EP0437469A1

EP0437469A1 - Nucleoside derivatives

Info

Publication number: EP0437469A1
Application number: EP89911035A
Authority: EP
Inventors: Jo Klaveness; Kjell Undheim; Frode Rise; Jostein Hatlelid
Original assignee: Nycomed AS
Current assignee: GE Healthcare AS
Priority date: 1988-10-05
Filing date: 1989-10-05
Publication date: 1991-07-24
Also published as: NO176052C; ES2058481T3; JPH04500968A; DK58791D0; AU617543B2; NO911322L; ZA897594B; FI92491B; NO911322D0; DK58791A; NZ230919A; OA09642A; FI911621A0; DE68918036D1; FI92491C; WO1990004602A1; GB8823320D0; ATE111104T1; DE68918036T2; EP0362967A1

Abstract

Compounds of formula (I) <CHEM> wherein Y<1> is a hydrogen atom or an acyl or acyloxyalkyl group; Z is a hydrogen atom or an azido group; and X is a purine or pyrimidine base or an ester, amide or acyloxyalkyl, derivative thereof. The compounds have antiviral activity.

Description

NUCLEOSIDE DERIVATIVES

This invention relates to antiviral compounds and more particularly to esters and amides of nucleoside derivatives which are active against human immuno¬ deficiency virus (HIV) , the retrovirus which causes the disease AIDS.

AIDS is a relatively new disease. It was discovered in 1981 and several thousand cases of the disease have been diagnosed since then. It is anticipated that the number will increase to at least several hundred thousand in the next few years. The situation is especially severe in several Central African countries. AIDS is fatal, and about 40% of all diagnosed cases have ended in death. Of those diagnosed as having AIDS three or more years ago it is estimated that 85% are now dead.

Clinical symptoms are weight loss, chronic diarrhoea, persisting fever and opportunistic infections due to loss of T-cells, thus upsetting the overall balance of the immune system. The patient loses his/her ability to combat otherwise insignificant infections.

Several different methods to combat the infection have been tried. Among the methods tried are stimulation of the immune system and conventional treatment of the (secondary) life-threatening infections. So far the most promising method has been to attack the replication of the HIV-virus. Several different compounds interfering with replication have been tried, e.g. phosphonoformate (Foscarnet) , suramin, Evans Blue, 3'-azido-3 '-deoxythymidine (AZT) and 2¹, 3 '-dideoxynucleosides.

European Patent Application No. 0196185A, for instance, describes pharmaceutical compositions containing AZT, a known compound which has shown great promise in the treatment of AIDS and AIDS- related complex. It is believed that AZT works by inhibiting reverse transcriptase, a vital enzyme in the life cycle of retroviruses.

Further work has been done on alternative reverse transcriptase inhibitors which might avoid the limitations and drawbacks of AZT, for instance bone marrow suppression or the need for frequent administration of relatively large quantities, and among those suggested have been the 2' ,3'-dideoxy- nucleosides.

The synthesis and activity of these compounds have been described (Mitsuya and Broder, Proc. Natl. Acad. Sci. 3, 1911 (1986)) and it was demonstrated that both the 2' and 3' positions must be unsubstituted while the 5'-hydroxy group must be present, presumably to allow j vivo conversion to the corresponding nucleotides.

European Patent Application No 0206497A discloses 2' ,3'-dideoxyribofuranoside derivatives of cytosine or purine bases as antiviral compounds. While there is reference to esters of these compounds as possible metabolic precursors, there is no suggestion that esters would possess any advantageous properties compared with the parent 5'-hydroxy compounds and no esters are specifically named or their synthesis exemplified. There is no reference to any corresponding thymidine compounds or of any nucleoside derivatives having N-acylated amino groups.

We have now found that esterification or etherification of the 4- or the 5'- oxygen and/or amidation of exocylic or endocyclic nitrogen atoms present in the purine or pyrimidine ring can give significant advantages in terms of uptake, overall activity and site of action. Our copending PCT Application PCT/GB88/00224 describes certain esters and amides of this type carrying acyl groups at the 5' position or on exocyclic nitrogens; the - 3 - present invention extends this principle to a wider range of related compounds.

Thus according to one feature of the invention we provide compounds of formula (I)

(wherein Z is a hydrogen atom or an azido group,

Y is a hydrogen atom or a physiologically acceptable group of the formula R¹(O)_nCO(OCR²R³) -

where n is 0 or 1, m is 0 or 1 and

R is an optionally substituted alkyl or aryl group or, where n is 0, a hydrogen atom. R 2 and R3 are independently hydrogen atoms or lower alkyl groups; and

X is a group selected from

(F) - 4 -

(where the groups Y"¹ and Y are as defined for Y and may be the same as or different from

Y 1 or each other, R4 is a hydrogen atom or a group -NY 3Y4, where Y3 and Y4 have the above meanings and R is a hydrogen atom or a lower alkyl group, with the proviso that at least one of the groups

1 2 3 4

Y , Y , Y and Y is other than hydrogen, and that

2 when Y is absent or is a hydrogen atom and any groups Y 1, Y3, and Y4 which are present are either hydrogen atoms or groups R 1 (O) CO(OCR2R3) where m is 0 then Z is an azido group) and/or salts thereof.

It will be appreciated that some of the groups

2 X, for example those in which Y is a hydrogen atom, are tautomers of other of the groups X and exist in equilibrium with them

According to a further feature of the invention we provide for the use of compounds of formula (I) as hereinbefore defined, and/or salts thereof, in the manufacture of a composition for the treatment or prophylaxis of retrovirus infections, in particular neurotropic viruses and especially HIV infections. Such compositions also form part of the invention.

The group R is preferably an alkyl group containing 1-20 carbon atoms which may be straight or branched, or an aryl group which may contain 6 to 20 carbon atoms and may be mono- or poly-cyclic. Substituents which may be present on the alkyl groups R include aryl groups preferably having 6-10 carbon atoms (as in aralkyl groupings) , hydroxy and carboxy groups. Aryl groups include heterocyclic aryl groups such as pyridinyl and thienyl groups.

Substituents which may be present on aryl groups include alkyl groups, e.g. having 1-6 carbon atoms, hydroxy and carboxy groups. Examples of such groups include methyl, ethyl, propyl, t-butyl, pentyl, stearyl, palmityl, carboxyethyl and benzyl groups.

The lower alkyl groups R 2, R3 and R5 preferably

2 contain 1-6 carbon atoms. However, R preferably represents a hydrogen atom. R 3 i.s preferably a hydrogen atom or more preferably a methyl group

5 R is preferably a hydrogen atom or a methyl group.

It will be noted that the compounds of the invention may carry more than one of the groups Y 1, Y2, Y3 and Y4. In the compounds of formula

(I) D,E, I and J, it is preferred that m in the group Y 4 is 0 (zero) . Groups Y2 are preferably of the formula

R¹.CO-, R¹CO.O.CR²R³ or R^-.O.CO.O.CR^³-.

The salts of the compounds of formula (I) may be acid addition salts with organic or inorganic acids, for instance hydrochloric or phosphoric acid or methanesulphonic acid, ethane disulphonic acid, 2-naphthylsulphonic acid, pivalic acid and pamoic acid. Antiviral counter-ions such as phosphono- formate or suramin may also be used. Organic or inorganic base salts may be formed with acidic groups present in the molecule; suitable counter- ions include alkali metal ions such as sodium and potassium ions, divalent ions such as calcium and zinc ions and organic ions such as tetraalkylammonium and choline or ions derived from meglumine or ethylene- diamine. Salts according to the invention may be formed by reaction of the compound of formula (I) with an appropriate acid or base.

The compositions according to the invention may be used in the treatment and/or prophylaxis of retrovirus infections, in particular HIV infections, and such a method forms a further feature of the - 6 - invention. They may be formulated in conventional manner by admixture of one or more compounds of formula (I) as defined above with excipients and/or carriers.

It is believed that the compounds of formula (I) are not themselves inhibitors of reverse transcriptase but are converted J i vivo to the 5-hydroxy-2' ,3'- dideoxynucleosides. Nevertheless the substitution at the respective 0- and N- atoms gives surprising advantages in terms of uptake and sustained activity. The compounds of formula (I) are more lipophilic than the parent compounds and this permits rapid and efficient absorption from the gastro-intestinal tract; the absorption rate may be optimised by careful choice of the substituent group to give the desired balance of lipophilicity and hydrophilicity. The lipophilic nature of the compunds of formula (I) also gives the molecules the ability to penetrate the cell membranes more easily and leads to higher intracellular concentrations, giving an improved dose/effect ratio. The steady hydrolysis of the compounds ensures a sustained concentration of the active compound in the cell and thereby permits longer intervals between doses, overcoming a significant drawback of the prior art compounds such as AZT.

Finally, the compounds according to the invention can penetrate the blood-brain barrier and thus permit treatment of the neurological disorders which have been observed to be related to the presence of neurotropic viruses, e.g. retroviruses such as HIV, and lentiviruses (Yarchoan et al, The Lancet, January 17, 1987, page 132). This is a significant advantage compared to the corresponding unsubstituted compounds or other antiviral compounds and is not referred to anywhere in the prior art, for instance in EP-A-0196185 or in EP-A-0206497. Attempts have been made to treat these neurological disorders with AZT but with limited success. The invention thus further provides a method of treatment of neurological disorders caused by neurotropic viruses wherein an effective dose of a compound of formula (I) or a salt thereof is administered to a patient suffering from such a disorder.

Compounds of formula (I) may be prepared in any convenient way, for example, by reaction of a compound of formula (II)

[wherein Y and Z are as hereinbefore defined and

X is as hereinbefore defined for X except that any of the groups Y 1, Y2, Y3 and Y4 may each additionaly represent a protecting group, with the proviso that at least one of Y 1, Y2, Y3 and Y4 is a hydrogen atom] with a reagent serving to introduce a group

R 1(0)'nCO. (OCR2R3)m as defined above followed where required by removal of any protecting groups and/or unwanted substituents so introduced.

It should be noted that where, in the starting material, more than one of Y 1, Y2, Y3 and Y4 is hydrogen, multiple reactions may occur.

Where it is desired to ensure that acylation or alkylation is effected while one or more groups

Y , Y , Y and Y remain as hydrogen atoms, it may be desirable to protect the latter first, to form a compound of formula (I) in which one or more of Y 1, Y2, Y3 and Y4 are protecting groups, these being removed after introduction of the desired acyl or ether group. Such protecting groups may, in fact, be conventional N- or O-protecting groups including groups R OCO- which may be selectively removed in the presence of the group(s) intended to remain. Thus, for example, an N-benzyloxycarbonyl may be used to protect an exocylic amino group and if the group which is intended to remain is not one which is removable by reduction, for example a straight chain alkoxycarbonyl group, the N-benzyloxycarb group can readily be removed selectively using hydrogen and a noble metal catalyst such as palladium.

Trisubstituted silyl groups may also be used as protecting groups, especially for the 5'-oxygen atom, and include trialkylsilyl e.g. trimethylsilyl, dimeth 1-t-butylsilyl, and thexyldimethyl silyl groups. In general, where more than one of Y 1, Y2,

Y and Y are hydrogen, and a mixture of compounds is produced, the individual components may readily be separated, for example by chromatography.

Where 5'-0-monoalkylation is to be effected (i.e. introduction of a group Y in which m is 1) it is especially effective to form a dianion of the nucleoside (e.g. by reacting with sodium hydride) and to react this with one equivalent of the alk lating agent. It is of course, still possible to use protected forms of the nucleoside, for example by acylation of a nucleophilic nitrogen atom before salt formation with sodium hydride.

Suitable acylating agents for use in the reaction have the formula Ac-L where L is a leaving group. When the acyl group Ac- is derived from a carboxylic acid, i.e. is of formula R -CO-, then suitable acylating agents include the acid halides and acid anhydrides advantageously in the presence of a base; when the acyl group is derived from a carbonic acid, i.e. is of formula R .0.C0-, then acylating agents include the haloformate esters and reactive carbonic acid diesters. In such reagents, the halogen may, for example, be chlorine or bromine. The base for use in the reaction with the acid halide or anhydride may, for example, be a heterocyclic base such as pyridine or 4-dimethylaminopyridine. The latter increases the speed of the reaction and may be used advantageously with pyridine. The reaction will normally be carried out in the presence of an inert solvent e.g. a substituted amide solvent such as dimethylformamide, dimethyl- acetamide or a halogenated hydrocarbon such as dichloromethane.

In general, we have found that using acid anhydrides as acylating agents to introduce a group R CO, O-acylation at the 5¹- position takes place more readily than N-acylation, whereas using acid halides, N-acylation or even N-diacylation predominates. However, N-acyl groups R CO- may be removed selectively, for example by reaction with a phenol such as p- methyl-phenol. where multiple substitution is to be effected, a stronger base such as sodium hydride may be advantageous.

Suitable acyloxyalkylating agents for use in the invention will in general be of the formula R¹C0.0.CR²R³L or R¹0.C0.0.CR²R³L, where L is a leaving group. Thus, the group L may for example, be a halogen atom such as a chlorine or bromine atom or a hydrocarbon-sulphonyloxy group such as a tosyloxy or mesyloxy group.

The alkylation reaction will normally be effected in the presence of a base, conveniently an inorganic carbonate such as potassium carbonate or an alkali metal hydride such as sodium hydride. Bases as used for acylation may also be useful.

The starting compounds of formula (II) wherein

Y 1, Y2, Y3 and Y4 are all hydrogen atoms are well described in the literature - see, for instance,

Lin et al, J. Med. Chem. :3), 440 (1987). Starting compounds wherein one or more of Y 1, Y2, Y3 and ₄

Y are other than hydrogen may be prepared by preliminary reactions as described above. The pharmaceutical compositions according to the invention may be formulated conventionally by means well known in the art, and may be administered by any convenient route, for instance orally, rectally, vaginally, intraveneously or intramuscularly. Examples of suitable formulations include tablets and capsules, aqueous formulations for intravenous injection and oil-based formulations for intramuscular injection. Suitable dosages will lie in the range 0.1 to lOOmg per kilogram of bodyweight per 24 hour period. The compositions according to the invention may also contain other active antivirals for instance acyclovir, phosphonoformate, suramin, Evans Blue, inte ferons or AZT.

The invention is illustrated by the following Examples (starting materials are either known materials or are prepared according to our copending PCT Application PCT/GB88/00224) . Capsugel is a Trade Mark.

Example 1

3-Acetoxymethyl-2' ,3 '-dideoxyuridine (Formula (I)A, Y² = acetoxymethyl, R⁵=Y¹=H)

A mixture of 2' ,3'-dideoxyuridine (1 mmol) and potassium carbonate (0.8 mmol) in dry N,N-dimethylacet- amide (20 ml) is stirred at ambient temperature for 2 hours, a solution of acetoxymethyl chloride (0.7 mmol) in dry N,N-dimethylacetamide (5 ml) added dropwise with stirring, the mixture stirred at ambient temperature for 36 hours, filtered, the filtrate evaporated and the product purified by flash chromatography on silica gel using ethyl acetate:light petroleum.

Example 2

3,5'-O-Bis (acetoxymethyl)-2' ,3'-dideoxyuridine

( :FFormula (I)A, Y¹=Y² = acetoxymethyl, R⁵=H) and A 5 '-O-bis (acetoxymethyl-2' ,3 '-dideoxyuridine (Formula (I)B,Y¹=Y² = acetoxymethyl, R⁵=H)

A mixture of 2' ,3'-dideoxyuridine (1 mmol) and sodium hydride (2 mmol) in dry DMF (50 ml) is stirred at ambient temperature for 2 hours, a solution of acetoxymethyl chloride (2 mmol) in dry DMF (10 ml) added dropwise with stirring, the mixture stirred at ambient temperature for 24 hours, filtered, the filtrate evaporated and the product mixture separated by flash chromatography on silica gel using ethyl acetate:light petroleum. Example 3

3-Acetoxymethyl-5 O-palmitoyl-2 * , 3 ' -dideoxythymidine

(Formula ( I) A, Y 2 = acetoxymethyl , Y 1 =palmitoyl,

R⁵=methyl)

A mixture of 5'-O-palmitoyl-2' ,3'-dideoxythymidine (1 mmol) and potassium carbonate (0.8 mmol) in dry N,N-dimethylacetamide (20 ml) is stirred at ambient temperature for 2 hours, a solution of acetoxymethyl chloride (0.7 mmol) in dry N,N-dimethy- lacetamide (5 ml) added dropwise with stirring, the mixture stirred at ambient temperature for 24 hours, filtered, the filtrate evaporated and the product purified by flash chromatography on silica gel using ethyl acetate:light petroleum.

Example 4

N 4,5'-O-Di(benzyloxycarbonyl)-N4-pivaloyloxymeth l-

2' ,3'-dideoxycytidine (Formula (I)C, Y =γ benzyloxycarbon

3 Y = pivaloyloxymethyl) and 3-pivaloyloxymethyl- N 4,5'-0-di(benzyloxycarbonyl) 4-imino-3,4-dihydro-

I I

2 ,3 - dideoxycytidine

(Formula (I) D, Y4 = benzyloxycarbonyl)

4 N ,5'-0-Dι(benzyloxycarbonyl)-2' ,3'-dideoxycytidine

(1 mmol) is dissolved in dry DMF (20 ml) , the solution cooled to 0°C, sodium hydride (1.1 mmol) added, the mixture stirred at 0°C for 1 hour when the bubbling of hydrogen has ceased, pivaloyloxymethyl chloride (1.1 mmol) in dry DMF (10 ml) added dropwise, the resultant mixture stirred at ambient temperature for 36 hours, most of the solvent removed below

40°C using an oil pump, water added to the residue and the mixture extracted with ethyl acetate. The dried (MgSO.) ethyl acetate solution is evaporated and the residue subjected to flash chromatography using light petroleum: ethyl acetate which elutes the title compound.

Example 5

dine (Formula

N 4,5'-O-Di(benzyloxycarbonyl)-N4-pivaloyloxymethyl-

2' ,3 '-dideoxycytidine (0.2 mmol) is added to a suspension of 5% palladium on charcoal (25 mg) in ethanol (25 ml) , the air removed under vacuum and the mixture hydrogenated at atmospheric pressure until the hydrogenolysis is completed (ca. 3 hours) .

The mixture is then filtered, the filtrate evaporated at reduced pressure and the product purified by filtration (chromatography) on neutral silica gel using chloroform:ethanol.

Example 6

3-Pivaloyloxymethyl-3'-deoxythymidine

i) 3'-Deoxythymidine (0.0449 g, 0.198 mmol) and imidazole (0.0328 g, 0.482 mmol) were dissolved in DMF (0.5 ml). Thexyldimethylsilyl chloride (0.047 ml, 0.238 mmol) was added and the reaction mixture was stirred at room temperature for 19 hours. The solvent was removed under reduced pressure and chloroform (15 ml) was added to the residue, washed with water (5ml x 2) , dried (MgSO.) and evaporated. The residue was chromatographed on silica using ethyl acetate-hexane (7:3) ; yield 0.0578 g (79%) of 5'-Q-thexyldimethyl-silyl-3'- deoxythymidine as a white powder. Mp. 109-110°C (uncorrected) .

^^■H NMR(CDC1₃, 300 MHz) *b :0.14(s, 6H) ,0.86-0.90 (m, 12H) ,1.64 (m,lH) ,1.91(s,3H) , 1.93-2.02 (m,3H)2.26- 2.41(m,lH) ,3.66-3.96(ABX,2H) ,4.08-4.17(m,lH) ,6.05(dd,lH) , (d,lH) ,8.50(b,lH) .

13 C NMR(CDC1₃) :-3.51, -3.43, 12.47, 18.31, 18.41,

20.12, 20.32, 25.32, 25.38, 32.30, 33.91, 64.24, 80.76, 85.68, 110.18, 135.53, 150.14, 163.59.

ii) The 5'-0-thexyldimethylsily1-3'-deoxythymidine (0.0254 g, 0.0689 mmol) and potassium carbonate (0.0111 g, 0.0803 mmol) were suspended in DMF (1ml) and stirred for 1.5 hours at room temperature. The mixture was cooled to 0°C and chloromethyl pivalate (0.013 ml, 0.0895 mmol) was added. The mixture was stirred at room temperature for 18 hours before the solvent was evaporated under reduced pressure. The residue was chromatographed on silica using ethyl acetate - hexane (35:65) ; yield 0.031 g (93%) of 3-pivaloyloxymethyl-5'-0-thexyldimethylsilyl- 3'-deoxythymidine as a glassy material. ¹H NMR(CDC1₃, 300 MHz) . 0.14 (s,6H) ,0.87-0.90 (m,12H) ,

1.17 (s,9H) ,1.56-1.70 (m,lH) ,1.94 (d,3H) ,1.95-2.02 (m,3H) ,2.30-

2.44 (m,lH) ,3.67-3.97 (ABX,2H) ,4.11-4.17 (m,lH) ,5.90-

5.98 (AB,2H) ,6.06(dd,lH) ,7.54(d,lH) .

¹³C NMR(CDC1₃) S : -3.49, -3.42, 13.09, 18.31, 20.12,

20.33, 25.34, 26.92, 32.37,33.91, 38.71, 64.22,

64.90, 80.92, 86.35, 109.37, 134.50, 150.23, 162.58,

177.43.

iii) The 3-pivaloyloxylmethyl-5 '-O-thexyldimethylsilyl- 3 '-deoxythymidine (0.029 g, 0.061 mmol) was dissolved in THF (0.5 ml) and a 0.25 M solution of tetrabutylammoniu fluoride in THF (0.5 ml) was added. The mixture was stirred for 30 minutes before the solvent was evaporated. The residue was dissolved in chloroform (7 ml) and washed with water (1 ml) . The organic phase was dried (MgSO.) , evaporated and the residue purified by preparative TLC. The plates were eluted with diethyl ether (3x) and the product extracted from the main band by chloroform-methanol (9:1) ; yield 0.0170 g (83%) of the title compound as a white solid. Mp. 64-66°C (uncorrected)

^~~Tl NMR(CDC1₃, 300MHz)c> : 1.17 (s,9H) ,1.93 (d,3H) ,1.95- 2.15 (m,4H) ,2.32-2.48 (m,lH) ,3.72-4.03 (ABX,2H) ,4.14- 4.21 (m,lH) ,5.90-5.97 (AB,2H) ,6.10(dd,lH) ,7.60 (d,lH) . ¹³C NMR(CDC1₃) £^": 13.12, 24.91, 26.90, 32.22, 38.71, 63.42, 64.89, 81.10, 86.73, 109.62, 135.04, 150.26, 162.51, 177.47.

Example 7

3,5 '-0-Di (pivaloyloxymethyl)-3 '-deoxythymidine

3 '-Deoxythymidine (0.050 g, 0.221 mmol) was dissolved in DMF (1 ml) and cooled to 0°C. Sodium hydride (0.0147 g, 80% in oil, 0.49 mmol) was added and the mixture stirred at 0°C for 1 hour before chloromethyl pivalate was added. The mixture was stirred at 0°C for 1 hour and then at room temperature for 20 hours. TLC indicated partial conversion to the dialkylated product and the mixture was cooled to 0°C and sodium hydride (0.0027 g, 80% in oil, 0.090 mmol) was added. The ixure was stirred at 0°C for 1 hour before chloromethyl pivalate

(0.013 ml, 0.090 mmol) was added. The mixture was stirred for 1 hour at 0°C and at room temperature for 19 hours before the solvent was evaporated under reduced pressure. Chloroform (10 ml) was added to the residue and the mixture washed with saturated sodium chloride (5 ml x 2) and water

(5 ml x 2) . The chloroform phase was dried (MgSO,) , evaporated and the residue subjected to preparative TLC. The plates were eluted with diethyl ether - pentane - methanol (50:50:0.1) (7x) and the product extracted from the main band by chloroform - methanol; yield 0.042 g (42%) as a colourless oil.

H NMR (CDC1₃, 300 MHz) : 1.19 (s, 9H) ,1.24 (s,9H) ,1.95- 2.10 (m,6H) ,2.30-2.50 (m,lH) ,3.74-4.06(ABX,2H) ,4.20- 4.30 (m,lH) ,5.33-5.40 (AB,2H) ,5.92-5.99(AB,2H) ,6.10- 6.16(M,1H) ,7.66(d,lH) .

¹³C NMR (CDC1₃) £ : 13.04, 25.13, 26.91, 32.51, 38.70, 38.84, 64.87, 70.16, 79.42, 86.40, 88.49, 109.44, 134.83, 150.24, 162.53, 177.39, 177.75.

Example 8

5 '-O-Pivaloyloxymethyl-3'-deoxythymidine

A mixture of 3'-deoxythymidine (0.0103 g, 0.046 mmol) and sodium hydride (0.0029 g, 80% in oil, 0.097 mmol) in DMF (0.5ml) was stirred at 0°C for 1 hour. Chloromethyl pivalate (0.0070 ml, 0.048 mmol) was added and the mixture stirred at 0°C for 3 hours. Ammonium chloride 1M (3 ml) was added and the mixture extracted with diethyl ether (3 ml) . The ether extract was washed with saturated sodium chloride (3 ml x 2) , dried (MgSO.) and evaporated. The residue was chromotographed on silica using chloroform - methanol; yield 0.0068 g (43%) as a glassy material.

¹H NMR (CDC1₃, 300 MHz) £: 1.23 (s,9H) ,1.90-2.18(m,6H) , 2.30-2.50 (m,lH) ,3.73-4.05(ABX,2H) ,4.18-4.28 (m,lH) ,5.33- 5.39(AB,2H) ,6.08-6.14(m,lH) ,7.62(d,lH) ,8.44 (b,lH) . ¹³C NMR (CDC1₃) 9: 12.59, 25.29, 27.03, 32.58, 38.97, 70.33, 79.40, 85.83, 88.60, 110.37, 136.01, 150.26, 163.69, 177.92.

Example 9

5'-O-Pivaloyloxymethyl-3'-azido-3'-deoxythymidine

A mixture of 3'-azido-3'-deoxythymidine (0.0335 g, 0.125 mmol) and sodium hydride (0.0079 g, 80% in oil, 0.263 mmol) in DMF (1.5 ml) was stirred at 0°C for 1.5 hours. Chloromethyl pivalate (0.020 ml, 0.138 mmol) was added and the mixture stirred at 0°C for 1 hour. Acetic acid (0.0072 ml, 0.126 mmol) was added and the solvent evaporated. The residue was chromatographed on silica using chloroform

- methanol; yield 0.031 g (63%) as an oil.

¹H NMR(CDC1₃, 300 MHz) S : 1.22(s,9H) ,1.94 (d,3H) ,2.20- 2.50 (m,2H) ,3.76-4.02(m,3H)4.22-4.32(m,lH) ,5.34 (s,2H) , 6.23 (t,lH) ,7.44 (d,lH) ,9.35 (b,lH) .

¹³C NMR(CDC1₃) : 12.51, 26.91, 37.71, 38.90, 59.91, 68.49, 82.58, 84.55, 88.27, 111.32, 135.39, 150.38, 163.88, 177.88.

Example 10

4 —Benzyloxycarbonyl-5 '-0-pιvaloyloxymethyl-2' ,3 '- dideoxycytosine

4 A mixture of N -Benzyloxycarbonyl-2' ,3'-dideoxycytosine

(0.022 g, 0.0637 mmol) and sodium hydride (0.0044 g 80% in oil, 0.147 mmol) in DMF (1.5 ml) was stirred at 0°C for 1.5 hours. The mixture was cooled to -50°C and chloromethyl pivalate (0.0102 ml, 0.0702 mmol) was added. The mixture was stirred at -50°C for 3.5 hours. Saturated ammonium chloride (1 ml) was added and the solvents evaporated. The residue was chromatographed on silica using chloroform- methanol; yield: 0.0145 g (50%) as an oil.

^■'"H NMR(CDC1₃, 300 MHz)d : 1.24(S,9H) ,1.80-2.00 (m,2H) ,2.10- 2.25(m,lH) ,2.42-2.60 (m,lH) ,3.73-4.12(ABX,2H) ,4.22- 4.34 (m,lH) ,5.23 (s,2H) ,5.27-5.43 (AB,2H) ,6.07(dd,lH) ,7.15- 7.30 (b,lH) ,7.30-7.48 (m,6H) ,8.32(d,lH) .

Example 11

5'-0-Pivaloyloxymethyl-2' ,3'-dideoxycytosine

4 N -Benzyloxycarbonyl-5'-O-pivaloyloxymethyl-2' ,3'- dideoxycytosine (0.05 mmol) is added to a suspension of 5% Pd on charcoal (5 g) in ethanol (2 ml) .

The stirred suspension is subjected to hydrogenolysis at atmospheric pressure using a Brown apparatus where the H₂~ gas is generated in a controlled manner by the addition of 3N HC1 to a solution of sodium borohydride in a separate compartment.

The reaction is run for 1 hour at room temperature, the mixture filtered through a thin bed of Celite, the filtrate evaporated and the product purified by chromatography on a silica gel column using

CHCl₃:EtOH(9:l) for elution.

Example 12

3-Pivaloyloxymethyl-5 '-O-propionyl-3'-azido-3 '- doexythymidine

3 '-azido-3'-deoxythymidine (0.0504g, 0.189 mmol) and 4-dimethylaminopyridine (0.0023g, 0.019 mmol) were dissolved in pyridine (2ml) and cooled to 0°C. Propionic anhydride (0.0265 ml, 0.206 mmol) was added and the mixture stirred at room temperature under nitrogen for 24 hours before the solvent was evaporated under reduced pressure. Toluene was added and evaporated under reduced pressure and the residue chromatographed on silica using chloroform and chloroform-methanol (98.2); yield

0.060 g, (98%) of 5'-O-propionyl-3 '-azido-3 '-deoxythymidin as a colourless oil.

5 '-O-propionyl-3 '-azido-3'-deoxythymidine (0.042 g, 0.130 mmol) and potassium carbonate (0.0198g,

0.143 mmol) were suspended in DMF (1ml) and stirred at room temperature under nitrogen for

1.5 hours. The mixture was cooled to °C and chloromethyl pivalate (0.0226 ml, 0.156 mmol) added. The mixture was stirred at 0°C for 30 minutes and then at room temperature for 19 hours before the solvent was evaporated under reduced pressure. The residue was chromatographed on silica using ethyl acetate- hexane (4:6); yield 0.045g (81%) as a colourless oil.

1 H NMR (CDC1₃, 300 MHz) C : 1.19 (s) and 1.19(t)

[12H], 1.96 (d,3H), 2.29-2.57 ( ) and 2.41 (q)

[4H], 4.06-4.15 (m,lH) , 4.16-4.25 (m,lH), 4.31-4.42 (ABX,

2H) , 5.91-5.98 (AB,2H), 6.15 (t,lH), 7.27 (d,H-6).

¹³C NMR (CDC1₃)S : 9.03, 13.27, 27.02, 27.43, 37.73,

38.84, 60.54, 63.19, 64.95, 81.91, 86.10, 110.49,

134.10, 150.04, 162.27, 173.72, 177.48.

Example 13

3- -(Ethyloxycarbonyloxy)ethyl-5 '-O-propionyl-3'- deoxythymidine

3 '-Deoxythymidine (0.0352g, 0.156 mmol) and 4-dimethylamin (0.0020 g, 0.016 mmol) were dissolved in pyridine (3ml) and cooled to 0°C. Propionic anhydride (0.0241 ml, 0.187 mmol) was added and the mixture stirred at room temperature under nitrogen for 24 hours before the solvent was evaporated under reduced pressure. Toluene was added and evaporated under reduced pressure and the residue chromatographed on silica using chloroform and chloroform-methanol (98,2) ; yield 0.0435 g (99%) of 5 '-O-propionyl- 3 '-deoxythymidine as a semi-solid.

5'-O-propionyl-3 '-deoxythymidine (0.063 g, 0.223 mmol) and potassium carbonate (0.0339 g, 0.245 mmol) were suspended in DMF (2ml) and stirred at room temperature under nitrogen for 1.5 hours.

The mixture was cooled to 0°C and 1-chloroethylethylcarbon

(0.039 ml, 0.291 mmol) was added. The mixture was stirred at 0°C for 30 minutes and at room temperature for 2 hours. The temperature was increased to

60^CC and the mixture stirred for 19.5 hours before the solvent was evaporated under reduced pressure.

The residue was chromatographed on silica using ethyl acetate-hexane (50:50) (R 0.17) ; yield 0.0477 g (54%) as an oil.

^ NMR (CDC1₃, 300 MHz) O : 1.18 (t,3H) , 1.29 (t,3H) ,

1.70-1.87 (m) and 1.86 (d) [4H], 1.94 (d,3H) , 1.97-

2.14 (m,2H), 2.35-2.52 (m) and 2.40 (q) [3H], 4.12-

4.25 (m,2H), 4.26-4.40 (m,3H) , 6.07-6.12 (m,lH) ,

7.20-7.28 (m,lH), 7.36 (d, 1H) .

¹³C NMR (CDC1₃)S:9.09, 13.23 and 13.36*, 14.15,

17.86 and 17794*, 25.81 and 25.84*, 27.48, 32.25 and 32.35*, 64.34, 64.79, 77.49, 78.46 and 78.56*,

86.52 and 86.58*, 109.81 and 109.84*, 133.92 and 133.97*,

149.82, 153.81 and 153.85*, 162.57 and 162.59*,

174.06.

* Different shifts for the two enantiomers (due to )-CH(CH₃)-N) are observed. Example 14

5 '-O-Propionyl-3-α-(thienyloxycarbonyloxy)ethyl-

3'-azido-3'-deoxythymidine

5'-O-Propionyl-3'-azido-3'-deoxythymidine (0.051 g, 0.158 mmol) and potassium carbonate (0.024 g, 0.174 mmol) were suspended in DMF and stirred at room temperature under nitrogen for 1 hour. The mixture was cooled to 0°C and 1-chloroethyl thienyl carbonate (0.046 g, 0.208 mmol) added. The mixture was stirred at room temperature for 17 hours and then at 55°C for 9 hours. The solvent was evaporated under reduced pressure and the residue chromatographed on silica using chloroform and chloroform - methanol (99:1). The product containing fractions, R_f 0.3 (chloroform - methanol (99:1) were composed of two diastereoisomeric components, R_f0.18 and R_f0.22 (ethyl acetate - hexane (4:6); yield 0.050 g.

Example 15 —Benzyloxycarbonyl-5 '-O-pivaloyloxymethyl-2' ,3'- dideoxyadenosine

2' ,3 '-Dideoxyadenosine (0.1 mmol) and 4-dimethylaminopyrid

(0.1 mmol) were dissolved in pyridine and the mixture cooled to 0°C. Benzyl chloroformate (0.2 mmol) was added and the resulting mixture stirred at room temperature under nitrogen for 24 hours.

4-Dimethylaminopyridine (0.1 mmol) was added and the mixture cooled to 0°C. Benzyl chloroformate

(0.2 mmol) was added and the mixture stirred at room temperature for 24 hours. The addition of

4-dimethylaminopyridine and benzyl chloroformate was repeated 3 times with 24 hours of stirring between each addition before the solvent was evaporated under reduced pressure. The residue was chromatographed on silica using chloroform, chloroform-methanol (99:1) and chloroform-methanol (9:1) yielding N - benzyloxycarbonyl-2' ,3 ^r-dideoxyadenosine. N -benzyloxycarbonyl-2' ,3'-dideoxyadenosine (0.1 mmol) and sodium hydride (80% in oil, 0.21 mmol) in DMF (2 ml) was stirred at 0°C for 1 hour. The mixture was cooled to -50°C and chloromethylpivalate (0.1 mmol) added. The mixture was stirred for 4 hours before acetic acid (0.1 mmol) was added. The solvent was evaporated under reduced pressure and the residue chromatographed on silica using chloroform and chloroform-methanol (99:1), yielding the title compound.

Pharmaceutical Example A Preparation of capsules for oral use

Active Compound 50 mg

Amylum maydis q.s.

The powder is mixed and filled into hard gelatin capsules (Capsugel Size 00) .

Pharamceutical Example B Preparation of an ointment

Active compound 1 g

Liquid paraffin 100 g

White soft paraffin to 1000 g

White soft paraffin is melted and incorporated into the liquid paraffin and stirred until the mixture is cold. Active compound is triturated with a portion of the basis and gradually the remainder of the basis was incorporated. The ointment is filled into lacquered aluminium tubes (20 g) and sealed. The ointment contains 0.1 % active compound. - 23 -

Pharmaceutical Example C

Suspension for parenteral administration

Active Compound 200 gram

Polysorbate 80 3 gram

Sorbitol 400 gram

Benzyl alcohol 8 gram

Water ad 1000 ml 1M HC1 q.s.

Polysorbate 80, Sorbitol and benzyl alcohol are dissolved in 500 ml distilled water. Active compound is screened through a 0.15 mm sieve and dispersed in the solution under vigorous stirring. The pH is adjusted to 4.5 by dropwise addition of 1M HC1. Water is added to 1000 ml, the suspension was filled in 1 ml vials The vials are sterilized by y -radiation. Each vial contains 200 mg active compound.

Pharmaceutical Example D Preparation of tablets

Gram

Active Compound 200

Lactose 85

Polyvinylpyrrolidone 5

Starch 42

Talcum powder _ 15

Magnesium stearate 3

Active compound and lactose are screened through a 0.15 mm sieve and mixed together for 10 minutes. The mixed powder is wetted with an aqueous solution of polyvinyl-pyrrolidone. The mass is granulated, and the dried (40 °C) granulate is mixed with starch, talcum powder and magnesium stearate. The granulate is compressed into tablets. The tablet diameter is 11 mm, the tablet weight is 350 mg and each tablet contains 200 mg active compound. Pharmaceutical Example E

Preparation of a suspension for rectal administration

Methyl £-hydroxybenzoate (70 mg) and propyl £-hydroxy- benzoate (15 mg) are dissolved in water (100 ml) at 90 °C. After cooling to 30 °C methyl cellulose (2g) is added and the mixture is agitated for 3 hours. 1 gram active compound are screened through a 0.15 mm sieve, and dispersed in the solution under vigorous stirring. The suspension is filled in a 100 ml tube. The suspension contain 10 mg active compound/ml.

Pharmaceutical Example F Preparation of oral suspension

Gram Active Compound 10

Carboxymethyl cellulose 1.5

Sorbitol 200 Sodium benzoate 1.0

Orange essence 0.3

Apricot essence 0.7

Ethanol 50

Water 236.5

Carboxymethyl cellulose, sorbitol and sodium benzoate are dissolved in water with stirring _for 2 hours. A solution of the essences in ethanol is added. Active compound is screened through a 0.15 mm sieve and dispersed in the solution under vigorous stirring. The suspension (10 gram) is filled in a 20 ml tube. Each tube contains 200 mg active compound. Pharmaceutical Example G Preparation of injection solution

10 mg active compound are dissolved in 10 ml 0.9 % sodium chloride. pH is adjusted to 4.5 with IN HC1. The solution is sterile filtered and filled into a 10 ml vial. The solution contains 1 mg active compound/ml.

Pharmaceutical Example H

Preparation of tablets (controlled release formulation)

Gram Active Compound 500

Hydroxypropylmethylcellulose 120

(Methocel K15)

Lactose 45

Povidone 30

Magnesium stearate 5

Active compound hydroxypropyl-methylcellulose and lactose are mixed together for 20 minutes and granulated with a solution of povidone. Magnesium stearate is added and the mixture is compressed into tablets.

The tablet diameter is 13 mm, the tablet weight is 700 mg and each tablet contains 500 mg active compoun .

Claims

- 26 -CLAIMS:

1. Compounds of formula (I)

(wherein Z is a hydrogen atom or an azido group, Y is a hydrogen atom or a physiologically acceptable group of the formula R¹(0)_nCO(OCR²R³) -

where n is 0 or 1, m is 0 or 1 and

R is an optionally substituted alkyl or aryl group or, where n is 0, a hydrogen atom.

R 2 and R 3 are independently hydrogen atoms or lower alkyl groups; and

X is a group selected from

X (F)

(where the groups Y 2, Y3 and Y4 are as defined for Y and may be the same as or different from

Y 1 or each other, R4 is a hydrogen atom or a group -NY 3Y4, where Y3 and Y4 have the above meanings

5 and R is a hydrogen atom or a lower alkyl group, with the proviso that at least one of the groups

Y , Y², Y³ and Y is other than hydrogen, and that

2 when Y is absent or is a hydrogen atom and any groups Y 1, Y3, and Y4 which are present are either hydrogen atoms or groups R 1(0)nCO(OCR2R3)m where m is 0 then Z is an azido group) and/or salts thereof,

Compounds of formula (I) as claimed in claim

1 wherreeiinn RR iiss sseelleecctteedd ffrroomm ooppttiioonnaallllyy substituted ^Cl-20 ^alk^¹ groups and C_g_₂₀ aryl groups^,

3. Compounds of formula (I) as claimed in claim

1 wherein m represents 1 in at least one of the groups Y 1,Y2,Y3 2 and Y4 R is a hydrogen atom; and R is a hydrogen atom or a methyl group.

4. A pharmaceutical composition comprising as active ingredient one or more compounds of formula (I) as defined in any preceding claim and/or a non-toxic salt thereof, together with a pharmaceutical carrier or excipient.

5. A process for the preparation of a compound of formula (I) as defined in any of claims 1 to

3, which comprises reaction of a compound of formula

(ID

[wherein Y and Z are as hereinbefore defined and

*D

R (0)nCO. (OCR R )m as defined above followed where required by removal of any protecting groups and/or unwanted substituents so introduced.

6. Use of compounds of formula (I) as defined in any of claims 1 to 3, and/or salts thereof, in the manufacture of a medicament for the treatment or prophylaxis of retrovirus infections.

7. A method of treatment or prophylaxis of a retrovirus infection comprising administration to a human or animal subject of an effective amount of a compound of formula (I) or a non-toxic salt thereof as defined in any of claims 1 to 3.

8. A method as claimed in claim 7 wherein the retrovirus infection is an HIV infection.