PH26994A - Antiviral nucleosides - Google Patents

Description

A

Antiviral Nucleosides

This application is a divisional of patent application Serial No. 33532 filed March 14, 1986, '

The present invention relates to 3-azido-3"- deoxythymidine, its pharmaceutically arceptahln derivatives and their use in the treatmant or prophy- laxis of human ratroviral infections. ‘ Retroviruses form a sub-group of RNA viruses whieh, in order to replicate, must first 'reverse 10 . transcribe’ the RNA of their genome ioto DNA (transcription' conventionally describes the synthe: sis of RNA from DNA). Once in the form of DNA, the viral genome is incorporated into the host cell genome, allowing it to take full advantage of the host - Is cells transcription/tranglation machinery for the purposes of replication. Once incorporated, the viral

DNA is virtually indistinguishahble from the host's

DNA and, in this state, the virus may persist for as long as the cell lives. As it is virtvally invulne- rable to attack in this form, any treatment must be directed at another state of the life cycle and will, of necessity, have to be continued until all virus- carrying cells have died. . BAD ORIGINAL Yi el 4 ,

HTLV--1 and HTLV--IT are both rotroviruses and are known to be causative agents of leukaemia in man.

HTLV-~I infactions are especially widespread and are responsible for many deaths world-wide each year.

A species of retrnvirng has now been reproducibly isolated from patients with AIDS. While it has been extensively characterized, there is. ns yet, no agreed name for the virus, and it is currently known either as human T-cell lymphotropic virus 1173 (nT1¥ 111),

AlDS-associated retrovirus (ARV), or lymphadenopathy associated virus (LAV), It is anticipat d that the name to ba agresd on internationally is aquired immune defi- ciency virus (ALDV). This virus (refarred to herein ans

AIDV) has been shown preferentially to infect and des- troy T-cella bearing the okr" surface marker and is now generally accepted ns the antiologic agent of AIDS.

The patient progressively loses this set of I~cells, upsetting the overall balance of the immune system, re- ducing his ability to combat other infections, and pre- disposing him to opportunistic infections which frequent- ly prove fatal. Thus, the usual cause of death in AIDS victims is by opportunistic infection, such as pneumonia or vitally induced cancers, and not as a direct result of AIDV infection. - lb - [ .

BAD ORIGINAL 9

: 26d §

Recently, AlLUV has alro been rocovered from other tissue types, including B-cells expressing the 7 marker, macrophages and non-blood asanciated tinsue in the central nervous system (Ul). This latter infection has been discovered in patients expressing classical . AIDS symptoms and is associated with progressive demy- elination. ILwading to waating and such aymptoma as encephalopathy, progressive dysarthria, ataxia and disorientation.

There are at lemat fonr enlinical manifestations “of AIDV Infection. In the initinl ‘carrier! =tate, the only indication of infection is the preaence of anti-

AIDV antibodies in the blood-atream, It ia believed thant such 'carriers' are capable of passing on tha infection defo by blond tranafusion, sexual intercourse or used : syringe needlea, The carrier atate may often naver progress to ‘the second stage characterized by persistent generalised lymphndenopathy (POL). It in currently estimated that about 20% of I'GL patiants rrogress to a more advanced condition known as ‘IDS related complex’ (ARC). Physical aymptoms ansociated with ARC may include general mnlaise, increased temperature and chronie¢ infections. Thin condjtion veuglly propgrnsses to the final, fatal AID5 condition, when the patient completely loses ths ability to fight inféction. ) -5- BAD ORIGINAL 9

L

Co | | 6A

The existence of thess human retroviruses and others has only recently been recognised and, as tho dideases with which they are linked sre of a life- threatening nature, there exists an urgent naed to develop ways to combat these viruses.

Various drugs have now been proposed an ‘cures’ for AIDS. These include antimoniotungstate, suramin, ribavirin and $soprinoaine, which are either somewhat toxic or have ahown no matked anti-retroviral activity.

As the AIDV genome is incorporated into the host cell

DNA after infection and is virtually invulnerable to * attack in this atate, it will peraist as long as the host cell survives, causing new infection in the mean- time. Thus, any treatment of AIDS would have to be for an extended period, possibly life, requiring substances with an acceptable toxicity.

LL Ce Reporta have described thn testing of compounds. re against various retroviruses, for example, Friend lLeukaa- mia Virus (FLV), a murine retrovivuas. For instance

Krieg st al. (Fxp. Cell Res., 116 (1978) 21-29) found 3'epgzido-3'-deoxythymidine to be active ngainat FLV in in vitro experiments, and Ustertag et al. (Proc. Hat.

Acad. Sei. (1974) 71, 1980-85) atated that, oa the basis : of antiviral activity related to FLV and a lack of - 6 - : . BAD ORIGINAL 9 oo | 2aq ‘cellular toxicity, 3'—azida-3'-dacxythyniding "might favourably replace hromodeosyuridine for medical treatment of diseassg cauand hy INA viruses".

De Clera et al. (Biochem, Fharm, (1930) 272,, 1849- 1851) eatablishad, six vears latar, that 3°. deoxythymidine had no appreciable activity any viruses uged in thelr tests, including vaccinia,

HEVI and vericmlla zoster virns (VZV). Clinski et al. (J. Orn. Chem. (1973), 38, 4199-41305) discloacs certain derivatives of Jeuride-3' -Aanvythymidline (Infra) and thelr ability to hiock wamealian exori- bonuclease antivity. . - We have now discovered that 3'-arido-3'-deoxy- thymidine has a surprisingly potent activity against human retrovirnsms, «with a particularly high activity against AIDV as demonstrated by the a<parimental data veferred to helow. Such activity -anders the _ codpound useful in the theraphy of human retroviral "infections.

Thus, in a first aspect of the prceant invantion, there ig provided a compound of fornuls (1): . Lo oo TACINAL dP

Babs Or

0

A. OH

AN +

I Ae 0” >%

Ho LO - \__./ m, (1. e. A amido carat hem bd ban Coy 2 - vharmaoantine) ty acaba be Am fogb fae

Fhaoyaef for vies bp IP Fpsen meat ar prophylavin of hops ratyay ive Infect inne )

B the compo of terms re ard He pharmacentionl lv noosotabys ola vrata ag Ape herauf te, ve ferred te np tha Carnie according to the Invention ’ | Bel tvity of A -mnlde 3 cdaovethvind od tne @ aaninat hman yetrovieasen hop Boor artabl ished . bri way dean in v)trr SELEY Aviat Foy \ axpmple, infecting af the HR hvman

Iymphabtantold rell- line be ATHY |p nf foctively i prevented be corcebrations of A" nzide-as 16 deoxy Lhymldine ap low se 6.010 meg sm vp te 20 oo benes after infeat ion. AIDY infant icn o § HIT om .

BAD ORIGINAL J oo | 2qq( i himan lvymphoblaagtoid colle, PHA - emtimnlated ~ a white hlood nelle and eulinred periphsral blood Fymphonvien fe aloo prevent «rl at eimilarly low concaptrationge In addition, 100. 6 day challange axpaviments nping up bo BOA ATDY virions par cal) and alonad T4, fetannnp : apecifi,, T-helper Jwvemphocvtas, slowed no decreass In calle trested with 2 -asido-3 - deoxythvmidine, while untreated aells had 19 decreapged h-fold. COytopathie effacte were alec completely blaockad in the seme amll-1ine trannformad by HTLY-T and puper-infanted with

ATDV.,

Other studias uring purified AINY ravarage 16 transcriptase have shaun that the activity of this enavme ie blocked by the triphosphate of dA ~anido-3 -deoxythvmidine by a competitive inhibition mechanlem.

Fhaea TT olinical trisle have ahoun that 3°.

A" azido~-3 ~daonxythymidine is aapahle of orogeing " . the blood/brain harrier in clinteally effective quanntitiece. This property ie both

BAD ORIGINAL 9 a I) 53 ot ‘ “py ; Vo . : ii sy io . E s ‘ MN £2 i: BE a IB

NE angst and valuable for the bEreatwmeal and 0 3 KY. - . . . (I

Fi prophylaxis of CHS infections caused by human ‘4 _. rebroviiinusss. iH yr i i 3 kl:

HE il

A hs The ability of 3 -asido-3 -deoxvthvmidine Bd if 8

SS: . Ln . IR ‘ b to wodify the course of retrovirus induced fH ge i J + 3 malignancy has bean demonstrated in a monse 4 a ae

Ly modal, whareby administration of 3 -azido-3 #

REY] - Fhe

Lig deodyvthymidine prevented splenomegaly caused by IE

SR LE

4d : intravenously administered HKauscher Murine iB a. if » ; i ; i 13 Leukaemia Yirue, the marine eguivalent of HTLV - iE

I ” | I) yor : aq A , hy I. lin Turthor experiments, 37 asido-37- | § - .

Y % decsvthyvmlidine has been shown to inhibit the ja a: mn vitro replication of HTLV-T at ooncentrations Do 4 3 as low as 9.8 meg ml. Ve bg : i : i ne =. 15 Thus, in a further, preferred aspect of the Eb bh , . . it! ; i present. invention, there is provided the use of 2 x a componind according to the Invention in the & 3 manufacture of a medicament for the treatment ? : or prophylaxis of human refrovirng infections. | £ ® 2a The praesent invention farther provides a FB

BR. ¥ ; 4 method for the treatment ov prophyviasis of AIRS ii x 4 p 4 ji! 1 ad 3 ‘WR 5 4

B! FS

M 2 y ts i 1 - 1 i i]

A i 4 nA i, I a , AL id » BAD ORIGINAL oy i “3 a.

Eda: St ER ta ERI Copmd aA ee Wh BRE Lo. Cle . . o

Pe ene. ER ET TERMS RTT J pe SE

HAL ce A we To : gt . SP NI Bik

Are TY ie : N Cy

ER in » human eubjant whinh cowprirer i - | aduwinietering te the sald human enbject an : affective amount of a cowpound according to the

IEEE invention. b Tha present Invention also includes a . method for the treatment or propheinxie of : infections oauged hy retroviruses ‘nv a human oe Co aubjgzct, which comprises adninisterivg to the : N, a

Co sald human aAubject an effectyve antlvira) oo 109 apount of a nompound according to the ay invention. : i . . . A .

A ~~ The praement inventlon further provides a , compound acaording to the invention for vee in : oH ee mmm phe treatment of prophylaxis of AIDS cr oa virus

Sh : ya : : 18 infection ans identified above. er . 0 Examples of human ratroviruy infections :

Ty _ ®hieh may be treatad or prevented Jon accordsnce

Ch with: the pressnnt fuventlon fncinde T-cell

To lymphotrople retrovirvess (HTLV). especially

Ce 20 HTLV-I, HTLV-11 and ALDV (HTLV-II1). . 1 - | - BAD ORIGINAL 37

. i - wee HERE SE ek + gd i;

Si | . - Olinieal conditions that may be treated or ’ prevented in accordance with the tnvention oe | include AIDS, AIDS-related complex and HTLV-T positive leuksemia and lymphoma. Suitable . Co 6 patients for treatment alec include thore - having antibodies to AIDV, AIDV CNS infections,

CL - PAL and ARC. " By "a pharmaceutically acceptable i - derivative” Js meant any charmaceutically i. 10 acceptable galt, ester, or salt of such ester, ’ z : or any othar compound which, upon oo | administration to a human subject. is capable oo of providing (directly or indirectly) a- - azido-3’ -deoxythymidine or an anti-retrovirally 16 avtive metabolite or residus thereof. An

Sn v example of a non-eater compound Is the

MA : derivative wherein the B-C- and 2-C-atoms nt are linked by an oxygen atom to form an anhydre

Ea - group. { : .

B ; 20 + Preferred esters of the compound of formula Ce | (1 include carboxylic acid estara in which the .

Cd So no - 12 - : ; oo BAD ORIGINAL 9 = ne oo me bh eh Coa . CL . . o

Wa orn: CE TT wg ask

LT — nolety of the ator grouping ia aalected fram strataht or branched chain alkyl, ] } Alkoryalkyl (e.g. wathaxvymethyl), aralkyl (ag. oo benny 1), arvicwyrslkel (eg. phonavymathvl), oo Bb aryl ta.g. vhenvl «ptionsliy substituted by oo halogen, Cy.q »leyl or Cy, alkoxy); sulphonate niutore Anh Be Alka d= or ) aralky lan) phony! Cie og wathaneauliphions yy ad - . nono df, or tri-phogphsta esters, With - 1a regs to tha shove degsovibed cplors, unlsen abherulne ppenitied apy wlhvl peiablos present in swh enters sbeontngeonsly contain {Lo 18 : carbon atom, poartionlesrly | toe 4 earbon sloms. ] Any. muy! molaty preagent din gnch enters 3 : 15 adysntageoue iy Acmpriean nv vhenyl gronp, Any oo referen-e te any of the hove cowmpoonde salen oe oo ineludes & cafacepers tas pharmaceutically a. at nocsptuhle salt Lhareaf.

Co Rxperimente have phown that 37 -naldo-3°- )

Ay 20 deoxythymidine ie cemyartad, ‘ig vitro, by the : action of asl line earymes Into Lhe br- E

Fr : manopharphato The mormphosphste 16 then

Be | R further phosphorylated by other enaymes to form : \ : i . ) EB \

So : - 13 , - : “ Co co BAD ORIGINAL 9 :

Wer CWT ga

Vo pith Ce Eo. Sagrp J 7 : HEE Ch | : we : awl

Ea : LY

CE : ‘

Lh : Lo . - eo. the triphoephnte via the divhosphate, and . pa oo other stndlos have demonebrated that 11 is the ] i } triphosphate form of 4° anido-3 -deoxythywidine

So which irs balieved Lo be effective chain

Cv : b terminator In the reveres hranacription of " ~ AIDV, as svidenced by ita mffect on avian : d ’ myelobl satosin virus ane) Holonay murine oo : lvekaemia virus. This form slsu fuhibits AIDY

Nr reverse Lransariptase Lp vitro whileh having n

BR o 11%] negligible affect on tmman DIA polymarase 5 . | aativity. : y

Le ) _Bxemplas of pharmacentically a i asceptable ealte of the aompound of formula (1)

Co and 1ts pharmacent.iocal ly acceptable darivativer 16 tnolude bare nalts. eg derived from an ee appropriate haga, such ae alkall meatal (e.&.

Sl ~ sodium) . alkallue earth metal (e.g. mognosium)

RE © anlta., ammonium and WX', (wherein X is Cy 4

Sa alkyl) co Bb | 20 "The prasent invention thus further provides the nove) pharmaceutioally scraptable i dexivativen of the compound of formula (1, . ) ~ . oo - 14 - or : | AD ORIGINAL 9

ER] Lm

. . . . oo Cua a. wr man ® aes em PET fe Al vi fi : . i Fyne ’ ’ ae A Lon “ wt > ! ’ } : a « he rv ie a Toa A y ). pi AE

RES doe ET eto ee co - SUS . 4 : CRA AE gs 0 CR gan i hl . . Ce - ee TY & ih E LE

FEU Te Co CE TIN or gy ge wh fos ' “ TA RES EE ade oo Sh A : Neg peste. ne oy Ta : : ather than the tollowing & dertealives, osmely ; monophoiphatss, diacd ium mencphios pata, 2- aymnoe thy L nonophorphate triphornhinte P-

Co tolnen: sulvhonete, acatate tyripherrTasthyl

Po b snd’ me bhnuocolphonste deyiyativer ar where the

S741 je lioked to sm Turther eneleciide or nuglace id: dapioating,

CBpaeaifie axsing las of pharmacentically . 4 7 acoepinhle doerivabtives of the compound of ep 19 formula (1) bhal map be used An accordance with : no tha proecent Invention tuoinds the o~nosodium “i,

Ce sald: and the following b- eatera:

Te monophosphate digcdium ronophosphate

SE dighoerhate: triphosphate: scstate: 3.mathyl- 16 butyrate, catanoata: palmitata: 2 -ahlore con bengoats; bhansmnoate: 4-mothyl bansoaim: hydrogen : / succinate: pivalats: and moaylate, - :

Le ol : a BART IN-3 Aan vihrpmt {He Cr s rharascanticonllv arraptaiie dertvailes (thepscf soko 20 (hereafter reforynd to AR the agtive .

BEE oy Ey fd ingredient). may ha adminipnterad to baanps for ir } : . prophylaxis oy treatment of retroviral : . HE ' ‘., . . ol : oo | - 16 : i | : BAD ORIGINAL I) ' :

hoo Re CEL ten ny AES Lo ‘ vy . Cr . JEN FA oo ds Chi ET ins

Bidet Uo 0 Te Ee eR

RAN LE ip AYR, SAREE hr aT ee ee TE : EN BE po so Re in] a | Rog, LE an AR : , . Juha " . ACER Lin

PEE CL REN oo of

EAR Te Con et

OE i | Le "a . : : 3 oT infections by any aAvitable ronte including oral, rectal, mnenl, topioal (including buccal * and sublingual). vaginal snd parenteral (inc inding guhoutisnaons, intramasonlar, 5 Antravenone and tutradarmsl). It will be . appreciated that the preferred reonte will vary

Co Co With the condition and sge of the raciplent, the nature of the infection snd the chossn . NE active ingredient, " \ . co \ 12 ~ In genera) a suitable dose will be in the

Sree t range of 3.9 to 120 mz par kilogram body welght nd n of the patient per day, preferably in the range

Ce of 8 to 9@ mg per kilogram body weight per day ol Cost !

Vi \ a :

A } and most preferably in the ranga 15 to 60 ng ;

Col . 16 per kilogram body walght per day. The desired

Che dose is preferably presented as tw. threa, ny Th, - oo

CL four, five. six or more sub doses administered ;

Tr at appropriate intervals throughont™ the day. pw

Lhe These mub-doses may be administered in unit od 20 dosage forms, for example, containing 10 to : sh 1602 wg, preferably 20 to 1000 mg. and most

Ci Ce Lo wl preferably 5 to 780 mg of active ingredient iu) ~ : : oo - ; chai el - 16 -

Co oo | GINAL a ; os Bh de Ek LE : Tv, WL i amas = ’ Cay i CL

TREE ve * CL

TRL Te ped

EL . go

Chri af : :

Cpa

Ei cr ) : hl per unil doenga form. i. to CH Expariments with 3 -azido-3 -deoaythvmidine suggest that a Mose should be administered to achievne peal plasma concentrations of the } b active compound of from about 1 to about 7H uM, pretershly shout 2 to HQ att, most prafsrably : about 3 to shout 3P uM, Thie pay be achieved, for examples, Ly the inkravencus infecticn of a “~ 3.1 to BY solution of the active Lgyrredlent, vy 19 optionally in eatine. or orally siwminietered an

EES a holue pentnaining abont 3 to about 1667 mg/kg

Cae : of tha active ingredtfent, Pe=agirahlie blood

Co levels may he maintained hy ao continuous . - ’ infpticn te provida aboul, 3.61 te atont 6.0 £0 Co 1b ma /Xg/ hour or by . Jntermittent tnfusionm

Ll . Co containing about 9.4 to abont 15 mz/ke of

SUE Ca :

EEE the mative ingradient

EE . ‘While it is possible for the active ingredient to bea sdministered alon2 it le vo 29 preferable to presant it as a phevmsosutleal . formulation. The formulatione of the present

Co invention compriane at lasst ona active : - 18 - wa 0 oRiCW fas - BA bo y | Ss -

SRNR oh ie ash RRR AT oe BAY ee RR Th RAE Ta a eh al sir

BAR rr Tn TTL Sn RETR) Arr SET ae RERTR Se e A e R vd ho LAR hae i,

Tis LE IEE wk CE cone Ea ne Lt {ota Wh 2 i Ser

Hanan Bo flee « Co AEE gg

Eh CRE 3 : CNR oe dy 2 dnl i, : : Ay Re] rn wey SR RR Cg

CA T pA - Co Ag

Ra a ; } ‘ . i Cx E . oo he SR . : or ~ belle } . 4 . Co i" Ca we

Spr Cg cn TL .

Co Formulations of the present invention 4 a auitahle for oral administration may’ be

SL Co presented as diecrete imite much an capsules, . cathets - or tablets esch containing a

Lo 5 predetermined amount of the active Ingredient; , . As’ a powder or granules: ag a soluticn or a : . LE . ’ : { ’ sudPena Lon in an aqueous or non-aguecne liquid; : or: ans an oil-in-water liquid emulcion as a . . bolus elactuary or posta. Oral formulations = i pa to ’ 1¥ may further include olher agents conventioual -

Soh. x 3 oo ing, the art, such as suesteners, flavouring

Re :

Sie ’ : ght and thickenara. -. Co . KS or ' z 5,

GT Co A tablat may be made by compression or

Rr © moulding, optionally with one or more accessory

Fa Ch - cued

Ke » Coat :

CE Wo 1b ingrédients. Comprampsed tahletr may ~~ be

Phe J Ey : :

RSA Shenv, * , fob prepared by ocmpressing In a sultabla machina . Lo cea Hs \ fd the active ingredient 'n ap free-flowing form

Se sugh as a powder or granules, optionally mixed oh \ with a binder (e.g. povidone, gelatin,

Sr Vo Fei

FE i \ T 28 hydeoxypropylinethyl calluloce). lubricant,

Cod CN el Co

Shiver , . inert diluent, preservative, disintegrant (e.q. :

Le, , 4

IE \ Lot . nT sodium starch glyeollate, vrons- Linked

Pe : \ RCS cred \ 5 2 ! \ } i ' . . .

Ea ‘ Li rN

Wh RE

Sa x,

ET aE ee

ETE ot : r-

Coa | 5 le BAD ORIGINAL J

EE I. A

ER a mnt ee a AE ne lid peat SA he eC iow Mo nF eh ea saws se

SRI a Re TAA ABORT Flv oli Ee rT Sn a IER a © aN PR 140 - Ei A ih rift. 3 Sante Sa at co : wo . A an Vi. \ i SG LA» rt ye ame RR lta el HR : : CER RTs Tan

SUERTE LE TM Haw

SE Ce Cee ES RRR

CT marr mr re . 7 we g - : povidone, «rosa-linked sodium oarboxyrethyl cellulore), snrfaca-antive or diapersing agent. : a Monlded tablets may he made by moulding in =a

Co suitable machina a mixture, of the powdered

PH B compound mnietenad with an inert liquid

CL diluent. The tablets may optionally be coated

By or .soored snd may be formulnted sc as to

LL provide alow or contvolled ralease of Lhe

Pel active ingredient therein uvelng, for example, oo 1A hydroxvpropylmethvlcellulose in varying proportions to provide the desired release -

L profile. 0 A ; . Formulations euitable for topiasl : administration in tha mouth Include lonenges ; 16 comprieing the active ingredient in a flavoured

Chere ; , basis, usually sucroge and acacia or

Ch : tragacanth; pastillen comprising the active

Lo ingradient in an inert basis such as gelatin i and" glycerin, or sucrose and acacia; and

SA . 20 moitthnashen comprising the active ingredient in

Tro a suitable liquid carrler. . oo Ce » } . : Nasa wea

: po : whee een RESET NE ed

Fe Riis sn. . : Co wi) SR EE A or . 3 . whe Lh g ob , + Tha 0 on ¢ Re SY

Pee cL Eh 18 - SU | Ses Shoat me Cohn HS . Ce TR i SR

Tagan sy WT REE CE ee

Role, oo es | Tae RR

BAS SO : oo . ; ein Sr | | i

Co.

Fors lations for ractal adminiabvation way be presented pa os eappopttory with a endtable base cawbricing for eeampls nocon butter or a salioviate, b > Warmulatione suitable for vaginal : oo adminietration may he prepontad ss teogarfes,

EX tampons. cvasma cole, pacten, foam op apray . Ce to formulation onntaining In sdditien t3 the eo

So ANE " . co avtlve Ingradlant ench ne rarvrisrn ap Are known oh . ’ ip in The art to he ap ropriaste oo 7 Fovsulat bong auitahte for parenteral adminletration inolnde aqueonag and nan. aqueous } lsotonie mtar)la intection molntions which aay : : aontatn anti-oxidante, buffers, bactariostats ne 18 and. anlntee which vender the formulation

To So frdtonle with the hlpod of the intendad :

HE recipient; and squocne and non-agueoue sterile

LL * suspeheione which may innlude suspending agents . CL Aa. .

RTE sod: thickening agents The formulations may be :

Re, Cie Cy

Gg 7 TL 20 presented in nnit-dose or multi-dore gealed - we Co 1- containers, for exampls, ampoules and vials,

CERT cn . i kN es . me Sow

Ln . Le . a i

CL ST

Co ie BAD ORIGINAL 9) . . Lh | | -

Wepnoo pains 0, rR Lad an ath Lo To TEE a eq AR al 43 JOE mR

SBR CT ERR Te DE Be em el et tn LE kha oR ginger gh t -Hetub REIT Pr

FER Cb CEH . ot Ving gl NU fine

ST Cn ny SE THOR BE

Bee Co Ch : SE CARE Ee ET

RE po MPL oe LE LENE a TARE FL

SEs NS CTR he Tr CTT i Lo . “ds ;

LTA : ’ ea p ) ’ snd may ha Atoread in A franme dried

Co {lyophiliaad) aondition requiring only the

Lo addition of the sterile liquid carrier, for

Co oxawple water for injections. immediately prior ,

Ty 6 to use. RExtemporansous inlection solutions and - oo guspanalona ~~ may ba prepared from eterile powders, granules snd tablets of the kind

CL \ : previously depcribsd. : oo uo CPrefarred unit dosage formulations are

EE 10 those containing a daily dome or unit, daily

Yeo yt .

Lo sub~dose, as hereln above racited, or an

FE appropriate fraction thareof, of an aotlve hn eo co Ingredient. : . The adminirtered ingredients may also be

LL 16 used fin therapy in coniunation with other pu x os medicaments such BR CP {T2-hydroxy=1-

CL } . (hydroxymethyl )ethoxy Imethy1lguanine. B-(2-

SE hydroxyethoxymethyl)guanine (acyclovir), 2- a amino-9-(2-hydroxyethoxymethyl)purine, : iL ) 20 Interferon, s.g., d -intevfaron, intarleukin

Sr Si -

ERA - II, and phosphonoformate (Foscarnet) or in ub K conjunction with other imonne modulating -

Ca mn - 21 . ih SE

CL je BAD ORIGINAL 9

L . Le

- i

A pis HN : v5 1 3 Be ' 1. 2 k: : : i i! = 3 : 2

B ll -

Hl a therapy including bone asrrow or lvaphocyte i = 1 : 1 i tragsploats or medications such as lovamiool or !

TE. i ‘. thymosin which serve to iaoresase | yuiphooyvite i : ¢ mute rs and/or fanctlon ss in appropriate. | g i. | i

H 7 3 i 5 : 3 It shondd be understood that in addition to Ni

BR the dopredicots particolacly mentioned shove i

LL X . 3 M i

F the fTormoalations of thls invention may irc) ode a] - 0

Ml athe npenla convenhbionsl in ble arth wf i 1 !

Bb Ri x Foret ot boa hl

E : iu Thee Gompenpd of formala (1 apd its f Z

BR ol k4 i . V . . A iy 4 i Fharmacozint foal Yo oaoeeptables derivati VES lay les i. b 3 [1d i Co oo [i

I prepared do conventional manner, for example as n- 4 I k - dasoribad in Lhe following references, or by I J ‘BB melhodis analogous thersto: JOR. Horwits ot al. i.

Li IE . Re 156 Jd Chee hein 29, (daly 1964) HATE T8 IH. 55 Bata cb a), J Gry, Chew. 43 (15) (1978) | 1 8 i BO44 BAH KLAL Watanabe el al at., J. Opg. 1 [| LE i: i ¥ Chem. 40, A274 (L682); and H.F. Glinski et x. ii qa alo, J Chem. Bool Chew. Common. , 915 (1976), il 1.

RB

ED oo xX 24 as well as the processes dexoribed in the i 1 Esamplen, f ; i fl @ y at j i 3 1 3

A x

IES he os = Sun 3 : 4 Foe 5 a BAD ORIGINAL 9 i :

Oy a

Sb Ye LL Co ER Ls a : AA ne The SAE gop ch a a CEE Tn TR

Grae re he CAME SE ES lg, He An

TEAR A ave . ee . ERLE Beil 4 I Sop mE Cah e SR ja 99 “L

AAT Lo CER TREY pe

Sih i Ce

SRE oo : SE

Co : . The present Invention further includes a : procese for the prepavation of a compound of formula (I) snd pharmaceutically acceptable Co

Co derivatives theraof which comprises

Cot - 5 © (A) renching a compound of formula: . ' . oe | .. f on - } :

Lv t h A

Coe . SL RO 5 ~ . ) | “ | . \ J —— : . -

Co - Clg Ce “ Ca ils , . .

RT ] Sage . Cu ' noo

Cas el) Co i

STE EEE SE :

Sohn dat

Lh eo SU (Whereln M represents a prenuracr group of -

EN a :

SE | tha 3 -uaido group) or a derivative (e.g. #n

Sones . ] . CR . .

Lov estbr or galt) thereof, with an agent or under

Ten : ‘ a 2

Cael 10 . cofilitions serving. to. convert the maid : a TA " °

TL. . Shel ’ ; 7 eT pregureor group into the desired nzldo group; - n ; Co or 3

N a - - . A : N

Lo Co Be - 20 -

BO . oC

J nl pr = ’ : ; BAD ORIGINAL ‘ )

£ idk) py Hi Loa Gi Laat nh . Wt 0d EC SL Qo i - ar Lb ag: Ey Li 4 i N et panty oa La een Lem EE SPN TR A. Ti

Bsc eR en UFR il lee Ag oink aE oR Sond TT no wo Eel RH, CT

SE Cees : : : : Ch A ccm i FE

TER hid GE € ix : Nar Clay

TUR tgs ESE Ce Mig)

SE LT Ca TW Tt 0 ATER 4 rr brag de Si mamaria igi” - , or at Tg . - :

STARE od Coen . Tet

SE ou Sy Dare } wo Pale : TU EE

IRR a C

CRE Fb : ke "

Te < : ‘ Sh . : - . ast RB CO .

Lips oe a) ‘

Th LL :

RE o ov on! . .

Ce (B) rascting » nompond of fovmnln:

Cl : LL . 1 | ’ . Pe } ) oo Pl A (111) :

Co R

Pp o

Co j My r Co twharein BR orepresents a precureor group for wy fT : the hydroxy pronp, or for a phermscentioally A

CU ; b acdeptable dJdarivestive group theraef) with on SRA - Ce ’ a ’ yy

CE ! agent or wader conditions serving to convert SL

Coy , } | Re : : , ee t

AoA a Cy en oT FH Ce thg anid precursor group into the corresponding : i

SIRI :- dedired group; ov oo ca ' ! Tore [SE “. or eg . . a ; © Ce 4

T. i ii Sa SEC) reacting a compound of formula Coe Sr oe di ; } Lo CN ’ . Ce at Co 0 i» a

Soh . - FR Co cl CL Loi

A go A 7 on or ‘ °

LE J ~ 3

Lhe a . } L. - j . Pa , } \. 5 ' oa ‘ -, . 4 . [EIR Ta ’ Se - : 4 = ¢ TF

Cpe at } ie St . coe - i pr

Coe Se - 94 - BAD ORIGINAL 9

Ce - A bei - i ~ . Ga } ps ps .

Ce ! . . i ? . ,

¥ — — —_—

ER cibpat Test : crag ne : Carpe eas

SER beet UO pn Tie Pl dT CL We THER eee ye EHR CE pen “a

EIRENE Se en SC ve SETI me et Tay ee GEER ei ig Cg ol LE E& | EE Tee A Co EER rat

SN Shr Fr ion CSTR ‘ i ) ” 2 0). MeO

Plena Leon I « AH ah fet tPF Qn

EERE RR : : TOV aa Negignk OF 200

PIA NEARE ft Cpl : © oe [rn

Cea RETA le JA . . Ek . Sel a AR yt del Cah et Rd i 2h IE ET SHE

Sj Bra g CUR re

CE et EA - B CFE

FE = | f ] 5 Sl Col } ,, . i fo . or functional equivalent thersof, with a

Cy compound amerving Lo introduce the dasired :

Ld ribofurancayl ring abt the l-position of the

Be : compound of formula (IV); or ; i 6 (D) vesoting a componnd of formals Cae ol 5 = 1 on, t oN . oo

Cl ~ : - 2 opr - : 0 (vn) : . ~. 4 CE oo wk - a : : eh SES He oo fans 5 EO pi : BE )

TEM) nd 1 Lc i

EE ne : (ahérein R' is hydroxy or R as defined above), :

FRR Te Wo rio ; K .

PERege TL aT CT : » :

Baily . with an agent or under ennditions serving. to hE EE Lo ob To convert the sald compound into a compound

Er Eh :

Cd 10 according to the invention; oo and thereafter, ov gimtltanecusly therewith, effecting one or more of the following optional oo conversions: . 2 . . o he.

Ce Cig) Cl . oS Ce ae . Ol

Co CL oe . am - 26 -

CT ' ’ ST :

CE i : BAD ORIGINAL 0, :

Lo CE | :

PE Te 8 - So 4 . #

& C —_— —_—

Babine. afin ts TE ae cea he RRL hie a im i EE + REE GLE a et eee eb oe SOS

Een ER : CTT EE RRA Be

Fo A : a age FE HE TR a

Hee ge Ge « ose 74 SR RAE

BAL oA nd Tila i sep ol SA RELA sed, co

TERA . ve ’ ’ Dorr 5

Gk : : oo fi : (1) when 3° —agido-3 deoxythymidine in

Cl formed, converting it into a pharmaceutically

Ee acceptable derivative thereof, - . ’ \ 4 oo ' . \ (11) when a pharmacentically ancaeptable b derivative of 3° -anido-3" -deoxythymidine in formed. converting the ssid darivative into the ov Nr compound of the formula (I), or A At fferent

SU derivative thereof.

Co a In the above-degaribsad process acrording to

SER 10 * the invention, 1t will be appreciated that the

Co] ce precursor compounds of formulae (11) and (111).

Co as’ well am the above-mentioned agents and ney conditione, will be selected from those that

JRE i are known in the art of nucleoside aynthetic

Co KA - Wb . ‘

BLE 15 chemistry. Examples of such conversion ot procedures are . described hevainsfter for

Lg guidance end it will be understood that they : hg oan be modified in conventional manner

Ly depending on the desired compound of formula

Clk 20 (I). In particular, where a conversion ls

CE 5 :

Ce described which would otherwise reaesnlt in the oo Se | : oo v —~ . Ll " . rd + a. 7 oo

CE

CHT i So if a - 26 - r— | :

Lf Tr »

Ch Co BAD ORIGINAL &

& ATR EE TRCN Le RE Rn, oS EY wel Lm Sa SERRE ee wieeel He i EATEN

NEAR ET rr ed TT Sade : : . or Co eR ST ToL EE SY ee

ASE TL nT EH ge a Co yu ’ CRTERE + Wig PARI

SARE a SAW , Cy . fxg] AIO, CL ENE

Co a CHE or | Hi hk 27 Tele

SARA Bo : Ce wn AEST Te ean er rn Son EL ., 0 “i . oA Hl og ey - Co : Ta En ! rh.

En RA . eo : nel

RE : Wr ve

Chea . .- vo * . ’ ’ ) Ct undesired venctlion of labile pronps then gueh : ) . Fronpe may be protected in comvenl Tons manner, ‘ uith enhecquent, removal of the protecting - groupe after completion af the conversion, se 5 The. In proceae (Ay, the prone MH in tha : : compponnd of formless (11) maw renragoagt | for

SL axamp lo, 0 hoa bogen Ger chlorine hudpary or oo : sprains cn phony Toys ep

Ch _ Frifliaoacmet he lanlphonyYoey } ® . - 1a mebhareaniphonuloxy or portale galphanyloxy

Eo radios ) ‘ © Fer 1 he reparat bone of the copay ced of

Lon formula { 1) . a ono wid af fermala | i Y } i [¢]

Cn which the gronp Mode = halogen (ea obhlors) i 16 group in the threo ~oanficnration Cin which the

Cy 5” hydroxy ir advantageously protectsd, ag with ry AU : a tritvl group) may ba treatsd far example with i a Lo

Nip Lithium ov sodium azlde. The © threo halogen

THe Co 3

ST (eg chlorine) rtarting material may he obtained wT . : Geo ~ wel 20 fop example by reaction of the ocarrverponding .

Aaa So Se

Tepe ‘ 3 «arythro hydroxy compound with for example ne i ' triphenylphorphine and tetrachloride, or che alternatively by treatment with crgancentphonyl

Cie pe g oo oT PL LC

Bi . BAD ORIGINAL Ol}

Ce ) -

b

Bato afb LORE TL Te Ee i

Leta] Son EH Fue & HS Lo CL Lh 3 ENE 3 17 tt Av ; 5

EIR TE el Se i Coad cL Reet Cy he

FEOF Sh REY : -« : : oa BCR

EEL gbeca vl, el : Laet C Tne ls Na | RAE appeal Lr co a . To Mees A ¥

Sp es Cn Cem 1

BEET LT - aS

Hala : LE pT

BET . . - . . = - nr bo | oo ' 0 0 ; ~ " fa, ote

Lo CT hE

Lo Co . ’ : Pt

FEA | SE a . . ’ ' 1 i halide (eg trifiuoramsthapesnbplcoy) chlarvide) il to form a corresponding erythro !

Lo Lo . organcnn phony boxy compenpd ubiceh ie then ! 2 LL . halogen on teed Alternatively a 2 theso hydroxy

EE ’ ' ’

So 5 compornd ab formals (LT may be teestad,o tor

EE example uith tripbanvivhonphin., carbon : . t } Ce batyrabeomide and Pithiom azide Go dorm Lhe : i , Lt r . . ’ : !

SE corresponding NT -ery Lhro an ela compound. : ’ ! : Ln ’ i

Lo ; Ramoval of any protecbiygs ppanp peas then pubgeguent Ty be ef Focted oer ain hers

Cot . : oh . ‘ nd te i With regard Lo process (BY, K way vopregent

Cl sa , : : . a Probes bad hudpoxy poranp ooo. ary epter i

EE - i a aroun nie of Lhe type referred boo sbeve dn : EE - :

Co UE i - ’ srtiealarie wralb ory, “i cl Co relation boo farmnds (1) partial ! ALORY

ER ;- 15 DE an ellen weonp such wn tiialke inllyloxy i

Le Ne . FE : | :

Bei grony. ar. Lotmtyldimethylesllyloxy or. an } :

SE J aralkaxy group ag. Lripheoyimeiho:y : Buch )

PIR . tH P Pos ’ : a groupe mar be converted for example by . o

Co h#gipo lve in to the des)red hydroxy gronp oF, by :

SCRE 20 vrancen tert fea on, he conver ted to an a Co Jn . i x Loony T.

Sa fr . alternative apter group. of 3 -azido-3"- Ny

Cap dk ' . eg ’ : cred , . feet ‘

Ce diy thymi ine,

Cn Re - ’ ed Vo i

Bs ‘ CE !

LL : aE en HE .

Cl L . . . Me. Co

Cn REA 1 : : oo i : ~ BAD ORIGINAL 0)

Bi Mi ,

Ter thot

AEE Ln CT : : CF s Se ERG

Chiba oo Te : BAT. Se EES

Rb CL ST 0 sr Cem SEN

AEE a radi Lit : : EI . , . BR

A re fT WE : ee Llgqy LE fd a CH Y Sm

SE WE aE Cl Co COTRMA

Hus ie Se Ta Sipe OL Ie

SERRE : EET ’ . go

HORE | Ee Co

Cet LEE : Lo CT

CE : w : TART ey

Ci wr Ls Cc

LL i “With regard Lo process (CY), this may ba a effeotad for example toy treating the : + : appropriate pyrimidine of formala (IV) or a . £ . : pen . v ok salt or protected derivativa theraot, with a

Sh Bb compound of formula E

Ce i ;

A } : . igh ra 3 oe vo Be ep arena mean SN Sh . + . pat Ee ~ Coe : ot . VL - : , EE :

Ss SEG 3 SRE ib wor Sor Co iat et

CR SE B BN

Cf Ll eT (wherein A represents a leaving group. .8. -an eRe . Lend Lo +

Sal CE IE od

Cp orn agstoxy or bensoyloxy or halo, eg chloro group el

Cnn So as . 1 Co

Caiheseg 0 Red, and: B represents sn optionally protected - Th

RE ey i - ©

Cl ge fe : 1¢ hydroxy group eg a p-toluenesulphonyloxy :

Cl SEE EA : i sroup), and subsequently removing any a an B

Cop a CL

Es \ protecting groups. a co HE . . TY 150 Sy

Ce X: Regarding process (ny, BR) way represent. a } tL Lo . \ > . . . a . - ho precursor group 2a describad above for R. in

NE | ed :

CoE ar ’ = Co Hi .

BAD ORIGINAL 0)

CF cats he ) is LEwia ’ Lo - ab ho to To

GRAPE a Tg QIERE uh iy i CULAR Ee HE Bulh. cn Sg pail aad bi...

Fercitelnder, CY : Cast SL cot t JE CARN gE phd we TR LT Cr SAT agg hep

TH gs RE se = ! Ji “AY : LW ' [a ARSE . ; ot ot 2 ART oo ile cas EY

Tule oo oy Len Sods ride Foo

Wengen To co So Co Te lal Lo CE By

SE AL : Lede . es

Tram se Co Co

CL formnlae (117). 3° -Anldn-2"-deoxvthymidine may

I» \ then, for example, be obtidned Ly reaction with

GT \ fi, oo

Co \ Aan alkall wetal sxida. o.g. lithium agide, ik advantageously in an appropriste solvent such :

RK a , . | . i ~ a 5\ h ae wort MF followed Ly anid or haga » \, hydrolyveis advantage-uizly under mild ho : condi tiong :

RE NG : 3A AGA cdeoxvibymiding ma v he converted oC

Co ) inte a rharmacaulically acoptable phaephate or a © 19 «lber aster tv renction with respectively a :

Ln © phosphorylating agent. e.g. PCIs or © an

ELE appropriate eatarifving sagen. a.g. an-.hoid

RE : halide or mahydride. The compound of forkula dg

Fiera Bo : i wy EE 0 Including enters thereof, may ba cohverbed Co

SI LE 16 Ante eharmaceutically acceptable salte tlhareof Ib

Sg RN apd A ii . . EY

Pena ’ ie : LE : 4

CEI) 1h200nventions) mannar, o.g. by hreatwant: with Ww at Co CI Cnt ¥)

SN AR APPTOpriste bane. An enter or salt of.8°- .

LR agldo-3"-dmoxy thymidine may ba convarted = into

FIAT the parent compound, w.g, by hydrolysis. ©

Te 280 The tolloulng Keamples sre intended for ° : i. ui illustration only and are not intended to limit

SO Ch } ~ a.

Peer Lk Sng . So

Chl Ry CE. :

CoRR cu my : a

SEE TH CL oo .

CA la - 36 :

CEE Eg

Logo Tog RE CL . co cel . Sy SER £0 . So .

RAE Nd SIE LT : pine fins CUE Neg : r RE =»

LT - Ce BAD ORIGINAL“

Sots yi Ce } Cen ° ;

CR So tT _

be RW ie 5 Ss SAGE Eh pha i Fs CUA DE sen AE a he Coe EAE ee es siranasabid io, oo

Sen ERY Lh bE TEE FA Le Ee todd AE SANE Ee go

Ta Ce Co UNS em bo ERY Te eT - AEE Wy TRE AEE

Re Ee #P V bare S eg TNE hy oO . .n FTA Mn \ EL es WL Ee

EIR, Tee Migs So fr sy LOST re Eu

RE A Lan So RANG bsp ip Fo soupy ln Arn Cn . abe LT

Spen oa Ag Ly

Ee . Cy Teen

SE te oo ow

SEER CE Cn :

I . a Cf - the ecops of the invention in any way. The , 1

Lerm ‘active ingvedient an used tn the is. Examplas masns a compornd of formals (1) or a rharmaceutinally accoaptabie derivative thereof. rE 5 Bxamyle 1: Tablat Form lations 4 ~The following formmdstiene SN wp) nN wera prepared hy wget granmiation of the {npradiente

NN with » eolntion of povidone, falleoaed by - a . a addition of puenmeiom ctanrate rnd eomprassion. . . + 18 Formulslion A LL

CO Col : ng/tablet wpeg/tahlat nL oo (a) Aclive ingradient ohne wha Co

Cae (b} lLactope HUF. se ne

Ga Cer . .

Ce SE {q). Povidone BFP. 1h 0 Co

Senta 16 (d). 8adium Starch 20 12 7 a ‘ ol a 3 . Pn ' . Sh | or ' i

Le oo Le, Ulyeollnte Ch

EL So (8) Magnesium Stearste b a

Ce SE ey £6) 200 So

Shi 3 he Lo ¢

GenE oo Co

Co he Ln pr =

LE TRL Le rh hehe NY "s . }

EA SR Nes

SRE . ¥ ERE : SL

LT : oo it Te : id - I = pe So EL ~ on . pr NH )

CE a BAD ORIGINAL OW +

RRS vA Twa re - hl i Com ‘- ro

EEA mT I. “

Hd on id a Le Pie =a’ Nagi ew : : PE ToS I : hE pe Es CUTERGRE ENT dBRL ae cE I AEE A i

DIR Aad LEE : Lo SPE WRI dA UR Get ENT

TART Ln oh oe Cede “Beas bl ae oo oat ad i - Let : Bae

GEE A " Coat

SE AT . Se I nL

EE Formulation B

Sn mi /tablat mg/tablet oo Ce (a) Active ingredient 260 260

Co ! ~~ (b) lactose | 160 - . < 5° (a) Avicel PH 181 60 24 . 2 (d) Povidona B.P. 16 9

Co (&) Sodiom Starab a6 12 wn Alvcollste (f) Mzgnesinm Stearate 6 3 _ : , — a — em. ba 50R wa . . - “5 iS : / .

EL Formulation OC. oo “hl - u mg/ tablet . hy

Ci Adtive ingredient 180 “

Thin Cd RE : CL :

Co : Ligtona : 209 ~ . . " ’ Ja

Ce 16 = Btarch 59

She Eh : Co

Lo . Pov ldone 5 Ta

SIA Po RS Lo

Roe hen . Magnesiuw stearate 4 Sn :

ST : EI A ee an a ve

ER Fe EREE Cd he ) © %The following formulations, D and F, . were

CT 29 prepared by dirsot oompresilon of the rdmixed Go

Cm .. : so lngiedisnte. The lachors used in formalation F RR wag. of the direct decompression type (Dairy oo . Créet - “heparvox”). Co a Be oo FH : - 32 - HE

CE Nl re Lo

Celi Es wr ; J or Ee Wo ER N

CER Sl : pAD ORIGINAL lo

LO SL RT : SE b Apt wf i - > Les - ey . " . . “. ne bed a. . La

Bama. ren cule Ye RERIER Te frp TT Dy EE et RR EN 5d Be A OA ce Tih ve “ 3 po ak ! - : . Tor Sng Ae 3 - ANE Sl pr ’ oi

AEE Lf Re o : oe TING, pu Fe pags JRE RE TRA Lag Lo

Ri rors x L : : SE P \ 2 dy . : yen TO - We oo Ch

LT Wy wt Formulation DB. i mg ‘eapau la

Active Tugrediand, whe

Co Pregelat bypaed pa) C15»

Cc bh HF 10 ee

Co .. 403)

Co Formulation

ToL - ms ean le

Doe Active Ingredient wR

E Lhaghoaa thw)

Tenner AT AY J an A

AC vl) 144A an ee ey -

AL Ki < we

I Formulaotiom F (Oontvafled Weloass Forpalabion) Co i. . Co

TL ’ ~The Lovmmiiabion was prepara ty wet . Bs J E : . . granulation of Lhe inprediante thalonu) aith a co golution of povidone faolloged hy the atitition .

Lo 16 of magnesiom atenrate sod conpresalon.

LAO tal : : - eG i vo uss tabla ; cul a : : Pa) Active dugroedient Ful . a . “ (HY Hydyvorvpropylimathy lee l balan. tie A vo

Ce co (Hathoce) K4H Premiom Co wt = 28 Sle Lmohose BUF. ny : hee oo a ld) Povidons BP. ry

EE Fe ~ {&) Hagnasinm Stearsato 7 i. eo TE rn ion

Si Loon as | :

Claw - 33 : :

So ER . 3 : i . . Teun : | : PA - '

EE Cd TR : oo - ; . x2 j Te oo rY i D -. “ : : BA | INA . } , J gAD ORIG

A So isis part Cy ex oe) $s. ne > a LE att ni Ve , - a :. . nL Lo a 3 Sn, Co Dale Ree big, 3 hi he ay SLUR ET aE. co HEE Tt a 2 gp I PE Amd Li : CLT PHT STOP a a pe LS

San ; CET : Ea - Co Lee %bgqd CT gate RE } Co FE ” PR = , : Lo gy Wel . “ ¥ oS 5 bil ol o

FL A He ws de eee Co EE TL TT yaE a . Sn Ct iy rug release took place aver a period of about, 8-8 honre and wae complete after 12

So hours, : vS ’ - Sn : . i . \ . po | ! ol Example 2: fCspesule Kormulationy

Co 6 Formulation A : - ' A capsule formitation wan preyarved by

Co admixing the fngredispte a7 Forpmlation Din

NG 2xample 1 abhave apd filling into oo bwo-part oo : havd colatine cappode. Faamalation 8 (lofra) .

Co - 19 wag ‘prepared in a similar manner, oo ;

Loo ‘Ferpulaticn = Ll ily bh . cE - SI . MZ Capt he

Co (4) Active ingradient A oe .

FE Cory v . i sri . » {B) hmclase BOP tis . rn Lod 16 (a) Sodiom Shavel Glyeollata oy :

So oo (1) Magnes iam Stearate no .,

LE ) — Te :

Coe : mE " Co : ae REY 40 . A,

Ee > . ~

CL rn CEA : “

Ee Ce Lo Lo

SEE Lo “7 oo

EO ~ Pork al i : ; :

Ll Lo - 34 - sha iy TE .

GU LE sR Co fr Cen ht : CY

Cl Se . ~ An “ . oe at? } .

arin ga oo EE eT laen hee wy So Hest Char eet ey

Bae CO . Ee Doggd

Spl En | Se SOIT

AE at 5 7 EE

SE on oor nn be Co nl Lo

Co 1 : )

FL Formulation C - mig /capsule

Co (a) Active ingredient 7269

Co | (b) Macrogol 4000 BP 66 : 6 a | a

Co ol . : . o. Capsules were prepared by melting - the i» : ; : 3

Si Co Hacrogol Ad% RP, dispersing tha active

CLs ingredient in the melt and filling tha melt

EET Lo : : vy

CES ee . into a two-part hard gelatin aapsule. :

Tras RA "a

So - 7 18, Hormulation D

Td : oo AA .

Fi 0 ¢ B RE Mg/rapsule : a SEE SL i POT .

Got Active ingredient 250)

SA : et . i. So a . dT Leaithin 100 : ino Arachis oil 100 in g a No 16 a } ANG oo . LER Ta Ea aii : J

SE ig re Ce i" } 3 £ . } . .

GET ' Car Cr

Selina £.Capsules were prepared by dispersing the si J motive ingredient in the lecithin and arachis

CETTE Ee : ;

Shas OL oil and filling the dispersion inte eoft, oo SE , Cd Co ’ -.

Cl elastic gelatine capsules. .

Fon hy Co A

SARE ra Co HT Lo

Sree ML SR Co ud oo

LE Ce SR ro A

LE Es SL : wo :

ETE IE v h, ~ fn BOAR i, ob EEE TL a ale 7 Rpt RRR Ld ih a

BR Bs dito tie 0 ee tt wif Fi RT PE Same «TFET A Nr Ete ©

BERRY Be er BL Le UR Ry JA Sh Com MY TNT TR Ls THON 6 Hed SRR go mR aT ER Re

ROIS Eas nA ’ ¥ Vt Saban Te ty ERE)

Fe ER “id et 1 He . . . ’ wd IE n : Le : VN Y g fx oe i

BeotoRLEC CL CEL Ls SEER TR kK: A dat Co ‘ ! : ym : a : } Ay TR % : eer oy Sa : : , Loo

Formulation E_(Contralled Release. Capsule)

Th ~The following controlled relesss oapsule

Ha formulation wad prepared hy extruding ingredients a. b and 0 nsing an axtruder, followed by mpharonisation of the extrudate and oT

Lo drying. The dried pellets were then coated

Ce with release-contrnlling membrane (d) : and

RN NS filled into a two-piece, hard gelatin capsule.

Co ] . mg/capsule

Cn 19 (8) Active Ingredient 260) an (kb): Hicroorystalline Celinlose 126 ad Co (a) Lactose BP 126 oE he oo . NT , Co : ! ann Co (d) Ethyl Cellulose C13 Ll .

WEE ci mmm tel i LSE Low 16 Exampls 3: Iniactatle Formulation :

I a . as God .

WEL ew Ln oY SE : nn

Gemma TL Formulation A. Co

EH . Ln Co

EE { Active ingredient 2.200 g

EAT x “Hydrochloric acid Lo Co

LY =” solution @.1M ~ 9.8. to pH 4.90 to 7.8 ;

CRT So Sodium hydroxide Vo ¢ : wl _-8olution, @.1H © q.8, to pH 4.0 t0°7.9 ” of Con CE .

Sl 20 Starile water 9.8. to 19 m1 oo cob Hn, Cg DE . Si A Car ’ 5 he 3) Co

CH L Vig ’ - 38 ~ BAD ORIGINAL oJ

CM 2 . : Ce LY . nl Tg SL ——-

SEE aos TE . ait Py a Kh si, f Fi

LH h - Joy

Ll CR le oy Los . Co v

I ing EEN

Fa ia gn | Ce CTRL

BES Ras, : ; 5 Lae we IN te 7 3 . ll al ER a ps ; . Rr

WEEE Lo PAE . EE

Fah SE -

Ll a. The active Ingredient wag dissolved {n most oo Of the water (36° _gg0, "MA the oH adiusted to, i : | between 4.9 ang 7.5 WIth the hydvoohiorte aos

OF the sodium hydroxide ag APPropriste. The \ 5 "batoh wae then made up to volume with the water . \

Co - and fi)terad through a ster lan micropore filter : - into » sterile 10mi amber flame vin) (type 1) . \_- and sealed wit 8terile closures and overesals.

ER Formulation p,

Ll 10 Active Ingredient @.128 g

Lo . Bterile, PYrogen- free, pn 7 Le

Le “Phosphate buffer. q. np to 26 m1 : A LC A :

RENEE | Bamele 4: Intramuscurar Indection

Cg Aotive Ingredient 3.20 g

Ce Bensyl Aloohor 010g

EE 16 Glycoturel v5 1.45 g° ho Water and Injection 9.8. to 2.00 1° my ‘ RE =

Sd Lo The active ingredient wag dissolved in ype :

Chesno €lycofurol. ' The benayl alcohol wap then added

VIELE ands. dissolved, and water added to 3 ml. Phe

CEE 5 :

Doane mL 20 mixture wag then filtered through 4 cterile o EE Su CT Co fi pf en oo Lo I

CA iA ,

SE ns i ’ I. } }

ETRE, - - 87 - a

Br Cy RR ., a co

De, Ee Ed I 0 ORIGINAL il

Sasi : : Vo - aay a . ‘ 2 wn 3 : CL

Ta on © se ggg wah : ye

C nioropors filter and sealed in aterile 3 ml

CL : amber glama vials (types 1). : = oo ‘Bxample 6: Ingradientn :

Co | Active Ingradient @. 2600 & oT 5 forbitol Solution 1.6000 g : LC dlycerol 2.9000 &

TE i Sodium Bensmoate 7.9058 g ena : Flavour, Peach 17.42.3169 @.0126 m1 | : a - Purifisd Water q.4. to - 5.0000 ul

Co 18% The active ingredient was dissolved in a : oo - mixture of the glycerol and most of the ’

Ci purified water, An aqueous solution of the :

Ea o ‘ modium benzoate wae then added to the solution, n Cl at _ followsd by addition of the sorbitol solution y

A La 16 . and finally the flavour. The volume was made 1 go 0, LL Co Co he up with purified water and mixed well. . Co

Ph EL A oo LT

ATL TT Rey el tor Cn :

CL Tr 4 a : Co .

SE TRE

Por | - 34 - .

Se en “, 5 I

Sh en ri

CR Ee . BAD ORIGINAL © . ) Hig «0 - . . . .,

b

SAREE BIhv anf 10 200 LC . iS bagless ooo kT . : " a NRA GTO, Lg wi lE au Sl \ Ri: ale FT al SWRA rE Td, Ee VC Le Yo Rh So & 5

BR Tg eR

MIRTRn. ue WR Sw eal SL gg gE bi dE Li I : Ged NEE pr Le AR CER Ee PAE 4 £23 AA a ER to Phat Eg 2 CN “E : Cig ewe eT al CoE Rn

RAL i wT pir

Foner oo PEA Co 5

SN y ce Ce Co

Lo Example 8: Suppository - gy ma /suppository

Active Ingrodisut (63 umit 262 :

Hard Far, RP (dWitepeanl JI16 - 6 - Dynamit HoBel) : 1710 i. oo 2026

TA The activa Juvredient waa used am a o povdnr wherein at lengt 99% oft the particles ‘ ware of R3 nm dlsster or Less.

SEE . C One- FUELED of Lhe Wilapaol W156 yas melted in

Si : 18 aidbesm-inckated pan at 4R°C maximom. The ol Se ani . “ly £0 . } Maa

Ci - afgsive ingredient was gifted throogh a 200 - um

Ce aleve aud added to the molten base wlth mixing, So

CERT vging »'silverson fitted with a cutting. head, le gata CN Sg Co SR

TERRY hy uatil a eancoth diepersion wea achieved. =

Ee go : oy fant eS g 16 MAlntaining (he mixture at A670, the remaining

TERA PRR] . B Sn . . .

LEAS Tie Co com SE . : wHiterssl HIT wae addoad to Lhe vuzpension and os cat ne \ RE i Sn i hE Cg :

PIE ‘ lire] te eanure n bowogeoous edx. The entlre hE - Bugpe neon wine passed through a ztdlum etainless

Pore Es red a . }

Ph ae a TE

Gen gtewl ecreen and, with coubluacas stirring, was

Eyre SE ae

Sima ol 20 allowed te cool te 49°C. At» tempersture’ of

PRE EEE

CEA 3 5 . oA ~ 380 to 42? 2.428 of tha mixture / wae

Che . A oo

LE CTE a

CTL : - Lo nL nt Me Eg - 38 - Ty oo

Po TR CoE So

I RRS LE et

Lola THe Sug . :

Cole CE RE r

PT Fi Jie BAD ORIGINAL 0,

Ponape Ce Rn oe

Lor - TE cf

- . ~q9 a - filled inte suitable plastic moulda. The . [a Vr wl suppositories were allowed to cool at “boom hE

Ee Sh teperature. . . ot

SAR Example 7: Eessaries | oo

Ln EE 6 . I oh Co : Aative Ingradient 83 um 260

Co | Anhydrate Dextroee sg

Ca No : Potato Btarch 383 aT | Magnes Lum Btearate | To

Se | : ma above ingredients wers wixed directly oo : and ‘prssaries preparad by direct compression of

Ge : the, reaulting mixture. Co wi TA Baample 8 ju 3--Aatde-2" =deoxyzt:@= ; - To a solution of 3° -asido-3" ~deoxy ok © thymidine in pyridine (°C), octanoyl chloride ne - Ea (1,2 equivalents) was added. The reaction was : cn allowed to warm to room temperature. When tla ern Ue ER oo | - ee - - 40

Ci EL Ne BAD ORIGINAL 9

NE Ea ,

She Ta Ty

£Cqqy (CHOY 5 HeOR: 2831, cn midica aol) Ind jeoatead aampleta peact tear, the gotnt don yap poarad onto

Joa water. The aguecng phase wan danantad

The vesnlting nl) gar chromatographed on ail ioa t gel with CHOLL Hel, The tilla componnd wan ahtalned as an ail hy evaporation of the polvent from the appropriate fractions,

COHN: cal. (-H4 OR H-F 09 N-17 RD faut, C-F4 AT 1-6 08 N-17.686 1 d7 a6ca, in Tg gota, 6H), DO 6.0306 10M, Ta4.6.4 200.20. 30 And 6 OHNE 4.9-3 sim dH. 4m, O02 3.2. 1m A020 and (CH, of aetanovl)). 0

L.BICA. BH Ig ool Oa BH), © 1.6.0 Aim. 130.5 oatanoyl LOH, (CHG)

Example 9: hi-Acety)-3 -azldo-3 a deoxvthynidine

To a golution of 3 -azldo deoxythvmidine (298) in pyridine {Html at amhlant tamparatape, acetyl ahlorvide (2.1 agoivalents) wag a. lded Tha reaction una ativred top tue 41 : Cw

AL 9

BAD ORIGIN

. Ca

20a henve and kept abt 3 ta & 0 for 20 hemre I+ was ponrad onko bon yater gllh eb irring. The aquaong phaaa wan decanted, The oily product ua diasolved In watar and extracted gfth gatar

TTR TB times), 0.5 HN hvdrochloric acid, water ~~ (2%). snd dv led over masgnagiom anlrhat. The solution ase filleved and avasporated in vacuo

The residual oil wap diaeclved tn chlorafarm, applied ba a asidlica gel oolnmn, aad Flash to chromatographed ve tng ny mest hope |, in ohloraoform Fracstiong with proche Way avaparated and the «ofl was chromatographed agaln using ekhyl aceltsalte: heysna (6:4 v/v

Fractions with pradact ware evaporated in yaoua ta oa whites paella ‘ m.p. 96-9289 anl. (468.6 H-4. 89 N-22 8% fad. (0-468 67 H-4.94 N-22.59

Example 14 : "The following compounds ers rraeparad aacordiug to the procedurs of Examplas 8 or 9 as appropriate from the appropriate sold halide or - 49 r

BAD ORIGINAL - (

3° —Anido-6 ~B3-benrorl 37 -dacxythymidine cal. C-62 BR H-4.77 N18 41 fru. N-63.81 H-4.72 N-18.46 m.p. 64-5070 6 3 -Aztde-- 3 ~deoxp-5 ~@-plevalovithrmidine cal, 0-61.27 H-6.03 N-19.9) fnd. C-H1.07 H-6.,056 N- 19,82 nm. op. 99-1870 3° -Ant1d0-37 -daoxy 5H ~-F-{3- 1& methyThntyvry thymidine pal. C-5@. 24 H-6.13 H-19.62

Fad O-R D7 H-K. 11 H-19.49 : ST. 480A IH Tg oo 1. 2H% BH), 8.13%, IH, 1°HY, of 4.6H-4 16(m. 30, AH and 6 ouzy. &3.8- 4.1F(m, 1H. 4 0), dn a-1.780m, AH,2°H And 5 mathine), : rr

OL.88(43.30,0g gv. 203, bCHZ) da. 00a. 60, 1-6 4Hz, methyle on 5 butyryl) - 43 . )

BAD ORIGINAL 9

E fe -19- BLAS

A A. BE <q fo J» BE” 3'-Azldo-3'-deoxy-5'-Q-palmitoylthymidine cal. C-61.67 H-8.57 N-13.85

SEES find. C-61.85 H-8.59 N-15.7"

Hf Al . i 4

Co 67.65(d,1H,Jg (=1.0Hz,6H), §6.12(t,1H,1'H), 64.5-4.05(m,3H,3'H and 5'CH,), 84.0-3.8(m,1H,4'H)

RY 62.35-2.1 1(m,4H,2'H and (CH,)1 of palmitoyl), §1.8 d, 3H, 35 ,=1.0Hz,5CHy) §1.35-0.6(m,29H,5'palmitoyl(CH,) | sCH3) 3'-Azlido-3'-deaxy-5'-0-toluylthymidine Cal. C-56.10; H-4.97; N-18.17 nd. C-55.88; H-5.00; N-18.09 m.p. 73°C /0 NMR taken in DMsO-d,

NMR: $7.95-7.29 (m, 5H; bH,cH,6H), 86.16 (t,1H,1'H), 84.6-4.4 (m, 3H, 3H, 5H), §6.2-4.0 (m,1H,4'H), 52.39 (s,3H,dCHy), 81.63 (3,3H,5CH;) 31_Azido-3'-deoxythymidine 5'-O-(hydrogen succinate) Cal. C-44.98; H-4.83; N-17.96

Fnd. C-44.90: H-4.77; N-17.85 7€ NMR taken in DMSO-d,

NMR: 57.46 (s,1H,6H), 86.13 (m,1H,1'H), 64.48-4.40 (m,1H,3'H), §4.34-4.20 (m,2H,5'H), §3.99-3.94 {m,1H,4'H), §1.78 (3,3H,5CHS5) . 3'_Azido-3'-deoxy-5'-mesylthymidine Cal. C-38.25; H-4.37; N-20.28; 5-9.28

Fnd. C-38.15; H-4.38; N-20.19:5-9.30 m.a. 253°C (dec)

NMR taken in DMSO-d

NMR: 67.45 (3,34, J¢ g = 1.0 Hz, 6H), §4.15 (£1,354 0 = 6.6 Hz, 1"), y

To §4.54-4.41 (m,3H;3'H,5'H), §4.14-4.02 (m,1H,4'H), 63.24 (s,3H,5'-mesyl CHy),

Ke ~ 51.79 (d; 3H,Jg ( = 1-0 Hz; 5CH,} - ’ “ < LL 31_Azido-5'-0-(3-chlorobenzayl)-3'-deoxy thymidine Cal. C-50.31; H-3.97; N-17.26; C1-8.74 : Fnd. C-50.16: H-4.1"3; N-17.13; C1-8.66 ; NMR taken in DMSO-d fk £11.37 (s,1H,3-NH), §7.98-7.43 (1..,5H;5'-phenyl,6H), 86.17 («+ 1H;

Jy ge 7 03 Fs Ipap = 72H 1H), 84.658-4.48 (rm, 34TH 0,

Jo 84.14-%.11 (m,1H,a'H), §2.48-2.41 (m,2H,2% 1), §1.64 (d,3H4 3g ¢ = 1240 5CH-)

HDL /I.MJ/26th February 1986 . ea — rte io BAD ORIGINAL = a ig cr . 5 . oo Batman ] | ) p Sath — — . = fl Pe - ) : NS 2 Ee _— " A Stara esse. €. a wo Cg tae ~~ - 15 il be

PERE a COTTREIY Dh EEA FU ER EE

SSE Es oC Co COR “lssihall tn fA FF * ee ET apr IR Vo bess

Eo oo 2 : WE : Example AL. 2.6 =0-Anhvdro-13"~asidao- a denxvthymidine

CL | 3’-Apido-3" -deoxythymidine (3.9%. 11.2 mMol) was mesyleted by the addition of 6 mwathaneesulphonyl ohloride (2.7 ml) to a - solution of the starting material In dry

CL “pyridine (2 ml). The reaction was allowed to

CS progeed at 5°C for one hour, then poured onto

Co ice water. The precipitate was collected by oe

He 19 filtration. The product (3°-azido-B’- ll Lo mesylthymidine were heated in an 82°C oll bath

Ly SUNLEENS forsix hours, then pourad intd ioe water. The

SET product wae extracted from the water with ethyl

LE : soatate. The solvent was removed in vagua and

SI 16 the resultant oil was falsh chromatographed on

CTY Co silica gol by slution with . CHO14:MHeON (911

Cn ; Co

SE v/v). The title compound was obtained am a iE - " solid after evaporation of the solvent, trom the nih leer appropriate fractions. dF 28 wap. = 184 - 186°C | Go

Fe ; “ : i: vn vo co Ch : T .

SE | ” 0 Cte,

ST Se Co - 45 - BAD ORIGINAL J Ct

RAE nity, 0 Ee Cn ee ACNE CET en Sn Th Ah nt Mail Ll nem ce ts

Bf nn oT Re sie

Re Cbs LE oo Co CRT +, SOR Race

TE $8 vy, Te Cer 2 Co t = & ay % i aly Es qu0 Ey 3 Ed Pr Jog ov : - LE i ' . . Lo % Ba) 5 tars :

ERE a SN Cn URS Bh wk Ln CL oR

EL Ch Se en

J Ko, : co : ‘

RET : :

Toad b sl . :

Cv

Wo . . . £ Example 12: 00 Aaldo. 3 cdotss Whowld Les r

AE , he : : ;

Ed (ue) 2.00 CAnhydrethymiding

Cha

SR Thymidine (Bh 4 g: 9. 30D wol) wae Adamo lvead

WH _ fr. he - oo : ) Joao BEX wm) dry DHE and added bo Nha i Co Le > 1. 1 ,2 Le EfIncrasthy Lidl ethy Taming E148) &

SEE do . 2.0628 wold (gprapnrad according tor they method oF a D.&. Avar., FV. Hel Chow fi, 608 ( 196) Thi } fe bore ’ fy po

ST aolutlon wag haslod nt TOC for 9 misutoa thew dT poured Like B63 sl pthonrol (KLOH) with vigorons oo Eo 2 12 stivy lug. Tha produst graclintstod (rom thle

Cla solution and wap £1) brad, The EEOW

Coben Va vo

Cdr RAE

HS suparaalant va yvofrigoratasd thon fittorad Lo

LoL an oe }

The oC 181d thes biLle compound. mp. -228 23@°, 1 "

Cyr Ea : Le

Codey LRT , I

ST | CR a ty ik a oA) 3. -Aaide: 3 adouxyihyriding a So

Col vy 16 “2,37 -@-Aohydrathymld ine ( nh gr 9.1116 wold

SE 0 : and“ Hally (29g 0.446 wal was ewsponded In : 3 aulxtinre of 260 mt DHF and 38 ml water, Fhe t a : By R i . : ! i

So rabetion mixture waa refluxed Cor & hours “ab :

PI which time 11 wan pourad Jute 1 Titer of wator, j

LT a Lo j

Cu C20 The aguacus goluflon wes exlracted with ‘BOOAC er CE : oo ' " : (3x 2% ml), ‘The ELOAC cutracts wera deled ae s oT v. SE . tf ek TL i | aA

Lei : Er D Gren x Fe . : . © i Lo \ _ : - 7 . . :

RELL alee oo CT REE ERR

Then ES | Cad Gee ame oR | | EE Hl ha a | i

Cee over Na,80,, Filterad and the BLOAc waa removed - oo — ee An vacuo to yleld a viscous oll. This oll wae

ST . stirred with 2090 ml water providing the title

Ee . compound. ap a eolid whlch wes anllected hy

LY 6 filtration. wp = 116-118°C J - Exauple 13: Hoposodium Salt of 3 -Aaxide=dl-

Co deoxythymidine :

Co (_.

LEN © Ly. Approximately la of 8 -azido-3 -

Can St - deoxy thymidine ans dissolved In BP wl. © of

FT | 19 distilled water. Ths pH was adjusted to 12

SE with IN NaN. Approximately half of 5 the

Co SE solution was freeze-dried. The title compound or was obtained as a lyophilised powder. vo «

EL Analyeis calculated for CyaH1NsiA046/10 NpO ed . - CAEL AE 3 v Foi a. 16 cali C-40.93; H-4.47; N-23.24; Ha-7.66 1 ;

Sb tnd: C-39.88; H-4.34; N-23.23; Mn-7.99 Co

AA . Example. 14: Freparakion of 6° -Honophorrhals of oo

Cope © 3irAaido:3 =deoxyihenidine CE i to 3'-Azido-3 -deoxythymidine (8.5 «, 1.87 mmol)

A wn af j Po

LL fo = i Ba co : " I. Ra }

Sop Si : ER - 47 - Ey :

Pl ST SOE | cei 5 ln, Lr A | BAD ORIGINAL §)

A . : Co , .

JOINT : . + : wh t _ . : 1 t" wan dissolved in 6 ml of triethyl phosphate and §

EE the "mixture was cooled to -5°C. Phosphate oxyghloride (#.686 ml, 7 wmol) was added in one portion to the rapidly etirred solution which :

Co fa . i. g Co 5 was then maintained at -1970 for 22 hours, An } Pp ? Ce TY, . co

Cy aliguot was removed and added to concentrated : | oh ammonium hydroxide. Analysis of thie samples on i

Lo TLO {eellnlose, n-ProH:Hy0, 713 v/v) showed no

Lo Z remaining starting material and a eingle .

FT } 18 fldorescent spot with lower mobility than the | :

Rei - Si nuoleoatds. The reschlon mixture was poured to

RS ‘ : So Ee : Co AR i.

RI . onto’ 20 ml of 1lce and water. this was placed Nn

Srl PR I. oo

Ee A Sin oan ice bath and the pH of the soluiion. wae Me

Ch A © adjusted to a value to a value of 7.6 by the ; he Vo a Co : of : - 4

Er 15 addition of 2ZN NaOH. The basic mixture Was wd - Co extracted once with chloroform and once with

Si ai : gther. The aqueous layer was again adjueted to Co

TAT Cort Sh SA J C Lo

SE so alve. a pH of 7.5 and concentrated in _yaquo to :

Gren bo remove residual organic solvent. The material ; : + , = . ; . SL . ie J

BE Li 20 was stored at -19°C until purified as follows: :

Tel CE Co

Co att a Co DE

SR CE Deactivated charcoal was prepared by washing : :

FEE Re de Cel cE i

SESE aoddnut charcoal (bO-200 wesh, 120g) with ' 500 Se

Hk Vie TW . -

Are Cy - Con

Loh xs 3

Lal igh a BAD ORIGINAL .

Ei PR FES A a 23 Pie Sgt Een a a et a Te Ln ALR isn Tae aA bil

RETO PPS A Cy ji TARR ee Fae O15 hh a Re

SWE RS nT a EE es i a Pe

FEST BL TLE Sie BG HM STE

EIA Th TTY TMT SLR CE Ke oY SE ey REN: Jey : Jared Ca LT ’ CEST : . leah To co EBs Jed Us HE pLLC Le - Co SE Con Red front £2 EST ESR en LE a Chg dh Sd a RRA Gg \ PRESTR ’ . eine VaR Ee [or

ST eA : 4 9. Ca ’ Sl . Site Foe i Bl ef UA ool WER

SEL LL : \ a ’ . Dt en el eo ; | Re

Toys ooo ’ or : !

Sr ml ef Ut NHCL, 3 FL of water, 36 ml of 3% a toluene in YE% ethanol, GOA wl of 668% athanol i and . finally exten lvaly with unter. ! Paactivataed charcoal (12 wl =f aA=ttled wer : b charcoal) wae nddad with sticvicg to the monophosphats wolulbion (B.72 g, 1.3 wold, 39

CL ~ mh). The supernatant wan decaated snd the

CL . oo charcoal wan washed uitl LED wl of yalbeyr. “The:

CNL nucleotide wen elubed bros the elinyeanl by 180 washing with 129 wl) ol 1.01} smweon bog bhydreiite

EE Cla 8 othaved Thu solullon wan f11teved

Thee } through a ©.2%2 meron {filler conan brat ads La ade LT vagy to 18 wm). Filherad Lhroagh oo Awlodn

Can ch Centrlilo OF-26 wembrans. and Tvophtiland to aE 16 yield the Adams bom A azido-3°-

Sen ET . Ca

Cdmt nL daoxyiLhymidins- Bb -nonophosphate a8 4 solid, ite JERS Co CL

Hera GM ’ ;

ARE CE Thig gomponnd was characterized as a nue lsoglda wide ESE 6 ~menophosphale by thes Cnbittis of

Sorat Lee BEE

CRE Do 6 pnubleovidase Lo degrade It to Lhe nua leonide.

LER CL Cl

CLE Ee Ci -

Cote Ce 26 RKxamelo lbs Goosen Dc lrivherrhatseid] © : i wo Yo Jacky bbhymiding ahd:-3 Aside bl dlohon bo bunds :

Sper tr ve dT wl Coe i Si To a 2 - LE : . re co deexythimnidine a.

SRL Eo 3 1, . = :

S5 : JE i . _ nT : eH “i SO :

Fe con wR .

SE a 49 = -

CERN E Co aE rT fo 3. . .

CELE :

Cpr ray | : :

CoE Tir ;

CT RE Ll

Coa ne BAD ORIGINAL

CERT eh Ce Co

Sey pe A [SA . Bop : . 3

Cae

BACH

SERIES CT Ti Te Ee Te a AR ea es RELL

HN Een SL RRR bn ET en adn a Lig iE

SE aR Tee en TT AS RE DEANE

EEE To ELE Loe Coe Thi : v3 EA: 08 © oN Te Ee SR Ce Mae CRE) Ee

PERE Senet : ; Ee LEE aT Ee

SEE eT RL gn CRRA =o SE IN . Lo TR . ToT

Co (a) Bla.(tel-bukylamvonivm)pyrophoesphinta : ‘A column of DOW BB ("Dawex’ isn axchange resin- DOW Chemlasl GLsbhoratories) puridiniom

Co resin was prepared by pouring 49 mt of rosin . fete b into a 26 om dismeber aoluma snd waphed with : i : rater until Ke nore aolanr ented,

Lo Pyroplioaphs be decahvdrata (1.12 ag. 7 v1 mM) wan eee dlasotved Tu 20 wl of water wy! applind ba Lhe

TNC

Lo es luamn The column was alatad with pater, © A :

ST SARE : .

ET 19 128° wml frachion of Lhe alnant uhinh contained hn , : Ch E

Lo UY. shsoxbing wmaterlal was collected The

Coe volumes wan raduasd bo 10 milly vacuosnd tird-n-

Sr Ski 4 . : iB Soe mo : bufylamins (1.2 ml) wae added. Pha vorlima wap roaducadin. vagun and Lhe ragidne wan deiad by

RE 15 aaavaporatiton alth pyridine fan Limen., © The : dle CT product was ntored in a freene (~B°0

Ca NY ii b 5 nT (Lb) Wydrogea form of 37-4aide-b monovhoiphake-

So a=decxy bhymidie Lo :

Cr k The: ‘hydrogen form of the monophosphate was Co

EL Ck Pon Lo for .. 20 prepared by passing the ammonium malt. obtained

EE - . ol © } - >

RT Ca pd, \

Te B A Cem } cn - ha -

CL

SL,

CEL ml - BAD ORIGINAL

Re Lo SE - bes

’

Fo Ee hE P CE ag hae - : - Ca Co no . To Ce ad Rm Bs wl EE eh ce - * : In Bxample 14 (A.1 g. 4.283 mMal) disanlved In

N 8 mh of umtar, Lhrough a 1.6 mh (12 eg ) column

Co of NOW 63 H'. to) Fhosphoromorphalidate Derivative of | 3- oo E._. Anids-3 -deoxythymidine - In 89 ml of water was dimgolved @.283 mMal of the hydrogen form of the monophosphate obtained

Co \_ in. stags b}. HMorpholine (98 ul, 1,13 mMal, 4

B Co. eq.) was added and the solution heated to Co 18 reflux. Dicyslohexyl carbodiimide (0.234. g, i 1.13 mMal, 4 eq.) diseolved in t-butanol (6 ml) 5 ni was, added over a three hour. period. . The 3

EE . Co raaotion wag refluxed overnight. The reaction ~ | wae cooled to room temparature, filtered, and

I 1b the solvents. remaved Jn. _xacug. H®thanol was

NU added and evaporated in. vacuo four timas, The resldue was diesclved 1n methanol and the rhosphoramorphol {date precipl tated hy the ) yoo ‘addition of «ather. Theo precipitate wap

Co 28 trituvated with ether four tiwas snd dried on a : . rotary. evaporator. The title compound wae 1 | obtained. 4 2 \ ar EE

Les BL v - - Bl ~

As = a BAD ORIGINAL Y

Ta . | \

0S se fenlgs se - TOE mo oe

IEE oo phen rr 6 ve sae 2 (d) 1° -Asida-3 -deozythymidine-f -brihesphate tl The phuaphoramerpholidate derivative obtained in atage co), was dried by A removal of pyridine - in__ _vagua four buimes. The bile (n-Bu)gN

Hb pyrophoaphate obtained in stage a) was aleo - dried by removal of pyridine in. _ vYAGDO. Tha

Co ] ; phosphoramorpholidate wae Aineolved in 3 pyridine, F sb, and added to the vessel : — ' containing the pyrophosphate reagent. The bh . 19 reaction wae allowed Lo continne overnight at : ws room temperature. The pyridine une removed 1n

A YAQUQ. Hater was added to ‘the residue and

CL removed in vacuo three times. The residue Was ~ o frozen. . 15 The residue war thawed and dirsolved in 58 nl = of water. The solution was applied to a column

NY (1 x' 10 om) of DFAF Baphadex A-25 which had

CL been equilibrated with = 59 mH ammonium ) o oo blecarhonate. The phosphates were aluted with a

Lo oo 20 300 ml kipear gradient of b@-8aaA mH ammonium : - Co bicarbonate. The fractions containing the oo . diphosphate nucleotide were pcoled as were oe =. | ; Cn -

LL Coe ES BAD ORIGINAL DJ

4 a . gq

Te

Co. RE ; thease containing the treiphnephate mnlantide. oo The pooled diphosphate and triphos hate : a fractions were each dried in vacuo. redissolved oo in water, dried ngain, redimeclved in water and : 6H Lyophilized.

Example 18: Enzymatic Synthesls of 3 -Azlda=p’- : . triphospha ted -deoxvthyeldine . NE : 3 The ©'-triphosphate wag aynthemized from the . 4 . 5‘ -diphorphate nging pyruvate kKinnge and - i 19 nucleoside diphosphate kinase. The reaction

BB mixture contained: 6mM 8 -amida TDP. 12. wi adenas ine triphorphate, AA mH HaCl,, 49 mH : potassium J toarantna-N, Ii ~binl2-athunaan phonic ! aotd) PIPES buffer (pil 8.8), 6 mM

NN 1b phosphenclpyravate, 4 10/m} maleoside ‘diphosphate kinase and 194 Ti/ml pyruvate ; kinase in a final volume of 6 ml. Tha reaction oy © mixture wan incubated at 37°C for 6 daya. The

Cl | ‘reaction mixture was applied to a column {2.5 x ) Ra ~ 20 1 om) of DEAR Sephadex A-Z6 which had been

Si equilibrated with ammoninm hicarbonate, The

RE Lo ; . oo 5 | ~ B3 -

Co B oo . aD ORIGINAL J)

CEReEAT oo ar oo CL ed > fe : ; 3 : h i : . - el | Le EAE : 3 ane oo = siakd . - nucleotides wera alnted with a gradient of {ow ~ 1000 md ammoniom hicarbonate. Fractions containing tha triphosphate were pooled, and

Ce : avéporated to dryneas in _vanuo. The <ompouund 3 b was further purified using » preparative HPLC : i . column (Whatman, Ine., Hagnum 9 8AK)Y eluted }

Lor : with a gradient of 18 - 1 mM potaasiom o | “phosphate, pH 3.6. The resnlting compound wae

Ww. , further purified uweing » DEAE Sephadex A-26

SEE 19 column as above. The fractions containing the pE tetraamhoninm 3" -azido-3" -dsoxythymidine-6"~ he - DE triphosphate were pooled, dried in _ vacua, b Sree memes - FA 1 8B 0 1 VA in water and lyophilised to yield oki the title componnd

Fat : oo oo ’ me : i : 16 Example L7: Antiviral Activity Lo ; ce : (a) (1) Betrovirua - Ioduged Malignanoy

SER - 3" -Anido-3°-deoxythymidine was administered oo

RS to female BALB/c mice Infected with 1.6xint

SE Pfn of the RYB3 strain of Rauscher Murine

Cha oo 20 Leukaemia Virus. Treatment wns nlarbed 4 : i [.:. hours after infection at dosnges of 89 =

CY Co De a bo 3h - BA - wl 5 ‘4 T BAU UesiailvAL 9

Cn " He ah } or } Ce ’ ‘ | vs re -

Geme s .. Ca Ee : 0 Sees ce ERIE to Lo LE

Coa pn Eads . = CTT eee

FET Lie Co le st owl vf 3 tl Ee

RAR © mg/kg intrapariteonaenlly avery 8 hours or . LL a 9.6 or 1.8 mg/ml orally in drinking water.

She Suoh treatmant wap found to prevent ’

Se : : vo oo . infection of apleen calls and subsequent oR iy H . davelopment of eplenowegaly and also

Cb auppressed viraemin. : : (11) HILY-I : : . : Ww. | .

Ch © TM-11 cells (T-cell clone susceptible to oo iE ) | « HTLV@I infection) were co-cultivated with

BT 18 7. irradiated, HTLV-1 producer MJ-tumour cells

TU as follows: dh Ta) TM-11 cells only; oo oo _

Ee 4 : Sa :

AEE oo #2 Db) TH-11 cells end WJ-tumour cells

Cake Co Sl : | -

Sar LE “Xs e) TH-11 cells, MJ-tumour cells and 3° mal Lo Ct Co : :

Th oo 16 “ azido-3 -deoxythymidine (3uM); . BE

EOI Cd) TH-11 cells, MJ-tumour celle and = 3'- he wr azido-3°-deoxythymidine (BuM): oo } : tl , , . .

So a ...e) TH-11 cells, HJ-tumour celle and 3°-

ES EY PE co ood azido-3 -deoxythymidine (27um). : . cd

GoFm CL . EE | “ : pF i ~ \ nL K . : >

Ly Lr . on ] - - BH - 2

LE TL : : ~

SLRs ET hig AE i

CERN - BAD ORIGINAL ¢ ; LE : “ Va } # i i :

E ff a

Ei i ; k. | Dnoday 18, total PHA Wak extracted from esch i : a culture apd dlgeasted with Bam H1 to generate gm 4 Fragment, of the HTLV fonone, independant, of

E any host Flanking sequence and having A i 5 standard wolecnlsre welght. of 3.3 kn. The: ? digest was then probed with radico-labelled 4 Tanbidha [Hr-o vA Shandard probe recognized the 4 Baw HI fragusnt of HTLV : No hybridisation Was ohaarye) for BH), : 143 indicating & tack of virng in the min feched control. A strong Blgnal wee peep for b), the ntreatedn infected central. A yesk slgnal was aheervad with er), Tnaicat ing incomplete : eradication of the virue, snd po hybridisation

Wan noted in od) roe) Indicating com plete

Sateradnat ion of the viru.

Each cid ture Wag alsa probed wil A probes fap

Tell Veceptor B ohain, with x strong signal being generated For a11 Girl bre | showing the 24d contiinoed pregomoe of TH-11 Foy the duration of

Lhe per inen to. : - he ig 8 i a. Fr . i

TA i

Ny P Po ti 3 E 3 p

TH Ek i 3 . bh phi: pi I

A } i . l 3 !

I. i

Bi: ' } a

A ; (by AIDY

PE : +34 K l 3 4 - re (1) Heverse Transcriptase Antlvity

K ; . . or i

G : 3 -Azide-b -triphosphate-3 ~deoxythymidine i -

E | was tested in. yliro against ALDVY : 1 5 transoriptase (ATDY RT). : . 3 V + y ATV RT was purified from pellated and 2 i extracted AIDV by elution through DEAE and 1 phosphonel lulose caolamns., The enzyme . - sotivity was linear through 66) minutes and 12 stable for at least 2 months when stored in fon glvoerol and omg bovine serum albumin ; per mal. Using rA-odT yo 18) ans rhe 1 template primer, ATV RT had a pi opt imnm

Of Tov ta 1.3, a MnCl, optimus © f @.3 wd 1h did on Hatha opt foam of 5 mM. The act iviny in the presesnoe of 6 mt Mal. wan te fold vreater than Lhe motivity in the presence ; SF vod mil Mola Musimal enzymes wsotlv ity ane alas found in Gey to 148 wh KO and tH) “i feo 10 wi Hall, jncorporation of [PH] dTTP

VEE Piiear with respaent [I crf E Ye ] SET i vont nl rat bon, When bepshed, 3 -azido-5 triphosphate-3 deoxythviddine wag Found to bee 5 Competitive inhibitor o fr AIDV RT, ; giving un Ki of @.349 ub when vaing rA-od| 14 15, an the templale-priver. The: wasyme had 4 Km for ATTR oF 4, 81aH, ahiggeating that

A aslde-5 -triphosphate-3 deoxy thymidine binds tigher fo the ansyme than Aven dATTE,

Further expercibsnts with the KT as of avian 1d myeotobhlaataosis CR ERIEN Moloney mil ise bankemdis virus aid ADV, showed 4 azido-

H triphosphate 50 ~decuythyvinidine to Le a terminator of DHA chain elongation, : . J (11) lin. Yitro Anti-AIDV Activity 4 Aldi 3 deny thymidine Was treated and i found to pogseas activity in a nawnber of jpn ’ vitro saa Svs bans, Price af feet FIs Te '

Cl mensurad hy Ansaving rovepae transcriptase . {73 aoldeity dn the superna Lim From

SE inland, Uninet, Ail pay ores rated ctl ls 3 Amide 3 cdeonvihiving die ,

HE

( -

BAD ORIGINAL iv ]

He ; : "i CL Lo affactivaly blocked Lhe Infection hv AIDVof —~ | the HI nnd 1237 hmman lymphoblastoid call 4 liner nt concentrations from 2.7 to a.o313

Ca mor /ml. Similarly, infechion of normal FHA ay b atimylsted white blood cells and cultured hi peripheral blood lympbocyber wan inhibited at drug couvcentratlope as low as 3.413

So mg/ml. Dmg sddition and enbtrachien

Ci WW. experiments in W9 calls revealad that 37- oe 1 pzldn-3" ~deoxythymidine was mont affective

Si when preaent, at, tha Lime of viura co infection of suacephibia cells, bunt still :

Sh oo retained most of itn antiviral ant ivity iy aven whan addad ap lata aes 2A home after a .

Cp 16 initin] AIDV infection. [ohibition of

LE viral replication uns also evident when the

A k drug wee present in the media only during fo ~ the 20 hour period of viruve aneorption. a RLfecta ware aaen At @.13 and $.413 mog/ml.

A 20 3° -Azido-3°-deoxythymidine exbibited no

CE direct anti-RT activity against purified

Se AIDV virions. gimilarly, the drug had i 11ttle or no affect on the production and

Cu | BAD ORIGINAL PN pu i; k - - bas F

A

A

Fi i 3 i

N

; release of wvirvrous from the chronically 4 infected HY AIDV cell line. i ; 1 (111y Froveoting Infection by AIRY } The sbility of 3 -mzide-3 -deoxythymnidine to block infection of cells by AIDV was ! determined as follows. . : ¥ Cloned T4 positive Lets specific T ’ be: lper lymphoovtas were infected with a fro of ATDY lpalates [at challenge doses 14 of up to BEY virions /oell and cell survival after infection was monitored.

After 14d dave 1a culture 10 viral : ovtopathic affects were seen in infected T cells Treated with 8.08 and 1.3 meg/ml 37 - agido- 3d -deosythymidinsg, while antreated, infected cells were H-fold decressed. Cell survival was sleo evaluated in an HTLY-1

Ly ans Format, ATHY snperintected cell linea derived From Lhe cells sbave. A -azido-30 iA deoxythymi tine ab concentrations of 2.7, 2 and A013 aicg/ml totally blockead ~B# .

aytopathet io af feos at 7 dave. Pymtective effants gare geen in Infootionn Induced hy beth cell free virions snd call pepociated virue. 3. Amtede-3 cdecrythvmidine at 3.27

H meg /ml concentration alao af fertively prevenlarl Autopathic ffact indnetion by A laps ralatad Haltian igolate of ATDY. ; Example 18: Tosiolty Arsay

A —antdo-3 dengythymidine uoa admintcterad Lo 18 both mies snd rate. The Lbgg value une in ences of T6500 mg/kg in berth rperien,

Claims (3)

1. oo ZL } ! ? . Claims: ! 69 9 4

   !. A method for the traatment or prophelakie of Aan WTLV-T Anfection in a hnmon which comprisae sdministrating = tharapantinally b affective amount of A cantdo- 3 deaxvthemid ine
2. 2. A mathod for the treatment of an WTLV-] Positive lankaemia or lymphoma in a haiman which - comprinras adminietrating a thavapantically “© etfective amount nf 3 caride 3 -denrvthymidina 19 in a therapeutically effective amonnt
3. 3. A mathod for the treatmant or prophylaxis , | of an HTLV-1 infeaticn In a human which - comprises admintiataring a therapentically affentivae amon of a pharmaceutical ~ 16 formulation containing ar act fue Ingrediant, : 3 -aaldo-3 ~deovvthvmid ina / - 62 - r BAD ORIGINAL 9 L