Antiviral, antitumor, antimetastatic, immune system enhancing 7-thia-guanosine derivatives

Abstract: Use of a compound of the structure: wherein R₄, R₅, R₆ and R₇ individually are H, OH or C₁-C₁₈ O-acyl and R₃ is H, C₁-C₁₈ acyl or or R₅ and R₇ are H or OH, R₆ is H and together R₃ and R₄ are and X is =O or =S; Y is -OH, -SH, -NH₂ or halogen; and Z is H, -NH₂, -OH or halogen; wherein halogen is Cl or Br; or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for use in treating a virus in a mammal.

Description: TECHNICAL FIELD This invention is directed to the use of new and improved antiviral, nucleosides and nucleotides. BACKGROUND OF INVENTION The immune system is an inherently complex system that serves its host by providing natural resistance and recovery against both pathogens of an external source as well as aberrant 'self' cells, i.e. tumor growth. It provides both 'natural', i.e. inborn and unchanging, or 'acquired' , i.e. adaptive immune response. For the most part the immune system is innocuous to 'self.' The immune system is able, in most instances, to recognize 'self,' its host, and differentiate between 'self' and non-self. That is the immune system is 'self tolerant.' In certain instances, however, the immune system does attack its host as if it were foreign resulting in autoimmunity or autoimmune disease or hypersensitivity expressed in the form of allergies, certain forms of kidney disease and the like. While for the most part an effective and active immune system confers biological advantages for the host, modern medicine has sought in certain instances to repress the immune system because of autoimmunity hypersensitivity in graft or organ transplant and in other instances stimulate the immune system by immunization. It is therefore advantageous in certain instances to attempt to stimulate the immune system against pathogen or tumor attack and in other instances to repress the immune system when it becomes self destructive to the host or for organ transplant or the like. While most molecular entities either synthetic or natural which are known to stimulate the immune system are large molecules such as interferon, poly I:C or large messenger proteins, certain small molecules have also been shown to modulate the immune system as well. Of the small molecular entities the nucleoside 3-deaza-adenosine has been indicated in U. S. Patent 4,309,419 to Walberg, et al., which issued January 5, 1982, as being an inhibitor of the immune response. Other nucleosides, most notably 8-bromoguanosine, 8-mercaptoguanosine and 7-methyl-8-oxoguanosine have been noted as showing stimulation of the immune system. Certain components of the immune system are cellular in nature while others are humoral, that is they exist free in serum or other body fluids. Adaptive immunity is based upon special properties of lymphocytes. The lymphocyte populations are generally divided between T lymphocytes commonly called T cells and B lymphocytes commonly called B cells. The T lymphocytes undergo a maturation processing in the thymus whereas the B lymphocytes are continuously generated in the bone marrow and are responsible for the production of antibodies. The lymphocytes freely circulate in the blood and from blood gain access to the tissues from which they are collected and recycled back via the lymph systems including the lymph glands and spleen. Components of the cellular immune mechanisms include macrophages (hereinafter also referred to as MAC's), polymorphonuclear leukocytes commonly called PMN, mast cells and other cellular or molecular entities such as interferon and the like. Further, complements which are a series of proteins present in the serum can be activated by other immune components or directly by pathogens such as bacteria or the like. Natural killer cells, hereinafter also identified as NK cells, constitute a group of cells which are concerned with natural immunity. These are lymphoid cells which are generally found in at least the young of all mammalian species and can be readily elicited in older animals. They generally exert a selective cytotoxicity against a range of target cells mostly malignant tumor cells. The B cells produce antibody. Antibodies are a group of proteins of various classes including IgG, IgM, IgA, IgD, IgE. Not all specific antibody classes are present in different animal species. Generally the higher up on the evolutionary chain of animals the more antibody species present with warn blooded mammals generally having a full contingent of the different antibody species. The immune system is capable of modifying certain regions on the antibody proteins allowing the antibody protein to bind with specific antigens of various origins. These include pathogens, parts of pathogens such as cellular wall polysaccharide molecules, large protein or the like, as well as other foreign debris such as pollen and even in autoimmune diseases portions of the host itself. Some antibody production by the B cells is independent of the T cells while other antibody production is T cell dependent. There are several groups of T cells including helper T cells which stimulate other T cells and B cells for the production of antibody, suppressor T cells which modulate the immune response to keep it from overwhelming the host, cytotoxic T cells (CTL's) which are very important against pathogens especially viral pathogens and delayed hypersensitive T cells which are important in attracting and activating a variety of other cells, including the macrophages. The immune system is important in protecting the host against a variety of pathogens including bacteria, viruses, protozoa, parasitic worms such as flukes, tapeworms and round worms, fungi, and tumor cells of the host which become parasitic on the host. The antiviral activity of the immune system is generally associated with the T cells whereas the natural antitumor ability of the host resides with the macrophages, the natural killer cells, certain non-T and non-B myeloid cells and with certain portions of the complement system. As is evident, the immune system is a very complex system which is extremely important to the host for protection of the host against outside pathogens as well as against internal aberrant cells. Catastrophic effects to the host can result when pathogens, tumors or the like overwhelm the immune system of the host. It has even been suggested that tumors may have the ability to depress or subvert the hosts immune system. This is supported by the recognition of clinicians that viral and bacteria infections can be a major contributor to the deaths among tumor patients. In view of the above it is evident that there is a need for new and improved antiviral agents. BRIEF DESCRIPTION OF THE INVENTION The present invention relates to the use of a novel class of nucleosides and nucleotides of the thiazolo[4,5- d ] pyrimidine ring, system. In accordance with the invention, the use of compounds of the formula as follows is disclosed: wherein R 4 , R 5 , R 6 and R 7 individually are H, OH or C 1 -C 18 O-acyl and R 3 is H, C 1 -C 18 acyl or or R 5 and R 7 are H or OH, R 6 is H and together R 3 and R 4 are and X is =O or =S; Y is -OH, -SH, -NH 2 or halogen; and Z is H, -NH 2 , -OH or halogen; wherein halogen is Cl or Br; or a pharmaceutically acceptable salt thereof. A presently preferred group of compounds used in the invention are compounds of the formula: wherein R 1 and R 2 individually are H or C 1 -C 18 acyl and R 3 is H, C 1 -C 18 acyl or or R 1 is H and together R 2 and R 3 are and X is =O or =S; Y is -OH, -SH, -NH 2 or halogen; and Z is H, -NH 2 , -OH or halogen; wherein halogen is Cl or Br; or a pharmaceutically acceptable salt thereof. Particularly preferred are compounds wherein Z is -NH 2 , Y is -OH, X is =O or = S and R 1 , R 2 and R 3 each are H. The compounds used in the invention are useful as immune system enhancers and have certain immune system properties including modulation, mitogenicity, augmentation and/or potentiation or they are intermediates for compounds which have these properties. The compounds have been shown to express effects on at least the natural killer, and lymphocyte cells of the immune system of a host. Because of these properties they are useful as antiviral, agents or as intermediates for antiviral, agents. They can be used to treat an affected host by serving as the active ingredients of suitable pharmaceutical compositions. In accordance with the invention, compounds are utilized to treat viral diseases in host mammals by administering to the mammal a therapeutically effective amount of the compounds. Additionally in accordance with the invention, 5-amino-3-β- D -ribofuranosylthiazolo[4,5- d ]pyrimidine-2,7(6H)-dione is utilized to enhance natural killer immune cells in a host by administering to the host a therapeutically effective amount of 5-amino-3-β- D -ribofuranosylthiazolo[4,5- d ]pyrimidine-2,7(6H)-dione as the active component in a pharmaceutical composition. Additionally in accordance with the invention, 5-amino-3-β- D -ribofuranosylthiazolo[4,5- d ]pyrimidine-2,7(6H)-dione is utilized to enhance lymphocyte cells in a host by administering to the host a therapeutically effective amount of 5-amino-3-β- D -ribofuranosylthiazolo[4,5- d ] pyrimidine-2,7(6H)-dione as the active component in a pharmaceutical composition. Additionally in accordance with the invention a therapeutical pharmaceutical composition is disclosed which contains as its active ingredient a therapeutically effective amount of 5-amino-3-β- D -ribofuranosylthiazolo[4,5- d ]pyrimidine-2,7(6H)-dione. Additionally in accordance with the invention a prophylactic pharmaceutical composition is disclosed which contains as its active ingredient a prophylactically effective amount of 5-amino-3-β- D -ribofuranosylthiazolo[4,5- d ]pyrimidine-2,7(6H)-dione. In addition the prophylactic composition can include a further antiviral agent as a further active ingredient. In an advantageous process of the invention the compounds used in the invention are prepared by silylating a derivative of a 2,5,7-substituted thiazolo[4,5- d ]pyrimidine and reacting said silyl derivative with a 1- O -substituted-blocked- D -pentofuranose in the presence of a catalyst to form a blocked nucleoside. The blocking groups can further be removed from said nucleoside. Additionally the nucleoside can be phosphorylated. The substituent groups on said 2,5,7-substituted thiazolo[4,5- d ]pyrimidine are selected from the group consisting of H, halogen, -NH 2 , -OH, =O and =S. For certain compounds of the invention the substituted blocked pentofuranosyl sugar is selected as a 1- O -acetyl-2,3,5-tri- O -acyl- D -pentofuranose particularly 1- O -acetyl-2,3,5-tri- O -benzoyl- D -ribofuranose. DETAILED DESCRIPTION OF THE INVENTION The synthesis of the guanosine analog, 5-amino-3-β- D -ribofuranosylthiazolo[4,5-d]pyrimidine-2,7(6H)-dione, can be effected by the direct glycosylation of the preformed guanine base analog. (Scheme I). Thus, 5-aminothiazolo[4,5- d ]pyrimidine-2,7(3H,6H)-dione ( 4 ), prepared in five steps from the commercially available diaminopyrimidinone by the method of Baker and Chatfield, J. Chem. Soc. (C), 2478 (1970), was glycosylated by trimethylsilylation using hexamethyldisilazane followed by treatment with 1- 0 -acetyl-2,3,5-tri- 0 -benzoyl- D -ribofuranose ( 5 ) in the presence of trimethylsilyl trifluoromethanesulfonate as a catalyst. The major product, 5-amino-3-(2,3,5-tri- 0 -benzoyl-β- D -ribofuranosyl)thiazolo[4,5- d ]pyrimidine-2,7(6H)-dione ( 6 ) was isolated. Treatment of 6 with sodium methoxide in methanol gave the deprotected guanosine analogy 5-amino-3-β- D -ribofuranosylthiazolo[4,5- d ]pyrimidine-2,7(6H)-dione ( 7 ). When 7 was deaminated with excess nitrous acid the xanthosine analog, 3-β- D -ribofuranosylthiazolo[4,5- d ]pyrimidine-2,5,7(4H,6H)-trione (8) was produced. Replacement of the 5-amino group of compound 6 by a hydrogen atom was accomplished by treatment of 6 with t-butyl nitrite in tetrahydrofuran to yield 3-(2,3,5-tri- 0 -benzoyl-β- D -ribofuranosyl)thiazolo[4,5- d ]pyrimidine-2,7(6H)-dione ( 9 ). Deprotection of 9 using sodium methoxide in methanol or methanolic ammonia provided the inosine analog, 3-β- D -ribofuranosylthiazolo[4,5- d ]pyrimidine-2,7(6H)-dione ( 10 ). The guanosine analog, 7 , was phosphorylated and the 5'-monophosphate ( 11 ) was obtained. The 3',5'-cyclic monophosphate derivative, 12 , was then prepared from 11 . The preparation of the analogous 8-mercapto compound in the thiazolo[4,5- d ]pyrimidine system is depicted in Scheme II starting with 5-amino-2-chlorothiazolo[4,5- d ]-pyrimidin-7(6H)-one ( 13 ). Compound 13 was treated with NaSH in ethylene glycol at 110° to provide the 2-thioxo heterocycle, 14 . Glycosylation of 14 by the same procedure as that used to prepare the 2-oxo compound, 6 , (except that some heating was required to ensure that any S-glycoside formed would be converted to the more thermodynamically stable N-glycoside) resulted in the formation of 5-amino-2-thioxo-3-(2,3,5-tri- 0 -benzoyl-β- D -ribofuranosyl)thiazolo[4,5- d ]pyrimidin-7(6H)-one ( 15 ). Treatment of 15 with sodium methoxide in methanol yielded the 8-mercaptoguanosine analog, 5-amino-2-thioxo-3-β- D -ribofuranosylthiazolo[4,5- d ]pyrimidin-7(6H)-one ( 16 ). Various related derivatives in the guanosine analog series were also prepared. The 6-thioguanosine analog was prepared by two routes starting from 6 (Scheme III). In one approach, 6 was treated with the mild chlorinating agent dimethyl(chloromethylene)ammonium chloride (generated in situ from thionyl chloride and DMF), and provided 5-amino-7-chloro-3-(2,3,5-tri- 0 -benzoyl-β-D-ribofuranosyl)thiazolo[4,5- d ]pyrimidin-2-one( 17 ). Reaction of 17 with thiourea in refluxing ethanol gave the protected thioguanosine analog, 5-amino-7(6H)-thioxo-3-(2,3,5-tri- 0 -benzoyl-β- D -ribofuranosyl)thiazolo[4,5- d ]pyrimidin-2-one (18). Compound 18 was also prepared directly from 6 by reaction with P 2 S 5 in pyridine. Deprotection of 18 was accomplished either with sodium methoxide in methanol or with methanolic ammonia and the 6-thioguanosine analog, 5-amino-7(6H)-thioxo-3-β-D-ribofuranosylthiazolo[4,5,- d ]pyrimidin-2-one ( 19 ) was isolated as the crystalline monohydrate. The chloro function at position 7 was also nucleophilically substituted by azide using sodium azide in dry DMF which subsequently ring closed onto N-6 to form the new tricyclic ring compound, 5-amino-7-(2,3,5-tri- 0 -benzoyl-β- D -ribofuranosyl)tetrazolo[1,5- c ]thiazolo[4,5- d ]pyrimidin-2-one ( 20 ). In an effort to study the thiazolo[4,5- d ]pyrimidine ring system with respect to the order of nucleophilic substitution at the 2,5, and 7 positions and possibly use this information to synthesize the adenosine analog, chlorination of the readily available 2-chlorothiazolo[4,5- d ]pyrimidine-5,7(4H,6H)-dione ( 21 ) using refluxing POCl 3 and N,N-dimethylaniline (Scheme IV) was effected. The desired 2,5,7-trichlorothiazolo[4,5,- d ]pyrimidine ( 22 ) was obtained along with a small amount of 5,7-dichloro-2-(N-methylanilino)thiazolo[4,5- d ]pyrimidine ( 23 ). The trichloro compound, 22 , was carefully hydrolyzed in 1N NaOH at 60°C in order to obtain the mono-oxo derivative, 5,7-dichlorothiazolo[4,5- d ]pyrimidin-2(3H)-one ( 24 ), the structure of which was verified by single crystal X-ray analysis. Reaction of 24 with 1,2,3,5-tetra- 0 -acetyl- D -ribofuranose ( 25 ) under fusion glycosylation conditions produced 5,7-dichloro-3-(2,3,5-tri- 0 -acetyl-β- D -ribofuranosyl)thiazolo[4,5- d ]pyrimidin-2-one ( 26 ). Attempts to use 26 for further modification to obtain the adenosine analog were unsuccessful due to the labile nature of the thiazole ring toward nucleophilic ring-opening. This was circumvented by the synthesis of the adenosine analog from its preformed heterocycle in the same manner as that used to obtain the guanosine analog. The known 2,7-diaminothiazolo[4,5- d ]pyrimidine ( 27 ) served as the starting material (Scheme V). Treatment of 27 with nitrous acid under conditions similar to those used to prepare 13 provided 7-amino-2-chlorothiazolo[4,5- d ]pyrimidine ( 28 ). The structure of compound 28 was verified by single-crystal X-ray analysis. Treatment of 28 with NaSH in DMF at 0°C yielded the 2-mercapto derivative, 7-aminothiazolo[4,5- d ]pyrimidine-2(3H)-thione ( 29 ). The conversion of the 2-thioxo function in 29 to a 2-oxo function was accomplished using cold alkaline hydrogen peroxide to yield 7-aminothiazolo[4,5- d ]pyrimidin-2(3H)-one ( 30 ). Reaction of 30 with the benzoyl-protected sugar, 5 , under the same glycosylation conditions (at room temperature) as used to produce the blocked guanosine analog, 6 , resulted in the formation of the unexpected blocked 4-ribofuranosyl isomer, 7-amino-4-(2,3,5-tri- 0 -benzoyl-β- D -ribofuranosyl)thiazolo[4,5- d ]pyrimidin-2(3H)-one ( 31 ), as the only isomer detected and isolated. If, however, the same reaction was carried out at elevated temperature (80°C), the predominant product obtained was the desired 3-ribofuranosyl isomer, 7-amino-3-(2,3,5-tri- 0 -benzoyl-β- D -ribofuranosyl)thiazolo[4,5- d ]pyrimidin-2(3H)-one ( 32 ). Both isomers, 31 and 32 , were deprotected using sodium methoxide in dry methanol to obtain 7-amino-4-β- D -ribofuranosylthiazolo[4,5- d ]pyrimidin-2(3H)-one ( 33 ) and 7-amino-3-β- D -ribofuranosylthiazolo[4,5- d ]pyrimidin-2(3H)-one ( 34 ), respectively. The 2'-deoxyerythropentofuranosyl, the xylofuranosyl and the arabinofuranosyl analogs of compound 7 can be prepared in a manner similar to that described for compound 7 (scheme VI). Thus reacting 4 with 1-chloro-2-deoxy-3,5-di- O -(p-toluoyl)-α- D -erythropentofuranose followed by deblocking yields 5-amino-3-(2-deoxy-β- D -erythropentofuranosyl)thiazolo[4,5- d ]pyrimidine-2,7(3H,6H)-dione ( 36 ). Likewise after deblocking of the respective intermediates 4 and 1,2,3,5-tetra- O -acetyl- D -xylofuranose will yield 5-amino-3-(β- D -xylofuranosyl)thiazolo[4,5- d ]pyrimidine-2,7(3H,6H)-dione ( 38 ) and 4 and 1-chloro-2,3,5-tri- O -benzyl-α -D -arabinofuranose will yield 5-amino-3-(β- D -arabinofuranosyl)thiazolo[4,5- d ]pyrimidine-2,7(3H,6H)-dione ( 40 ). The tri- O -acetyl ( 40 ) prodrug form of compound 7 is prepared from 4 and 1,2,3,5-tetra- O -acetyl- D -ribofuranose and the 2,3-di- O -isopropylidene derivative ( 42 ) is prepared directly from 7 (scheme VI). For the compounds used in the invention, pharmaceutically acceptable acid addition salts of the basic moiety can be selected from, but not necessarily limited to, the group consisting of hydrochloride, hydrobromide, hydroiodide, citrate, sulfate, substituted sulfate, phosphate, carbonate, bicarbonate and formate. Pharmaceutically acceptable salts of the phosphate moiety can be selected from, but not necessarily limited to, the group consisting of alkali and alkaline earths, e.g. sodium, potassium, calcium, magnesium, lithium, ammonium and substituted ammonium, trialkylammonium, dialkylammonium, alkylammonium, e.g. triethylammonium, trimethylammonium, diethylammonium, octylammonium, cetyltrimethylammonium and cetylpyridium. The hydroxyl groups on the glycon and heterocycle and the amino groups of the heterocycle can be blocked with groups such as, but not necessarily limited to, acyl, isopropylidene and dimethylaminomethylene. The acyl group can be selected from a group consisting of C 1 -C 18 straight chain, branched ,chain, substituted, unsaturated, saturated or aromatic acid such as, but not necessarily limited to acetic, trifluoroacetic, propionic, n-butyric, isobutyric, valeric, caproic, pelargonic, enanthic, caprylic, lactic, acrylic, propargylic, palmitic, benzoic, phthalic, salicylic, cinnamic and naphthoic acids. Melting points were taken on a Thomas-Hoover capillary melting point apparatus or on a Haake-Buchler digital melting point apparatus and are uncorrected. Nuclear magnetic resonance ( 1 H NMR) spectra were determined at 300.1 MHz with an IBM NR300AF spectrometer. The chemical shifts are expressed in δ values (parts per million) relative to tetramethylsilane as internal standard. Ultra violet spectra (UV: sh = shoulder) were recorded on a Beckman DU-50 spectrophotometer. Elemental analyses were performed by Robertson Laboratory, Madison, N.J. Evaporations were carried out under reduced pressure with the bath temperature below 40°C. Thin layer chromatography (TLC) was run on silica gel 60 F-254 plates (EM Reagents). E. Merck silica gel (230-400 mesh) was used for flash column chromatography. EXAMPLE 1 5-amino-3-(2,3,5-tri-O-benzoyl-β-D-ribofuranosyl)thiazolo[4,5-d]pyrimidine-2,7(6H)-dione ( 6 ). A mixture of dry 5-aminothiazolo[4,5-d]pyrimidine-2,7-(3H,6H)-dione 4 (5.5 g, 30 mmol), hexamethyldisilazane (HMDS, 100 mL), ammonium sulfate (15 mg) and pyridine (10 mL) was heated under reflux for 4 h with the exclusion of moisture. Excess HMDS was removed by distillation to provide the syrupy bis-silyl derivative. The bis-silyl intermediate was dissolved in dry acetonitrile (300 mL) and 1- 0 -[acetyl-2,3,5-tri- 0 -benzoyl- D -ribofuranose ( 5 : 15.1 g, 30 mmol) was added followed by trimethlsilyl trifluoromethane sufonate (9.3 mL, 42 mmol). The clear reaction mixture was stirred at ambient temperature for 16 h. The solvent was evaporated to dryness and the residual syrup was dissolved in EtOAc (600 mL). The solution was washed with 5% NaHCO 3 solution (2 x 150 mL), and the dried (Na 2 SO 4 ) organic layer was evaporated. The residual syrup was triturated with ether to yield 18.1 g (96%). The resulting foam was purified on a silica gel column by Prep LC techniques using CHCl 3 -MeOH (9:1, v/v) as the solvent. Recrystallization of the residue from EtOH gave 5-amino-3-(2,3,5-tri- 0 -benzoyl-β- D -ribofuranosyl)thiazolo[4,5- d ]pyrimidine-2,7(6H)-dione ( 6 ) as colorless cyrstals: yield 14.5 g, 77%: mp 248-250°C: UV λ max (pH 1) 215 sh nm (ε 28000), 219 (28000), 224 sh (27600), 301 (8500): UV λ max (pH 7) 215 sh nm (ε 28900), 222 (29500), 301 (10600): UV λ max (pH 11) 218 nm (ε 27800), 273 (6900): Anal. Calcd. for C 31 H 2 N 4 O 9 S: C, 59.23: H, 3.85: N, 8.91: S, 5.10. Found: C, 59.26: H, 3.89: N, 8.93: S, 5.23. EXAMPLE 2 5-Amino-3-β- D -ribofuranosylthiazolo[4,5-d]pyrimidine-2,7(6H)-dione ( 7 ). A solution of 6 (0.75 g, 1 mmol) in methanol (75 mL) was adjusted to pH 9 with NaOCH 3 and stirred at room temperature for 16 h. The reaction mixture was evaporated to dryness and the residue was triturated with ether (2 x 75 mL). The ether insoluble solid was dissolved in water (15 mL) and acidified with acetic acid whereupon the crude product precipitated. Crystallization of this material from water gave a colorless powder: yield 0.31 g, 78%: mp 238°C (decomp.): UV λ max (pH 1) 215 nm (ε 2280), 245 (6900), 301 (8400): UV λ max (pH 7) 215 nm (ε 22100), 245 (6900), 301 (8000): UV λ max (pH 11) 245 nm (ε 5700), 291 (6000). NMR (DMSO-d 6 ) δ 5.79 (1H, d, J = 5.32 Hz, C 1 , H), 6.90 (2H, s, NH 2 ), 11.12 (1H, s, NH), and other sugar protons. Anal. Calcd. for C 10 H 12 N 4 O 6 S.H 2 O: C, 35.92: H, 4.22: N, 16.76: S, 9.59. Found: C, 35.82: H, 4.02: N, 16.92: S, 9.66. EXAMPLE 3 3-β-D-Ribofuranosylthiazolo[4,5-d]pyrimidine-2,5,7(4H,6H)-trione ( 8 ). To a suspension of 7 (0.76 g, 2.4 mmol) in glacial acetic acid (150 mL) was added dropwise a solution of sodium nitrite (1.5 g, 21.7 mmol) in water (15 mL) with stirring. After 30 min the suspension became clear and stirring was continued at room temperature overnight. The white solid which had separated was filtered, washed with cold water and dried. Recrystallization from hot water gave fine colorless crystals of 8 : yield 0.3 g, 40%: mp 250°C dec.: UV λ max (pH 1) 293 nm (ε 5500): UV λ max (pH 7) 212 nm (ε 14200), 301 (6100): UV λ max (pH 11) 204 nm (ε 21900), 301 (5600). Anal. Calcd. for C 10 H 11 N 3 O 7 S: C, 37.86: H, 3.49: N, 13.24: S, 10.10. Found: C, 37.81: H, 3.42: N, 13.01: S, 10.01. EXAMPLE 4 3-(2,3,5-Tri-O-benzoyl-β-D-Ribofuranosyl)thiazolo[4,5-d]pyrimidine-2,7(6H)-dione ( 9 ). To a solution of 6 (6.65 g, 10.6 mmol) in dry THF (350 mL) was added ter -butyl nitrite (6.2 mL, 52.3 mmol) and the mixture was stirred at room temperature for 1 h. Additional nitrite reagent (2.0 mL) was added and the mixture was stirred at 50-60°C overnight. The mixture was evaporated and the residue was purified by flash column chromatography on silica gel using 8-10% acetone in CH 2 Cl 2 followed by 10-11%. The desired product eluted last to yield 3.45 g (46%) of 9 as a foam: UV λ max (EtOH) 220 nm (ε 46600), 259 sh (11000), 271 sh (8400): 1 H NMR (DMSO-d 6 ) δ 6.31 (d, J = 6.45 Hz, 1H, C 1 , H), 7.38-7.98 (m, 15H, benzoyl aromatics), 8.25 (s, 1H, C 5 H), 13.16 (b, 1H, N 6 H, exchanged with D 2 O), and other sugar protons. EXAMPLE 5 3-β-D-Ribofuranosylthiazolo[4,5-d]pyrimidine-2,7(6H)-dione ( 10 ). Compound 9 (1.0 g, 1.63 mmol) was combined with methanolic ammonia (saturated at O°C, 50 mL) and heated at 90°C for 14 h in a steel bomb. The solvent was evaporated and the residue was treated with hot benzene which was decanted off. The resulting solid was purified by silica gel flash chromatography using chloroform and then CHCl 3 -MeOH (6:1) to yield 280 mg (57%) of 10 after crystallization from water: mp 216-218°C: UV λ max (pH 1) 217 nm (ε 25300), 259 (9700), 286 (6300): 1 H NMR (DMSO-d 6 ) δ 5.85 (d, J = 5.1 Hz, 1H, C 1 , H), 8.30 (s, 1H, C 5 H), 13.09 (b, 1H, N 6 H, exchanges with D 2 O), and other sugar protons. Anal. Calcd. for C 10 H 11 N 3 O 6 S: C, 39.87: H, 3.68: N, 13.95: S, 10.64. 3.61: N, 14.06: S, 10.43. EXAMPLE 6 5-Amino-3-β-D-Ribofuranosylthiazolo[4,5-d]pyrimidine-2,7(6H)-dione 5'-Monophosphate Ammonium Salt ( 11 ). To a suspension of 7 (2.1 g, 6.6 mmol) in freshly distilled trimethyl phosphate (25 mL) at -20°C was added POCl 3 (0.64 mL, 6.6 mmol) and then an additional equivalent of POCl 3 after 1 h. The mixture was stirred at -5°C for another 2 h and then poured into ethyl ether (150 mL, anhydrous) and centrifuged (6000 rpm, 10 min). The ether layer was decanted and ice water (100 mL) was added to the residual oil. The pH of the resulting solution was adjusted to 7.5 with aqueous ammonium bicarbonate and the solution was applied to a DEAE-cellulose column (3.2 x 35 cm), washed with water and eluted with a gradient (0 to 0.25 M, 2 L reservoirs) of aqueous ammonium bicarbonate. During the water wash, unreacted starting material eluted off (0.5 g). The appropriate fractions were pooled, evaporated and lyophilized several times to yield 1.01 g (48%, based on reacted starting material): mp 190-194°C UV λ max (pH 1,7) 243 nm (ε ), 301 (): UV λ max (pH 11) 243 nm (ε ), 289 () : 1 H NMR (DMSO-d 6 ) δ 5.71 (s, 1H, C 1 ,H), 7.15 (b, 5H, NH 2 and NH 4 + , exchanges with D 2 O), 11.25 (b, 1H, N 6 H, exchanges with D 2 O), and other sugar protons. Anal. C 10 H 14 N 5 O 8 SP.1.25H 2 O: C, 28.75: H, 3.98: N, 16.76: S, 7.67: P, 7.41. Found: C, 29.15: H, 3.68: N, 16.39: S, 7.76: P, 7.22. EXAMPLE 7 5-Amino-3-β-D-Ribofuranosylthiazolo[4,5-d]pyrimidine-2,7(6H)-dione 3',5'-Cyclic Monophosphate Ammonium Salt ( 12 ). Compound 11 (1.02 g, 2.28 mmol) was dissolved in water (10 mL) and pyridine (3 mL) and morpholino-dicyclohexylcarbodiimide (667 mg, 2.28 mmol) was added. The solution was evaporated and co-evaporated to a syrup several times with dry pyridine. After drying overnight over P 2 O 5 under vacuum, the syrup was dissolved in dry pyridine (100 mL) and added dropwise to a refluxing solution of pyridine (300 mL) containing DCC (25 g). The solution was refluxed for an additional 2 h, cooled and allowed to stir overnight. The mixture was evaporated to dryness and the residue was partitioned between water (150 mL) and ethyl ether (150 mL). The aqueous layer was concentrated to about 100 mL and applied, having pH 7.7, to a DEAE-cellulose column (3.2 x 30 cm) and washed with water followed by elution using a gradient of aqueous ammonium bicarbonate (0 to 0.19 M). The proper fractions were collected based on UV monitoring, evaporated and lyophilized several times to yield 190 mg (22%) of the title compound: mp 244°C (dec.): UV λ max (pH 1,7) 243 nm (ε ), 300 (): UV λ max (pH 11) 243 nm (ε ), 289 () : 1 H NMR (DMSO-d 6 ) δ 5.60 (d, J = 4.44, 1H, 2'OH, exchanges with D 2 O), 5.72 (s, 1H, C 1 , H), 7.15 (b, 6H, NH 2 and NH 4 , exchanges with D 2 O), 11.40 (b, 1H, N 6 H, exchanges with D 2 O), and other sugar protons. Anal. Calcd. for C 10 H 11 N 4 O 8 SP.NH 3 .1.25H 2 O: C, 28.75: H, 3.98: N, 16. Found: C, 29.15: H, 3.68: N, 16.39: S, 7.76: P, 7.22. EXAMPLE 8 5-Amino-2-thioxothiazolo[4,5-d]pyrimidin-7(6H)-one ( 14 ). A suspension of 5-amino-2-chlorothiazolo[4,5- d ]pyrimidin-7(6H)-one ( 13 : 1.5 g, 7.4 mmol) in ethylene glycol (30 mL) was heated to 110°C and NaSH x H 2 O (420 mg, 74 mmol) was added. A clear solution was not obtained, however, until an additional 250 mg were added. The clear solution was stirred at 110°C for 2 h and then the reaction mixture was cooled to room temperature, poured into ice (300 mL), and the pH adjusted to 2-3 with 10% HCl. The resulting pink gelatinous mixture was boiled for 1 h and the pink solid was collected by filtration through a medium frit-glass filter, washed with water and dried: yield 1.2 g, 81%: an analytical sample was prepared by flash column chromatography using EtOAc-MeOH-H 2 O-acetone (7:1:1:1). mp >300°C: UV λ max (pH 1) 243 nm (ε 13700), 266 (16500), 351 (17200): UV λ max (pH7) 262 nm (ε 14800), 345 (12700): UV λ max (pH 11) 250 nm (ε 19300), 335 (14300): 1 H NMR (DMSO-d 6 ) δ 6.91 (s, 2H, NH 2 ), 11.18 (s, 1H, N 6 H), 13.78 (s, 1H, N 3 H). Anal. Calcd. for C 5 H 4 N 4 OS 2 : EXAMPLE 9 5-Amino-2-thioxo-3-(2,3,5-tri-O-benzoyl-β-D-ribofuranosyl)thiazolo[4,5-d]pyrimidin-7(6H)-one ( 15 ). Compound 14 (1.0 g, 5 mmol) was glycosylated in the same manner as that used to prepare 6 , requiring HMDS (20 mL), benzoyl-blocked sugar ( 5 : 2.52 g, 5 mmol), and TMS-triflate (1.45 mL, 7.5 mmol). At the end of the 16 h reaction period, the reaction mixture was heated at 70°C for 3 h in order to rearrange any S-glycoside formed to the more stable N-glycoside. After the same workup, 15 (2.1 g crude) was purified by flash column chromatography using hexanesacetone (1:1) and crystallized from toluene-EtOAc: yield 1.9 g, 59%: mp 230-233°C (darkens 195°C). Anal. Calcd. for C 31 H 24 N 4 O 8 S 2 : C, 57.76: H, 3.75: N, 8.69: S, 9.95. Found: C, 57.98: H, 3.46: N, 8.40: S, 9.66. EXAMPLE 10 5-Amino-2-thioxo-3-β-D-ribofuranosylthiazolo[4,5-d]pyrimidin-7(6H)-one ( 16 ). To a solution of 15 (1.25 g, 1.94 mmol) in dry methanol (100 mL) was added sodium methoxide powder until the pH reached 10. The solution was stirred overnight and then neutralized with Dowex H + resin and filtered. After evaporation of the filtrate, the residue was washed with ether to remove methyl benzoate and the crude material was crystallized from water: yield 520 mg, 81%: mp 220°C dec.: UV λ max 1 H NMR (DMSO-d 6 ) δ 6.48 (d, J = 3.00 Hz, 1H, C 1 ,H), 6.99 (s, 2H, NH 2 ), 11.47 (s, 1H, NH), and other sugar protons. Anal. Calcd. for C 10 H 12 N 4 O 5 S 2 .H 2 O: C, z34.28: H, 4.03: N, 15.99: S, 18.30. Found: C, 33.99: H, 3.92: N, 15.68: S, 18.22. EXAMPLE 11 5-Amino-7-chloro-3-(2,3,5-tri-O-benzoyl-β-D-ribofuranosyl)thiazolo[4,5-d]pyrimidin-2-one ( 17 ). Dry purified 6 (10 g, 16 mmol) was dissolved in dry methylene chloride (350 mL) and a solution of freshly distilled thionyl chloride (40 mL), dry DMF (20 mL), in dry methylene chloride was added dropwise over a 2 h period and the reaction was kept at 60°C (reflux) for 16 h. The reaction mixture was poured carefully into ice and NaHCO 3 solution and stirred for 30 min. The layers were separated and the aqueous layer extracted (2 x 150 mL) with methylene chloride and the combined layers dried over Na 2 SO 4 and evaporated in vacuo . The residual syrup was purified by passing through a silica gel column (4 x 40 cm) and eluting with CHCl 3 -acetone (4:1), to obtain the chloro compound as a white foam, 8.6 g, 84%: mp 88-90°C: Anal. Clcd. for C 31 H 23 C 1 N 4 O 8 S: C, 57.54: H, 3.58: Cl, 5.47: N, 8.66: S, 4.96. Found: C, 58.06: H, 3.99: Cl, 5.95: N, 9.41: S, 4.75. EXAMPLE 12 5-Amino-7(6H)-thioxo-3-(2,3,5-tri-O-benzoyl-β-D-ribofuranosyl)thiazolo[4,5-d]pyrimidin-2-one ( 18 ). Method 1 A mixture of 17 (3.3 g, 5 mmol), thiourea (0.719 g, 1 mmol) and EtOH (100 mL) was heated under reflux for 6 days. The reaction mixture was evaporated, and the residue was extracted with CHCl 3 (200 mL). The solvent was evaporated to dryness in vacuo , and the residue was purified by silica gel column chromatography with CHCl 3 -acetone (7:1) as the eluant. After evaporation the residue was crystallized from EtOH to afford a colorless powder: yield 1.9 g, 58%: mp 227-229°C: UV λ max (pH 1) 234 nm (ε 26000), 280 sh (9000), 365 (11800): UV λ max (pH 11) 230 nm (ε 40500), 267 (8700), 327 (14100): Anal. calcd. for C 31 H 24 N 4 O 8 S 2 : C, 57.75: H, 3.75: N, 8.69: S, 9.95. Found: C, 57.79: H, 3.79: N, 8.69: S, 9.98. Method 2 To a solution of 6 (1 g, 1.6 mmol) in pyridine (50 mL) was added with stirring P 2 S 5 (1.5 g, 6.2 mmol). The solution was refluxed gently (bath temperature 130-140) for 29 h. The reaction mixture was evaporated to dryness in vacuo. The excess P 2 S 5 was decomposed by the addition of H 2 0 (200 mL) at 60°C. The mixture was stirred for 1 h, then left at room temperature overnight. The resulting solid was filtered, dissolved in CHCl 3 , dried (Na 2 SO 4 ) and the solvent removed under vacuum. The residue was purified by silica gel column chromatography with CHCl 3 -acetone 7:1 as the eluant. After concentration the residue was crystallized from EtOH to give 18 (0.43 g, 43%). The physicochemical properties of compound 18 prepared by Method 2 were found to be identical in all respects to those of the compound prepared by Method 1 above. EXAMPLE 13 5-Amino-7(6H)-thioxo-3-β-D-ribofuranoxylthiazolo[4,5-d]pyrimidin-2-one ( 19 ). Method 1 A solution of 18 (1 g, 1.6 mmol) in methanol (50 mL) was adjusted to pH 9 with NaOCH 3 and stirred at room temperature for 16 h. The reaction mixture was evaporated to dryness and the residue was triturated with ether (2 x 75 mL). The ether insoluble solid was dissolved in water (15 mL) and acidified with acetic acid whereupon the crude product was precipitated. Recrystallization of this material from EtOH-H 2 O gave colorless prisms: yield 0.47 g, 87%: mp 185-187°C: UV λ max (pH 1) 214 nm (ε 2700), 230 sh (14000), 263 (6700), 354 () : UV λ max (pH 7) 213 nm (ε 25900), 247 (9100), 266 sh (7700), 334 (12100), 353 (11800): UV λ max (pH 11) 247 nm (ε 12300), 266 sh (8800), 327 (16100): 1 H NMR (DMSO-d 6 ) δ 5.76 (d, J = 5.32 Hz, C 1 ,H) 7.22 (s, 2H, NH 2 ), 12.41 (s, 1H, NH), and other sugar protons. Anal. Calcd. for C 10 H 12 N 4 O 5 S 2 .H20 : C, 34.28: H, 4.03: N, 15.99: S, 18.30. Found: C, 34.28: H, 3.99: N, 16.24: S, 18.51. Method 2 A solution of 18 (1.0 g, 1.6 mmol) in methanolic ammonia (saturated at O°C, 60 mL) was stirred at room temperature for 48 h. The solvent was evaporated to dryness and the residue was triturated with boiling benzene (2 x 100 mL) . The benzene insoluble solid was crystallized from EtOH-H 2 O to give 19 (0.36 g, 67%). The compound prepared by this method was identical to compound 19 prepared by Method 1 above, as judged by spectral and physical data. EXAMPLE 14 5-Amino-7-(2,3,5-tri-O-benzoyl-β-D-ribofuranosyl)tetrazolo[1,5-c]thiazolo[4,5-d]pyrimidin-2-one ( 20 ). To a solution of 17 (3.0 g, 4.6 mmol) in dry DMF (30 mL) was added sodium azide (0.3 g, 4.6 mmol) and the mixture was stirred at room temperature for 3 days. After evaporation of the solvent, the residue was dissolved in EtOAc (250 mL) and washed with water (2 x 50 mL), dried over sodium sulfate and evaporated. The resulting foam was purified by silica gel column chromatography using CHCl 3 -acetone 7:1. The product was crystallized from EtOH to give a white powder: yield, 2.0 g, 67%: mp 112-114°C: IR showed no azide band in the region of 2100 to 2200 cm -1 : UV λ max (MeOH). Anal. Calcd. for C 31 H 23 N 7 O 8 S: C, 56.96: H, 3.55: N, 15.00: S, 4.91. Found: C, 57.19: H, 3.88: N, 15.26: S, 4.75. EXAMPLE 15 2,5,7-Trichlorothiazolo[4,5-d]pyrimidine ( 22 ). A mixture of 2-chlorothiazolo[4,5-d]pyrimidine-5,7(4H,6H)-dione ( 21 , 15.8 g, 78 mmol), POCl 3 (220 ml) and N,N-dimethylaniline (12.3 g, 0.1 mmol) was refluxed for 3 h. The excess POCl 3 was removed under reduced pressure and the residue was poured into ice-water (500 mL) with stirring. The resulting aqueous solution was extracted with CHCl 3 (3 x 400 mL) and the organic layer was washed with water (2 x 400 mL), 0.1 N NaOH (2 x 300 mL) and water (2 x 400 mL) successively and then dried over Na 2 SO 4 . Evaporation of the chloroform produced a residue which was purified by silica gel column chromatography using CHCl 3 to provide the title compound ( 22 ) after crystallization from EtOH. Yield 13.8g, 74%: mp. 121-122°C: UV λ max (pH 1,7,11) 296 nm (ε 10,800). Anal. Calcd. for C 5 Cl 3 N 3 S: C, 24.97: Cl, 44.22: N, 17.48. Found: C, 25.02: Cl, 44.39: N, 17.37. EXAMPLE 16 5,7-Dichlorothiazolo[4,5-d]pyrimidin-2(3H)-one ( 24 ). A suspension of the trichloro compound (22: 3.0 g, 12 mmol) in 1N NaOH (35 mL) was heated at 60°C for 1 h. The solution was treated with decolorizing carbon and then acidified with 10% aqueous HCl. The resulting precipitate was collected and reprecipitated from dilute base with glacial acetic acid to provide 24 as orange needles (1.38g, 50%): mp. 191-192°C: UV λ max (pH 1) 254 nm (ε 5,300), 290 (11,400): UV λ max (pH 7,11) 226 nm (ε 28,900), 300 (14,100). Anal. Calcd. for C 5 HCl 2 N 3 OS: C, 27.04: H, 0.45: Cl, 31.93: N, 18.93. Found: C, 26.78: H, 0.61: Cl, 32.15: N, 18.66. Single crystal X-ray analysis of 24 showed the structure assignment to be correct. EXAMPLE 17 5,7-Dichloro-3-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)thiazolo[4,5-d]pyrimidin-2-one ( 26 ). A finely powdered mixture of 24 (3.7 g, 16 mmol), 1,2,3,5-tetra-O-acetyl- D -ribofuranoase (5.3 g, 16 mmol) and bis-( p -nitrophenyl)phosphate (20 mg) was heated at 170°C for 10 min under reduced pressure. After cooling to room temperature the brown solid mass was dissolved in EtOAc (500 mL) and washed with saturated aqueous sodium bicarbonate (3 x 300 mL). The dried (Na 2 SO 4 ) organic layer was evaporated to yield a syrup which was purified by silica gel column chromatography (4 x 40 cm) using toluene-EtOAc (5:1). The resulting syrup was crystallized from ethanol to give a colorless powder: yield 6.4 g, 80%: mp 125-126°C: 1 H NMR (DHSO-d 6 ) δ 1.99, 2.06, 2.08 (3s, 9H, acetyl), 6.07 (d, J = 3.40 Hz, 1H, C 1 ,H), and other sugar protons. Anal. Calcd. for C 16 H 15 Cl 2 N 3 0 8 S: C, 40.01: H, 3.15: Cl, 14.76: N, 8.75: S, 6.68. Found: C, 40.20: H, 3.31: Cl, 14.79: N, 8.61: S, 6.66. EXAMPLE 18 7-Amino-2-chlorothiazolo[4,5-d]pyrimidine ( 28 ). To a suspension of 2,7-diaminothiazolo[4,5-d]pyrimidine ( 27 : 16.3 g, 97.3 mmol) in water (200 mL) at 55°C was added enough 1 N NaOH (about 100 mL) to dissolve the starting material and sodium nitrite (8.0 g) was then added. This solution was then added dropwise over 30 min. to a solution containing con HCl (400 mL), water (100 mL) and LiCl (60 g) at 30°C. The resulting mixture was warmed to 45°C for 15 min. and then hot water (1 L, 90°) was added. The reaction mixture was stirred overnight at room temperature, filtered to remove unreacted starting material and the filtrate was neutralized with solid NaOH to pH 4. The resulting solid was filtered off, washed with water and dried to yield 28 : 5.38 g, 34%: recrystallization from water gave an analytical sample: mp >234°C decomp.: UV λ max (pH 1) 228 nm (ε ), 296 (): UV λ max (pH 7) 232 nm (ε ), 298 (): UV λ max (pH 11) 227 nm (ε ), 300 () : 1 H NMR (DMSO-d 6 ) δ 7.82 (b, 2H, NH 2 , exchanges with D 2 O), 8.41 (s, 1H, C 5 H). Anal. Calcd. for C 5 H 3 N 4 SCl.0.1H 2 O: C, 31.87: H, 1.71: N, 29.74: S, 17.02: Cl, 18.82. Found: C, 31.71: H, 1.50: N, 29.35: S, 16.92: Cl, 19.54. EXAMPLE 19 7-Aminothiazolo[4,5-d]pyrimidine-2(3H)-thione ( 29 ). A suspension of compound 28 (1.11 g, 5.9 mmol) in dry DMF (10 mL) was cooled in an ice bath to O°C and NaSH x H 2 O (0.87 g, 11.8 mmol) was added. The resulting clear solution was stirred overnight at 0°C and then at room temperature for 2h. The reaction mixture was poured into ice (300 mL) and the pH adjusted to 3-4 with glacial acetic acid. The solid precipitate was filtered, washed with water and dried to yield 0.96 g (88%). An analytical sample was prepared by crystallization from DMF-water: mp >370°C: UV λ max (pH 1) 248 nm (ε ), 263 (), 345 (): UV λ max (pH 7, 11) 228 nm (ε ), 258 (), 329 (): 1 NMR (DMSO-d 6 ) δ 7.57 (b, 1H, NH 2 , exchanges with D 2 O), 8.23 (s, 1H, C 5 H), 14.13 (b, 1H, N 3 H, exchanges with D 2 O). Anal. Calcd. for C 5 H 4 N 4 S 2 : C, 32.60: H, 2.19: N, 30.41: S, 34.81. Found: C, 32.97: H, 2.13: N, 30.29: S, 34.59. EXAMPLE 20 7-Aminothiazolo[4,5-d]pyrimidin-2(3H)-one ( 30 ). To a suspension of 29 (770 mg, 4.2 mmol) in water (30 mL) was added 1N NaOH (4.2 mL) and 30% H 2 O 2 (1.0 mL) and the reaction was stirred for 1h at room temperature. Additional peroxide (2.0 mL) and hydroxide (5.0 mL) were added and the mixture was stirred for 1h at 70°C. The reaction mixture was filtered and the filtrate was neutralized with glacial acetic acid. The resulting precipitate was filtered off while still hot, washed with cold water and dried to yield 0.52 g (74%): mp >370°C: UV λ max (pH 1) 267 nm (ε ), 289 (): UV λ max (pH 7, 11) 285 nm (ε ): 1 H NMR (DMSO-d 6 ) δ 7.18 (b, 2H, NH 2 , exchanges with D 2 O), 8.12 (s, 1H, C 5 H), 12.30 (b, 1H, N 3 H, exchanges with D 2 O). Anal. Calcd. for C 5 H 4 N 4 OS: C, 35.71: H, 2.40: N, 33.31: S, 19.07. Found: C, 35.50: H, 2.36: N, 33.13: S, 18.79. EXAMPLE 21 7-Amino-4-(2,3,5-tri-O-benzoyl-β-D-ribofuranosyl)thiazolo[4,5-d]pyrimidin-2-one ( 31 ). Compound 30 (460 mg, 2.7 mmol) was glycosylated in the same manner as that used to prepare 6 , requiring HMDS (30 mL), benzoyl-blocked sugar ( 5 : 1.5 g, 3.0 mmol), and TMS-triflate (0.76 mL, 3.9 mmol). The reaction mixture was allowed to stir overnight at room temperature and was then worked up as described for 6 to yield 1.6 g (95%) of 31 isolated as a foam: UV λ max (MeOH) 230 nm (ε ), 310 (): 1 H NMR (DMSO-d 6 ) δ 6.45 (d, J = 2.73 Hz, 1H, C 1 ,H), 7.4-8.0 (m, 15H, benzoyl aromatics), 8.59 (s, 1H, C 5 H), and other sugar protons C 31 H 24 N 4 O 8 S: C, 60.78: H, 3.95: N, 9.15: S, 5.23. Found: EXAMPLE 22 7-Amino-3-(2,3,5-tri-O-benzoyl-β-D-ribofuranosyl)thiazolo[4,5-d]pyrimidin-2-one ( 32 ). Compound 30 (1.22 g, 7.25 mmol) was glycosylated as described for the preparation of 6 , requiring HMDS (35 mL), benzoyl-blocked sugar ( 5 : 4.4 g, 8.7 mmol) and TMS-triflate (2.0 mL, 10.3 mmol). After stirring overnight at room temperature, the reaction mixture was refluxed for 2 days and then worked up in the usual manner. The crude mixture was subjected to flash silica gel column chromatography using a gradient of methylene chloride to methylene chloride-acetone 10:1 (v/v) and yielded two products. The first to elute from the column was assumed by 1 H NMR to be a bis-glycoside which amounted to 660 mg. The second and major product off the column was obtained as a foam and assigned as the desired 3-ribosyl isomer by UV and 1 H NMR: yield 1.04 g (24%): UV λ max (EtOH) 232 nm (ε ), 283 (): 1 H NMR (DMSO-d 6 ) δ 6.34 (t, 1H, C 1 ,H), 7.39-7.98 (m, 17H, benzoyl aromatics and NH 2 , 8.19 (s, 1H, C 5 H), and other sugar protons. Anal. Calcd. for C, 60.78: H, 3.95: N, 9.15: S, 5.23. Found: 3.93: N, 8.13: S, 4.85. EXAMPLE 23 7-Amino-4-β-D-ribofuranosylthiazolo[4,5-d]pyrimidin-2-one ( 33 ). Compound 31 (310 mg, 0.51 mmol) was dissolved in dry methanol (35 mL) and cooled to 5°C. To this solution was added solid sodium methoxide (82 mg, 1.5 mmol) and the solution was stirred at room temperature for 5 h. The mixture was neutralized with Dowex-50 H + resin, filtered and evaporated to dryness. The residue was triturated with ethyl ether and then recrystallized from aqueous ethanol to yield colorless needles: 120 mg, 80%: mp 132-134°C: UV λ max (pH 1) 227 nm (ε 17230), 301 (15750): UV λ max (pH 7, 11) 233 nm (ε 22300), 305 (19100): 1 H NMR (DMSO-d 6 ) δ 5.96 (d, J = 3.51 Hz, 1H, C 1 ,H), 7.75 (b, 2H, NH 2 ), and other sugar protons. Anal. Calcd. for C 10 H 12 N 4 O 5 S.O.2H 2 O: C, 35.71: 9.53. Found: C, 35.45: H, 4.88: N, 16.44: S, 9.50. EXAMPLE 24 7-Amino-3-β-D-ribofuranosylthiazolo[4,5-d]pyrimidin-2-one ( 34 ). Compound 32 (0.76 g, 1.2 mmol) was deblocked in the same manner as described for 31 above using sodium methoxide (200 mg, 3.7 mmol) in dry MeOH (50 mL). The title compound ( 34 ) was obtained (0.12, 32%) after crystallization from water: mp 248-250°C: UV λ max (pH 1) 222 nm (ε 35100), 265 (14300), 290 (11400): UV λ max (pH 7, 11) 215 nm (ε 45000), 262 (13200): 1 H NMR (DMSO-d 6 ) δ 5.91 (d, J = 5.43 Hz, 1H, C 1 ,H), 7.44 (b, 1H, NH 2 ), 8.22 (s, 1H, C 5 H), and other sugar protons. Anal. Calcd. for C 10 H 12 N 4 O 5 S: C, 40.00: H, 4.03: N, 18.66: S, 10.68. Found: C, 39.80: H, 3.99: N, 18.39: S, 10.57. EXAMPLE 25 5-Amino-3-(2-deoxy-3,5-di-O-(p-toluoyl)-β-D-erythropentofuranosyl)thiazolo[4,5-d]pyrimidin-2,7-dione (35) . 5-Aminothiazolo[4,5-d]pyrimidin-2,7-dione (4.3 g, 23.3 mmol) was combined with trimethylsilyl trifluoromethanesulfonate (13.6 mL, 73.4 mmol) in hexamethyldisilazane (70 mL) and the mixture refluxed for three hours after which the excess solvent was removed by vacuum distillation. The resulting solid mass was combined with 1-chloro-2-deoxy-3,5-di-O-(p-toluoyl)-α-D-erythro-pentofuranose (11.8 g, 73 mmol) and the mixture fused at 110°C for 30 minutes. The resulting product was dissolved in EtOAc (400 mL) and poured into a stirring aqueous 5% NaHCO 3 solution. The organic phase was isolated, dried over anhydrous sodium sulfate, and purified by flash column chromatography (dichloromethane/acetone, 4:1 as eluant) to give 1.7g (14%) of a mixture of α and β isomers; mp 268-270; UV λ max (pH 1) 243, 300 nm; UV λ max (pH 7) 243, 302 nm; UV λ max (pH 11) 243, 284 nm; 1 H NMR (Me 2 SO-d6) σ 6.37 (t,1,C 1 'H) 7.0 (bs, 2, NH 2 , exchanged with D 2 0) 7.2 and 7.9 (m, 5, CH 3 C 6 H 5 ) 11.35 (bs, 1, N 6 H, exchanged with D 2 0) and other sugar protons. NMR seems to indicate only one isomer. EXAMPLE 26 5-Amino-3-(2-deoxy-β-D-erythropentofuranosyl)thiazolo[4,5-d]pyrimidine-2,7(3H,6H)-dione (36) . Sodium methoxide (120 mg, 2.2 mmol) was added to a solution of 5-Amino-3-(2-deoxy-3,5-di-O-(p-toluoyl)-β-D-erythropentofuranosyl)thiazolo[4,5-d]pyrimidin-2,7-dione (35; 0.465g, 0.87 mmol) in anhydrous methanol (100 mL) and the resulting solution stirred at room temperature for 8 hours. The solution was neutralized with Dowex 50W-X8 (H + form) resin, filtered, and evaporated to dryness and the residue treated with anhydrous ether (35 mL). After trituration, the suspension was decanted to give a dry powder which was crystallized (charcoal treatment, Norit A) from ethanol. After filtering off the initial precipitate; 110 mg (42%) of the β isomer could be recovered by fractional crystallization using ethanol. mp >170°C (scinters), UV 1 H NMR (Me 2 SO-d 6 ) σ 4.68 (t, 1, J = 5.7 Hz, C 3 'OH, exchanged with D 2 0), 5.20 (d, 1, J = 3.9 Hz, C 5 'OH, exchanged with D 2 0), 6.24 (t, 1, J = 7.2 Hz, C 1 'H), 6.94 (bs, 2, NH 2 , exchanged with D 2 0), 11.23 (bs, 1, N 6 H, exchanged with D 2 0); Anal. Calcd. for C 10 H 12 N 4 O 5 S: C, 40.00; H, 4.03; N, 18.66; S, 10.68; Found: C, 40.27; H, 3.92; N, 18.75; S, 10.42. EXAMPLE 27 5-Amino-3-(β-D-xylofuranosyl)thiazolo[4,5-d]pyrimidine-2,7-(3H,6H)-dione (38) . The title compound is prepared by glycosylation of 4 exactly as described above for the synthesis of 6 except that 1,2,3,5-tetra-O-acetyl-D-xylofuranose is to be used. The crude acetyl-blocked nucleoside (37) is then deprotected using sodium methoxide as described for the preparation of 7 to yield 38 as an amorphous solid. EXAMPLE 28 5-Amino-3-(2,3,5-tri-O-benzyl-β-D-arabinofuranosyl)thiazolo[4,5-d]pyrimidine-2,7(3H,6H)-dione (39) . The title benzyl-protected nucleoside is prepared in the same manner as that described for 35 except that the sugar to be used, 1-chloro-2,3,5-tri-O-benzyl-α-D-arabinofuranose, is generated from the corresponding 1-O-(p-nitrobenzoyl) sugar using dry hydrogen chloride gas in dichloromethane at 0°C. The mixture of anomers is separated by flash silica gel column chromatography to yield 39 and its α anomer, both as dry foams. EXAMPLE 29 5-Amino-3-(β-D-arabinofuranosyl)thiazolo[4,5-d]pyrimidine-2,7(3H,6H)-dione (40) . Compound 39 is dissolved in dry dichloromethane and deprotected using an excess of boron trichloride (1M in dichloromethane) at -78°C and allowed to warm to -40°C for 2 hours before quenching with methanol. The residue after workup is crystallized from water to yield 40 as a colorless crystalline solid. EXAMPLE 30 5-Amino-3-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)thiazolo[4,5-d]-pyrimidine-2,7(3H,6H)-dione (41) . Compound 4 (10 g, 54.4 mmol) was glycosylated exactly as described for the synthesis of 6 except that the sugar used in this case was the 1,2,3,5-tetra-O-acetyl-D-ribofuranose (20.7 g, 65.1 mmol). After the usual workup the crude residue was flash chromatographed on silica gel using 10% methanol in dichloromethane. Yield of pure material 7.1 g (29%) as a dry foam or amorphous solid. EXAMPLE 31 5-amino-3-(2,3-di-O-isopropylidene-β-D-ribofuranosyl)thiazolo-[4,5-d] pyrimidine-2,7-dione (42) . To a ice-cold suspension of 5-amino-3-(β-D-ribofuranosyl)thiazolo[4,5-d]pyrimidine-2,7-dione (7; 1.86 g, 6 mmol) in a mixture of acetone (100 mL) and dimethoxypropane (150 mL) was added perchloric acid (1.0 mL) dropwise, and the resulting mixture stirred at 0° for 30 minutes. The reaction mixture was neutralized in cold to pH 7 with 1N sodium hydroxide. The resulting solution was concentrated in vacuo and the residue flash chromatographed on silica gel with 10% acetone in chloroform as eluent to yield the title compound as an amorphous solid. (1.7g, 80%). 1 H NMR (DMSO d 6 , 300 MHz): δ 1.28, 1.47 (2s, 6H, isopropylidene methyls),6.0 (d, 1H, C 1 H), 7.0 (bs, 2H, NH 2 ), 11.29 (s, 1H, NH) and other sugar protons. Anal. Calcd. for C 13 H 16 N 4 SO 6 : C, 43.82; H, 4.53; N, 15.72; S, 9.00. Found: C, 43.65; H, 4.48; N, 15.41; S, 8.94. Natural killer cells have been implicated as providing defense against viral infections and malignant cells. Abnormalities in the natural killer cell's activity may thus result in the development of the diseases. Biological immunomodulators may restore or correct certain deficient immune functions. Recently interleukin-2 has been shown to have immunotherapeutic potential in tumor patients. Interleukin-2 and other known immunopotentiators are generally protein in nature and may cause severe side effects upon their administration. Non toxic low molecular synthetic compounds which are not proteins can be suggested for avoiding the side effects of known protein immunotherapeutic potentiators. EXAMPLE 32 - In Vitro Induced Potentiation of Natural Killer Cell Activity The non protein nucleoside and nucleotide compounds used in the invention have shown increased natural killer cell activity. In this example, natural killer cell activity is demonstrated in mice. Spleen cells from CBA/CaJ or C57BL/6J mice were cultured with 0.05 mM concentrations of compound 7 for 20 to 44 hours at 37°C in a 5% CO 2 humidified atmosphere as described in Djeu, J-Y, Heinbaugh, J. A., Holden, H. T., Herberman R. B.: Journal Immunology 122:175, 1979, and Gonzales, H., Karriger, K., Sharma, B., Vaziri, N.: Federal Procedures 42:1195, 1983. After incubation the cytotoxicity of the treated and untreated cells was determined against YAC-1 cells. In performing this test both a non-drug control and a control using Poly I:C were run concurrently with compound 7 . In Vitro Induced Potentiation of Natural Killer Cell Activity Effector Cells % Natural Killer Cell Cytotoxicity From: Treatment With: Treatment Time (Hour) 0 20 44 CBA/CAJ None 13 4 0.5 Compound 7 15 34 31 Poly I:C 14.5 18 19 CBA/CAJ None 14 8 1 Compound 7 23 62 44 Poly I:C 22 31 9 C57BL/6J None 16 1.5 2.5 Compound 7 18 35 34 Poly I:C 18 13 14 C57BL/6J None 30 1.6 1.3 Compound 7 18 25 22 Poly I: 22 6 13 As is evident from Table 1 incubation for 20 hours with compound 7 augmented natural killer cell cytotoxicity from 4, 8, 1.5 and 1.6% for untreated control to 34, 62, 35 and 25% respectively. Similar treatment for up to 44 hours also caused a distinct increase in natural killer cell activity. Using Poly I:C as a further control, which is a well known potentiator of natural killer cells, compound 7 demonstrated increased activity compared to Poly I:C. The results of Table 1 demonstrate compound 7 markedly induces a high increase in murine natural killer cell activity. EXAMPLE 33 - In Vitro Augmentation of Human Natural Killer Cell Activity The natural killer cell activity in human cells was also measured. In conducting these tests peripheral blood mononuculear cells (PBMNC) were isolated on Ficoll-Hypaque gradient, washed three times in Hanks and re-suspended in RPMI-1640 containing 10% human AB serum as described in Rosenberg, S.A.: Journal American Medical Association , 256:3117, 1986 and the above referred to reference to Djeu, et al. Peripheral blood mononuclear cells (PBMNC) from eleven different donors were treated with compound 7 as described in Djeu, et al. as well as Sharma, B., Odom, L. F.: Cancer Immunol. Immunother. 7:93, 1979, and Sharma, B., Odom, L. F.: Cancer Research 44:3258, 1984. The PBMNC cells were incubated with compound 7 at 37°C for 20 to 68 hours in a 5% CO 2 humid atmosphere and after incubation the cytotoxicity was determined against K562 tumor cells. The results are shown in Tables 2a and 2b. TABLE 2a In Vitro Augmentation of Human Natural Killer Cell Activity Donor % Natural Killer Cell Cytotoxicity - Compound 7 + Compound 7 1 26 91 2 4 29 3 22 54 4 24 47 5 6 16 6 1 18 7 2 10 8 13 22 9 6 15 10 14 20 11 11 32 In Vitro Augmentation of Human Natural Killer Cell Activity Exp. # % Natural Killer Cell Cytotoxicity Donor 1 Donor 2 -Compound 7 +Compound 7 -Compound 7 +Compound 7 1 25 47 4 29 2 26 91 15 37 3 9 36 19 37 4 23 55 3 26 5 32 67 4 13 6 12 30 2 10 7 16 50 8 31 40 As evident from Table 2a the PBMNC from eleven donors had a mean natural killer cell cytotoxicity activity of 11.7%. The PBMNC from the same donors which was pretreated with compound 7 expressed a 32.2% mean natural killer cell cytotoxicity. While there is some variability between the eleven donors, eight of the donors showed over 100% potentiation in natural killer cell cytotoxicity. In the other four cases compound 7 treatment in vitro produced a marked increase in natural killer cell mediated cytotoxicity. As is seen in Table 2b compound 7 also consistently mediated potentiation of human natural killer cells. In the first donor eight individual tests were run and an increase in the mediated potentiation was seen in eight of the eight tests. In the second donor the increase was seen in six of six tests. As is evident from Tables 2a and 2b compound 7 significantly induced higher levels of natural killer cell activity in human cells. In further reproducibility tests compound 7 was tested for reproducibility in its increase in natural killer cell activity as is shown in Table 2c and further for its reproducability on human natural killer cell activity as is shown in Table 2d. Reproducibility of Compound 7 Induced Increase in Natural Killer Cell Activity Exp. # % Increase in Natural Killer Cell Activity Donor 1 3 1 1031 220 2 658 102 3 413 201 4 200 66 5 149 28 6 460 136 7 91 102 8 98 200 9 200 Effect of Compound 7 on Human Natural Killer Cell Activity Donor % Increase in Induced Natural Killer Cell Activity 1 1031 2 548 3 220 4 88 5 123 6 141 7 167 8 150 9 0 10 63 11 8 EXAMPLE 34 - In Vivo Potentiation of Natural Killer Cell Activity in Mice Compound 7 was further studied for potentiation of natural killer cell activity in vivo in mice. CBA/CaJ mice were treated with compound 7 by injecting 1.68 mg per 0.5 ml per mouse of compound 7 . After 1, 2, 3 and 4 days of treatment the spleens of the mice were harvested and the cytotoxic activity of the spleen cells was determined against YAC-1 tumor target cells in the above referred to 51 Cr release assay as identified in EXAMPLE 32. The results of these tests are shown in Table 3, 4, 5 and 6. Natural Killer cell Activity in CBA/CaJ Mice Mice CBA/CaJ Treatment With: Days After Treatment % Natural Killer Cytotoxicity Effector: Target 50:1 100:1 150:1 Group - 1 Saline 1 11 ± 3 18 ± 2 23 ± 0.3 Group - 2 Compound 7 (1.68mg) 1 53 ± 5 64 ± 5 71 ± 9 Group - 3 Compound 7 (1.68mg) 2 46 ± 9 64 ± 5 65 ± 7 Group - 4 Compound 7 (1.68mg) 3 38 ± 6.6 50 ± 7 63 ± 6 Group - 5 Compound 7 (1.68mg) 4 30 ± 0.1 37 ± 1.5 48 ± 3 As is evident from table 3 a single injection of compound 7 at 1.68 mg per mouse caused a profound increase in natural killer cell activity. A maximum response of 382% increase was obtained one day after treatment. More than a two fold increase was observed even after 4 days with compound 7 . EXAMPLE 35 - Dosage Effects on Natural Killer Cell Activity in Mice Table 4 shows a dose response treatment with compound 7 . Dosage Effects on Natural Killer Cell Activity in Mice Mice CBA/CaJ Treatment With: % Natural Killer Cytoxicity Effector : Target 50:1 100:1 150:1 Group - 1 Saline 14 ± 4 20 ± 4 23 ± 3.5 Group - 2 Compound 7 (0.84mg) 38 ± 7 48 ± 9 56 ± 8.5 Group - 3 Compound 7 (1.68mg) 44 ± 8 56 ± 12 61 ± 14 Group - 4 Compound 7 (2.52mg) 48 ± 6.6 62 ± 7 70 ± 5 Group - 5 Compound 7 (3.36mg) 48 ± 4.6 67 ± 10 74 ± 6.4 As is evident from Table 4 mice were treated with 0.84 mg to 3.36 mg per mouse of compound 7 . Although all doses of compound 7 induced a marked increase in natural killer cell activity the maximum augmentation was displayed by mice that received 3.36 mg of compound 7 . EXAMPLE 36 - Natural Killer Cell Activity in Old Mice Twelve week old C57BL/6J mice have been shown (Djeu et al. above) to exhibit little spontaneous natural killer cell activity. Table 5 shows the effect of increase of natural killer cell activity in aging mice (8 Months Old). For this test compound 7 was injected at a dose of 1.67 mg per mouse and 3.34 mg per mouse and after a day the natural killer cell activity of the spleen cells was determined. Natural Killer Cell Activity in Old Mice Mice C57BL/6J Treatment With: %Natural Killer Cell Cytotoxicity Effector: Target 50:1 100:1 150:1 Group - 1 (8 Mo. Old) Saline 4 9 14 Group - 2 (8 Mo. Old) Compound 7 (1.67mg) 14 19 23 Group - 3 (8 Mo. Old) Compound 7 (3.34mg) 17 28 38 Group - 4 (8 Wk. Old) Saline 5 7 7 Group - 5 (8 Wk. Old) Compound 7 (1.67mg) 15 20 24 Group - 6 (8 Wk. Old) Compound 7 (3.34mg) 17 27 31 As is evident compound 7 showed an increase in natural killer cell activity ranging from 4% untreated to 17% treated at a 50:1 effector/target ratio. The magnitude of induction of increase was similar to that displayed by compound 7 in 8 week old mice. This low molecular weight nucleoside compound induced increases in natural killer cell activity which is as high as increases mediated by Poly I:C, LPS, Pyran or interferon as has been reported by Djeu et al above. EXAMPLE 37 - Natural Killer Cell Activity in Nude/Nude Mice Compound 7 was also able to potentiate markedly the natural killer cell activity in T cell deficient (nude/nude) mice as is shown in Table 6. Natural Killer Cell Activity in Nude/Nude Mice Mice Nude-Nude Treatment With: % Natural Killer Cell Cytotoxicity Effector : Target 50:1 100:1 150:1 Group - 1 Saline 15 ± 5 25 ± 7.7 30 ± 11 Group - 2 Compound 7 (1.67mg) 43 ± 12 56 ± 12 61 ± 14 Group - 3 Compound 7 (3.34mg) 38 ± 6 56 ± 6 61 ± 12 As is evident from table 6 compound 7 showed an effective increase of over two fold at both doses tested with respect to a saline control. Compounds of the invention have also been tested for antiviral activity. Tests have been conducted for both the therapeutic and prophylactic effect of the compounds against a variety of both RNA and DNA viruses. EXAMPLE 38 - Antiviral Activity Against Herpes Simplex Virus Types 1 and 2 In this test Compound 7 was tested against both Herpes Simplex Type 1 Virus and Herpes Simplex Type 2 Virus. The tests were conducted as prophylactic - treatments in vivo utilizing the mouse as a model. In each of these tests a placebo was utilized for control purposes. The survival time reflects survival time of those animals which succumbed during the test. Antiviral Activity Against Effects of Herpes Simplex Virus Type 1 Infection Compound Dose (mg/kg/day) Survivors/Total (%) Mean Survival Time (days) Placebo - 1/12 (0) 12.5 ± 3.0 Compound 7 200 6/12 (50) 11.0 ± 1.7 Compound 7 100 4/12 (25) 11.4 ± 1.8 Compound 7 50 8/12 (75) 14.8 ± 5.0 As is shown in Table 7 prophylactic treatment of a Herpes type 1 infection in mice was effectively treated with compound 7 . There was some variability in the response with the response at 100 mg per kg dose being less effective than both the lower and higher dosages. Antiviral Activity Against Herpes Simplex Virus Type 2 Infection Compound Dose (mg/kg/day) Survivors/Total (%) Mean Survival Time (days) Placebo - 0/12 (8) 9.8 ± 1.0 Compound 7 200 2/12 (17) 7.2 ± 3.9 Compound 7 100 6/12 (50) 11.2 ± 1.5 Compound 7 50 6/12 (50) 11.8 ± 2.3 As is shown in Table 8 prophylactic treatment of a Herpes type 2 infection was effective at all levels tested. At 50 and 100 mg per kg this statistically significance in mean survival time was just outside of the range of a statistical significance of p<0.1 by the two-tailed Fisher exact test. At 200 mg per kg in the mouse the dose was partially toxic. EXAMPLE 39 - Antiviral Activity Against Influenza B Virus Infection in Mice Compound 7 was tested therapeutically against Influenza B Virus Infection in Mice. In addition to a saline control Ribavirin, a known antiviral, was utilized for test purposes. The results of this test are shown in Table 9. Antiviral Activity Against Influenza B Virus Infection in Mice Compound Dose (mg/kg/day) Treatment Schedule Survivors/Total (%) Mean Survival Time (days) Saline - a 0/12 (0) 8.7 ± 2.7 Ribavirin 100 a 10/12 (83) 8.0 ± 0.0 Compound 7 100 b 2/12 (17) 6.6 ± 1.1 50 b 2/12 (17) 6.4 ± 1.5 a Half-daily doses were administered twice a day for 7 days starting 2 hours pre-virus inoculation. b Treated once a day at -2 hours and on days 2, 4, and 6 relative to virus inoculation. As is shown in Table 9 against influenza B in the mouse compound 7 has an efficacy between saline, having no antiviral activity against influenza, and Ribavirin, which has significant antiviral activity against influence. EXAMPLE 40 - Antiviral Activity Against San Angelo Virus Infection in Mice. In this example the antiviral activity against a San Angelo virus, an encephalitis type virus, was measured utilizing both a therapeutic and a prophylatic protocol. The results of this test are given in Table 10. As with the prior example Ribavirin was utilized as a positive antiviral control and saline as a negative. The results of this test is given in table 10. Antiviral Activity Against San Angelo Virus Infection in Mice Compound Dose (mg/kg/day) Treatment Schedule Survivors/Total (%) Mean Survival Time (21 days) Saline - a 2/12 (17) 7.5 ± 1.2 Ribavirin 200 a 11/12 (92) 12.3 ± 2.9 Compound 7 200 b 12/12 (100) >21 100 c 12/12 (100) >21 50 c 12/12 (100) >13.0 ± 0.0 Compound 7 200 d 12/12 (100) >21 a Half-daily doses were administered twice a day for 7 days starting 2 hours pre-virus inoculation. b Treated once a day at -2 hours and on days 2, 4, and 6 relative to virus inoculation. Half daily doses were administered twice a day. c Treated once a day at -2 hours and on days 2, 4, and 6 relative to virus inoculation. d Half-daily doses were administered at 24 and 16 hours pre-virus inoculation. As is evident from Table 10 in both a therapeutic and a prophylactic mode compound 7 showed antiviral activity equal to that of Ribavirin against San Angelo encephalitis virus. EXAMPLE 41 - Antiviral Activity Against Mouse Cytomegalovirus Infection in Mice. In this test compound 7 was also tested for both therapeutic or prophylactic efficacy against a mouse cytomegalovirus infection. The results of these tests are shown in Table 11 and 12. Antiviral Activity Against Mouse Cytomegalovirus Infection in Mice Compound Dose (mg/kg/day) Treatment Schedule Survivors/Total (%) Mean Survival Time (days) Saline - a 4/12 (33) 6.1 ± 0.4 Compound 7 200 a 12/12 (100) >21 100 b 7/12 (58) 7.0 ± 1.4 50 b 6/12 (50) 6.5 ± 0.8 a Half-daily doses were administered at 24 and 16 hours previrus inoculation b Single dose was administered 24 hours pre-virus inoculation Antiviral Activity Against Mouse Cytomegalovirus Infection in Mice Compound Dose (mg/kg/day) Treatment Schedule Survivors/Total (%) Mean Survival Time (days) Saline - a 3/12 (25) 6.1 ± 0.9 Compound 7 100 b 0/12 (0) 6.0 ± 0.8 50 b 0/12 (0) 6.3 ± 1.1 a Treatments were once a day for 6 days starting 2 hours pre-virus inoculation b Treatments were once a day at -2 hours and on days 2, 4, and 6 relative to virus inoculation As is evident at the 200 mg per kg dose compound 7 exhibited a 100% cure when tested in a prophylactic mode. However, as is seen in Table 12 this activity was not repeated in a therapeutic mode. EXAMPLE 42 - Antiviral Activity Against Semliki Forest Virus Infection in Mice. Antiviral activity was also tested against Semliki Forest Virus an Encephalitis type virus. In this example compounds 7 and 36 were tested for therapeutic efficacy and additionally compound 7 was tested for prophylactic efficacy. Results of this test are shown in Table 13 for the therapeutic mode and in Table 14 for the prophylactic mode. Therapeutic Antiviral Activity Against Semliki Forest Virus Infection in Mice Compound Dose (mg/kg/day) Treatment Schedule Survivors/Total (%) Mean Survival Time (days) Saline - a 0/12 (0) 6.7 ± 1.9 Compound 7 200 b 7/12 (58) 4.2 ± 0.8 100 c 8/12 (67) 7.3 ± 0.5 50 c 4/12 (33) 6.6 ± 0.8 Compound 36 200 10/12 (83) 7.0 ± 0.0 100 6/12 (50) 6.8 ± 0.8 50 4/12 (33) 6.1 ± 1.1 a Half-daily doses were administered twice a day for 7 days starting 2 hours pre-virus inoculation b Treated at -2 hours and on days 2, 4, and 6 relative to virus inoculation. Half-daily doses were administered twice a day because of insolubility c Treated once a day at -2 hours and on days 2, 4, and 6 relative to virus inoculation d Half-daily doses were administered at 24 and 16 hours pre-virus inoculation Prophylactic Antiviral Activity Against Semliki Forest Virus Infection in Mice Compound Dose (mg/kg/day) Treatment Schedule Survivors/Total (%) Mean Survival Time (days) Saline - d 1/8 (12) 10.4 ± 1.3 Compound 7 200 d 4/8 (50) 11.8 ± 2.9 a Half-daily doses were administered twice a day for 7 days starting 2 hours pre-virus inoculation b Treated at -2 hours and on days 2, 4, and 6 relative to virus inoculation. Half-daily doses were administered twice a day because of insolubility c Treated once a day at -2 hours and on days 2, 4, and 6 relative to virus inoculation d Half-daily doses were administered at 24 and 16 hours pre-virus inoculation e p<.01 two-tailed Fisher As is evident from Tables 13 and 14 compound 7 exhibited antiviral activity in both a therapeutic mode and a prophylactic mode against this virus. Further compound 36 also exhibited therapeutic antiviral activity against this virus. As is evident from the above tables antiviral activity against a variety of viruses is seen. In a further test little or no antiviral activity for compound 7 was demonstrated against influenza B virus and Friend Leukemia virus induced spleenomegaly. EXAMPLE 43 - Antiviral Activity Against Human Coronavirus In this experiment, 14 day old mice were inoculated intracerebrally with human coronavirus. They were treated with compound 7 in a divided intraperitoneal dose at -24 and -18 hours relative to virus inoculation. As shown in the following Table 15, a statistically significant benefit against this viral infection in the brain was provided to the mice at 200 and 100 mg/kg. Antiviral Activity Against Human Coronavirus Infection In 14 Day Old Mice Test Compound (mg/kg) Survivors/Total (%) Mean Survival Time (Days) 0 0/33 (0) 6.0 ± 1.6 200 4/32 (13) 7.0 ± 1.4 100 8/33 (24) 7.2 ± 2.0 50 0/33 (0) 6.9 ± 2.2 As is evident from table 15 compound 7 shows antiviral activity against this human virus which was inoculated into the brain. EXAMPLE 44 - Antiviral Activity Against Rat Coronavirus Four to five day old suckling rats were infected intranasally with rat coronavirus. One day previously a divided intraperitoneal dose of compound 7 or placebo was given. As shown in the following Table 16, a high percentage of the rats treated with compound 7 survived this lung infection. Further the rats that died lived longer than the placebo-treated controls. Antiviral Activity Against Rat Coronavirus Infection In Suckling Rats Test Compound (µM) Survivors/Total (%) Mean Survival Time (Days) 0 7/28 (25) 7.2 ± 1.8 200 22/27 (81) 13.0 ± 1.0 100 17/28 (61) 10.2 ± 1.8 50 21/28 (75) 10.1 ± 2.3 EXAMPLE 45 - Antiviral Activity Against Encephalomyocarditis Viral Infection In Mice In this experiment, the effects of different treatment regimens of compound 7 on the outcome of an Encephalomyocarditis viral infection in mice was studied. Treatments were started 24 hours pre-virus. A single dose of 200 mg/kg was given to two groups of mice. One of the groups then received single injections of compound 7 at 50 mg/kg for 6 more days. Results were compared to a placebo control treated as was the 50 mg/kg group. As shown in the following Table 17 there is clear evidence that mice receiving the daily dose of 50 mg/kg were protected to a greater degree than the mice which only got the single 200 mg/kg dose. The former group had less mortality and extended means survival times. Effects of single versus multiple treatments on Encephalomyocarditis virus infection in mice Compound Treatment -24 hrs Treatment Days 0-6 Survivors/Total (%) Mean Survival Time (Days) Placebo + + 1/12 (8) 4.4 ± 0.5 Test Compound 200 - 5/12 (42) 6.0 ± 1.3 Test Compound 200 50 8/12 (67) 9.5 ± 2.1 EXAMPLE 46 - Antiviral Activity of Nucleosides and Nucleotides on Murine Natural Killer Cell Activity In Vitro Other compounds used in the invention were tested with respect to their natural killer cell activity utilizing mouse spleen cells as per example 32 above. The results of these tests arc tabulated in Table 18. Effect of Guanosine Nucleosides and Nucleotides on Murine Natural Killer Cell Activity In Vitro Exp # Effector Cells Pretreated with: Concentration (mM) % Natural Killer Cell Cytotoxicity 1 None - 1.2 Compound 7 0.05 34.5 Compound 16 0.05 10 Compound 16 0.5 38 Compound 9 0.05 3 Compound 9 0.5 13 2 None - 1.6 Compound 7 0.05 25 Compound 7 0.25 28 Compound 16 0.05 3 Compound 16 0.25 17 Compound 9 0.05 2 3 None - 1.6 Compound 7 0.05 28 Compound 16 0.05 14 Compound 16 0.25 24 Compound 9 0.05 0.31 Compound 9 0.5 6.5 EXAMPLE 47 - Effect of Nucleosides and Nucleotides on Human Natural Killer Cell Activity In Vitro Other Nucleosides and Nucleotides used in the invention were tested for their activity in human natural killer cells in vivo as per example 33 above. The results are tabulated in Table 19. Effect of Guanosine Nucleosides and Nucleotides on Human Natural Killer Cell Activity In Vitro Exp # Effector Cells Pretreated with: Concentration (mM) % Natural Killer Cell Cytotoxicity 1 None - 26 Compound 7 0.4 91 Compound 16 0.4 69 Compound 12 0.4 39 Compound 11 0.4 63 2 None - 23 Compound 12 0.2 43 3 None - 17 Compound 12 0.4 22 4 None - 9 Compound 11 0.4 19 5 None - 3 Compound 7 0.4 37 Compound 16 0.4 10 Compound 9 0.05 6 6 None - 16.53 Compound 7 0.4 50 None - 40 Compound 9 0.2 43 7 None - 4.5 Compound 7 0.2 10 Compound 8 0.2 5 Compound 19 0.2 7 In examples 48 and 49 the combination activity of compounds of the invention with the known antiviral agent Ribavirin against San Angelo Virus and Banzi Virus were measured utilizing compound 7 in a prophylatic protocol. The Ribavirin served as a further antiviral agent for use in combination with the compounds used in the invention. EXAMPLE 48 - Combination Chemotherapy Against San Angelo Virus In Mice The combination chemotherapy results against San Angelo Virus in Mice are shown in Table 20. Combination Chemotherapy Against San Angelo Virus Treatment Survivors/Total(%) Mean Survival Time (Days) -24 Hrs Days 0-6 Saline Saline 1/16 (6) 8.3 ± 1.8 Compound 7 (5) Saline 7/16 (44) 9.6 ± 1.6 Saline Ribavirin (50) 6/16 (38) 9.0 ± 1.2 Saline Ribavirin (25) 9/16 (56) 8.1 ± 0.7 Compound 7 (5) Ribavirin (50) 8/16 (50) 8.9 ± 2.2 Compound 7 (5) Ribavirin (25) 6/16 (38) 10.1 ± 3.6 EXAMPLE 49 - Combination Chemotherapy Against Banzi Virus Infection In Mice The combination chemotherapy results against Banzi Virus infection in mice are shown in Table 21. Combination Chemotherapy Against Banzi Virus Infection Treatment -24 hours Treatment days 0-6 Survivors/Total (%) Mean Survival Time (days) Placebo Saline 0/12 (0) 7.7 ± 0.5 Placebo Ribavirin 100 mg/kg 0/12 (0) 8.5 ± 1.0 Placebo Ribavirin 200 mg/kg 0/12 (0) 9.1 ± 0.8 Compound 7 100 mg/kg Saline 0/12 (0) 9.3 ± 1.0 Compound 7 100 mg/kg Ribavirin 100 mg/kg 0/12 (0) 10.2 ± 0.9 Compound 7 Ribavirin 3/12 (25) 12.4 ± 3.5 As is evident from Tables 20 and 21, compound 7 in a prophylactic mode in combination with the known antiviral Ribavirin, exhibited efficacy against the test viruses. The compounds used in the invention can be given to a warm blooded host in need thereof in appropriate formulations wherein the compounds comprise the active ingredient of the formulations. Thus the compounds of the invention can be made up into injectables suitable for intravenous or other type injection into the host animal. Further they can be given in an appropriate oral formulation as for instance as an oral syrup preparation, an oral capsule or oral tablet. An additional route of administration might be as a suppository. For an injectable the compounds would be dissolved in a suitable solution as for instance in a sodium bicarbonate or other buffer. Such a solution would be filtered and added to appropriate ampoules or vials and sealed and sterilized. As a syrup, the compounds in buffered solution would be mixed with an appropriate syrup with mild stirring. For capsules the dry compounds would be blended with appropriate fillers, binders or the like as for instance Lactose USP powder or Sterotex powder. For the preparation of tablets the compounds of the invention would be mixed with suitable binders and fillers as for instance corn starch NF, Microcrystalline Cellulose, Sterotex powder and water and dried to a low water content. This would be followed by screening, milling, further screening and pressing into the appropriate tablets. For suppositories, the compounds would be dissolved into appropriate melts of Polyethylene Glycol as for instance Polyethylene Glycol 1540 and 8000 at 60° and formed into the suppositories by molding at 25°. In addition to the above formulations, the compounds of the invention could also be administered utilizing other delivery technique such as incorporating the compounds of the invention with liposomes and the like. Additionally, prodrug forms of the compounds of the invention could be utilized to facilitate dispensing, uptake, absorption, metabolic control and the like. One such prodrug is compound 41 , the tri-acetate ester of compound 7 . Further prodrugs might allow for enzymatic conversion in vivo of analogs of the compounds used in the invention into compounds used in the invention. For the purpose of brevity in certain chemical figures and schemes of this specification and the claims attached hereto, different tautomeric forms of the heterocycles of certain compounds have been shown between the various figures and schemes. It is understood that regardless of whether or not substituents are shown in their enol or their keto form, they represent the same compound. Thus, in the claims, the abstract and the brief description in order to utilize only a single structural figure, oxygen and sulfur substituents in the 5 and 7 ring positions are shown in an enolate form whereas in the various schemes these substituents are shown in their normal keto form.

Claims: Use of a compound of the structure: wherein R 4 , R 5 , R 6 and R 7 individually are H, OH or C 1 -C 18 O-acyl and R 3 is H, C 1 -C 18 acyl or or R 5 and R 7 are H or OH, R 6 is H and together R 3 and R 4 are and X is =O or =S; Y is -OH, -SH, -NH 2 or halogen; and Z is H, -NH 2 , -OH or halogen; wherein halogen is Cl or Br, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for use in treating a virus in a mammal. Use according to claim 1 wherein said compound is a compound of the structure: wherein R 1 and R 2 individually are H or C 1 -C 18 acyl and R 3 is H, C 1 -C 18 acyl or or R 1 is H and together R 2 and R 3 are and X is =O or =S; Y is -OH, -SH, -NH 2 or halogen; and Z is H, -NH 2 , -OH or halogen; wherein halogen is Cl or Br; or a pharmaceutically acceptable salt thereof. Use according to claim 1 or 2 wherein: Z is -NH 2 and Y is -OH. Use according to claim 2 or 3 wherein: R 1 and R 2 are H, acetyl or benzoyl and R 3 is H, acetyl, benzoyl or or R 1 is H and together R 2 and R 3 are or a pharmaceutically acceptable salt thereof. Use according to claim 4 wherein: Z is -NH 2 and Y is -OH. Use according to claim 5 wherein: X is =O. Use according to any one of clams 4 to 6 wherein R 1 and R 2 are H. Use according to claim 7 wherein R 3 is H. Use according to any one of claims 1 to 8 wherein the virus is an RNA virus or a DNA virus. Use according to claim 9 wherein the virus is Herpes Simplex Type 1 Virus or Herpes Simplex Type 2 Virus. An anti viral composition containing a therapeutically effective amount of a compound of the structure: wherein R 1 and R 2 individually are H or C 1 -C 18 acyl and R 1 is H, C 1 -C 18 acyl or or R 1 is H and together R 2 and R 3 are and X is =O or =S; Y is -OH, -SH, -NH 2 or halogen; and Z is H, -NH 2 , -OH or halogen; wherein halogen is Cl or Br; or a pharmaceutically acceptable salt thereof in combination with a therapeutically acceptable diluent or carrier. A composition according to claim 11 wherein: Z is -NH 2 and Y is -OH. A composition according to claim 12 wherein: R 1 and R 2 are H, acetyl or benzoyl and R 3 is H, acetyl, benzoyl or or R 1 is H and together R 2 and R 3 are or a pharmaceutically acceptable salt thereof. A composition according to claim 13 wherein: Z is -NH 2 and Y is -OH. A composition according to claim 14 wherein: X is =O. A composition according to claims 13, 14 or 15 wherein: R 1 and R 2 are H. A composition according to claim 16 wherein: R 3 is H. A composition according to claim 11 wherein the compound is 5-amino-3-β- D -ribofuranosylthioazolo[4,5- d ] pyrimidine-2,7(6H)-dione or a pharmaceutically acceptable salt thereof. A composition according to claim 18 wherein the compound is in the form of the 5'-phosphate. A composition according to claim 18 wherein the compound is in the form of the 3'-5' cyclic phosphate. An anti-viral composition containing a therapeutically effective amount of 5-amino-2-thioxo-3-β- D -ribofuranosylthioazolo[4,5- d ]pyrimidin-7(6H)-one in combination with a therapeutically acceptable diluent or carrier. An anti-viral composition containing a therapeutically effective amount of 5-amino-3-β- D -ribofuranosylthiazolo[4,5- d ]pyrimidine-2,7(6H)-dione or a pharmaceutically acceptable salt thereof in combination with a therapeutically acceptable diluent or carrier. An anti-viral composition according to any one of claims 11 to 22 further comprising a second anti-viral agent.

Inventors: COTTAM HOWARD BRINKERHOFF, ROBINS ROLAND KENITH, COTTAM, HOWARD BRINKERHOFF, ROBINS, ROLAND KENITH
Applicants: ICN PHARMACEUTICALS, ICN PHARMACEUTICALS, INC.
Application year: 1988
Main classification: A61K31
IPC codes: A61K31, C07H19