine utilization in coronary endothelial cells (Table IV) degree than AICA riboside. this preferred series (I) also bind with greater affinity and to a significantly greater A number of compounds from to the NBTI-specific adenosine transport site (Table V).

These data suggest that the improved functional benefit of this preferred analog series compared to AICA riboside arises, at least in part, from their ability to increase extracellular adenosine levels and that this ability may be accounted for by inhibition of adenosine transport. (see Table V The C-2 substituted AICA riboside analogs (Series II) also appear to augment exemplified by the effects of Compound No. 13 on adenosine production in cell culture (Table VI) . inhibitors of the adenosine metabolizing enzyme, adenosine and Figures 4 and 5). adenosine release as Moreover, certain of these compounds are As shown in, kinase (see Table VII). The 2'-C substituted AICA riboside analogs (Series IV) profoundly modulate adenosine utilization in a cell culture model (Figure 2). In this "preferred series (IV), each of the test compounds is also an effective inhibitor of adenosine deaminase, another important enzyme (Table VII).

Thus, these compounds increase extracellular adenosine levels more effectively than AICA riboside and this can be explained by enhanced inhibition of adenosine deaminase.

AICA riboside analogs have also been evaluated for their effects on platelet function. certain compounds inhibit platelet aggregation in human whole blood. of the test compounds is enhanced in the presence of a adenosine-metabolizing As shown in Table IX, Inhibition of platelet aggregation by many Adenosine has been reported to be a potent antiplatelet agent, but with a short half life in blood. Accordingly, the inhibition of platelet aggregation observed in the presence of these AICA riboside due to the regulating activity of these compounds.

Certain preferred AICA riboside analogs (Compound No. 53 (1-468), Compound No. 21 (1-227)) are also orally bioavailable in the dog (see Table X). Furthermore, treatment with the AICA riboside analog Compound No. 53 (1-468), provided functional benefits in a canine model of (see Table XI). In addition to their cardiovascular benefits, certain AICA riboside analogs (Compound Nos. 10 (1-186) and 11 (I-226) (Series I)) also have demonstrated protective effects in a gerbil model of brain ischemia (Figure 3). non-inhibitory concentration of adenosine. analogs may be adenosine stable angina To assist in understanding the invention, the results of a series of experiments are presented that demonstrate the benefit of these preferred analogs in models of ischemia and, moreover, provide a rationale for these analogs exhibiting enhanced potency compared to AICA riboside. Also presented are a series of Examples which exemplify the synthesis of these compounds. These should specifically limiting the invention and such variations of examples not, of course, be construed as the invention, now known or later developed, which would be within the purview of one skilled in the art are considered to fall within the scope of the invention as described herein and hereinafter claimed.

Examples Example ; Improved Functional Recovery in Isolated Hearts The ability of a number of the preferred AICA riboside analogs to improve recovery of post—ischemic cardiac function was examined in an isolated rat heart model.

Isolated rat hearts were cannulated via the ascending aorta and attached to a perfusion apparatus according to the method of Langendorff. The hearts were perfused at a constant pressure of 100 cm of H20 with a modified Krebs- Henseleit buffer (pH 7.4) at 37°C. function, left ventricular developed pressure (LVDP) was continuously monitored.

As a measure of heart Following equilibration of the hearts for a period of 30 min., the hearts were subjected to reduced flow i.e. ischemia, by reducing the pressure to cm of H20 for 30 min. Flow was then restored by returning the pressure to its original level (100 cm of H20) for a further 30 min. Each of the AICA riboside A with AICA itself, comparison, was added to the perfusion buffer to a final concentration of 5 uM or 20 pM.

Table I. analogs together riboside for The results are shown in Hooo.

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TABLE II Series Comgound No . (1-110) 23 24 29 39 41 42 44 45 47 53 27 43 21 26 32 (1-225) (1—232) (1-343) (1-354) (1-360) (1—349) (1-355) (1-390) (1—39s) (1—431) (1-434) (1-438) (1-450) (1-468) (1-388) (1—395) (1-432) (1—227) (1-332) (1-262) _Ig5.,__Lw_x >1000 200 60 60 500 1100 70 500 200 800 200 133 Example 3 Effect of AICA Riboside Analogs (Series ) in thg_ Rat Heart Ischemia Model Series I(N-4) substituted AICA riboside analogs were tested for their ability to enhance tissue adenosine levels in ischemic rat hearts.

Male rats were injected intraperitonealy with either the AICA riboside analog, AICA riboside or saline, as a control. After 60 minutes, the hearts were excised and incubated at 37°C for a further 60 minutes. Tissue extracts were prepared and analyzed for adenosine by high performance liquid chromatography (HPLC). The ability of this preferred series of AICA riboside analogs to increase tissue adenosine levels compared to AICA riboside is shown in Table III. A more detailed comparison of the dose- dependent effects on tissue adenosine levels of a selected AICA riboside analog in this preferred series (Compound No. 10) compared to AICA riboside shown in Figure 1.

(Compound No. 1) is TABLE III Tissue Adenosine Levels Compound No.

(% Increase vs. saline) 1 (1-110) 30 (1-186) (Expt 1) 79 (1-186) (Expt 2) 68 11 (1-226) (£xpt 1) 53 ll (1-226) (Expt 2) 45 12 (1-232) 29 (1-207) 34 Exam e 4 nhib't'o o de osi e t’ na o s se ‘es Ce Cu tu e Effects of Series I (N-4) substituted AICA riboside analogs on adenosine utilization were compared using 'zatio b ICA i'bos'd coronary endothelial cells in culture. In this assay, endothelial cells were incubated with 5 pH or 50 pH of the test compound together with 1 uM [3H] adenosine for 15 minutes. Inhibition of adenosine utilization was determined by measuring the concentration of extracellular adenosine by scintillation counting following_separation by thin layer chromatography (TLC). The results of this evaluation are shown in Table IV.

Known compound Example 5 gffiect of A;CA Riboside Analogs (Series ) in [afi]-EBEL fiigdigg Assay The ability of selected Series I (N-4) substituted AICA riboside analogs to effect the binding of [3H]- (NBTI) Increasing concentrations of the test compounds nitrobenzyl-thioinosine to cell membranes was compared. were incubated for 30 minutes with 0.5 mg neuronal membrane protein together with 0.5 nM [3H]-NBTI in a Tris buffer (pH 7.4) The assays were quenched and membranes collected by rapid- filtration.

Filters were then solubilized and radioactivity determined at room temperature. by scintillation counting. The concentration of each test compound which resulted in 50% displacement of bound [3H]- NBTI, the ED5o's, are detailed in Table V. ggzigg Comgound No. (1-110) 29 28 23 39 44 45 46 47 48 49 50 51 52 53 55 56 57 58 59 60 61 64 27 43 54 (1—1ae) (1—3s4) (1-ass) (1-349) (1-aeo) (1-348) (1-343) (1-388) (1-390) (1-434) (1-438) (1-445) (1-450) (1—4s2) (1—453) (1-459) (1—455) (1-457) (1—4es) (1-434) (1—4e7) (1-488) (1-489) (1-506) (1-sos) (1-509) (1-519) (1-395) (1-432) (1-433) TABLE V :r:_I25.,.uz_m >1000 350 300 190 100 72 3 225 600 100 90 8 0.5 22 7 28 16 80 about 100 60 32 80 80 2 17 32 48 344 71 Example 5a lnhibition of Adenosine Eransport in W;-L2 Lymphoblasts Inhibition of adenosine transport in WI-L2 lymphoblasts in the presence of one of the AICA riboside analogs of the present invention was determined according to the following procedure.

A 200 pl aliquot of WI-L2 lymphoblast cell suspension (0.5 X 106) was layered on top of 100 pl of a silicone oil: mineral oil mixture (8:2 by volume). Compound No. 53 (1- 468) at of 5.0, 50.0 and 500.0 nM, respectively, was added to the cells and the resulting mixture was incubated for either 1 minute or 1 hour.

Then, Sul of radiolabelled adenosine (2.5 uci initial concentration of 1 uM) were added to the cell suspension and the mixture was incubated for 10 seconds. concentrations Cells were then centrifuged for 15 seconds at 13,000 rpm and the cell pellets were measured for radioactivity.

Figure 4 depicts inhibition of adenosine transport 53 (1-468) and Figure 5 depicts inhibition of adenosine transport with 1 hour preincubation with compound No. 53 (1-468). with 1 minute preincubation with compound No.

Example 6 Effect of an AICA Riboside Analoq (Series ,1 on Adenosine Release from Isolated Cells A Series II (C-2)-substituted AICA riboside analog was compared with AICA riboside itself for its ability to influence adenosine release from coronary endothelial cells. In this experimental model the cells were treated with 50 pH of the test compound and incubated for 16 hours at 37°C. saline Cells were then washed with phosphate-buffered and medium (to inhibit glycolysis), 50 pm antimycin A (to inhibit oxidative phosphorylation) and 20 pM deoxycoformycin (to inhibit adenosine utilization by adenosine deaminase). This treatment was designed to simulate ischemic condition by inducing net ATP breakdown. resuspended in standard culture containing no glucose Media was then processed for HPLC. Adenosine values are given in Table VI.

TABLE VI Extracellular Adenosine Compound No. Levels M Increase (%) Control 1.42 i 0.17 ---- (1-110) 1.64 i 0.12 15.5 (1-240) 2.79 i 0.19 96.5 xam e 7 Effect of AICA Riboside Analogs (Series II) on adenosine Kinase Activity Inhibition of enzyme activity was determined using a 0.1 ml assay mixture containing 50 mM Tris-maleate, pH 7.0, 0.1% (w/v) BSA, 1 mM ATP, 1 mM MgCl2, 0.5 nu [U-1‘c] adenosine (500 mCi/mmol) and 0.1 pg of purified pig heart Different of test incubated in the assay mixture for 20 37°C. 20 #1 portions were removed and spotted on 2 cm pieces of adenosine kinase. concentrations compound were minutes at From each reaction mixture, whatman DE81 filter paper. The papers were then washed to remove [1‘C] adenosine in 1 mM ammonium formate followed by deionized water and finally 95% ethanol. The papers were dried, and [14C] AMP measured by scintillation counting.

Activities were determined from the amount of [1‘C] AMP formed.

The results are shown in Table VII.

EABLEVVII Compound No. ;g5°1gfll (1-110) > 5000 (1-395) 8 (1-535) 23 (1-551) 40 Example 8 Effect of AICA Riboside Analogs (Series V) on Adenosine Utilization in Isolated Cells Series IV 2'—substituted AICA riboside analogs were tested for their ability to inhibit adenosine utilization in human B lymphoblasts. In this assay, cells were preincubated with the test compound at a concentration of pm, so pm or soo pM together with [33]-adenosine (1 pm) Inhibition of adenosine the extracellular concentration of [3H] adenosine measured by scintillation for a period of 10 minutes. utilization was determined from counting following separation of the nucleosides by TLC.

The results from a comparison of 2‘-O—methyl (Compound No. 20) 2'—0—ethyl 34) and 2'-O-n-butyl (Compound No. 32) analogs of AICA riboside compared to AICA riboside Hypoxanthine and inosine levels were also measured.

(Compound No. are shown in Figure 2.

The of these AICA riboside hypoxanthine and inosine levels (also shown in Figure 2) effects analogs on mirror those effects on adenosine levels suggesting an augmented influence on adenosine utilization mediated by inhibition of adenosine deaminase. This interpretation is supported by direct measurement of the ability of the analogs to inhibit the isolated adenosine deaminase.

Inhibition of adenosine deaminase activity was determined spectrophotometrically using a 1 ml assay mixture containing 50 mM potassium phosphate, pH 7.0, 1 mM alphaketoglutarate, 15 units glutamic dehydrogenase, 0.125 mM NADH, 80 pM and 0.002 units of calf intestinal adenosine Different concentrations of the test compounds were incubated in the adenosine musosa deaminase. assay mixture for 10 minutes at 37°C. The reaction was monitored continuously for oxidation of NADH from the change in absorbance at 340 nm.

The results are shown in Table VIII.

TABLE V1;I Compound No. ;g5&_ug1L 1 (1-110) >5ooo o (1—1aa) 1400 (1-250) 510 (1-262) 175 Example g Effect of AICA Riboside Analogs on Inhibition of Platelet Aggregation in Human Whole gloog The ability of preferred AICA riboside-analogs to inhibit platelet aggregation was examined in human whole blood. Whole blood was drawn from healthy donors and collected in 0.1 pvol. of sodium citrate to prevent coagulation. Platelet aggregation was measured by the impedance technique using a Whole Blood Aggregometer. The test compounds were incubated in whole blood for 10 minutes at 37°C and 10 pM adenosine was added 5 minutes before eliciting aggregation. addition of ADP (6-25 uM) Aggregation was induced by at the minimum concentration inducing full aggregation in untreated controls.

The results are shown in Table IX. zgsrn Ix Series Comgougd No. LQSOLQML 1 (1-110) 2700 I 4 (1-122) zoo 23 (1-343) 38 28 (1-348) 180 29 (1-349) 90 51 (1-466) 193 52 (1-457) 430 53 (1-468) 150 56 (1-48?) 75 59 (1-506) 7o 61 (1-509) 171 71 (1-S62) 4o 72 (1-S63) 300 II 27 (1-395) 950 43 (1-432) 520 IV 32 (1-262) 350 Example 10 Enhanced Oral Bioavailabilitv and Half-Life of AICA Riboside Analogs Certain AICA evaluated for enhanced oral bioavailability in fasted adult beagles. preferred riboside analogs were AICA riboside analogs were given as a 10 mg/kg IV bolus via a cephalic leg vein and as a 20 mg/kg solution administered via a stomach tube. Heparinized blood and urine were collected at selected intervals over 24 hours. Each sample was chilled, centrifuged at 4°C and frozen prior to HPLC analysis.

The results are shown in Table X. rec ' ‘cal Mode of Stab e An ina The AICA riboside analog (1-468) was evaluated for its ability to prevent cumulative cardiac dysfunction associated with repeated episodes of demand-induced ischemia. Anesthetized male dogs were instrumented to measure regional myocardial wall thickening during right atrial pacing in the presence of a stenosis of the left anterior descending artery (Young & Mullane Am. J.

Physiol. In Table XIA, the effects on wall thickening and arterial pressure of six repeated in press (1991)). episodes of pacing in animals treated with a continuous IV infusion of 50 pg/kg/min of the test compound administered post—pace #1 are compared with saline-treated control animals. In Table XIB, the change in heart rate and mean arterial pressure in the post-pace rest period are listed, demonstrating wall that preservation of thickening occurred in the absence of significant hemodynamic effects.

TABLE XIA % of NON-ISCHEMIC WALL THICKENING Pace £ saline (N = 9) Compound No. 53 (n = 6) 1 41.6 1 2.6 49.5 1 6.5 .7 1 4.6 46.7 1 7.0 .8 i 5.6 54.2 1 9.4* .5 i 5.5 48.1 i 7.6* .8 1 5.6 47.5 _+_ 8.2* .4 i 6.0 42.1 3 7.04: * P < 0.05 vs. saline Example 1g Effect of AICA Riboside Analogs (Series 1) in an Experimental stroke Model ' ' The ability of Series 1 (N-4) substituted AICA riboside analogs to effect hippocampal pyramidal cell In this test, male Mongolian gerbils were anesthetized with 2-3% halothane exposed. survival in a gerbil stroke model was evaluated. in N20:o2 and the common carotid arteries Ischemia was then induced by bilateral occlusion of both common carotid arteries for 5 minutes. Seven days following the ischemic insult, brains were removed and processed for histology. shows the effect of pretreatment of the gerbils with 500 (1- The data presented in Figure 3 mg/kg of the AICA riboside analogs (Compound Nos. 186) or 11 (1-226)) or with saline, as a control.

Example A Egepagatiog of 5-Amino-(2,3.5-tgiacety;-beta-D- ribofuranosyl)imidazolecarboxamide (Compound No. 2 (1-111)) AICA riboside (50 g) (450 ml) and then cooled in an ice bath. was dissolved in pyridine Acetic anhydride (80 ml) was added and the ice bath removed. The reaction mixture was stirred for 3 hrs. TLC on silica gel, eluting with 9:1 methylene chloridezmethanol, showed the reaction to be complete. Methanol (5 ml) was added to neutralize unreacted acetic anhydride. The solvents were removed by evaporation under high vacuum (bath temperature less than °C). The residue was coevaporated with dimethylformamide (3 x 150 ml). The residue was crystallized from ethanol using seed crystals. The yield of the triacetate 62 g of white solid; melting point 128- 129°C.

NMR (DMSO-d6) 6 ppm 2.05-2.15 9H, -CH3), 4.3 3H, 4'-CH, 5'-CH2), 5.3 1H, 3'-CH) 5.55 (t, 1H, 2'-CH), 5.87 (d, 1H, 1'-CH), 5.9 (broad 5, 2H, 5- NH2), 6.7-6.9 (broad d, 2H, 4-NH2), 7.4 (5, 1H, 2-CH) (ZS. (broad s, (m, The preparation of this compound is also described in U.S. Patent No. 3,450,693 to K. Suzuki & I. Kumoshiro (1969); See also Chem. Abs. 1;:816982 (1969).

EXETHELE E Preparation of N5—dimethvlaminomethvleneamino-beta—D- ribofuranosylimidazole-4—carboxamide (Compound No. 7 (l-164)) Dissolved 2',3',5'-tri-O-acetyl AICA riboside (10 g) in dimethylformamide (30 ml) dimethyl acetal (20 ml). The reaction mixture was allowed to stir overnight. and dimethylformamide TLC on silica gel, eluting with 9:1 methylene chloride:methanol, showed that the reaction was complete by absence of starting material. The solvent was removed by evaporation under high vacuum (bath temperature than 40°C). The cyclohexylamine and stirred overnight. less residue was dissolved in The solvent was removed by evaporation under reduced pressure and the residue was crystallized from ethanol. white solid, melting point 173-175°C.

NMR (MeOH-d4), 6 ppm 3.0-3.05 (25, 6B, N(CH3)2), 3.75 (m, 2H, 5'-CH2), 4.0 (g, 1H, 4'-CH), 4.2 (t, 1H, 3'-CH), 4.35 (t, 1H, 2'-CH), 5.8 (d, 1H, 1'-CH), 7.7 (s, 1H, 2- CH), 8.25 (S, 1H, 5-N=CH-N ) Yield was 4.6 g of xam e C Preparation of 5-Amino—1—beta—D—ribofuranosylimidazo;e- —N-(cvclopentvl)carboxamide (Compound No. 10 (1-186)) The literature procedure of P.C. Srivastava, R.W.

Mancuso, R.J. Rosseau and R.K. Robins, J. Med. Chem. 17(11), 1207 (1977) was followed to synthesize N- succinimidy1amino(2,3,5-tri-O—acetyl-fi-D- ribofuranosyl)imidazolecarboxylate No. 4"). Intermediate No. 4 (3.9 g) methylene chloride (60 ml). Cyclopentylamine (0.8 ml) was added and the solution was stirred overnight. TLC on silica, eluting with 9:1 methylene chloridezmethanol, ("intermediate was dissolved in showed the reaction was complete by absence of starting material. The solvent mixture was extracted ‘with 5% hydrochloric acid solution (100 ml), saturated sodium bicarbonate solution (100 ml) and water (200 ml). The organic layer was dried over sodium sulfate and evaporated under reduced pressure to give 3.1 g of yellow foam. The acetyl groups were removed by dissolving the 3.1 g of foam in methanol (70 ml) and cooling in an ice bath. Ammonium hydroxide (60 ml) was added and the ice bath was removed.

After 2% hours stirring, TLC on silica gel, eluting with 9:1 methylene ch1oride:methanol, showed all material was gone. The solvent was evaporated under starting reduced pressure to give a residue which was purified on a silica column, chloride:methanol. eluting with 9:1 .and 6:1 methylene Fractions which were alike by TLC were pooled and evaporated under reduced pressure to yield 1.1 g of white foam crystallized from methanol-ethyl acetate, melting point 158-160°C.

NMR (DMSO-d6), 6 ppm 1.4-1.9 (m, 8H, -CH2-CH2-), 3.5 (m, 2H, 5'-cuz), 3.9 (d, 1H, NH-CH ), 4.0-4.35 (m, 3H, 2',3',4'-ca), 5.15-5.4 (m, 3H, 2',3',5'-OH), 5.45 (d, 1H, '-ca), 5.9 (broad s, 2H, -NR2), 7.1 (d, 1H, -NH-), 7.3 1H, 2-CH).

Example D Preparation of 5—Amino—1-beta-D-ribofuranosvlimidazole- (1-232)) This compound was prepared following the procedure —N-(cvclopropvl)carboxamide jcompound No. described in Example C except cyclopropylamine (0.5 ml) was substituted for cyclopentylamine (0.8 ml). The yield starting with 6.2 g of intermediate No. 4 (the succinate ester) was 2.3 g.

NMR (DMSO-d6) 5 ppm 0.5 (m, 4H, CH2-CH2) 2.7 (m, 1H, N—CH ), 3.6 (m, 2H, 5'-CH2), 3.8-4.3 (m, 3H, 2',3',4'- CH), 5.15-5.4 3H, 2',3',5'-OH) 5.45 (d, 1H, 1'-CH), .9 (s, 2H, NH2), 7.2 (s, 1H, 2-ca) 7.4 (d, 1H, 4-NH).

Preparation of 5—Aminobeta-D—ribofuranosy;imidazo;e- 4—N-(benzv1)carboxamide (Compound No. 11 (1-226)) (10 g) was suspended in dimethylformamide (100 ml) and dimethylformamidedibenzylacetal (25 ml). The TLC on eluting with 6:1 methylene chloridezmethanol, Inosine resulting mixture was stirred at 70°C overnight. silica, showed completion of reaction. Solvent was removed by The remainder was dissolved in ammonium hydroxide (130 ml). evaporation at reduced pressure.

The mixture was stirred overnight, then evaporated under reduced pressure.

Ethanol (80 ml) was added to the residue and the resulting mixture The collected by Yield of 1—benzylinosine was 10.5 g which was was warmed. solid was filtration. characterized by NMR.

The intermediate, l-benzylinosine (10.5 g), was dissolved in ethanol (1.0 L) and 3 M sodium hydroxide solution (140 ml). This solution was refluxed for 3 hours. TLC on silica showed the reaction was complete.

The solvent was removed by evaporation under reduced pressure. The residue was chromatographed on a silica gel eluting with 6:1 methylene chloridezmethanol.

Fractions were collected which were similar by TLC and column, concentrated until crystals appeared. Yield was 7.4 g of the above-identified compound as a white solid, melting point 178-179°C. ' _ mm (omso-as) 6 ppm 3.6 (m, 2H, 5'-CH2) 3.85-4.35 (m, 3H, 2',3',4'-CH), 4.4 (d, 2H, N-CH2), 5.15-5.4 (m, 3H, 2',3',5'-OH), 5.5 (d, 1H, 1'-CH), 5.9 (broad S, 2H, 5- NH2), 7.2-7.4 (m, 6H, 2-CH, CGHS) 7.95 (t, 1H, NH).

Example E Preparation of 5-AminoB-D-ribofuranosvlimidazole- -carboxvlic acid methvl ester (Compound No. 14 (1-260)) -amino(2,3,5-tri-Q-acetyl-fi-D-ribofuranosyl)— imidazole-4—carboxylic (3.85 g, 10 mmol) dissolved in 40 ml tetrahydrofuran and cooled to 0°C. An acid was excess of diazomethane in ether was added and the mixture Acetic acid was added to destroy excess diazomethane and the mixture was evaporated warmed to room temperature. to dryness. The residue was purified by chromatography The judged by silica thin layer chromatography (TLC) using the above system, were combined and evaporated to yield 1.2 g of a white foam. This was dissolved in 40 ml of methanol containing 20 mg of sodium methoxide and stirred for 30 minutes. Silica TLC, eluting with 6:1 methylene chloride:methanol, showed no remaining on silica gel, eluting with 7:3 ethyl acetatezhexane. major product fractions, starting material and a new slower—moving product spot.

The reaction was neutralized with Dowex 50 (H+) resin and evaporated to yield 0.64 g or the desired product as a white foam. IR (KBr):l725 cm” (-CO-OCH3).

NMR (DMSO-d6): 6 ppm, 3.65 (s, 3H, CH3), 3.8 (m, 3H, '-CH and 5'-CH2), 4.1 (m, 1H, 3'-CH), 4.2 (m, 1H, 2'- CH), 5.5 (d, 1H, 1'-CH), 8.0 (s, 1H, 2-CH).

Example G Ereparation of 5—Amino—5'-su1famovlB-D—ribofuranosvl- imidazolecarboxamide (Compound No. 15 (1-261)) To,a solution of 2',3'-isopropylidene-AICA-riboside (2.98 g, 10 mmol) in dry N,N-dimethylformamide (25 ml), sodium hydride (300 mg, 80% dispersion in oil) was added over a period of 10 min. After the evolution of hydrogen gas had ceased, the flask was immersed in an ice bath and the mixture was stirred for 30 min. A solution of sulfamoyl chloride (1.3 g, 11 mmol) in dry tetrahydrofuran (20 ml) was added slowly. (silica gel, TLC of the reaction mixture solvent 9:1 methylene chloridezmethanol) indicated presence of some starting material. An additional 200 mg of sulfamoyl chloride in tetrahydrofuran (10 ml) was added and the resulting mixture stirred for one hour. Methanol (1 ml) was added and solvent was evaporated under high vacuum. The residue chromatographed over silica gel, eluting with a ‘mixture of methylene (9:1).

Fractions showing identical TLC patterns were Yield was 1.5 chloridezmethanol Several fractions were collected. pooled and evaporated to a glassy product. g.

H-NMR (nnso-as) 5 ppm, 1.25 and 1.55 (25, en, C(CH3)2), 4.1 (d, 2H, 5'-cnz), 4.25-4.35 (m, 1H, 4'-ca), 4.8-4.9 and 5.1-5.2 (2m, 2H, 2'-CH and 3'-CH), 5.8 (d, 1H, 1'-CH), 5.9 (5, 2H, 5-NH2), 6.65-6.95 (br. d, 2H, CONH2), 7.35 (5, 11-1, 2-CH), 7.7 (s, 2H, SOZNI-I) The NMR data conformed to the structure of 5-amino-2',3'- isopropylidene5-ribofuranosyl—5'-sulfamoylimidazole— 4-carboxamide. This intermediate product was used in the following deblocking step without further purification or isolation.

B. Preparation of 5-Amino-5'-sulfamovl—1-B-D- ribofuranosyl-imidazolecarboxamide lcompound No. 15 11-261)) The compound from the preceeding preparation was dissolved in 60% formic acid (20 ml) solution was stirred at room temperature for 48 hours. and the resulting The solvent was removed by evaporation under high vacuum.

The residue was coevaporated with water. 5 . Yield was 1.0 g of the above-identified product, melting point 174-175°C.

H-NMR (DMSO-d6) 6 ppm 3.9-4.3(m, 5H, 2'-ca, 3--ca, 4'-CH and 5'-CH2), 5.4 and 5.5 (Zd, 2H, 2‘-OH and 3'-OH), .5 (d, 1H, 1'-CH), 5.8 (br.s, 2H, 5-NH2), 6.6-6.9 (br.d, 2H, CONH2), 7.3 (s, 1H, 2-CH) and 7.6 (S, 2H, SOZNH2).

The product was crystallized from aqueous ethanol.

Example fl greparation of 5'-Amino-5'-deogy-AICA-riboside lcompound No. 21 (1-227)) A. Preparation of 5'-Azido-5'-deoxv-AICA-riboside were Excess formic acid was removed by evaporation under high vacuum. coevaporated with water (3 x 25 ml) solid product.

The residue was to obtain a semi- This product was crystallized from aqueous .0 g, of the product, melting point 138-139°C.

‘H NMR (DMS0-d6) 6 ppm 3.55 (d, 2H, 5'-CH2), 3.95 (br. s, 2H, 3'-CH and 4'-CH), 4.2-4.4 (m, 1H, 2'-CH), 5.35 and .50 (2d, 2H, 2'-OH and 3'-OH), 5.55 (d, 1H, 1'-CH), 5.75- ethanol. Yield was above-identified .9 (br. s, 2H, 5-NH2), 6.6-6.9 (br. d, 23, connz) and 7.35 (s, 1H, 2-CH). IR (KBr) cm": 3400-aooo (br. NH2, CONH2, on, etc.), 2150 (s, N3) 1640 (CONH2).

. Preparation of 5'—Amino-5'—deoxy-AICA-giboside A solution of 5'-azido-5'-deoxy-AICA-riboside mg) (the product of step (A)) (40 ml) hydrogenated in a Parr apparatus with palladium on carbon (5%) (100 mg) as the hydrogenation catalyst at 40 psi for 60 min. The catalyst was removed by filtration of the reaction mixture through a celite pad. The clear filtrate was evaporated to dryness. The product was crystallized from boiling ethanol. (aoo in methanol was Yield was 650 mg of the above- identified product, melting point 188-189°C.

‘H-NM (D20) 5 ppm, 2.7 (d, 23, 5‘—CH2), 3.3-4.4 (3m, 2'-ca, 3'-ca and 4'-cu), 5.4 (d, 1H, 1'-ca) and 7.3 1H, 2-CH). IR (KBr) cm‘1: 3500-aooo (br. on, NH2, counz, etc.), 1540-1545 (br.s. CONH2).

H, (5.

Example 1 Preparation of 5—Amino{2-O-methvl-B-D-ribofuranogyll; (Compound No. /30 (1-188)) and -Amino(3-O-methyl-B-D-ribofuranosvl)imidazole 22 (1-243)) -Amino-1—B—D—ribofuranosylimidazolecarboxamide (5.2 g, 20 mmol) was dissolved in 40 ml hot dimethylform— amide and diluted with 70 ml methanol containing 35 mg tin(II) A solution of 0.1 mol of diazomethane in 200 ml of ether was added in portions over mg of tin(II) chloride The resulting mixture was filtered imidazolecarboxamide carboxamide (Compound No. chloride dihydrate. 45 min. After each addition, dihydrate was added. and evaporated to give a syrup. The syrup was dissolved in 25 ml of methanol and upon cooling yielded crystalline -amino—1-(2-O-methy1-B-D-ribofuranosyl)imidazole carboxamide which was collected by filtration and dried.

Yield was 1.2 g, melting point 114-117°C.

‘H NMR (DMSO-dc) (for Compound 20): 6 ppm, 3.3 (s, 3H, CH3), 3.6 (m, 2H, 5'-CH2), 3.9 (m, 1H, 4'-CH), 4.1 (m, 1H, 2'-CH), 4.2 (m, 1H, 3'-CH), 5.2 (d, 1H, 3'-OH), 5.3 (t, 1H, 5'-OH), 5.6 (d, 1H, 1'-CH), 6.0 (br. s, 2H, 5- NH2), 6.7 (br. d, 2H, 4-CONH2), 7.3 (S, 1H, 2-CH).

The supernatant from the above crystallization was concentrated and applied to a 200 ml column of silica gel.

The eluted with 10:1 chloride:methanol (1 L), 8:1 methylene chloride:methanol (500 ml) (500 ml).

The 5:1 and evaporated and residue dissolved in 10 ml of methanol.

Upon cooling this yielded crystals which were collected and dried. column was methylene and 5:1 methylene chloride:methanol eluate contained a major product was Yield was 1.4 grams. By NMR decoupling and exchange experiments the product was shown to be 5-amino- 1-(3-O—methylD-ribofuranosyl)imidazolecarboxamide.

H NMR (DMSO-d6) (for Compound 18): 6 ppm: 3.3 (s, ‘3H, CH3), 3.6 (m, 2H, 5'-cnz), 3.7 (m, 1H, 4'-CH), 4.0 (m, 1H, 3'-CH), 4.4 (m, 1H, 2'-CH), 5.3 (t, 1H, 5'-OH), 5.4 (2d, 2H, 2‘-CH and 1'-CH), 5.9 (br. s, 2H, 5-NR2), 6.7 (br. d, 2H, CO-NH2), 7.7 (S, 1H, 2-CH). fl—[(4-nitrophenyl)methylJcarboxamide (Compound No. 23 (l-343))’ N-Succinimidylamino(2,3,5-tri-Q-acetyl-fi-D- ribofuranosyl-imidazolecarboxylate3 (0.50 g), 4- nitrobenzylamine hydrochloride (210 mg) and triethylamine (0.16 ml) (30 ml) temperature overnight. was washed with then resulting yellow were stirred in chloroform at room The solution saturated sodium bicarbonate solution The tar was chromatographed on silica gel, and water, evaporated under reduced pressure. eluting with 9:1 methylene chloridezmethanol. The collected fractions were monitored by TLC. The like fractions were combined and concentrated under reduced pressure to afford a yellow foam (0.38 g). The foam was dissolved in methanol (20 ml) and methanolic sodium methoxide solution was added (0.3 ml of 0.25 n solution). argon atmosphere for 15 min.

The solution was stirred under an TLC indicated the reaction was complete. The solution was neutralized to pH 6 with ion exchange resin. The resin was filtered and the solution concentrated under high vacuum to yield a yellow foam (0.23 g).

‘H NMR (DMSO—d6) 5 ppm, 3.6 (m, 2H, 5'-CH2) 3.9-4.3 (m, 3H, 2'-CH, 3'-CH, 4'-CH), 4.5 (d, 2H, -CH2-CGH4-N02), .2-5.4 (br., 3H, 2'-on, 3--on, 5'-on), 5.5 (d, 13, 1'- “‘Srivastava, P.C., J. Med. Chem. fl:1207 (1974).

CH), 6.0 (br. 5, 2H, 5-NH2), 7.3 (S, 1H, 2-CH), 7.4-8.2 (ABQ, 4H, -CGH4-N02), 8.3 (t, 1H, 4*CONH).

‘Example K Ereparation of 5-AminoB-D-ribofuranosvlimidazoleN- 112-chlorophenyljmethvllcarboxamide jcompound No. 24 (1-354)) This compound was prepared according to the procedures described in Example J for the 4-p-nitrobenzyl derivative, substituting 2—chlorobenzylamine for #- nitrobenzylamine hydrochloride.

H NMR (DMSO-d6) 6 ppm, 3.6 (m, 2H, 5'—CH2), 3.9-4.3 (m, 3H, 2'- CH, 3'-CH, 4‘-CH), 4.4 (d, 2H, -CH2-O-Cl), 5.1- .4 (br., 3H, 2'-on, 3‘-OH, 5'-OH), 5.5 (d, 1H, 1'-CH), 6.0 (br.s., 2H, 5—NH2), 7.2-7.4 (m, 4H, —C6H4-Cl), 8.0 (t, 1H, 4-CONH).

Example L Preparation of 5-aminofl-D-ribofuranosvlimidazo1e—4—N- J12,4-dich1oroohenv1)methvl1carboxamide jcompound No. 25 (1-360)) This compound was prepared according to the procedures described in Example J for the 4-p-nitrobenzyl derivative, substituting 2,4—dichlorobenzylamine for 4- nitrobenzylamine hydrochloride.

‘H NMR (DMSO—d6), 5 ppm, 3.6 (m, 2H, 5'-CH2), 3.9- 4.3 (m, 3H, 2‘-CH, 3‘-CH, 4'-CH), 4.4 (d, 2H, —CH2-C6H3- C12), 5.2-5.4 (m, 3H, 2'-on, 3'-on, 5‘-OH), 5.5 (d, 1H, 1'- ca), 6.0 (br. s, 2H, 5-NR2), 7.2-7.6 (m, 3H, -CGH3-C12), 8.1 (t, 1H, 4-CONH—).

Example g Preparation of 5-aminothioB-D-ribofuranogyl imidazolecarboxamide (Compound No. 27 (10)) To 10 ml of 80% formic acid was added 400 mg of 5- amino-2—thio(2,3isopropylidene-fl-D-ribofuranosyl)- imidazole-4—carboxamide.‘ The resulting mixture was stirred for‘ 1 hour‘ at room ‘temperature. Silica TLC, eluting with 4:1 methylene chloride:methanol, conversion of staring material to one major product. mixture was evaporated to dryness, showed The dissolved in 5 ml of methanol and applied to a 50 ml column of silica gel. The column was eluted with methylene ch1oride:methanol (5:1).

The major product, as determined by TLC, was collected and evaporated to dryness. The residue was dissolved in 3 ml of hot methanol and crystallized upon cooling. Yield was 150 mg of the above-identified product, melting point 205- 208°C. in mm (DMSO-d6), 6 ppm 3.6 (m, 2H, 5'-caz), 3.8 (m, 1H, 4'-CH), 4.1 (m, 1H, 3'-CH), 4.5 (m, 1H, 2'-CH), 5.1 (d, 1H, 2' or 3'-OH), 5.2 (d, 1H, 2' or 3'-OH), 5.7 (t, 1H, 5'-OH), 6.3 (d, 1H, 1'-CH), 6.4 (br. s, 2H, 5-NR2), 6.9 (br. s, 211, 4-CONH2), 11.1 (br. s, 1H, 5'-sa). carbon tetrachloride stirred in (38 ml) The solution was diluted with methanol (15 ml), then concentrated under reduced pressure. and were dimethyl formamide hours.

The resulting yellow tar was chromatographed on silica gel, eluting with ‘Preparation described in T. Miyoshi, S. Suzaki, A. Yamazaki. Chem.

Pharm. EL, gs (9):2089-2093 (1976). :1 methylene chloride:methano1. The like fractions were combined and concentrated under reduced pressure to afford a purple foam. The presence of triphenylphosphine oxide, as determined by 1H NMR, necessitated a second chromato- graphic step as above. Yield was 0.43 g of a white foam.

H NMR (nnso-d6), 6 ppm 3.7-3.9 (m, 2H, 5'-CH2), 4.0- 4.4 (m, 3H, 2'-CH. 3'-CH, 4'-CH), 5.4-5.5 (m, 2H, 2'-on, 3'-on), 5.6 (d, 1H, 1'-ca), 5.9 (br. s, 2H, 5-NH2), 6.7- .9 (hr. d, 2H, 4-CONH2), 7.3 (S, 1H, 2-CH).

Exam e O -imidazole carboxamide (Compound No. 34 carboxamide (Compound No. 31 (1-251)) A solution of approximately 30 mmol diazoethane in 40 ml of ether was prepared by slow addition of 7 g (44 mmol) of 1-ethylnitro—1-nitrosoguanidine to a mixture of 8 g 9 ml water and 60 ml of ether followed by distillation. 3.2 g (12 ribofuranosylimidazo1e—4-carboxamide (AICA riboside) in 35 of potassium hydroxide, This was slowly added to a solution of mmol) of 5-aminofi-D- ml dimethylformamide containing 50 mg of tin(II) chloride dihydrate. methanol was added to maintain solubility.

During the addition approximately 20 ml of The reaction was filtered to remove a trace precipitate and evaporated to a yellow syrup. Thin layer chromatography on silica gel using methylene chloride/methanol (3:1) showed a major product spot moving faster than AICA riboside. The syrup was chromatographed on silica gel using methylene chloride/methanol (8:1) collecting the major product as determined by TLC. The evaporated to a white foam. appropriate fractions were This was dissolved in 7 ml of methanol. Upon cooling to 4°C the mixture crystallized to yield 160 mg of 5-amino(2-O-ethyl—fi-D-ribofuranosyl) imidazo1e—4-carboxamide (Compound No. 34 (1~250)) confirmed by NMR decoupling and exchange experiments.

‘H NMR rnnso-as) (for Compound No. 34) 5 ppm, 1.05 (t, 3H, cal), 3.3-3.6 (m, 4H, 2'-OCH:-, 5'-cnz), 3.9 (m, 13, 4'-CH), 4.1-4.3 (m, 2H. 2'~CH, 3'-CH), 5.15 (d, 1H, 3-03), .25 (t,.1H, 5'-OH), 5.55 (d, 1H, 1'-CH), 6.0 (br.s, 2H, -NH2), 6.6-6.9 (br.d, 2H, 4-CONH2), 7.3 (3; 1H, 7-CH).

The supernatant from the above crystallization was cooled overnight at -12°C yielding a second crop of crystals, 0.58 g, which by NMR decoupling and exchange experiments was shown to be mostly 5—amino—1-(3—0-ethyl- fi—D—ribofuranosyl) imidazole-Q-carboxamide (compound No. 31 (1-251)).

‘H NM (DMSO~da) (for Compound No. 31): 6 ppm, 1.1 (t, 3H, CH3), 3.4-3.7 (m, 4H 3'-OCH:-, 5'-CH2), 3.85 (m, 1H, 4'-CH), 4.0 (m, 1H, 3‘-CH) 4.4 (q, 1H, 2-CH), 5.25 (t, 1H, 5'-OH), 5.35 (d, 1H, 2'-OH), 5.45 (d, 1H, 1'-CH), 5.9 (br.s, 2H, 5-NH2), 6.6-6.9 (br.d, 2H, 4-CONH2), 7.3 (5, 1H, 1-CH): The major impurity was identified as the 2'-O- ethyl isomer.

Examgle P gggparation of 5—aminoI2n-butvl-Hribofuranosvl) imidazole—4-cgrboxamide and 5—amino—1-(3-O—n-butv1-B-D- ribofuranosvlj imidazolecarboxamide (Compound Nos. 32 1- 62 _a d 33 -263 -Amino—1D-ribofuranosylimidazolecarboxamide (2.50 g, 10.0 mmol) and tin(II) chloride hydrate (35 mg) were dissolved in dimethylformamide (40 ml) and methanol (30 ml). A solution of 0.1 ml of diazobutanes in 150 ml of ether was added in portions. Halfway through the Diazobutane was prepared by treatment of 16.59 of N-n‘rtroso-N-n- butylmethane [Wi|ds, AL. and Meeder, AL, SOC 1;; (1948)] in ethyl ether (mom!) with potassium hydroxide (55 g) in water (60 ml). The ethereal diazobutane was used without distillation. addition, more tin mg). ing material stayed in solution.

(II) chloride hydrate was added (35 Methanol was added, as needed, to ensure the start- The mixture was stirred "for 1 hr, then concentrated under reduced pressure to give Analysis of the oil by 1H NMR showed mostly N- butylethylcarbamate. an oil.

The oil was stirred with hexane and decanted to remove the N-butylethylcarbamate. The result- ing tar was chromatographed on silica gel using 6:1 The appropriate fractions were combined and concentrated under ‘H NMR analysis showed a mixture of 2' and 3' butyl ethers. methylene chloride:methanol as eluting solvent. reduced pressure to give a pink foam.

HPLC analysis showed a 56:28 mixture. The solid was dissolved in isopro— panol (2 ml) and cooled. and dried to give 63 mg.

The resulting solid was filtered HPLC analysis showed a 77/18 mixture. 1H NMR decoupling and exchange experiments showed the major product to be the 2'-O-n-butyl ether.

‘H NMR (DMSO-d6) (for Compound No. 32): 5 ppm, 0.8- 1.5 (m, 7H, -CH2CH2CH3), 3.3-4.2 (m, 7H, 2'-OCH2-, 2'-CH, 3'-CH, 4'-CH, 5'-CH2), 5.1 (d, 1H, 3'-on), 5.3 (t, 1H, 5'- OH), 5.6 (d, 1H, l'-CH), 6.0 (br.s, 2H, 5-NR2), 7.6-7.8 (br.d, ZN, 4-CONH2), 7.3 (S, 1H, 2-CH).

The supernatant from the above crystallization was concentrated under reduced pressure to give 125 mg of a 1H NMR showed the major pink foam. HPL analysis showed a 14/71 mixture. decoupling and exchange experiments product to be the 3'-O-n-butyl ether. —1H NMR (DMSO-d6) (for Compound No. 33): 5 ppm, 0.8- 1.5 (m, 7H,-CH2CH2CH3), 3.4-4.4 (m, 7H, 3'-ocaz-, 2'-CH, 3'-ca, 4'—CH, 5'-CH2), 5.2 (t, 1H, 5'-on), 5.3 (d, 1H, 2'- on), 5.4 (d, 1H, 1'-ca), 5.9 (br.s, 2H, 5-NH2), 6.6-6.8 (br.d., 2H, 4-CONH2), 7.3 (s, 1N, 7-CH).

Example Q _ Preparation of 5—amino—1—B-D-ribofuranosvlimidazcleN- [(3-nitrophenyl)methyl]carboxamide (Compound No. 28 11-348)) This compound was prepared according to the procedures described in example J for the 4-p-nitrobenzyl derivative, substituting 3-nitrobenzylamine hydrochloride for 4-nitrobenzylamine hydrochloride.

H NMR (DMSO-d6) 6 ppm, 3.5 (m, 2H, 5'-CH2), 3.9-4.3 (m, 3H, 2'-CH, 3'-CH, 4'-CH), 4.4 (d, 2H, -CH2- N02), 5.2- .4 (br., 3H, 2'-OH, 3'-OH, 5'-O), 5.5 (d, 1H, l‘-CH), 6.0 (br.s., 2H, 5-NH2), 7.4 (s, 1H, 7-CH), 7.6-8.2 (m, 4H, -C6H4Cl), 8.3 (t, 1H, 4-conn).

Example R Preparation of 5—aminoB-D-ribofuranosvlimidazoleN- II4—Ch1orophenvl)methvllcarboxamide (Compound No. 22 11-349)) This compound was prepared according to the procedures described in Example J for the 4-P—nitrobenzyl derivative, substituting 4-chlorobenzene amide for 4- nitrobenzylamine hydrochloride.

H NMR (DMSO—d6) 6 ppm, 3.6 (m, 2H, 5'-CH2), 3.9-4.3 (m, 3H, 2'-CH, 3'-CH, 4'-CH), 4.4 (d, 2H, -CH2-CGH4-Cl), .2-5.4 (br., 3H, 2'-OH, 3'-OH, 5'-OH), 515 (d, 1H, 1'-CH) .9 (br.s., 2H, 5-NH2), 7.3-7.4 (m, SN, -C6H‘Cl), 7-CH), .1 (t, 1H, 4-CONH).

Example S Preparation of 5-aminoE-D-ribofuranosvlimidazoleN- J14-methvlphenvl)methvllcarboxamide (Compound No. 30 11-388)) This the procedures described in Example J for the 4-p-nitrobenzyl compound was prepared according to derivative, substituting 4-methylbenzylamine for 4- nitrobenzylamine hydrochloride.

‘H NMR (DMSO—d6) 5 ppm, 2.2 (s, 33, -Cefl‘-CH3), 3.6 (m, 2H, 5'-cnz), 3.9-4.3 (m, 5H, 2'-CH, 3--cu, 4'-ca, - CH2- -CGH4-CH3), 5.2-5.4 (br., 3H, 2'-OH, 3'-OH, 5'-0H),» .5 (d, 1H, 1'-CH), 5.9 (br.s., 2H, 5-NH2, 7.1-7.2 (M, 4H, -CGH4-CH3), 7.3 (S, 1H, 7-CH), 7.9 (t, 1H, 4-CONH).

Example Preparation of 5-aminoB-D-ribofuranosvl-imida2o1e NI(3-chloronhenvllmethvllcarboxamide (Compound No. 35 (1-355)) This compound was prepared according to the procedures described in Example J for the 4-P-nitrobenzyl derivative, substituting 3~chlorobenzylamine for 4- nitrobenzylamine hydrochloride.

H NMR (DMSO-d6) 6 ppm, 3.5 (m, 2H, 5'-cuz), 3.9-4.3 (m, 3H, 2'-CH, 3'-CH, 4'-CH), 4.3 (d, 2H, -CH2-CGH4-Cl), .1-5.4 (br., 3H, 2'-OH, 3'-OH, 5'-OH), 5.5 (d, 1H, 1'- CH), 6.0 (br.s., 2H, 5-NR2), 7.2-7.4 (m, 4H, -CSH4-Cl), 7.4(s, 1H, 7-CH), 8.1 (t, 1H, 4-CONH).

Example U Preparation of 5-amino(1-piperidinocarbamovl)B-D- 36 I1-207)) This compound in Example J for the 4-p—nitrobenzy1 ribofuranosvlimidazole (Compound No. derivative, substituting piperidine for 4-nitrobenzylamine hydrochloride. The product was crystallized from ethanol to give the above-identified product, m.p. 190-192°C.

H NMR (DMSO-d6) 6 ppm, 1.4-1.7 (M, GB, 3, 4, 5-cnz 3.55 (m, 2H, 5'-CH2), 3.8- 3.95 (m, SH, 2- and 6-CH2 groups of piperidine ring, and 4'-CH), 4.0-4.1 (m, 1H, 3'-CH), 4.25-4.35 (m, 7H, 2-CH) .15 (d, 1H, 2' or 3'-OH), 5.2 (t, 1H, 5'-OH). groups of piperidine ring), E-[Q-methoxybenzyl)carboxamide Com ound No. 39 -3 0 A mixture of the activated succinate ester (0.5 g) (prepared according to Example J), 4-methoxybenzylamine (0‘15 ml) (20 ml) was sirred overnight. TLC indicated completion of the reaction. The solvent was evaporated and the residue was chromatographed and methylene chloride gel column using a mixture of (9:1). The . containing the product were pooled and evaporated. over a short silica fractions The residue thus obtained was dissolved in methanol (20 ml) and the pH was adjusted to about 10 by adding a sodium methylene chloridezmethanol methoxide solution. After stirring the reaction mixture for 45 minutes at room temperature,— the solution was The resin was filtered off, washed with methanol (2 x 2 ml). neutralized with Dowex 50 H+-resin (pH about 6.0).

The combined filtrate and the washings was evaporated and the residue was crystallized from ethanol. Yield was 100 mg, with a mp of 187-188°C.

H NMR (DMSO-d6): 6 ppm, 3.55 (m, 2H, 5'-cnz), 37 (s, 3H, -OCH3), 3.7-4.1 (m, 3H, 2'-CH, 3'-CH, and 4'-CH), 4.35- 4.2 (dd, 2H, -CH2-N-), 5.1-5.4 (3,m, 3H, 2'-OH, 3'-OH, and '-OH), 5.45(d, 1H, 1-CH), 5.9 (hr. 2H, NH2), 6.8-7.2 (m, 4H, aromatic—phenyl), 7.3(s, 17H, C2-H), and 7.85 (t, 1H, C-NH).

Example W Preparation of 5—AminoB-D-ribofuranosylimidazole-4— N(4-dimethylaminobenzyl)-carboxamide hydrochloride jcompound No. 41 Il3)) of hydrochloride (245 mg, 2 mmol) in methylene chloride (25 ml), (222 mg, 2 mmol) was added and the resulting mixture stirred 45 minutes to it was added the To a suspension 4-dimethylaminobenzylamine triethylamine activated succinate ester prepared according to example J (500 mg); temperature overnight. the resulting mixture was stirred at room TLC indicated completion of the reaction. The reaction mixture was evaporated and the residue was chromatographed through a short silica gel column using a mixture of methylene chloride-methanol (9:1). and evaporated to dryness.

Fractions showing the major product were pooled The residue was dissolved in methanol (15 ml) and the pH was adjusted to about 10 using a sodium methoxide solution. After stirring at room temperature for 45 mintues, the solution was neutralized The resin was filtered off and washed with methanol (2 x 5 ml). and the washings were evaporated to dryness. with Dowex 50-resin.

The combined filtrate The residue which was in the form of a foam was dissolved in absolute ethanol (10 ml). about 5 with an ethanolic—HCl solution.

The pH of the solution was adjusted to Solvent was evaporated to dryness and the residue was treated with anhydrous ether. The amorphous solid that separated was collected by filtration and washed with ether (2 x 10 ml), and dried under high vacuum to yield 250 mg. The compound obtained was highly hygroscopic; no melting point could be obtained.

H mm (020) 5 ppm, 3.05 (s, an, N(CH3)2), 3.5 (m, 2H, '-CH2), 3.8-4.3 (3m, 3H, 2'-CH, 3'-CH, and 4'-CH), 4.4 (S, 2H, CH2fN-), 5.5 (d, 1H, 1'-CH), 7.3-7.4 (m, 4H, phenyl), and 7.9 (s, 1H, 2-CH).

Example 3 Preparation of (R)-S-AminoE-D-ribofurano-svlimidazole- 4-N—r2-hvdroxv13.4—dihvdroxvDhenvll ethvllcarboxamide (Compound 42 11-43111 This compound was prepared according to the procedure described in Example J substituting (R)-norepinephrine for 4—nitrobenzylamine hydrochloride and dimethylformamide in place of chloroform as the reaction solvent.

‘Cg-OH), 5.2-5.2 (m, ‘H NMR (nnso-as): 6 ppm, 3.1 - 3.3 (m, 3.5-3.6 (m, 2H, 5'-CH2), 3.8-3.9 (m, 1H, 4'-CH) 1H, 3'-CH) 4.2-4.3 (m, 1H, 2'-CH), 4.4-4.5 (m, 1H, 2' or 3'-OH), 5.2-5.3 (t, 1H, 5'- OH) 5.3-5.4 (m, 1H, 2' or 3'-OH), 5.4-5.5 (d, 1H, 1'-CH), .9 (br. s, 2H, 5-NH2), 6.5-6.8 (m, 3H, aryl of catechol), 7.1 (t, 1H, 4-CONH), 7.3 (S, 1H, 2-CH), 7.2-7.8 (br. 2H, catechol-OH).

H,-CH2-N), 4.0-4.1 (m, 1H, phenyl- and triethylamine (0.61 g) were refluxed in a The reaction mixture was concentrated and The methylene chloride mixture was washed with water and the residue mixed with 40 ml of methylene chloride. saturated sodium bicarbonate and dried over magnesium sulfate. The methylene chloride was evaporated and the residue purified by chromatography on 200 ml of silica gel using a mixture of methylene chloride and methanol (95:5), 0.5 g of isopropylideneD—ribofuranosyl)imidazolecarboxamide.

Treatment of that compound with 80% formic acid for 3 hours at room termperature to remove the isopropylidene yielding 5-aminothiopheny1-l-(2,3—0- group followed by evaporation and purification by silica (9:1) yielded 250 mg of the title compound as a white foam.

‘H NMR (DMSO-d6) 6 ppm, 3.3-3.5 (m, 2H, 5'-CH2), 3.8- chromatography using methylene ch1oride:methanol .9 (m, 1H, 4'-CH)4.0-4.1(m, 1H, 3'-CH), 4.5 (q, 1H, 2'- cn) 5.1 (d, 1H, 2'— or 3' -on), 5.3 (d, 1H, 2'-or 3- - on), 5.7 (t, 1H, 5'-on), 5.9 (d, 1H, 1'-CH) 7.5 (br. s, ‘ Miyosi T., Chem. Pharm. Bull. 2422089 (1976).

E, 4-NR2), 6.7 and 7.1 (br s, 2H, CONH2) 7.1-7.5 (m, 5H, phenyl).

Example Z A.mixture of (1) endoaminonorbornane hydrochloride (240 mg), triethylamine (:'O mg) and methylene chloride was stirred at room temperature for 45 minutes under argon. To it was added activated succinate ester (See TLC indicated Solvent was evaporated and Example J) (750 mg) and stirred overnight. completion of the reaction. the residue chromatographed over silica gel column using a mixture of methylene chloride.methano1 (9:1). Fractions 4 The residue was dissolved in methanol (25 ml) and the pH was adjusted to about 10 with a sodium methoxide solution.

After stirring for 45 minutes at room temperature the containing the product were pooled and evaporated. solution was neutralized with H+ resin (pH approximately 6). The resin was filtered off and washed with methanol.

The combined washings and the filtrate was evaporated and the residue kept under high vacuum to obtain a solid glossy product. Yield was 280 mg.

‘H NMR (DMSO-d6) 5 ppm, 1.1-2.4 (m, 10H, norbonyl), 3.6 (br.M, 2H, 5'-CH2), 3.9 (m, 1H, -N-CH), 4-4.4 (2 m, 3H, 2‘-CH, 3}-CH and 4'-ca), 5.05, and 5.35 (2-d, 2H, 2--on and 3'-on), 5.25 (t, 1H, 5'-on), 5.5 (d, 1H, 1'—CH), 5.9 (hr. 2H, NH2) 6.8 (d, 1H,-NH-CO), 7.25 (S, 1H, 2-CH).

Example 55 Preparation of 5—AminoB-D-ribofuranosvl- imidagole-4—N-[(3—iodopheny1)methy;Jcarboxamide jcompound No. 44 (1-434)) This compound prepared according to the procedures described in Example J for the 4-p-nitrobenzyl derivative, substituting 3-iodobenzylamine hydrochloride for 4-nitrobenzylamine hydrochloride.

‘H NMR (onso-as) 5 ppm, 3.6 (m, 2H, 5'-CH2), 3.9-4.3 (m, 3H, 2'-CH, 3'-CH, 4'-CH), 4.3 (d, 2H,-Cfiz-CGH‘-I), 5.2- .4 (m, 3H, 2'-OH, 3'-OH, 5'-OH), 5.5 (d, 1H, 1'-CH), 5.9 (br.s., 2H, 5-NR2), 7.1-7.7 (m, 4H, -CGH‘), 7.3 (s, 1H, 2- ribofuranosyl)imidazoleN—[(4-nitrophenyl)methyl]- carboxamide (Compound No. 46(1-445)) The compound used in this procedure, 5-amino(5- iododeoxy-2,3-isopropylidene-E-D-ribofuranosyl) imidazole-4—N—[(4-nitrophenyl)methylcarboxamide, was prepared by the same reaction sequence (stopping at step B) described in Example AH for compound 53 (1-468), substituting the 4-N-p-nitrobenzylamide (compound 23 (1- 343)) (1- 349)).

—Amino—1-(5-iodo—5-deoxy-2,3isopropylidene-fi-D- for’ the 4-N—p-chlorobenzylamide (compound 29 ribofuranosyl)imidazoleN-[(4-nitrophenyl) methylcarboxamide (200 mg) was dissolved in 10 ml of 80% formic acid. The solution was stirred at 45°C for 2 The pressure and the resulting residue co-evaporated twice hours. solvents were evaporated under reduced The residue was 6/1 The appropriate with water and twice with methanol. chromatographed on silica gel, using methylene chloride/methanol as eluting solvent. fractions were combined and concentrated under reduced pressure to yield 60 mg of the above-identified compound as a yellow foam.

H NMR (DMSO-d6) 6 ppm, 3.3-3.6 (m, 2H, 5'-CH2), 3.8- 4.4 (m, 3H, 2'-CH, 3'-CH4‘-CH), 4.5 (d, 2H, Cflz-C6H‘NO2), .4-5.5 (m, 2H, 2'—OH, 3'-on), 5.5 (d, 2H, 1'-CH), 5.9 (bros¢' 2H’ 704 (S, C634-N02, 8.3(4,lH,4-CONH-).

H, 2-CH), 7.5-8.2 (m, 4H, e AC Preparation Exam of 5—AminoB-D-ribofuranosvlimidazole carboxvlic Acid. D-Nitrobenzvlthio Ester (Compound No. 47 11-450)) —Amino-1(2,3,5-triacety1D-ribofuranosyl) imidazolecarboxylic acid1 (1.0 g) was dissolved in 8 ml of thionyl chloride under argon with stirring for 10 minutes. The mixture was evaporated under vacuum and the residue was dissolved in 15 ml of tetrahydrofuran contain- ing 2.0 g of p-nitrobenzyl mercaptan. Triethylamine (1.5 ml) was added and the mixture stirred under argon for 20 minutes. The reaction is evaporated to a gum and the residue mixed with 50 ml of methylene chloride and washed with 2 x 25 ml of water. dried over magnesium sulfate and evaporated to a syrup The methylene chloride phase was which was purified by chromatography on silica gel using a mixture of ethyl acetate and methylene chloride (1:1y yielding 500 mg of 5-amino(2,3,5-tri-O-acetyl—B-D- ribofuranosyl)imidazolecarboxylic p- Treatment with sodium methoxide in acid, nitrobenzylthio ester. ml of dry methanol such that a slightly basic pH was maintained until deacetylation was complete (as determined by thin layer chromatography), followed by neutralization with Dowex 50 (H+) and evaporation yielded the desired compound contaminated with a product presumed to be the methyl ester. Purification by chromatography on silica using a mixture of methylene chloride and methanol (9:1) gave 38 mg of the desired compound as a yellow foam.

H NM (DMSO-d6) 6 ppm, 3.5-3.7 (m, 2H, 5'-CH2), 3.9- 4.0 (m, 1H, 4'-CH), 4.2-4.4 (m, 2H, 2'-and 3'-CH), 5.2 (d, 1H, 2'-or 3‘-OH), 5.3-5.5 (m, 2H, ‘ Srivastava, P.C., J. Med. Chem. fl:1207 (1974). ' and 2'-or 3'—oH), 5.6 (d, 1H, 1‘-CH), 6.9 (br. s, 2H, CH), 7.6 and 8.2 (d, 2H, phenyl).

-NH2), 7.4 (5, 1h, 2- Example AD re a at'on of 5-Amino- - -D- ibofuranos N-indolinvlcarboxamide (Compound No. 48 (1-452)l This prepared according to procedures described in Example J for the 4-p-nitrobenzyl derivative, substituting indoline for 4-nitrobenzylamine hydrochloride.

‘H NMR (nmso-:16) 5 ppm, 3.1 (t, 211, indolinyl-CH2), 3.6 (m, 2H, 5'-CH2-), 5.2-5.4 3H, 2'-OH, 3'-OH, 5'- OH), 5.5 (d, 1H, 1'-CH), 6.4 (br.s., 2H, 5-NH2), 6.9-8.1 (m, 4H, indolinyl aromatics), 7.4 (S, 1H, 2-CH). compound was Example AE Preparation of (R)AminoQ-D—ribofuranosylimidazo;e- -N-[;nitrophenyl)ethyl]carboaxamide Lcompound No. 49(1-453)) This compound the procedures described in Example J for the 4-p-nitrobenzyl derivative, was prepared accoridng to substituting (R)nitro-a-methylbenzylamine hydrochloride for 4-nitrobenzylamine hydrochloride.

H NMR (DMSO-d6) 6 ppm, 1.5 (d, 3H, a-methyl on N4- benzyl carboxamide), 3.6 (m, 2H, 5'-CH2), 3.9-4.3 (m, 3H, 2'-CH,,3'-CH, 4'-CH), 5.1 (m, 1H, benzylcarboxamide), 5.1-5.4(m, 3H, 2'-OH 3'-OH, 5'-OH), .5 (d, 1H, 1'-CH), 7.3 (S, 1H, 2-CH), 7.6-8.2 (m, 4H, CGH4-N02), 8.0 (d, 1H, 4-CONH-). methine proton on N4- Bxample AF Preparation of (S)AminoB-D-ribofuranosylimidazo1e— 4-N-r1-(4-nitrophenvl)ethvllcarboxamide Lgompound No. 50(1-459)) This compound the procedures described in Example J for the 4-p-nitrobenzyl was prepared according to derivative, substituting (S)nitro-a-methylbenzylamine hydrochloride for 4-nitrobenzylamine hydrochloride.

‘H NMR (DMS0-d6) 6 ppm, 1.5 (d, 3H, a-methyl on N4- benzyl carboxamide), 3.6 (m, 2H, 5-CH2), 3.9-4.3 (m, 3H, 2'-CH, 3'-CH, 4'-CH), 5.1 (m, 1H, methine proton on N4- benzylcarboximide), 5.1-5.4 (m, 3H, 2'-OH, 3'-OH, 5'-OH), .5 (d, m, 1'-cm 5.9 (br.s., 2H, 5-NH2), 7.4 (s, 1H, 2- CH), 7.6-8.2 (m, 4H, C6H4NO2) 8.0 (d, 1H, 4-CONH-). e AG Preparation of Exam -Amino(5-ch1orodeoxv-B-D- ribofuganosvl)imidazoleN-[4-nitrophenvl) Slfl-466)) -aminoB-D-ribofuranosylimidazole-N-[(4- (1-343) (0.5g), triphenylphosphine (1.00 g), carbon tetrachloride (0.37 ml), and TH? (25 ml) were combined and stirred at overnight. A methvllcarboxamide (Compound No. nitropheny1)methy1]carboxamide, Compound 23 white added under ambient temperature,under argon, precipitate formed. Dimethylformamide (8 ml) was and the solution was stirred at ambient temperature, The reduced pressure and the resulting oil co-evaporated with (3 x 20 ml). chromatographed on argon, overnight. solvent was evaporated under methanol The resulting viscous oil was silica gel, using 7:1 methylene chloridezmethanol as eluting solvent. The appropriate fractions were combined and concentrated in vacuo to give a yellow foam (0.28 g). The foam was crystallized from cold methanol to give yellow crystals (200 mg), mp = 174- 176°C.

‘H NMR (DMSO-d6) 6 ppm 3.7-3.9 (m, 2H, 5'-CH2), 4.0- 4.4 (m, 3H, 2'-CH, 3'—CH, 4'-cu), 4.5 (d, 2H,-C152-C6!-I4NO2), .4-5.6 (m, 2H, 2'-OH, 3'-OH), 5.6 (d, 1H, 1'-CH), 5.9 (br.s., 2H, 5-NH2), 7.4 (s, 1H, 2-CH), 7.5 - 8.2 (m, 4H, -C6H4NO2), 8.3 (t,lH, 4-CONH-). ribofuranosvl)imidazoleN-I(4-chlorophenv1)methv}1: carboxamide (compound 52 (1-467)) and 5-Amino(5-amino- -deoxy-Q-D-ribofuranosy1)imidazoleN-[(4-chlorophenyl) methy1]carboxamide Hydrochloride (Compound No. 53 (1-468)) dissolved in a mixture of 100 ml DMF, 15 ml acetone and 15 was ml 2,2-dimethoxypropane. Hydrogen chloride gas (approxi- mately 1.0 g) was added and the mixture stirred under argon for 4 hours. The mixture was poured into 50 ml of saturated sodium bicarbonate and evaporated under vacuum at 45°C. ethyl acetate and 25 ml water.

The residue dissolved in a mixture of 100 ml The ethyl acetate phase was separated and washed with 25 ml of water, dried over TLC (silica gel, 9:1 methylene chloride:methanol)showed a significant magnesium sulfate and concentrated to a foam. faster moving impurity in the product which was identified as the 5'-(2-methoxypropane) mixed ketal of the above- identified compound. This was converted to the above- identified compound by dissolving the foam in 100 ml of methanol’ and adjusting the pH to 2.5 with ethanolic hydrogen chloride. After 30 minutes the mixture was neutralized with saturated sodium bicarbonate and concen- This was dissolved in 100 ml of The methylene chloride phase was dried over magnesium sulfate trated to a slurry. methylene chloride, washed with 25 nu. of water. and concentrated to a foam. Drying under vacuum at 40°C for 18 hours yielded 7.2 g (96%) of the above-identified compound.

B. Preparation of 5-Amino(5-iodo-5—deoXv-2.3- isopropylidene-B-D-ribofuranosvljimidazole-4—N-[14- ch1orophenyl)methy1]ca;boxamide A mixture of the product of Step A (25 g, 59 mmole) and methyltriphenoxyphosphonium iodide (76 g, 166 mmole) in 500 ml of methylene chloride was stirred for 30 minutes at room temperature under argon. The resulting solution 150 ml of 5% sodium 100 ml of The solvent was was extracted with 150 ml of water, thiosulfate, water and dried over magnesium sulfate. ml of 1 N sodium hydroxide, removed under vacuum and the resulting oil applied to a 1.31 column of flash grade silica gel prepared in 2:1 hexane:ethyl actetate. The column was eluted with the same solvent to remove impurities then 1:1 hexane:ethy1 the Appropriate fractions’ were combined and evaporated to acetate was used to elute desired product. yield 24.4 g of the above-identified compound as a gummy solid. chromatography to yield an additional 2.3 g of the above- Total yield was 26.7 g (85%).

Impure fractions were again subjected to identified product.

C. Preparation of 5-amino—1-(5-azido-5—deoxv-2,3—O- isopropylidene-B-D-ribofuranosyl)imidazoleN—f(4- chlorophenvllmethvllcarboxamide A mixture of the product of Step B (26.7 g, 50 mmole), lithium azide (14 g, 285 mmole) and 100 mg of 18- crown-6 in 350 ml of DMF was stirred for 8 hours at room temperature under argon. The slurry was concentrated to remove solvent and the residue dissolved in a mixture of 500 ml of ethyl acetate and 100 ml of water. The ethyl acetate phase was separated, washed with water and saturated sodium chloride, and then dried over magnesium sulfate. Evaporation of the solvent yielded 25 g of the above-identified compound as a: yellow gum which still contained solvent. This was used in the next step without further purification. of 5-Amino(5—azidodeoxv-B-D- ribofuranosvl)imidazoleN-I(4-chloroohenvli methyllcarboxamide. 52 (1-467)) The product of Step C, as obtained, was dissolved in D. Preparation (Compound No. ml of 80% trifluoracetic acid and warmed to 50°C for minutes. The solution was evaporated to a syrup at 40°C under vacuum and the residue evaporated twice from 25 ml of water. The syrupy residue was dissolved in 100 ml of ethyl acetate and gently stirred over 100 ml of saturated sodium bicarbonate. Crystaliization began in the ethyl acetate phase and after 1 hour crystals were collected by filtration. These crystals were combined with two additional crops or crystals obtained by concentration of the ethyl acetate phase to yield 15.7 g (77% yield based on the product of Step B). of an analytical sample was 182-183°C.

H NMR (DMSO-d6) 6 ppm, 3.6 (M, 2H, 5'-CH2), 4.0-4.3 3H, 2'-CH, 3'-CH, 4'-CH), 4.3 (d, 2H, -CH2C6H4Cl), 5.4- (m, 2H, 2'-OH, 3'-OH), 5.5 (d, 1H, 1'-CH), 5.9 (br.s., S-NH2), 7.3-7.4 (m, 4H, C6H4Cl), 7.4 (s, 1H, 2-CH), 8.1 H, 4-CONH-). IR (KBr) cm", 2110.

Melting point (my .5 2H, (ti B. Preparation of 5—Amino—1-(5-amino-5—deoxv-B-D- ribofuranosy11imida2o1e—4—N—f(4-chlorophenvll methyl]carboxamide Compound 52 (I-467) in 500 ml of boiling ethanol. (6.5 g, 159 mmole) was dissolved After cooling to 40°C the solution was saturated with argon and 0.5 g of 10% palladium on carbon added. The mixture was stirred under a hydrogen atmosphere for 8 hours. The mixture was saturated with argon and filtered through Celite 505 and concentrated to a syrup which was used in the next step without further purification.

The product of Step E (theoretically 159 mmole) was dissolved in 100 ml of ethanol and 3.5 ml of 6 N hydro- chloric acid added (pH to wet pH paper approximately 3).

(Compound No, 53 The solution was evaporated to a hard syrup. This syrup was dissolved in 50 ml of hot ethanol and diluted with 150 ml of ethyl ether. stirred sealed for 12 hours and the resulting white preci- The resulting gummy precipitate was pitate collected by filtration and washed with ether.

Drying under vacuum at 40°C yielded 6.0 g of the above- identified compound (90% yield based on the compound from Step D).

‘H mm (nuso as) 6 ppm, 3.0-3.2 (m, 23, 5--cnz), 4.0- 4.4 (M, 3H, 2'-CH, 3'-CH, 4'-CH), 4.4 (d, 2H, -CH2-C6H4Cl), .8-6.2 (br., 2H, 2'-OH, 3'-OH), 7.2-7.4 (m, 4H, C6H4C1), 7.8 (S, 1H, 2-CH), 8.3 (br., 3H, NH2'HC1).

Example AI Preparation of S—Amino-l-(5-amino-5—deoxv—B-D- ribofuranosvl)imidazo1e-4—N-(cyclopentyl)narboxamide fiydrochloride ((Compound No. ) 1-270)) This compound was prepared by the same reaction sequence described in Example AH for compound 53 (1-468), (1- 4-N-p-chlorobenzylamide substituting the 4-N-cyclopentylamide, 186), of Table XII for the compound 29 (1-349) of Table XII.

‘H NMR(DMSO—d5) 5 ppm, 1.4-1.9(m, 9H, aliphatic protons), 3.0-3.2 (m, 2H, 5'-CH2), 4.0-4.3(m, 3H, 2'-cu, 3'-cu, 4'-ca), 5.5(d, ‘H, 1'-cm, 5.9(br.s, 2H, 5- NH2), 7.1(d, 1H, 4-CONH-), 7.4(s, 1H, 2-CH). compound 10 cyclopentyl ribofuranosyl)imidazole—4-carboxamide (1-483)) intermediate, jcomnound No.

The ribofuranosyl)imidazolecarboxamide, -amino(5-chlorodeoxy-fi—D- was prepared according to the procedures described in Example AI for 51(1-466), 5-aminofi-D- ribofuranosylimidazolecarboxamide for 5-aminofi-D- r i b o f u r a n o s y 1 i m i d a z o 1 e N - [ ( 4 - nitrophenylmethyl]carboxamide. compound substituting To a 0.1 N sodium methoxide/methanol solution, at 0° under argon, was bubbled methyl mercaptan. To the resulting 0.1 N sodium methylthiolate/methanol solution was added 5—amino(5-chloro-5—deoxyD-ribofuranosyl) imidazo1e—4-carboxamide (0.40 g). The solution was heated of reflux overnight. The solution was cooled and neutral- ized with Dowex so strongly acidic ion exchange resin.

The mixture was filtered and concentrated under reduced pressure. The resulting residue was chromatographed on silica gel, using 4:1 methylene chloride:methanol as the eluting solvent. The appropriate fractions were combined, concentrated under reduced pressure, and vacuum dried to give the above-identified compound as a a ‘white foam (0.28 g).

‘H NMR (ouso-d6) 6 ppm, 2.1(s, an, S-CH3), 3.7- 3.9(m, in, 5'-CH2), 3.9-4.4(m, 3H, 2'-cu, 3'-ca, 4'-CH), .3-5.4 (m, 2H, 2'-OH, 3'-OH), 5.5(d, 1H, 1'-CH), .8(br.s., 2H, 5-NH2), 6.6-6.9(br.m, 2H, 4-CONH2), 7.3(s, H, 2-CH).

Example AK Preparation 5-AminoB—D-ribofuranosylimidazoleN-(4- bromophenv1)carboxamide (Compound No. 55 (1-484)) -Amino—1-(2,3,5—tri-Q-acetyl-E-D-ribofuranosyl) imidazolecarboxylic acid (Srivastava, P.C., et al., J.

Med. Chem. ;1 1207, (1974), (0.75 g) and thionyl chloride (7 ml) were stirred at ambient temperature under a drying tube, evaporated under reduced pressure and the for 15 minutes. The excess thionyl chloride was resulting residue co—evaporated with methylene chloride (3 x 20 ml).

The resulting yellow foam was dissolved in methylene chloride (40 ml) and 4-bromoaniline (0.35 g) was added.

Triethylamine (approximately 0.75 ml) was added until the solution was basic. The solution was stirred at ambient temperature under a drying tube for 2 hours. The solution was washed with water, dried with magnesium sulfate, and concentrated under reduced pressure to give a yellow foam.

The foam was dissolved in methanol (35 ml). A sodium methoxide methanol solution 0.5 N solution) stirred at ambient temperature under a drying tube, for 30 (approximately 0.75 ml of a was added and the resulting solution The solution was neutralized with methanol- The concentrated under reduced minutes. washed Dowex 50 (strongly acidic ion-exchange resin). filtered and pressure to give a pale yellow residue. mixture was The residue was crystallized from methanol (15 ml)/methylene chloride (10 ml) (0.23 g). recrystallized to give off-white crystals (90 mg). 214-216°C (decomp).

‘H NMR (DMSO-d6) 6 ppm, 3.6(m, 2H, 5'-CH2), 3.9-4.3 to give tan crystals The crystals were (m, 3H, 1'-CH, 3'-CH, 4'-CH), 5.2-5.4(m, 3H, 2'-OH, 3'- OH, 5'-Oh), 5.5(d, 1H, 1'-CH), 6.2(br.s., 2H, 5-NH2), 7.4- 7.8 (m, 4H, -C6H4Br), 7.4(s, 1H, 2-CH), 9.5(s, 1H, 4- CONH).

Example AL Preparation of 5-Amino-1—B-D-ribofuranosvl-imidazole—4- N—f(4-bromophenvl)methv1lcarboxamide 0 -487)) compound (Compound No.

This the procedures described in Example J for the 4—p-nitrobenzyl was prepared according to derivative, substituting 4-bromobenzylamine hydrochloride for 4-nitrobenzylamine hydrochloride.

H NMR(DMSO-d6) 6 ppm, 3.5-3.6(m, 2H, 5'-CH2), 3.9- 4.3(m, an, 2'-ca, 3'-cu, 4'-ca), 4.3 (d, 2H, C32-C6H4Br), .1-5.4 (m, 3H, 2'-on, 3--on, 5'-on), 5.5 (d, 1H, 1'-CH), ‘5.9(br.s, 2H, 5-NH2), 7.2-7.5(m, 4H, -C6H4Br), 7.3(s, 1H, -CH), 8.0(t, 1H, 4-CONH-).

Example AM Preparation of S-AminoB-D—ribofuranosy;-imida;o1e—4- E-(4-iodophenyl)carboxamide jcompound No. 57 (1-488)) This compound the procedures described in Example AK for the 4-p-bromophenyl was prepared according to derivative, substituting 4—iodoaniline for 4-bromoaniline.

The final product was recrystallized from ethanol. -229°C H NMR (DMSO-d6) Mp: 6 ppm, 3.5-3.6(m, 2H, 5'-CH2), 3.9-4.4(m, 3H, 2'-CH, 3'-CH, 4'-CH), 5.2-5.4 (m, 3H, 2'- OH, 3'-on, 5'-on), 5.5(d, 1H, 1'-CH), 6.2(br.s., 2H, 5- NR2), 7.4(s, 1H, 2-CH), 7.6-7.7(m, 4H, -C6H4I), 9.5(s, 1H, 4-CONH).

Example AN Preparation of 5-Amino5-D-ribofuranosvlimidazoleN- (4-nitrophenyl)carboxamide (Compound No. 58 (1-482)) This compound was the procedures described in Example AK for the 4-p-bromophenyl prepared according to derivative, substituting 4-nitroaniline for 4- bromoaniline. The final product was recrystallized from methanol to give a yellow powder.

‘H NMR (DMSO-d6) 5 ppm, 3.5-3.6(m, 2H, 5'-CH2), 3.9- .4(m, 3H, 2'-CH, 3'-CH, 4‘-CH), 5.2-5.4 (m, 3H, 2'-OH, 3'-OH, 5‘-OH), 5.6(d, 1H, 1'-CH), 6.4(br.s., 2H, 5-NH2), 7.5(s, 1H, 2-CH), 8.1-8.3 (m, 4H, C6H4NO2), 10.1(5, 1H, 4- coma).

Example 50 Preparation of 5-AminoB-D-ribofuranosv1-imidazo1e N-I2-(4—nitrophenvl)ethvl carboxamidg (1-506)) compound (Compound No.

This to the procedures described in Example J for the 4-p-nitrobenzyl was prepared according derivative, substituting 4-nitrophenethylamine hydrochloride for 4—nitrobenzy1amine hydrochloride.

H lama (DMSO-d‘) 5 ppm, 2.9-3.0(t, 2n, -CH2-C2H‘- N02), 3.4-3.6 (m, 2H, 5'-CH2), 3.9-4.3 (m, 3H, 2'-CH, 3'- CH, 4'-CH), 4.8-5.4(br., 3H, 2'-on, 3'-OH, 5'-on), s.5(d, 1H, 1'-CH), 5.9-6.2(br., 2H, 5-NH2), 7.5-8.2(m, 4H,- C6H4NO2), 7.6(s, 1H, 2-cu), 7.7(t, 1H, 4-CONH).

Example AP Preparation of 5—Amino[1-r4-(4-nitrophenvl)1 piperazinocarbamovllB-D-ribofuranosvlimidazole 60 (1-5081) compound (Compound No.

This according to the procedures described in Example J for the 4-nitrobenzyl derivative, but substituting 1-(4—nitrophenyl)piperazine for 4-nitrobenzylamine hydrochloride. was prepared The product as recrystallized from cold methanol and had a mp of 199- 200°C.

‘H NMR (DMSO-d6) 6 ppm, 3.4-3.6(m, ion; 3'-CH2, piperazohyl methylenes), 3.9-4.3(m, 3H, 2'—CH, 3'-CH, 4'- CH), 5.2-5.4(m, 3H, 2'-OH, 3'-OH, 5'-OH), 5.5(d, 1H, 1'- CH), 6.3 (br.s., 2H, 5-NH2), 7.0—8.l(m, 4n, —C6H4NO2), 7.3(s, 1H, 2-CH).

Example AQ Preparation of 5-Amino(S—deoxv-B-D—ribofuranosvll imidazole-4N~[(4-chlorophenvl)methvl1carboxamide 61 11-509Ll -Amino(5-iodo-5—deoxy-2,3-isopropylidene—fi-D- (Compound No. ribofuranosyl)imidazole—4-N-[(4-chloropheny1)methyl] carboxamide (see procedures described in Example AH for preparation of Compound 53 (1-468), step B) stirred in 30 ml of 50% formic acid overnight. solvent was evaporated under reduced pressure. (0.64 g) was The excess The resulting residue was co-evaporated with water (25 ml) and methanol (25 ml). graphed on silica gel, The resulting yellow foam was chromato- using 9:1 methylene chloride: methanol as eluting solvent. The appropriate fractions were combined and concentrated under reduced pressure to 0.47 g of ribofuranosyl)imidazoleN—[(4—ch1orophenyl)methyl] carboxamide. give 5-amino(5-iodo-5—deoxy-fi-D- -Amino(5-iododeoxy-fi—0-ribofuranosyl) imidazoleN-[(4-chlorophenyl)methyl] carboxamide (0.04 g), palladium on carbon 10% (20 mg), and ethanol (20 ml) were charged to a Parr bottle. The bottle and contents hydrogen. The reaction progress was monitored by HPLC (Waters C18, 55% methanol] 45% 0.1 N acetic acid, 260 nm, 1.0 ml/min). After 24 hour, there was 34% starting material. Fresh catalyst was added (20 mg) and the mixture re-charged with hydrogen (45 p.s.i.). The mixture was shaken for an additional 48 were charged with 45 p.s.i. hours. The reaction mixture contained 30% starting material. The mixture was filtered through Celite, and concentrated under The residue was chromatographed on silica gel, reduced pressure. resulting using ethyl acetate (400 ml) and 5% methanol in ethyl acetate (200 ml) as the eluting solvent. The appropriate fractions were combined and concentrated under reduced pressure to yield 70 mg of a awhite foam. HPLC indicated 9% starting The material was rechromatographed on silica ethyl All fractions containing less than 3% starting material were combined and concentrated under reduced pressure to yield mg of the above—identified compound as a pink foam.

‘H NMR (DMSO-d6) 5 ppm, 1.2-1.3(d, 3!-I, 5'—CH3), 3.7- .3(m, 3H, 2'-CH, 3'-CH2 4'-CH), 4.3(d, 2H, Cfiz-C6H‘Cl), material. gel, using acetate as eluting solvent. .1-5.4(m, an, 2'-on, 3'-on, 1'-CH), 5.8(br.s., 2n, 5- NH2), 7.2-7.4(m, SH, C6H‘C1, 2-CH), 8.1(t, 1H, 4-CONH).

Example AR Preparation of 5-Amino(5-deoxy-S—methvlsulfinvl-B-D- ribofiuragosyl)jmidazole-4—carboxyamide Com ou d No. 62 -510 -Amino(5-deoxymethylthio-fl-D-ribofuranosyl) imidazolecarboxamide (compound 54 (1-483)) of Example AK (0.40 g) peroxide, 30 weight percent, was dissolved in water (20 ml). Hydrogen (0.42 ml), was added and the TLC (6/1, starting solution stirred for 30 minutes. chloride/methanol) methylene indicated some material present. An additional 1.o.ml of hydrogen peroxide was TLC solvent was added and the solution stirred for 15 minutes.

The evaporated under reduced pressure to give a yellow foam. indicated no starting material.

The foam was chromatographed on silica gel, using 3/1, The appropriate fractions were combined and concentrated in methylene chloride/methanol, as eluting solvent. vacuo to give 75 mg of the above-identified compound as a yellow foam.

HPLC C18, 100% ml/minutes, 260 nm) indicated 2 equimolar products. lflo This is consistent with oxidation of the product to a diaster (Waters 0.1 N acetic acid, omeric mixture of sulfoxides.

H NMR (DMSO-d6) 5 ppm, 2.6(s, an, CH3S(O)-), 3.0-3.2 (m, 2H, 5'-CH2), 4.0—4.4(m, 3H, 2'-CH, 3'-CH, 4'- CH) 5.4-5.6(m, 3H, 2'-OH, 3'-OH, 1'-CH), 5.9(br.s., 2H, 5- NH2), 6.6-6.9 (br., 2H, 4-CONH6), 7.3(s, 1H, 2-CH).

Example.AS Preparation of 5-AminoB-D-(5—deoxv-Se methylaminoribofiuranosyl)imidazole—4-carboxamide Com ound No. 63 -517 '-Deoxy-5'-iodo-2',3'-O-isopropylidene-AICA riboside (1.00 g) (ref: P.C. A.R. Newman, T.R.

Srivastava, Mathews, and T.R. Mathews, and R.K. Robins, J. Med. Chem., lg, 1237 (1975)), methylamine 40% weight in water (3 ml), and methanol (30 ml) were combined and heated at reflux for 18 hours.

The reduced pressure.

The reaction gave a mixture of products. solution was cooled and the solvents evaporated under The resulting residue was chromato- graphed on silica gel, using 6/1 methylene chloride] methanol (400 ml) and 3/1 methylene chloride/methanol (300 ml) as the eluting solvent. The fractions containing the slow-eluting component which was desired product were combined and evaporated under reduced pressure to give 0.13 g of 5'—deoxy-5'-methylamino-2',3'-isopropylidene- AICA riboside. '-deoxy-5'-methylamine-2',3‘-isopropylidene AICA riboside (0.13 g) was heated at 60°C in 75% formic acid (20 ml) for 1.5 hour. solvent evaporated under reduced pressure to yield a white The solution was cooled and the foam. The foam was dissolved in water (5 ml) and applied to a short column of Dowex 50 strongly acidic ion-exchange The column was washed with water then eluted with 1 M NH4OH in 20% methanol/water. resin.

The solvent was evapor- ated under reduced pressure and the resulting residue co- evaporated with methanol (3 x 20 ml) to yield 75 mg of the above-identified product as an off-white foam.

H NMR (D6-DMSO-d6) 5 ppm, 2.3(s, an, CH3N), 2.5- 2.7 (m, 2H, 5'-CH2), 3.3-3.4(br., 1H, MENH), 3.9-4.3(m, 3H, 2'- CH, 3"CH, 4"CH), 5.1-5.4(m, 2H, 2'-OH, 3"OH), 5.4(d, 1H’ 1"-CH), 6o2(broso, 2H, 5— 6u6—6n8 (bro, 2H’ 4- CONH), 7.2(S, 1H, 2'CH).

Exam 9 A Preparation of 5-Amino—1-B-D-ribofuranosvlimidazoleN- (2-chlorophenyl)carboxamide (Compound No. 64 (1-519)) This compound was prepared according to the proce- dures decribed in Examples AK for compound 55 (1-484) for the substituting 2- —p-bromophenyl derivative, chloraniline for 4-bromaniline. The final product was recrystallized from methylene chloride (20 ml)/methanol (1 ml) to yield 0.25 g of the above-identified product. Hp = 131-135°C.

‘H NMR (DMS0-d6) 6 ppm, 3.5-3.6(m, 2H, 5'-CH2), 3.9 - 4.3(m, 3H, 2'-CH, 3'-CH, 4'-CH), 5.2-5.4(m, 3H, 2'-OH, 3'-OH, 5'-OH), 5.5(d, 1H, 1'-CH), 6.2(br.s., 2H, 5-NH2), .0-8.4 (m, 5H, C6H4Br, 2'-CH), 9.1(S, 1H, 4-CONE).

Example AU Preparation of 5-Amino5-D-(5-benzv1amino deoxyribofuganosyl)imidazolecarboxamide (Compound No. 66(1-531)) Srivastava, and methanol (40 ml) were combined and heated at reflux for 24 hours. Then, the procedures (1-517) followed to give the above-identified compound.

H NMR (DMSO-d6) 6 ppm, 2.7 (d, 2H, -CH2-CSHS), 3.3- 3.4(br., 1H, -NH —CH2C6H5), 3.9-4.3(m, 3H, 2'-CH, 3'-ca, 4'-CH), 5.1-5.4(m, 2H, 2'-OH, 3'-OH), 5.4(d, 1H, 1-CH), 6.1(br.s., 2H, 5-NR2), 6.6-6.8(br., 2H, 4-CONH2), 7.2- .4(m, en, -CGHS, 2-ca). described in Example AS for Compound 63 were Exam e V Preparation of 5-AminothioB-D deoxyribofuranosyl)imidazolecarboxamide 5 Com ound No. 67 -535 A. Preparation of 5'-Deoxy-2'.3'—isooroDv1idene bromo-AICA Riboside To a solution of 5'—deoxy-2',3‘-isopropy1idene—AICA riboside (2.90 g) P.C. Srivastava, A.R.

T.R. Robins, J. Med. Chem., (ref: and R.K.

Newman, (1975)) in chloroform (100 ml), bromosuccinimide in small portions over 20 minutes. added N- The ambient temperature for 30 solution was stirred at minutes. The solution was washed with water, twice with brine, and then dried over magnesium sulfate. The solvent was evaporated in vacuo to yield a dark foam. The foam was passed through a column of silca gel, eluting with 9:1 methylene chloride:methanol. The fractions containing product were combined and concentrated under reduced pressure to yield 2.02 g of reddish—brown foam.

B. Preparation of 5'-Deoxv-2'.3'isopropvlidene thio AICA Riboside Postassium sulfate (3.7 g) was heated at reflux in ethanol (20 ml) for 15 minutes.

To the filtrate was added 5'-deoxy-2',3'-isopropylidene- 2-bromo AICA riboside (from step A). The mixture was heated at 100°C in a steel bomb for 5.5 hours. The mix- The pH of the filtrate was adjusted to about 5-6 with acetic acid, and the solvent The mixture was filtered. ture was cooled and filtered. evaporated under reduced pressure. The resulting residue was passed through a column of silica gel, eluting with 7/1, methylene chloride/methanol. The fractions contain- ing the product were combined and concentrated under The foam was stirred in methylene chloride (50 ml), then filtered to reduced pressure to give a dark brown foam. yield a pale purple powder. The powder was stirred in cold methanol, then filtered and vacuum dried to yield .52 g of a pale yellow solid. Mp = 211-214 (decomposition).

C. Preparation of 5-Aminothio(deoxv-fi-D- gibofuranosyl)imidazole—4-carboxamide (Compound 67 (1-535)) ’-deoxy-2',3'-isopropylidenethiol AICA riboside (0.45 g) (from step 8) was stirred in 50% formic acid (30 ml) at 50°C for 1 hour. The solvent was evaporated under “overnight. reduced pressure. The resulting residue was co—evaporated with methanol (2 x 20 ml). (25 ml), then stirred at room temperature The mixture was filtered and the filtrate concentrated under reduced pressure to yield a greenish The resulting solid was warmed in methanol foam. The foam was chromatographed on silica gel, using /1, methylene chloride/methanol, as the eluting solvent.

The appropriate fractions were combined and concentrated under reduced pressure to give a yellow foam. The foam was crystallized from cold methanol to yield 69 mg. of the above-identified compound mp = 201-203°C, (decomposition).

‘H NMR (DMSO-d6) 6 ppm l.3(d, 3H, 5'-CH3), 3.6-4.5(m, 3H, 2'-CH, 3'-CH, 4'-CH), 5.0-5.2 (m, 2H, 2'-OH, 3'-OH), .6(br.s., 2H, 5-NH2), 6.0(d, 1H, 1‘-CH), 7.0(br., 2H, 4- COEH), 12.0 (br.s., 1H, -SH).

Example AW Preparation of N,N'-bis-(5-aminoQ-D-ribofuranosyl imidazolecarbonyl)-1,6—diaminohexane (Compound No. 68 (1-538)) N-succinimidylamino-l-(2,3,5-tri-Q-acetyl-fi-D- ribofuranosyl-imidazole—4-carboxylate (2.50 g) (ref: P.C., et al., J. Med. Chem. ;1:l207 (1974)), 1,6-hexane diamine (0.300 g), triethylamine (0.5 ml), and methylene chloride (35 ml) Srivastava, were combined and stirred at room temperature for 18 hours. The title compound was prepared according to the procedures described in Example J. The final product was crystallized from methanol to yield 0.32 g of the above-identified compound. Mp - 181- 185°C.

H NMR data reported as for half the symmetrical dimer. ‘H NMR (DMSO-d6) 6 ppm, 1.2-1.5(m, 4H, 3 and 5 methylenes of N-hexyldicarboxamide), 3.0-3.2(m, 2H, a methylene of N—hexyl dicarboxamide), 3.5-3.6(m, 2H, 5'- CH2), 3.8-4.3(m, 3H, 2'-H, 3'-cH,'4'-CH), 5.l—S.4(m, 3H, 2'-OH, 3'-OH, 5'-OH), 5.5(d, 1H, 1'-CH), 5.9(br.s., 2H, 5- NR2), 7.3(s, 1H, 2-Ch), 7.4 (t, 1H, 4-CONH). ribofuranosvlimidazolecarbonyl)-1.4-diaminocvclohexane (Compound No. 69 (1-549)) This compound was prepared according to the proce- dures described in Example AW for compound 68 (1-538), substituting 1,4—diaminocyc1ohexane for 1,6- hexanediamine.

H NMR data are reported as for half the symmetrical 1H NMR (DMS0-d6) 6 ppm 1.3-1;8(m, 4H, cyclohexane 3.5-3.7(m, 3H, 5'-CH2, methine), 3.8-4.3(m, 3H, 2'-CH, 3'-CH, 4'-CH), 5.1-5.4(m, 3H, 2'-OH, 3'-OH, 5'-OH), 5.5(d, 1H, 1'-CH), 5.9(br.s., 2H, 5-NH2), 7.1(d, 1H, 4-CONH) 7.3(s, 1H, 2-CH). dimer. methylene protons), cyclohexane Example AY Preparation of 5-Aminothio(5-aminodeoxv-B-D- ribofuranosvl) imidazolecarboxamidg jcompound No. 70(1-551)) A. Preparation of 5-Deoxv-5'—iodobromo-2'.-3'- isopropylidene AICA Riboside 2—Bromo-2'3'-isopropylidene AICA riboside S.Suzaki, A. (1976) and methylene chloride (4.50 g) (ref: Chem.

Bull. iodide T.Miyoshi, 29, g:2089, methyltriphenoxyphosphonium (16.2 g), (125 ml) combined and stirred at room temperature for 16 hours.

The mixture was washed with water, 0.5 M NAOH (100 ml), 5% NaS2O3 (150 ml), The pressure to give an orange oil. cold diethylether. give 3.53 g of a grey powder.

Yamazaki, Pharm. were and brine, then dried over magnesium sulfate. solvent was evaporated under reduced The oil was triturated in The resulting mixture was filtered to The mother liquor was concentrated under reduced pressure to give an orange oil.

The oil was applied to a short column of silica gel. The column was washed with methylene chloride, then the product eluted with 9/1, methylene chloride/methanol (2 ml). The appropriate fractions were combined and concen- trated under reduced pressure to give an orange tar. The tar was triturated with cold diethyl ether. The mixture was filtered to yield an additional 0.94 g of a gray powder. The combined powder (4.47 g) was chromatographed on silica gel, using 2/1, ethylacetate/hexane, as eluting solvent. The appropriate fractions were combined and concentrated under reduced pressure to yield a yellow foam (4.02 g).

B. Preparation of 5'—Azido-5'deoxybromo-2‘,3'- isopropylidene AICA Riboside '-deoxy-5'-iodobromo-2',3‘-isopropylidene AICA riboside (4.02 g) lithium azide (1.82 g), and DMF (65 ml) were combined and stirred at ambient temperature for 2 hours. The solvent was evaporated under reduced pressure to give a yellow oil. The oil was dissolved in ethyl acetate (200 ml), washed with water and brine, then dried over magnesium sulfate. The solvent was evaporated under reduced pressure to give a yellow foam (3.01 g).

C. Preparation of 5'-Amino—5'-deoxvbromo-2'.3‘- isopropylidene AICA Riboside '—azido-5'—deoxy-2—bromo-2‘,3‘-isopropylidene AICA riboside (2.00 g), and TH? (100 g) were combined and stirred at room temperature for Concentrated NH40H (15 ml) was added and the solution heated at reflux for 6 hours. triphenylphosphine (1.83 g), hours.

The solution was cooled and the solvent evaporated under reduced pressure.

The resulting residue was coevaporated with methanol (2 x ml). cold methanol (25 ml) for 30 minutes. The mixture was filtered to give an off-white powder.

The resulting residue was stirred i: The solid was recrystallized from methanol to give a white powder (0.73 g).

Q-gibofiuganosyl)imidazolecarboxgmige{CompoundNo, Potassium sulfide (1.0 g) was heated at reflux in ethanol (10 ml) for 15 minutes. and to the filtrate was added 5'-amino-5'-deoxybromo- The mixture was filtered The mixture The The filtrate was again ',3'-isopropylidene AICA riboside (0.50 g). was heated in a steel bomb at 110°C for 5 hours. mixture was cooled and filtered. filtered, then concentrated under reduced pressure to give a yellow tar. 3/1. solvent.

The tar was chromatographed on silica gel, eluting The appropriate fractions were combined and using methylene chloride/methanol, as concentrated under reduced pressure to give a yellow glass (0.12 g). acetic acid (8 ml) and stirred at room temperature for 1 The glass was dissolved in 80% of trif1uoro- hour. The solvent was evaporated under reduced pressure to give a yellow solid. The solid was stirred in diethylether/ethanol (10 ml of 95/5), then filtered and dried to yield a yellow solid (55 mg).

‘H NMR (DMSO-d6 + D20) 6 ppm, 2.6-2.9(m, 2H, '-CH2-), 3.8-4.5(m, 3H, 2'-CH, 3' CH, 4'-CH), 6.2(d, 1H, 1'-CH).

Example 5; Preparation of 5-Amino15-azido —5-deoxv-B-D- ribofuranosvllimidazole N-I14-nitrophenv1)- eth carboxam'de (Compound No. 7; (1-562)) This compound was prepared according to the procedures described in example AH for compound 52 (1- ), derivative), substituting compound 23 (1-343) (1-349) (p-nitrobenzyl for compound 29 (p-chlorobenzyl derivative).

‘H NMR (nnso-as) 5 ppm, 3.5-3.7(m, 2H, 5'-CH2), 3.9 -4.4(m, 3H, 2'-CH, 3'-CH, 4'-CH), 4.4-4.5(d, 2H, -CH2- PhNO2), 5.4-5.5(m, 2H, 2'-OH, 3'-OH), 5.5(d, 1H, 1'-CH), .9(br.s., 2H, 5-NH2), 7.4(s, 1H, 2-CH), 6.5-8.2 (m, 4H, -C6H‘NO2), 8.3(4, 1H, 4-CONH-).

Example 55 Preparation of 5-Amino(5-aminodeoxv-B-D- ribofuranosvl)imidazoleN114-nitrophenvl) methyl]carboxamide This compound was according to the procedures described in Example AH for compared 53 (1- 468), substituting the p-nitrobenzyl amide _derivative (compound 23 (1-343)) the derivative (compound 29 (1-349)).

‘H NMR (nmso + D20) 6 ppm 2.6-2.8(m, 2H, 5'-ca,-), 3.8-4.3(m, 3H, 2'-CH, 3'-CH, 4'-CH), 4.4-4.5(m, 2H, -CH2- C6H‘NO2), 5.4(d, 1H, 1'-CH), 7.3(s, 1H, 2-CH), 7.5—8.3(m, SH, CH2C6H‘NO2, 4-CONH). prepared for p-chlorobenzyl amide N-[(4-(trifluoromethylphenyl)methy1]ca;boxamide (Compound No.

This the procedures described in Example J for the p—nitrobenzy1 -572)) compound was prepared according to derivative substituting 4-(trifluoromethyl)benzylaminezfor 4-nitrobenzyl amine hydrochloride. The final product was recrystallized from methylene chloride/methanol. Mp = 137 - 140.

H NMR (DMSO-d6) 6 ppm 3.5 - 3.7 (m, 2H, 5'-CH2), 3.9 - 4.4 (m, 3H, 2'-CH, 3'-CH, 4'-CH), 4.4 - 4.5 (d, 2H, - CH2-PhCF3), 5.2 - 5.5 (m, 3H, 2'-OH, 3'-OH, 5'-OH), 5.5 (d, 1H, 1'-CH), 5.9 (br.s., 2H, 5-NH2), 7.3 (S, 1H, 2-CH), 7.4 - 7.7 (m, 4H, -C6H4CF3), 8.2 (t, 1H, 4-CONH).

Exam -sulfa o hen 1 met carbo am'de '1Compound No. 75 11-577)) This compound was prepared according to the procedures described in Example J for the p-nitrobenzyl derivative, substituting 4-(aminomethyl)benzene sulfona- mide hydrochloride for 4-nitrobenzylamine hydrochloride.

‘H NMR (DMSO-d6) 6 ppm, 3.5-3.7(m, 2H, 5'-CH2-), 3.9- .4(m, 3H, 2'-ca, 3'-CH, 4'-CH), 4.4-4;5(d, 2H, -CH2- C6H4SO2), 5.2-5.4(m, 3H, 2'—OH, 3'-OH, 5'-oH),~s.5(d, 1H, 1‘-CH), 6.0(br.s., 2H, 5-NH2), 7.3(br.s., 2a, —so2Nn2), .4(s, 1H, 2—CH), 7.4-7.8(m, 4H, -C6H4-), 8.2 (t, 1H, 4- CONH-).

Example BD Preparation of 5-Amino(5-(4—chlorobenzv1—amino) deoxvfi-D-ribofuranosvl)imidazolecarboxamide 76 (1-578)) '-amino-5'-deoxy-AICA-riboside (0.50 g) (compound No. 21 (1-227)) of Table VIII, 4-chlorobenzyl iodide (0.50 g), potassium carbonate (0.26 g), and DMF (15 ml) were combined and stirred at room temperature for 16 hours. lcompound No.

The solvent was evaporated under reduced pressure and the The insolubles were removed by filtration and the filtrate concentrated under reduced pressure. The residue was chromatrographed on silica gel, using 3:1, methylene chloridezmethanol, The fractions containing the slower moving of the two products resulting residue stirred in warm ethanol (35 ml). resulting as eluting solvent. were combined and concentrated under reduced pressure to yield a tan foam (0.21 g) H NMR (ouso-as + D2) 6 ppm 2.9-3.0 (m, 2H, 5'-CH2- ), 3.9(s, 2H, -CH2-CGH‘), 3.9-4.3(m, 3H, 2'-CH, 3'-CH, 4'- CH), 5.5(d, 1H, 1‘-CH), 7.3(s, 1H, 2-CH), 7.4(m, 4H, -C6H4Cl). xa e B -Amino 5—deox - imidazole; (compound No. 77 (1-588)) '-deoxy AICA (1.00 g) (ref: P.C. srivastava, A.R. Newman, T.R. Mathews, and R.F. Robins, J. potassium hydroxide (4.0 ml) for 5 hours.

Pre aration of D-ribofuranos - riboside was heated at reflux in N The solvent was evaporated under reduced pressure and the resulting (4 x 10 ml). The resulting residue was diluted with ethanol (15 ml) and a residue co~evaporated with ethanol fine precipitate was filtered. Upon setting for several The and the combined solid days, the filtrate gave an additional precipitate. microscopic solid was collected, material was dissolved in water (20 ml) and neutralized The solvent was evaporated under reduced pressure to give a with Dowex sow strongly acidic ion exchange resin. dark tar. The tar was dissolved in 80% acetic acid (20 ml) and gently heated (60°C). under reduced presure to give a dark tar. (2 x 15 ml). residue was chromatographed on silica gel, The solvent was evaporated The tar was co- evaporated with methanol The resulting using 3/1, The appropriate fractions were combined and concentrated under methylene chloride/methanol, as eluting solvent.

The tar was co- evaporated with tolune (3 x 20 ml), then vacuum dried to yield a dark brown, hygroscopic foam (110 mg).

‘H NMR (D2) 6 ppm, 1.3(d, 3H, 5'-CH3), 4.0-4.5(m, 3H, reduced pressure to yield a dark tar.

'¥CH, 3'-CH, 4'-CH), 5.6(d, 1H, 1'-CH), 6.4(s, 1H, 4- CH), 7.7(s, 1H, 2-CH).

Example BF re aration of 5-Amino—1- 5-deox dieth laminoribo- fiuranosyl)imidazole-4—carboxamide (Compound No.65 (1-522) '-deoxy-5'-iodo-2',3'—isopropylidene AICA riboside (1.00 g) P.C. Srivastava, A.R. Newman, T.R.

Mathews, and R.K. Robins, J. Med. Chem. lg; 1237, (1975)), (ref.: diethylamine (2.5 ml of 40 wt% in water), and methanol (30 The procedures described in Example AS for compound 63 (1- ml) were combined and heated at reflux for 18 hours. ) were followed to give the above-identified compound.

‘H NMR (DMSO-d6) 6 ppm 0.9 (t, en, methyl groups on '-diethylamine), 2.4-2.7 (m, 6H, 5'—CH2, methylene groups on 5'-diethylamine), 3.3-4.2 (m, 3H, 2'-CH, 3'-CH, 4'- CH), 5.2 (br., 2H, 2'-on, 3'-on), 5.4(d, 1H, 1'-ca), 5.9 (br.s., 2H, 5-NR2), 5.7-5.9 (br., 2H, 4-conaz), 7.3(s, 1H, 2-CH). [3gitrophenyl)propyl]carboxamide (Compound No.

This the procedures described in Example J for the p-nitrophenyl (1-566)) compound was prepared according to derivative, substituting 3-(4-nitrophenyl)propylamine (ref: G.W. Hardy, et al., J. Med. Chem. ;_: 1108, (1989)) for p—nitrobenzylamine hydrochloride.

H NMR (DMSO-d6) 6 ppm 1.7-3.2 (m, 6H, - CH2 CH2-), 3.5-3.6 (m, 2H, 5'-CH2), 3.9-4.3 (m, 3H, 2'-ca, 3'-CH, 4'- cu), 5.2-5.4(m, 3H, 2'-on, 3'-OH, 5'—OH), 5.5(d, 2H, 1'- CH), 5.9(br.s., 2H, 5-NH2), 7.3 (5, 1H, 2-CH), 7.5-8.2 (m, H, -CH6H4NO2, 4-CONH-). 'midazo e - 4-ch o o hen -D-ribo uranos carboxamide Com ound No. 78 1- A. Preparation of 5-amino—1-(5-azido—5-deoxv-2.3-di-O- acetvl-B-D-ribofuranosvl)imidazoleN-I4- chlorophenyl)methyl]carboxamide Compound 52 (example AH), 2.4 g (5.8 mmol), dissolved in a mixture of 20 ml of diemthylformamide and was ml of pyridine. The solution was cooled to 3°C under argon, and acetic anhydride, 1.5 g, (14 mmol), was added.

The mixture was allowed to warm to room temperature over 18 hours and then concentrated to a syrup. The syrup was dissolved in 25 ml of methylene chloride and washed with 3 x 15 ml of water, dried over magnesium sulfate and mixture was stirred under a hydrogen atmosphere for 30 of ethanol and 50 mg of 10% Pd on carbon was added. minutes, filtered and the filtrate evaporated to yield 300 mg of the desired product as a white foam.

‘H mm (DMSO-d6) 6 2.0 (s, 3H, cnsco-), 2.1 (s, 3H, CH3CO-), 2.9 (m, 2H, 5'-CH2), 4.1 (m, 1H, 4'-CH), 3.4 (br. s, 2H, 5'-NH2) 4.4 (d, 2H, -CH2-CGH‘-Cl), 5.3 (m, 1H, 3'- CH) 5.6 (m, 1H, 3'-CH), 5.8 (d, 1H, 1‘-CH), 6.4 (br. 5, 23-1, 5-NR2), 7.3 (m, 4H, -can‘-c1), 7.4 (s, 1H, 2-cn), 8.1 (t, 1H, 4-CONH-).

Claims (51)

CLAIMS:

1. An AICA riboside analog of formula: N 1 R1 RI n,o on, and pharmaceutically acceptable salts thereof wherein X is —O~ or~CH2~; R1 is hydrogen, amino, hydrocarbylamino, acylamino or dihydrocarbylaminoalkyleneimino; R2 is hydrogen, cyano, hydrocarbylimidate, carboxamidoxime, hydrocarbyloxyamidine, carboxamide, or carboxylic acid or an amide, ester, thioester or salt thereof; R3 is hydrogen, hydrocarbyl, amino, hydrocarbylamino, halogen, hydroxy, hydrocarbyloxy, sulfhydryl, or hydrocarbylthio; R4 and R5 are independently hydrogen, hydrocarbyl, acyl or hydrocarbyloxycarbonyl; R6 is hydrogen, hydrocarbyl, halogen, hydroxy, hydrocarbyloxy, sulfhydryl, hydrocarbylithio, sulfamyloxy, amino, hydrocarbylamino,. azido, acyloxy or hydrocarbyloxycarboxy or phosphate ester group; provided that when R, is amino, R2 is unsubstituted carboxamide, R3 is hydrogen; R, and R5 are hydrogen, acyl or hydrocarbyloxycarbonyl; then R6 is not hydroxy, acyloxy or hydrocarbyloxycarhoxy; or a pharmaceutically acceptable salt thereof for use in a method of reducing or preventing tissue damage associated with undesired decreased blood flow.

2. A compound according to claim 1 wherein the tissue is cardiac muscle,-brain, skin tissue, placenta, liver, pancreas, kidney, or a tissue of the gastrointestinal system.

3. A compound according to claim 1 or 2 wherein the damage is associated with congestive heart failure, or is due to cardiopulmonary bypass, or organ transplant or cardio-pulmonary arrest or is associated with vascular disease.

4. A compound according to any one of the preceding claims claims wherein the analog is administered as a prophylatic.

5. An AICA riboside compound or salt thereof as defined in claim 1 for use in a method of treating an animal having an undesired region of decreased blood flow, which method comprises the administration to said region of said compound.

6. A compound according to claim 5 wherein cells in and about the region of decreased blood flow are undergoing net adenosine triphosphate catabolism due to a pathologic process.

7. A compound according to claim 5 or 6 wherein said undesired region of decreased blood flow is caused by or believed to be caused by coronary artery occlusion, myocardial infarct, vascular thrombosis, atherosclerosis; or is associated with angina pectoris, transient ischemic attack, cardioplegia, organ transplant or reconstructive surgery: or causes or is believed to cause myocardial arrhythmia or a stroke; or is caused by platelet or granulocyte aggregation or occulusion, vascular spasm, an embolus, inflammation or vasculitis.

8. A compound as defined in claim 1 for use in a method of enhancing the extracellular concentration of adenosine around cells having a decreased ratio of synthesis of adenosine triphosphate to breakdown of adenosine triphosphate due to a pathologic process. 120

9. A compound according to claim 8 for administration as a prophylactic.

‘l0. A compound according to claim 8 wherein the pathologic process is a heart attack, a seizure, a stroke, an inflammatory disease, an autoimmune disease, peripheral ‘vascular disease, transient ischemic attack, coronary artery occlusion or myocardial infarct.

11. A compound according to claim 10 wherein the pathologic process is angina pectoris.

12. A compound as defined in claim 1 for use in a method of reducing or preventing tissue damage in an animal associated with a pathologic process.

13. A compound according to claim 12 wherein the pathologic process is chronic obstructive pulmonary disease, arthritis, an autoimmune disease, sepsis, burns, hyperoxia, inflammatory bowel disease, dialysis, aspiration, adult respiratory distress syndrome, chronic cystitis, inflammation cardiopulmonary arrest, or infection. 121

14. A compound as defined in claim 1 for use in a method of treating a patient having chronic low adenosine levels or who would benefit from increased systemic or central nervous system adenosine levels.

15. A compound according to claim 14 wherein the patient has irritable bowel syndrome, insomnia, autism, schizophrenia, anxiety, cerebral palsy or other neuropsychiatric symptoms.

16- A compound as defined in claim 1 for use in a method of reducing or preventing neural tissue damage caused by a neurodegenerative disease or by excitotoxicity due to increased release of excitatory amino acids in an affected animal.

17. A compound according to claim 16 wherein the neural tissue is brain or spinal chord.

18. A compound according to claim 16 or 17 wherein the excitotoxicity is caused by or causes brain trauma, or is caused by or causes a neurodegenerative condition.

19. A compound according to claim 18 wherein the neurodegenerative condition is Parkinson's Disease, Alzheimer's Disease, Amyotrophic Lateral Sclerosis or Huntington's Disease.

20. A compound as defined in claim 1 for use in method of inhibiting epileptic seizures in an animal.

21. A compound as defined in claim 1 for use in method for treating allergic conditions in an animal.

22. A compound according to claim 21 wherein the allergic condition is associated with asthma or hayfever.

23. A compound according to any one of the preceding claims enhancing the use in a method which comprises the administration of a compound which inhibits adenosine transport at a therapeutic dose which does not substantially affect blood pressure or heart rate or does not cause coronary steal.

24. A compound according to any one of the preceding claims wherein Riis amino,I§ is carboxamide wherein one of the amide hydrogens (attached to the nitrogen atom) is benzyl optionally substituted with 1 to 3 substituents indepdently selected.from halogen, alkyl, aryl, nitro, amino, hydrocarbylamino, sulfhydryl, hydrocarbylthio. hydroxy, hydrocarbyloxy, trifluoromethyl or sulfonamido7 R3 is hydrogen, R4 and R3 are hydrogen, and R6 is amino; or pharmaceutically acceptable salts thereof-

25. A compound according to claim 24 wherein R2 is N-(4—chlorobenzyl)carboxamide. 123

26. A substituted-imidazole AICA riboside analog of formula: R50 OR‘ wherein X is or —CH2-, R1 is amino, hydrocarbylamino, or dihydrocarbylaminoalkyleneimino; R2 is carboxamide wherein one of the amide hydrogens nitrogen (attached to the replaced by alkyl, cycloalkyl, or aryl or aralkyl optionally substituted with 1 to 3 substituents independently selected from halogen, atom) is optionally alkyl, aryl, nitro, amino, hydrocarbylamino, sulfhydryl, hydrocarby1thio,rwdroxy,hydrocarbyloxy,trifluoromethyl, or sulfonamide, R2 is carboxamide wherein both amide hydrogens are replaced by alkyl or together by an alkylene or aralkylene group to form a ring; R: is —C(O)—S-R7 alkyl, substituted wherein R7 is cycloalkyl, or aryl or aralkyl optionally with 1 to 3 substituents independently selected from halogen, alkyl, aryl, nitro, amino, hydrocarbylamino, sulfhydryl, hydrocarbylthio, hydroxy, hydrocarbyloxy, trifluoromethyl, or sulfonamido; R3 is hYdf°9€n. hydrocarbyl, amino, hydrocarbylamino I halogen, hydroxy, hydrocarbylthio; R hydrocarbyloxy, and RS sulfhydryl, or 4 are independently hydrogen, hydrocarbyl (of 1 to about 18 carbon atoms), hYd|'0XY, ha'08€n, hydrocarbyloxy, sulfhydryl, hydrocarbylthio, acyl or 124 sulfamyloxy, amino, hydrocarbylamino, azido, acyloxy, hydrocarbyloxycarboxy or phosphate ester or salt thereof; provided that when X is or —CH2-, R1 is amino, R3 is unsubstituted carboxamide, R3 is hydrogen, R‘ and R5 are independently hydrogen, acyl or hydrocarbyloxycarbonyl, then R6 is not hydrogen, hydroxy, acyloxy or hydrocarbyloxycarboxy or when.both R4 and.Rs are hydrogen, R6 is not a phosphate ester; and provided that when X is oxygen, R1 R2 is unsubstituted carboxamide, R3 is sulfhydryl, and R4 and R5 are both hydrogen, then R6 is not acetoxy; when X is oxygen, R1 is is amino, amino, R2 is ‘unsubstituted carboxamide, R3 is chloro, bromo, amino or methoxy, and R‘ and R5 are both hydrogen, then R6 is not hydroxy, or, when R4 and R5 are both acetyl, then R6 is not acetoxy; and provided further that when X is oxygen, R1 is amino, R: is henzylcarboxamide or pr iodophenylcarboxamide, and R3 is hydrogen, then R‘ and R5 are not both hydrogen, and R65£;not_hydroxy; or when R: is p—iodophenylcarboxamide, thenig andrg are not both acetyl and R6 is not acetoxy; and provided that when R? is carboxamide substituted by cycloalkyl, the cycloalkyl group is not 1—adamanty1; thereof.

27. A compound according to any one of the preceding claims wherein R6 is amino or hydrocarbylamino.

28. A compound according to claim 24 wherein R6 is amino- and_pharmaceutically acceptable salts 125

29; A compound according to any one of claims 26 to 28 wherein R6 is benzylamino optionally substituted with 1 to 3 substituents independently selected from halogen, alkyl of 1 to 8 carbon atoms, nitro, alkylamino, alkylthio or alkoxy.

30; A compound according to any one of the preceding Claims: wherein R115 amino, R2 is unsubstituted carboxamide and R3, R, and R5 are hydrogen.

31. A compound according to any one of the preceding claims wherein R1 is amino and R3 is hydrogen.

32. A compound according to claim 31 wherein R2 is carboxamide wherein an amide hydrogen is replaced by aralkyl optionally substituted with l to 3 substituents independently selected from halogen, alkyl, aryl, nitro, amino, hydrocarbylamino, sulfhydryl, hydrocarbylthio, hydroxy, hydrocarbyloxy, trifluoromethyl or sulfonamido.

33. A compound according to claim 32 wherein R6 is hydroxy, azido or amino.

34. A compound according to claim 33 wherein R2 has an amide hydrogen replaced by a para-substituted benzyl group. 126

35. A compound according to claim 34 wherein R2 is N44—ch1orobenzy1)carboxamide, N—(4-nitrobenzyl)carboxamide or N-(2,4-dichlorobenzyl)—carboxamide.

36. A compound according to claim 35 wherein R6 is amino or azido-

37. A compound according to claim 36 wherein Fg and R5 are hydrogen and X is oxygen.

38. A compound according to claim 36 wherein R, and F5 are independently alkyl, acyl or hydrocarbyloxycarbonyl.

39. A compound according to claim 38 wherein R; and Fg are acetyl and X is oxygen.

40. A compound according to claim 37 or 39 which comprises a hydrochloride salt, or a salt selected from hydrobromide, hydrosulfate, sulfate, hydrophosphate or oxalate.

41. A compound according to claim 35 wherein R; and R5 are hydrogen, R6 is hydroxy and X is oxygen.

42- A compound according to claim 31 wherein R5 is N-(cyclopropyl)carboxamide, N—(cyclopentyl)carboxamide or —c(o) -—S-R7. 20 127

43, An AICA riboside analog according to claim 26 wherein R1 is amino, F5 is carboxamide wherein one of the amide hydrogens is replaced by a substituted or unsubstituted hydrocarbyl group, Fg is hydrogen, R‘ and R5 are hydrogen or hydrocarbyloxycarbonyl, and R6 is hydroxy or amino.

44. An AICA riboside analog according to claim 43 wherein R2 is an aralkyl group.

45. An AICA riboside analog according to claim 26 wherein R1 is an amino, R? is carboxamide, R3 is halogen or sulfhydryl, R; is H, Fg is H and R6 is hydroxy.

46. An AICA riboside analog according to claim 26 wherein R1 is amino, R2 is carboxamide; R3,.R4 and R5 are hydrogen and R5 is amino.

47. An AICA riboside analog according to claim 26 wherein R1 is amino, R2 is carboxamide, R3 is hydrogen, R4 is alkyl, F3 is hydrogen and R6 is hydroxy. 10 128

48_ A compound according to claim 26 wherein X is or —CH2—, R1 is amino, hydrocarbylamino, or dihydrocarbylaminoalkyleneimino; R2 is piperazinocarbamoyl optionally substituted with a hydrocarbyl group optionally substituted with 1 to 3 substituents independently selected from halogen, alkyl, aryl, nitro, amino, hydrocarbylamino, sulfhydryl, hydrocarbylthio. hydroxy, hydrocarbyloxy, trifluoromethyl or sulfonamidoi R5 is hydrogen, hydrocarbyl, amino, hydrocarbylamino, halogen, hydroxy, hydrocarbyloxy, sulfhydryl, or'hydrocarbylthio;Ig and.Rs are independently hydrogen,-hydrocarbyl (of 1 to 18 carbon atoms), acyl or hydrocarbyloxycarbonyl; and R6 is hydrogen, hydrocarbyl, hydroxy, sulfhydryl, hydrocarby1thio,.amino, hydrocarbylamino, azido, acyloxy, hydrocarhyloxycarboxy or phosphate ester or salt thereof. hydrocarbyloxy, 129

49. A compound according to claim 26 selected from: 5-amino[3-D-ribofuranosylimidazoleN-(cyc|opentyl)car’ooxamide. 5-amino-1 -[3-D-ribofuranosylimidazoleN-(cyclopropyncarboxamide. 5-amino-5'-sulfamoyl(5-D-ribofuranosylimidazolecarboxamide. 5-amino-1 -(2-O-methyl-&D-ribofuranosy|)imidazo|ecarboxamide, 5-amino(3-O-methyl-fi-D-ribofuranosy|)imidazolecarboxamide. 5-amino5-D-ribofuranosyiimidazoleN-[(4-nitrophenyl)methy1]car’ooxamide. 5-amino-1 -[5-D-ribofuranosylimidazoleN-[(5-chlorophenyl)methyflcarboxamide. 5-amino[3-D-ribofuranosylimidazole4-N-[(2,4dichloropheny1)methyl]carboxamide, 5-amino-1 -[5-chloro-Sdeoxy-fs-D-ribofuranosyl)imidazolecarboxamide. 5-amino[3-D-ribofuranosyiimidazoleN-[(3-nitropheny|)methyl]carboxamide. 5-aminc>1 -[3-D-ribofuranosylimidazoleN-[(4<:h|oropheny|)methyllcarboxamide, 5-amino(3-D-ribofuranosylimidazoleN-[(4-methylphenyl)methy|]carboxamide. 5-amino-1 -(3-O-ethyi-[3-D-ribofuranosylimidazole-carboxamide, 5-amino(2-O-n-butyl-[3-D-ribofuranosylimidazolecarboxamide. 5-amino-1 -(3-O-n-butyl-[5-D-ribofuranosylimidazolecarboxamide, 5-amino-1 -(2-O-ethyl-[3-D-ribofuranosylimidazole-carboxamide_ 5-aminoB-D-ribofuranosyiimidaioleN-[(3<:h|oropheny|)methyflcarboxamide. 5-amino(5-amino-Sdeoxyf5-D-ribofuranosyl)imidazole(N-cyclopentyl) carboxamide. 5-amino£5-D-ribofuranosyiimidazoleN-[4-methoxybenzyl]carboxamide. 5-amino5-D-ribofuranosylimidazole—4-N-(4-dimethylaminobenzyncarboxamide, 5-aminothiophenyl5-D-ribofuranosylimidazolecarboxamide. 5-amino[5-D-ribofuranosylimidazole—4-N-[(3-iodophenyI)methyflcarboxamide, 5-amino(3-D-ribofuranosyiimidazoleN-(2-endo-norborny|)carboxamide, 5-amino(5-iodo-5<1eoxy-fi-D-ribofuranosy1)imidazo|eN-[(4-nitrophenyl)methyl]carboxamide. 5-amino-1~fi-D-ribofuranosyiimidazoleN-[(1-(4-nitrophenyl)ethyl]carboxamide. 5-amino(5-chlorodeoxy-fi-D-ribofuranosynimidazoleN-[(4-nitropheny1)methyl]carboxamide. 5-amino-1 -(5-amino-Sdeoxy-1%D-ribofuranosyflimidazoleN-[(4-chlorophenyl)methyflcarboxamide. 5-amino(sdeoxymethylthio-[3-D-ribofuranosyI)imidazolecarboxamide. 5-aminoD-ribofuranosyiimidazoleN-(4-bromophenyI)carboxamide. 5-amino[3-D-ribofuranosylimidazoleN-[(4Joromopheny|)methyncarboxamide, 5-amino(5-D-ribofuranosylimidazoleN-(4-nitropheny1)carboxamide. 5-amino(5deoxy-f5-D-ribofuranosy|)imidazole-4~N-[(4-chlorophenyl)methy1]carboxamide, 5-amino(sdeoxymethylsutfiny|-f5-D-ribofuranosyI)imidazo|ecarboxamide. 5-amino5-D-(sdeoxymethylamino(5-D-ribofuranosyl)imidazo|e4—carboxamide. 5-amino-1 -fs-D-riboturanosyiimidazole4-N-(2-chlorophenyl)car‘ooxamide, 5-amino—1 -(5-deoxydiethylaminoribo-1%D-furanosyl)imidazo|ecarboxamide, 5-amino5-D-(5-benzylamino-5deoxy£5-D-ribofuranosyl)imidazole4-carboxamide. 5-aminothiop-D-(5-deoxyf3-D-ribofuranosyl)imidazo|ecarboxamide. 5-aminothio(5-aminodeoxy-[5-D-ribofuranosy|)imidazo|e-4<:ar’ooxamide, 5-amino(5-aminodeoxyD-ribofuranosyl)imidazoleN-[4-nitrophenyi)methyflcarboxamide, 5-amino5-D-ribofuranosyiimidazoleN-[3-(4-nitrophenyl)propyflcarboxamide, 5-amino5-D-ribofuranosyi)imidazoleN-[(4-trifluoromethylpheny|)methy|]carboxamide. 5-amino[5-D-ribofuranosyi)imidazoleN-[(4-suIfamoy1pheny|)methyI]carboxamide. 5-amino(5-(4-chlorobenzyl-aminodeoxyf5-D-ribofuranosyl)imidazo|ecarboxamide. 5-amino-(5-aminodeoxy-2.3-di-O-acetylD-ribofuranosyI)imidazoleN—[(4-chlorophenyl)methyI]carboxa- mide. and 5-amino(sdeoxysulfhydryl(5-D-ribofuranosyl)imidazolecarboxamide. 5-amino[3-D-ribofuranosylimidazoleN-[2-hydroxy(3.4—dihydroxyphenyl)ethy|]carboxamide. and 5-amino[3-D-ribofuranosylimidazoleN-[2-(4-nitrophenyl)ethyl]carboxamide. 130

50. A Compound according to claim 26 selected from 5—amino—4—(1—piperidinocarbamoy1)—1—fi-D- ribofuranosylimidazole, 5 5—aminofl-D—ribofuranosylimidazole-4—carboxy1ic acid p-nitrobenzylthio ester, and 5—amino[1-(4—nitrophenyl)]piperazino carbamoyl]-1—B—D—ribofuranosy1imidazole. 10A

51. A pharmaceutical composition comprising a compound according to any one of claims 26 to 50 and a pharmaceutically acceptable carrier, adjuvant or vehicle. F. R. KELLY & CO.,