AU614082B2 - Nucleosides and nucleoside analogues, pharmaceutical composition and processes for the preparation of the compounds - Google Patents

Abstract

A compound of formula (I), wherein the radicals A, X, R<1>, R<2>, and R<3> are defined as follows: A: formula (a) or formula (b); X: (a) O; (b) S; (c) CH2; R<1>: H; alkyl containing 1-3 carbon atoms; -CH=CH2; -CH=CH-CH3; -CH2-CH=CH2; formula (II); -C=CH; R<2>: H; or R<2> constitutes together with R<3> a carbon-carbon bond; R<3>: H; F; Cl; Br; I; N3; CN; C=CH; OH; OCH3; CH2OH; and when R<3> is F; Cl; Br; I; N3; CN; C=CH; OH; OCH3 or CH2OH it may have either the cis-configuration or trans-configuration relative to the hydroxymethyl function at position 4', or R<3> constitutes together with R<2> a carbon-carbon bond, and therapeutically acceptable salts thereof, for use in therapy, in particular for the treatment of HIV virus infections.

Description

AU-AI-16899/88 LNIELETAAROPz RAVZTO PCT W I' ter io aIB eftZAIO INTER(NATIONAL APPLICATION PUl [SHE DEI HE ENT COOPERATION TREATY (PCT) international Patent Clas~sification 4 1 (11) International Peblication Number: WO 88/ 080011 C07D 239/46,405/04, 409/04 (43) International Pubulication Date; 20 October 1988 (20,10,88) (21) International Application Number: PCT/SE88/00l69 (74) Agents: MIKSCHE, Gerhard et al.; AB Astra, Patent and Trademark Department, S-151 85 S~dertfllje (SE), (22) International r~iling Date: 6 April 1988 (06.04.88) (81) Designated States: AT, AT (European patent), AU, 1313, BE (31) Priority Application Numb 8701605-1 (European patent), BG, BJ (OAPI patent), BR, CF (OAPI W\ \A patent), CG (QAPI patent), CH, CR (European patent), CM~ (32) Priority Date: 16 April 1987 (16,04,87) (QAPI patent), DE, DE (Euro~oean patent), DK, Fl, FR (European patel-it), GA (OAPI pa tent), GB, GB (European pa- (33)Pririt Coutry SEtent), HU, IT (European patenlt), JP, KP, KR, LK, LU, 1,U (33 Prorty ounry .~SE(Europ'ean patent), MC, O, NIL (OAPI patent), MvR (GA- "Lnean patent), NO, RO, SD, T )AP[ patent), SU, TD (OA- ~NAM1EDI RE/'TED iL SE]: Bilverstigen 19, S-1$0 23 Enh~mrna LINDBORG, Bjbrn, Gunnar (SE/SE]; Helgaragrtlnd 14, S-1l25 42 Alvsj6 SSE). STENING, Gliran, Bertil Vdirbovflgen 10, S- 51 50 SJderttllje OBERG, Bo, Fredrik (SE/SE]; Askvllgen 27, S-752 52 Uppsala (SE).

A~tYr.P. 8 DEC 1988

AUSTRALIAN

4 NOV 1988 PATENT OFFICE (54)Title:. NUCLEOS IDES Al-tD NUCLEOSI DE ANALOGU-CS, PHARMACEUTICAL COMPOS ITI )N AND PRO- CESSES FOR THE PREPARATION OF THE COMPOUNDS Hi 0 N 0

NH

2

N

o -C CH3 C-CH (57) Abstract A compound of formula wherein the radicals A, X, Rl, R 2 and R 3 are defined as follows: At formula or formula 0; RI: H,1 alkyl containing 1.3 carbon Mo~ms', -CH-- 2

-CH-CH.CH

3

-CH

2 CICHI- formula 1( 2 H; or R! constitute$ together with R 3 a carbon-carbon bo;nd:, R3-, 5- CI: .Br; I-,

N

3 CN; CaCH; OH,, OCH-I; CH,1 2 0H; and when R 3 Is, CI' Br;, I; N 3 CN; C=CH; 01-I:OCH 3 or CI-lOH it may have either the cis-configuratiort or tratis-configuration relative to the hydrocymethyl functiont t nositlon or R 3 0onstitutes together with R! a carbon-carbon bond, and therapeutically acteptabie salts thereof, for use in thetapy,, in particular for the treatment of HIV virus infections.

i A 888-1 88 04 Novel Medicinal Co ounds Field of the invention The present invention relates to the use of chemical compounds and physiologically acceptable salts thereof for the therapeutic and prophylactic control and treatment of the Acquired Immuno Deficiency Syndrome (AIDS), infections by Human Immunodeficiency Virus, hepatitis B virus infections and retrovirus infections and method for such control and treatment in animal and man.

Background of the invention 00 o 00 *0 0 00* ::15 *0 2.00

S*

o

I

00 In the late seventies a new disease was reported, which subsequently was referred to as Acquired Immuno Deficiency Syndrome (AIDS). It is now generally accepted that a retrovirus referred to as HIV (Human Immunodeficiency Virus), formerly known as Human T-cell Lymphotropic Virus (HTLV-III) or Lymphadenopathy Associated Virus (LAV) plays an essential role in the etiology of AIDS.

AIDS is charact.,rized by a profound immunodeficiency due to low numbers -of a subset of lymphocyte-T-helper cells, which are one target for HIV infection. The profound immunodeficiency in AIDS patients makes these patients highly susceptible to a variety of opportunistic infections of bacterial, fungal, protozoal or viral etiology. The etiological agents among viral opportunistic infections are often found in the herpes virus group, Herpes simplex virus (HSV), Variceila Zoster virus (VZV), Epstein-Barr virus (EBV) and, especially, cytomcgalovirus (CMV). Other retroviruses affecting humans are HTLV-I and II and examples of retroviruses affecting animals are feline leukemia virus and equine infectios anaemia virus.

Hepatitis B virus infections cause severe disease such as acute hepatitis, chronic hepatitis, fulminant hepatitis in a considerable number of persons. It is estimated that there are 200 million patients with Schronic hepatitis B infection in the world. A considerable number of the

U

2 chronic cases progress to liver cirrosis and liver tumours. In some cases the hepatitis infections also take a rapid and severe course as in fulminant B hepatitis with about 90 mortality. At present ther', is no known effective treatment against hepatitis B infections.

General outline of the invention A great number of nucleoside analogues -xhibit several antimetabolic activities. They do so by substitutin for or competing with the naturally occuring nucleosides. Recently some nucleoside analogues have been described, which inhibit in cell culture the multiplication of human imnunodeficiency virus (HIV, also called HTLV-III, LAV), the causative agent of AIDS and AIDS-related complex (ARC). Such compounds are for example azidothymidine, dideoxycytidine and dideoxyadenosine. These and ':75 other described HIV-antimetabolic nucleoside analogues have the same geometric relationship between the nucleoside base and the glycosidic S part as the naturally occuring nucleosides, i.e. they are 8-anoners.

We have now, surprisingly, found that some nucleosides and nucleoside

V

l analogues with the opposite geometric configuration, C-anomers, are potent inhibitors of HIV multiplication but not of cell-division.

Anti-HIV activities are displayed by such geometric isomers which have been modified either in the nucleoside base part, the glycoside part or in both parts. The structures of these compounds are disclosed in this 25 invention.

Prior Art The following compounds of the formula I below are known: 1. Compounds of the formula 0 AN

R

1 Rl

HN

0 Ll wher R i ein R 3 is OH and Rlis as follows: s H and CH 3 R is C 2 HS R is nC, T. Nishimura, B. Shin'izu, I. Iwai Chem. Pharm. Bull. (Tokyo) 12 (1964), 147] M. Swierkowski, D. Shugar J. Med. Chem. 12 (1969), 533 A. Szaboles, J. S6gi, L. 6tvbs J. Carbohydrates, Nucleosides, Nucleot~des 2 (1975), 197 211 M. Draminski, A. Zgit-Wroblewska Polish J. Chemistry 54 (1980), 1085 P.J. Barr, A.S. Jones, P. Serafinowski, R. Walker J. Chem. Soc. Perkin I (1978), 1263 1267

N

3 and Rl is CH 3 M. Imezawa, F. Eckstein, J. Org.

3044- 3048.

R Iis i-C 3

H

7 .s

S

S. S S

S.

S

R is C--CH: and wherein R 3 is Chem. 43 (1978), 2. The compound of the formula

S

S S 9* 0

S.

S S

S

S

*SSSS*

S

R

1 is C~CH is described R. Walker, J. Chern. Soc.

O

N

OH I by P.J. Barr, Jones, P, Serafinowski, Perkin 1 (1978), 1263 1267 i R is H is described by J.J. Fox, Yung, I. Wempen and M. Hoffer, J. An. Chem. Soc., Vol. 83 (1961), 4066-4070.

Both groups 1. and 2. concern only compounds having the 3'group and the 4'hydroxymethyl group in a trans-configuration.

Disclosure of the invention Specifically, we have now fjund according to the capounds of the forula: the present invention that

R

3

R

2

JA

2' HO- X c--ancmer and therapeutically acceptable salts thereof, wherein the radicals A, X,

R

2 and R are defined as follows: 9*: 0.00 0 00 0

I

0. V 1 0 a a a.I *5 a A: (a)

HN

N

NH

2 N 1

NR

S wherein R is H7 alkyl containing 1-3 carton atons; 21~ ii;cI 9 -ic 2 C"i 3 ;or -I!U X: 0 or

CH,

r! 1i i: R2: H; and R3: H; F; Cl; CN;-CCH; OCH3; or CH 2 OH, wherein when R 3 in formula I is F; Cl; CN; C=CH; OCH 3 or CH20H it may have either cis-configuration or trans-configuration relative to the hydroxymethyl function at position with the proviso that when X is 0, then R 3 is not H, ,inhibit the multiplication of human immunodeficiency virus (HIV).

The compounds of the formula I are useful as therapeutic and/or prophylactic agents in the control and treatment of HIV virus infections in mammals and man.

In a more general aspect, the compounds of the formula I are useful as therapeutic and/or prophylactic agents in the control and treatment ol infections caused by retroviruses and hepatitis B virus in mammals and man.

All retroviruses, including HIV, require the enzyme reverse transcriptase in their natural cycle of replication.

Hepatitis B virus (HBV) is a DNA virus with a unique circular doublestranded DNA genome which is partly single-stranded. It contains a specific DNA polymerase required for viral replication. This DNA polymerase also acts as a reverse transcriptase during the replication of HBV DNA via an RNA intermediate.

The compounds of the formula I inhibit the activity of reverse tran- S scriptase of retroviruses including HIV as well as the activity of DNA polymerase of hepatitis B virus.

S

The present invention has several aspects: 3 1. the novel compounds included in the formula I, it ai t- L- i Ij 11 r,

I

2. pharmaceutical compositions comprising a compound of the formula I as active ingredient, 3. a compound of the formula I for use in therapy, 4. a compound of the formula I for use in the manufacture of a medicament for therapeutic and/or prophylactic treatment of infections caused by a retrovirus, including HIV, or by hepatitis B virus, a method for the therapeutic and/or prophylactic treatment of infections in mammals and man caused by retrovirus including HIV or hepatitis B virus, by administering to a host in need of such treatment an efficient amount of a compound of the formula I.

It is a preferred aspect of the invention to combat HIV virus infections in man.

The expression "alkyl containing 1-3 carbon atoms" for the radical R means CH 3

C

2

H

5

CH

2

CH

2

CH

3

CH(CH

3 2 and cyclopropyl.

When R 3 in formula I is F, C1, CN, -OCH or CHOH it may have either cis-configuration or trans-configuration relative to the hydroxymethyl function at position 4'.

Preferred compounds of the formula I are: 0

SR

1

HN

A is 0 N

I

or

NH

2 N0 A is 0 I J I _i ,i 7

R

3 at position 3' and the hydroxyrnethyJ. group at position 41 have the trans-configuration R' is C 3or C X is 0 (if) R3 is i-i, F, OCH3, or Cl1 2 OHi e~ C C m~ C .me.

C C e C CC me C 9* C ~Ce.

C

(g R 3 i's H or FI the combination of and above the combination of and (if) above

C

em sCt C .me, C CC me C em C C I'm..

C$ ml C C e e Cem CC C

C

the combination of (if) and above the capbinatiox and above the canbination and (if) above (mn) the caribination I I (if) and above 8 1 Examples of preferred compounds-are: R 3 R 9, 9 .9.

9 49eS* CH 3 CH 3

CH

3

CH

3 C 2

H

5 C 2

H

5 C 2

H

5 4.9 9.

9. 9

'C'

9.

9 9 a 99

H-

OCH

3 CH H 2

OH

F

OCH

3 C H 2 0OH

F

HO-

99 *9 9 9 9 9 J A I 1 9

R

2 is H; R 3 is H

R

2 is H; R 3 is OCH 3

R

2 is H; R 3 is CH 2 0H

R

2 is H; R 3 is F In all the examples of preferred compounds R 3 at position 3' and hydroxymethyl at position 4' have the trans-configuration.

1 0* r 4

S

In clinical practice the nucleosides of the formula I will normally be administered orally, by injection or by infusion in the form of a pharmaceutical preparation comprising the active ingredient in the form of the original compound or optionally in the form of a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable carrier which may be a solid, semi-solid or liquid diluent or an ingestible capsule. The compound may also be used without carrier material. As examples of pharmaceutical preparations may be mentioned tablets, dragges, capsules, granulates, suspensions, elixirs, syrups, solutions etc. Usually the active substance will comprise betwet. 0.05 and 20 for preparations intended for injection and between 10 and 90 X for preparations intended for oral administration.

a 0 S In the treatment of patients suffering from retrovirus, especially HIV, 3. or hepatitis B virus infections, it will be preferred to administer the compounds by any suitable route including the oral, parenteral, rectal, nasal, topical and vaginal route. The parenteral route includes subcutaneous, intramuscular, intravenous and sublingual administration. The topical route includes buccal and sublingual administration. The dosage at which the active ingredients are administered may vary within a wide range and will depend on various factors such as the se r y of the infection, the age of patient etc., and may have to be individually adjusted. As a possible range for the amount of the compounds of the invention or a physiologically acceptable salt thereof to be administered per day may be mentioned from about 10 mg to about 10 000 mg, pre-

A

ferentially 100 500 mg for intravenous administration and preferentially 100 3000 mg for oral administration.

Examples of pharmaceutically acceptable salts of the compounds of formula I include base salts, e.g. derived from an appropriate base, such as alkali metal sodium), alkaline earth metal nlagnesium) salts, ammonium and NX+ (wherein X is C1-4 alkyl). Physiologically acceptable salts of a hydrogen atom or an amino group include salts of organic carboxylic acids such as acetic, lactic, gluconic, citric, tartaric, maleic, malic, panthothenic, isethionic, succinic, oxalic, lacto- .1Q bionic and succinic acids; organic sulfonic acids such as methanesulfonic, ethanesulfonic, benzenesulfonic, p-chlorobenzenesulphonic and S p-toluenesulfonic acids and inorganic acids such as hydrochloric, hydroiodic, sulfuric, phosphoric and sulfariic acids. Physiologically acceptable salts of a compound of an hydroxy grotp include the anion of said compound in combination with a suitable cation such as Na NH4, and NX+ (wherein X is a CI-4 iakyl group), Those compounds of the formula I which are novel are summarized as compounds of the formula I with the proviso that when X is 0, then R is not H.

S 6* r

S

The administered compounds may also be used in therapy in conjunction with other medicaments such as 9- (2-hydroxy-l-(hydroxymethyl)ethoxy)methyl]guanine, 9-(2-hydroxyethoxymethyl)guanine (acyclovir), 2-ariiino- 9-(2-hydroxyethoxymethyl)purine, interferon, OC-interferon, interleukin II, and phosphonoformate, or in conjunction with immune modulating therapy including bone narrow or lymphocyte transplants cations such as leyamisol or thymosin which would incr"ease lyr,.

numbers and/or function as is appropriate.

Methods of pyeparation The compounds of the invention may be prepared by one of the following general methods, constituting a further aspect of the invention, A. Condensing a glycoside as comprised in formula I, where the hydroxyl groups may be optionally protected, to the N-1 position of a pyrimidine derivative, corresponding to radical A in formula I according to known methods described in the literature, followed by separation of the c-anwner andl g-ancmer and remioval of any protectinq group Such me~thods are *Dee 0.0 described for example in "Basic Principles in Nucleic Acid Chemistry", Do :6 Do Vul, 1 (Academic Press, 1974s Ed. P*0,P.Tslo), in "Nucleoside Analogues, ~**.Chemistry, Biology and Medical Applicatfons" (Pharma Press, 1979, adso R.T. Walker, E. De Clercq arnd F. Eckstein) and in Nucleic Acids kesearch Vol. 12, 1984, pages 68?7 6837 Hubbard, A,S, Jones and R.T. Walker)., An example of such a method is given for the case of a uracil base analogue,, R 5 0S0l$(CH3)

HL.

0, -9Hal N

I

2+ 0 2 R R (CH )Sl0 R 4

R

3 3

R

wK~ein R 4 is H, F, Cl, cN, c- cii, Qci$.J or C14 2 OR P iL a pro*fcting group, of which a great variety is known, and examples of which are p-toluoyl, acetyl, trityl, benzyl. R 1 andR are as defined .el above.

12 B. Anomierization of a A-anomer, of the formula

A

X

R 0

RR

wherein A, X, and R2are as defined above, R 4 is H; F; Cl; cw;

OCH

3 or CH OR5; wtierein R5 is H or a hydroxy-protecting group to a midxture of o and 0-=ner.s, ,hereaftler the ct-anomer and -anc~ier are separated and any protecting groups removed. The anonlerization may be performed by known methods, e~g, with an optionally protected P-nucleoside, for example a silylated nucleoside, wi-,a a cata~lyst, such as for example trimethylsilyl trifluoromethanesulfonate OSime 3 OSiMe 3 4.3 R R 0 o .R I 1 R 2 R 4 and R 5 are as de~ined above.

C. A transglycosylation reaction whereby the sugar mnoiety forming a bond, c4.- or -,to one nucleoside base, is transferred to the desireQU pyrimidline base. The reactio n is performed with a catalyst such as for is example trimothylsilyl trifluoromethanesulfonate, and is Jfollowed by separation of the products ane doprotectiog.

OSi (CH 3 3 0

I

Si (CH 3 3 OSi (CH 3 3 0 NN other products 0 00 so so wherein R 1

R?,

midine or purin Rand R5are z.s defined above. The radical B is a pyribase, the choice of which is not critic;al, D. Introductiojn of the functional group R 3 or a precursor of R 3 into the nvcleoside c<-anomer by substitution of a suitable leaving group,R7 followed by deprotection.

AN

R

R *0 o~ N

A

4

I

R and R are as defined above, R 7 is a good leaving group such as for 8 example trifluoromethanesulfonyloxy, R is F, cl, CN,

OCH

3 and synthos for the C=CH, and CH20H groups, such as for example C=C-Si(CH 3 3

CH

3

CO

2 and HC"S R is a suitable protecting group.

An alternative way for introduction of the R 8 function is by reaction of the 2,3'-anhydrocK-anomer.

0

R

50 0

HN

0 N 8 5 R 0- 0 0

S

S.:

*5 S S

OSS

0

*SSS*@

wheei 1I R5 are as defined above wherein R R and R are as defined above.

I

The principles of methods A-D above both uridine and cytidine analogues form'las illustrating the reactions are applicable to the synthesis of of formulas I and II, although the only depict uridine analogues.

0St S S

OS

SS 00 S S.

S S E. Converting the uracil moiety of the 5-substituted or unsubstituted CC-uridine compounds to a cytosine moiety of the corresponding GC-cytidine analogues. This is carried out by conventional methods, the principles of which have been described for example by WL. Sung Chem.

Soc. Chem. Commun. 1981, p. 1089 and J. Organic Chemistry 1982, volume 47, pages 3623 3628) ,nd by P. Herdewijn et al. Medicinal Chemistry 1985, volume 28. pages 550 555).

The following examples will further illustrate the invention.

i .1 Preparation of intermediate products A. Preparation of 3{1-3'-deoxy-5'-0-acetylthymidine (VSB423) 3'F-31-deoxythymidine 45 mg (0.184 mrnol) in acetic anhydride (2.0 mL) was heated with stirring in an oil bath at 800 for'7 hrs. The solution was evaporated in vacuo and the residual acetic anhydride and acetic acid were removed by several additions and reevaporations with benzenetoluene The residue was used without further purificati .n(s.

Preparation of com- ounds of the invention

S*

S.

0@ p 005S 0

S..

5, esesep p @5 0S 0

SO

S@ S 5550 p. *e *0 0 ~A 0 5 Example 1. Preparation of l-(3-F-2,3-dideoxy-O(-0-ribofuranosyl)thymine (VSA 419) (Method 8) Thymine 23 mg (0.18 mol) and 3 9 F-3'-deoxy-5'-0-acetylthyiidine was suspended in acetonitrile (1.2 mL) and N,0-Bis (trimethylsily1)-acetamide (0.35 mL) was added, The mixture was stirred at room temp~erature for 1.5 hrs. Trimethy'isilyl trifluoromethanesulfonate (0.05 mL) was added. After stirring at room temperature for 192 hrs., the mixture was poured under stirring into a 1:1 mixture of 20 ml of 10 aqueous KHCO 3 ethyl acetate. Two phases were separated and the water phase was extracted with ethyl acetate (3 x 10 mL). The combined ethyl acetate phase was filtered and evaporated in vacuo. The residue was dissolved in dichloromethane-ethyl acetate 1:1 and applied to a column of zilica gel, and the column was eluated with dichlorcumethane-ethyl acetate 1:1 to give 28 mg (53 starting material (VSB 423) (iRf 0.37 on TLC silicagel

CH

2 ;Cl 2 EtoAc 1:1) and 18 mg (34 of l-(3-F-2,3-dideoxy-5-0-acetyl- -c(-0-ribofuranosyl)thymine (VSB 424) (Rf 0.29 on TLC silica gel CH 2 C1 2 -EtoAc 1 MMR (CD 3 0D) 1.95 3H, CH 3 2.12 3H, CH 3 CO), 2.3-3.0 (in, 2H, H-2 "ab) 4. 15 (dI 2H, J4 I, 51P4.4 Hz, H-51 a,b) 4.81 (dt, 1W, J3 IF,4 1 30.0 Hz, J4 1 5 0.6 Hz, H-4')1 5 .23 (dd, lH, J3 IF,31 =53 .7 Hz, J2 s 6.36 1H, JI1',21:7.57 Hz H-i1), 7.27 lH, J H-6, CH 3 =1.47, H-6) j 16 13 C (CD ON)d(12.80 (CH 3 20.87. (CH CO), 39.40 J=20.8 Hz, C-2') 63.37 J=12.2 Hz C-51), 84.65 J=24.4 Hz, 86 .50 (s C-1' 93.82 J=178 Hz, 111.12 135.07 J=6.1 Hz, C-6), 150.48 163.68 170.30 (CH 3

CO).

The compound VSB 424 (16 mg) was dissolved in saturated methanolic ammronia (5 mW and left at room temiperature overnight. The solution was evaporated and the residue was treated with acetone-benzene to .1 give crystals of the desired compound, VSA 419 (9.4 mg, 69 UV A max (H 0) 269 nm.

NMR (DMSO-d6) 'H l1.79 3H, J CH 3 1 H-6-.2 Hz CH 3 2.16-2. 2H, H-21), 3.2-3.6 Cm, 2H, H-51), 4,61 (dt, 1H, j3'F,4'=23.4 Hz, J4',51= Hz, H-41), 5.06 1H, Hz, OH), 5.32 (dd, 1H, J3'F,3'=54.2 Hz J2'31=4.9 Hz, 6.18 (dd, 1H, J1'2'=7.7 Hz and 2.1 Hz, 7.39 1H, J CH 3 H-6=1.2 Hz, H-6) 13*. C3 (DMSO-d6) d12.46 (CH 3 61.17 J=11.0 Hz, C-51), 85,85 87.15 J=20.8 Hz, C-41), 94.75 J=173 Hz, C-31), 109.15 135.63 J=6.1 Hz, 150.53 163.95 (C-4) Example 2. Preparation of l-C3-F-2,3-dideoxy-ci.-D-ribofuranosyl)- (VSA 409) (Method C).

:n 5-Propyluracil (56 mng) and 3'-F-31deoxy-thymidine (47 mg) were suspended in acetonitrile (1.2 mL) and N,0-Bis (trimethylsilyl) acetaniide (0.35 mL) was added. The mixture was stirred at room temperature for hrs. Trimethylsilyl trifluoroinethanesulfonate (0.05 mL) was added.

After stirring at room temiperature for 138 hrs, the mixture was evaporated in vacuo adadded to H 0 (0.5 Wl., filtered and washed with H 2 0 6:060: (0.5 ,10, The combined water phase was applied to a C 18 column (HPLC) and eluted with (iethanol -water (35:65), at a rate of 7.0 mi/mmn. The /3-anonier eluted after 12.9 min, and the desired C-anomer, VSA 409, after 18.0 min. Yield 9.3 mg 18 UV Amax (H 2 0) 269 nin, MS M1 272 10 154 100 119 (76 1 A 17 Example 3. Preparation of l-(3-F-2,3-dideoxy-O-D-ribofuranosyl)- (VSA 411) (Method C) (51 mg) and 3'F-31-deoxythymidine (48 mg) were suspended Mn acetonitrile (1.2 mL and N,0-Bis (trimethylsilyl) acetamide (0.35 WL was added. The mixture was stirred at room temperature for hrs. Trimethylsilyl trifluoromethanesvlfonate (0.05 mL) was added.

After stirrinq at room temperature for 161 hrs, the mixture was evaporated in vacuo, and adddt! to water (03~ mL), filtered and washed with water (0.5 mL) The combined water phase was applied to a C 8 column i0 (HPLC) and eluted with methanol-,,a qr at a rate of 8.0 mi/mmn. The 3 -anomer eluted after 12.3 mm innd the desired X-anomer, VSA 411, after 16.4 min. Yield 13.1 mg (26 ).UV A max (H 2 0) 267.5 nm. MS M+ 258 140 (100 %,119 (67%) ~.Example 4. PreparatIon of l-(2-deoxy-OC-0-ribofuranosyl)-5-isopropenylqs1L5 uracil MVA 115) (Method A) *91SOO: 5-izopropenyluracil (4.3 hexamethyldisilazane (50 ml), chlorotrimethylsilane (1 ml) and ammoniumsulfate (catalytic amount) were heated at reflu.x for 2.5 nrs. Excess of solvent was evaporated in vacua and the residual bis-silylated 5-isopropenyluracil (8.4 g) was dissolved Q in dichioroethane (50 ml) and added to 2-deoxy-3,5-di-0-p-toluoyl- -erythro-pentosyl chloride (11.0 g) in dichloroethane (150 nil) also containinq molecular sieves (4 A, 15 The suspension was stirred at room temperature overnight, after which it was filtered and the solvent was evaporated. The resiOdue was rkAdissolved in dichloromethane which was of 025 washed with saturated aq NaHCO 3 an~d H 0, dried over Na SO 4 and concen- 0 trated to a volume of about 70 nil.- A precipitate foroned which was filtered off, dichloromethane was evaporated from the filtrate and the ,residue was subjectedi to chromatography on silica gel columins eluted with hexane/ethyla, etate/dichloromethane to give l-(2-deoxy- 3 ,5-di-O-p-toluo~yl-G(-D-ribofuranosyl)-5-isopropenyluraciI (VSA 174), 2.64 g (Thin lay ,r chromatography, silica gel, solvent system as above, I c I I I Mwi4- Sodium metal (0.25 g) was dissolved in dry methanol (263 ml), compound VSA 174 (2.'64 g) was added and the solution was stirred at room temperature overnight, after which water (35 ml) was added. The solution was neutralized with an ion exchanger (Dowex H+ 50Wx2), filtered and the solvent was evaporated. The residue was washed with hexane and purified by chromatography on a column of silica RP18 eluted with 50 aq methanol to give 1-(2-deoxy-C(-D-ribofuranosyl)-5-isopropenyluracil. (TLC silica RP8, 50 aq methanol, Compounds of the formula I have been synthesized wherein X is CH 2 and 0, R2 is H and R3 is CH20H; R3 having the trans-configuration relative to the hydroxymethyl function at position These compounds are all novel and have the general formula: qp t

O

0* 0 0v *0 S 9r 0 I S HO

A

HO

X is CH 2 and 0 The synthesis of the compounds where X is CH2 is depicted in the attached scheme. The compounds where X is CH 2 can be resolved into enantiomers by methods for separation of enantiomers known to those skilled in the art. In the examples a detailed description is given for the preparation when RI is H or CH 3 but compounds where Ri is ethyl, propyl, isopropyl, cyclopropyl, ethenyl, prop-l-enyl, prop-2-enyl, isopropenyl and ethynyl, can also be synthesized.

The carbocyclic nucleoside analogs (X CH 2 in the scheme were constructed by forming a pyrimidine ring onto the amino group of ll(3,4trans-C-dibensyloxymethyl)cyclopentyll amine (compound Addition of an isocyanate derivative of alkoxypropenoate, which can be prepared by known methods, followed by acid catalyzed cyclization give the dibensyloxy protected carboxyclic nucleoside analogs 10 pnd 16. Deprotection with trimethylsilyl iodide affords the carbocyclic nucleoside analogs 11 and 17 (Examples 7 and 8, respectively).

The uracil base of compound 10 can be further functionalized by reacting compound 10 under conditions known to those skilled in the art, and described by for example by Bergstrom et al. Am. Chem, Soc. 1976, vol. 98.

19 pp. 1587-1589) and Bigge et al. (Ibid. 1980, vol. 102, pp. 2033-2038). In this way compounds are synthesized wherein RI is ethenyl, propenyl, isopropenyl or ethynyl.

The cyclopentylamine, compound 8, is built from sodium cyanide and diethylmaleate. The formed ketone (compound 1) is protected, the two carboxylic acid ester groups are isomerized to trans, reduced and the formed hydroxyl groups are protected (compound The ketone is selectively deprotected, an oxime is formed and reduced to the amine, compound 8, This amine can be resolved into enantiomers by methods for separation of enantiomers known to those skilled in the art, The carbocyclic nucleoside analogs (X CH 2 where R3 is stereochemically in a cis position relative to the 4' CH20H group have been prepared in a fashion analogous to that described in the attached scheme, but using starting material with the opposite stereo chemistry. In this way the two compounds in Examples 11 and 12 were prepared.

Ais o*o 0 N 2 The trans-3,4-dihydrxyethy compounds of formula I wherein is Example 12 A is Hi CH 3 The trans-3,4dihydrcxymethyl compounds of formula I wherein X is 0, are synthesized by reacting a persilylated pyrimidine base with methyi-(3o@Q 'ohydroxym ethyl)-2,3 -dideoxy-D-erythro- pentofuranoside containing two hydroxyl.protecting groups (benzoyl), The reaction is catalyzed by a silyl triflate ester and give the aQ and the P-ancmers in approximately equal amounts. After condensation the benzoy groups are removed. The VA i i ^y preparation of the a- and P-anomers of the cytosine analog is given in Example 5 and the preparation of the a- and P-anomers of the thymine analog is given in Example 6, Y NH; O YSi(CH 3 3

HO-

Bz OCH3

N

Y A SN HO 0 BzO Si(CH 3 3 The 3-hydroxymethyl pentofuranoside ring is constructed by reacting a mono-protected epoxide of 1,4-butane-diol with allylmagnesium bromide (the scheme is shown in the Experimental part describing the synthesis of the starting materials for Examples 5 and The allyl group is then oxidized with osmium tetraoxide and sodium periodate, cyclized to form the '*fio: pentofuranoside ring and the hydroxyl groups are protected.

b Examole 5. Preoaration of 1-r2'.3'-Dideoxv-3'-C(hvdroxvmethvl)-a-and-O-Dervthro-oentofuranosvll-cvtosine (compounds 21a and 21b) A suspension of cytosine (120 mg, 1.08 mmol) and a small crystal of

(NH

4 2 80 4 in hexamethyldisilazane (2 ml) and trimethylchlorosilane (0.2 ml) was refluxed until a clear solution was obtained. The solution was concentrated in vacuo and co-evaporated with dry xylene. The solid residue was dissolved in dry CH 2 C12 (2 ml) under nitrogen and methyl-5-0-benzoyl-3- [(benzoyloxy)methyl]-2,3-dideoxy-D-erythro-pentofuranoside (170 mg, 0.46 mmol) was added followed by the addition of t-butyldimethylsilyl-triflate (0.22 ml, 0,96 mmol). After 24 h at room temperature the reaction was quenched by the addition of aqueous NaHC03 (sat,) and the resulting mixture was stirred for 30 minutes. The solution was diluted with CH 2 C12 Sand washed with NaHC03 dried, filtered and concentrated to give an anomeric mixture of the protected nucleoside. This mixture was treated with methanolic ammonia (20 ml, sat.) for 24 h at room temperature. After concentration, the residue was dissolved in water and extracted with CHCl.

The aqueous phase was concentrated to a small volume and was applied to a i j- 21 Scheme for synthesis of the compounds wherein X=CH 2 EtO 2 2 EtOt -naCN 002 C0 2 C H 3 HC1 CH 3 OC

HOCH

2

CH

2 0OE

HO

L1A1H 4 NaOCH., 0 99909 9 9 9* 9 9 9 9.

99 9 9 00** 999000 9 9 COOCH 3 3 NaH C H 5

CH

2 B r

EI+

S

9999*9 9 *9 9S 9 *9 *0 9 9990 LIA III

:N-OH

NH

2

,OFI

.0 9 ociHfl

OCH

2 cont I Scheme cont'd Em0

II

0 0Dm R H3 H 400* r 0@ S 0**e 0 0 0@ 0 4R a *a

S.

@0 .040..

U

10 16 R =H R =CH 3 11 R 1

=H

1 17 R C3

NO

2 0 N U 0 0 0* U Re 4 0

SR..

*0 @0 44 6

S

0 d@0005 O 0 Dm0 16 l3tnQ 1 R ~U Cu 3

QBW

13 Rl 1 jU 19 R !C.1 Bm=t -CUl 2 -0 14 ±l

R#C

S SO S S S S. S 5 4* 5 9

S.

0* S S 20

S

S

S* S

S

5.

S a a 3Q 9 semi -preparative C-18 reversed phase chromatography column and eluted with water containing 2 methanol. First the a-isomer was collected1, followed by the P-isomer. The appropriate fractions were combined and evaporated to give 33 mg of the a-anomer (30 and 27 mng of the f-anoiner (24 a-Anoiner: [oi] 26 D: -54' (c 0.3, H 2 UV (H 2 0) Amax: 272 nin (c 10894); 1H..NMR (270 MF-z, 1D20): 1.92 (m,J2'a,2'b 13.5 Hz, J2'a,3' 9 Hz, J2'a,11 Hz, 1H, 2,5 (mn, 1H, 2,7 (in, J2'a,2'b 13,5 Hz, J21a,3' 8H2, J2'a, 1' 6Hz, 111, Ii-2'b); 3.67, 3.69 (d and q, overlapping, J 6 3 1 6.2 Hz, 12.5 Hz, J41,5'a 5.3 Hz, 3H, H-6'and 3.85 J5'a,5vb =12.5 Hz, J4',51b 3 Hz, 1H, 4.28 (in, J 3 4 =8 Hz, J41,5'a 5,3 Hz, 3 Hz, 1H, 6.1 (d and q, overlapping, J 5 6 7.3 Hz, J1',21 6.5 2H, and H-i1); 7.8 (di J5, 6 7.3 Hz, 1H1, 13C-NMR (25.05 MHz, D 2 0): 36,4 42.3 62.7, 63,6 84.4, 88,2 96.6 141.9 158.1 166,8 -Anomer: EQIJ 2 6D: 64'C (c 0.27, H 2 IN (H20) Xmx 272 inn( 9208); 'H-NMR (270 MHz, D 2 0) 8 2,2.-2,46 (mn, 3H, H-2'axnd 3.68 Kd J3,'=5,5 Hz, 2H, 3.76 (dd, J41,5,4 5,5 Hz, J5'a,5'b 12,5 Hz, II, H- 3.92 (dd, J4-,5rb =2,9 Hz, J5' 0 ,5'b =12,5 Hz, ItH, 4.01 (in, J81,4' 8,1 Hz, 5.5 HZ, J5'b,4- 2.9 Hz, IH, Hi4%) 6,05 J6,6 7,3 Hz, 1H, 6,11 (dd, J11,2'a 7.0 Hz, Ji', 2 'b 4,0 liz, IH, 1i,1 7.91 J5,6 7.3 Hz, IH, 13C-.NNMR (25,05 MHz, D 2 0) 8 36,1 40,8 62-7,163,1 and 84.7, 87,1 (C-11 and 96,5 (C45)) 142.2 158.2 166,8 Anal. Oalcd, for C1OHISO04N3xO.7 H 2 0: C, 47.3; H, 16.6, Found 0, 47,2; H, 5.8; N, 16.4.

Example 6. Preparation of 1-r2',3'-Dideoxv-'-C-(hydroxvinethvlb-a-and- ri-Berythro-pentofuranosylI-thivine (compounds 22a and 22b) Thymine (150 mng, 1.19 rnmol) was condensed with methyl 3-Cf(benzoyloxy)inethylJl5O'.-pbromobenzyl-2,3-dideoxy-D-erythropentofuranoside (205 mg, 0.47 mine 1 following the same procedure as described in Examnples 1 and 2 to give an anoineric mixture of the protected nucleoside, The mnixture was dissolved in ethanol containing NaHCQ 3 (excess) and was hydrogenated over Pd (10 on carbonj 1 atnm) for 3 hours, 24 After workup the residue was further reacted with methanolic ammonia for J 24 hors. After concentration the residue was dissolved in water and extracted with 0112012. The aqueous layer was concentrated to a small volume and was subjected to semi-preparative C-18 reversed phase column chromatography and eluted with water containing 10 methanol- First the al-isomer was eluted, followed by the 1-isomer, The appropriate fraction~s were combined and evaporated to give 40 mg of the a-anomer (33 cc) and 41 mg of P-anomer (33 cl-Isomer; 2 6 D: 0.36, 1120); UV (H 2 0) Xmax: 268 nm IQ 13756); 1H-NMR k270 MHz, D 2 0):1,89 J 1.1 Hz, 311, 5-OH 3 1,96 (in, J2'a,2b' 13.2 Hz, J2'a,3' 9,9 Hz, J 2 'a, 1 1 7.7 Hz, 1H1, 2,47 (mn, 1H, H- 2,6 j2'a,2'b' 13,2 Hz, J2%b,31 8,1 Hz, J2'b,1 6,2 Hz, 1H11, H-2'b); 3,64 and 3.68 (d and q overlapping, J 5 ia, 4 t 5,5 Hz, J5'a,5'b =12,1 Hz, J 6 3 6.1 Hz, 3H1, H-5'a, 3.81 J5'b,41 2.9 Hz, J5'b,5'a 12,1 Hz, 4,24 (in, J31, 4 1 8,4 Hz, J41',Sa 5.5 Hz, J 4 5 'b 2.9 Hz, 1H, H1-4); 6,11 (q, J11,2'a 7,7 Hz, J10,2-b =6.2 Hz, 1H1, 7,59 J =1.1 Hz, 111, 13C- NMR (25,05 MHz, D 2 12.5 (5 C'H3); 35,7 42,7 62,6, 63,5 84,2,787,3 111.6 138,1 152.4 167.3 9.13-some: El 2 17,8' (c 0,41, H120); UV (H120) Xm 268 Rim (C 2Q 8516); IH-NMR (27Q MFz, D 2 8 1.91 31I, 5-CHa), 2,3 (mn, 2H,1H-2%) (in, 111, 11-3%) 3.69 J3,, 6 1 5,1~ Hz, 211, H1-6%) 3.76 (dd, J 5 12,4 Hz, J5'b,4' 5,1 Hz, 1H1, iE-5'b), 3,9 (dd, J5'a,5'b =12,4 Hz, 2.9 Hz, 1H1,H 3,99 (in, i4',5'a =2,9 Hz, J4'5'h 5.1 Hz, J 4 1, 3 0 8,1 Hz, 111, 114), 6.14 (dd, Jl', 2 a 4,8 Hz, Jl,2'h 6,8 Hz, 111, 7,73 J -1.1 Hz, 111, 11-6).

l130-NME (25,05 WIZ D20) 8 12.5 (5-0113), 35.4 40.9 62.8, 62,9 0C6), 84.5, 86.1 and 111,7 138, l 152,7 (C-2)1 167.5 Anal, Calcd for C1 111d06N2x0,8 1120: C, 48,8-,11, N, 10.4, Found C, 48.9; 11, N, 10.2.

Preparation f intermediate products 30 The swating naterii for the compounids 21 and 22 i n Examples 0' and 6 respt,ively were prepared by the following sequence of reactions a d- ,61

OR'

OR'

HO0,., 01)

OR'

R '-Brombnzyl

R'-H

a

RS-H

R'sp-Bromobenfl~y Ra BarizoyI *0.0 S S 5* es's S S 6# S 9@ S S

S

6Oeses

S

S

SS

S. S S S 0 R a 00 ,Ii. Rl ,pramobnzyl R Benlzo B' S IR 8.f 4 y

S

S S

S

4) (2S,3I)--P-Bomobenzyt34.*42propeny1 )4 1 4butantriol, To a cold solution of allylmagnesitm bromide .501C (20 mnl, iM) in INO ml of dry diethyl ether under itrogevi atorinosphere, a soltionw c(2S,8R)*34((4bronobenizyl)oxylmetylloxirae2-m'ethano.-i (1,3g, 5 UT IIo In 4 lo dry diethyl ether wvas added dropwise over 30 minutes, The mixtur- Nwas vigorously stirred for 30 minutes at -5QOQ and then quenched wvIth saturaited aqueous ammnonium chloride, The organikc phase was collected '.nd the aqueous phase was extracted wvith diethyl. ether, The organic phases were 9 9 9@ f *9 99 q.)n 9

B

25 9, a *9*9 q 9* .9 6

UO

a 0 *9 09 S 9 *00000 6 combined and washed with hydrogen chloride sodium hydrogen carbonate dried, filtered, concentrated and separated by column chromatography (toluenie: ethyl acetate, to give the title compound (1 if 64 [ojJ 22 D: 1.56' (c 1.03, CHCl3); 111-NMR (100 MHz, CJJ 13): 1.8 (m 1H1, H1-3); 2.13 J 1 1, 2 1 JI3= 6.8 H-z, 211, 3.3 and 3.0 (broad, 211, 011-2); 3,59 (in, 4H1, H-1, 4.0 (mn, 1H1, H41,%; 4.48 2H1, CH 2 Ph); 4,94 and 5.09 (25.05 MHz, C.DC1 3 30,6 42,3 63.3 71.9, 72,3, 72,5

(CH

2 Phi, 04, 116.4 121.5, 129.1, 131.3 (aromatic 136,4 (C-2 and aromatic C); b) (2S,3T?$ 4-0-Benzoyl-1-0-p-broinxobenzyl-3-C-(2'-propen,'l)-1, 2 4 butan-triol. Benzoyl coride (3.21 ml. 27(,6 mmol) was added dropwise to a solution of compound 2 (8,54 g, 27 mniol) in dry pyridine (50 ml) at 0CC. The reaction mixture was stirred at 0 0 C for 15 minutes. Water (5 ml) was added and the solvent was evaporn ted. The residue was dlssolved in dichloromethane and washed with hydrochloric acid (1 M aqueous sodium hydrugen carbonate dried, concentrated and purified by flash chromatography (toluene,, ethyl acetate, to give the title comp~und (9.77 g, 86 [aI 2 2

D:

8,5Q (c 0,71, C11C1 3 IH-NMR (100 MHz, CDCl 3 1.95-2.43 (in, 311, H1-3, H- 2,59 JHj(cO1Th2 3,9 H~z, 1H-, 011-2); 3.54 (in, sec. order, 2H1, H-1.a, LI-ib); 3,95 1H1, 11-2); 4,34 J 4 ,3 5.1 Hz, 2H1, H1-4); 4.48 211, C11 2 1h); 5,14 and 5,0 (in, 2H1, I-3'a, 11-3'b); 5.78 (mn, 111, 11-21); 8,1-7.14 (in, 911, arorn,); 130.

NMR (25,05 MHz, CDCl 3 31.3 40,4 "70.1 0-2; 64,0 (C-4)-,72.3 (CFh); 72.5 116,8 (G-0 1; 121.4-134.7 (aroin.); 136.4 166.1 (COPh).

CY Methyl 3,-1(benzoyloxy)iethiyl]-5-0-p-broinobenzyl-2,,3-dideoxy-Derythro-pentofuranoside, To an ice cold mixture of compound b (7.5 g, 17.9 xirnol) and. N methylinorpholin& N-oxide (4.8 g, 35,5 inmol) in tetrahydrofurantwater (3170 0S04 in t-Butanol (118 ml, 0.02 M, stab. with 1 'PBHP, 0.36 mmzol) was added. After a few minutes, the ice bath was removed and the reaction mixture was stirred overr~ght at room temperature under nitrogen., NaHS03 (2 g) was added and the mixture was stirred for minutes, The solvent was evaporated off and the residue diluted with ethyl acetate, wash.Ad with H10(I NaHCO3 dried, filteredc and concentrated, The crude product was dissolved in te trahydrofuran,.water (3:1, 200 ail) and treated wvIth. NaJO4 (7,65 g, 35.8 intnol). The cis diol was 27 completely cleaved after 30 minutes at room temperature. The tetrahydrofuran was evaporated off and the aqueous residue was saturated with NaC1, and extracted with diethyl ether. The organic phase was dried and concentrated. The residue was treated with methanolic HC1 (0.05 50 ml) for ten minutes, neutralized with Dowex 2x8 (HCO3), filtered, evaporated and the residue was purified by flash chromatography (toluene ethyal acetate, 3:1) to give an anomeric mixture of the title compound (6,63 g, 85 as a colourless syrup. 1H-NMR (100 MHz, CDC13): 1.7-2.9 (three m, 3H, H-3, H-2a, H-2b); 3.31, 3.35 (2s, 3H, OCHs); 3.6 2H, 4.1 1H, 4.4 2H, 4.6 2H, CH2Ph); 5.1 1H, 7.1-8.0 9H, aromatic); 13C-NM. (25.05 MHz, CDCI 3 35.6, 36.4 38.7, 39.3 54,3, 54.5 (OCH3); 65.7, 66.6, 71.5, 72.35, 72,37, 73.8 (CH 2 Ph); 79.9, 80.1 (C- 104,8 121,0-136.4 (aromatic); 165.8 (COPh), d) Methyl 5-0-benzoyl-3-C-[(benzoyloxy)methyl]-2,3 Jideoxy-Derythro-pentafuranoside. A solution of compound r '1 g, 2,3 mmol) in dry diethyl ether (3 ml) was dissolved in liquid ammonia (50 ml) in a dewar bottle. Sodium (300 mg, 13 mmol) was added in portions over 5 minutes. The solution was stirred for 30 minutes and then quenched with solid NH 4 C1, The ammonia was evaporated under a stream of nitrogen and the solid residue S 20 was diluted with ethyl acetate, The solid was filtered off and washed several times with ethyl acetate. The filtrate was concentrated and then coevaporated with dry toluene, The crude residue was dissolved in dry pyridine ml), Benzoylchloride (0.8 ml, 6.9 mmol) was added and the solution was stirred for 40 minutes at room temperature after which water (5 ml) was added and the mixture was concentrated to dryness. The residue was dissolved in CH 2 C1 2 and washed with aqueous HC1 (1 aqueous NaHC0 3 dried, filtered and evaporated to dryness. Flash chromatography (toluene ethyal acetate, 3:1) yielded the title compound (580 mg, 68 as an 3. anomeric mixture, A small sample of the compound was separated by silica 30 gel. 1H-NMR and m.p, of the p-anomer were in agreement with those previously reported, Example 7. Preparation of 14(3,4-trans-Cdihvdroxymethvl)cvclopentylluracil (compound 11) The 3,4-Dibensylcyklaminr (1.0 g, 3.1 mmol) (compound 8) was dissolved in 10 mi C6H6 under nitrogen. C-Ethoxyacryloylisocyanate (Y.F, Shealy, J. Heterocycl. Chem, 1976, V' 1 13, p. 1015) (0.38 M in CH6, 8.2 ml, 3.1 mmnol) was added at 20'C and with vigorous stirring. After a reaction time for 20 minutes the solution was evaporated and the residue was dissolved in trifluoroacetic acid (20 ml, 70 %l aqeous). The solution was refluxed for minutes. the solu'oion was cooled and diluted with H 2 0 (50 ml) and extracted with Et 2 O/Toluene 1:1. The organic phase was washed with H120 (3 x 50 ml) and dried with Na2SO4 anad NaHCO3.

Filtration and evaporation gave a pale brown gum of 1-[(3,4-trans-Cdibensylomethyl)cyclopentyll uracil (compound 10), This gum was dissolved in C11 2 01 2 (5 ml) under nitrogen and was treated with trimethylsilyl iodide (0.72 ml, 5,11 inmol). The resulting cloudy yellow solution was stirred at for 60 minutes. The solution was evaporated, dissolved in CH 2 Cl 2 and extracted with 1120 (3 x 2 ml), The aqeous phase was washed with Et 2 O (3 x Ynl) and evaporated).

Final purification was made by flash-chromatography (Merck S11 elution with etylacetate 7-methanol 1-1-120 0.5. Evaporation of the pure 'S fractions gave the title compound as a colourless gum.

'11-NMR Bruker 250 MHz, D 2 0; 1.5 2.4 ppm-0H2, C11N, 411 0110120,211, 3A4 u.8 ppm (01120,411(m) 4,7 4,9 ppm. CHN, 1, 5.8- 5,9 ppmn (OH =0CH, 1H 7.7 -7.8 ppm CH =CH- 1H-1(d).

2Q Example 8. Pre pa:L atio n of 1- 4- trans- C-di hvdroxvm ethyl-)cvclo.

Pen tyll thymine (compound 17) The compound was prepared by the procedure described for compound 11 in Example 7, substTf tuting inethoxymetacryloyl-isocyanate (prepared according to G, Shaw an" I~N, Warirener, J, Chem. Soc, 1958, p. 157) for ethoxyacryloyl isocyanate.

'a'111 NMR Bruker 250 MHz D 2 0;'1.87 ppm '.C13 1.9-2.3 C1 2 C11N (in) Exarue 9. Preuaration of 14f3,4- trans- C-dili,,Aroxvmeth vl (cvclo- Pentvllcytosine (comoound 14) 1- trans- C-D ib ensyloxyine.-thyl)cyclopentylI uracil1 (compound obtained as described in Examnple 0.5 g, 1,9 numol was dissolved in CH,,H2 ml) and triethylainine (0.31 ml, 3 mnmol), mesitylem~ulfonylchloride (Q.38 17 15 *6*S 0:0 me S

C.

0 4, :2 0 25 9h g, 1.5 mmol) and dimethylaminopyridine (0.037 g, 0.3 mmol) were added under a nitrogen athmosphere and vigorous stirring. The solution was stirred at 40°C for 2 hours and 2-nitrophenol (0.67 g, 5.0 mmol) was added together with triethylamine (0.50 ml, 3.6 mmol). The solution was stirred over night.

Diethylether (50 ml) was added and the solution was washed with 0.5 N NaOH (3 x 20 ml), The organic phase was dried with Na 2 SO4 and evaporated.

The residue was purified by flash-chromatography on silica gel (Merck Si (elution with EtoAc/cyklohexane The pure fractions were collected and evaporated to give a colourless gum. The gum was dissolved in THF (5 ml) and NH 4 0H aq (25 2 ml) was added and the mixture was stirred at 0,5 bar and 65°C over night. The solution was evaporated and the residue was purified on silica gel by elution with ethylacetae/methanol 8:1. Evaporation of the pure fractions gave a gum.

The gum was dissolved in CH 2 C1 2 (1 ral) and trimethylsilyl iodide (0.018 ml, 0.125 mmol) was added under nitrogen. The solution was stirred for 60 minutes at 20°C and evaporated.

The residue was dissolved in H 2 0 (1 ml) and washed with Et2O (2 x 2 ml). The aqeous phase was evaporated to give the title compound as a white solide.

1H-NMR Bruker 250 MHz D 2 0 20C: 1,4-2.4 ppm (CH 2 CIHN, CHCH20 (m) 6H 3.4-3.8 ppm CH 2 OH 4H 4.6-4,8 (CHN) 1H 5.8-5.9 ppm CH=CH 1H 7.7-7.8 CH CH 1H.

Example 10. Preparation of 1-[(3.4-trans-C-dihvdroxvmethvl)cvclooentvll-5methvlcytosine (compound The title compound was prepared from 1-[(3,4-trans-C-dibensyloxymethyl)-cyclopentyllthymine (compound 16) in the same manner as described for compound 14 in Example 9, :1 S S S 2 ExaMple 11 HO

HO

1-13 ,4-cis-C-dihydroxymnethyl(cycopentyllcytosifle NH 2

N-

Ais cm TLC-, EOAc/MveOH/H20 7:2:1; Silica iRf. 0.25

OR

*0 000e S

S

S@

3

S.

p S

.OS

SI S~"S

S

0O** 0* *5 0 S3 so S S i~L~) 1E.-NMR (D 2 57.65 1H, OH CH), 5.95 1H, OH CH), 4.75-4.65 (in, 1H, 3.70-3.60 (dd, 2H, CH 2 OH), 3,55-3.45 (cld, 2H, CH 2 OH), 2.35-1.35 (MI 6H, H-2'1 Example 12 1- [3 ,4-ci s- C-dihydroxymeQthyl (cycl open tyl Ithyrnine

HO-

A

HO Cl Ais

CH

3 TLC: EtOAcf~eOHfE 2 O 7:2:1; Silica RI': 0.35 I

S

A

IH-NMR (D120)-,87.55 lIH, C CH), 4,85-4.65 (mn, 1H1, 3.80-3.70, (dd, 2H, CH 2 3.65-3,55 (dd, 2H, CH 2 OH), 2.55-1.50 (in, 6E, H-4', 1.85 3H, CE 3 k -1 31 Preparation of intermediate products The starting materials for compounds 11, 17, 14 and 20 in Examples 7, 8, 9 and 10 respectively, were prepared in the following manner: The carbocyclic ring was built using the sodium cyamide dimerization of diethylmaleate described by Dolly et al. Cyclopentanone-3,4-dicarboxylique acid-dimethyl ester (compound 1) was dissolved in ethyleneglycol The solution was saturated with HC1.

Ketalization was let to proceed overnight at room temperature. The hydrochloric acid is then removed under 12 mmHg pressure, at room temperature. Distillation under reduced pressure 1 mmHg) allows 1dioxolano-3,4-dicarboxylic acid dimethyl ester to be isolated.

The above product is a 1:1 mixture ofcis and trans dicarboxylic acids dimethyl esters (compound which can be isomerized mainly (30 to the trans form under the following conditions. The ketal is dissolved in toluenemethanol 10 ml/g) and sodium methylate (0.3 eq) is added and the whole is warmed at 50oC for five hours. The reaction is then poured into water and extracted with ethylacetate to give compound 3.

13C-NMR (CDC13): 173.5 ester carbonyl; 114.9 0-1; 64.0 (t, JCH 147.74 H3) dioxolano; 51.6 JCH 147,74 H3) OCH3; 43.5 (d, JCH 173.3 H3) C-3 and C-4; 38.6 JcH 133.0 H3) C-2 and 1. L.J. Dolly, S. Esfandiari, C.A. Elliger, K.S. Marshall, J, Eng. Chem., 36, 1277 (1971), 2. B.G. Howard, R.V. Lindsey, J. Am. Chem, Soc., 82, 158 (1960) 1-Dioxolano-3,4-trans-dicarboxylic acid dimethyl ester (compound 3) is suspended in tetrahydrofuran (10 ml/g) and lithium aluminium hydride (0.32 g/g starting material) is added portionwise, with a fast stirring, S sufficiently quickly to keep a mild reflux. Stirring is pursued for I hour after the addition is complete. The reaction is then poured into ethylacetate and filtered over a Celite bed. After evaporation of the solvent, the crude material is used in the next step, 32 1-Dioxolano-3,4-trans-C-dihydroxymethyl (compound 4) (7.8 g, 44.6 mmol) is dissolved into THF-DMF mixture and added to a suspension of sodium hydride (2.81 g, 94 mmol) in the same solvent, under reflux.

Some 30 minutes later, neat benzyl bromide is added dropwise under reflux.

Reflux is pursued for 2 hours after addition is complete. After cooling, the reaction is poured into ethylacetate and thoroughly washed with water.

The crude dibenzylated material (compound 5) is then dissolved in dichloromethane, containing 15 isopropanol (5 ml/mmol) and toluene sulfonic acid monohydrate (1.05 eq) is added. Stirring is pursued for 4 hours, after which the reaction is poured into 0.01 N NaOH and extracted with dichloromethane. Purification by column chromatography, using hexaneethylacetate 1:1 as the eluent gives 1-oxo-3,4-trans-di-C-hydroxymethyl-3,4di-O-benzyl-cyclopentane (compound 6) (27.6 mmol, 9 g) in 62 overall yield.

1H-NMR (CDC13): S 7.30 10H) benzyl aromatic fractions; 4.5 4H) benzyl CH 2 3.5 4H) CH20; 2.5 4H) H-2 and H-5; 2.2 2H) H-3 and H-4.

13C-NMR (CDC13): 138,3, C-1; 127,9 and 126.8 benzyl aromatic carbons; 73.0, benzyl CH2; 72.2, -CH 2 41,7, C-3 and C-4; 38,9, C-2 and A solution of compound 6 (0.50 g, 1.54 mmol) in EtOH (30 ml) was 20 heated to reflux and KCO0 3 (0.300 g, 2.2 mmol) was added followed by

NH

2 0H x HC1 (0,26 g, 3.71 mmol) in small porti ns. The solution was cooled and diluted with H20 (100 ml) and extracted with CH2C12 (3 x 50 ml). The combined extracts were dried with Na2S04 and evaporated. The residue was dissolved in THF (50 ml) and LAH (0.23 g, 6.16 mmol) was carefully added.

The solution was refluxed for 5 hours cooled and quenched with H20 (1 ml).

The solution was diluted with Et 2 0 (200 ml) and filtered through celite, The solution was washed with H20 (3 x 20 ml), extracted with HC1 (aq) (3 x 20 ml), extracted with HC1 (aq) (3 x 20 ml 2M) and the aqeous phase was made alcaline with NaOH (aq) to pH 14.

30 The aqeous phase was extracted with Et 2 0 (3 x 20 ml), the combined extracts were dried with NaS04 and evaporated to give a pale yellow oil. The oil was purified by flash chromatography on silica gel (Merck Si 60) by elution with CH 2 Cl 2 /MeOH/NH3 95:3:2. The pure fractions were evaporated 33 to give ,4-trans-O-dibensyloxymethyl)cyclopentyl] amine (compound 8) as an oil.

1H-NMR Bruker 250 MHz, (0D01 3 1.3 2.5 ppm-OHCH2O, 2H, OH 2

CHNH

2 411, 3.3 3.7 CH 2 O, 4H, 4,4-4.7 Ph-OH 2 4H, OHNH 2 1H, NH 2 2H, 7.3 7.4 ppmn CrH,9-H 2 'ego 0 0 .o 0 Ogog 0 0 6 o go go C 06 o go go g 'egg 0 gogoog g 0 0 go 0000 0 0000 go o 04 00 0 0006 go go 0 0 0 0 0 BIOLOGICAL TESTS Test I Effects of compounds of the formula I on HIV in H19 .cells Table I Inhibition of human immunodeficiency virus in cell culture multiplication Concentration Inhibition Compc~z,,nds PM% 1-(3-fluoro-2,3 ,-dideoxy-a- (VSA 411) 0.1 1-(3-tluoro-2,3 ,-dideoxy-a- (VSA 409)* 2.5 1- 2,3-D ideoxy-3'C- (hydroxyme thylI)aL-D-erythro-pentafuranosyll cytosine (compound 21a) 1 5 l-[ 21 7 3 -Dideoxy-3'C-(hydroxymaethyl).

-D-erythro-pentafuranosyl] thy-mine (compound 22a)* 10 The tests were performed as described in the specification of the application: H9 cells, 105 cells per well on a 24-well plate, suspended in 2 ml RIPMI-medium containing 10 fetal calf serum, 100 ug/ml penicillin, ug/ml streptomnycin sulfate and 2 ug/ml polybrene are exposed to HIIV (HTLV- KIIB) and different concentrations of the test compounds. The plates are incubated at 370C in 5 CO 2 for 6 7 days. The contents in each well is then homogeneized with a pipette and transferred to a centrifuge tube. After centrifugation for 10 min at 1500 rpm the supernatant is removed and the cell pellet is analyzed by fixing in methanol on glass plates. Human HIV positive serum diluted 1:80 or 1:160 is added and incubated for 30 min at 37°C. The plate is then washed with phosphate-buffered saline (PBS) containing Ca2+ and Mg2+. Sheep antihuman conjugate (FITC) is added and after a new incubation the plate is again washed with PBS. Contrast staining is done with Evans blue and after drying the frequency of HIV antigen containing cell is determined in a microscope.

S

e

*S

S

S

S S S o• Vie _i ~I 1

I

36 Test II Cellullar toxicity Table 2. Cellular toxicity on H9 and F5000 cells.

(pM) Compound H9 F5000 1-(3-fluoro-2,3,-dideoxy-a-D-ribo- (VSA 411) 400 500 1-(3-fluoro-2,3,-dideoxy-c-D-ribo- (VSA 419) 250 1-[2',3'-Dideoxy-3'C-(hydroxymethyl)- 1 0 a-D-erythro-pentafuranosyl] thymine (compound 22a)* >100 H9 cells, 2 x 107 cells per plate, are incubated in RPMI-1640 medium containing 10 fetal calf serum, 70 mg/1 penicillin, 100 mg/1 streptomycin and 10 mM hepes, in absence or presence of test compounds. The number of cells per plate is determined after 48 hrs. Cells incubated in absence of test '15 compound then underwent two cell division cycles.

F5000 cells, which are human embryo cells, 1 x 105 cells per plate, are incubated in Eagle's minimal essential medium, supplemented with Earle's salts, non-essential amino acids, 10 fetal calf serum, 10 mM hepes, 70 mg/1 penicillin and 100 mg/ streptomycin, in absence or presence of test :20 compounds, The number of cells per plate is determined after 48 hrs, Cells incubated in absence of test compounds underwent one cell division cycle. The results are given as TC50, which is the concentration of a compound which gives 50 inhibition of cell multiplication.

It is seen in Table 2 that the test compounds exhibit TC50 values which vastly exceed the concentration needed for inhibition of HIV virus multiplication according to Table 1.

.i i i

Claims (10)

1. A catqoound of the formula:, R 3 R2 HO_ x and therapeutically acceptable salts thereof, wherein -the radicals A, 41 R 2 and R, 3 are as defined as follows, 0 KH 2 R R1 0 N N wherein is Hi; a~y1 contai ning 1-3 carbon atm-is, including or -c.cr-1 000 0 or CH 2 go 2 1H* and~ F H;V; Qll QCWt; or QU 2 Q-X wherein when R3 in formula I l0s F; l;1';CQ;C ocOI{t mnay have either, cis- coniuration or ttans.-configUtation relative to the hydroxyinethyl function at position with the proviso that when X is 0, then 0~ is not Hi. 38

2. A canpound according to claim 1 wherein A is KR wherein R1 is as defined in clai, 1,

3. A canpound according to claim 1 wherein A is wherein Rl is as defined in claim 4, A capond according t claim A

4. A compound accoding toQ claim F 8 .6 A ccnpound according to claim

7. A carvound accordig to anyi A cmpound according to claim hhdOXY~thl a Psition 41hav Bi 1, 2 or 3 wherein I is H or ai 3 or CiH 5 4 wherein RI is 4 wherein RI is H 3 ot' C2 ie of claims 1 to 6 wherein R3 is H; 7 wherein 3 's OC:$ 3 Or CHaC)U. 7 wherein is H 8 or 9 wherein R 3 at position 3' nd the trai:onf iguration, i' 'i 39

11. A process for the preparati-,, of a comnpound of the formula: R 3 R 2 JA 4i ca-ancrner or a therapeutically acceptable salt thereoi, wherein A, X, R, and R 3 are as defined in any one of claims 1 to 10 by: A, Condensing a glycoside as comprised in formula I above to the N-i position of a pyriinidine derivative corresponding to radical A in, foxmula Twhereafter the -anaper and the -ancner of cmpoizmd I thus formed are separated and any protectiLng groups removed; f:Ancmterization of a ,-anmr of the oua A R~ 2 Whe-roin A, and R 2 are s defined aIov( R4t sU ;C;C;0~3 o~ 2 0R; and. is H or a hydr ,v-rtectirn giroutoa utr of 'Aand -ancmers whereafter the -x-anrer and the a acie re -epar td and any protc-ting %zoups removed; A1 iS l: z 1 glycosylaticol of a n~ucleoside of the formula: eq 4 1 ii wherein X, R 2 R 4 arid R 5 are as def ined above, and B is a pydxnidin,: or purin base, for the formation, of a nucleoside containing the pyrimidine radical A as def ined above, wh(.-reafter the ct-ancmer is separated arnd any protecting groups removed; L: Substitution of the radical R 7 in a compound of the formula: 7LA V R wherein A, X and R are as defined above, and R7is a leaving group, With a radical R 3 whereafter any protecting groups are ren.wed; B: Conversion of the uracil moiety 0 inacTeido h oml I toactsiemit 0 *g whrein a c ompond of the formula tohascotos ine d ifoietydi coneredtoa heaputialy ccptbNHslt 41

12. A ccmpound or salt of freLmula I according to any one of claims 1 to when obtained by the process of claim 11.

13. A pharmaceutical composition comnprising a pharmaceutically-acceptable carrier or adjuvant and as active ingredient at least one ccmpound or therapeutically acceptable salt thereof of any one of claims 1 to 10 and 12.

14. A method for the therapeutic and/or prophylactic treatment of infections in mammals and man caused iy a retrovirus including HIV or hepatitis B virus, by administering to a host in need of such treatment an effective amount of at least one canpound or therapeutically-acceptable salt thereof of any one of claims I to 10 and 12, or of a pharmacutical comnposition of claim 13. A method according to claim 14 wherein the therapeutic treatment is for infections in man caused by HIV virus. C °S S g DATED this 13th day of June, 1991 MEDIVIR AB, its Patent Attorneys, A E. F, WELLINGTON CO., y~r .o S. Wellington) S S 5 S q .4 INTERNATIONAL SEARCH REPORT International Application No PCT,'SE88/00169 1, CLASSIFICATION OF SUBJECT MATTER (it several Claasific3tion symois apply, indicate aill) Accorjling to International Patent Classificatlon (IPC) or to both National Classification and IPC Lit C 07 H 19/06, 19/073; C 07 D 239/146, 405/014, 409/011; A 61 K 31/70 It, FIELDS SEARCHED K' ,nlmum Documentation Searched Classification System Classification Symbols IPC 2,3,14 C 07 H 19/02, 19/014, 19/06, 19/073; A 61 K 31/70 IPC 1 C 07 d 51/50, 51/52 us Cl 1536:23, 214; 424:180; 514:23, 42, 43, 49, Documentation Searched other than minimum Documentation to the E xtent that such Documents are included In the Fields SearchedI SE, NO, DK, FlI classes at, above III, DOCUMENTS CONSIDERED TO BE RELEVANT' Cateory I Citation of Document, 11 ith Inidication, where appropriate, of the relevant passages I? Relilivant to ciaim No. 13 x Journal of [Medicinal Chemistry, Vol 17, No 2~19714, 1-114, 17 T Kulikowski et al, "5-Alkylpyrirnicine Nucleosides, Preparation and Properlties of 5i-E~hyl-Z2-deoxy'cyti- dine and Related Nucleosides"', see P. 269-2173 X EP, A, 0 217 580 (THE WELLCOME FOUNDATION LIMITED) 1-114, 17 8 April 1987 EP, 0196185 DE, 3608606 JP, 62103100 X DE, A, 3 002 197 (ROBUCEN GMBH PHARMAZEUTISCUE 1-114, 17 FABRIK ESSLINGEN A,N. 23 July 1981 xDE, A, 3 0145 375 (ROBUGEN GMBH PHARMAZEUTISCHE 1-114, 17 FABRIK ESSLINGEN AIN.) 1 July 1982 X D8, A, 3 229 169 (ROBUGEN GMBH PHARMAZEUTISCHE 1-114, 17, F'ABRILK) 9 Febriuary 1984 *Special categories of cited documents: It 'IT" later document published after the International tiling date "A"l document defining the general state of the art which to not or Priority dae end not in conflict with the application but cited to understand the principie or theory underlying the c,otred ,o *e of particular feisee4nce Invention "Ell eartii,: document but publiehed on or atter the international document of perlcular relevance; the claimed invention fiiing dale cannot be considered novel or cannot be Considered to I'Ll document which may throw doubts on priority ciatmlat or Invotve an inventive stop which Is cited to establish the publication date of another "Yt" document of particular refevance; the claimed Invention citation or other special reason (as specifledl cannot be Coneidered to involve an Inventive stop when the 110" document reterring to an opal disclosure, use, exhibition or document Ie combined with one or enore other such docu* other means maonta, such combination being obvious to a person skilled document published prior to the International filing date but In the art. later then the priority date claimed document member of the ame patent faily IV, CERTIFICATION Datcl of the Actual Completion of the International Search I Dale of Mailing of this Internatltinal Search Report

1988-07-11 '1988 -07- 2 2 International Searching Authority ISi gnature~1," Ithitsed Oficer SWedish Patent Office GnlaCaso Form PCT/lSAi2lO0 (eecond ahoot) (January 19a5i 7International Anoi~cal.,n No. PCT/SE88/00169 FURTHER INFORMATION CNIUDFROM THE SECOND SHEET VT OBSERVATIONS WHERE CERTAIoN' CLAIMSWEVE FOUND :irtUA This nteralioniserch epor hs hot been establiihed in resot~~i of cqrteln ci~ar.it undet Ar-tic- 17(2) for the following reasons: IE.Claim nummers 45. i cu se they relate to aubieci matte~r not recuirea e 0- to~ n a~u by lnis Autnority, namely: Method for treatment of the human or, animal body by therapy, 2Q~ claim numbern berauce they reiate to Paris of th," nternatuinim awrilictuo10 irtl co -rot comoiy with the prescribed require- menua to such an extent trial no fneaningiul internaiont. seattri can IV c7Wru.!.' 0 r-ctz *Claims 1-Th and 17 has been searched incompletely. According to PCT, Art, 15(3) the search has been limited to wha±t has been shown in the working examples, namely compounds of the formula 1 wherein X is 0, 3 ,claim ntumes oeCaute they are oepamnft claims ano ara not cfyaW in z700.janCe v.iff ti~e socon4 a" trurd wetences of FPCT RuiW 6,4(a) OU11SERVATIONS WHeIE UNITY OF VNNNTION IS LACKtING Thin International Searching Authority found multiple invention% In thok 'nternatOns apuplication its follows:z 1.7 As all required additional search tees were timely paid by ttia applicant, this International aearcrn report covers all searchable claims of the International applicafidri, 42.[D As only some ot In* required additional soarc, Ito were timeiy paid by the apDlican', Iris intpmnationsl search report covers only those claims of the lntonafionai application for which fees were paid, Specificaily ciaimr. 3.E] No (squired additional search toes were timniy paid by the Cotif riiianuoucrn!!% trit invornational search report Is restricted to the Invention lirbt mefltiohcid It. the ctaime; It is coverso by claim riumooe 4,)As all. sarctiebieclaims could ba searched Without eflort justifying at, aoojtitna tte t-r iiernatioflal Seanrchinig Authority did not Invite payment of any additional feei Romarix on Pr'0 1 ,st EThe additional search Ite were accompanied by apot\ ont's aroto t No protest accompanied the Payment of additional sear4 te01 LForm PCTIISAI (supplemenital &heet (January tOaS International Application 1, PCT/SE88/OO169 III, DOCUMENTS CONSIDERED TO BE RELEVANT (CONTINUED FROM THE SECOND SHEET) Category Citation of Document, witn indication, wtwvr aPooPfite, 01 "i re4.lfl) pasagesa~ Relevant to Cla-m No X US, A, 3 116 282 (THE UPJOHN COMPANY) 31 December 1963 X Chemical Abstracts, Vol 102 (1985) abstract No 7013q, Chem. Pharm. Bull. 1984, 32(4), 14111-50 (Eng) X Chemical Abstracts, Vol 92 (1980), abstract No 2O89Q3d, Vopr. Virusol. 1979, 603-6 (russ) X Chemical society, London. Journal. Perkin trans., 1978, P J Barr et al, "The Synthesis of Nucleosides derived from 5-Ethynyluracil and see p. 1263-1267 X DE, A, 2 930 9011 (GAURI, KAILASH KUMNAR) 19 February 1981 X DE, A, 2 918 260 (ROBUGEN CM5H FABRIh ESSL.INORN AAN) 27 November 1980 X EP, A, 0 206 497 (THE WELLCOME December 1986 i, 61280500 PHARMAZ EUTISCH-E FOUNDATION LIMITED) 1-14, 17 1-14, 171 1-14, 1 1-14, 17 1-14, 17 11,17 1-14, 17 1-14, 17 1-111, 17 1-14, 1*7 X EP, A, 0 196 185 (THE 1 October 1986 DE, 3608606 EP, 01994I51 JP, 61257925 US, 4724232 EP, 0217580 GB, 21181128 JP, 62103100 WELLCOME FOUNDATION LIMITED) X EP, A, 0 199 451 (THE WELLCOME FOUNDATION LIMITED) 29 October 1986 JP, 61257926 EP, 0196185 DE, 3608606 JP, 61257925 US, 4724232 X US, A, 3 817 982 (VERHEYDEN ET AL) 18 June 1974 X US, A, 4 247 544 (BERGSTROM ET AL) 27 January 1981 X US, A, 4 267 171 (BEROSTROM ET AL) 12 M"ay 1981 Form PCT ISA 210 (extra &hoest) (January 1"51, H I. international Apolication %:PCT/SE88/00169 III, DOCUMENTS CONSIDERED TO BE RELEVANT (CONTINUED FROM THE SECOND SHEET) I Category Citation of Document, witri indication, wtiei' aporopfIate. of "e realavani oasages Relevant to Clim No X J. Carbohydrates. Nucleosides, Nucleotides, Vol. 5, No 3, 1978, E DeClerq et al, "Nucleoside analogs with selective antiviral activity' see p 187-224l x FR, A, 2 0140 177 (ROBUGEN GMBH) 22 January 1971 X DD, A, 75 084 (DR GERHARD ETZOLD ET AL) August 1970 1-1~4 1-114, 17 1-114, 17 I ~~orm PCT ISAZt Wtta a heet) (JatuaIV 1945)