DD251984A5 - PROCESS FOR THE PREPARATION OF 3'-AZIDONUCLEOSIDES AND PHARMACEUTICALLY CONFIDENT DERIVATIVES THEREOF - Google Patents

Abstract

The invention relates to a process for the preparation of novel 3-azidonucleosides and pharmaceutically acceptable derivatives thereof which are suitable in therapy, in particular in the treatment and prophylaxis of Gram-negative bacterial infections and retroviral infections, in particular of AIDS. For example, 3-azido-5-chloro-2,3-dideoxyuridine or 3-azido-3,5-dideoxy-5- ((N, N-didemethyltiocarbomoyl) thio) thymidine are prepared.

Description

Field of application of the invention

The invention relates to a process for the preparation of 3'-azidonucleosides with pharmaceutically acceptable derivatives thereof.

Characteristic of the known technical solutions

In the field of antiviral chemotherapy, due to the difficulty of attacking the virus while the uninfected host cells remain unaffected, few drugs effectively combat the virus as such. It has long been suspected that in the extremely parasitic nature of viruses, all the necessary facilities for viral replication are provided by the host cell. It has recently been discovered that certain stages in the life cycle of the virus which vary from species to species are specified by the virus itself, and these stages may prove to be susceptible to attack where they deviate sufficiently from any corresponding host cell function , However, due to the large similarities between virus and host functions, effective treatments have proven to be very difficult to identify.

For this reason, compounds identified as being suitable for the treatment of viral infections usually have some toxicity to the host. Therefore, the ideal medication is nontoxic at antivirally effective concentrations, but in the absence of such treatment, the compound should have a good therapeutic ratio, that is, the concentrations at which the treatment is toxic are significantly higher than those at which antiviral Activity is observed. -

One group of viruses that have recently gained particular importance are the retroviruses. The retroviruses are a subset of the RNA viruses that must first "reverse-translate" the RNA of their genome into the DNA for replication ("back translation" or "transcription" commonly describes the synthesis of RNA from DNA.) The viral genome can, as soon as It is in the form of the DNA, incorporated into the host cell genome, whereby the virus can enjoy the full advantage of the transcription / translation machinery of the host cells for the purpose of replication The viral DNA after its incorporation is in principle of the DNA of the host is indistinguishable and the virus can persist in this state as long as the cell is alive Since the virus in this form is in principle unassailable, any treatment must be at a different stage of the life cycle of the virus and it will necessarily continue until all cells infected by the virus have died.

HTLV-I and HTLV-II are both retroviruses and are known to be the causative agents of human leukemia. HTLV-I infections are particularly prevalent and responsible for many deaths worldwide each year. A retrovirus species has also been reproducibly isolated from patients with AIDS. Although comprehensively labeled, there is still considerable controversy over an internationally recognized name for the virus. This is commonly known as either human lymphotropic T-cell virus III (HTLVIII) as AIDS-accompanying retrovirus (ARV) or as lymphadenopathy-accompanying virus (LAV), however, the internationally recognized name of human immunodeficiency virus (human immunodeficiency virus = HIV) will be. This virus (referred to herein as HIV) has been shown to preferentially infect and destroy T cells bearing the OKT4 surface marker and is now generally accepted as the etiological factor of AIDS. The patient gradually loses this set of T-cells, disrupting the overall balance of the immune system, reducing its ability to fight other infections, and being receptive to opportunistic infections that are often fatal. Therefore, the common cause of death in AIDS victims is an opportunistic infection such as pneumonia or carcinomas, which may be virally induced, and not necessarily a direct result of HIV infection. Other conditions associated with HIV infection include thrombocytopenic purpura and Kaposki's sarcoma.

Recently, HIV has also been obtained from other tissue types, including the T4 marker, major B cells, macrophages, and non-blood-accompanying tissue in the central nervous system. This central nervous system infection is not necessarily associated with classic AIDS and has been found in patients with asymptomatic HIV infections. HIV infection of the central nervous system (CNS) is associated with progressive demyelination, which leads to premature deterioration

and associated with such symptoms as encephalopathy, progressive dysarthria, ataxia and disorientation. Other conditions associated with HIV infection include the asymptomatic persistant condition, progressive generalized lymphadenopathy (PGL), and the AIDS-related complex (ARC).

It is now considered that certain chronic neurological infections are caused by retroviruses. Such infections include, for example, multiple sclerosis in humans and goatarthritis encephalitis viral infections in goats and Visna Maedi infections in sheep.

Reports have described the study of compounds against various retroviruses, for example, the Friend leukemia virus (FLV), a mouse or rat virus. For example, Krieg et al. (Exp. Cell Res., 116, [1978] 21-29) that 3'-azido-3'-deoxythymidine is active against FLV in vitro experiments and Ostertag et al. Proc. Nat. Acad., [1974] 71, 4980-85) found that, on the basis of antiviral activity relative to FLV and a lack of cellular toxicity, 3'-azido-3'-dideoxythymidine "bromodeoxyuridine However, De Clerq et al. (Biochem. Pharm., 1980, 29, 1849-1851) found six years later that 3'-azido-3 'could be substituted for the medical treatment of diseases caused by DNA viruses. dideoxythymidine had no appreciable activity against any of the viruses used in their experiments, including such DNA viruses as vaccinia, HSVI and varicella zoster virus (VZV).

Bacteria also pose a problem in therapy, since all living organisms use nearly the same life processes as the others, so that a substance that is toxic to one is also likely to be one. other than toxic. In addition, experience has shown that, over time, strains of bacteria develop that are resistant to the commonly used antibacterial agents.

Object of the invention

The invention was based on the object to use new 3'-Azidonucleoside and pharmaceutically acceptable derivatives thereof.

Explanation of the essence of the invention

It has now been discovered that certain 3'-azidonucleosides as described above are useful in the therapy of viral and bacterial infections, particularly retroviral infections, including HIV infections and Gram-negative bacterial infections, including certain strains of Gram-negative bacteria used antibacterial agents are resistant

 Accordingly, according to the present invention, a process for the preparation of a compound of formula

(D

for use in human or veterinary therapy, wherein A is a purine or pyrimidine base attached at the 9 or 1 position, other than thymine, or a pharmaceutically acceptable derivative thereof. Retroviral infections often attack the subject's central nervous system, and it is a particular advantage of the compounds of this invention that, as experiments have shown, they are able to cross the blood-brain barrier in clinically effective amounts.

It has been found that the compounds according to the invention have a particularly effective activity against retroviral and gram-negative bacterial infections.

As used herein, the term "retroviral infections" refers to any virus that uses a reverse transcriptase during an integral part of its life cycle.

Specific bacteria against which a good activity has been found are the following: Escherichia coli, Salmonella dublin, Salmonella typhosa, Salmonella typhimurium, Shigellaflexneri, Citrobacter friendii, Klebsieila pneumoniae, Vibrio cholera, Vibrio anquillarum, Enterobacter aerogenes, Pasteurella multocida, Haemophilus influerizae, Yersinia enterocolitica, Pasteurella haemolytica, Proteus mirabilis and Proteus vulgaris, the causative organisms of such conditions as traveler's diarrhea, urinary tract infections, bacte- ria (shigellosis), typhoid fever (bovine typhoid fever) and cholera in man, as well as animal diseases such as neonatal enteritis Calves, enteritis in pigs after weaning and colisepticaemia in chickens.

The compounds prepared according to the invention have a particularly good activity against the following viruses: human lymphotropic T-cell viruses (HTLV), in particular 'HTLV-I, HTLV-II and HTLV-III (HIV); Feline leukemia virus, infectious equine anemia virus, goatarthritis virus and other lentiviruses, as well as other human viruses such as hepatitis B virus, Epstein-Barr virus (EBV) and the causative factor of multiple sclerosis (MS). The compounds of the invention have also been found to be effective in the treatment of Kaposi's sarcoma (KS) and the thrombocytopenic purpura (TP). For these latter indications (MS, KS and TP) and goatarthritis virus, the present invention includes the compounds of formula (I) wherein A is thymine, for use in their treatment or prophylaxis, as well as the use of such compounds in the preparation a drug for their treatment or prophylaxis.

The activity of the compounds according to the invention against such a wide range of bacterial and viral infections is clearly a great advantage in medicine, and the new course of action allows the use of these compounds in combination therapy to reduce the possibility of resistance development. However, such combinations are particularly useful because the 3'-azidonucleosides have surprising capacity for potentiation by other therapeutic agents, as described above.

It has been found that 3'-azido nucleosides work synergistically with a wide range of other therapeutic agents, thereby disproportionately increasing the therapeutic potential of both agents. Treatment requires significantly less of each compound, increases the therapeutic ratio and, as a result, reduces the risk of toxicity of each compound

 Compounds of the formula

(I) A

wherein B is a purine or pyrimidine base attached at the 9 or 1 position, or a pharmaceutically acceptable derivative thereof, are suitable for use in combination therapy with at least one secondary therapeutic agent.

In particular, the compound of formula (I) A, or a pharmaceutically acceptable derivative thereof, is suitable for use in the combination therapy as described above when the two active agents are present in a potentiating ratio.

A "potentiating ratio" is that ratio of the compound of formula (I) A, or a pharmaceutically acceptable derivative thereof, to the second therapeutic agent which provides a therapeutic effect greater than the sum of the therapeutic effects of the individual components.

It will be appreciated that although an optimum ratio is usually employed to ensure maximum potentiation, even a minute amount of one agent will be sufficient to potentiate the effect of the others to some extent, and so will any ratio of the two potentiating agents still have the required synergistic effect. However, the greatest synergy is observed when the two agents are in a ratio of 500: 1 to 1: 50.0, preferably 100: 1 to 1: 100, more preferably 20: 1 to 1:20, and most preferably 10: 1 to 1:10 available.

The combinations may suitably be administered together, for example in a unitary pharmaceutical formulation, or separately, for example as a combination of tablets and injections administered at the same time or at different times to achieve the required therapeutic effect.

In antiviral studies, it has been found, for example, that azidonucleosides are potentiated by such various agents as interferons, nucleoside transport inhibitors, glucuronidation inhibitors, renal excretion inhibitors, and even other therapeutic nucleosides which do not necessarily exhibit activity against the same organisms as the compounds of formula (I) A.

Particularly preferred types of interferon are α, β and y, while nucleoside transport inhibitors include such agents as dilazep, dipyridamole 6 - [(4-nitrobenzoyl) thio] -9- (3-D-ribofuranosyl) purine, papaverine, mioflazine, hexobendin , Lidoflazine and their acid addition salts.

Sample leveln is particularly useful in combination with the 3'-azidonucleosides of formula (I) A, as it has both renal excretion inhibiting activity and glucuronidation blocking activity. Examples of other compounds useful in this regard include acetaminophen, aspirin, lorazepam, cimetidine, ranitidine, zomepirac, clofibrate, indomethacin, ketoprofen, naproxen, and other compounds that co-move with glucuronidation or otherwise undergo significant glucuronidation.

3'-Azidonucleosides of Formula I (A) and their pharmaceutically acceptable derivatives are potentiated by other therapeutic nucleoside derivatives as described above, including acyclic nucleosides of the type disclosed, for example, in British Patent 1,528,865, U.S. Pat 4360522, EP-PS 74306, 55239 and 146516 and European Patent Applications 434393 and 434395, and in particular the compounds of the general formula

(A)

CHX CHCH OH, wherein Z is a hydrogen atom or a hydroxy or amino group;

X is (a) an oxygen or sulfur atom or a methylene group and Y is a hydrogen atom or a hydroxymethylene group; or

(b) a methyleneoxy group (-OCH 2) and Y represents a hydroxy group; and pharmaceutically acceptable derivatives thereof.

Examples of the above-mentioned derivatives are salts and esters and include base salts, e.g. Alkali metal (e.g., sodium) or alkaline earth metal salts and pharmaceutically acceptable salts of organic acids such as lactic acid, acetic acid, malic acid or p-toluenesulfonic acid, as well as pharmaceutically acceptable salts of mineral acids such as hydrochloric or sulfuric acid.

Esters of the compounds of formula (A) which may suitably be used in accordance with the present invention include those containing a formyloxy or a C ₁ -T ₆ - (for example a C 1-6 alkanoyloxy (e.g., acetoxy - or propionyloxy-), an optionally substituted aralkanoyloxy (eg a phenyl-Ci ^ alkanoyloxy, such as a phenyl-acetoxy), or an optionally substituted aroyloxy (eg., Benzoyloxy- or Naphthoyloxy-). ) ester group at one or both end positions of the 9-side chain of the compounds of the formula (A) The abovementioned aralkanoyloxy and aroyloxy ester groups may be substituted, for example by one or more halogen atoms (for example chlorine or bromine), or Amino, nitrile or Sulfamidogruppen, wherein the aryl moiety of the group advantageously contains 6 to 10 carbon atoms.

Particularly preferred examples of compounds of the above general formula (A) for use in the present invention include 9 - [(2-hydroxy-1-hydroxymethylethoxy) methyl] guanine, 2-amino-9- (2-hydroxyethoxymethyl) - purine and in particular 9- (2-hydroxyethoxymethyl) guanine (acyclovir). The latter compound has been found to possess a particularly potent potentiating effect, especially when the compound of formula I (A) is 3'-azido-3'-deoxythymidine, as described in the Examples.

In the antibacterial field, it has also been found that a broad spectrum of antibiotics is effective in potentiating the activity of azidonucleosides. These include various agents such as: benzylpyrimidines, e.g. 2,4-diamino-5- (3 ', 4', 5'-trimethoxybenzyl) -pyrimidine (trimethoprim) and analogs thereof, for example those described in British Pat. No. 1,405,246: sulfonamides, e.g. Sulfadimidin; rifampicin; tobramycin; fusidic; chloramphenicol; Clindamycin and erythromycin.

Accordingly, combinations wherein the second agent is at least one of the above-mentioned antiviral or antibacterial agents, or classes of agents, are of great importance.

Other suitable combinations include those wherein the second agent, for example, interleukin II, suramin, phosphonoformate, HPA23, 2 ', 3'-dideoxynucleosides, for example, 2', 3'-dideoxycytidine and 2 ', 3'-dideoxyadenosine, or drugs, such as levamisoi or thy mosin, for increasing lymphocyte counts and / or functions, as appropriate.

It will also be appreciated that the compounds and combinations may also be used in conjunction with other immunomodulatory therapy, such as bone marrow and lymphocyte transplantations. Certain of the retro-viral infections described above, such as AIDS, are usually associated with opportunistic infections. Accordingly, it will be appreciated that, for example, a combination of 3'-azido-3'-deoxythymidine (AZT) and acyclovir would prove particularly useful for the treatment of an AIDS patient with an opportunistic herpes infection, whereas a combination of AZT and 9 - [(2-hydroxy-1-hydroxymethylethoxy) methyl] guanidine would be useful for the treatment of an AIDS patient with opportunistic cytomegalovirus infection.

Very generally preferred pyrimidines of formulas (I) and (I) A for use in the present invention are those of the following formula

R ^{1 is} hydroxy, mercapto, amino, alkylthio, aralkoxy, alkoxy, cyano, alkylamino, wherein the alkyl groups are optionally under

Formation of a heterocycle are connected; R ^{2 is} hydrogen, acyl, alkyl, aroyl or sulfonate; R ^{3 is} hydroxy, mercapto, amino, triazolyl, alkylamino, dialkylamino, where the alkyl groups are optionally substituted to form a

Heterocycle, aralkoxy, alkoxy or alkylthio; R ^{4 is} alkyl, substituted alkyl, halogen, perhalomethyl, hydroxy, alkoxy, cyano, nitro, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl or hydrogen; and pharmaceutically acceptable derivatives thereof.

In the above general formula (I), the dotted lines in the 2 to 6 positions are intended to indicate the presence of single and double bonds in these positions, the relative positions of the single and double bonds being determined by whether the substituents R ¹ and R ^{2 are} groups capable of, for example, keto-enol tautomerism. Preferred pyrimidine nucleoside classes according to the invention are cytidine derivatives, e.g. Compounds of formula (II) wherein R ^{3 is} amino or alkylamino, especially wherein the 3'-azido group is in the erythro configuration ("azido down"); thymidine and uridine derivatives [eg compounds of formula ( I) wherein R ^{3 is} not an amino or alkylamino group] wherein the 3'-azido group is either in the erythro-orertoro configuration ("azido up"); and nucleosides which are unsaturated between the 5C and 6C positions. ,

Very generally preferred purines of the formulas (I) and (I) A for use according to the present invention have the following formula _t

(HA

HO

wherein R ⁶ and R ^{7 may be the} same or different and are selected from amino, hydrogen, hydroxy, mercapto, alkylthio, alkoxy, aralkoxy, cyano or alkylamino; and pharmaceutically acceptable derivatives thereof.

Preferred purine nucleoside classes according to the invention are adenine derivatives, e.g. B. Compounds of formula (II) A wherein R ^{6 is} amino or substituted amino, and guanine derivatives, e.g. B. Compounds of formula (III) wherein R ⁶ , as defined above, is different from amino or substituted amino and R ^{7 is} amino or substituted amino. The above mentioned groups advantageously contain alkyl or aryl groups, as described below. With respect to the above compounds of the formulas (II) and (H) A it should be noted that the above-mentioned alkyl groups advantageously contain 1 to 8 carbon atoms, in particular 1 to 4 carbon atoms, for. For example, methyl or ethyl groups, optionally substituted by one or more suitable substituents, as described below. The above-mentioned aryl groups including the aryl moieties of such groups as aralkoxy are preferably phenyl groups optionally substituted with one or more suitable substituents as described below. The above-mentioned alkenyl and alkynyl groups advantageously contain from 2 to 8, especially 2 to 4, carbon atoms, e.g. Äthenyl or ethynyl, optionally substituted by one or more suitable substituents, as described below.

Suitable substituents on the above-mentioned alkyl, alkenyl, alkynyl and aryl groups are advantageously selected from halogen, hydroxy, C 1-4 -alkoxy, C 1-4 -alkyl, C 12 -aryl, C 1-10 -alkoxy, carboxy, amino and substituted amino, wherein the amino group is monosubstituted or disubstituted by a Ci_4-alkyl radical, and, if it is doubly substituted, the alkyl groups are optionally connected to form a heterocycle.

Other classes of preferred compounds of formula (II) include those in which one or more of R ¹ , R ² , R ³ and R ^{4 are} as defined below, namely R ¹ is hydroxy, mercapto, C 1-4 -alkoxy or amino; R ² is hydrogen, methyl, C _2-2 alkanoyl or benzoyl; R ³ is hydroxy, mercapto, amino, or substituted amino;

R ⁴ is hydrogen if R ^{3 is} amino or substituted amino, and halo, perhalomethyl, C ₂ _ ₃ alkyl, C ₂ -3 alkenyl or substituted ethenyl, if R ^{3 is} not an amino or substituted amino group, and 5 ' Derivatives of such compounds including straight or branched chain alkyl esters optionally substituted with carboxy groups (eg succinate), Ct_ ₆ thioesters, optionally substituted aryl esters, mesylate, glucuronide or mono-, di- or tri-phosphates. Other classes of preferred compounds of formula (II) B include those wherein R ⁶ is amino, Ci ^ alkylamino, mercapto, hydroxy, or C ₁ ^ -alkoxy; and / or R ^{7 is} amino, C 1-4 -alkylamino or hydrogen; and

5'-derivatives of such compounds, including straight or branched chain alkyl esters, optionally substituted with carboxy groups (eg succinate), C-i-g thioesters, optionally substituted aryl esters, mesylate, glucuronide or mono-, di- or triphosphates.

Preferred compounds of formula (I) for use in the treatment or prophylaxis of viral infections are those compounds of formula (II) wherein R ¹ is hydroxy, mercapto, CI_ ₅ alkoxy, amino, C ^^ - aralkoxy or O- Anhydrobinding is what connects the 2- and 5'-positions;

R ^{2 is} hydrogen, methyl or benzoyl;

R ^{3 is} hydroxy, mercapto, amino, substituted amino or C ₁₂ aralkoxy; R ^{4 has} the same meaning as above, provided that A in the formula (I) is not a thymine residue; and

pharmaceutically acceptable derivatives thereof. The azido group is advantageously in the erythroid configuration. Particularly preferred are those compounds as described above which vary only in one of the substituents R ¹ to R ⁴ in the pyrimidine ring from those of a thymine residue (wherein R ¹ and R ^{3 are} hydroxy, R ^{2 is} hydrogen and R ^{4 is} methyl).

Preferred compounds of formula (I) for use in the treatment or prophylaxis of viral infections are those as described above for formula (I), but further include those compounds of formula (I) A wherein B is a thymine Radical, and especially when the mentioned compound is 3'-azido-3'-deoxythymidine.

The compounds of formula (I) which are for use in the treatment or prophylaxis of bacterial infections

preferred are those of formula (H) wherein:.

R ^{1 is} oxygen, sulfur, benzyloxy or methoxy;

R ^{2 is} hydrogen or benzoyl;

R ^{3 has} the same meaning as above;

R ^{4 is} methyl or halo, provided that A in formula (I) is not a thymine residue; and pharmaceutically acceptable derivatives thereof.

The 3'-azido group may be in any configuration, however, the erythro-configuration is particularly preferred, and preferred pharmaceutically acceptable derivatives are the 5'-esters of simple organic acids, preferably from ₁₈ CI_ acids.

Especially preferred are those compounds as described above having only one of the substituents R ¹ to R ⁴ in the pyrimidine ring of those of a thymine residue (wherein R ¹ and R ^{3 are} hydroxy, R ^{2 is} hydrogen and R ^{4 is} methyl ) vary.

Preferred compounds of formula (I) For use in the treatment or prophylaxis of bacterial infections are those as described above for formula (I), but further include those compounds of formula (I) A wherein B is a thymine residue, and especially when said compound is 3'-azido-3'-deoxythymidine.

By a "pharmaceutically acceptable derivative" is meant any pharmaceutically acceptable salt, ester, or salt of such an ester, or any other compound which is capable of administration to the recipient, the parent 3'-azidonucleoside or a therapeutically active metabolite or residue the same (directly or indirectly).

An example of a non-ester compound is the derivative in which the 5'-C and 2-C atoms are linked by an oxygen atom to form an anhydro group.

Preferred esters of the compounds of formula (I) include carboxylic acid esters in which the noncarbonyl portion of the ester group is selected from straight or branched chain alkyl, alkoxyalkyl (e.g., methoxymethyl), aralkyl (e.g., benzyl), aryloxyalkyl (e.g. . phenoxymethyl), aryl (ζ, phenyl, optionally substituted by halogen, C ^^ - alkyl or C ₁ ^ ₄ -alkoxy substituted); Sulfonate esters such as alkyl or aralkylsulfonyl (e.g., methanesulfonyl); and mono-, di- or triphosphate esters. Any reference to any of the above compounds also includes a reference to a pharmaceutically acceptable salt thereof.

With respect to the esters described above, it should be understood that any amount of alkyl present will advantageously contain from 1 to 18 carbon atoms, especially 1 to 4 carbon atoms, unless otherwise specified.

Any aryl moiety present in such esters advantageously comprises a phenyl group.

Examples of pharmaceutically acceptable salts of the compounds of formula (I) and pharmaceutically acceptable derivatives thereof include base salts, e.g. Salts derived from a suitable base, such as alkali metal (e.g.

Sodium), alkaline earth metal (e.g., magnesium) salts, ammonium and NXj (wherein X is Ci_4 alkyl) and mineral acid salts such as the hydrochloride.

Specific examples of pharmaceutically acceptable derivatives of the compound of formula (I) include the monosodium salt and the following 5'-esters: monophosphate; Disodium; diphosphate; Triphosphate, acetate; 3-methyl-butyrate; octanoate; palmitate; 3-chlorobenzoate, benzoate; 4-methyl benzoate; Hydrogensucciniat; pivalate; and mesylate.

Certain compounds prepared according to the invention are new. Accordingly, the invention further provides the compounds of the formula

HO -!

(DB ι

^N 3

wherein C is a purine or a pyrimidine base attached at the 9- or 1-position, respectively, and pharmaceutically acceptable derivatives thereof, other than

(a) compounds of the formula (I) B in which C is an adenine, guanine, uridine, cytidine or thymine base and their 5'-mono- or 5'-triphosphate esters;

(b) the 5'-O-acetate, 5'-0-trityl and 5'-O- (4-methylbenzenesulfonate) derivatives of the compound of formula (I) B wherein C is a uridine base and the 3 ' Azido group in the erythroid configuration;

(c) compounds of formula (I) B wherein C (i) is a 5-bromo-vinyluridine or 5-trifluoromethyluridine radical and the 3'-azido group is in the erythroid configuration; (ii) is a uridine moiety and the 3'-azido group is in the threo configuration; and (iii) is a 5-iodo or 5-fluorouridine moiety and the 3'-azido group is in the erythro or threo configuration; and 5'-O-trityl derivatives of such compounds;

(d) compounds of formula (I) B wherein C (i) is a 5-bromo-vinyluridine or cytidine moiety and the 3'-azido group is in the threo configuration; (ii) is a 5-fluorocytidine moiety and the 3'-azido group is in the erythroid configuration; or (iii) is a 5-methylcytidine residue and the 3'-azido group is in the threo or erythro configuration;

(e) the 5'-O-acetate esters of compounds of the formula (I) B in which C is a 4-chloro-2 (1H) pyrimidinone or 4- (1H-1,2,4-triazole-1 -yl) -2 (1 H) pyrimidinone (optionally substituted at the 5-position by fluorine or methyl) and the 3'-azido group is in the erythro configuration;

(f) the 5'-O - [(4-methoxyphenyl) -diphenylmethyl] derivative of the compound of formula (I) B wherein C is a cytidine moiety and the 3'-azido group is in the erythroid configuration; and

(g) the 5'-O-trityl derivative of the compound of formula (I) B wherein C is an adenine residue and the 3'-azido group is in the threo configuration.

Preferred compounds are those described above in connection with therapy, other than those specifically excluded above.

The compounds of formulas (I) and (I) A and their pharmaceutically acceptable derivatives, also referred to herein as an active ingredient, may be administered for therapy by any suitable route, including oral, rectal, nasal, topical (including buccal and sublingual ), vaginal and parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration. It will be appreciated that the preferred route will vary with the condition and age of the recipient, the nature of the infection, and the active ingredient chosen.

In general, a suitable dose will be in the range of 3.0 to 120 mg per kg body weight of the recipient per day, preferably in the range of 6 to 90 mg kg body weight per day, and more preferably in the range of 15 to 60 mg per kg body weight per day , The desired dose is preferably administered in the form of two, three, four, five, six or more sub-doses at appropriate intervals throughout the day. These sub-doses may be administered in unit dose forms, for example, at a level of 10 to 1500 mg, preferably 20 to 1000 mg, and more preferably 50 to 700 mg, of active ingredient per unit dose form. Trials with 3'-azido-3'-deoxythymidine suggest that a dose to achieve peak plasma active compound concentrations of from about 1 to about 75μ, Μ, preferably from about 2 to 50μ.Μ, more preferably from about 3 to about 30μ .Μ should be administered. This can be accomplished, for example, by the intravenous injection of a 0.1 to 5% solution of the active ingredient, optionally in saline, or orally administered as a bolus at a level of from about 1 to about 100 mg / kg of the active ingredient. Desirable blood levels may be maintained by continuous infusion to provide from about 0.01 to about 5.0 mg / kg / hour or by intermittent infusions at a level of from about 0.4 to about 15 mg / kg of the active ingredient.

The combinations may be administered in a manner similar to that described above to provide the desired therapeutic doses of the active ingredient and the subject additional therapeutic agent. The dosage of the combination will depend on the condition to be treated, the particular active ingredient and the other therapeutic agent concerned, and other clinical factors such as the weight and condition of the patient and the route of administration of the combination. As indicated above, the active ingredient and the other _: therapeutic agent may be administered simultaneously (eg, in a unitary pharmaceutical formulation) or separately (eg, in separate pharmaceutical formulations), and in general, the combinations may be administered topically , oral, rectal or parenteral (e.g., intravenous, subcutaneous or intramuscular) routes. In particular, combinations in which the second agent is a nucleoside transport inhibitor and the subject in question may be administered in a conventional manner. However, for administration by the oral route, a dose of the active ingredient of 1 to 200 mg / kg / day, preferably 5 to 50 mg / kg / day, is generally sufficient. For parenteral administration, a dose of active ingredient of 1 to 100 mg / kg / day, preferably 2 to 30 mg / kg / day, is generally sufficient. The amount of nucleoside transport inhibitor in the combination is independent of the amount of the active ingredient specified above and is sufficient to effectively inhibit nucleoside transport, and is preferably in the range of 0.1 to 100 mg / kg / day, and more preferably in the range of 1 to 20mg / kg / day. The combinations in which the second agent is either a renal excretion inhibitor or a glucuronidation inhibitor or both may be administered to the subject in a conventional manner. However, for administration by the oral route, a dose of active ingredient of 1 to 200 mg / kg / day, preferably 5 to 50 mg / kg / day is usually sufficient. For parenteral administration, a dose of active ingredient of 1 to 100 mg / kg / day, preferably 2 to 30 mg / kg / day, is generally sufficient. The amount of renal excretion / glucuronidation inhibitor in the combination is independent of the amount of active ingredient and is sufficient in the range of 4 to 100 mg / kg / day, preferably 5 to 60 mg / kg / day, more preferably 10 to 40 mg / kg / day.

The combinations in which the second agent is an interferon advantageously contain the active ingredient in amounts of from 5 to 250 mg per kg per day; and the appropriate effective dose range of interferon is 3 x 10 ^e to 10 x 10 ⁶ IU per m ² body surface per day, preferably 4 x 10 ^{6 to 6} x 10 ⁶ IU per m ² per day. The active ingredient and interferon should be present in the ratio of about 5 mg per kg of active ingredient to 3 x 10 ⁶ IU per m ² of interferon to about 250 mg per kg per day of active ingredient to 10 x 10 ⁶ IU per m ² per day Interferon, preferably from about 5 mg per kg per day of active ingredient to 4 x 10 ⁶ IU prom ² per day of interferon to about 100 mg per kg per day of active ingredient to 6 x TO ⁶ IU prom ² per day of interferon ,

Interferon is preferably administered by injection (e.g., s.c., i.m., or i.v.) while the active ingredient is preferably administered orally or by injection, but may be administered in any manner described herein. Interferon and active ingredient may be administered together or separately, and the daily dose of either may preferably be administered in divided doses.

For combinations in which the second agent is another therapeutic nucleoside, a suitable effective oral dose of the active ingredient may be in the range of 2.5 to 50 mg per kg of body weight per day, preferably 5 to 20 mg per kg per day; and the appropriate effective oral dose of the second therapeutic nucleoside is in the range of 5 to 100 mg per kg of body weight per day, preferably 15 to 75 mg per kg per day. It is preferred that the ratio for oral administration of the active ingredient to the second therapeutic nucleoside should be in the range of from about 1: 1 to 1:10, more preferably from about 1: 2 to 1: 8.

The appropriate effective egg. The dose of the active ingredient is generally in the range of about 1.5 to 15 mg per kg per day, and the appropriate effective intravenous dose of the therapeutic nucleoside is generally in the range of about 5 to 30 mg per kg per day Day. It is preferred that the ratio for the i.v. administration of the active ingredient to the second therapeutic nucleoside should be in the range of about 2: 1 to 1:20, more preferably from about 1: 2 to 1:10.

Both components of the combination are preferably administered orally or by injection and they may be given together or separately. The daily dose of each component may be given in divided doses. For combinations in which the second agent is an antibacterial agent, a suitable effective dose of a component is in the range of 2.5 to 50 mg / kg / day, preferably 5 to 10 mg / kg / day, and the appropriate effective dose range of the antibacterial Average is in the range of 2 to 1,000 mg / kg / day, preferably 50 to 500 mg / kg / day. The preferred ratios of active ingredient to antibacterial agent are in the range of 20: 1 to 1: 500, more preferably 2: 1 to 1: 125.

Although combinations with three or more therapeutic agents are not specifically described herein, it will be recognized that such combinations are of great importance. For example, a combination of 3'-azido-3'-deoxythymidine (AZT). With acyclovir and sample levin has the dual advantage of both potentiating the activity of AZT and increasing its availability. Likewise, combinations of the compounds of formula (I) A with two or more compounds of the same series of second therapeutic agent also form part of the invention, for example AZT with sulfadimidine and trimethoprim.

Although it is possible for the active ingredient to be administered alone, it is preferred to present it as a pharmaceutical formulation. The formulations contain at least one active ingredient as defined above together with one or more acceptable carriers therefor and optionally other therapeutic agents. Each carrier must be "compatible" in the sense that it is compatible with the other ingredients of the formulation and is not harmful to the patient Formulations include those for oral, rectal, nasal, topical (including buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous and interdermal) administration The formulations may suitably be presented in unit dosage form and prepared by any method known to those skilled in the art of pharmacy Such methods include the step of combining the active ingredient In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers, or both, and then the product, if necessary, molded.

The formulations suitable for oral administration may be presented as discrete units, such as capsules, cachets or tablets, each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or as a suspension in an aqueous or nonaqueous liquid; or as a liquid oil-in-water emulsion or as a liquid water-in-oil emulsion. The active ingredient may also be presented as a bolus, linctus or paste. '

A tablet may be made by compacting or compressing, optionally with one or more minor ingredients. Compacted tablets may be prepared by compressing the active ingredient in a free-flowing form, such as powder or granules, in a suitable machine, optionally mixed with a binder (eg, povidone, gelatin, hydroxypropylmethylcellulose), lubricant, inert diluent, preservative, dicing agent (e.g., sodium starch glycolate, cross-linked povidone, cross-linked sodium carboxymethylcellulose) and surfactant or dispersant. Compressed tablets can be obtained by compressing a mixture of the powdered compound moistened with an inert liquid diluent in a suitable machine. The tablets may optionally be coated or scored and may be formulated to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethylcellulose in varying ratios to provide the desired release profile.

Suitable formulations for topical administration to the mouth include lozenges containing the active ingredient in a palatable base, usually cane sugar and gum arabic or tragacanth. Pastilles contain the active ingredient in an inert base, such as gelatin and glycerin, or cane sugar and gum arabic; and mouthwash contain the active ingredient in a suitable liquid carrier. Formulations for rectal administration may be presented as a suppository with a suitable base containing, for example, cocoa butter or a salicylate.

Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or sprays containing, in addition to the active ingredient, such carriers as would appear appropriate to one skilled in the art.

Suitable formulations for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may contain suspending and thickening agents. The formulations can be presented in unit dose or multi-dose form in sealed containers, such as ampoules and vials, and stored in a lyophilized state, which merely precedes the addition of the sterile liquid carrier, for example, water for injection requires use. Unprepared injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the type described above. Preferred unit dose formulations are those containing a daily dose or unit, a daily underdose, as indicated above, or a suitable fraction thereof, of an active ingredient.

The compounds of this invention may also be presented for use in the form of veterinary formulations which may be prepared, for example, by methods known to those skilled in the art. Examples of such veterinary formulations include those adapted for

(a) oral administration, for example, a drug (e.g., aqueous or nonaqueous solutions or suspensions); Tablets or boluses; Powders, granules or pellets for a mixture with animal feed; Pastes for application to the tongue;

(b) parenteral administration, for example by subcutaneous, intramuscular or intravenous injection, e.g. as a sterile solution or suspension; or (if appropriate) by intramammary injection, whereby a suspension or solution is introduced into the udder via the teat;

(c) topical application, ζ. B; as a cream, ointment or spray applied to the skin; or

(d) intravaginally, e.g. B. as a pessary, cream or foam.

It will be appreciated that such formulations as described above are also suitable for the presentation of combinations according to the invention, whether uniform or separate formulations, and can be prepared in a similar manner. '

It should be noted that, in addition to the ingredients particularly mentioned above, the formulations may contain other conventional agents relating to the type of formulation in question, for example those suitable for oral administration, such further agents as sweeteners, thickeners and Flavoring may contain. The compounds of formula (I) A and their pharmaceutically acceptable derivatives can be prepared in a conventional manner using procedures known to those skilled in the art, e.g. As described in the following publications: Synthetic Procedures in Nucleic Acid Chemistry (1, [1968], TA Krenitsky et al.), J. Med. Chem. (26, 981 [1983]; Nucleic Acid Chemistry, Improved and New Synthetic Processes, Method and Techniques (Parts 1 and 2, Ed. LD.Townsend, RSTipson [J.Wiley] 1978); JR Horwitz et al. (J. Org. Chem. 29, [JuIi 1964] 2076-78 M.lmazawa et al. (J. Org., Ex., 43 [15] [1978] 3044-3048); KA Watanabe et al (J. Org. Chem., 45, 3274 [1980]); RP Glinski et al (J.Chem.Soc.Chem.Commun., 915 [1970]), which is incorporated herein by reference, or to the use of techniques analogous to those described therein.

The present invention now relates to a process for the preparation of a compound of the formula (I) and of pharmaceutically acceptable derivatives thereof, in which (A) a compound of the formula

(wherein A has the same meaning as above and M represents a precursor group for the 3'-azido group) or a derivative (eg, an ester or a salt) thereof with an agent or under conditions which result in the conversion of the precursor group into the desired azido group , implements; '

a compound of the formula

(IVa)

(wherein R ₃ , R ² , Ri R *, Ra or Rg are the groups R ¹ , R ² , R ³ , R ⁴ , R ⁶ and R ⁷ or precursor groups thereof, provided that at least one of the radicals Ra, Ra, Ra ^and d Ra in the formula (IVa) or at least one of the groups Rg and Ra in the formula (IVb) represent a precursor group), with an agent or under conditions conducive to conversion of the precursor group (s) into the corresponding desired groups lead, implemented; (C) a compound of formula _r

R ³ '

(wherein R ¹ , R ² , R ³ , R ⁴ , R ⁶ and R ^{7 have} the same meaning as above) or a functional equivalent thereof, with a compound capable of introducing the desired ribofuranosyl ring at the 1-position of the compound of Formula (IVa) or the 9-position of the formula (IVb) is used, converts; or

(D) for the preparation of compounds of the formula (II) a purine of the formula (Vb) with a pyrimidine nucleus of the formula

(Vl)

(wherein Py represents a 1-pyrimidinyl group); and subsequently, or concurrently with, one or more of the following transformations, if appropriate:

(i) if a compound of formula (I) is formed, converting the compound into a pharmaceutically acceptable derivative thereof; and

(ii) if a pharmaceutically acceptable derivative of a compound of formula (I) is formed, converting the derivative into the parent compound of formula (I) or into another pharmaceutically acceptable derivative of a compound of formula (I).

It is apparent in the above-described process according to this invention that the choice of precursor compounds in processes (A) to (D) is largely dictated by the particular compound whose preparation is desired, the above-mentioned agents and conditions correspondingly the means and conditions known to those skilled in the art of synthetic nucleoside chemistry. Examples of such conversion methods will be described below for guidance, and it will be understood that they may be modified in a conventional manner depending on the desired compound. In particular, for example, where a conversion is described that would otherwise result in an undesirable reaction of labile groups, such groups may then be conventionally protected with subsequent deprotection after completion of the conversion. ,

Accordingly, for example, with reference to process (A), the group M in the compound of formula (IIIa) or. (1Mb) represent, for example, a halogen (eg chlorine), hydroxy or organosulfonyloxy (eg trifluoromethylsulfonyloxy, methanesulfonyloxy or p-toluenesulfonyloxy) radical.

For the preparation of the compounds of formula (I) in which the 3'-azido group is in the threo configuration, a compound of formula (III a) or (III b) in which the group M is a hydroxy group in the erythro configuration ( in which the 5'-hydroxy group is advantageously protected, eg with a trityl group), for example with triphenylphosphine, carbon tetrabromide and lithium azide. Optionally, M can be an erythro-configuration escaping organosulfonyloxy group which can be converted to an azido group in the threo configuration by treatment, for example with lithium or sodium azide, with the 5'-hydroxy group protected in a similar manner as described above. The removal of the 6'-trityl protecting group can then be effected, e.g. B. by treatment under mild acid conditions or with zinc bromide.

For treating the compounds of formula (I) wherein the 3'-azido group is in the erythro configuration, a compound of formula (IIIa) or (1Mb) in which the group M is a halo group (eg, chloro) in derthreo Configuration (in which the 5'-hydroxy group is advantageously protected, eg, with a trityl group), for example, be treated with lithium or sodium azide. The S'-threo-halogen (eg, chlorine) starting material may be prepared, for example, by reaction of the corresponding 3'-erythro-hydroxy compound with, for example, triphenylphosphine and carbon tetrachloride, or alternatively by treatment with organosulfonyl halide (eg, trifluoromethanesulfonyl chloride) to form a corresponding 3'-erythro-Organosufonyloxyverbindugη which is then halogenated, for example, as described above. On the other hand, a 3'-threo-hydroxy or organosulfonyloxy compound of the formula (IIIa) or (IIIb) can be treated, for example, with triphenylphosphine, carbon tetrabromide and lithium azide to form the corresponding 3'-erythro-azido compound.

Referring to process (B), the following examples are given of various processes by which the precursor groups in formula (IVa) can be converted to the desired R ¹ , R ² , R ³ and R ⁴ groups:

(a) If R ^{1 represents} an alkoxy group (e.g., methoxy or ethoxy), such compounds may be prepared from the corresponding compounds of formula (IVa) in which R 1 represents a 2,5'-O-anhydrobinding, eg by treatment with a suitable nucleophile, eg an alkoxide, suitably in the presence of potassium carbonate;

(b) if R ^{1 is} mercapto, such compounds may be prepared from the corresponding compounds of formula (IVa) in which Rg is an alkoxy group (e.g., ethoxy), for example by treatment with hydrogen sulfide; '

(c) if R ^{2 is} an alkyl group, such compounds can be prepared from the corresponding compounds of formula (IVa) in which R ^{2 is} a hydrogen atom, ζ. B. by treatment with an alkylating agent, for example Ν, Ν-dimethylformamide dimethyl acetal;

(d) if R ^{3 is} mercapto, such compounds may be prepared from the corresponding compounds of formula (IVa) in which R ^{3 is} a suitable leaving group, eg 1,2,4-triazolyl, for example by treatment with an alkali metal (eg, sodium) mercaptan;

(e) if R ^{3 is} an amino group, such compounds may be prepared from the corresponding compounds of formula (IVa) in which R ^{3 is} hydroxy by treatment with an aminating agent, e.g. With hexamethyldisilazane and ammonium sulfate, in a bomb;

(f) if R ^{4 is} halo (e.g., chloro), such compounds may be prepared from the corresponding compounds of general formula (IVa) wherein R 1 is hydrogen by treatment with a halogenating agent, e.g. For example, m-chloroperbenzoic acid.

Similar methods can be used to carry out the conversion of the precursor group in formula (IVb) to the desired R ⁶ and R ⁷ groups, as well as methods / as described, for example, in the above-mentioned references.

The process (C) can be carried out, for example, by treating the appropriate pyrimidine or purine of the formula (Va) or (Vb), or a salt or a protected derivative thereof, with a compound of the formula _r

(wherein Y represents an leaving group, for example, an acetoxy or benzoyloxy or halogen, eg, chloro group, and the 5'-hydroxyl group is optionally protected, for example, by a p-toluenesulfonyloxy group), followed by removal any protective groups.

In process (D), the reaction of the compounds of formulas (Vb) and (VI) is conveniently carried out in the presence of a phosphorylating enzyme and, if desired, effecting the separation of the 3'-azido anomers in a conventional manner.

If a compound of formula (I) is formed, such a compound may be converted into a pharmaceutically acceptable phosphate or other ester by reacting the compound of formula (I) with a corresponding phosphorylating agent,

z. B. POCl ₃ or a suitable esterifying agent, for. As an acid halide or anhydride are converted. The compounds of formula (I), including their esters, may be converted into pharmaceutically acceptable salts thereof in a conventional manner, e.g. By treatment with a suitable base.

Where a derivative of a compound of formula (I) is formed, such a compound may be converted into the parent compound, e.g.

by hydrolysis.

The following examples are merely illustrative of the invention and are not intended to limit it in any way. The term "active ingredient" as used in the examples means a compound of formula (I) or (I) A, or a pharmaceutically acceptable derivative thereof.

embodiments

Example 1 Tablet Formulations

The following formulations A and B were prepared by wet granulation of the ingredients with a solution of povidone followed by addition of magnesium stearate and densification.

mg / tablet mg / tablet Formulation A (a) Active ingredient 250 250 (b) lactose B.P. 210 26 (c) povidone B.P. 15 9 (d) Sodium starch glycolate 20 12 (e) magnesium stearate 5 3 500 300 Formulation B mg / tablet mg / tablet (a) Active ingredient 250 250 (b) lactose 150 - (c) Avicel PH101 60 26 (d) Povidone B.P. 15 9 (e) Sodium starch glycolate 20 12 (f) magnesium stearate 5 3 500 300 Formulation C mg / tablet Active stock part 100 lactose 200 Strength 50 povidone CJL magnesium stearate 4 359

, The following formulations, D and E, were prepared by direct compression of the blended ingredients. The lactose used in formulation E is of the direct compression type (Dairy Crest "Zeparox").

Formulation D

Active ingredient Pregelatinized starch NF15 mg / capsule 250 150 400 Formulation E Active Ingredient Lactose Avicel mg / capsule 250 150 100 500

Formulation F (Controlled Release Formulation) The formulation is prepared by wet granulation of the ingredients (below) with a solution of povidone followed by addition of magnesium stearate and densification.

mg / tablet

(a) Active ingredient 500

(b) Hydroxypropylmethylcellulose 112 (Methocel K4M Premium)

(c) lactose B.P. 53

(d) povidone B.P.C. 28

(e) magnesium stearate 7

700

Example 2 Capsule Formulations

Formulation A

A capsule formulation is prepared by mixing the ingredients of Formulation D in Example 1 above and filling into a two-part hard gelatin capsule. Formulation B (below) is obtained in a similar manner.

Formulation B

mg / capsule (a) Active ingredient 250 (b) lactose B.P. 143 (c) Sodium starch glycolate 25 (d) magnesium stearate 2 420 Formulation C mg / capsule (a) Active ingredient 250 (b) Macrogol 4000BP 350 600

The capsules were prepared by melting the Macrogol 4000 BP, dispersing the active ingredient in the melt and filling the melt into a two-part hard gelatin capsule.

Formulation D

mg / capsule

Active ingredient · 250

Lecithin 100

Peanut oil 222

450

The capsules were prepared by dispersing the active ingredient in the lecithin and peanut oil and filling the dispersion into soft gelatin elastic capsules.

Formulation E (Controlled Release Capsule) The following controlled release capsule formulation was prepared by extruding ingredients (a), (b) and (c) using an extruder, followed by spheronization of the extrudate and drying. The dried pellets were then coated with a release controlling membrane (d) and filled into a two-piece hard gelatin capsule.

mg / capsule

(a) Active ingredient 250

(b) Microcrystalline cellulose 125

(c) Lactose BP125 125

(d) ethyl cellulose 13

513

Example 3 Injectable formulation Formulation A

Active ingredient 0.200 g

Hydrochloric acid solution, 0.1 molar q.s. to pH 4.0-7.0 sodium hydroxide solution, 0.1 molar q.s. to pH 4.0-7.0 sterile water q.s. to 10 ml

The active ingredient was dissolved in the bulk of the water (35 ^° C. to 40 ^° C.), and the pH was adjusted to between 4.0 and 7.0 with the hydrochloric acid or the sodium hydroxide, as the case may be. The batch was then made up to volume with the water and filled through a sterile micropore filter into a sterile amber 10 ml glass vial (Type 1) and sealed with sterile closures.

Formulation B

Active ingredient 0.125 g

Sterile, pyrogen-free

pH7 phosphate buffer · q.s. to 25 ml

Example 4 Intramuscular injection

Weight (g)

Active ingredient 0.20

Benzyl alcohol 0.10

Glycofurol 75 '1.45

Water for injection q.s. to 3.00 ml

The active ingredient is dissolved in the glycofurol. The benzyl alcohol is then added and dissolved and water made up to 3 ml. The mixture is then filtered through a sterile micropore filter and placed in a sterile amber 3 ml glass vial (Type 1) and sealed.

Example 5 Syrup

Formulation A

Active ingredient

Sorbitol solution

glycerin

sodium benzoate

Flavor, peach

Purified water

The active ingredient is dissolved in a mixture of the glycerol and the bulk of the purified water. An aqueous solution of the sodium benzoate is then added to the solution, followed by the addition of the sorbitol solution and finally the flavor. The volume is filled up with purified water and mixed well. When the active ingredient is poorly soluble, the following formulation (B) is used.

Weight (g) 0.2500 1.5000 2.0000 0.0050 17.42.3169 0,0125ml q.s. to 5,0000 ml

Formulation B

Weight (g)

Active ingredient. 0,250

Sorbitol solution 1,500

Glycerol 0.005

Dispersible cellulose 0.005 '.

Sodium benzoate 0.010 ml

flavoring

Purified water to 5,000 ml

The sorbitol solution, the glycerine and part of the purified water are mixed. The sodium benzoate is dissolved in purified water and the solution added to the bulk. The dispersible cellulose is added and dispersed with the flavor. The active ingredient is added and dispersed. The volume is filled up with purified water.

Example 6 Suppository

mg / suppository

Active Ingredient (63mm) * 250

Hard fat, BP (Witepsol H 15-Dynamit Nobel). 1770

2 020

The active ingredient was used as a powder wherein at least 90% of the particles had a diameter of 63μ.η or less.

One fifth of the Witepsol H 15 amount is melted in a pan with steam jacket at a maximum of 45 ⁰ C. The active ingredient is sieved through a 200 μm sieve and added to the molten base with mixing using a Silverson provided with a cutting head until a smooth dispersion is obtained The mixture was held at 45 ° C , the remaining Witepsol H15 was added to the suspension and stirred to ensure a homogeneous mixture.The entire suspension was sieved through a 250 »m stainless steel screen and allowed to cool to 40 ^° C. with constant stirring at a temperature of 38 ^° C. to 40 ^° C. ⁰ C, 2.02 g of the mixture was poured into suitable 2 ml plastic molds and the suppositories were allowed to cool to room temperature.

Example 7 mg / pessary pessaries 250 380 Active ingredient (63 / xm) 363 Anhydrous dextrose 7 potato starch 1000 magnesium stearate

The above ingredients were mixed directly and pessaries prepared by direct compression of the resulting mixture. The following Examples 8-10 illustrate the preparation of formulations containing a compound of Formula (I) A (Active Ingredient) and another therapeutic Nuceloside as indicated. I ι

Example 8 Injection

Formulation A '

Acyclovir Active Ingredient imolareNaOH SterileWater

Formulation B

2-Amino-9- (2-hydroxyethoxymethyl) purine Active Ingredient Imolar NaOH Sterile Water

In the above formulations, the therapeutic nucleoside was added to 1 molar NaOH solution and stirred until dissolved. The active ingredient is added and dissolved. The volume is made up to 10 ml with sterile water. It is filtered through a sterile filter and filled into sterile vials. Trockenfrierung.

Weight (mg) 400 200 Upon need on 10ml Weight (mg) 400 200 Upon need on 10ml

Example 9 Weight (mg) tablets 500 Formulation A 125 14 Acyclovir 25 Active ingredient 6 povidone 670 sodium starch magnesium stearate Weight (mg) 125 Formulation B 125 Active ingredient 8th 2-amino-9- (2-hydroxyäthoxy- 12 methyl) purine 3 povidone 273 sodium starch magnesium stearate

In the above formulations A and B, the therapeutic nucleoside and the active ingredient were mixed with the sodium starch glycolate and granulated with a povidone solution, after drying, the granules were mixed with magnesium stearate and compacted.

Example 10 capsules

Formulation A

Weight (mg)

Acyclovir 250

Active ingredient 62.5

Lactose. 170.5

Sodium starch glycolate 15

Magnesium stearate 2

500

Mix the ingredients and pour into hard gelatin capsules. Formulation B

Weight (mg) Active ingredient 125 2-amino-9- (2-hydroxyäthoxy- methyl) purine 125 lactose 133 sodium starch 15 magnesium stearate 2 400

Mix the ingredients and pour into hard gelatin capsules.

Example 11 illustrates a formulation containing interferon and a compound of formula (I) A (active ingredient).

Example 11 Injection

Interferon · 3Mega units

Active ingredient 200 mg

Sterile buffer, pH 7 to 50 ml

Dissolve the interferon and the active ingredient in the sterile water. It is filtered through a sterile filter and filled into sterile vials.

The following Examples 12-15 describe the preparation of formulations containing a compound of Formula (I) A (the active ingredient), for example, 3'-azido-3'-deoxythymidine, in combination with a nucleoside transport inhibitor, e.g. B.

Dipyridamole.

Example 12 total weight Weight (mg) tablet 300 200 Nucleosidtransportinhibitor 105 Active ingredient 50 lactose 20 Strength 10 polyvinylpyrrolidinone 685 magnesium stearate

The active compounds are mixed with the lactose and the starch and granulated wet with a solution of the polyvinylpyrrolidinone. The granules are dried, sieved, mixed with magnesium stearate and then compressed into tablet form.

Example 13 total weight Weight (mg) capsule 300 100 Nucleosidtransportinhibitor 100 Active ingredient 10 lactose 10 sodium starch 3 polyvinylpyrrolidinone 523 magnesium stearate

The active compounds are mixed with the lactose and the sodium starch glycolate and wet granulated with a solution of the polyvinylpyrrolidinone. The granules are dried, sieved, mixed with the magnesium stearate and filled into hard gelatin capsules.

Example 14 cream

Weight (g)

Nucleoside transport inhibitor 7.50

Active ingredient '5.00

Glycerin 2.00

Cetostearyl alcohol 6,75

Sodium lauryl sulfate 0.75

White soft paraffin 12.50

Liquid paraffin 5.00

Chlorocresol 0.10

Purified water to 100.00

The active compounds are dissolved in a mixture of purified water and glycerol and heated to 70 ° C. The remaining components are heated together to 70 ⁰ C. The two parts are combined and emulsified. The mixture is cooled and bottled. ^! !

Ί ι

Example 15 Intravenous injections

(1) Active ingredient nucleoside transport inhibitor glycerol

Sodium hydroxide solution Water for injections

The glycerin is added to a portion of the water for injections. The two active compounds are added and the pH is adjusted to 7.0-7.5 with sodium hydroxide solution. The solution is made up to volume with additional water for injections. Under aseptic conditions, the solution is sterilized by filtration, filled into sterile ampoules and closed the ampoules.

(2) Active ingredient nucleoside transport inhibitor mannitol

Sodium hydroxide solution Water for injections

Quantity (mg) 200 300 200 q.s.auf pH 7.0-7.5 on 10ml

Quantity (mg) 100 150 125 q.s.auf pH 8.0-9.0 on 2.5 ml

The active compounds and mannitol are dissolved in a portion of the water for injections. The pH is adjusted to 8.0 to 9.0 with the sodium hydroxide solution and made up to volume with additional water for injections. The solution is sterilized by filtration under aseptic conditions, filled into sterile vials and the water removed by lyophilization. The vials are shifted under a nitrogen atmosphere and fitted with a sterile cap and metal car.

The following Examples 16-18 describe the preparation of formulations using a compound of Formula I (A) (the active ingredient), for example, 3'-azido-3'-deoxythymidine, in conjunction with a glucuronidation / renal excretion inhibitor, for example, sample , contain.

Example 16 Weight (mg) - 200 tablet 100 105 50 Active ingredient 20 Glucuronidation / renal excretion inhibitor 10 lactose 485 Strength polyvinylpyrrolidinone total weight Magnesium stearate.

The active compounds are mixed with the lactose and the starch and wet granulated with a solution of the polyvinylpyrrolidinone. The granules are dried, sieved and mixed with magnesium stearate and then compressed into tablet form.

BeispieM7 total weight Weight (mg) capsule 100 Active ingredient 100 Glucuronidation / Nierenexkretion- 100 inhibitor 10 lactose 10 sodium starch 3 polyvinylpyrrolidinone 323 magnesium stearate

The active compounds are mixed with the lactose and the sodium starch glycolide and granulated wet with a solution of the polyvinylpyrrolidinone. The granules are dried, sieved and mixed with the magnesium stearate and filled into hard gelatin capsules.

Example 18 Intravenous injections

Quantity (mg)

(1) Active ingredient 200 glucuronidation / renal excretory

tion inhibitor 300

Glycerol 200

Sodium hydroxide solution q, s. to pH 7.2-7.5

Water for injections to 10 ml

The glycerin is added to a portion of the water for injections. The two active compounds are added and the pH adjusted to 7.0-7.5 with sodium hydroxide solution. The solution is made up to volume with additional water for injections. The solution is sterilized by filtration under aseptic conditions, filled into sterile ampoules and the vials are closed.

q.s. on Quantity (mg) Active ingredient on 100 Glucuronidation / Nierenexkre- tion inhibitor 150 mannitol 125 Sodium hydroxide solution pH 8.0-9.0 Water for injections 2.5 ml

The active compounds and mannitol are dissolved in a portion of the water for injections. The pH is adjusted to 8.0 to 9.0 with the sodium hydroxide solution and made up to volume with additional water for injections. The solution is sterilized by filtration under aseptic conditions, filled into sterile vials and the water removed by lyophilization. The vials are sealed under a nitrogen atmosphere and fitted with a sterile cap and metal collar

Veterinary formulations (Examples 19 to 22) Example 19 Tablet for use in small animals

Per tablet

Active ingredient "120.0 mg

Corn starch 20.0 mg -

Microcrystalline cellulose 100.0 mg

Magnesium stearate 1.5 mg

The active ingredient, the microcrystalline cellulose and the corn starch are mixed together. Sufficient starch solution is added with continued mixing to produce a wet mass, which is then passed through a sieve to form granules. The magnesium stearate is sprinkled through a sieve. The tablets are made by direct compression of the granules.

Example 20 Weight (g) Granules for a drug in the feed 6.0 1.0 Active ingredient 93.0 povidone lactose Aqueous alcohol mixture q. st

The active ingredient is mixed with the lactose. For this, the aqueous alcohol containing the dissolved povidone is added. Sufficient aqueous alcohol is added to form a wet mass which is passed through a sieve to produce a granulate which is subsequently dried.

Example 21

Oral paste ,

Weight (g)

Active ingredient 24

Xanthan gum. 0.5

Methyl hydroxybenzoate 0.1

Polysorbate80 0.1

Purified water to 100.0 ml

Polysorbate 80 and methyl hydroxybenzoate are dissolved in the bulk of the water. The xanthan gum is added and dispersed and allowed to swell. The active ingredient is added, dispersed and diluted to volume.

Example 22 Ointment

Weight (g)

Active ingredient 12

White soft paraffin 88.0

The white soft paraffin is melted at 60 ^° C. The active ingredient is added and dispersed, allowed to cool, and filled into collapsible metal tubes.

Example 23 3'-Azido-3 ', 5'-dideoxy-5' - [(N _, N-dimethylthiocarbamoyl) thid] thymidine

(a) The 5'-hydroxyl group of 3'-azido-3'-deoxythymidine (3.0 g, 11.2 mmol) was converted to a solution of 3'-azido-3'-deoxythymidine by addition of methanesulfonyl chloride (2.7 ml) in mesylated dry pyridine (20 ml). The reaction was allowed to proceed at 5 ° C for 1 hour and then poured into ice-water. The precipitate was collected by filtration. The desired product was obtained by reacting the 3'-azido-3'-deoxy-5'-mesylthymidine obtained in the first step with potassium carbonate (0.78 g, 5.6 mmol) in DMF (75 ml). The reaction mixture was heated in an 80 ^° C oil bath for 6 hours and then poured into ice-water. The product was extracted from the water with ethyl acetate. The solvent was removed in vacuo and the resulting oil on silica gel rapidly chromatographed by elution with CHCl ₃ : MeOH (9: 1 v / v). The product was obtained in low yield. Melting point = 184 ° C to 186 ° C. ,

b) The sodium salt of dimethyldithiocarbamic acid. Dihydrate (0.642 g, 3.58 mmol) and 3.58 mL of a solution of 1 N-tetrabutylammonium hydroxide in MeOH were added to 25 mL of DMF. The solution was boiled to remove water and MeOH. After cooling, 2,5'-O-anhydro-3'-azido-3'-deoxythymidine (0.85 g, 3.4 mmol) dissolved in 15 ml of DMF was added. The reaction mixture was heated at 55 ^° C. overnight in an oil bath. The reaction mixture was poured onto ice-water and a precipitate removed by filtration. The product was extracted from the filtrate with ethyl acetate. The ethyl acetate was removed in vacuo and the resulting oil was purified by flash chromatography on silica gel eluting with CH 2 Cl 2 -TMeOH (95: 5 v / v). It was necessary to chromatograph a second time on silica gel. The second elution was with CHCl ₃ : MeOH (98: 2 v / v). The final purification was carried out by reverse phase chromatography on Ci ₈ , eluted with water-methanol (3: 7). The yield was 2.5%.

β. ·

Example 24 S'-azido-S'-deoxy-S'-O-acetyl-thiothymidine

S'-azido-S'-deoxy-O'-O-acetyl ^ -d ^ atriazo-D-thymidine (Lin et al., J. Med. Chem. 26, 1691 [1983]) (1.41 g, 3, 9 mmol) was dissolved in 100 ml acetone and 30 ml water and then with 0.39 g NaSH. H ₂ O (Sung, J. Chem. Soc. Chem. Comm., 522, [1982]). The mixture was stirred for 30 minutes, the volume reduced to half and extracted with 200 ml of CHCl ₃ . The CHCl ₃ was washed with 100 ml of water and dried over sodium sulfate. The solvent was removed in vacuo and the resulting oil placed on a silica gel pad (6.5 x 3 cm), followed by elution with 750 ml of CHCl ₃ . The solvent was removed in vacuo to give a yellow oil which, upon crystallization from i-PrOH, gave a yield of 0.64 g (1.9 mmol, 48.7%). Melting point = 75 ° C to 78 ° C.

Example 25 3'-Azido-3'-deoxy-4-thiothymidine

3'-Azido-3'-deoxy-5'-O-acetyl-4-thiothimidine (0.25 g, 0.76 mmol, Example 21) was dissolved in a mixture of 5 mL dioxane and 5 mL conc. NH ₄ OH and 18 hours stirred for a long time. The solvent was removed in vacuo and the residue was applied to a silica gel column, followed by elution with CHCl ₃ / EtOAc (3: 1 v / v). The appropriate fractions were combined and the solvent removed in vacuo. A yellow oil was obtained which dissolved in Et ₂ O to yield crystals: 0.16 g (0.56 mmol, 74%). Melting point = 116 ° C to 118 ⁰ C.

Example 26 3-N-Methyl-3'-azido-3'-deoxythymidine

3'-azido-3'-deoxythymidine (0.5 g, 1.9 mmol) and N, -dimethylformamide dimethyl acetal (Zemlicka, Coll. Czech Chem. Comm. 35, 3572 [1972]) (0.9 ml, 7.5 mmol) were refluxed in 20 ml of CHCl _{3 for} 48 hours. The solvent was removed in vacuo and the material placed on a silica column. Elution with EtOAcZCHCl ₃ (1: 1 v / v) yielded a pure material in the form of a viscous oil: 0.26 g (0.9 mmol, 47%).

Example 27 3'-Azido-3'-deoxy-2-thiothymidine

The synthesis of 3'-azido-2-thiothymidine was carried out by a five-step reaction sequence starting from 2,3'-O-anhydro-5'-tritylmidine (JJ Fox, J. Org. Chem., 28, 936 [1963]).

2,3'-O-Anhydro-5'-tritylthymidine (10.5 g, 22.4 mmol) was added to a solution of sodium (0.52 g, 22.4 mmol) in dry ethanol (1.2 liters) and the reaction mixture heated at reflux for 6 hours. The reaction mixture was cooled and neutralized with 1N HCl. The solvent was removed in vacuo and the resulting oil purified by flash chromatography on silica gel eluting with CHCl ₃ : MeOH (96: 4 v / v). There was obtained a 30% yield of 1- (2'-deoxy-5'-trityl-D-lyxofuranosyl) -2-ethoxythymine. The 2-ethoxythymidine derivative (3.5 g, 16.8 mmol) was dissolved in 35 ml of DMF containing 2.2 ml of triethylamine. The cold solution was saturated with H ₂ S. The reaction mixture was transferred in a steel bomb and heated to 95 ⁰ C. After 27 hours, thin layer chromatography (TLC) revealed no starting material present. The reaction mixture was purged with N _{2 for} several hours and poured into ice-water. The product was collected by filtration and purified by flash chromatography on silica gel eluting with CHCl ₃ : MeOH (97: 3 v / v). A 37% yield of 1- (2'-deoxy-5'-trityl- / 3-D-lyxofuranosyl) -2-thiothymine was obtained. The UV maxima at pH 1 of 277 nm and at pH 11 of 241 nm indicated the formation of a 2-thiothymidine. The 3'-hydroxyl group of the thiothymidine derivative was mesolyzed as follows: Methanesulfonyl chloride (665 ml, 3.5 equivalents) was added in 4 portions over 6 hours to a solution of 1- (2'-deoxy-5'-trityl - / 3-D-lyxofuranosyi) -2-thiothymidine (1.25g) in dry pyridine (15ml) at 5 ° C. The reaction mixture was kept at 5 ° C overnight. The reaction mixture was poured onto ice-water and the product collected by filtration. Purification was performed by flash chromatography on silica gel eluting with ethyl acetate: hexane (1: 1 v / v). The yield was 50%.

Lithium azide (0.3 g, 6 mmol) was dissolved in 20 ml dry DMF and 1- (2'-deoxy-3'-mesyl-5'-trityl- / 3-D-lyxofuranosyl) -2-thiothymine (0.72 g, 1 , 2 mmol) was added. The DMF solution was heated at 85 ° C for 2.5 hours. The reaction mixture was poured into ice-water and the product collected by filtration. Purification was performed by flash chromatography on silica gel eluting with CHCl ₃ : MeOH (98: 2 v / v). The yield was 78%. An IR band at 2100 cm ^-1 indicated the presence of an alkyl azide, UV confirmed the presence of a 2-thiothymidine, and the final product was prepared by deblocking the 5'-hydroxyl group of 3'-azido-3'-deoxy-2-thio. 5'-tritylthymidine (0.1 g) in 80% acetic acid (5 ml) on a steam bath over a period of 45 minutes. "3'-Azido-3'-deoxy-2-thiothymidine (0.021 g) by chromatography on silica gel eluting with CHCl ₃ : MeOH (96: 4 v / v) in 37% yield.

Example 28 3'-Azido-3'-deoxy-2-ethoxythymidine

3'-Azido-3'-deoxy-2-ethoxythymidine was prepared by refluxing 3'-azido-3'-deoxy-5'-mesylthymidine (2.6g, 7.5 mmol) in dry ethanol (25 ml) with two Equivalent potassium carbonate (1.08 g, 7.5 mmol) was produced over a period of 5 hours. The solution was neutralized and concentrated in vacuo to an oil. The oil was purified by flash chromatography on silica gel eluting with ethyl acetate: methanol. The desired product was isolated in 39% yield; Melting point = 98 ° C to 100 ⁰ C.

Example 29 3'-Azido-3'-deoxy-2-methoxythymidine

3'-Azido-3'-deoxy-2-methoxythymidine was prepared from 3'-azido-3'-deoxy-5'-mesylthymidiri (1.6g, 4.6moles) following the procedure of Example 25. The yield was 42%; Melting point = 47 ⁰ C to 51 ⁰ C.

Example 30 3'-azido-2 ', 3'-dideoxy-5-methylisocytidine

2,5'-O-Anhydro-3'-azido-3'-deoxythymidine (0.35 g, 1.4 mmol) was placed in a bomb in 15 ml of MeOH pre-saturated with ammonia and heated at 77 ° C (oil bath) for 48 hours (Skaric and Matulic-Adamic, HeIv. Chim. Acta, 63, 2179 [1980]). TLC (6: 1 v / v CHCl ₃ / MeOH) indicated that the reaction was incomplete. The solvent was removed in vacuo and the resulting oil applied to a silica gel column followed by elution with CHCl ₃ / MeOH (6: 1 v / v). The appropriate fractions were combined to give a yield of 0.14 g (0.53 mmol, 38%) of the title compound; Melting point = 107 ° C to 108 ⁰ C.

Step Example! 31 3'-azido-5-chloro-2 ', 3'-dideoxyuridine

3'-azido-2 ', 3'-dideoxyuridine (0.25 g, 1 mmol) was dissolved in 2 ml of dry dimethylacetamide (DMAC), cooled to 0 ⁰ C, and 2 ml of a 0,5molaren HCI in DMAC was added. M-chloroperbenzoic acid (0.277 g, 1.6 mmol) was added in two portions over 10 minutes, after which the mixture was allowed to cool to room temperature. After 2 hours, 4 ml of H ₂ O were added and the solution filtered. The aqueous DMAC solution was extracted with Et ₂ O (3x3ml) and the Et ₂ O evaporated in vacuo to give an oil which was applied to a silica gel column. Elution with CHCl ₃ : MeOH (15: 1 v / v), combination of the appropriate fractions, and evaporation in vacuo afforded an oil which was recrystallized from Et ₂ O. 58.5 mg (0.2 mmol, 20%) were obtained. Melting point = 169 ° C to 17O ⁰ C. UV (nm): at pH 1 Amax = 276 (ε = 7400), amine = 239 (ε = 500);

at pH 13Amax = 274 (ε = 6400), amine = 249 (ε3800).

H ¹ NMR (DMSO-d ₆ ): δ 8.29 (s, 1H, H 6), 6.04 (t, 1H, H 1 ', J = 5.5Hz), 5.49-5, 29 (m, 1H, 5'-OH), 4.44 ^ 1.20 (m, 1H, H3 '), 3.88-3.71 (m, 1H, H4'), 3.71 -3.53 (m, 2H, H5 '), 2.63-2.31 (m, 2H, H2'); Analysis for C ₉ H ₁₀ N ₅ O ₄ Cl:

Calculated: C 37.58%, H 3.50%, N 24.35%, Cl 12.32%; Found: C37.67%, H3.54%, N24.39%, Cl 12.40%.

Example 32 3'-azido-5-bromo-2 ', 3'-dideoxuridine

3'-azido-5-bromo-2 ', 3'-dideoxyuridine was prepared from the known 3'-azido-2', 3'-dideoxyuridine (TA Krenitsky et al., J. Med. Chem., 26, 891 1983]) (0.827 g, 3.3 mmol) by acetylating the 5'-hydroxyl group with acetic anhydride (15 ml) followed by bromination 5-position by addition of acetic acid (0.5 ml) and bromine (0.566 g). The red-brown solution was stirred for 2 hours at room temperature. The reaction mixture was evaporated in vacuo to an oil and triturated with ethyl ether. The oil was dissolved in methanol / ammonia to remove the acetyl group. The desired product was isolated by chromatography on silica gel eluting with CHCl ₃ : MeOH (95: 5 v / v). The yield was 32%. Melting point = 148 ° C to 149 ° C.

Example 33 3'-azido-2 ', 3'-dideoxy-5-iodouridine

3'-azido-2 ', 3'-dideoxy-5-iodouridine was prepared from 2', 3'-dideoxy-5-iodouridine (10 g, 28 mmol) by a four-step reaction sequence described in the literature (TA Krenitsky et al. J. Med. Chem., 26, 891 [1983]). Melting point = 126 ⁰ C to 130 ⁰ C.

Step Example! 34 S'-azido-Z '^' - dideoxy-S-trifluoromethyluridine

3'-azido-2 ', 3'-dideoxy-5-trifluoromethyluridine was prepared by the following four-step reaction sequence. The 5'-hydroxyl group of 2 ', 3'-dideoxy-5-trifluoromethyluridine (5.0g, 16.9mmol) was added to the starting material in a suspension of dichloromethane (1mL by addition of triphenylmethylchloride, 5.65g, 20.3mmol) , 4 liters of ml), pyridine (70 ml) and 3A molecular sieves (55 g) tritylated. The reaction mixture was stirred at room temperature for 4 days. After filtration and evaporation in vacuo, the oily product was chromatographed on silica gel and chromatographed with CH ₂ Cl ₂ : MeOH (95: 5 v / v). The product fractions were combined and the solvent removed in vacuo. The resulting oil was triturated with water and the resulting solid collected by filtration. The 3'-hydroxyl was chlorinated by dissolving the 5'-protected uridine (3.0g) in dimethylacetamide (30ml) containing triphenylphosphine (3.27g) and carbon tetrachloride (51ml) was added. The reaction mixture was stirred at room temperature overnight. 1 ml of methanol was added. The reaction mixture was evaporated to an oil and chromatographed on silica gel, eluting with CH ₂ Cl ₂ : EtOAc (9; 1 v / v). The desired product was then collected as an oil. The oil, 1- (3'-chloro-2'-deoxy-5'-trityl-threo / 3-D-ribofuranosyl) -5-trifluoromethyluracil, was prepared by dissolving in nitromethane (80 ml) and adding zinc bromide (4, 45g), dissolved in nitromethane (80ml) according to the method of V. Kohli et al., Tetrahedron Letters, 21, page 2683, 1980, deblocked. The reaction mixture was stirred at room temperature overnight. The next day, additional zinc bromide (3.0 g) was added and the reaction allowed to stand overnight. A final addition of zinc bromide (3.7 g) with gentle heating brought the reaction to an endpoint. The reaction mixture was poured into 1 molar ammonium acetate. The product was extracted with dichloromethane. The dichloromethane was removed in vacuo and the resulting oil chromatographed on silica gel, eluting with CH ₂ Cl ₂ : MeOH (95: 5 v / v). The final product was prepared by treating i-P'-chloro '- deoxy-β-D-ribofuranosyl J-S-trifluoromethyluracil (0.48 g, 1.53 mmol) with lithium azide (0.19 g, 3.8 mmol) in dimethylacetamide (4 , 8ml). The reaction mixture was heated to 90 ° C for 4 hours. The reaction mixture was concentrated to an oil and chromatographed on silica gel, eluting with CHCl ₃ : MeOH (95: 5 v / v). It was necessary to chromatograph a second time. Elution on silica gel with CH ₂ Cl ₂ : MeOH (97: 3 v / v) gave a substantially pure product. Recrystallization from toluene gave the pure product in 10% yield.

Example 35 3'-azido-2 ', 3'-dideoxycytidine

3'-Azido-2 ', 3'-dideoxycytidine was prepared from 3'-azido-2', 3'-dideoxyuridine (2.2g, 7.9 mmol) as HCl salt by the method of TAKrenitsky et al., J. Med Chem., 26, 891 (1983). The yield was 40%. Melting point = 174.5 to 176.5 ⁰ C ⁰ C.

Example 36 3'-azido-2 ', 3'-dideoxy-5-methylcytidine

3'-azido-2 ', 3'-dideoxy-5-methylcytidine was prepared by the method of Example 35 from 3'-azido-3'-deoxythymidine (0.8g, 3, OmMoI). The yield was 19%.

Example 37 Threo-3'-azido-2 ', 3'-dideoxycytidine

, The synthesis of threo-3'-azido-2 ', 3'-dideoxycytidine was carried out from 2'-deoxyuridine in four steps. The 5'-hydroxyl group of 2'-deoxyuridine was tritylated according to the procedure described in Synthetic Procedures in Nucleic Acid Chemistry, 1.321 (1968).

Threo-3'-azido-2 ', 3'-dideoxy-5'-trityluridine was prepared by reacting 2'-deoxy-5'-trityluridine (5.0 g, 10.6 mmol) with triphenylphosphine (3.07 g, 11%). 7 mmol, 1.1 equiv) and carbon tetrabromide (3.88 g, 11.7 mmol, 1.1 equiv) and lithium azide (5.21 g, 106 mmol, 10 equiv) in DMF (80 mL). The carbon tetrabromide was finally added. The reaction mixture was allowed to cool to room temperature overnight. Methanol (5 ml) was added. The solution was concentrated in vacuo to an oil and flash chromatographed on silica gel, eluting with ethyl acetate. The blocking of the 5'-hydroxy position was carried out by heating in 80% acetic acid on a steam bath for a period of 20 minutes. Upon cooling, the tritylcarbinol precipitated and was filtered off. The filtrate was evaporated to dryness and slurried in ethyl ether. The product, threo-3'-azido-2 ', 3'-dideoxyuridine, was used without further purification. The final product, threo-3'-azido-2 ', 3'-dideoxycytidine, as HCl salt, was prepared from the uridine analogues by the exact same procedure used to prepare the erythro isomer (TA Krenitsky et al. , J. Med. Chem., 26, 891 [1983]). The yield was 0.021 g; 7%.

Example 38 9- (3'-azido-2 ', 3'-dideoxy-α-D-ribofuranosyl) adenine

9- (3'-azido-2 ', 3'-dideoxy-aD-ribofuranosyl) adenine was prepared in two steps from N ₆ -octanoyladenine (2.0 g, 7.7 mmol) and 3'-azido-3' deoxythymidine (1.13g, 4.2 mmol) was prepared by the method described by M. Imazawa and F. Eckstein (J. Org. Chem., 43, 3044 [1978]). M.p. = 120 ⁰ C to 122 ° C

UV pH 1Amax258nm \ min 230nm pH 13Amax 260nmAmin 229nm Analysis for C ₁₀ H ₁₂ N ₈ O ₂ :

Calculated: C 43.48%, H 4.38%, N 40.56%; Found: C43.28%, H 4.45%, N 40.38%. ,

Example 39 9 '- (3'-azido-2', 3'-dideoxy-β-D-ribofuranosyl) adenine

9- (3'Azido-2 ', 3'-dideoxy- / 3-D-ribofuranosyl) adenine was prepared in two steps from N ₆ -octanoyladenine (2.0 g, 7.7 mmol, and 3'-azido-3'). deoxythymidine (1.13 g, 4.2 mmol) according to the method of M. Imazawa and F. Eckstein, J. Org. Chem., 43, 3044 (1978) Melting point = 184 ° C to 185 ° C

 UVpH Umax257nmAmin 230nm, '

pH 13Amax 260nmAmin 228nm Analysis for C ₁₀ H ₁₂ N ₈ O ₂ :

Calculated: C43.48%, H4.38%, · N40.56%; Found: C43.33%, H 4.45%,. N 40.41%.

Example 40 S'-acetyl-S-azido-S-benzoyl-S-deoxythymidine

5'-Acetyl-3'-azido-3'-deoxythymidine (0.75 g, 2.4 mmol) was dissolved in pyridine (5 mL) and benzoyl chloride (1.4 mL, 12 mmol, 5 equivalents) was added at room temperature. The reaction mixture was stirred overnight and then poured into ice-water (250 ml). The pH of the aqueous solution was adjusted to 1. The product was extracted with chloroform, the organic phase washed with water, dried over MgSO ₄ and filtered. The chloroform was removed and the oily product was rapidly chromatographed on silica gel and eluted with chloroform. The product was recovered as an oil.

H ¹ NMR (DMSO-ds): δ 8.04-7.50 (m, 6H; 3N-benzoyl and 6H), 6 6.12 (dd, 1H, J _r , _2a . = 5.6Hz, J, -, ₂ b · = 6.7 Hz, VH), 8 4.55-3.96 (m, 4H, 3Ή, 4Ή, 5 H '), δ 2.62-2.38 (m, 2H, 2' H), δ 2.07 (s, 3H, 5'-acetyl CH ₃ ), δ 1.90 (d, 3 H, J ₅ , ₆ = 1.0 Hz, 5CH ₃ ).

Analysis for C ₁₉ H ₁₉ N ₅ O ₆ :

Calculated: C55.20%, H4.63%, N 16.94%;

Found: C55.29%, H4.64%, N 16.93%. '

Example 41 Threo-3'-azido-5-bromo-2 ', 3'-dideoxyuridine

Threo'-3'-azido-5-bromo-2 ', 3'-dideoxyuridine was prepared from 2'-deoxyuridine in a five-step reaction sequence. The protection of the 5'-hydroxyl group of 2'-deoxyuridine with a Triphenylrnethylgruppe was carried out in a conventional manner. The 3'-hydroxyl was mesylated. By the addition of 2'-deoxy-3'-mesyl-5'-trityluridin (22g, 4OmMoI) to a solution of sodium azide (7,84g, 12OmMoI, 3 equivalents) in dimethylformamide (380ml) at 80 ⁰ C was added a 3 ' -Azido group introduced with the correct stereochemistry. The reaction was continued for 35 hours. The solution was poured into ice-water (2 liters) and the precipitate collected by filtration. The product was isolated in 51% yield by chromatography on silica gel, eluting with chloroform: methanol (1: 1 v / v). The 5'-hydroxylation was deblocked with 80% acetic acid on a steam bath over a period of 25 minutes. After cooling, the tritylcarbinol was filtered off. The filtrate was concentrated in vacuo to a thick oil. The product was isolated by flash chromatography on silica gel, eluting with chloroform: methanol (85:15 v / v). The 5-position of the threo-3'-azido-2 ', 3'-dideoxyuridine was brominated according to exactly the same procedure as given for the bromination of deserythro-3'-azido-2', 3'-dideoxyuridine

 UVpH Umax 280, = 9400, amine 244, = 2600 pH 13Amax 276 = 6700, amine 251 = 3700

H ¹ NMR (DmSo-d ₆ ): δ 11.86 (s, 1H, 3-NH), δ 8.02 (s, 1H, 6H), δ 5.99 (dd, 1H, J _{r > 2a} . = 3.0Hz;

J _r , _2b . = 7.5Hz, 1 Ή), δ 5.1 (t, 1 H, J _{5, CH} 2 5'OH = 5.4Hz, 5ΌΗ), δ 4.49 (m, 1 H, 3'H) ', δ 4.05 (m, 1H, 4'H), δ 3.71 (m, 2H, 5'H), δ 2.72 (m, IH, 2b '), δ 2.18 (m, TH , 2a ')

Analysis for C ₉ H ₁₀ BrN ₅ O ₄ .0.25H ₂ O. 0.1C ₂ H ₄ O ₂ :

Calculated: C 32.25%, H 3.21%, N 20.44%, Br 23.32%; ,

Found: C 32.17%, H 3.21%, N 20.33%, Br 23.19%.

Example 42 1- (3'-azido-2 ', 3'-dideoxy- / 3-D-erythro-pent'ofuranosyl) -2- (benzyloxo) -5-methyl-4- (1H) -pyrimidine (0.4 g, 17.4 ml of vinyl ether, 2.6 equivalents of dry benzyl alcohol (10 ml) for 1 hour at room temperature.) 2,5'-O-Anhydro-3'-azido-3'-deoxythymidine (1.65 g, 6 The reaction was allowed to proceed for 1 hour After pouring into ice-water (250 ml), the pH was adjusted to 7 and the aqueous phase extracted with ethyl acetate The organic phase was extracted with water (4 times). After drying over MgSO ₄ , the ethyl acetate was removed in vacuo and the resulting oil was chromatographed on silica gel, eluting first with ethyl acetate and then with ethyl acetate: methanol (9: 1 v / v) the solvents were removed in vacuo to give an oil, and the oil crystallized after covering with ethyl ether, mp = 125 ° C to 126, 5 ° C

 UV pH 1 unstable

pH 13max256, = 10700, amine 240, = 8900.

H ¹ NMR (DMSO-d ₆ ) -6 7.8 (d, 1H ₁ J ₆₅ = 1.2Hz, 6H), δ7.49-7.38 (m, 5H, 2-phenyl), 6 6.08 (dd, 1 H, J _r , _2a . = 5.0Hz, J ₁ , _2b - = 7.0Hz, TH), 05.37 (s, 2H, 2CH ₂ ), δ 5.25 (t, 1H , J _{5, C} h ₂ 5oh = 5.4Hz, 5'OH), δ4.36-4.32 (m, 1H, 3'H), δ3.85-3.81 (m, 'i H, 4'H), δ 3.7-3.58 (m, 2H, 5'H), δ 2.53-2.34 (m, 2H, 2'H), δ 1.82 (d, 3H , J ₅ , ₆ = 1.0Hz, 5CH ₃ )

Analysis for C ₁₇ H ₁₉ N ₅ O ₄ :

Calculated: C57.14%, H 5.36%, N 19.60%; Found: C57.02%, H 5.43%, N 19.53%.

Example 43

1- (3'-azido-2 ', 3'-dideoxy- / 3-D-threo-pentofuranosyl) -2-ethoxy-5-methyl-4- (1H) -pyrimidinone threo-3'-azido-5 '-O-mesylthymidine (1.08 g, 3.13 mmol) (prepared from threo-3'-azidothymidine) was dissolved in 100 mL EtOH and treated with NaHCO ₃ (0.26 g, 3.13 mmol) at reflux for 18 hours. The reaction mixture was cooled and filtered.

The solvents were removed in vacuo and the residue was applied to a silica gel column followed by elution with CHCl ₃ / MeOH (9: 1 v / v). Combination of the appropriate fractions and removal of the solvents in vacuo gave 0.7 g (2.4 mmol, 75.7%). M.p. = 120 ⁰ C to 122 ° C.

UV (nm): at pH 1 Amax = 260 (ε = 9300), amine = 237 (ε = 5500), ASchulter - 221 "(ε = 7 500), at pH 13 Amax = 256 (ε = 10000), amine = 240 (ε = 7700).

H ¹ NMR (DMSO-d ₆ ): 87.58 (s, 1H, H6), 86.0 (dd, 1H, H1 ', J = 2.9.4, 56Hz), 85.06 (t, 1 H, 5'OH, J = 4.91 Hz), 84.51-4.47 (m, 1H, H 3 '), 84.34 (q, 2H, -OCH ₂ -, J = 7.14Hz , 84.10-4.05 (m, 1H, H4 '), S3.73 (t, 2H, H5', J = 5.62Hz), 82.82-2.73 (m, 1H, H2'b), 82.21 to 2.14 (m, 1H, H2'a), 81.82 (s, 3H, 5CH _3), 81.31 (t, 3H, -CH ₂ -CH _3, J = 6.65Hz).

Analysis for Ci ₂ H ₁₇ N ₅ O ₄ :

Calculated: C 48.81%, H 5.80%, N 23.72%;

Found: C48.59%, H 5.86%, N 23.64%.

Example 44 Threo-3'-azido-2 ', 3'-dideoxy-4-thiothymidine

Threo-3'-azido-5'-O-trityl-3'-deoxy-4- (1,2,4-triazole) -thymidine (1.25 g, 2.2 mmol) (prepared by the method of W.Sung Nucleic Acids Research [1981] 9, 6139) was dissolved in 100 ml of acetone and 30 ml of H ₂ O and then treated with 0.22 g of NaSH-xH ₂ O. The mixture was stirred for 3 hours. The volume was brought to half and extracted with 300 ml CHCl ₃ . The CHCl ₃ was washed with 50 ml of H ₂ O, dried over Na ₂ SO ₄ , and, after removal of the CHCl ₃ in vacuo, gave an oil. The 5'-O-trityl group was removed by dissolving this oil in 100 ml of 80% HOAc. The solution was heated on a steam bath for 2 hours and then cooled, diluted with 100 ml of water and filtered. The solvents were removed in vacuo and the oil applied to a silica gel column. Elution was carried out with CHCl ₃ / MeOH (20: 1 v / v). The appropriate fractions were collected and then the solvents were removed in vacuo. O, 18g (0.62 mmol, 28%) was obtained.

 Melting point = 65 ° C to 67 ° C

 UV (nm): at pH 1 Amax = 337 (ε = 20,700), amine = 280 (ε = 1,200), ACHulter = 238 (ε = 3400); at pH 13Amax = 320 (ε = 18400), amine = 257 (ε = 1700).

H ¹ NMR (DMSO-d ₆ ): 87.63 (s, 1H, H1 ', J = 2.93,4.89Hz), 85.06 (s, 1H, 5ΌΗ), 84.50-4 , 46 (m, 1H, H3 '), 84.10-4.04 (m, 1H, H4'), 83.73 (d, 2H, H5 ', J = 5.61 Hz), 82, 78 to 2.68 (m, 1H, H2'b), 82.20 to 2.14 (m, 1H, H2'a), 82.00 (s, 3H, 5CH _3). Analysis for Ci ₀ H ₁₃ N ₅ O ₃ S · 0.1 C ₂ H ₆ O · 0.25H ₂ O: Calculated: C 41.90%, H 4.86%, N.23.95%, S 10, 97%; Found: C 41.99%, H4.73%, N23.88%, S10.91%.

Example 45 4-Amino-3'-azido-5-bromo-2 ', 3'-dideoxyuridine

3'-azido-2 ', 3'-dideoxyuridine was acetylated according to the method of Visser (Synthetic Procedures in Nucleic Acid Chemistry, Vol.1, page 410) and brominated to give 5'-acetyl-3'-azido-5 bromo-2 ', 3'-dideoxyuridine. This material was reacted with 5 equivalents of 1,2,4-triazole and two equivalents of 4-chlorophenyldichlorophosphate in dry pyridine at room temperature for 7 days to give 5'-acetyl-3'-azido-5-bromo-4- (1.2 , 4-triazolyl) -2 ', 3'-dideoxyuridine as a yellow oil in moderate yield. The treatment at room temperature with methanol, which had been saturated with ammonia at 0 ⁰ C, for a period of 18 hours gave, after recrystallization from ethyl acetate and filtration of the crystals of 4-amino-3'-azido-5-bromo-2 obtained '3'-dideoxyuridine in a yield of 173 mg (0.5 mmol, 6.3%). M.p. = 162 ⁰ C to 165 ⁰ C (dec.)

 UV (nm): at pH 1 Amax = 300.215 (ε = 10700, 12100), amine = 253 (ε = 1500); at pH 13 Amax = 288 (ε = 7300) amine = 260 (ε = 3900).

H ¹ NMR (DMSO-d ₆ ): 88.3 (s, 1H, H ₆ ), 87.85 (broad, 1H, 4- ^j NH ₂ ), 87.05 (broad, 1H, 4-NH ₂ ), S6; 0 (t, 1H, H1 ', J = 5.94Hz), 85.33 (t, 1H, 5'-OH, J = 5.01Hz), 84.4-4.3 (m, 1 H, H3 '), 83.9-3.5 (m, 3H, H4', H5 '), 82.31 (t, 2H, H2', J = 6.27Hz). Analysis for C ₉ HnN ₆ O ₃ Br:

Calculated: C 32.64%, H 3.35%, N 25.38%, Br 24.13%; Found: C32.52%, H3.41%, N 25.32%, Br24.04%.

Example 46 3'-azido-5-bromovinyl-2 ', 3'-dideoxyuridine

5-Bromovinyl-2'-deoxyuridine (BVDU) was synthesized in similar yields using the method of Jones et al., Tetrahedron Letters, 45, 4415 (1979). BVDU was tritylated and mesylated according to the method of Horwitz et al., J. Org. Chem., 31, 205 (1966). This product was refluxed with an equimolar amount of sodium bicarbonate in methanol to give 3 ', 2-O-anhydro-5-bromovinyl-2'-deoxyuridine. This product was treated with 3 equivalents of lithium azide in 1% water / dimethylformamide at 130 ° C. Silica gel chromatography of the crude followed by combination and evaporation of the appropriate fractions afforded 3'-azido-5-bromovinyl-2 ', 3'-dideoxyuridine as a gold oil.

UV (nm): at pH 1 Amax = 292.247 amine = 270.238;

at pH 13 Amax = 253, amine = 238, Ash = 284.

H ¹ NMR (DMSO-d ₆ ): 88.06 (s, 1H, H ₆ ), 87.25 (d, 1H, -CH = CHBr, J = 13.4Hz), π 6.85 (d, 1 H, = CHBr, J = 13.7Hz), 86.07 (t, 1 H, H ₁ -J = 6.3Hz), 65.30 (broad s, 1 H, 5'-OH), 84, 42 (q, 1 H, H ₃ , J = 6.3, 1, 6 Hz), 83.86-3.82 (m, 1 H, H ₄ -), 83.68-3.59 (m , 2H, H ₅ ·), 32.47-2.32 (m, 2H, H _2. ).

Calculated: C 36.89%, H 3.38%, N 19.55%; Found: C 36.86%, H 3.41%, N 19.51%.

Example 47 Threo-3'-azido-5-chloro-2 ', 3'-dideoxyuridine

The title compound was prepared in an analogous manner as 3'-azido-5-chloro-2 ', 3'-dideoxyuridine (Example 31) to yield 0.15 g (0.5 mmol, 25%). Melting point = 65 ° C

 UV (nm): at pH 1 Amax = 278.212 (ε = 8900), amine = 240 (ε = 1600);

at pH 13 Amax = 275 (ε = 6600), amine = 248 (ε = 3300).

H ¹ NMR (DMSO-d ₆ ): 811.89 (s, 1 H, NH), 87.94 (s, 1 H, H ₆ ), 86.00 (dd, 1 H, H _r , J = 2 , 93, 4.64Hz), 85.1 (t, 1H, 5'OH, J = 5.1Hz), 84.50-4.46 (m, 1H, H ₃ -), 84.08 -4.03 (m, 1H, H ₄ -), 83.71 (t, 2H, H _5. , J = 5.32 Hz), 82.77-2.67 (m, 1H, H ₂ , b), 82, 23-2, 16 (m, 1 H, H ₂ , a).

Calculated: C37.85%, H3.72%, N23.48%, Cl 11.89%; Found: C37.94%, H 3.91%, N23.23%, Cl 11.86%.

Example 48 1- (3'-azido-2 ', 3'-dideoxy- ^ 3-D-erythro-pentofuranosyl!) - 5-methyl-2- (pentyloxo) -4- (1H) -pyrimidinone 1- (3 '-Azido-2', 3'-dideoxy-.beta.-D-erythro-pentofuranosyl) -5-methyl-2- (pentyloxo) -4- (1H) -pyrimidinone was obtained by the method used to prepare 1 - (3'-azido-2 ', 3'-dideoxy-0-D-erythro-pentofuranosyl) -2- (benzyloxo) -5-methyl-4- (1H) -pyrimidinone was used (Example 42). The final purification was achieved by HPLC on C _18, eluted with water: methanol (3: 7). The solvents were evaporated and the product obtained as an oil.

UVpH 1 Amax 258nm = 9400, amine 232nm = 5900 pH 13 Amax 256nm = 10600, amine 239nm = 8200

NMR was performed in DMSO-d ₆ .

NMR: 87.81 (s, 1H, 6H), 86.04 (t, 1H, J _r , ₂ x = 6.7Hz, 1Ή), 35.28 (t, 1H, J ₅ , CH ₂ , ₅₀ h = 5.1 Hz, 5'OH), 84.43-4.37 (m, 1H, 3'H), 84.27 (t, 2H, J ₂ -O (I ₂ 2CH ₂ ) = 6.6Hz, 2-0 (CH ₂ J ₁ ), 33.88-3.84 (m, 1K, 4'H), 83.70-3.50 (m, 2H, 5'CH ₂ ), 82.50-2.34 (m, 2H, 2'H) -, 31.80 (s, 3H, 5CH ₃ ), 81.72 - 1.68 (m, 2H, 2-O (CH ₂ ) ₂ ), 81.38-1.30 (m, 4H, 2-0 (CH ₂ I ₃ and ₄ ), 80.92-0.87 (m, 3H, 2-O (CH ₃ ) ₅ ).

Analysis for C ₁₅ H ₂₃ N ₅ O ₄ - ¹ AH ₂ O:

Calculated: C 52.70%, H 6.93%, N 20.48%;

Found: C52.63% H 6.92%, N 20.48%.

Example 49 3'-Azido-3-benzoyl-3'-deoxy-5'-mesylthymidine

3'-Azido-3-benzoyl-3'-deoxy-5'-mesylthymidine was prepared from 3'-azido-3'-deoxy-5'-rnesylthymidine by a method analogous to that used in Example 40 to produce 6'-acetyl -S'-azido-S-benzoyl-S'-deoxythymidine prepared from 5'-acetyl-3'-azido-3'-deoxythymidine

Melting point = 86 ⁰ C to 88 ° C.

UVpH 1 Amax 259 nm = 21200, amine 230nm = 6400 pH 13Amax265nm = 9600, amine 248nm = 8000.

H ¹ NMR (DMSO-d ₆ ): 88.00-7.58 (m, 6H, 6H and phenyl), 86.15 (t, 1 H, J _v , _2. = 6.1 Hz, VH), 84.55-4.47 (m, 3H, 3Ή and 5'CH ₂ ), 84.12-4.10 (m, 1H, 4'H), 83.28 (s, 3H, 5'-SO ₂ CH ₃ ), 82.64-2.4 (m, 2H, 2'H), 81.89 (d, 3H, J ₅ , ₆ = 1.3Hz, 5CH ₃ ). Analysis for Ci ₈ H ₁₉ N ₅ O ₇ S:

Calculated: C48.10%, H4.26%, N15.58%, S7.13%; Found: C48,00%, H4,23%, N 15,39%, S7,10%.

Example 50 Threo-3'-azido-2 ', 3'-dideoxy-5-iodouridine

5-iodo-2'-deoxyuridine was tritylated and mesylated according to Horwitz et al., J. Org. Chem., 31, 205 (1966). This product was heated with 3 equivalents of lithium azide in anhydrous dimethylformamide at 74 ° C for 48 hours. Silica gel chromatography of the reaction mixture with 20: 1 CHCl ₃ Z ¹ MeOH (v / v) followed by combining and evaporating the appropriate fractions afforded threo-3'-azido-2 ', 3'dideoxy-5-iodo 5'-trityluridin. The 5'-hydroxyl was deblocked with a saturated bromide / nitromethane solution at 0 ⁰ C. Silica gel chromatography of the crude product using CHCl ₃ / MeOH (9: 1, v / v) followed by combining and evaporating the appropriate fractions afforded threo-3'-azido-2 ', 3'-dideoxy-5- ioduridine as a solid. Melting point = 80 ^° C.

UV (nm): at pH 1 amine = 247, at pH 13 Amax = 277, amine = 252.

H ¹ NMR (DMSO-d ₆ ): 88.37 (s, 1 H, H 6), 86.00 (t, 1 H, H1 ', J = 6.17 Hz), 85.4-5.3 (m, '1H, 5'OH), 84.4-4.3 (m, 1H, H 3'), 83.9-3.8 (m, 1H, H4 '), 83.7 -3.6 (m, 2H, H 5 ').

Analysis for C ₉ H ₁₀ N ₅ O ₄ J · 0.4C ₄ H ₈ O ₂ :

Calculated: C30.73%, H3.21%, N 16.90%, J30.63;

Found: C30.93%, H3.09%, N 16.61%, J30.94%.

-27- ZOΊ S84 Example 51

6'-Acetyl-S'-azido-S'-deoxythymidine was reacted with 5 equivalents of 1,2,4-triazole and two equivalents of 4-chlorophenyldichlorophosphate in dry pyridine at room temperature for a period of 10 days. Silica gel chromatography of the crude product using 1: 1 EtOAc / hexane (v / v) followed by combining and evaporation of the appropriate fractions afforded an oil. Crystallization from EtOAc afforded the title compound as a solid in an amount of 2.7 g (7.5 mmol, 60%). Melting point = 143 ⁰ C to 145 ⁰ C.

 UV (nm): at pH 1 Amax = 324.245.215 (ε = 9300, 10,000, 20500), ληιίη = 282.233 (ε = 2100.8200); at pH 13 Amax = 276 (ε6000), amine = 242 (ε = 2000).

H ¹ NMR (DMSOd ₆ ): 69.34, 8.40 (2s, 2H, triazolyl), 68.23 (s, 1H, H6), 66.12 (t, 1H, H V, J = 6 , 16Hz), 64.48-4.17 (m, 4H, H3 ', H4', H5 '), 62.35 (s, 3H, 5'-acetyl), 62.07 (s, 3H, 5CH ₃ )

Analysis for C ₁₄ H ₁₆ N ₈ O ₄ : Calculated: C 46.67% H 4.48%, N 31.10%

 Found: C46.58%, H 4.51%, N 31.02%.

Example 52 1- (3'-azido-2 ', 3'-dideoxy-1-D-erythro-pentofuranosyl) -4-dimethylamino-5-methyl-2- (1H) -pyrimidinone 5'-acetyl-3' Azido-3'-deoxy-4- (1,2,4-triazolyl) -thymidine was dissolved in dry acetonitrile at room temperature under a nitrogen atmosphere. Dimethylamine, 20 equivalents, was added all at once and the reaction mixture was stirred for 30 minutes. The solvents were removed, the residue dissolved in methanol saturated with ammonia and the solution stirred for 30 minutes. The solvents were removed and the residue was dissolved in EtOAc. The title compound precipitated out of the solution slowly and was filtered off to give a white solid. Melting point = 157 ° C to 159 ° C

 UVfnm): at pH 1 Amax = 299.244 (ε = 14900, 7900), Xmin = 254 (ε = 2500); at pH 13 Amax = 287 (ε = 14,000), amine = 243 (ε = 6800).

H ¹ NMR (DMSO-d _e ): 67.63 (s, 1 H, H 6), 66.06 (t, 1 H, H V ₁ J = 6.53 Hz), 65.22 (t, 1 H, 5'OH, J = 5.2Hz), 64.37-4.34 (m, 1H, H3 '), 3.83-3.80 (m, 1H, H4'), 63.65-3 , 60 (m, 2H, H 5 '), 63.06 (s, 6H, N (CH ₃ J ₂ ), 62.29-2.23 (m, 2H, H2'), 52.11 (s , 3H, 5-CH ₃ ) Analysis for C ₁₂ H ₁₈ N ₆ O ₃ :

Calculated: C, 48.97%, H, 6.16%, N, 28.56%; Found: C 49.06%, H 6.20%, N 28.50%.

Example 53 1- (3'-azido-2 ', 3'-dideoxy- / S-D-erythro-pentofuranosyl) -5-methyl-2- (isopentyloxo) -4- (1H) -pyrimidinone

The title compound was prepared by an analogous procedure to that for the preparation of 1- (3'-azido-2 ', 3'-dideoxy- / 3-D-erythropentofuranosyl) -2- (benzyloxo) -5-methyl-4- (1 H) -pyrimidinone (Example 42). UV (nm): at pH 1 Amax 258 = 9000, amine 237 = 6000

 at pH 13 Amax 255 = 11,000, amine 239 = 8,000.

H ¹ NMR (DMSO-d ₆ ): 67.79 (s, 1H, 6H), 66.02 (t, 1H, J _r , ₂ x = 6.3Hz, VH), 6 5.23 (t , 1 H, J ₅ oh, s'h = 5.2Hz, 5ΌΗ), 64.4-4.3 (m, 3Ή and 2-0-CH ₂ -CH ₂ CH (CH ₃ I ₂ ), 63, 9-3.8 (m, 1H, 4'H), 63.7-3.5 (m, 2H, 5'H), 62.5-2.3 (m, 2'H), 61 , 78 (s, 3H, 5CH ₃ ), 61.7-1.5 (m, 3H, 2-OCH ₂ -CH ₂ -CH (CH ₃ I ₂ ), 60.92 and 0.89 (d, 6 H, J = 6.3 Hz, 2-O (CH ₂ ) ₂ CH (CH ₃ ) ₂ ) Analysis for C ₁₅ H ₂₃ N ₅ O ₄ :.

Calculated: C53.40%, H 6.87%, N 20.76%; Found: C53.14%, H 6.92%, N 20.62%.

Example 54 S-Acetyl-S'-azido-S'-O-f S -chlorobenzo O-S'-deoxythymidine

To a mixture of 3'-azido-5'-O- (4-chlorobenzoyl) -3'-deoxythymidine (0.3g, 0.74mmol) and silver cyanide (0.4g, 3, OmMoI) in 20ml benzene was added acetyl chloride ( 2.6 g, 33 mmol) in excess in several portions. The mixture was stirred at room temperature until the starting material disappeared (4 hours) as determined by TLC CHCl ₃ / MeOH (20: 1, v / v). The reaction mixture was filtered and the filtrate evaporated to dryness under reduced pressure. The resulting oily residue was chromatographed on silica gel and eluted with chloroform / hexane (1: 1, v / v) followed by chloroform to give 0.21 g (64%) of the desired product as an oil. UV (nm): at pH 1 Amax 273 (ε = 9000), amine 251 (ε = 6200); at pH 13 Amax 266 (ε = 8800), amine 247 (ε = 7000).

H ¹ NMR (DMSO-d ₆ ): 61.68 (s, 3H, 5-CH ₃ ), '62.47 (s, 3-COCH ₃ ), 62.3-2.5 (m, 2'H ), 64.1-4.2 and 4.4-4.7 (m, 4H, 3'H, 4'H and 5'H), 66.1 (t, 1 H, J _{n 2.} = 6 , 3Hz, VH), 67.5-8.0 (m, 5H, 6H and phenyl)

Analysis for C ₁₉ H ₁₈ N ₅ O ₆ CI -0.2CHCl ₃

Calculated: C, 48.89%, H, 3.89%, N, 14.85%, Cl, 12.03%; Found: C 49.05%, H 3.94%, N 14.79%, Cl 12.56%.

Example 55 3'-azido-5-cyano-2 ', 3'-dideoxyuridine

5-iodo-2 ', 3'-dideoxyuridine was added to 2', 3'-dideoxy-3 ', 5'-diacetyl-5 with acetic anhydride (2.1 equivalents) in pyridine with exclusion of moisture at room temperature for 2 hours. ioduridine reacted. The nucleoside (1 equivalent), potassium cyanide (1.3 equivalents) and potassium acetate (1.3 equivalents) were heated together in dry DMSO for 2 hours under nitrogen to 97 ^° C. The solvents were removed in vacuo and the residual oil applied to a silica gel column followed by elution with CHCl ₃ / EtOAc (1: 1, v / v). The appropriate fractions were collected, combined, and, after evaporation, gave 5-cyano-2 ', 3'-dideoxy-3', 5'-diacetyluridine. The compound was dissolved in methanol saturated with ammonia and stirred at 0 ^° C for 18 hours. The solvents were removed in vacuo at room temperature to give the title compound as crystals. Melting point = 160 ° C to 162 ° C.

UV (nm): at'pH 1 Amax = 276.215 (ε = 13500.1'1 200), amine = 238 {ε = 1700); at pH 13Amax = 276 (ε = 1010O) ₇ AmIn = 240 (ε = 3400).

H ¹ NMR (DMSO-d ₆ ): 58.81 (s, 1H, H6), 56.00 (t, 1H, H V, J = 5.94Hz), 55.3-5.21 (m , 2H, 5ΌΗ, 3ΌΗ), 54.28-4.15 (m, 1H, H3 '), 53.82-3.77 (m, 1H, H4'), 53.7-3.5 (m, m, 2H, H5 '), 52.18 (t, 2H, H 2', J = 5.75 Hz)

 AIfUCHNOO ^ SHO

Calculated: C 46.61%, H4.50%, N 16.31%;

Found: C46.67%, H4.71%, N16.54%.

Example 56

1- (3'-azido-2 ', 3'-dideoxy- / 3-D-threo-pentofuranosyl) -5-methyl-2-pentyloxy-4- (1H) -pyrimidinone 2,5'-O-anhydro -1- (3'-azido-2 ', 3'-dideoxy- / 3-D-threo-pentofuranosyl) thymine was added to a solution of potassium t-butoxide (0.5 equivalent) in pentan-1-ol added. The reaction mixture was stirred for 2 hours at room temperature under nitrogen. The solvents were removed in vacuo and the residue was applied to a silica gel column. Elution with CHCl ₃ / MeOH (20: 1, v / v) and subsequent combination and evaporation of the appropriate fractions afforded a clear oil, which on standing afforded slowly crystals of the title compound. Melting point = 110 ⁰ C to 111 "C.

UV (nm): at pH 1 Amax = 255 (ε = 10200), amine = (ε = 2600), Ash = 222 (ε = 9000); at pH 13 Amax = 251.226 (ε = 11,600, 9600), amine = 238 (ε = 8600).

H ¹ NMR (DMSO-d ₆ ): 57.58 (s, 1H, H6), 55.98 (dd, 1H, HV, J = 2.98, 4.88Hz), 85.07 (t, 1H, 5'OH, J = 5.42Hz), 54.5-4.47 (m, 1H, H 3 '), 54.31 ^ 1.25 (m, 2H, -OCH ₂ -), 54.1-4.06 (m, 1H, H4 '), 53.73 (t, 2H, H 5', J = 5.62Hz), 52.82-2.72 (m, 1H, H 2 '), 52.2-2.14 (m, 1 H, H 2'), 51.82 (s, 3H, 5-CH ₃ ), δ 1.75-1.65 (m, 2H, Pentyl), δ 1.4-1.3 (m, 4H, pentyl), 50.92-0.87 (m, 3H, pentyl). , Analysis for C ₁₅ H ₂₃ N ₄ O ₄ :

Calculated: C53.40%, H6.87%, N 20.76%; Found: C 53.31%, H6,90%, N20,74%.

Example 57

1- (3'-azido-2 ', 3'-dideoxy- / 3-D-threo-pentofuranosyl) -2-benzyloxy-5-methyl-4- (1H) -pyrimidinone

The title compound was obtained in an analogous manner as described for the preparation of 1- (3'-azido-2 ', 3', dideoxy- / 3-D-threo-pentofuranosyl) -5-methyl-2-pentyloxy-4- (1H ) -pyrimidinone (Example 56) except that benzyl alcohol was used instead of pentan-1-ol. Melting point = 137 ⁰ C to 139 ° C.

 UV (nm): at pH 1 Amax = 266 (ε = 9100), amine = 235 (ε = 3000); at pH 13 Amax = 256 (ε = 11,500), amine = 240 (ε = 9600).

H ¹ NMR (DMSO-de): δ7.6 (s, 1H, H 6), 57.49-7.37 (rri, 5H, phenyl), 56.01 (dd, 1H, H 1 ', J = 2.52.5.OHz), 55.35 (s, 2H, benzyl), 55.1-5.0 (m, 1H, 5ΌΗ), 54.5-4.4 (m, 1H, H3 '), 54.1-4.0 (m, 1H, H4'), 53.77-3.67 (m, 2H, H5 '), 82.85-2.70 (m, 1H, H2 '), 52.25-2.15 (m, 1H, H 2'), 51.83 (s, 3H, 5-CH ₃ )

Analysis for C ₁₇ H ₁₉ N ₅ O ₄ 0.25H ₂ O: Calculated: C 56.43%, H 5.43%, N 19.35%; Found: C56.51%, H 5.37%, N 19.36%.

Example 58

1- (3'-azido-2 ', 3'-dideoxy-β-D-erythro-pentofuranosyl) -4-methoxy-5-methyl-2 (1H) -pyrimidinone

5'-Acetyl-3'-azido-3'-deoxy-4- (1,2,4-triazolyl) -thymidine was treated with 0.5 N sodium methoxide in methanol for 24 hours at room temperature. The solution was neutralized to pH 7 with a sulfonic acid resin (DOW 5OW, H ⁺ ), the resin filtered off and the filtrate evaporated in vacuo to a solid. The solid was dissolved in a small amount of CHCl ₃ , applied to a silica gel column and eluted with 2% MeOH in CHCl ₃ . The appropriate fractions were collected, combined and evaporated to dryness to give a white solid. Melting point = 119 ^° C. to 123 ^° C. 'UV (rim): at pH 1 Amax = 279 (ε = 7500), amine = 239 (ε = 1700); at pH 13 Amax = 279 (ε = 6400), amine = 243 (ε = 1300).

H ¹ NMR (DMSO-d ₆ ): 58.02 (s, 1H, H6), 86.08 (t, H, H V, J = 5.86Hz), 55.29 (t, 1H, 5 'OH, J = 5.48Hz), 54.41 ^, 33 (m, 1H, H3'), ri3.9-3.86 (m, 1H, H4 '), 53.86 (s, 3 H, 4-OCH ₃ ), 53.75-3.58 (m, 2H, H 5 '), 52.4-2.3 (m, 2H, H2'), 51.89 (s, 3H, 5-CH ₃ ). Analysis for C ₁₁ H ₁₅ N ₅ O ₄ :

Calculated: C 46.97% ,. H 5.38%, N 24.90%; Found: C47.06%, H 5.40%, N 24.86%.

Example 59.

1- (3'-azido-2 ', 3'-dideoxy-.beta.-D-erythro-pentofuranosyl) -5-methyl-4- (1-pyrrolidinyl) -2- (1H) -pyrimidinone

5'-Acetyl-3'-azido-3'-deoxy-4- (1,2,4-triazolyl) -thymidine was dissolved in dry acetonitrile at room temperature under a nitrogen atmosphere. Pyrrolidine (5 equivalents) was added dropwise over 5 minutes and the reaction mixture was stirred for 2 hours. The solvents were evaporated in vacuo to give an oil. The oil was dissolved in methanol saturated with ammonia at room temperature and stirred for 4 hours. The solvents were removed in vacuo and the residue was applied to a silica gel column. Elution with CHCl ₃ / MeOH (20: 1, v / v) followed by combining and evaporation of the appropriate fractions afforded the title compound as a white solid. Melting point = 168 ⁰ C to 171 ° C

 UV (nm): at pH 1 Amax = 295, 233 (ε = 15700, 9500), amine = 253 (A = 2600); at pH 13Amax = 285 (ε = 15300), amine = 241 (ε = 7900).

H ¹ NMR (DMSO-de): 57.57 (s, 1H, H6), 56.02 (t, 1H, HT, J = 6.5Hz), (t, 1H, 5'OH, J = 15Hz), 54-4.3 (m, 1H, H3 '), 83.84-3.77 (m, 1H, H4'), 53.67-3.51 (m, 6H, H 5 ' , 4-pyrrolidin H's), 52.24 (t, 2H, H2 ', J = 6,08Hz), 52.14 (s, 3H, 5-CH _3), 51.87 to 1.78 (m, 4H , Pyrrolidine).

Analysis for Ci ₄ H ₂₀ N ₆ O ₃ :

Calculated: C52.49%, H6.29%, N26.23%;

Found: C 52.37%, H 6.33%, N 26.17%.

Example 60 1- (3'-azido-2 ', 3'-dideoxy-j3-D-erythro-pentofuranosyl) -4-benzyloxy-5-methyl-2- (1H) -pyrimidinone

S'-Acetyl-S'-azido-S'-deoxy-O-Atriazoly-O-thymidine dissolved in dry acetonitrile was added dropwise at room temperature under nitrogen over 5 minutes to a suspension of benzyl alcohol (5 equivalents) and potassium tert-butoxide (2 equiv.) in dry acetonitrile. The reaction mixture was stirred for 1 hour and the solvents removed in vacuo. The residue was placed on a silica gel column and eluted with CHCl ₃ / EtOAc (3: 1, v / v). The appropriate fractions were collected, combined and evaporated to dryness to give a solid. The solid was recrystallized from CH 2 Cl / EtOAc (1: 1, v / v) and the resulting crystals were filtered off to give the title compound. Melting point = 133 ° C to 134 ° C

 UV (nm): at pH 1 Amax = 276, (ε = 8000), amine = 237 (ε = 2000); at pH 13 Amax = 281 (ε = 8100), amine = 237 (ε = 1600).

H ¹ NMR (DMSOd ₆ ): 5 8.05 (s, 1 H, H 6), 57.45-7.35 (m, 5H, phenyl), 56.07 (t, 1 H, HT, J = 5.94Hz), 55.35 (s, 2H, benzyl), 55.27 (t, 1 Η, δ'ΟΗ, J = 5.20Hz), 54.41-4.31 (m, 1H, H 3 '), 53.91-3.83 (m, 1H, H4'), 53.7-3.6 (m, 2H, H 5 '), 52.4-2.3 (m, 2H, H 2 '), 51.9 (s, 3H, 5-CH ₃ ).

Analysis for C19H21N ₅ O _5:

Calculated: C 57.14%, H5.36%, N 19.60%; Found: C57.06%, H 5.37% ,. N 19.58%.

Example 61 S'-acetyl-S'-azido-S-bromo ^ ', S'-dideoxyuridine

3'-azido-2 ', 3'-dideoxyuridine was acetylated and brominated according to the procedure of Visser, Synthetic Procedures in Nucleic Acid Chemistry, 1, 410 and the title compound was obtained. M.p. = 112 ⁰ C to 114 ° C. UV (nm): at pH 1 Amax = 279, 210 (ε = 9500.10600), amine = 243 (ε = 2000); at pH 13 Amax = 276 (ε = 700), amine = 250 (ε = 4000).

H ¹ NMR (DMSO-d ₆ ): 511.88 (s, 1H, NH), 58.02 (s, 1H, H6), 56.08-6.01 (m, 1H, H1 ') , 84.47-4.40 (m, 1H, H3 '), 54.27-4.24 (m, 2H, H5'), 54.03-4.00 (m, 1H, H 4 ' ), 82.41-2.34 (m, 2H, H 2 '), 52.09 (s, 3H, acetyl)

Analysis for CnHi ₂ N ₅ O ₅ Br:

Calculated: C35.31%, H3.23%, N 18.72%, Br21.36%; Found: C35.29%, H3.25%, N 18.66%, Br 21.45%

Example 62 1- (3'-azido-2 ', 3'-dideoxy-^-D-threo-pentofuranosyl-5- (trifluoromethyl) -uracil The 5'-hydroxyl group of 5-trifluoromethyl-2'-deoxyuridine was reacted with a triphenylmethyl protected and the 3'-hydroxy! group mesylated. the product (3.0 g, 4.9 mmol) was reacted with 0.7 g lithium azide in 50 ml of DMF over a period of about 26 hours at 70 ⁰ C. the cooled reaction mixture was The solid was isolated, washed with water and purified by flash chromatography using hexane / ethyl acetate (2: 1, v / v), the appropriate fractions were combined and the solvent removed to give 0, received 83g of the tritylated product. This was dissolved in a saturated solution of zinc bromide in acetonitrile, and stirred at 0 ⁰ C overnight After addition of 100 ml of ammonium acetate. (1-molar) was the organic layer separated and evaporated to dryness in vacuo. the residue was by Schnellch purified by using CHCl3 / CH3OH (20: 1, v / v). The fractions were combined and evaporated to dryness in vacuo. The yield of the title compound was 0.25 g, 0.8 mmol, 5.3%. Melting point = 118 ° C to 120X

Analysis for C ₁₀ H ₁₀ F ₃ N ₅ O ₄ : '

Calculated: C37.39%, H 3.14%, N 21.80%, F17.74%; Found: C37.31%, H3, .23%, N 21.75%, F 17.60%.

Example 63 1- (3'-azido-2 ', 3'-dideoxy- / 3-D-threo-pentofuranosyl) uracil

The 5'-hydroxyl group of 2'-deoxyuridine was protected and the 3'-hydroxyl group mesylated. The 3'-mesyl group was shifted by heating in dimethylformamide at 80 ° C for 24 hours with lithium azide (3 equivalents) to inversion of configuration. The reaction mixture was poured into ice-water and the product precipitated. After filtration, the moist product was deblocked. Final purification was by chromatography on silica gel, eluting with chloroform / methanol (95: 5). The appropriate fractions were combined and the solvents removed to give the title compound as a solid. Melting point = 142 ° C to 145 ° C.

Example 64 1- (3'-azido-2 ', 3'-dideoxy- / 3-D-erythro-pentofuranosyl) uracN

The S 'hydroxyl group of 2'-deoxyuridine (30 g, 0.13 mol) was tritylated by the method of Horwitz et al., J. Org. Chem., 31, (1966). The 3'-hydroxyl group (12.6g, 0.027 mol) was chlorinated by the method of Example 62. The dimethylacetamide was removed in vacuo and the thick oil poured into water (500 ml). The product was extracted with ether (3x). The solvent was removed and the resulting oil chromatographed on silica gel, eluting first with dichloromethane and then with 1% methanol in dichloromethane. The product fractions were combined and the solvents removed in vacuo. The 5'-hydroxyl group was deblocked without further purification by heating in 80% acetic acid on the steam bath over a period of 20 minutes. Upon cooling, the tritylcarbinol precipitated and was filtered off. The filtrate was concentrated in vacuo and chromatographed on silica gel, eluting with ethyl acetate. The 3'-chlorine was displaced by heating in HMPA with lithium azide (3 equivalents) at 90 ° C overnight. The reaction mixture was poured into water and extracted with chloroform. The chloroform containing the product was dried with magnesium sulfate. Removal of the chloroform afforded a solid which was crystallized first from ethyl acetate / methanol and then in water. The crystallized solid was dissolved in water and applied to an XAO column. After washing with water, the title compound was eluted with ethanol. The ethanol was removed in vacuo to give a solid. Melting point = 166.5 ° C to 168.5 ° C.

Example 65 1- (3'-A2ido-2 ', 3'-dideoxy-erythro / 3-D-pentofuranosyl-5-ethyl) -uracil

5-chloromercuri-2'-deoxyuridine was prepared from 2'-deoxyuridine (10g, 0.44 mol) by the method of Bergstrom and Ruth, J. Carb. Nucl. and Nucl., 4, 257 (1977). 2'-Deoxy-5-ethyluridine was prepared by the method of Bergstrom et al., J. Am. Chem. Soc., 100, 8106 (1978). The 5'-hydroxyl group was protected by the method of Example 62. The 3'-hydroxyl group was chlorinated and the 5'-hydroxyl group unmasked by the method of Example 64. The 3'-chloro of threo-3'-chloro-2 ', 3'-dideoxyuridine was displaced by inversion of configuration by heating in HMPA with lithium azide (5 equivalents) at 55 ° C for a period of 1 hour. The title compound was purified by chromatography on silica gel, eluting with ethyl acetate. Removal of the solvent from the appropriate fractions provided the desired compound

Mp = 112.5 ⁰ C to 115 ° C.

Example 66 1- (3'-azido-2 ', 3'-dideoxy- / 3-D-threo-pentofuranosyl) -5- (2-bromovinyl) -uracil

5-Bromovinyl-2'-deoxyuridine (BVDU) was synthesized in similar yields by the method of Jones et al., Tetrahedron Letters, 45, 4415 (1979). BVDU was tritylated and mesylated according to the method of Horowitz et al., J. Org. Chem., 31, 205 (1966). This product (4.25 g, 6.5 mmol) was dissolved in 100 mL of DMF containing 0.955 g (19.5 mmol) of lithium azide and heated to 74 ° C for 24 hours. The reaction mixture was poured onto 600 ml of ice with stirring. The resulting solid was isolated, purified by chromatography and obtained as a gum (2.48 g). Treatment by dissolving in 100 ml of 80% acetic acid and heating on a steam bath for a period of 3 hours resulted in the deblocking of the compound. The cooled reaction mixture was diluted with water and filtered to remove tritylcarbinol. The filtrate was concentrated to an oil which was purified by flash chromatography in CHCl ₃ / CH ₃ OH (9: 1, v / v). Combination of the appropriate fractions and removal of the solvent yielded a solid which was recrystallized twice from aqueous methanol. The yield was 0.66 g, 1.8 mmol, 27.7%. M.p. = 165 ⁰ C to 166 ° C. AnBIySeTurC ₁₁ H ₁₂ BrH ₅ O ₄ 0.5H ₂ O:

Calculated: C35.98, H3.57%, N 19.07%, Br 21.85%; Found: C 35.98; H 3.57%, N 19.07%, Br 21.76%.

Example 67 1- (3'-azido-2 ', 3'-dideoxy-β-D-threo-pentofuranosyl!) 5-methylisocvosine

i-iS'-azido ^ '' - dideoxy-jS-D-threo-pentofuranosyD ^ -methoxy-S-methyl-iHt-pyrimidinone (0.5g, 2mmol) was dissolved in a bomb containing 15ml of methanol treated with ammonia was saturated, united. After 6 days at room temperature, the bomb was in

an oil bath for 4 days at 65 ⁰ C heated. The reaction mixture was evaporated to dryness and purified by crystallization 3UsCHCl ₃ ZCH ₃ OH (9: 1, v / v). The solid was washed with CHCl ₃ and dried in air to give 0.16 g, 0.6 mmol (30%) of product.

Melting point = 158 ° C to 160 ⁰ C.

Analysis for C ₁₀ H ₁₄ N ₆ O ₃ · ¹ AH ₂ O:

Calculated: C 44.36%, H 5.40%, N 31.04%:

Found: C44.42%, H 5.36%, N 31.04%.

Example 68 '.

1- (3'-azido-2 ', 3'-dideoxy-.beta.-D-threo-pentofuranosyl) -3-methylthymine

5'-Trityl-3'-threo-3'-azido-3'-deoxythymidine (1.0 g, 1.95 mmol) and Ν, Ν-dimethylformamide dimethyl acetal (Zemlicka, Coll. Czech Chem. Comm., 35, 3572 1972]) (0.93 g, 7.8 mmol) were refluxed in 50 ml of CHCl _{3 for} 96 hours. Removal of the solvent yielded an oil which was further purified by flash chromatography using CHCl ₃ . Removal of the solvent provided 0.54 g of a foam. The deblocking was carried out by heating the foam in 50 ml of 80% acetic acid on a steam bath over a period of 2 hours. Tritylcarbinol was removed by filtration, after which the reaction mixture was diluted with water. The filtrate was concentrated in vacuo to an oil and the oil purified by flash chromatography using CHCl ₃ / EtOAc (2: 1, v / v). The solvent was removed to give the compound as an oil (0.20g). UVmax (nm): beipH 1 Amax = 267 (ε = 8100), Ash = 209 (ε = 8000);

at pH 13 λ max = 267 (ε = 8000)

H ¹ NMR (DMSO-de): 87.56 (s, 1 H, H 6), δ6.07 (dd, 1 H, H ₁ ), 83.17 (s, 3 H, N-CH ₃ ), 81.86 (s, 3 H, 5-CH _3).

Analysis for C ₁₁ H ₁₅ N ₅ O ₄ .0.4HOAc. 0.3H ₂ O: Calculated: C 45.62%, H 5.58%, N 22.54%; Found: C 45.67%, H 5.60%, N 22.57%.

Example 69 (El-S-ti-IS'-azido-a'.S'-dideoxy-Jβ-D-erythro-pentofuranosyl-O-S-SAtetrahydro-C-dioxo-S-pyrimidinyne-acrylic acid 2'-Deoxyuridine converted into the 5-chloromercuri derivative according to the literature procedure (Bergstrom and Ruth, J. Carb. Nucl., 4, 267 [1977]) in 96% yield. This product (50 g, 1.04 mol) was dissolved in 800 ml dry methanol containing ethyl acrylate (104 g, 1.04 mol) and a 0.1 N solution of Li ₂ PdCl ₄ Cl in methanol (1040 ml) The solution was stirred for 4 hours and then treated with hydrogen sulfide for 2 minutes The suspension was then filtered through a celite pad and the filtrate was evaporated to dryness in vacuo The residue was triturated with methanol to give a solid which was filtered off and dried to give 22 g of a white dye tritylated and mesylated by Horowitz et al., J. Org. Chem., 31, 205 (1966). A portion of this material (7.06 g), 10, 9 mmol) was dissolved in 100 ml of dry methanol containing sodium bicarbonate (0.92 g, 10.9 mmol) and heated at reflux for 6 hours. The solvents were removed in vacuo, the residue was vortexed with water, then filtered and dried in air to give 6.1 g of a white dye. This product was dissolved in 50 ml of DMF / 1 ml of water containing lithium azide (1.63 g, 33.2 mmol) and heated to 125 ° C for 4 hours. The reaction product was poured onto 200 ml of ice, the precipitate collected and washed with water. This material was then dissolved in 100 ml of 80% HOAC and heated to 100 ° C for 4 hours. The reaction product was diluted with water and the precipitate filtered off. The filtrate was evaporated to dryness to yield 800 mg of an oil. This oil was dissolved in 20 ml of 0.5N sodium hydroxide solution and stirred for 2 hours at room temperature. The solution was adjusted to pH 3, the precipitate filtered off and dried in air to give 550 mg (1.7 mmol) of the title compound. Melting point> 250 ° C. UV (nm): at pH 1 Xmax = 300 (ε = 20400), Xmin = 230 (ε = 3900), = 262 (ε = 13100); at pH 13 Xmax = 297, 266 (ε = 15600.14800), Xmin = 281-288 (ε = 13600.8600).

H ¹ NMR (DMSO-de): δ 8.37 (s, 1 H, H 6), δ 7.30 (s, 1 H, -CH =, 5 = 15.57 Hz), δ 6.78 ( d, 1 H, = CH-COOH, 5 = 15.93 Hz) δ 6.1-6.06 (m, 1H, HV), δ 5.41-5.37 (m, 1H, 5ΌΗ), δ4,47-4,39 (m, 1H, H3 '), δ3,87-3,83 (m, 1H, H4), δ3,74-3,58 (m, 2H, H 5') , δ 2.54-2.81 (m, 2H, H2 ').

Analysis for Ci ₂ HiSN ₅ O ₆ : Calculated: C 44.59%, H 4.05%, N 21.67; Found: C44.45%, H4.06%, N 21.60%.

Example 70 (EJ-S-ti-tS'-azido) '''-dideoxy-jS-D-threo-pentofuranosyl-1-yl-S-tetrahydro-1'-dioxo-B-pyrimidinyne-2-acrylic acid 2'-deoxyuridine was converted into the 5-chloromercuri derivative in 96% yield according to the literature procedure of Bergstrom and Ruth, J. Carb. Nucl., 4, 257 (1977) This product (50g, 1.04 mol) was dried in 800ml Methanol containing ethyl acrylate (10.4 g, 1.04 mol) and a 0.1 N solution of Li ₂ PdCl ₄ In methanol (10.40 mL) The solution was stirred for 4 hours and then with hydrogen sulfide for 2 minutes The suspension was filtered through a pad of celite and the filtrate evaporated to dryness in vacuo The residue was triturated with methanol to give a solid which was filtered off and dried in air to give 22g of a white solid The method of Horowitz et al., J. Org. Chem., 31, 205 (1966) tritylated and mesylated A portion of this material (2.7 g, 4, 2 mmol) was dissolved in dry DMF (55ml) containing lithium azide (0.62 g, 12,7mMol) and heated for 24 hours under nitrogen at 70 ⁰ C. The reaction mixture was poured onto 400 ml of ice, the precipitate collected and purified by flash chromatography. Elution with CH 2 Cl 2 / MeOH (50: 1, v / v) afforded 1.48 g of the azido intermediate. This material was treated with 50% acetic acid (100ml) treated at 100 ⁰ C for 3 hours. Dilution with water, filtration of the resulting suspension and evaporation of the filtrate in vacuo afforded 710 mg of an oil. This oil was dissolved in 50 ml of sodium hydroxide solution and stirred at ambient temperature for 2 hours. The pH of the solution was brought to 3, the precipitate was filtered off and dried in air. 240 mg (0.7 mmol, 50%) of the title compound were obtained. Melting point = 250 ° C

 U (nm): at pH 1, Xmax = 301 (ε = 19500), amine = 230 (ε = 3,600), ACHulter = 249 ε = 12700); at pH 13 Xmax = 299, 267 (ε = 1 400.13200), Xmin = 232, 239 (ε = 1200, 7560).

H ¹ NMR (DMSO-d ₆ : 6 3.13 (s, 7H, H ₆ ), 67.32 (d, 1H-CH =, δ = 15.87 Hz), δ 6.78 (d, 1 H, = CH-COOH, J = 15.62Hz), = 6.01-5.98 (m, 1H, HV), = 5.11-5.98 (m, 1H, 5'-OH), = 4.50 ^ -4.43 (m / 1H, H3 '), = 4.13-4.08 (m, 1H, H4'), = 3.80-3.75 (m, 2H, H5 '), = 2.77-3.67 (m, 1H, H 3'), = 2.30-2.20 (m, .1H, H 2 ')

Analysis for C ₁₂ H ₁₃ H ₅ O ₆ .1.5H ₂ O: Calculated: C 41.15%, H 4.60%, N 19.99%; Found: C41.38%, H4.50%, N20.01%.

Example 71

(E) -3- [1- (3'-azide, do-2 ', 3'-dideoxy- / 3-D-erythro-pentofuranosyl) - (2,3,4-tetrahydro-2,4-dioxo) 5-pyrimidinyl] -2-propionic acid

The 5'-trityl-3'-mesyl-5-propenoate-2'-deoxyuridine was prepared according to the method of Example 69. This material was hydrogenated with 10% Pd / C in EtOH to produce the propanoate derivative. This product was then treated as described above to afford the title compound. M.p. = 118 ⁰ C to 12O ⁰ C. UV (nm): at pH 1 Xmax = 265 (ε = 9900), Xmin = 235 (ε = 3100); at pH 13 Xmax = 265 (ε = 7400), Xmin = 247 (ε = 5400)

H ¹ NMR (DMSO-dg x): δ 7.68 (s, 1H, H 6), δ 6.11-6.06 (m, 1H, H 1 '), δ 4.45- ^, 35 (m, 1H, H3 '), δ 3.85-3.80 (m, 1H, H4'), δ3.68-3.53 (m,

Analysis for C I ₂ H ₁₅ N ₅ O H 4.65 % N 21.53 %; Calculated: C 44,31 ' H 4,68 % N 21,49 %. Found: C 44,26 ' 6 ^: '/O. Vo

Example 72 Clinical Study

3'-azido-3'-deoxythymidine (AZT) was orally administered to 2 AiDS patients at a dose of 2 mg / kg every 8 hours on day 1 and on day 2 of the treatment. On days 2 and 3, patients were also given 500 mg of Probeneid (PB) every 6 hours and a single dose of AZT was given on day 3. The peak (C _ma) <and lower (C _min ) values for AZT a day (after administration of sample levy) were significantly higher than the corresponding levels on day 1, resulting in a three-fold decrease in total body clearance (Cl / F) and an increase in mean half-life (tl / 2) of AZT (0.88 to 1.73h) during PB treatment. The mean ratio of AZTGIucuronide / AZTim urine was significantly reduced from 7.3 to 2.4 after PB treatment. The main par'makokinetic parameters of AZT before and after coadministration of PB are summarized in Table I below.

Table I

Patient AUC C _max C _min T _max Cl _dead / F tl / 2

No (hVm) (/ im) (^ m) (h) (ml / min / 70kg) (h)

1. AZT / PB 10.41 6.13 0.15 0.25 829.50 1.84 Second AZT / PB 10.00 6.46 0.27 0.50 921.00 1.61 MEAN 10.21 6.30 0.21 0.38 875.25 1.73 ± SD 0.29 0.23 0.08 0.18 64.70 0.16 1. AZT 3.70 3.74. 0.00 0.50 2 333.80 0.87 Second AZT 3.04 2.51 0.00 0.31 3 027,00 0.89 MEAN 3.37 3.13 0.00 0.41 2 680.40 0.88 + 0.47 0.87 0.00 0.13 480.17 0.01

Example 73 Antiviral activity

Enhancing the Anti-Friend Leukemia Virus (FLV) Activity of 3'-Azido-3'-Deoxythymidine (AZT) by the Nucleoside Transport Inhibitors Dipyridamole, Dilazep and 6 - [(4-nitrobenzyl) thio] -9- / 3-D-ribofuranosyl) purine is shown in Table II. FG-10 cells were vaccinated on a plate one day before they were infected with FLV. One hour after infection, known concentrations of the compounds or combinations to be tested were added. The plates were incubated for 3 days, the media replaced with fresh McCoy's 5A media and incubated for a further 3 days. Concentrations of compounds to be tested / combinations giving 50% inhibition of plaque were determined as shown in Table II. Neither dipyridamole (10 / xm) nor dilazep (5μ.Μ) alone had any observed antiviral activity.

Table II

Compound / combination azidothymidine ED ₅₀ (nM)

AZT 5.

AZT + 1 μ dipyridamole 1

AZT + 5 μ dipyridamole 0.5

AZT + 10μ dipyridamole 0.2

AZT + 5 μ Dilazep 0.5 AZT + 5 / x6 - [(4-nitrobenzyl) thio] -

9- / 3-D-ribofuranosyl) purine 3

Example 74 Anti-ITP activity of 3'-azido-3'-deoxythymidine (AZT)

A patient with a platelet count of 38000mm ^"3 was a thrombozytopänische purpura (Thrombozytenzah! <100000 mm" diagnosed ³⁾ and he was treated with 5 mg / kg AZT treated for 6 weeks intravenously every 6 hours, during which time his platelet count to 140,000 mm " ³ increase.

The treatment was then changed to 5 mg / kg / 4 hours orally for 4 weeks, exposed for 4 weeks, followed by a drop in platelet count to 93,000 mm- ³ at 2 weeks and to 700,000 mm- ³ at 4 weeks. The treatment was resumed orally at 5 mg / kg / 4 hours for 5 weeks and the platelet count increased to 194,000 mm ³ . A reduction to 2.5 mg / kg / 4 hours orally resulted, in an undefined manner, in a slight decrease in platelet count but not in extensive diagnosis of ITP.

Example 75 Treatment of Kaposi's sarcoma with 3'-azitio-3'-deoxythymidine (AZT)

In this study, 9 patients diagnosed with Kaposi's sarcoma (KS) were treated with AZT and observed the following effects:

Complete healing was achieved in one patient. 3 patients showed regressions of the lesions.

In 2 patients the CS remained stable.

In 3 patients the disease continued.

The response of = 50% is comparable to that achieved with treatment with the currently preferred therapy for KS, the recombinant α-interferon.

Example 76 AZT / acyclovir combinations against HIV in vitro

Using an analogous method to that of Example 79, combinations of AZT and acyclovir (ACV) were tested for in vitro activity against HIV.

ACV alone showed low activity, a concentration of 16 / xg / ml (the highest that was studied) had less than 30% protection, while AZT at 8μ, Μ showed 100% protection by this method.

Table III shows those combinations of the two agents required to achieve 100% protection.

Table III ACV (/ xg / ml AZT (μΜ)   - O 8th 0.5 2 1 1 4 O 8th

These results indicate that ACV potentiates about three times the antiviral activity of AZT.

Example 77

AZT / interferon combinations against HIV in vitro

Peripheral blood mononuclear cells (PBMC) from a healthy HIV seronegative volunteer donor were obtained by Ficoll-Hypaque sedimentation of heparinized blood. The cells were treated with 10 μg / ml phytohemagglutinin (PHA) and in the medium RPM11640 supplemented with 20% fetal calf serum (FCS) antibiotics, 1-glutamine and 10% interleukin-2 (IL-2) (Electronucleonics, Bethesda, MD). The cells were given below χ between 4 to 6 days after exposure to PHA concentrations of 4 10 ^s cells / ml in 25 cm ³ flask, which contained 5ml medium, distributed, and exposed to the action of the means and the virus, as detailed is. The day of viral inoculation is designated as day 0. On day 4, fresh medium was added. Every 3 to 4 days thereafter, a portion of the cell suspension was removed for analysis and replaced with cell-free medium. Experiments 2 to 4 were terminated after 14 days and trials 1 and 3 after 16 days.

The viral materials were cell-free supernatants from HIV-infected H9 cells frozen in aliquots at -70 ° C. The 50% tissue culture infection dose (TCID ₅₀ ) of virus material was 10 ⁵ / ml.

Four separate experiments were performed using PBMC from different donors (Table IV). In Experiment 1, both agents were given after the cells had been exposed to the virus. Aliquots of 40 x 10 ⁶ cells were suspended in 20 ml of medium containing 10 ⁵ TCID ₅₀ virus for 1 hour, then washed 3 times and resuspended in virus-free medium. In experiments 2 and 4, the cells were incubated for 24 hours with or without recombinant a-interferon (rlFNaA), then exposed to AZT and virus. The viral inocula were 4 × 10 ³ , 10 ³ and 2 × 10 ³ TCID ₅₀ in experiments 2, ³ and 4, respectively. Concentrations of the agents were adjusted with each change in medium to maintain original concentrations.

In all experiments, two-fold dilutions of a fixed combination of rN FNaA and AZT were serially studied.

Each combination of HFNaA and AZT used in combination was also studied on its own to provide benchmarks.

Duplicate cultures for each concentration and for infected and uninfected controls were kept in stock in all experiments. In experiment 1, 3.2μ.Μ AZT and 128 units / ml (U / ml) of rlFNaA, as well as 5 double dilutions of this combination, were assayed.

In Runs 2 and 3, 0.15 μΜ AZT and 128 U / ml of rlFNaA and 3 to 4 times 2-fold dilutions of this combination were used. In experiment 4, 0.08 / nM AZT, along with 128 U / ml of rlFNaA and 2 double dilutions of this combination were used.

After approximately one week, the cultures were scored every 3 to 4 days for the presence of virus. The cells were scored for HIV antigens by indirect immunofluorescence; the supernatant fluids were evaluated for their reverse transcriptase (RT) activity, viral yield and HIV p24 antigen by radioimmunoassay.

. The multiple drug effect analysis of Chou and Talalay (Advances in Enzyme Regulation, [1984], 22, 27-55) was used to calculate combined drug effects.

Also, data were numerically evaluated by the isobologram technique, a geometric method for determining drug interactions. The concentration of AZT producing a desired (e.g., 50% inhibitory effect) effect is plotted on the horizontal axis and the concentration of rlFNaA producing the same effect on the vertical axis. A line is drawn connecting these points and the concentration of the agents in the combination producing the same effect is plotted. If this point falls below the line, the combination is considered synergistic.

In Experiment 1, all concentrations of AZT were fully inhibitory, making it impossible to assess combination effects. In experiments 2 to 4, a synergistic interaction of the two agents was consistently observed (Tables IV to IX). Synergism was evident in all measures of applied viral replication and persisted, although the effect of rlFNaA alone was negligible. Synergy calculations were performed by applying the multi-drug effect analysis to the RT data from Runs 2 to 4, the virus yield data from Runs 2 and 3 and the RIA data from Run 4. The isobologram method also showed synergism

Table IV

attempt

HIV

Ino.kulations-

method*

HIV supplement (TCID ₅₀ )

Time of addition of funds' rlFNaA AZT

A B B B

10 ⁵

4 x 10 ³ 2x10 ³

-24 h -24h -24 h

Method A: 40 x 10 ⁶ cells are suspended in 20 ml of the medium containing 10 ⁵ TCID ₅₀ HIV, suspended at 37 ° C for 1 hour, then washed and resuspended.

Method B: The indicated amount of virus was added to 2 × 10 ⁶ cells in the medium; the cells were subsequently not washed.

Table V

Try 2 - day

Effects of HFNaA and AZT on Mean Reverse Transcriptase Values (cpm / 10 ⁶ cells χ 10 ³

AZT (μΜ)

HFNaA (U / ml) 0

16

64

205

159

110

71

31

173 85

150 42

169

Table VI Trial 3 - day

Effects of HFNaA and AZT on mean RT values (cpm / 10 ⁶ cells x 10 ³ )

AZT (μΜ)

HFNaA (UAnI) 0

32

128

0.02 0.04 0.08 0.16

157

51

10

143 10

117

130

Table VII Trial 4 - day

Effects of HFNaA and AZT on mean RT values (cpm / 10 ⁶ cells χ 10 ³ ) AZT (μΜ)

HFNaA (U / ml) 0

64

0.02 0.04 0.08

32 8

4 1

Table VIII Trial 3 - day

Effects of HFNaA and AZT on virus yield (TCID _S0 / ml)

AZT (μM)

HFNaA (U / ml) 0

16

64

128

₁₀ 5.7 ₁₀ 4.8

10 ³

10 ¹ · ⁹

₁₀ 5.6

10 ¹ · ⁹

nS.1

10 ⁵

Table IX experiment 4 - day 14

Effects of rlFNaA and AZT on HIV p24 *

IFNaA (U / ml) AZT (μΜ). 0 32 64 128

0 200-300 100-200 50-100 25-30

0.02 100-200 2.2

0.04 25-30 1.0

0.08 11.2, 0.8

* HIV p24 values are expressed as ng protein / ml.

Table X

Combination indices for AZT and HFNaA calculated from RT data

Combination indices at different percentages of RT inhibition

Day in trial culture 50% 90% 95%

2 7 2.14 0.34 0.23 2 10 0.37 0.30 0.28 3 6 0.26 0.73 1.39 3 13 0.12 0.15 0.17 3 16 <0.01 0.02 0.05 4 8th 0.01 0.03 0.04 4 14 0.02 0.07 0.12

The C.I. values are determined by solving the equation for different degrees of RT inhibition. C.I. values <1 indicate synergism. The given C.I. values are obtained using the mutually non-exclusive form of the equation; Values obtained using the mutually exclusive form were always slightly lower.

Example 78 Antibacterial synergy studies in vitro

3'-azido-3'-deoxythymidine and 8 known antibacterial agents (listed in Table XI) were treated with Ν, Ν-dimethylformamide every 30 minutes. Microtiter dilutions were made using "Wellcotest broth." Prior to synergy, MIC determinations for each compound were made individually against the test organism (E. coli CN 314). Table XI shows the MIC endpoints of each agent for E. coli CN 314.

Twofold serial dilutions of the agents to be tested or of 3'-azido-3'-deoxythymidine were prepared in "flat bottom" and "transfer" microtiter plates, respectively. The plates containing the appropriate dilutions were pooled, yielding series of 192 dilutions / combination. Exclusive controls from the compound were also included. The highest concentration of each applied agent was twice its MIC (Table XI).

The test plates were inoculated with a bacterial inoculum containing approximately 5 × 10 ⁵ CFU / ml and then incubated at 27 ^° C. for 18 hours. The wells were evaluated for bacterial growth or non-growth and the MIC values determined. Fractional inhibitory concentrations (FICs) were calculated from MIC values by dividing the MIC values associated with the MIC of each individual agent, and the sum in fractions (the sum of fractional inhibitory concentrations) was calculated, a result of about 0 , 5 or below indicates synergy.

Table Xl Minimal inhibitory concentrations (MIC values) of the investigator alone

connection MIC (/ ig / ml) tobramycin 0.4 fusidic 1000 chloramphenicol 3.1 clindamycin 100 erythromycin 25 rifampicin 6.2 3'-azido-3'-deoxythymidine 1.0 trimethoprim 0,125 Sulfadimidin 32

The results of the synergy experiments are shown in Table XII.

TabelleXII Synergy studies: combinations of AZT and other antibacterial agents

Combination of optimal MIC values ^ g / ml):

Medium / AZT FIC index

Tobramycin / AZT 0.2 / 0.125 0.25 Fusidic / AZT 250 / 0.03 0.28 Chloramphenicol / AZT 1.6 / 0.06 0.31 Clindamycin / AZT 12.5 / 0.25 0,375 Erythromycin / AZT 6.2 / 0.25 0.5 Rifampicin / AZT 3.1 / 0.06 0.56 Trimethoprim / AZT 0.004 / 0.5 0.504 Sulfadimidin / AZT 0.25 / 0.125 0,375

Example 79 Anti-HIV activity of 3'-azidonucleosides in vitro

The in vitro activity of S'-azidonucleosides (drugs) was determined in two cell lines; H 9 (OKT4 ⁺ HT cell line, which allows HIV replication but is partially resistant to the cytopathic effect of HIV) and TM3 (T cell h clone, specific for tetanus toxoid, immortalized by lethally irradiated HTLV-I and selected for rapid growth and sensitivity to the cytopathic effect of HIV).

The inhibition test was performed as follows: TM3 cells were stimulated by antigen plus irradiated (4000 rad, 40Gy) fresh autologous peripheral blood mononuclear cells (PBM) and in a complete medium containing 15% (v / v) interleukin-2 (PBM). IL-2, lectin-depleted; Cellular Products, Buffalo, NY) cultured 6 days prior to testing. ATH 8 cells were used without the antigenic stimulation. After pre-exposure to 2 μg of polybrene per ml over a period of 30 minutes, the target T cells (2 × 10 ⁵ ) were pellitized, exposed to HIV for 45 minutes, resuspended in 2 ml of fresh medium and placed in culture tubes at 37 ° C incubated in 5% C0 ₂ containing moist air. Control cells were treated similarly but not exposed to the virus. The cells were continuously exposed to IL-2 and the drug. When ATH8 cells were used in this assay system, five virus particles per cell were the minimal cytopathic dose of the virus. In cell co-culture experiments, 5 × 10 ⁴ lethally irradiated (10,000 rad) HIV RF-II-producing H9 cells or uninfected H9 cells were added to 2 × 10 ⁵ target T cells. At these time points, total viable cells in the hemocytometer were counted under the microscope by the tryptan blue dye exclusion method. The results are shown in Table XIII below.

Table XIII

Connection ". ED ₅₀ (AtM)

3'-azido-2 ', 3'-dideoxycytidine 10

3'-azido-5-bromo-2 ', 3'-dideoxyuridine 5

3'-azido-5-bromo-2 ', 3'-dideoxycytidine 5 (E) -3- [1- (3'-azido-2', 3'-dideoxy-j3-D-

erythro-pentofuranosyl) -1,2,3,4-tetra-

hydro-2,4-dioxo-5-pyrimidinyl] -2-acrylic acid 100

Example 80 Antibacterial in vitro activity of 3'-azidonucleosides

Table XIV shows the in vitro antibacterial activity of 3'-azidonucleosides in the form of the minimum inhibitory concentration (MIC) against various bacterial species.

The standard used was trimethoprim (TMP) and the medium used was Wellcotest Sensitivity Test Agar plus 7% lysed horse blood.

Table XIV MIC (Mg / ml) 100 3 10 4 0.1> 5 10 6 0.1 §> 7 8th 10 default connection 10 10 0.1 > > 100 10 10 > 100 organism 1 2 0.1 > 10 0.1 ^ 0.1> 10 10 0.1 > > 100 0.1>> 10 > 100 > 100 100 10 10 > 100 0.5 E.coli 0.1> 10 10 0.1 > 10 10 10 > 100 0.1 Salmonella typhimuri around 0.1 ä> 10 0.5 Salmonella typhosa 0.1 => 0.5 Entero. aerogenes 10 Citro.freundii

The compounds used were:

1 = 3'-azido-3'-deoxy-4-thiothymidine

2 = 3'-azido-3'-deoxy-5'-O-acetyl-4-thiothymidine

3 = 3'-azido-3'-deoxy-2-deoxy-2-thiothymidine

4 = 6'-acetyl-S'-azido-S-benzoyl-S'-deoxythymidine

5 = i-fS'-O-acetyl-S'-azido ^ '' - deoxy-zS-D-erythro-pentofuranosyD-B-methyl ^ -d ^^ -triazol-i-yD ^ dHJ-pyrimidinone

6 = 1- (3'-azido-2 ', 3'-dideoxy- / 3-erythro-pentofuranosyl) -2- (benzyloxo) -5-methyl-4- (1H) -pyrimidinone

7 = 3'-azido-3'-deoxy-2-methoxythymidine

8 = 3'-azido-5-bromo-2 ', 3'-dideoxyuridine

Claims (6)

wherein C is a purine or a pyrimidine base attached at the 9- or 1-position, respectively, and pharmaceutically acceptable derivatives thereof, other than (a) compounds of formula (I) B, wherein C is an adenine, guanine, uridine, cytidine or thymine base and their 5'-mono or 5'-triphosphate esters; (b) the 5'-O-acetate, 5'-0-trityi and 5'-O- (4-methylbenzenesulfonate) derivatives of the compound of formula (I) B, wherein c is a uridine base and the 3'-azido group in dererythro configuration is present; (c) compounds of formula (I) B wherein C (i) is a B-bromo-vinyluridine or B-trifluoromethyl-uridine moiety and the 3'-azido group is in the erythroid configuration; (ii) is a uridine moiety and the 3'-azido group is in the threo configuration; and (iii) is a 5-iodo or 5-fluorouridine moiety and the 3'-azido group is in the erythro-orerthreo configuration; and 5'-O-trityl derivatives of such compounds; (d) compounds of formula (I) B, wherein C (i) is a 5-bromo-vinyluridine or cytidine radical and the 3'-azido group is in the threo configuration; (ii) is a 5-fluorocytidine moiety and the 3'-azido group is in the erythroid configuration; or (iii) is a 5-methylcytidine residue and the 3'-azido group is in the threo or erythro configuration; (e) the 5'-O-acetate esters of compounds of formula (I) B in which C is a 4-chloro 2 (1H) pyrimidinone or 4- (1H-1,2,4-triazol-1-yl) -2 (1H) pyrimidinone (optionally substituted at the 5-position by fluoro or methyl) and the 3'-azido group in the erythro configuration; (f) the 5'-0 - [(4-methoxyphenyl) -diphenylmethyl] derivative of the compound of formula (I) B, wherein C is a cytidine moiety and the 3'-azido group is in the erythroid configuration; and (g) the 5'-O-trityl derivative of the compound of formula (I) B, wherein C is an adenine residue and the 3'-azido group is in the threo configuration; characterized in that one
(A) a compound of the formula (III) (wherein A is a purine or pyrimidine base other than thymine attached at the 9 and 1 positions, respectively, and Μ is a precursor group for the 3'-azido group) or a derivative thereof, with an agent or under conditions which to convert the precursor group into the desired azido group, reacting; or (B) a compound of the formula (IVa) (wherein R 1, R ² , R ³ , R ⁴ , R f and R ⁷ , respectively the groups R ¹ , R ² , R ³ , R ⁴ , R ⁶ and R ⁷ ) and R ^{1 is} hydroxy, mercapto, amino, alkylthio, aralkoxy, alkoxy, cyano, alkylamino, wherein the alkyl groups are optionally joined to form a heterocycle are; R ^{2 is} hydrogen, acyl, alkyl, aroyl or suifonate; R ^{3 is} hydroxy, mercapto, amino, triazolyl, alkylamino, dialkylamino, wherein the acyl groups are optionally joined to form a heterocycle, aralkoxy, alkoxy or alkylthio; R ^{4 is} alkyl, substituted alkyl, halogen, perhalomethyl, hydroxy, alkoxy, cyano, nitro, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl or hydrogen; and R ⁶ and R ^{7 are the} same or different and are selected from amino, hydrogen, hydroxy, mercapto, alkylthio, alkoxy, aralkoxy, cyano or alkylamino; or precursor groups therefor, provided that at least one of R], R ² , R ³ and R ⁴ in formula (IVa) or at least one of Rg and Rg in formula (IVb) represents a precursor group) with an agent or under conditions which lead to the conversion of the precursor group (s) into the corresponding desired groups; or a compound of the formula (wherein R ¹ , R ² , R ³ , R ⁴ , R ⁶ and R ^{7 have} the same meaning as above) or a functional equivalent thereof, with a compound capable of introducing the desired ribofuranosyl ring at the 1-position of the compound of the formula (IVa) or the 9-position of the Formula (IV b) is used, implemented; or if A of the compound of formula (I) is a purine residue, a purine of formula (Vb) having a Pyrimidine core of the formula _f HO (wherein Py represents a 1-pyrimidinyl group); and subsequently, or concurrently with, one or more of the following transformations optionally carried out: (i) if a compound of formula (I) is formed, converting the compound into a pharmaceutically acceptable derivative thereof; and (ii) if a pharmaceutically acceptable derivative of a compound of formula (I) is formed, converting the derivative into the parent compound of formula (I) or into another pharmaceutically acceptable derivative of a compound of formula (I). 2. Method according to item 1 for the preparation of a S'-azido-S'-deoxypyridimidinnucleoside, or a pharmaceutically acceptable derivative thereof, characterized in that R ^{1 is} hydroxy, mercapto, C ₁ _ "alkoxy or amino; R ^{2 is} hydrogen, methyl, C ₁ _ ₂ alkanoyl or benzoyl; R ^{3 is} hydroxy, mercapto, amino or substituted amino; and
R ⁴ is hydrogen if R ³ is amino or substituted amino, and halogen, perhalomethyl, C ₁ _ ₃ alkyl, C ₂ _ ₃ alkenyl, or substituted ethenyl, if ^R3 is not amino or substituted amino. 3. The method of item 1 for the preparation of an S'-azido-S'-deoxypurinnucleosids, or a pharmaceutically acceptable derivative thereof, characterized in that R ^{6 is} amino, C ₁ -4-alkylamino, mercapto, hydroxy or Ci _4-alkoxy ; and R ^{7 is} amino, Ci _4-alkylamino or hydrogen. 4. The method according to any one of items 1 to 3, characterized in that the 3'-azido group is present in the erythro configuration. 5. The method according to any one of items 1 to 4, characterized in that the group C of formula (I) B is a guanine, adenine, thymine, uridine or cytidine derivative. 6. The method according to any one of items 1, 2, 4 and 5, characterized in that the group C is a thymine or uridine derivative and the 3'-azido group is in the threo configuration.