CA2481078C

CA2481078C - Method and compositions for the synthesis of bch-189 and related compounds

Info

Publication number: CA2481078C
Application number: CA002481078A
Authority: CA
Inventors: Dennis Liotta; Woo-Baeg Choi
Original assignee: Emory University
Current assignee: Emory University
Priority date: 1990-02-01
Filing date: 1991-01-31
Publication date: 2009-12-15
Anticipated expiration: 2011-01-31
Also published as: JP2001019690A; AU4031995A; AU4474599A; EP0513200A4; NO970385D0; CA2678778A1; FI20060622A; AU2002300661B2; DE69133556D1; RU2125558C1; EP0513200B2; FI121069B; MC2233A1; RO108564B1; EP0872237A1; JP3844978B2; NO326249B1; NO970385L; HU211300A9; DE122004000015I2

Abstract

The present invention relates to a method of preparing BCH-189 and various analogs of BCH-189 from inexpensive precursors with the option of introducing functionality as needed. This synthetic route allows the stereoselective preparation of the biologically active isomer of these compounds, .beta.-BCH-l89 and related compounds.
Furthermore, the stereochemistry at the nucleoside 4' position can be controlled to produce enantiomerically-enriched .beta.-BCH-189 and its analogs.

Description

METHOD AND COMPOSITI~NS FOR THE SYNTHESIS OF BCH-189 .. _ ~ t' ~D RELATED COMPOUNDS
.:
The present invention relates to methods and ~ w compositions for preparing antiviral nucleoside analogs, -particularly BCH-189 (2°,3'-dideoxy-3°-thia-cytidine).
More particularly, the invention relates to the selective ~ ' synthesis of the B-isomer of BCH-189 and related compounds as well as the selective synthesis of enantiomerically- .
enriched BCH-189 and related compounds. - ', ~~G~UND ART
In 1981, documentation began on the disease that became known as Acquired. Immune Deficiency Syndrome (AIDS), as well as its forerunner AIDS Related Complex ~-(ARC). Tn 1983, the cause of the disease AIDS was ~.
established as a virus_ named the Human Immunodeficiency Virus type 1 (HIY-1). usually, a person infested With the virus will eventually develop AIDSt in all known cases of AIDS the final outcome has always been death.
The disease AIDS is the end result of an HIV-1 virus following its own complex life cycle. The virion -life cycle begins with the virion attaching itself to the host human T-4 lymphocyte immune cell through the bonding of a glycoprotein on the_surface of~the virion°s protective coat with the CD4 glycoprotein on the lymphocyte cell. Once attached, the-virion sheds its glycoprotein coat, penetrates into the membrane of the host cell, and uncoats its RNA. The virion enzyme, reverse transcziptase, directs the process of transcribing the RNA into s3,ngle stranded DNA. The viral RNA is degraded and a second DNA strand is created. The now ., double-stranded'DNA is integrated into the human cells -genes and those genes,are used for call reproduction. --~., 2.
At this point, the human cell carries out its reproductive process by using its own RNA polymerise to -transcribe the integrated DNA into viral RNA. The viral ~:' ~tNA is translated into glycoproteins, structural proteins, and viral enzymes, which assemble with the viral RNA
~, intact. When the host cell finishes the reproductive step, a new virion cell, not a T-~ lymphocyte, buds forth. ~ .;
The number of 8IV-1 virus cells thus grows while the ~w,~. .
number of T-4 lymphocytes decline.
The typical human immune system response, .
killing the invading virion, is taxed because a large portion of the virion's life cycle is spent in a latent state within the immune cell. Zn addition, viral reverse . .
transeriptase, the enzyme used in making a new virion cell, is not very specific, and causes transcription mistakes that result in continually changed giycoproteins .
on the surface of the viral protective coat. This lack of specificity decreases the immune systems effectiveness . .w because antibodies specifically produced against one glycoprotein may be useless against another, hence reducing the number of antibodies available to fight the . ' virus. The virus continues to grow while the immune response system continues to weaken. 8vantualiy, the BTv largely holds free reign over the body~s immune system, allowing opportunistic infections to set in and ensuring that, without the administration of antiviral agents - , andjor immunomodulators, death will result.
There are three critical points in the virus's lice cycle which have been identified as targets for antiviral drugs: (1, the initial attachment of the virion to the T-4 lymphocyte, or aacrophage, site, (Z, the transcription of viral Rte to viral Dot, and ( 3 ) the 35. assemblage o! the new virion aall during reproduction.

Inhibition of the virus at the second critical point, the viral RNA to viral DNA transcription process, has provided the bulk of the therapies used in treating AIDS. This transcription must occur for the virion to ,;
reproduce because the virion's genes are encoded in RNA; w the host cell reads only DNA. By introducing drugs that .' block the reverse transcriptase from completing the formation of viral DNA, HIV-1 replication can be stopped.
l0 Nucleoside analogs, such as 3'-a2ido-3'-deoxythymidine (AZT), 2',3'-dideoxycytidine (DDC), 2',3'-dideoxythymidinene (D4T), 2',3'-dideoxyinosine (DDI), and . various fluoro-derivatives of these nucleosides are relatively effective in halting gZV replication at the .~ ~.
reverse transcriptase stage. Another promising reverse ~ .-~ -transcriptase inhibitor is 2',3'-dideoxy-3'-thia-cytidine . '~ ,~
(BCH-189), which contains an~oxathiolane ring substituting ..~ ~.-_ for the sugar moiety in the nucleoside. ~
AZT is a succsssful.anti-AIV drug because it sabotages the formation of viral DNA inside the host T-4 lymphocyte cell.' When AZT enters the cell, cellular kinases activate AZT by pbosphorylation to AZT
triphosphate. AZT triphosphate then competes with natural thymidine nucleosides for the receptor site of HIV reverse transcriptase enzyme. The natural nucleoside possesses two reactive ends, the first for attachment to the ' previous nucleoside and the second for linking to the next :.
nucleoside. The AZT molecule has only the first reactive ends once inside the HIV enzyme site, the AZT aide group terminates viral DNA formation because the azide cannot make the 3',5°-phosphodiester with the ribose moiety of . .
the following nucleoside.
AZT's clinical benstits include.increasad ' ~.
longevity, reduced frequency and severity of opportunistic infections, and increased peripheral CD4 lymphocyte count. ;;~...
._..;
Y r. a .' .y:: T ' : . ' ~ F . ' ~,~lt ~., ~' , , . . . t~ . ', . . .. , ... .
. . . . , . ..
..., ~.' ' : . - ;.~ ~ Z. ~ . :. . ~,~ ,~,.. . ~j~:w~~',,.. . ~ \~' ' ' ~.~
~.. ..:.' ' ,:.- . ~~ : .

Immunosorbent assays for viral p24, an antigen used to track HIV-1 activity, show a significant decrease with use .
of AZT. However, AZT's benefits must be weighed against :y y' the severe adverse reactions of bone marrow suppression, nausea, myalgia, insomnia, severe headaches, anemia, peripheral neuropathy, and seizures. Furthermore, these .
adverse side effects occur immediately after treatment . --begins whereas a minimum of six weeks of therapy is necessary to realize AZT's benefits.
Both DDC and D4T are potent inhibitors of HIV
replication with activities comparable (D4T~ or superior . . y (DDC) to AZT. however, both DDC and D4T are converted to their 5' triphosphates less efficiently than their natural ' analogs and are resistant to deaminases and ' phosphorylases. Clinically, both compounds are toxic.
Currently, DDI is used in conjunction with AZT to treat ~ w AIDS.. However, DDI's side effects include sporadic pancreatic and peripheral neuropathy. Initial tests on . ., 3'-fluoro-2'-3'-dideoxythyzaidine show that its anti-viral activity is comparable ~to that of AZT. w Recent tests on BCH-189 have shown that it possesses anti-HIV activity similar to AZT and DDC, but without the cell toxicity which causes the debilitating .side effects of AZT and DDC. A sufficient quantity of ~ ~ . .' , BCH-189 is needed to allow clinical testing and treatment using the drug.
~ The commonly-used chemical approaches for ~ .
synthesizing nucleosides or nucleoside analogs can be W'~'--classified into two broad categories: (1) those which - .
modify intact nuceosides by altering the carbohydrate, the . , bass, or both and (2) those which modify carbohydrates and ..
incorporate the base, or its synthetic precursor, at a suitable stage in the synthesis, Bsvauss HCH-~1s9 substitutes a sulfur atom for a c~rrbon atom in the carbohydrate ring, the second approach is more feasible.
The most important factor in this latter strategy involves delivering the base from the B-face of the carbohydrate ring in the glycosylation reaction because only the 8- ' 5 isomers exhibit useful biological activity.
It is well known in the art that the stereoselective introduction of bases to the anomeric centers of carbohydrates can be controlled by capitalizing ..- ..
to on the neighboring group participation of a 2-substituent on the carbohydrate ring (. em. Ber. 114:1234 (1981)).
However, BCH-189 and its analogs do not possess a 2-substitutent and, therefore, cannot utilize this procedure unless additional steps to introduce a functional group that is both directing and disposable are incorporated into the synthesis. These added steps would lower the overall efficiency of the synthesis. .
It is also well known in the art that 2~ "considerable amounts of the undesired a-nucleosides are always formed during the synthesis of 2'-deoxyribosides"
CChem. . 114x1234, 1244 (1981)). Furthermore, this reference teaches that the use of simple Friedel-Crafts '.
catalysts like SnCI~ in nucleoside syntheses produces undesirable emulsions upon the workup of the reaction mixture, generates complex mixtures of the a and b-isomers, and leads to stable o-complexes between the SnCl4 and the more basic silyated heterocycles such as silyated cytosine. These complexes lead to longer, reaction times, lower yields; and production of the undesired unnatural N-3-nucleosides. Thus, the prior art teaches the use of trimethysilyl triflate or tximethylsilyl pdrchlorate as a catalyst during the coupling of pyrimidine bases with a carbohydrate ring to achieve high yields of the biologically active 6-isomers. However, the u'ss of these catalysts to synthesise BCH-189 or BCH-189 analogs doss a.
:3: ...

not produce the b-isomer preferentially: these reactions , r~sult in approximately a 80:50 ratio of the isomers.
Thus, there exists a need for an efficient synthetic route to BCH-189 and its analogs. There also ~.
exists a need for a stereoselective synthetic route to the .
biologically active isomer of these compounds, S-BCH-189 and related 8-analogs. Furthermore, there exists a need for a stereoselective synthetic route to enantiomerically-enriched B-8CH-189 because the other enantiomer is inactive and, therefore, represents a 50% impurity.
n=scro~q~~a og =~rrr=o~
. . ., The present invention relates to the discovery of a surprisingly efficient synthetic route to BCH-189 and ' various analogs of BCH-189 from inexpensive precursors with the option of introducing functionality as needed.
This synthetic route allows the stereoselective preparation of the biologically active isomer of these ., compounds, 8-BCH-189 and related compounds. Furthermore, the steochemistry at the nucleoside 4~ position can be ~,_ controlled to produce enantiomerically-enriched t~-BCH-189 and its analogs. .
2 5 ~ , ys' .
The term "BCH-189 analogs" is meant to refer to_ nucleosides that are formed from pyrimidine bases substituted at the 5 position that are coupled to substituted 1,3~oxathiolanes.
The method of the present invention includes ozonizing an allyl ether or ester having the formula . ' CHz~iCH-CH=-OR, in which R is a protesting group, such as an ~ .
' alkyl, silyl,~or aryl group-, to form a glycoaldehyda ...' ._ having tb~ foraula o~iC~t-OR: adding thioglycolic acid to ~~
the glyooaldshyde to form a lactone of the formula 2-(R- ~w.'.
oxyj-methyl-5-oxo-1,3~oxathiolanes converting the lactone to its corresponding carboxylate at the 5 position of the oxathiolane ring; coupling the acetate with a silyated pyrimidine base in the presence of SnCl~ to form the a-isomer of a 5'-(R-oxy)-2',3'-dideoxy-3'-this- nucleoside analog; and replacing the R protecting group with a hydrogen to form BCH-189 or an analog of BCH-189.
The invention can be used to produce BCH-189 or BCH-189 analogs that are enantiomerically-enriched at the l0 4' position by selecting an appropriate R protecting group to allow stereoselective selection by an enzyme. For instance, the It protecting group can be chosen such that the substituent at the 2 position of the~oxathiolane lactone is butyryloxy to permit stereoselective enzymatic ~15 hydrolysis by pig liver esterase. The resulting optically active hydrolyzed lactone can then be converted to its corresponding diacetate and coupled with a silyated pyrimidine base as above. . ,...
20 Accordingly, one of the objectives of this invention is to provide an efficient method for preparing the 8-isomer of BCH-189 and analogs of BCH-189 in high ~-yields. Furthermore, it is an objective of this invention to provide a synthetic method to produce only one optical 25 isomer, rather than a racemic mixture, of BCFi-I89 and w analogs of BCH-189. A further object of this invention is w to provide a synthetic route to produce B-BCH-189 that is enantiomerically-enriched.
3o Additionally, an objective' of this invention is to provide intermediates from which BCH-189 or BCH-189 . analogs can be synthesized of the formula 2-(R-oxymethyl)-5-acyloxy-1,3-oxat3iiolane, wherein R is a protecting group, such as alkyl, silyl, or acyl, and a method of 35 preparing these compounds. Furthermore, it is an object of this invention to provide enantiomsrically-enriched 2- ..~' _~~ .
ac~toxyiasthyl-5-acstoxy-1,3-oxathiolane and 2- , .

butoxymethyl-5-oxo-1,3-oxathiolane and methods of - preparing these compounds. , _.
Another objective of this invention is to provide intermediates from which BC~i-189 or BCH-189 analogs can be synthesized of the formula:
NHZ
N, Y -to RO ~
S
wherein R is a protecting group, such as alkyl, sibyl, or acyl, and Y can be-hydrogen, methyl, halo, alkyl, alkenyl, alkynyl, hydroxalkyl, carboxalkyl, thioalkyl, selenoalkyl, phenyl, cycloalkyl, cycloalkenyl, thioaryl, and selenoaryl, and methods of preparing these coumpounds.
- . . . _ :~.,:
Furthermore, this invention provides .
intermediates from which BCH-189 or BC~i-189 analogs can be synthesized ot~the formulas . H p ~ Y
RO ~ N' 'SJ . . .
a wherein R is a protecting group, such as alkyl, silyl, or acyl, and Y can bs hydrogen, methyl, halo, alkyl, alkenyl, alkyryl, hydroxalkyi, carboxaikyi, thioalkyl, selenoaikyl, phenyl, cycloalkyl, cycloalkanyl, thioaryl, and selsnoaryl, and methods of preparing these caumpounds. ~ ~~y~.: -:., , , .. . ; '. . :;_ : ~~ -_ r' : _ : _. - ~ . . ~ .:.,, :.:;: . .~ '- .;
,. :.. . :.

BRIEF ~1,~'~3c'AYpTIOZt of ~,n Wi~as Figure 1 illustrates one embodiment of a synthesis of HCti-189 and HCH-189 analogs according to the present inventian;
Figure 2 illustrates one embodiment of the synthesis of HCH-189 according to the present invention:
Figure 3 illustrates one embodiment of the synthesis of 5-methyloytidine and thymidine derivatives of BeFI-189 according to the present invention; and Figure 4 illustrates one embodiment of the synthesis of enantiamerically-enriched BCIi-189 according to the present in~rention. ' ..
H$sT H~s~ o~R~~~ ~ =~irrroN . -BCH-189 is a comgound of the formula:

N. i ...
HO O~N

S ., The process of the present invention for preparing HCii-189 and BCH-189 analogs is set forth in Fig.
1. An allyl ether or ester. is ozonized to give an . 30 aldehyde ~,, which reacts with thioglycolic acid to give a . v lactone ~,. The lactone ~, is treated with a reducing agent, followed by a carboxylic anhydride, to produce the . : _--carboxylate g. This aarboxylate is coupled with a silyated pyrimidine bass irr tha~presence of a Lewis acid .
.35 that,can~catalyae stsreospecific coupling, such as SnCh~, to yield the 8-isomer of the substituted nucleoside $ in essentially a 100:0 ratio of Bra isomers. The substituted ~~w 3:
.... .... .. .:: . . . . .. . . ~ .. . ~ . ~ .,. . . .: . ., .
;~.:,r:. ~. w.:. ~, :v: ~;... : .'' .:: : :: :;~ .; . ., ,~ y: :.. . : . , .
:.:. : ::..: . ~.. .:. .. w . . ;. ,, :.: ; ~ ~' : ~ .. ~ v . . ~ ~ ,.:
.: .. ' :. .. _ ..,: ~.:.. ; \; ,. ' ,. :' -: , ;'.. . ' ~ '".: ... . .:, nucleoside ~, is deprotected to produce BCH°-189 or BCH-189 analog ~. ~' This procedure can be tailored to produce BCH-5 189 or BCH-189 analogs that are enantiomerically-enriched at the 4' position by selecting an appropriate R
protecting group to allow stereoselective enzymatic hydrolysis of ~, by an enzyme such as pig liver esterase, porcine pancreatic lipase, or subtilisin or other enzymes 10 that hydrolyze ~, in a stereoselective fashion. The resulting optically.active ~, can be converted to enantiomerically-enriched carboxylate g and coupled with a silyated pyrimidine base as above to produce .; ;. .
enantiomerically-enriched 8CH-189 or BCH-189 analogs.
The protecting group R in ~, can be selected to provide protection for the corresponding alcohol until the final step in the synthesis is carried out (deprotection of ,~ to form ø). Additianally, the protecting group can ~.
2o be selected, if desired, t~ provide an additional -recognition site for an enzyme to be used later in an enantio-selective hydrolysis reaction. Any group that .
functions in this manner may be used. For instance, alkyl, silyl, and acyl protecting groups or groups that possess substantially the same properties as these groups can be used.
An alkyl protecting group, as used herein, means triphenylmethyl or an alkyl group that possesses 'substantially the same protecting properties as .
triphenylmethyl. A silyl prbtecting group, as used herein, means a trialkylsilyl group having the formula:
Ri . .... .
3 5 ~-~1-RZ . . .
R3 ., wherein R~, R=, and R3 may be lower-alkyl, e.g., methyl, ethyl, butyl, and alkyl possessing 5 carbon atoms or lease or phenyl. Furthermore, R~ may be identical to RZr R~, RZ, and R3 may all be identical. Examples of silyl protecting groups include, but are not limited to, trimethylsilyl and . ' t-butyldiphenylsilyl.
An aryl group, as used herein to describe an . ~ ' aryl protecting group (as in ~) or to describe a l0 carboxylate (as in 4), is a group having the formulas ..
O --R

wherein R' is a lower alkyl, e.g., methyl, ethyl, butyl, and alkyl possessing 5 carbon atoms or less substituted lower alkyl wherein the alkyl bears one, two, or.more . . .

20 simple substituents, including, but not limited to, amino, . .-carboxyl, hydroxy, phenyl, lower-alkoxy, s.g., methoxy and . .

ethoxy: phenyl: substituted phenyl where3.n the phenyl bars one, two, or more simple substituents, including, but not limited to, lower alkyl, halo, e.g., chloro and :' .

25 bromo, sulfato, sulfonylaxy, carboxyl, carbo-lower-alkoxy, e.g., carbomstboxy and carbethoxy, amino, mono- and di-lower alkylamino, e.g., methylamino, amido, hydroxy, lower alkoxy, a.g., methoxy and ethoxy, lower-alkanoyloxy, e.g., acetoxy. ,..

30 '.

A silysted pyrimidine base, as used herein, ' means a compound having th~ formula:

. ;:: . .

X w 35 . N Y .

ZO~N'~ ' .
. ;: . .
. .

;.:._.: ;..;:;-.,,.;.~::~ ..: :.::: . ~. ': . ';;.....:,:;::. . . .:...,:;..
,,,..:;,,.:.::.::::.. . . :~:':~;:,;.....;:.. . ,, ,. ... .:' :..
. :: .. .--.. . " :... ::. '.~ 'i irf: : ' ' :' . : . ., ..~. . .
. . .. .

.. .: . , . - . ... . y :: , " ' ' . . ~
.; .~ ~',.~:. :.: ;.. ...; ' ." w ', . ~ : ':: ..:1: .:: '..
: y . w..'.. ~ . ~ ...~.: . : .. '..- '' ;_, ' .. . ~ . ..
.';:,;
,' ...' ': ~

wherein X is either a trialkylsilyloxy or a trialkyisilylamino group, Z is a trialkylsilyl group, and Y is further described below. A trialkylsilyl group, as used herein, means a group having the formula: ' ~-st-R2 .

' wherein R~, RZ, and R3 may be lower-alkyl, e.g., methyl, ethyl, butyl, and alkyl possessing 5 carbon atoms or less, or phenyl. Furthermore, R~ may be identical to RZ: R~, Ri, and R3 may. all be identical. Examples o! trialkylsilyl, groups include, but are not limited to, trimethylsilyl and t-butyldiphexiylsilyl. ' ~ .
The silyated pyrimidine base may be substituted , with various Y substituents, including, but not limited 2o to, hydrogen, methyl, halo, alkyl, alkenyl, alkynyl, hydroxyalkyl, carboxyalkyl, thioalkyl, selenoalkyl, phenyl, cycloalkyl, cycloalkenyl, thioaryl, and selanoaryl, at position 5 0! the silyatsd pyrimidine base (Y substituent in Fig. 1) to modify the properties, such as transport properties or the rate o! metabolism, of the BCFi~189 analog.
illustrative examples o! the synthesis of EcH-189 or BCH-i89 analogs according to the present invention are given in Figs. Z, 3, and 4 and the following descriptions. , Figure 2 shows the synthmsis'o! BCH-189 starting with allyl alcohol Z. A Na8 oil suspension (4.5 g, 60~, 3 S 110 amol ) was washod with TEF' tarics ( 100 nl ~ x 2 ) assd the .
rssuitiiig solid suspended in T'HF (300 gal) . The suspension was cooisd to o~C, allyl aloohol Z (6.8 ml, l00 amol) was ~~.. ...~:~.' ~ ~:~ S.~':'',.' ~'.:. ~.~'S~. '.. ':.' .. .. ' ,~ ~ '~ ' .:
..'~. :-. , ." ,:'::'_ :. ;_.., ..' :', i.: ~..', .;~,'..: . . . , _. ..
~t ZS ' . _ . .. _ , _ . .~' :Sy: ~ . '.S'.'..' ~'~ ~ , .
t,~.~ f .. _ i ~., ; ~,y~ ~,~. .:',. .,..: .'..'.. .: '; . . ' '... .. . :~~. :.; ~~ ~
:.:,~'..'.' ' .~..~.'.',,'r!:::.:Y,~. ~s~~si. :~ :'.::.'.;~:...:', .. :. '.
... C' ' . . , , . ' . S ~ . . '. . .. . ~ , _. , ' ' . ~ . 1 , . . . :: ': . . ' 1.3 added dropwise, and the mixture was stirred for 30 minutes at 0'C. t-Butyl-diphenylsilyl chloride (25.8 ml, 100.8 mmol) was added dropwise at 0'C and the reaction mixture was stirred for 1 hour at o'C. The solution was quenched ..
with water (100 ml), and extracted with diethyl ether (zoo ml x z). The combined extracts were washed with water, dried over MgS04, filtered, concentrated, and the residue w distilled under vacuum (90-100'C at 0.5-0.6 mm Hg) to give a colorless liquid $. (z8 g. , 94 mmol, 94~) . (~H Nl~t: 7.70- . . ' '7.35 (lOH, m, aromatic-H) t 5.93 (iFi,, m, HZ) : 5.37 (1H, dt, H~) J$1.4 and 14.4 Hz; 5.0? (iH, dt, Hy) Jsl.4 and 8.7 tiz 4.21 (2H, m, R;): 1.07 (9H, s, t-Bu)) , .' The silyl allyl ether $ (15.5 g, 52.3 mmol) was ~ . w dissolved in CHzClz (400 m1), and ozonized at -78°C.- Upon completion of ozonolysis, DMS (15 ml; z04 mmol, 3.9 eq) was added at -78'C and the mixture was warned to room temperature and stirred overnight. The solution was washed with water (100 ml x z), dried over Mg80~, filtered, ' concentrated,' and distilled under vacuum (loo-110'C at 0.5~0.6 mm Hg) to give a colorless liquid ~ (15.o g, 50.3 mmol, 96~). (tH NMR: 9.74 (1H, s, B-CO)f 7.70-7.35 (lOH, '..
m, aromatic-H)t 4.21 (zH, s, -CHi): 1.22 (9H, s, t-Bu)) w Silayted glycoaldehyde ~ (15.0 g, 50.3 mmol) Was w dissolved in toluene (z00 ml) and thioglycolic acid.(3.50 . ml, 50.3 mmol) was added all at once. The solution was refluxed for z hours while the resulting water was removed : .
with a Dean-Stark trap. The solution was cooled to room temperature and washed with saturated NaHC03 solution and the aqueous washing: were eutsactod with diethyl ether (200 ml x ~). Ths combined extracts wars, washed with . water (1o0 ml x 2), dried over Mg80;, filtarsd, and eonaeutratad to give a colorless oil ~Q (16.5 g, 44.3 _ .
3S msol, 88~~, which gradually solidified under vacuum. .
Rserystailisation from hexane atfordad a vhit~ solid a,,Q
(15.8 g, 84t) . (~H i~t: 7.2~7.38 (10H, $, aromatic-H) s . . .
>. . , ,_ . ~t.t.~'~f.
.y . .. :. :: ~..
y; . . :
~:..
' ~
~

: , . .
' . . '.' , ~ ~. ,.
~ '.., . ~IA:
~. .._ ..
. . . ! ' ..

. . .' ::' ~ . . ' ~ . . ~ ! .. , , ' ii. '. s .
. . . ' . ~ : .
.
, ., r .;'.:. , i.':-.' . .
. ,. .. . . ~ . ~ . .
,i : ':~ ;y , i , :,. : .'.,. _ .' . ~. . .. ' .. . .. .. '. ". .~, ~' ~ ~':.
:' :~., ", ..' _ ' , . . : .. y. , '..
,: . ', . 53 ( 1H, t, ~I2) J=2 . ~ HZ i 3 . 93 ( 1H, dd, -G$Zd) aT~9. 3 IiZ i , 3 . 81 ( 1H, d, 1H~,) J=13 . 8 Hz i 3.79 ( 1H, dd, -G~IZO) ; 3 . 58 (IH, d, 1H4)i 1.02 (9H, S, t-Bu)) 5 2-(t-Butyl-diphenylsilyloxy)-methyl-5-oxo-1,2-oxathiolane ~ (5.0 g, 13.42 mmol) was dissolved in .
toluene (150 ml) and the solution was cooled to -78'C. ,.
Dfbal-li solution (14 ml, 1.o M in hexanes, 3.4 mmol) was .
added dropwise, while the inside temperature was kept below -70'C all the time. After the completion of the addition, the mixture was stirred far 30 minutes at -~8~G. ' Acetic anhydride (5 ml, 53 mmol) was added and the mixture was warmed to room temperature and stirred overnight. _ Water (5 ml) was added to the mixture and the resulting mixture was stirred for 1 hour at room temperature. The mixture was diluted with diethyl ether (300 ml), MgSO' (40 g) was added, and the mixture was stirred vigorously for 1 hour at room temperature. The mixture was filtered, ' concentrated, and the residua flash chromatographed with 208 EtOAc in hexanes to give a colorless liquid ,yl, (3.60 .
g, 8.64 mmol, 648), which was a 6:i mixture of anomers. y ( ~~i NMR of the ma j or isomer: 'i .'7 0-' . 35 ( lOH, m, aromatic- . .
H) ; 6.63 (18, d, 85) Js4.4 ~Hz: 5.47 (1H, t, ~I2) : 4.20-3.60 ~ , . .
(2H, m, -CHiO): 3.27 (1H, dd, 1~) Jm4.4 and 11.4 Hzi 3.09 ..
(IH, d, IH4) J~11.4 Hz i 2.02 (3H, s, C83CO) ; 1.05 (9H, s, t-8u) i ~H Nl~t of the minor isomers 7.70-7.35 (10H, m~, aromatic-H) i 6 .55 ( 1H, d, H5) Ja3 ~ 9 ~3Z i 5.45 ( lli, t, HZ) i 4.20-3.60 (28, m, -CFiZO) : 3.25 (IX, dd, 18~) J=3.9 and 11. 4 HZ: 3.11 (1H, d, lH,~j J~11.4 HZ: 2.04 (3~i, s, CH;CO) i 1.04 (98, s, t-Bu)) Z-(t-Butyl-diphenyleilyloxy)-methyl-5-aaetoxy- ' 1, 3-oxathiolane ~"1,, (0.28 g, O, s~ nmol) was dissolved in l,2-dichl.oroethana' (ZO ml), and silylated aytosina ~,,Z, ' (0.2o g, 0.98 :amol) was added at once at room temperature.
The ~oni~ctuure was stirred for 10 ainutes and to it was added .
snCh solution (0.80 ma., 1.A M solution in CB~ch, o.so ' , , ~ ~ . '~. . .., . .: , ; _ ..: , ~ : . ~ . . .. : , . ~ ~ f l~
mmol) dropwise at room temperature. Additional cytosine ~, (0.10 g, 0.39 mural) and SnCl4 salution (0.60 ml) were added in a same manner 1 hour later. After completion of the reaction in 2 hours, the solution was concentrated, and the residue was triturated with triethylamine (2 ml) and subjected to flash chromatography (first with neat EtOAc and then 20% ethanol in EtOAc) to give a tan solid (100% 8 configuration) (0.25 g, 0.54 ~omol, 80%) . ('H -NMR (DMSO-d6): 7.'75 (1H, d, H6) J=7.5 Hz: 7.65-7.35 (lOH, m, aromatic-8,: 7.21 and 7.14 (2H, broad, -8ZH2)t 6.19 (1H, t, H5.) ; 5.57 (iH, d, Hs) : 5.25 (iH, t, H2.) : 3.97 (1H, ' dd, -CHZO) J=3.9 and 11.1 Hz: 3.89 (1H, dd, -CHzO): 3.41 . ~: -(iH, dd, 1H;~) J~4.5 and 11.7 Hzt 3.03 (1H, dd, iH4,) J~?;
0.97 (9H, s, t-Bu)) Silyether ,~, (0.23 g, 0.49 mmol) was dissolved in THF (30 ml), and to it was added n-Bu;NF solution (0.50 ml, 1.0 M solution in TFIF, 0.50 mmol) dropwise at room ~ .
temperature. The mixture was stirred for 1 hour and concentrated under vacuum. The residue was taken np with ethanol/triethylamine (2 ml/1 ml), and subjected to flash chromatography (first with EtOAc, then 20% ethanol~in EtOAc) to afford a white solid yg, in l00% anomeric purity (8CH-i89~ 0.11 g, 0.48 ~mol, 98%), which was further recrystallized from ethanol/CHC13/Hexanes mixture. (1H NMR
(DMSO-db): 7.91 (1H, d, FI6) 3=7.6 Hz: 7.76 and ?.45 (2H, .
broad, -NHZ) : 6.19 (1H, t, H5.) : 5.80 (1H, d, H5) J=7.6 Hz:
5.34 (1H, broad, -OH): 5.17 (iH, t, HZ.): 3.74 (2H, m, -CH20) a 3 . 42 ( 1H, dd, 1F~,. ) J~5 . 6 and 11. 5 HZ ; 3 . 09 ( 1H, dd , 3 0 18~. ) J=4 .5 and 11. 5 Iiz) BCH-189 and its analogs can also be.synthesized ;: ~ . .
by coupling a silylated uracil derivative with y~. w , silylatid uracil derivative ~,,g (1.80 g, ~.aa poly was ' coupled with y,Z C1.73 g, 4.13 ~amol) in 1,2-diohloroethane (50 ml) in the pr~sance of snCh (5.0 ml) as described above in.the the preparation of the cytosine derivative .~,. The reaction was complete after 5 hours. Flash chromatography, first with 40~ EtOAc in hexane and then EtCAc, afforded a white foam 3,g (1.60 g, 3.43 mmol, 83~). , ('H NMEt: 9.39 (1H, broad, -NH) 7.90 (iH, d, ~) J=7.9 Hx: .
7.75-7.35 (IOH, m, aromatic-Hj: 6.33 (iH, dd, Hs.): 5.51 ' .
(1H, d, H3) J=7.9 Hz: 5.23 (1H, t, HZ~); 4.11 (1H, dd, -CHZO) J~3.2 and 11.7 Hz: 3.93 (iH, dd, -CfiZO) ; 3.48 (1H, dd, iH;. ) J=5. 4 and 12 . 2 Hz : 3 .13 ( 1H, dd, 1H,~. ) J=3 . 2 and 12.2 IiZ) The uracil derivative ,ig, can be converted to the cytosine derivative ~,,3. The uracil derivative ~ (0.20 g, . . .~
0.43 mmol) was dissolved'in a mixture of pyridine/dichioroethane (2 ml/10 ml), and the solution . 35 cooled to 0'C. Triflic anhydride ('72 ul, 0.43 mmol) was ~ .
added dropwise at 0'C and the mixture was warmed to room ~ w temperature and stirred for 1 hour. Additional triflic -anhydride (0.50 ~1, 0.30 mmol) was added and the mixture stirred for 1 hour. TIC showed no mobility with EtOAc.
The reaction mixture was then decannulated into a NH3-saturated methanol solution (30 ml) and the mixture was stirred for 12 hours at room temperature. The solution was concentrated, and the residue subjected to flash chromatography to give a tanned foam ~, (0.18 g, 0.39 mmol, 91~), which was identical with the compound obtained from the~cytosine coupling reaction.
F'ig. 3 illustrates the synthesis of 5 methylcytidina and thymidine derivatives of ECH-189. The acetate ~ (0.93 g, 2.23 mmol) in 1,2-dichloroethane (50 ml), was reacted with the silylated thymine derivative y~
(1.o g, 3.70 amolj, and SnCI' solution (4.0 mlj in a manner .
similar to that described !or the preparation of cytosine desivativs y,~. (~8 lit: 8.10 (18, broad, l~) f a.75-'7.30 ,(11R, a, 10 Aromstic.HAs and iNbjf 6.32 (1H, t, S~.) J=5.4 HZ f 5. 28 ( 1~, t, ~. ) Ja~4 . 2 8z f 4 . Ol ( 18, dd, lHs., J=3 . 9 and 11. 4 8$ s 3 . 93 ( 1~, dd, lli~. j J~~4 . 5 and 11. 4 Hz : 3 . 41 "s ,., ..'; ,:~ .. .', w.', ; ',. ~ -_: . ~ . . :.~ ' . . ' . , ' '.. , . ~, ,,~~.~.. . " . . .

1H, dd, 1HZ~ ) J~5 . 4 and 11. ? HZ ~ 3 . 04 ( 1H, dd, , IHZ~ ) Ja5 .'7 and 11.'7 Hz: 1.75 (3H, s, CFI) ; 1.0'7 (9H, s, t-Bu) ) ' The thymine derivative ~$, (0.20 g, 0.42 mmoi) was dissolved in a mixture of pyridine/dichloroethane (2 ml/10 ml), and the solution pooled to 0'C. To it was ~.:
added triflic anhydride (100 ~l, 0.60 mmol) dropwise at ..
O~C, and the mixture was allowed, with continuous stirring, to warm to room temperature. l~lter reaching room temperature, it was stirred for 1 hour. TLC showed no mobility with EtOAa. The reaction mixture was then .
decannulated into the -saturated methanol solution (20 ml), and the mixture stirred for l2 hours at roam temperature. The solution was concentrated, and the - .
residue was subjected:to flash chromatagrahy to give a . v tanned foam ~,g (0.18 g, 0.38 mmol, 90~). ('H NMR: 7.70- '- .
?.30 (laH, m, to Aromatic H's, 1NH and H6)F 6.so (1H, v ' broad, 1NH) t 6. 34 ( 1H, t, H~ ~ l J~ . S Hz t 5. 25 ( iH, t, H4. ) J=3.6 8z: 4.08 (L8, dd, lHs.) J=3.6 arid 11.4 Hz; 3.96 (1H, -' 20 dd, 1Hy ) vT=3 . 6 and 11. 4 HZ ; 3 . 52 ( 18, dd, lHZe ) J~5 . 4 8nd . , 12.3 Hz; 3.09 (iH, dd, 1H2~) J=3.9 and 12.3 Hz~ 1.?2 (3H, .
s, CH3) r' 1.07 (9H, S, t-811) ) Silylether ,~ (0.i8 g, 0.38 mmol) was dissolved in TxF (20 ml), and an n-BuiNF solution (0.50 mI, 1.0 M y solution in TBF, 0.50 mmoi) was added, dropwise, at room temperature. The mixture was stirred for i hour and concentrated under vacuum. The residue was taken up with ethanol/triethylamine (2 ml/1 ml), and subjected to !lash r 3o chromatograptiy (first with 8t0Ac, then 2a~ ethanol in EtOrrc) to afford a. white solid ~Q .(0.09 g, 0.3? mmoi, 9?i), which was father recryatsllizsd from ~ :
ettranol/C8C13/Eexanes mixture to a!lord 82 mg of pure '~
_ aompou~id (89~j . (~ l~iR: (in d6~I~380) : ?~?0 (l8, s, H6) t ~ . .
3 5 ? . 48 and ? .10 ( Z8, broad. N~) i 5 .19 t ig, t , M'. ) J=6 . 5 ~I~ s .
.
. 5 . 3 t ( 18, t, 08) f 5.15 ( 1H, t, 1~,.' J~5 . 4 1~x i 3 . ? 2 ( 2H, m, ..
..:,, .. -;.. ~ . . , : ~ - , . . , - .. . ~ , ~ . . .
.: ~ ;..y.:~:°-':'~. ..: ..~. ...v..-..;.~;:.~ -°.'-: '~ .,:
:y~~_'.w; 'v v; _ ~. . v ;° ' - .; .' : -.. . ' ~ : ~. ~ . : -.. ..
.:..-..~ .::;--... .
~.v:..w:__ ,:v... ,::. , °::.:. °'W y: : ., .:.;~:.v .- .,:
y::.:' :_.,,- ,-.. :~-. _. ,w..
. '..-.'. -.; ~..-. .-..; ,-.....:,:.: ~ :, --~y..:.~-' ~..a, .: ,s....._.:.
~:: ;~ :.~. ~,. ~: v:.- ,y, ' :. . . .... _ . ~:

2H~~) 3.36 (1H, dd,~ IFIz~) J=6.5 and 14.0 Hzi 3.05 (1H, dd, 1HZ~) J=6.5 and 14.0 Hzi 1.85 (3H, s, CH;)) Silylether ,y$ (0.70 g, 1.46 mmol) was dissolved in THF (50 ml), and an n-Bu~NF solution (2 ml, 1.0 M
solution in THF, 2 mmol) was added, dropwiss, at room temperature. The mixture was stirred for 1 hour and concentrated under vacuum. The residue was taken up with ethanol/triethylamine (2 ml/1 ml), and subjected to flash ' chromatography to afford a white solid.~,l (0.33 g, 1.35 mmol, 92~) . (~E NI~t: (in d~-Acetone) : 9.98- (18, broad, NH)t 7.76 (1H, d, H6) J=1.2 Hz: 6.25 (18; t, H~.) J=5.7 Hz;
5.24 (1$, t, ~i~.) J=4.2 HZ: 4.39 (1H, t, OH) 3=5.7 Hz: 3.85 ( 18, dd, 28y ) J=4 . 2 and 5'.7 8z i 3 . 41 ( 1H, dd, 1.AZ~ ) J=5 . 7 'and 12 . 0 HZ : 3 .19 ( lli, dd, l.HZe ) J=5 . 4 alld i2 . 0 $Z i 1. 8 A
(3A. s a Via) ) ~ . .
Fig. 4 illustrates the synthesis of enantiomerically-enriched -189 and its analogs. Allyl butyrate ~ (19.0 g, 148 mmol) was dissolved in CHZClz (400 ml), and ozonized at -78~C. Upon completion of ozonolysis, dimsthyl sulfide (20 ml, 270 mmol, 1.8 eq) was added at -78~C and the mixture was wanasd to room temperature and stirred overnight. The solution was washed with water (100 ml x 2), dried over MgSO4, filtered, concentrated, and distilled under vacuum (70-80~C at 0.5- . .
O.s mm Hg) to give a colorless liquid, (17.0 g, 131 Col, 88~) . (~H lit: 9.59 (1.8, s, H-CA) i 4.66 (2H, s, -CFi~O) ~ 2.42 (28, t, C8aC0) J='7.2 8z t I.7i (2H, sex, -C8Z) 0.97 (3H, t, C83) J=7.2 Hz) (ZR (neat): 2990, 2960, 2900, 1750, 1740, 1460, 1420, 1390, 1280, 1190, 1110, 1060, ; ' .
1020, 990, 880, 800, 760) ' _ Butyryloxyaostaldebyds ~ (15.0 g, 115 mmol) was .
dissolved in tolusns (200 ml) arid mixed with thioglyoolic . .._ acid (B.O~nl, 115 Col). The solution was ratluxtd !or 5 w hours while the resulting water was s~moved with a Dean-Stark trap. Ths solution was cooled to room temperature ~ ~_ and was transferred to a 500 ml saparatory tunnel. The solution was then washed with saturated NaHe~ solution.
These aqueous washing wars extracted with diethyl ether . , (200 ml x 2) to recuperate any crude product from the aqueous layer. The ether extracts were added to the toluene layer and the resulting mixture was washed with water (100 ml x 2), dried over Mg8o4, filtered, concentrated, and distilled under vacuum (°70-80~C at 0.5-o.s mm Hg) to give a colorless oil ~ (19 g, 93 mmol, .
B1~) . (~H Nl~t: 5.65 (1H, dd, &~~ J=5.0 and 1.4 Hz; 4.35 (1H, dd, °CHZO) J~3.2 and 12.2 HZ: 4.29 (1H, dd, -CHx4) J=5.7 and 12.2 HZ: 3.72 (lei, d, -CHZS) J=16.2 FiZ3 3.64 (1H, d, -CHiS: 2.34.(28, t, -CHZCO) J=7.2 HZ; 1.66 (2H, sex, -. 15 CHZ) : 0.95 (3H, t, CH3) J~7.2 Hx) (IFt (neat) : 2980, 2960, .
2900, 1780, 1740, 1460, 1410, 1390, 1350, 1300, 1290, 1260, 1220, 1190, 1110, 1080, 10'70, 1000, 950, 910, 830, w ' 820, 800, 760). .
Pig liver esterase solutian (90 ~l) was added to a buffer solution (pH 9, 100 ml) st room temperature, and the mixture stirred vigorously for 5 minutes. The butyrate ~, (2.8 g, 13.7 mmol) was added, all at once, to ~' ~ .
the esterasefbuffer solution and the mixture was stirred vigorously at room temperature for 2 hours. The reaction ._ - .
mixture was poured into a separatory Funnel. The reaction flask Bias washed with ether (l0 ml) and the washing was ~ -- combined with the reaction mixture in the funnel. The . .
-:. , .
combined mixture was extracted with hexanes three times (100 ml x 3). The three hexane extracts were camb3.ned and ..
dried over MQSO4, filtered, and concentrated to give the -optically active butyrate ,,'fig c1.12 g, 5.48 mmol, 40fi). ~. ,..
Enantiomsric excess was determined by an Nt~t experiment using a Tris(3-heptatluoropropyl-hydroxymethylene)-(*)--camphoratoj europium (==I) derivative as a ohemlaal shift rsagantt this procedure showed approximately 40~
. enrichment for one snantiomer. The remaining aqueous ' 'w :: .:
. ... . . . _ .,,, ,, . : . - . . . . . . - , . _ . . , . . .
': v,. .:.,~ . ~.~..'.~.y:,v~ ..,.;,.: . .: .. -:.,,.: ... ,.:.. ' y. .'.v ~y ..v.;. ~ .. ,~. ~..' ~. . . ~ .~.: _.
.. _. . ~:: ::', y~v. . . . v . . :v: :. . .'.; , . .. .. . . . .., . ' :. ; .
. : . ...- _..
~-..v:. ~w~. ~~ ~.w~.:. S' ' '~~:~.~ ~.' .~-v v.-~= . w.. .:.- . -.:- .w~: . :
.: . y .. . :.:., layer from the reaction was subjected to a continuous extraction with C~iaCl2 for 20 hours. The organic layer was removed from the extraction apparatus, dried over Mgso6, . ' filtered, and concentrated to give an oil (x..24 g), which 5 was shown by NMR analysis to consist of predominately the . ' 2-hydroxymethyl-5-oxo-1,3-oxathiolane ,Z,~ with small amounts of butyric acid and the butyrate ,~. _ .
The lactone ,~, (0.85 g, 4.15 mmol) was dissolved l0 in toluene (30 ml), and the solution cooled to -~e~c.
Dibal-~i solution (9 ml, i.0 M in hexanes, 9 mmol) was added dropwise, while the inside temperature was kept below -70°C throughout. the addition. After the addition was completed, the mixture was stirred for 0.5 hours at -15 78'C. Acetic anhydride (5 ml, 53 mmol) was added and the mixture, With continuous stirring, was allowed to reach room temperature overnight. Water (5 ml) was added to the reaction mixture and the resultant mixture was stirred for 1 hour. MgSO~ (40 g) was then added and the uixture was 20 stirred vigorously for 1 hour at room t~nperature. The mixture was filtered, concentrated, and the residue flash chromatographed with 20~ FstOAc in hexanss to give a colarless liquid ~, (0.41 g, 1.86 mmol, d5~) which was a mixture of anomers at the C-4 position.
The 2-Acetoxymethyl-5-acatoxy-1,3-oxathioiane ,~ ~ ' (0.40 g, 1.82 mmol) was dissolved in 1,2-dichloroethane . :a (4o ml), and to it the silylated cytosine ~,,~, (0.70 g, 2.74 v .
mmol) was added, all at once, at room temperature. The . ...
3o mixture was atirrad for 10 minutes, and to it a snCl; -solution (3.0 ml, 1.0 M solution in CxiCl.Z, 3.0 mmol) was added, dropwise, at room teap~ratur.. Additional SnCl4 . solution (1.0 ml) ryas added after 1 hour. The reaction . '.
was folly by TLC. 'Opon coaoplation of the coupling, the . v solution 'ras c~a~tratad, the x~ssidua vas .fixituratad with ' tristhylamins (Z nl) and snbjactsd to !lash chromatography (first with neat EtOAc then Z0~ ethanol f.n EtOAc) to give a tan solid ~ (0.4Z g, 1.55 mmol, 96~). (~H NMRi 7.73 ( 1I3, d, Iib) J~7 . 5 HZ: 6 . 33 ( 1H, t, H,~~ ) J~4 . 8 IiZ : 5 . 80 ( 1H, d, HS) J=? . 5 HZ ; 4 . 52 ( lli, dd, iHs. ) J~5 . 7 and 12 . 3 HZ
4 . 37 ( 1H, dd, llis~ ) J=3 . 3 and 12 . 3 HZ i 3 . 54 ( 1H, dd, HZ. ) .
J=5.4 and 12.0 Hz: 3.10 (1H, dd, i~i3): Z.11 (3H, s, CH;)) The S'-Acetate of BCH-189 ~, (140 mg. 0.52 mmol) was dissolved in anhydrous methanol (10 ml), and to it was .
added sodium methoxide (110 mg, 2.0 mmol) in one portion. ..
l0 The mixture was stirred at room tnmpsrature until the ~.
hydrolysis was complete. The hydrolysis took about 1 hour, and the reaction was followed by ThC. tOpon completion, the mixture was th:n concentrated, and the .residue taken up with ethanol (2 :al). The ethanol .
solution was subjected to column chromatography using ' "~.~
ethyl acetate first, then 20~ ethanol~in EtOAc~ta affo~d~a white foam (110 mg, 92~), which exhibited an NMR spectrum identical to that of authentic 8CH-189, ~.

Claims

1. A method for preparing a nucleoside composition comprising a 2',3'-dideoxy-3'-thia-pyrimidine nucleoside of the formula that is enantiomerically-enriched at the 4' position, wherein Y is selected from the group consisting of fluoro, chloro, bromo, iodo, alkyl, alkenyl, alkynyl, hydroxyalkyl, carboxyalkyl, thioalkyl, selenoalkyl, phenyl, cycloalkyl, cycloalkenyl, thioaryl, and selenoaryl;

comprising the steps of:

(a) reacting a stereoselective enzyme with a lactone having the formula wherein R is an acyl protecting group, to form enantiomerically enriched 2-hydroxymethyl-5-oxo-1,3-oxathiolane;

(b) reacting the enantiomerically enriched 2-hydroxymethyl-5-oxo-1,3-oxathiolane with a reducing agent and a carboxylic anhydride to form an enantiomerically-enriched

2-acyloxymethyl-5-acyloxy-1,3-oxathiolane;

(c) coupling the enantiomerically-enriched 2-acyloxymethyl-5-acyloxy-1,3-oxathiolane with a silylated pyrimidine base in the presence of SnCl4 to form the .beta.-isomer of a 2',3'-dideoxy-5'-acyloxymethyl-3'-thia-pyrimidine;

(d) deprotecting the 2',3'-dideoxy-5'-acyloxymethyl-3'-thia-pyrimidine to form the 2',3'-dideoxy-5'-hydroxymethyl-3'-thia-pyrimidine.
2. The method of claim 1, wherein the silylated pyrimidine base has the formula:

wherein X is selected from the group consisting of trialkylsilyloxy and trialkylsilylamino;

wherein Y is selected from the group consisting of fluoro, chloro, bromo, iodo, alkyl, alkenyl, alkynyl, hydroxyalkyl, carboxyalkyl, thioalkyl, selenoalkyl, phenyl, cycloalkyl, cycloalkenyl, thioaryl, and selenoaryl; and wherein Z is trialkylsilyl.

3. The method of claim 2, wherein Y is selected from the group consisting of fluoro, chloro, bromo, and iodo.

4. The method of claim 3, wherein Y is fluoro.

5. The method of claim 2, wherein Z is trimethylsilyl or t-butyldiphenylsilyl.

6. The method of claim 2, wherein X is trialkylsilylamino.

7. The method of claim 1, wherein the stereoselective enzyme is selected from the group consisting of pig liver esterase, porcine pancreatic lipase, and subtilisin.

8. The method of claim 7, wherein the stereoselective enzyme is pig liver esterase.

9. The method of claim 1, wherein R is an acyl protecting group of the formula:
wherein R' is an alkyl possessing 5 carbon atoms or less.

10. The method of claim 9, wherein R' is propyl.

11. The method of claim 1, wherein the reducing agent is selected from the group consisting of diisobutylaluminium hydride (DIBAL-H), sodium bis(2-methoxyethoxy) aluminumhydride (RED-AL) and NaBH4.

12. The method of claim 1, wherein the carboxylic anhydride is acetic anhydride.

13. The method of claim 12, wherein the enantiomerically-enriched 2-acyloxymethyl-5-acyloxy-1,3-oxathiolane is 2-acetoxymethyl-5-acetoxy-1,3-oxathiolane.

14. The method of claim 1, wherein the deprotection of step (d) comprises adding sodium methoxide.

15. A method for preparing a nucleoside composition comprising a 2',3'-dideoxy-3'- thia-pyrimidine nucleoside of the formula that is enantiomerically-enriched at the 4' position, wherein Y is selected from the group consisting of fluoro;
comprising the steps of:

(a) reacting a stereoselective enzyme with a lactone having the formula wherein R is a group of the formula:

wherein R' is an alkyl possessing 5 carbon atoms or less, to form enantiomerically enriched 2-hydroxymethyl-5-oxo-1,3-oxathiolane;

(b) reacting the enantiomerically enriched 2-hydroxymethyl-5-oxo-1,3-oxathiolane with a reducing agent and acetic anhydride to form an enantiomerically-enriched 2-acyloxymethyl-5-acyloxy-1,3-oxathiolane;

(c) coupling the enantiomerically-enriched 2-acyloxymethyl-5-acyloxy-1,3-oxathiolane with a silylated 5-fluorocytosine in the presence of SnCl4 to form the .beta.-isomer of a 2',3'-dideoxy-5' acyloxymethyl-3'-thia-5-fluorocytosine;

(d) deprotecting the 2',3'-dideoxy-5'-acyloxymethyl-3'-thia-5-fluorocytosine to form the 2',3'-dideoxy-5'-hydroxymethyl-3'-thia-5-fluorocytosine.