DD262802A5 - PROCESS FOR PREPARING A PHARMACEUTICAL FORMULATION - Google Patents

Abstract

The invention relates to a process for the preparation of a pharmaceutical formulation containing certain novel 3-azidonucleosides and pharmaceutically acceptable derivatives thereof. The new formulations contain special combinations of active ingredients. They are particularly useful in the treatment and prophylaxis of Gram-negative bacterial infections and retroviral infections, particularly AIDS.

Description

Field of application of the invention

The invention relates to a process for the preparation of a pharmaceutical formulation.

Characteristic of the known state of the art:

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 opportunities 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, because of the close similarity 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 non-toxic 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 form a subset of the RNA viruses that must first "reverse-translate" the RNA of their genome into the DNA for replication (the "back translation" or "transcription" conventionally 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 (HTLV III) as an AIDS-accompanying retrovirus (ARV) or as a lymphadenopathy-accompanying virus (LAV), however, the internationally recognized name of human immunodeficiency virus (human immunodeficiency virus 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 Kaposi'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 leading to premature wear and symptoms such 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, based on antiviral activity relative to FLV and a lack of cellular toxicity, 3'-azido-3'-dideoxythymidine "bromodeoxyuridine However, DeClerq et al. (Biochem Pharm., 1980) 29, 1849-1851) found six years later that 3'-azido-3'-dideoxythymidine had no appreciable activity against any viruses used in their experiments, including such DNA viruses as vaccinia, HSIV and varicella zoster virus (VZV).

Bacteria also pose a problem in therapy because all living organisms use nearly the same CVs as the others, so that one substance that is toxic to one is likely to be toxic to another as well. 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

With the invention, the shortcomings of the prior art are to be eliminated.

Explanation of the essence of the invention

The invention has for its object to provide a novel process for the preparation of a pharmaceutical formulation containing certain novel 3'-Azidonucleoside or pharmaceutically acceptable derivatives thereof. The invention is based on the discovery that certain 3'-azidonucleosides as described above are useful in the therapy of viral and bacterial infections, especially retroviral infections, including HIV infections and Gram-negative bacterial infections, including certain strains of Gram-negative bacteria are resistant to the commonly used antibacterial agents.

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.

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

 According to the invention, the formulation is prepared by

a) preparing a compound of formula (I) wherein B is a purine or pyrimidine base attached at the 9 or 1 position, or a pharmaceutically acceptable derivative thereof, by a process comprising:

(A) a compound of the formula (III) (wherein A is a purine or pyrimidine base attached at the 9 or 1 position, respectively, and M is a precursor group for the 3'-azido group) or a derivative thereof, with an agent or under conditions which result in the conversion of the precursor group into the desired azido group; or

(B) a compound of formula (IVa) or (IVb) (wherein R], Ri RjJ, Rf, Rf or R ⁷ , 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 linked to form a heterocycle;

R ^{2 is} hydrogen, acyl, alkyl, aryl or sulfonate;

R ^{3 is} hydroxy, mercapto, amino, triazolyl, alkylamino, dialkylamino wherein the alkyl groups are optionally joined to form a heterocycle, aralkoxy, alkoxy or alkylthio;

R ^{4 is} alkyl, substituted alkyl, halo, perhalomethyl, hydroxy, alkoxy, cyano, nitro, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl or hydrogen; and

R ^e 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, assuming; in that at least one of the radicals R ¹ , R ² , R ³ , R ⁴ in the formula (IVa) or at least one of the groups R ⁶ and R ⁷ in the formula (IVb) represents a precursor group, with an agent or under conditions which Conversion of the precursor group (s) into the corresponding desired groups lead, implemented; or

(C) a compound of formula (Va) or (Vb) (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, which reacts to introduce the desired ribofuranosyl ring at the 1-position of the compound of formula (IVa) or the 9-position of formula (IVb); or

(D) if B of the compound of formula (I) A is a purine residue, reacting a purine of formula (Vb) with a pyrimidine nucleus of formula (VI) (wherein Py represents a 1-pyrimidinyl group); and subsequently or simultaneously with one or more of the following transformations, if appropriate;

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

(ii) when a pharmaceutically-acceptable derivative of a compound of formula (I) A is formed, the derivative is converted to the parent compound of formula (I) A or to another pharmaceutically acceptable derivative of a compound of formula (I) A; and

b) the resulting compound of formula I (A) or a pharmaceutically acceptable derivative thereof interacts with at least one further pharmaceutically active ingredient and a pharmaceutically acceptable carrier therefor.

It is expedient to prepare a compound of the formula (I) A according to process A in which B is a thymidine base and the 3'-azido group is in the erythroid configuration. It is further advantageous to interact the compound of formula (I) A or a pharmaceutically acceptable derivative thereof with acyclovir (9- (2-hydroxyethoxymethyl) guanine). The compound of the formula (I) A or a pharmaceutically acceptable derivative thereof may be interacted with an interferon.

Specific bacteria against which good activity has been found are the following: Escherichia coli. Salmonella dublin. Salmonella typhosa, Salmonella typhimurium, Shigella flexneri, Citrobacter freundii, Klebsiella pneumoniae, Vibrio cholerae, Vibrio anquillarum, Enterobacter aerogenes, Pasteurella multocida, Haemophilus influenzae, Yersinia enterocolitica, Pasteurella haemolytica, Proteus mirabilis and Proteus vulgaris, the causative organisms of such conditions as Traveler's diarrhea, urinary tract infections, shigellosis, abdominal typhoid fever (typhus abdominalis) and cholera in humans, as well as animal diseases such as enteritis in newborn calves, enteritis in pigs after weaning and colisepticaemia in chickens.

The formulations 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.

The compound of the 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 optimal ratio is usually employed to ensure maximum exponentiation, even a miniscule amount of one agent will suffice 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: 500, 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 that 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 γ, while nucleoside transport inhibitors include such agents as dilazep, dipyridamole, 6 - [(4-nitrobenzoyl) thio] -9- (β-D-ribofuranosyl) purine, papaverine, mioflazine, hexobendin , Lidoflazine and their acid addition salts.

Particularly preferred types of interferon are α, β and γ, while nucleoside transport inhibitors include such agents as dilazep,.

Dipyridamole and 6 - [(4-nitrobenzoyl) thio] -9- (β-D-ribofuranosyl) purine.

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-compete for 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 No. 1523865, U.S. Patent No. 4,360,522 EP-PSs 74306, 55239 and 146516 and European Patent Applications 434393 and 434395, and in particular those compounds of the general formula (A) 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 -I 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 formula (A). The above-mentioned aralkanoyloxy and aroyloxy ester groups may be substituted, for example, by one or more halogen atoms (eg, chlorine or bromine), or amino, nitrile, or sulfamido groups, the aryl moiety of the group advantageously containing from 6 to 10 carbon atoms.

Particularly preferable 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 especially 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 is 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: benzylpyrimidine, 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. For example, sulfadimidine; rifampicin; tobramycin; fusidic; chloramphenicol; Clindamycin and erythromycin.

Accordingly, combinations in which 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 is, for example, interleukin II, suramin, phosphonoformate, HPA 23,2 ', 3'-dideoxynucleosides, for example, 2', 3'-dideoxycytidine and 2 ', 3'-dideoxyadenosine, or medicaments, such as levamisole or thymosin for increasing lymphocyte counts and / or function, 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 LvmDhozvtentransDlantationen.

Certain of the retroviral infections described above, for example 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 the formula (I) and (I) A for use in the present invention are those of the formula (II) wherein

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 wherein the alkyl 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 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 or 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. B. 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 (e.g., compounds of formula (I) wherein R ^{3 is} not an amino or alkylamino group) wherein the 3'-azido group is in either the erythro or threo 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 formula (II A), wherein R ⁶ and R ^{7 may be the} same or different and 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, for. 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 acyl groups advantageously contain alkyl or aryl groups, as described below. With respect to the above compounds of the formulas (II) and (III 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 9 n aralkoxy, 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 the radicals R ^1, R ^2, R ³ and R ⁴ are as defined below, namely R ¹ is hydroxy, mercapto, C ₁ ^ -alkoxy or amino; R ² is hydrogen, methyl, Ci_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 ethylen 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 (e.g., succinate), C 1-6 thioesters, optionally substituted aryl esters, mesylate, glucuronide or mono-, di- or tri-phosphates. Other classes of preferred compounds of the formula (II A) include those wherein R ^{6 is} amino, C 1-4 alkylamino, mercapto, hydroxy or C 1-4 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), Ci_e-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, Mercpato, CI_ ₅ alkoxy, amino, Ce-12-aralkoxy or a 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 6 -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 residue, and especially when the mentioned compound is 3'-azido-3'-deoxythymidine.

The compounds of formula (I) which are preferred for use in the treatment or prophylaxis of bacterial infections are those of formula (II) 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, but the erythro configuration is particularly preferred, and preferred pharmaceutically acceptable derivatives are the 5'-esters of simple organic acids, preferably Ci_i _a 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) A 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 Radical, and especially when the mentioned compound is 3'-azido-3'-deoxythymidine.

By "pharmaceutically acceptable derivative" is meant any pharmaceutically acceptable salt, ester, or salt of such an ester, or any other compound capable of administration to the recipient, the parent 3'-azidonucleoside, or a therapeutically active metabolite or moiety 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 (eg methoxymethyl), aralkyl (eg benzyl), aryloxyalkyl (eg phenoxymethyl), aryl (eg phenyl, optionally substituted by halogen, Ci_4-alkyl or Ci ^ -alkoxy); 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 (ζ.

Sodium), alkaline earth metal (e.g., magnesium) salts, ammonium and NXJ (where 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) which may be used in accordance with the present invention 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; succinate; pivalate; and mesylate.

It is possible to use compounds of the formula (IB) in which C is one at the 9-resp. 1-position linked purine or a pyrimidine base and pharmaceutically acceptable derivatives thereof, different from

(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 groups are present 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 radical 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 formula (I) B in which C is 4-chloro-2 (1H) pyrimidinone or 4 - (H-1,2,4-triazole-1 -yl) -2 (; 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 deuteron 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 the 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 of body weight of the recipient per day, preferably in the range of 6 to 90 mg per kg of body weight per day, and more preferably in the range of 15 to 60 mg per kg of 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 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

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 75 μΜ, preferably about 2 to 50 μΜ, more preferably about 3 to about 30 μΜ be administered should. This can be achieved, 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 according to the invention 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 further 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, orally , 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 the 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 20 mg / 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 suitable effective dosage range of the interferon is3 χ χ 10 ⁶ to 10 10 ⁶ IU per m ² of body surface area per day, preferably 4 χ 10 ⁶ 6 10 ⁶ IU χ m ² per day. The active ingredient and the interferon should be in the ratio of about 5 mg per kg of active ingredient to 3 χ 10 ⁶ IU per m ² of interferon to about 250 mg per kg per day of active ingredient to 10 χ 10 ⁶ IU per m ² per day of interferon, preferably from about 5 mg per kg per day of active ingredient to 4 x 10 ⁶ IU per m ² per day of interferon to about 100 mg per kg per day of active ingredient to 6 x 10 ⁶ IU per m ² per day Interferon be administered. Interferon is preferably administered by injection (for example, sc, im or iv), while the active ingredient is preferably administered orally or by injection, but it 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 10 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 i. The dose of active ingredient is generally in the range of about 1.5 to 15 mg per kg per day, and the appropriate effective i. The dose of the therapeutic nucleoside is generally in the range of about 5 to 30 mg per kg per day. It is preferred that the ratio for the i. v. administration of the active ingredient to the second therapeutic nucleoside should range from 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, an appropriate effective dose of active ingredient 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 agent is in the range of 2 to 1000 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, especially 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 ci'd 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 from the 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 of the present invention 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 parenterally (including subcutaneous, intramuscular, intravenous and interdermal) administration. The formulations may suitably be presented in unit dosage form and

according to any method known to those skilled in the pharmaceutical art. Such methods include the step of combining the active ingredient with the carrier containing one or more minor ingredients. In general, the formulations are prepared by uniformly and intimately bringing the active ingredient into contact with liquid carriers or finely divided solid carriers, or both, and then shaping the product if necessary.

The formulations suitable for oral administration of the present invention 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 non-aqueous 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 accessory ingredients. Compacted tablets may be prepared by compressing the active ingredient in a free-flowing form, such as a powder or granules, in a suitable machine, optionally mixed with a binder (eg, povidone, gelatin, hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants ( 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 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 medicinal drink (eg aqueous or non-aqueous 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. B. 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, e.g. 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 the formulations of this invention may contain, in addition to the ingredients particularly mentioned above, other conventional agents which are related to the type of formulation in question, for example those suitable for oral administration, such further agents as sweeteners, Thickener 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], T.A. 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. Chem., 43 [15], [1978] 3044-3048; KAWatanabe et al., J. Org. Chem., 45, 3274 [1980]. and RPGlinski et al., (J.Chem.Soc.Chem.Commun., 915 [1970]); these references, or s.if, the use of procedures analogous to those described therein, are expressly set forth herein Referenced.

EXAMPLES

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

250 26 9 12 3 300

mg / tablet 250

26 9

12 3300

mg / tablet Formulation A (a) active ingredient 250 (b) lactose B.P. 210 (c) povidone B.P. 15 (d) Sodium starch glycolate 20 (e) magnesium stearate 5 500 Formulation B mg / tablet (a) Active ingredient 250 (b) lactose 150 - (c) Avicel PH101 60 (d) Povidone B.P. 15 (e) magnesium starch glycolate 20 (f) magnesium stearate 5 500 Formulation C mg / tablet Active ingredient 100 lactose 200 Strength 50 povidone 5 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

mg / capsule

Active ingredient 250

Pregelatinized Starch NF15 J50

400 formulation E

mg / capsule

Active ingredient 250

Lactose 150

Avicel 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 4000 BP 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 100

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 q. s. to the pH 0.200 g Formulation A q. s. to the pH 4.0 to 7.0 Active ingredient q. s. to 4.0 to 7.0 Hydrochloric acid solution, 0.1 molar 10ml Sodium hydroxide solution, 0.1 molar Sterile water

The active ingredient was dissolved in the bulk of the water dissolved (35 ° to 40 ⁰ C) and the pH adjusted to a value depending on between 4.0 and 7.0 with the hydrochloric acid or sodium hydroxide. 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 pH 7-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 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

Weight (g)

Active ingredient 0.2500

Sorbitol solution 1.5000

Glycerin 2.0000

Sodium benzoate. 0.0050

Flavoring, peach 17.42.3169 0.0125 ml

Purified water q.s.to 5,0000 ml

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.

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 (63 μηη) * 250

Hard Grease, BP (Witepsol H15 Dynamite Nobel) 1770

2020

One fifth of the amount of Witepsol H15 is melted in a pan with a steam jacket at a maximum of 45 ° C. The active ingredient is sieved through a 200 μπη 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 is added to the suspension and stirred to ensure a homogenous mix. the entire suspension is passed sieved through a 250 ^ m stainless steel screen and allowed to cool with continuous stirring to 40 ⁰ C. at a temperature of 38 ° C to 40 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 pessaries mg / pessary 250 Active ingredient (63 μπη) 380 Anhydrous dextrose 363 potato starch 7 magnesium stearate 1000

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 nucleoside as indicated.

Example 8

injection

Formulation A Weight (mg)

Acyclovir 400

Active ingredient 200

1 molar NaOH as needed

Sterile water to 10 ml

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

Formulation B

Weight (mg)

2-Amino-9- (2-hydroxyethoxymethyl) purine 400 Active Ingredient 200

1 molar NaOH as needed

Sterile water to 10 ml

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.

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

Lactose 133

Sodium starch glycolate 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 3 Mega units

Active ingredient 200 mg

Sterile buffer, pH 1 to 50ml

Dissolve the inferon 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

tablet

Nucleoside transport inhibitor Active ingredient Lactose

Strength

Polyvinylpyrrolidinone magnesium stearate

total weight

Weight (mg)

300 200 105

20

10 685

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

Kapsei

Nucleoside transport inhibitor Active ingredient Lactose

Sodium starch glycolate Polyvinylopyrrolidinone Magnesium stearate

Weight (mg) 300 100 100 10 10

Total weight 523

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

Nucleoside Transport Inhibitor Active Ingredient I Glycerin

Cetostearyl Alcohol Sodium Lauryl Sulfate White Soft Paraffin Liquid Paraffin Chlorocresol Purified Water

on

Weight (g) 7.5 5.00 2.00 6.75 0.75

12,50 5,00 0,10

100.00

The active compounds are dissolved in a mixture of purified water and glycerol and heated to 7O ⁰ C. The remaining ingredients are heated together to 70 ° C. The two parts are combined and emulsified. The mixture is cooled and filled into containers.

Example 15 Intravenous injections

(1) Active ingredient nucleoside transport inhibitor glycerin sodium hydroxide solution q. s. on water for injections

Quantity (mg) 200 300 200 pH 7.0-7.5 10ml

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 q. s. on water for injections

Quantity (mg) 100 150 125 pH 8.0-9.0 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.

The following Examples 16 and 18 describe the preparation of formulations containing 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

tablet

Active ingredient glucuronidation / renal excretion inhibitor lactose

Strength

Polyvinylpyrrolidinone magnesium stearate

Weight (mg)

100 200 105

10 total weight 485

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.

Example 17

capsule

Active ingredient glucuronidation / renal excretion inhibitor lactose

Sodium starch glycolate Polyvinylpyrrolidinone Magnesium stearate

total weight

Weight (mg)

100 100 100

323

The active compounds are mixed with the lactose and the sodium starch glycolate 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

(1) Active ingredient glucuronidation / renal excretion inhibitor glycerin sodium hydroxide solution q. s. on water for injections

Quantity (mg)

200

300

200

pH 7.2-7.5

10ml

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.

(2) Active ingredient glucuronidation / renal excretion inhibitor mannitol sodium hydroxide solution q. s. on water for injections

Quantity (mg)

100

150

125

pH 8.0-9.0

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 Granules for a medicament in the feed

Weight (g)

Active ingredient 6.0

Povidone <1.0

Lactose 93.0

Aqueous Al koholm mixq. s.

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) thio] thymidine

(a) The 5'-hydroxyl group of 3'-azido-3'-deoxythymidine (3.0 g, 11.2 mmol) was prepared by adding methanesulfonyl chloride (2.7 mL) to a solution of the 3'-azido-3'-deoxythymidine in dry Pyridine (20 ml) mesylated.

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 oil bath of 8O ⁰ C 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 flash chromatographed on silica gel eluting with CHCl ₃ : MeOH (9: 1 v / v). The product was obtained in low yield. Melting point = 184 ⁰ 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 to 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 CHCl ₃ : MeOH (95: 5 v / v.) It was necessary to chromatograph a second time on silica gel, eluting with CHCl ₃ : MeOH (98: 1 ₎ . .. 2 vol./vol) Final purification was achieved by reverse phase chromatography on C _8, eluted with water: methanol (3: 7). The yield was 2.5%.

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

S'-azido-S'-deoxy-6'-O-acetyM-d1-A-triazo-D-thymidine (Lin et al., J. Med. Chem. 26, 1691 [1983]) (1.41 g, 3 , 9 mmol) was dissolved in 100 ml of acetone and 30 ml of water and then treated with 0.39 g of 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 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 recrystallization from i-PrOH, gave a yield of 0.64 g (1.9 mmol, 48.7%). Melting point = 75 ° C to 78 ⁰ C.

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

S'-azido-S'-deoxy-O'-O-acetyl-thiothymidine (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 Stirred for 18 hours. The solvent was removed in vacuo and the residue was applied to a silica gel column, followed by elution with CHCl ₃ ZEtOAc (3: 1 v / v). The appropriate fractions were combined and the solvent removed in vacuo. A yellow oil was obtained which when dissolved in Et ₂ O yielded crystals upon concentration: 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 Ν, Ν-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 EtOAc / CHCl 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 hour reaction sequence starting 2,3'-O-anhydro-5'-tritylmidine (JJ Fox, J. Org. Chem., 28.939 [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 to 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-β-D-lyxofuranosyl) -2-thiothymine was obtained. The UV maxima at pH 1 of 277nm and at pH 11 of 241nm indicated the formation of a 2-thiothymidine. The 3'-hydroxyl group of the thiothymidine derivative was mesylated 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-β -D-lyxofuranosyl) -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-β-D-lyxofuranosyl) -2-thiothymine (0.72 g, 1, 2 mmol). 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) was purified 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 heating at reflux of 3'-azido-3'-deoxy-5'-mesylthymidine (2.6 g, 7.5 mmol) in dry ethanol (25 ml) prepared two equivalents of potassium carbonate (1.08 g, 7.5 mmol) 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 ⁰ C to 100 ⁰ C.

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

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

Example 30 S'-azido ^ 'S'-dideoxy-S-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 (Skaricand 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 ⁰ C to 108 ⁰ C.

Example 31 3'-Azido-5-chloro-2 ', 3'-dideoxyuridine

3'-azido-2 ', 3'-dideoxyuridine (0.25g, 1mmol) was dissolved in 2ml dry dimethylacetamide (DMAC), cooled to ⁰ ° C and 2ml added to a 0.5 molar HCl in DMAC. 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 (3 x 3 mL) 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 170 ⁰ C. UV (nm): at pH 1 \ max = 276 (ε = 7400), amine = 239 ((ε = 500);

at pH 13Amax = 274 (ε = 6400), \ min = 249 (ε = 3800).

H ¹ NMR (DMSO-d _e ): 58.29 (s, 1 H, H 6), 6.04 (t, 1 H, H V, J = 5.5Hz), 5.49-5.29 (m , 1H, 5'-OH), 4.44-4.20 (m, 1H, H3 '), 3.88-3.71 (m, 1H, H4'), 3.71-3.53 (m, 2H, H 5 '), 2.63-2.31 (m, 2H, H2'); Analysis for C ₉ H ₁₀ N ₅ O ₄ Cl:

Calculated: C37.58%, H3.50%, N24.35%, 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) and then bromination of the 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 ⁰ 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 ° C to 130 ° C.

Example 34 3'-azido-2 ', 3'-dideoxy-5-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 (5.0 g, 16.9 mmol) was prepared by adding triphenylmethyl chloride (5.65 g, 20.3 mmol) to the starting material in a suspension of dichloromethane (1.4 liters ml), pyridine (70 ml) and 3A molecular sieves (55 g). 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 (51 ml) 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-β-D-ribofuranosyl) -5-trifluoromethyluracil, was prepared by dissolving in nitromethane (80 ml) and adding zinc bromide (4.45 g ), dissolved in nitromethane (80 ml) 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 treatment of 1- (3'-chloro-2'-deoxy-β-D-ribofuranosyl) -5-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 for 4 hours at 90 ⁰ C. 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. DieElution on silica gel with CH ₂ CI _2: Me0H (97:. 3 vol./vol) yielded 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%. Mp = 174.5 ° C to 176.5 ⁰ C.

Example 36 S'-azido-a''-dideoxy-S-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 the 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 overnight at room temperature. 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 deblocking of the 5'-hydroxyl position was carried out by heating in 80% acetic acid on a steam bath over 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 analogs by the exact same procedure used to prepare the erythro isomer (TAKrenitsky et al., J. Chem Med. Chem., 26, 891 [1983]). The yield was 0.021g; 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 Ne-octanoyladenine (2.0 g, 7.7 mmol) and 3'-azido-3'-deoxythymidine ( 1.13g, 4.2 mmol) according to the procedure described by M.lmazawa and F. Eckstein (J. Org. Chem., 43, 3044 [1978]). M.p. = 120 ⁰ C to 122 ° C.

UVpH Umax 258nm \ min 230nm pH 13Amax 260nmAmin 229nm

Analysis for Ci ₀ H ₁₂ N ₈ O ₂ :

Calculated: C43.48%, H4.38%, N40.56%;

Found: C43.28%, H4.45%, N40.38%.

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

9- (3'-azido-2 ', 3'-dideoxy-β-D-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) according to the method of M.lmazawa and F. Eckstein, J. Org. Chem., 43, 3044 (1978). Melting point = 184 ⁰ C to 185 ⁰ C.

UVpH 1 \ max257nm \ min230nm pH 13 \ max 260nm \ min 228nm

Analysis for C ₁₀ H ₁₂ N ₈ O ₂ :

Calculated: C43.48%, H4.38%, N40.56%;

Found: C43.33%, H4.45%, N40.41%.

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-d _6):. 58.04 to 7.50 (m, 6H, 3N-benzoyl and 6H), 56.12 (dd, 1 H, J _{r, 2a} = 5.6 Hz, J _r , _2b = 6.7Hz, 1Ή), 54.55-3.96 (m, 4H, 3Ή, 4Ή, 5H '), 52.62-2.38 (m, 2H, 2'H), 52, 07 (s, 3H, 5'-acetyl CH ₃ ), 51.90 (d, 3H, J ₅ , e = 1.0Hz, 5CH ₃ ).

Analysis for Ci ₉ 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 triphenylmethyl group 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 determined to be exactly that same method as indicated for the bromination of erythro-3'-azido-2 ', 3'-dideoxyuridine.

UVpH Umax 280, = 9400, amine 244, = 2600 pH13Amax276 = 6700, amine 251 = 3700

H ¹ NMR (DMSO-d ₆ ): 511.86 (s, 1H, 3-NH), 58.02 (s, 1H, 6H), 55.99 (dd, 1H, J _r , _2a · = 3.0Hz, Jr, 2b = 7.5Hz, TH), 55.1 (t, _1H , J _{5, CHa} 5'OH = 5.4Hz, 5'OH), 54.49 (m, 1 H, 3'H), 54.05 (m, 1H, 4'H), 53.71 (m, 2H, 5'H), 52.72 (m, 1H, 2 b '), 52, 18 (m, 1H, 2a ').

Analysis for C ₉ Hi ₀ BrN ₅ O ₄ .0.25 H ₂ O. 0.1 C ₂ H ₄ O ₂ : Calculated: C 32.25%, H 3 .21%, N 20 .44%, Br 23.32%; Found: C32.17%, H3.21%, N20.33%, Br23.19%.

Example 42

1- (3'-azido-2 ', 3'-dideoxy-β-D-erythro-pentofuranosyl) -2- (benzyloxo) -5-methyl-4- (1H) -pyrimidinone Sodium (0.4 g , 17.4 mmol, 2.6 equiv. With dry benzyl alcohol (10 mL) for 1 hour at room temperature, 2,5'-O-anhydro-3'-azido-3'-deoxythymidine (1.65 g, 6.6 mmol) 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) The ethyl acetate was removed in vacuo over MgSO ₄ , 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 product-containing fractions were collected and the solvents were dissolved in Vacuum to give an oil and the oil crystallized after capping with ethyl ether, mp = 125X to 126.5 ° C.

UVpH 1 unstable

pH 13Amax 256, = 10700, Xmin 240, = 8900.

H ¹ NMR (DMSO-d ₆ ): 57.8 (d, 1 H, J ₆₅ = 1.2Hz, 6Hz), 57.49-7.38 (m, 5H, 2-phenyl), 56.08 ( dd, 1 H, J _r , _2a = 5.0Hz, J _{1> 2b} = 7.0Hz, TH), 55.37 (s, 2H, 2CH ₂ ), 55.25 (t, 1H, J _{5, C} h ₂ 5OH = 5-4Hz, 5'OH), 54.36-4.32 (m, 1H, 3'H), 53.85-3.81 (m, 1H, 4'H ), 53.7-3.58 (m, 2H, 5'H), 52.53-2.34 (m, 2H, 2'H), δ 1.82 (d, 3H, J _6-6 = 1.0Hz, 5CH ₃ ).

Analysis for Ci ₇ Hi ₉ N ₅ O ₄ :

Calculated: C57.14%, H5.36%, N 19.60%;

Found: C57.02%, H5.43%, N 19.53%.

Example 43 1- (3'-azido-2 ', 3'-dideoxy-β-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%). Melting point = 12O ⁰ C to 122 ⁰ C.

UV (nm): at pH 1 Amax = 260 (ε = 9300), amine = 237

(ε = 5500), λ shoulder = 221 (ε = 7500); beipH13Amax = 256 (ε = 10000), amine = 240 (ε = 7700).

H ¹ NMR (DMSO-d ₆ ): 57.58 (s, 1H, H6), 56.0 (dd, 1H, H1 ', J = 2.9.4, 56Hz), 55.06 (t , 1 H, 5ΌΗ, J = 4.91 Hz), 54.51-4.47 (m, 1H, H3 '), 54.34 (q, 2H ₁ -OCH ₂ -, J = 7.14Hz) , 54.10-4.05 (m, 1H, H4 '), 53.73 (t, 2H, H5', J = 5.62Hz), 52.82-2.73 (m, 1H, H2 'b) 52.21 to 2.14 (m, 1H, H2'a), 51.82 (s, 3H, 5CH _3), 51.31 (t, 3H, -CH ₂ -CH _3, J = 6.65Hz).

Analysis for C ₁₂ Hi ₇ N ₅ O ₄ :

Calculated: C48.81%, H5.80%, N23.72%;

Found: C48.59%, H5.86%, N23.64%.

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, 22 mmol) (prepared by the method of W.Sung, Nucleic Acids Research [1981] 9,6139) was dissolved in 10 μl 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 of 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 to give 0.18 g (0.62 mmol, 28%) 65 ° to 67 ° C.

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

H ¹ NMR (DMSO-d ₆ ): δ 7.63 (s, 1 H, H 1 ', J = 2.93,4.89 Hz), δ 5.06 (s, 1 H, 5ΌΗ), δ 4, 50-4.46 (m, 1H, H3 '), δ 4.10-4.04 (m, 1H, H4'), δ 3.73 (d, 2H, H5 ', J = 5.61 Hz), δ 2.78 × 2.68 (m, 1H, H2'b), δ 2.20-2.14 (m, 1H, H2'a), δ 2.00 (s, 3H, 5CH ₃ ).

Analysis for C ₁₀ H ₁₃ N ₅ O ₃ S 0.1C 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%, H 4.73%, N 23.88%, S 10.91%.

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

3'-azido-2 ', 3'-dideoxyuridine was acetylated according to the method of Visser (Syntetic Procedures in Nucleic Acid Chemistry, Vol. 1, page 410) and brominated to give 6'-acetyl-S'-azido-S bromo ^ '^' - 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. Treatment at room temperature with methanol saturated with ammonia at 0 ° C for 18 hours afforded, after recrystallization from ethyl acetate and filtration of the crystals obtained, 4-amino-3'-azido-5-bromo-2 ', 3'-Dideoxyuridine in a yield of 173 mg (0.5 mmol, 6.3%). Melting point = 162 ° to 165 ° C (dec.).

UV (nm): at pH Umax = 300.215 (ε = 10700, 12100), amine = 253 (ε = 1500); at pH 13Amax = 288 (ε = 7,300) amine = 260 (ε = 3,900).

H ¹ NMR (DMSO-d ₆ ): δ 8.3 (s, 1H, H ₆ ), δ 7.85 (broad s, 1H, 4-NH ₂ ), δ 7.05 (broad s, 1H, 4- NH ₂ ), δ 6.0 (t, 1H, H1 ', J = 5.94 Hz), δ 5.33 (t, 1H, 5'-OH, J = 5.01 Hz), δ 4.4- 4.3 (m, 1H, H3 '), δ 3.9-3.5 (m, 3H, H4', H5 '), δ 2.31 (t, 2H, H2', J = 6.27 Hz ).

Analysis for C ₉ H ₁₁ N ₆ O ₃ Br:

Calculated: C 32.64%, H 3.35%, N 25.38%, Br 24.13%;

Found: C 32.52%, H 3.41%, N 25.32%, Br 24.04%.

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

5-Bromovinyl-2'-deoxyuridine (BVDU) was synthesized using the method of Jones et al. , Tetrahedron Letters, 45, 4415 (1979) with similar yields. 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 carbonate 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 / dimethyl formamide at 13O ⁰ 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): beipHIA max = 292.247 min = 270.238; beipH13Amax = 253, amine = 238, Ash = 284.

H ¹ NMR (DMSO-d _e ): 08.06 (s, 1 H, H ₆ ), 07.25 (d, 1 H, -C = CHBr, J = 13.4Hz), 6.85 (d, 1 H, = CHBr, J = 13.7Hz), 06.07 (t, 1 H, H ₁ -J = 6.3Hz), δ 5.30 (broad s, 1 H, 5'-OH), δ 4, 42 (q, 1 H, H _3. , J = 6.3, 6.1 Hz), 53.86-3.82 (m, 1 H, H ₄ -), 53.68-3.59 (m , 2H, H ₅ ), 52.47-2.32 (m, 2H, H ₂ ).

Analysis for C ₁₁ H ₁₂ N ₅ O ₄ Br:

Calculated: C36.89%, H3.38%, N19.55%;

Found: C36.86%, H3.41%, N19.51%.

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

H ¹ NMR (DMSO-cle): δ 11.89 (s, 1H, NH), 57.94 (s, 1H, H ₆ ), 56.00 (dd, 1H, H ₁ -, J = 2,93,4,64Hz), 55.1 (t, 1H, 5'OH, J = 5.1 Hz), 54.50 to 4.46 (m, 1 H, _H3), 54.08 -4.03 (m, 1H, H ₄ -), 53.71 (t, 2H, H ₆ -, J = 5.32Hz), 52.77-2.67 (m, 1H, H ₂ . b), 52,23-2,16 (m, 1 H, H _z a). Analysis for C ₉ H ₁₀ N ₅ O ₄ Cl. 0.1 H ₂ O. 0.1 C ₄ H ₈ O ₂ : Calculated C 37.85%, H 3.72%, N 23.48%, Cl 11.89 %, Found: C37.94%, H3.91%, N23.23%, CI11.86%.

Example 48

1- (3'-azido-2 ', 3'-dideoxy-β-D-erythro-pentofuranosyl) -5-methyl-2- (pentyloxo) -4- (1H) -pyrimidinone 1- (3'-azido -2 ', 3'-dideoxy-.beta.-D-ergothipentofuranosyl) -5-methyl-2- (pentyloxo) -4- (1H) -pyrimidinone was obtained by the method which was used to prepare 1- (1) 3'-azido-2 ', 3'-dideoxy-β-D-erythro-pentofuranosyl) -2- (benzyloxo) -5-methyl-4- (1 H) -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 1Amax258nm = 9400, amine 232 nm = 5900 pH13Amax256nm = 10600, amine 239nm = 8200

NMR was performed in DMSO-d ₆ .

NMR: 57.81 (s, 1 H, 6H), 56.04 (t, 1 H, J _{r, r} = 6.7Hz, TH),

55.28 (t, 1 H, J _5, CH _2, 5 OH = 5.1 Hz, 5ΌΗ)

54.43-4.37 (m, 1H, 3'H), 54.27 (t, 2H, J ₂ _o, _1C H ₂ 2CH ₂ ) = 6.6Hz, 2-0 (CHj) ₁ ), 53 , 88-3.84 (m, 1H, 4'H), 53.70-3.50 (m, 2H, 5'CH ₂ ), δ 2.50-2.34 (m, 2H, 2'H ), 51.80 (s, 3H, 5CH _3), 51.72 to 1.68 (m, 2H, 2-O (CH _{2) 2),} δ 1.38 to 1.30

(m, 4H, 2-0 (CH ₂ J ₃ and ₄ ), 50.92-0.87 (m, 3H, 2-0 (CH ₃ J ₅ )

Analysis for C ₁₅ H ₂₃ N ₅ O ₄ · ¹ AH ₂ O: Calculated: C52,70%, H 6.93%, N 20.48%; Found: C52.63%, H6.92%, N20.48%.

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'-mesylthymidine 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 ° C to 88 ° C.

UVpH 1Amax259nm = 21,200, amine 230nm = 6400 pH13Amax265nm = 9600, amine 248nm = 8,000.

H ¹ NMR (DMSO-d ₆ ): 58.00-7.58 (m, 6H, 6H and phenyl), 56.15 (t, 1 H, J _r , _2. = 6.1 Hz, TH), 54.55-4.47 (m, 3H, 3Ή and 5'CH ₂ ), 54.12 ^ 1.10 (m, 1H, 4'H), 53.28 (s, 3H, 5'-SO ₂ CH ₃ ), 52.64-2.4 (m, 2H, 2'H), 51.89 (d, 3H, J ₅ , ₆ = 1.3Hz, 5CH ₃ ).

Analysis for C ₁₈ Hi ₉ N ₅ OyS:

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 / MeOH (v / v), followed by combining and evaporation of the appropriate fractions afforded threo-S'-azido. ''S'dideoxy-6-iodo-S'. -trityluridin. The 5'-hydroxyl was deblocked with a saturated zinc bromide / nitromethane solution at 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 13Amax = 277, amine = 252.

H ¹ NMR (DMSO-d ₆ ): 58.37 (s, 1H, H6), 56.00 (t, 1H, H1 ', J = 6.17Hz), 55.4-5.3 (m, 1H , 5'OH), 54.4-4.3 (m, 1H, H3 '), 53.9-3.8 (m, 1H, H4'), δ3.7-3.6 (m, 2H , H5 ').

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

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

Found: C30.93%, H3.09%, N16.61%, J30.94%.

Example 51

1- (5'-0-Acetyl-3'-azido-2 ', 3'-dideoxy-.beta.-D-erythro-pentofuranosyl) -5-methyl-4- (1,2,4-triazol-1-yl ) -2 (1H) -pyrimidinone 5'-Acetyl-3'-azido-3'-deoxythymidine was treated with 5 equivalents of 1,2,4-triazole and 2 equivalents of 4-chlorophenyldichlorophosphate in dry pyridine at room temperature for a period of 10 days implemented. 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 the amount of 2.7g (7.5 mmol, 60%). Melting point = 143 ° C to 145 ⁰ C.

UV (nm): at pH Umax = 324.245.215 (ε = 9300, 10000, 20500),

Amine = 282, 233 (ε = 2100, 8200); at pH 13Amax = 276 (ε6000), amine = 242 (ε = 2000).

H ¹ NMR (DMSO-d ₆ ): 59.34, 8.40 (2s, 2H, triazolyl), 58.23 (s, 1H, H6), 56.12 (t, 1H, H V, J = 6.16Hz), 54.48-4.17 (m, 4H, H3 ', H4', H5 '), 52.35 (s, 3H, 5'-acetyl), 52.07 (s, 3H, 5CH ₃ ).

Analysis for CuHi ₆ N ₈ O ₄ :

Calculated: C46.67%, H4.48%, N31.10%.

Found: C46.58%, H4.51%, N31.02%.

Example 52

1- (3'-azido-2 ', 3'-dideoxy-β-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 ⁰ C to 159 ⁰ C. >.

UV (nm): at pH Amax = 299.224 (ε = 14900.7900),

Amine = 254 (ε = 2500);

at pH 13Amax = 287 (ε = 14000), amine = 243 (ε = 6800).

H ¹ NMR (DMSO-d ₆ ): 57.63 (s, 1 H, H ₆ ), 56.06 (t, 1 H, H 1 ', J = 6.53 Hz), 55.22 (t, 1 H, 5'OH, J = 5.2Hz), 54.37-4.34 (m, 1H, H3 '), 53.83-3.80 (m, 1H, H4'), 53.65-3 60 (m, 2H, H5 '), 53.06 (s, 6H, N _(CH3 J _2), 52.29 to 2.23 (m, 2H, H2'), 52.11 (s, 3H , 5-CH ₃ ).

Analysis for Ci ₂ H ₁₈ N ₆ O ₃ :

Calculated: C48.97%, H6.16%, N28.56%;

Found: C49.06%, H6.20%, N 28.50%.

Example 53

1- (3'-azido-2 ', 3'-dideoxy-β-D-erythro-pentofuranosyl) -5-methyl-2- (isopentylxo) -4- (1H) -pyrimidinone The title compound was added by an analogous procedure used for preparing 1- (3'-azido-2 ', 3'-dideoxy-β-D-erythropentofuranosyl) -2- (benzyloxo) -5-methyl-4- (1H) -pyrimidinone (Example 42) receive.

UV (nm): at pH 1Amax258 = 9000, amine 237 = 6000 pH13Amax255 = 11000, amine 239 = 8000.

H ¹ NMR (DMSO-d ₆ ): 57.79 (s, 1H, 6H), 56.02 (t, 1H, J ₁ -, _2. = 6.3Hz, TH), 55.23 (t, 1 H, J _50h , s'h = 5.2Hz, 5ΌΗ), 54.4-4.3 (m, 3'H and 2-0-CH ₂ -CH ₂ CH (CH ₃ I ₂ ), 53 , 9-3.8 (m, TH, 4'H), 53.7-3.5 (m, 2H, 5Ή), 52.5-2.3 (m, 2'H), δ 1.78 (s, 3H, 5CH ₃ ), δ 1.7-1.5 (m, 3H, 2-OCH ₂ -CH ₂ -CH (CH ₃ J ₂ ), 50.92 and 0.89 (d, 6H, J = 6.3Hz, 2-O (CH ₂ ) ₂ CH (CH ₃ ) ₂ ).

Analysis for Ci ₅ H ₂₃ N ₅ O ₄ :

Calculated: C53.40%, H6.87%, N20.76%;

Found: C53.14%, H6.92%, N20.62%.

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) was added 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): beipH 1Amax273te = 9000), amine 251 (ε = 6200); at pH 13Amax 266 (ε = 8800), amine 247 (ε = 7000).

H ¹ NMR (DMSO-d _e ): δ 1.68 (s, 3H, 5-CH ₃ ) δ 2.47 (s, 3-COCH ₃ ), 5 2.3-2.5 (m, 2 ') H), 5 4.1-4.2 and 4.4-4.7 (m, 4H, 3Ή, 4Ή and 5'H), δ 6.1 (t, 1 H, Jv, _2. = 6, 3Hz , TH), δ 7.5-8.0 (m, 5H, 6H and phenyl).

Analysis for Ci ₉ H ₁₈ N ₆ O ₆ .0 6CHCl 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) together in dry DMSO for 2 hours

heated to 97 ° C under nitrogen. The solvents were removed in vacuo and the residual oil applied to a silica gel column followed by elution with CHCl ₃ ZEtOAc (1: 1, v / v). The appropriate fractions were pooled, combined to give, after evaporation, 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 = 16O ⁰ C to 162 "C.

UV (nm): at pH 1 Amax = 276.215 (ε = 13500, 11 200),

Amine = 238 (ε = 1700); at pH 13 Amax = 276 (ε = 10100), amine = 240 (ε = 3400).

H ¹ NMR (DMSO-dfs): δ 8.81 (s, 1 H, H 6), δ 6.00 (t, 1 H, H V, J = 5.94 Hz), δ 5.3-5.21 (m, 2H, 5'OH, 3ΌΗ), δ 4.28-4.15 (m, 1H, H3 '), δ 3.82-3.77 (m, 1H, H4'), δ 3 , 7-3.5 (m, 2H, H5 '), δ 2.18 (t, 2H, H2', J = 5.75Hz).

Analysis for Ci ₀ H ₁₁ N ₃ O ₅ 0.25H ₂ O: Calculated: C 46.61%, H 4.50%, N 16.31%; Found: C 46.67%, H 4.71%, N 16.54%.

Example 56 1- {3'-azido-2 ', 3'-dideoxy-β-D-threo-pentofuranosyl) -5-methyl-2-pentyloxy-4- (1H) -pyrimidinone 2,5'-O-anhydro -1- (3'-azido-2 ', 3'-dideoxy-β-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 cherry stalls of the title compound. Melting point = 110 ° C to 111 ⁰ C.

UV (nm): at pH 1 Amax = 255 (ε = 10200),

Amine = (ε = 2600), Ash = 222 (ε = 9000); 13Amax = 251.226 (ε = 11600.9600), amine = 238 (ε = 8600).

H ¹ NMR (DMSO-d ₆ ): δ 7.58 (s, 1 H, H ₆ ), δ 5.98 (dd, 1 H, HV, J = 2.98, 8.88 Hz), δ 5.07 (t, 1H, 5'OH, J = 5.42Hz), δ 4.5-4.47 (m, 1H, H3 '), δ 4.31-4.25 (m, 2H, -OCHH δ 4,1-4,06 (m, 1 H, Η4 '), δ 3,73 (t, 2H, H5', J = 5,62Ηζ), δ 2,82-2,72 (m, 1 H, H2 '), δ 2.2 to 2.14 (m, 1 H, H Z), δ 1.82 (s, 3H, 5-CH _3), δ 1.75-1.65 (m, 2H, pentyl), δ 1.4-1.3 (m, 4H, pentyl), δ 0.92-0.87 (m, 3H, pentyl).

Analysis for C ₁₅ H ₂₃ N ₄ O ₄ :

Calculated: C 53.40%, H 6.87%, N 20.76%; Found: C 53.31%, H 6.90%, N 20.74%.

Example 57

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

UV (nm): beipH 1 Amax = 266 (ε = 9100), amine = 235 (ε = 3000); 13Amax = 256 (8 = 11500), amine = 240 (ε = 9600).

H ¹ NMR (DMSO-d ₆ ): δ 7.6 (s, 1 H, H ₆ ), δ 7.49-7.37 (m, 5H, phenyl), δ 6.01 (dd, 1 H, HT , J = 2.52 x 5 OHz), δ 5.35 (s, 2H, benzyl), δ 5.1-5.0 (m, 1 H, 5'OH), δ 4.5-4.4 (m, 1H, H3 '), δ 4.1 - *, 0 (m, 1H, H4'), δ 3.77-3.67 (m, 2H, H5 '), δ 2.85- 2.70 (m, 1 H, H2 '), δ 2.25-2.15 (m, 1 H, H2'), δ 1.83 (s, 3 H, 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: C 56.51%, 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 O, 5N-sodium methoxide in methanol for 24 hours at room temperature. The solution was neutralized to pH 7 with a sulfonic acid resin (DOW 50W, 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 (nm): at pH Umax = 279 (ε = 7500), amine = 239 (ε = 1700);

at pH 13Amax = 279 (ε = 6400), amine = 243 (ε = 1300).

H ¹ NMR (DMSO-d ₆ ): δ 8.02 (s, 1 H, H ₆ ), δ 6.08 (t, H, H V, J = 5.86 Hz), δ 5.29 (t, 1 H, 5ΌΗ, J = 5.48Hz), δ 4.41-433 (m, 1H, H3 '), δ 3.9-3.86 (m, 1H, H4'), δ 3.86 ( s, 3 H, 4-OCH ₃ ), δ 3.75-3.58 (m, 2H, H 5 '), δ 2.4-2.3 (m, 2H, H 2'), δ 1, 89 (s, 3 H, 5-CH _3).

Calculated: C 46.97%, H 5.38%, N 24.90%; Found: C 47.06%, H 5.40%, N 24.86%.

Example 59 1- (3'-azido-2 ', 3'-dideoxy-β-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 ° C to 171 ⁰ C.

UV (nm): at pH Umax = 295, 233 (ε = 15700, 9500),

Amine = 253 (ε = 2600); at pH 13Amax = 285 (ε = 15300), amine = 241 (ε = 7900).

H ¹ NMR (DMSO-d ₆ ): δ 7.57 (s, 1 H, H ₆ ), δ 6.02 (t, 1 H, H 1 ', J = 6.5 Hz), δ 5.22 (t, 1 H, 5'0H, J = 15Hz), δ 4-4.3 (m, 1H, H3 '), δ 3.84-3.77 (m, 1H, H4'), δ 3.67 -3.51 (m, 6H, H5 ', 4-pyrrolidine H's) δ 2.24 (t, 2H, H 2', J = 6.08Hz), δ 2.14 (s, 3H, 5- CH ₃ ), δ 1.87-1.78 (m, 4H, pyrrolidine).

Analysis for C ₁₄ H ₂ oN ₆ 0 ₃ :

Calculated: C, 52.49%, H, 6.29%, N, 26.23%; Found: C 52.37%, H 6.33%, N 26.17%.

Example 60 1- (3'-azido-2 ', 3'-dideoxy-β-D-erythro-pentofuranosyl) -4-benzyloxy-5-methyl-2- (1H) -pyrimidinone 5'-acetyl-3'azido-3'deoxy-4- (1,2,4-triazolyl) -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 Butoxy (2 equivalents) in dry acetonitrile added. 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 CHCl ₃ / EtOAc (1: 1, v / v) and the resulting crystals were filtered off to give the title compound. Melting point = 133 ° C to 134 ⁰ C.

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

H ¹ NMR (DMSO-d ₆ ): δ 8.05 (s, 1H, H6), δ 7.45-7.35 (m, 5H, phenyl), δ 6.07 (t, 1H, H1 ', J = 5.94Hz), δ 5.35 (s, 2H, benzyl), δ 5.27 (t, 1H, 5'OH, J = 5.20Hz), δ 4.41-4.31 (m, 1H, H3 '), δ 3.91-3.83 (m, 1H, H 4'), δ 3.7-3.6 (m, 2H, H 5 '), δ 2 , 4-2.3 (m, 2H, H 2 '), δ 1.9 (s, 3H, 5-CH ₃ ).

Calculated: C 57.14%, H 5.36%, N 19.60%; Found: C 57.06%, H 5.37%, N 19.58%.

Example 61 5'-Acetyl-3'-azido-5-bromo-2 ', 3'-dideoxyuridine 3'-azido-2', 3'-dideoxyuridine was prepared according to the procedure of Visser, Synthetic Procedures in Nucleic Acid Chemistry, 1.410 acetylated and brominated to give the title compound. Melting point = 112 ° C to 114 ° C.

UV (nm): at pH 1Amax = 279.210 (ε = 9500.10600),

Amine = 243 (ε = 2000); at pH 13Amax = 276 (ε = 700), amine = 250 (ε = 4000).

H ¹ NMR (DMSO-de): δ 11.88 (s, 1 H, NH) δ 8.02 (s, 1 H, H 6), δ 6.08-6.01 (m, 1 H, H 1 ' δ 4.47-4.40 (m, 1H, H3 '), δ 4.27-4.24 (m, 2H, H 5'), δ 4.03-4.00 (m, 1 H, H4 '), δ 2.41-2.34 (m, * 2H, H 2'), δ 2.09 (s, 3H, A * <rtyl).

Analysis for C ₁₁ Hi ₂ N ₅ O ₅ Br: Calculated: C 35.31%, H 3.23%, N 18.72%, Br 21.36%; Found: C 35.29%, H 3.25%, N 18.66%, Br 21.45%

Example 62 i-O'Azido ^ ', S'-dideoxy-β-D-threo-pentofuranosyl-S-itrifluoromethyl-uracil

The 5'-hydroxyl group of 5-trifluoromethyl-2'-deoxyuridine was protected with a triphenylmethyl group and the 3'-hydroxyl group mesylated. The product (3.0 g, 4,9mMol) was reacted at 70 ⁰ C with 0.7 g lithium azide in 50 ml of DMF over a period of about 26 hours. The cooled reaction mixture was poured onto ice with stirring. 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 0.83 g of the tritylated product. This was dissolved in a saturated solution of zinc bromide in acetonitrile and stirred at 0 ^° C. overnight. After adding 100 ml of ammonium acetate (1 molar), the organic layer was separated and evaporated to dryness in vacuo. The residue was purified by flash chromatography using CHCl ₃ / CH ₃ OH (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 gmol, 5.3%. Melting point = 118 ° C to 120 ° C.

Analysis for C ₁₀ H ₁ OF ₃ N ₅ O, ^

Calculated: C 37.39%, H 3.14%, N 21.80%, F 17.74%;

Found: C 37.31%, H 3.23%, N 21.75%, F 17.60%.

Example 63.

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

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 the 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 ⁰ C.

Example 64 1- (3'-azido-2 ', 3'-dideoxy-β-D-erythro-pentofuranosyl) uracil

The 5'-hydroxyl group of 2'-deoxyuridine (30 g, 0.13 mol) was prepared by the method of Horwitz et al., J. Org. Chem., 31, 205 (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 recrystallized first from ethyl acetate / methanol and then in water. The recrystallized 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 to 168.5 ⁰ C ⁰ C.

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

5-Chloromercuri-2'-deoxyuridine was prepared from 2'-deoxyuridine (10g, 0.044 mol) by the method of Bergstrom and Ruth, J. Carb. Nucl. and Nucl., 4,257 (1977). 2'-Deoxy-5-ethyluridine was obtained 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'-hydroxy group was unmasked by the method of Example 64. The 3'-chloro of threo-3'-chloro-2 ', 3'-dideoxyuridins was with inversion of configuration by heating in HMPA with lithium azide (5 equivalents) displaces at 55 ⁰ 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. Melting point = 112.5 ° C to 115 ° C.

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

5-Bromovinyl-2'-deoxyuridine (BVDU) was synthesized according to the method of Jones et al., Tetrahedron Letters, 45, 4415 (1979) in similar yields. 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 0,955g (19.5 mmol) of lithium azide and heated at 74 ⁰ 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.

Calculated: C 35.98%, H 3.57%, 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-methylisoctosine

1- (3'-azido-2 ', 3'-dideoxy-β-D-threo-pentofuranosyl) -2-methoxy-5-methyl-4 (H) -pyrimidinone (0.5g, 2 mmol) was placed in a bomb with 15 ml of methanol saturated with ammonia. After 6 days at room temperature, the bomb was heated to 65 ^° C. in an oil bath for 4 days. 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 Ci ₀ H ₁₄ N ₆ O _3. ¹ AH ₂ O: Calculated: C 44.36%, H 5.40%, N 31.04%; Found: C 44.42%, H 5.36%, N 31.04%.

Example 68

1 - (3'-azido-2 ', 3'-dideoxy-β-D-threo-pentofuranosyl) -3-methyl-thymine

5'-Trityl-3'-threo-3'-azido-3'-deoxythymidine (1.0 g, 1.95 mmol) and Ν, Ν-dimethylformamide-dichloroacetal (Zemlicka, Coil, Czech Chem. Comm., 35, 3572 [1972]) (0.93 g, 7.8 mmol) were refluxed in 50 ml 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): at pH 1 \ max = 267 (ε = 8100), Ash = 209

(ε = 8000); at pH 13 Amax = 267 (ε = 8000).

H ¹ NMR (DMSO-de): δ 7.56 (s, 1 H, H 6), δ 6.07 (dd, 1 H, H ₁ ,), δ 3.17 (s, 3H, N-CH ₃ ), δ 1.86 (s, 3H, 5-CH ₃ ). Analysis for C ₁₁ H ₁₅ N ₅ O ₄ .0.4 HOAc. 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 (E) -3- [1- (3'-azido-2 ', 3'-dideoxy-β-D-erythro-pentofuranosyl) -1,2,3,4-tetrahydro-2,4-dioxo 5-pyrimidinyl] -2-acrylic acid 2'-Dioxyuridine was converted to the 5-chloromercurri 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 of dry methanol containing ethyl acrylate (104 g, 1.04 mol) and a 0.1 N solution of Li ₂ PdCl ₄ 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 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 solid. This product was tritylated and mesylated by the method of 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 vortexed with water, then filtered and air dried to give 6.1 g of a white solid. 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 at 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 Amax = 300 (ε = 20400),

Xmin = 230 (ε = 3900), = 262 (ε = 13100); at pH 13 Amax = 297.266 (ε = 15 600.14800), amine = 281, 288 (ε = 13600.8600).

H ¹ NMR (DMSO-de): δ 8.37 (s, 1H, H6), δ 7.30 (s, 1H-CH =, 5 = 15.57Hz), δ 6.78 (d, 1 H, = CH-COOH, 5 = 15.93 Hz), δ 6.1-6.06 (m, 1H, HT), δ 5.41-5.37 (m, 1H, 5ΌΗ), δ 4.47-4.39 (m, 1H, H3 '), δ 3.87-3.83 (m, 1H, H4), δ 3.74-3.58 (m, 2H, H 5' ), δ 2.54-2.81 (m, 2H, H2 ').

Analysis for C ₁₂ H ₁₃ N ₅ O ₆ :

Calculated: C 44.59%, H 4.05%, N 21.67%; '

Found: C 44.45%, H 4.06%, N 21.60%.

Example 70 (E) -3- [1- (3'-azido-2 ', 3'-dideoxy-β-D-threo-pentofranosyl) -1,2,3,4-tetrahydro-2,4 dioxo-5-pyrimidinyl] -2-acrylic acid 2'-Deoxyuridine was prepared according to the literature procedure of Bergstrom and Ruth, J. Carb. Nucl., 4,257 (1977) was converted to the 5-chloromercuri derivative in 96% yield. This product (50 g, 1.04 mol) was dissolved in 800 mL of dry methanol containing ethyl acrylate (10.4 g, 1.04 mol) and a 0.1 N solution of Li ₂ PdCl ₄ Jn methanol (10.40 mL) , The solution was stirred for 4 hours and then treated with hydrogen sulfide for 2 minutes. The suspension was passed through a

Celite pad filtered and the filtrate evaporated to dryness in vacuo. The residue was triturated with methanol to give a solid which was filtered off and dried on air to yield 22 g of a white solid. This product was tritylated and mesylated by the method of Horowitz et al., J. Org. Chem., 31, 205 (1966). A portion of this material (2.7g, 4.2mmol) was dissolved in dry DMF (55ml) containing lithium azide (0.62g, 12.7mmol) and heated to 70 ° C under nitrogen for 24 hours. The reaction mixture was poured onto 400 ml of ice, the precipitate collected and purified by flash chromatography. Elution with CHClA / MeOH (50: 1, v / v) afforded 1.48 g of the azido intermediate. This material was treated with 50% acetic acid (100 ml) at 100 ° 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.

UV (nm) at pH 1 Amax = 301 (ε = 19500),

Amine = 230 (ε = 3600),

ASchulter = 249 (ε = 12700); at pH 13 Amax = 299.267 (ε = 1400.13200),

Amine = 232.239 (ε = 1 200.7560).

H ¹ NMR (DMSO-de): δ 3.13 (s, 7H, H6), δ 7.32 (d, 1 H-CH =, δ = 15.87Hz), δ 6.78 (d, 1H , = CH-COOH, J = 15.62Hz), δ 6.01-5.98 (m, 1H, H1 '), δ 5.11-5.98 (m, 1H, 5'-OH) , 4.50-4.43 (m, 1H, H3 '), δ 4.13-4.08 (m, 1H, H4'), δ 3.80-3.75 (m, 2H, H5 '), δ 2.77-3.67 (m, 1H, H 3'), δ 2.30-2.20 (m, 1H, H 2 ').

Analysis for C ₂ H ₁₃ H ₅ O ₆ -1,5H ₂ O: Calculated: C 41.15%, H 4.60%, N 19.99%; Found: C 41.38%, H 4.50%, N 20.01%.

Example 71 (E) -3- [1- (3'-azido-2 ', 3'-dideoxy-β-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 procedure of Example 69. This material was mixed with 10% Pd / C in EtOH to produce the propanoate. This product was then treated in the manner described above to give the title compound, mp = 118 ° to 120 ° C.

UV (nm): at pH 1 \ max = 265 (ε = 9900), AmIn = 235 (ε = 3100); at pH 13 Amax = 265 (ε = 7400), amine = 247 (ε = 5400).

H ¹ NMR (DMSO-d ₆ ): δ 7.68 (s, 1 H, H ₆ ), δ 6.11-6.06 (m, 1 H, HT), δ 4.45-4.35 (m , 1 H, H3 '), δ 3.85-3.80 (m, 1 H, H4'), δ 3.68-3.53 (m, 1H, H5 ').

Analysis for Ci ₂ Hi _S N _s O ₆ :

Calculated: C 44.31%, H 4.65%, N 21.53%;

Found: C44.26%, H 4.68%, N 21.49%.

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 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) t} and lower (C _min ) values in AZT on day 3 (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 (t'l / 2) of AZT (0.88 to 1.73h) during PB treatment. The mean ratio of AZT glucuronide / AZT in urine was significantly reduced from 7.3 to 2.4 after PB treatment. The major parmakokinetic parameters of AZT before and after co-administration of PB are summarized in Table I below.

Table I

patient AUC Cmax Cmin Tm ax Cl, ot / F tl / 2 No. (H * Mm) (Mm) (Mm) (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 0.37 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 ± SD 0.47 0.87 0.00 0.13 480.17 0.01

Example 73 Antiviral activity

The Enhancement of Anti-Friend Leukemia Virus (FLV) Activity of 3'-Azido-3'-Deoxythymidine (AZT) by the Nucleoside Transport Inhibitors Dipyridamole, Dilazep, and 6 - [(4-Nitrobenzyl) -thiol] -9- β-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 the compounds to be tested / combinations giving 50% inhibition of the plaques were determined as shown in Table II. Neither dipyridamole (10μΜ) nor dilazep (5μΜ) alone had any observed antiviral activity.

Table II

Compound / combination azidothymidineED ₅ o (nM)

AZT 5

AZT + 1 μΜ dipyridamole 1

AZT + 5 μΜ dipyridamole 0.5

AZT + 10 μΜ dipyridamole · 0.2

AZT +5 μΜ Dilazep 0.5

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

A patient with a platelet count of 38,000 mm- ³ was diagnosed with a thrombocytopenic purpura (platelet count <100,000 mm- ³ ) and treated with 5 mg / kg AZT intravenously every 6 hours for 6 weeks during which time its platelet count increased to 140,000 mm- ³ .

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 70,000 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 194000 mm ^-3 . A reduction to 2.5 mg / kg / 4 hours orally resulted in a slight decrease in platelet count in an undefined manner but not to extensive diagnostics of ITP.

Example 75 Treatment of Kaposi's sarcoma with 3'-azido-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.

ADV alone showed low activity, a concentration of 16μg / ml (the highest tested) had less than 30% protection, while AZT at 8μΜ showed 100% protection by this method.

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

Table III

ACV ^ g / ml) AZT (μΜ)

- 0

8 0,5

2 1

1 4

0 8

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 RPMI 1640 supplemented with 20% fetal calf serum (FCS), antibiotics, 1-glutamine and 10% interleukin-2 (IL-2) (Electronucleonics, Bethesda, MD). bred. Cells were allowed between 4 to 6 days after exposure to PHA

Concentrations of 4 × 10 ⁵ cells / ml in 25 cm ³ flasks containing 5 ml of medium were distributed and exposed to the action of the agents and the viruses, as detailed below. The day of virus inoculation is referred to 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 and 4 were terminated after 14 days and experiments 1 and 3 after 16 days.

The viral materials were cell-free supernatant fluids from HIV-infected H9 cells, frozen in aliquots at -70 ⁰ C. The 50% tissue culture infectious dose (TCID ₆₀₎ of virus stock was 10 ⁵ / ml.

Four separate experiments were performed using PBMC from different donors (Table IV). in the

Trial 1 indicated both agents 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 to 4, the cells were incubated for 24 hours with or without the medium

Recombinant a-interferon (HFNaA) incubated, then AZT and virus exposed. The viral innocula were 4 × 10 ³ , 10 ³ and 2 × 10 ³ TCID ₅₀ in experiments 2, ³ and 4, respectively. The concentrations of the agents were the same each time the medium changed

adjusted that original concentrations were maintained.

Twice dilutions of a specified combination of rlFNaA and AZT were serially studied in all experiments.

Each concentration of rlFNaA and AZT used in combination was also studied alone for reference

create.

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

Combination, examined.

In experiments 2 and 3, 0.16μΜ AZT and 128U / ml of rlFNaA and 3-4x double dilutions of this combination were used. In experiment 4, 0.08 μΜ AZT, along with 128 U / ml of rlFNaA and 2 double dilutions, were obtained

this combination, 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, virus 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 dielobologram 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 on.

Table IV

attempt

HIV

inoculation

method*

HIV supplement (TCID ₅₀ )

Time of the addition of funds rlFNaA AZT

A B B B

10 ⁵

4x10 ³ 10 ³ 2X10 ³

-24 h -24 h -24 h

* Method A: 40 χ 10 ⁶ cells are suspended in 20 ml of the medium containing 10 ⁶ TClD ₅₀ HIV for 1 hour at 37 ⁰ C, 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 Experiment 2 - Day 10

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

AZT (μΜ)

rIFNaA (U / ml) 16 32

64

128

0 205 173 0.01 1:59 85 0.02 110 0.04 71 0.08 31 0.16 7

150

42

193

10

169

151

Table VI Trial 3-Day 13

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

AZT (μΜ)

16

rlFNaA (U / ml) 32

64

128

157 143 117 51 10 10 0 2 5

117

130

Table VII Experiment 4-day 11 Effects of HFNaA and AZT on mean RT values (cpm / 10 ⁶ cells χ 10 ³ )

rlFNaA (U / ml)

AZT (μΜ)

32

64

128

32 8 4 1

4 0

TABLE VIII EXPERIMENT 3 - DAY 13 Effects of HFNaA and AZT on virus yield (TCIDso / ml)

AZT (μΜ)

16

HFNaA (U / ml) 32

64

128

10 " 10 ^s · ⁶ ₁₀ 4.8 10 ¹ · ⁹ 10 ³ 10 ¹ · ⁹

5.3

_10s , 1

<10 ¹ '· ⁹

Table IX Experiment 4 - Day 14 Effects of rlFNaA and AZT on HIV p24 *

AZT (μΜ)

IFNaA (U / ml)

32

64

128

200-300 100-200 50-100 100-200 2.2 25-30 1.0 11.2

25-30

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

attempt

Day in

Culture

50%

90%

95%

7 10

6 13 16

8 14

2.14 0.34 0.37 0.30 0.26 0.73 0.12 0.15 <0.01 0.02 0.01 0.03 0.02 0.07

0.23 0.28 1.39 0.17 0.05 0.04 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 testing, MIC determinations for each compound were made individually against the test organism (E. coli CN314) Table XI shows the MIC endpoints of each agent for E. coli CN314.

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 value (Table XI).

The test plates were inoculated with a bacterial inoculum containing approximately 5 x 10 ^s 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 of fractions (the sum of the fractional inhibitory concentrations) was then calculated, a result of about 0 , 5 or below indicates synergy.

Table Xl Minimal inhibitory concentrations (MIC values) of the tested agent alone Compound MIC / g / ml)

Tobramycin 0.4

Fusidic acid 1000

Chloramphenicol 3.1

Clindamycin 100

Erythromycin 25

Rifampicin 6.2

3'-azido-3'-deoxythymidine 1.0

Trimethoprim 0.125

Sulfadimidine 32

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

Table XII Synergy studies: combinations of AZT and other antibacterial agents combination

Tobramycin / AZT Fusidic Acid / AZT Chloramphenicol / AZT Clindamycin / AZT Erythromycin / AZT Rifampicin / AZT Trimethoprim / AZT Sulfadimidine / AZT

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

The in vitro activity of 3'-azidonucleosides (drugs) was determined in two cell lines; H9 (OKT4 ⁺ T cell line, which allows HIV replication but is partially resistant to the cytopathic effect of HIV) and TM3 (T cell clone, specific to tetanus toxoid, immortalized by lethally irradiated HTLV) I and selected for rapid growth and sensitivity to the cytopathic effect of HIV).

The inhibition assay was performed as follows: TM3 cells were stimulated by antigen plus irradiated (4000 rads, 40 Gy) fresh autologous peripheral blood mononuclear cells (PBM) and in complete medium containing 15% (v / v) interleukin -2 (IL-2, lectin-depleted; Cellular Products, Buffalo, NY) cultured 6 days prior to testing. ATH8 cells were used without the antigen stimulation. After pre-exposure to 2 μg polybrene per ml for 30 minutes, the target T cells (2 × 10 ^s ) were pellitized, exposed to HIV for 45 minutes, resuspended in 2 ml fresh medium and placed in culture tubes at 37 ^° C. incubated in 5% CO 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 x 10 ⁴ lethally irradiated (10,000 rad) HIVRF-II-producing H9 cells or uninfected H9 cells were added to 2 x 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.

Optimum MIC values (Mg / ml): FIC index Medium / AZT 0.25 0.2 / 0.125 0.28 250 / 0.03 0.31 1.6 / 0.06 0,375 12.5 / 0.25 0.5 6.2 / 0.25 0.56 3.1 / 0.06 0.504 0.004 / 0.5 0,375 0.25 / 0.125

Table XIII

connection

ED ₅₀ (MM)

3'-azido-2 ', 3'-dideoxycytidine 3'-azido-5-bromo-2', 3'-dideoxyuridine 3'-azido-5-bromo-2 ', 3'-dideoxycytidine (E) -3- [ 1- (3'-azido-2 ', 3'-dideoxy-β-D-erythropentofuranosyl) -1,2,3,4-tetrahydro-2,4-dioxo-5-pyrimidinyl] -2-acrylic acid

10 5 5

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 1 2 3 MIC (Mg / ml) 6 7 δ default 0.1 > > 100 10 connection 0.1> 10 10 organism 0.1 §> 10 10 4 5 10 10 > 100 > 100 0.1 » 0.1 => 10 0.1 > 10 10 10 0.1 > 0.5 E. coli 0.1 => 10 > 100 0.1>> 100 10 10 > 100 0.1 Salmonella typhimurium 10 10 10 0.1 > 0.1 > 10 10 > 100 0.5 Salmonella typhosa > 100 100 0.5 Entero. aerogenes 0.1 > 10 Citro.freundii

The compounds used were:

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

2 = S'-azido-S'-deoxy-S'-O-acetyl-A-thiothymidine

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

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

5 = i-IB'-O-acetyl-S'-azido ^ ', S'-deoxy-β-D-erythro-pentofuranosyD-B-methyl-A-d ^^ -triazol-i-yD ^ dHt-pyrimidinone

6 = 1- (3'-A2ido-2 ', 3'-dideoxy-β-D-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 (4)

1. A process for the preparation of a pharmaceutical formulation, characterized in that a) preparing a compound of formula (I) wherein B is a purine or pyrimidine base attached at the 9 or 1 position, or a pharmaceutically acceptable derivative thereof, by a process comprising: (A) a compound of formula III (wherein A is a purine or pyrimidine base attached at the 9- or 1-position, respectively, and M is a precursor moiety for the 3'-azido group) or a derivative thereof, with an agent or under conditions which to convert the precursor moieties to the desired azido group; or (B) a compound of formula (IVa) or (IVb) wherein Rl, Ri Rl, Rt Rf and Rg are 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 linked to form a heterocycle; R ^{2 is} hydrogen, acyl, alkyl, aroyl or sulfonate; R ^{3 is} hydroxy, mercapto, amino, triazolyl, alkylamino, dialkylamino wherein the alkyl groups are optionally joined to form a heterocycle, aralkoxy, alkoxy or alkylthio; R ^{4 is} alkyl, substituted alkyl, halo, 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 ¹ , R ² , R ³ , R ⁴ in the formula (IVa) or at least one of R ⁶ and R ⁷ in the formula (IVb) represents a precursor group with a Means or under conditions which lead to the conversion of the precursor group (s) into the corresponding desired groups; or - (C) a compound of formula (Va) or (Vb) (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, which reacts to introduce the desired ribofuranosyl ring at the 1-position of the compound of formula (IVa) or the 9-position of formula (IVb); or (D) if B of the compound of formula (I) A is a purine residue, reacting a purine of formula (Vb) with a pyrimidine nucleot of formula (VI) (wherein Py represents a 1-pyrimidinyl group); and subsequently or simultaneously with one or more of the following transformations, if appropriate: (i) if a compound of formula (I) A is formed, the compound is converted into a pharmaceutically acceptable derivative thereof, and (ii) if a pharmaceutically acceptable derivative of a compound of formula (I) A is formed, the derivative into the parent compound of the formula (I) A or into another pharmaceutically acceptable derivative of a compound of the formula (I) A; and b) the resulting compound of formula I (A) or a pharmaceutically acceptable derivative thereof interacts with at least one further pharmaceutically active ingredient and a pharmaceutically acceptable carrier. A process according to claim 1, characterized in that a compound of formula (I) A is prepared according to process A wherein B is a thymidine base and the 3'-azido group is in the erythroid configuration. 3. The method according to claim 1 or 2, characterized in that the compound of formula (I) A or a pharmaceutically acceptable derivative thereof with acyclovir (9- (2-HydroxyethoxymethyDguanin) is brought into interaction. 4. The method according to claim 1 or 2, characterized in that the compound of formula (I) A or a pharmaceutically acceptable derivative thereof is brought into interaction with an interferon. ~ For this 3 pages drawings