Synthesis of Ami nosriga r Nucblcos ide%'
Several hlocked glucosamine-pyrimidine iiucleosides were synthesized from l-chloro-3,4,6-tri-0-acetyl-2-deo~~-~-aceta1nido-, 2-caIt~obenzylosynmino-, and 2carhomethoxyamino-o-glucopyranose (I, I1 nnd 111,respect.ivel>-)by the HilhertJohnson and the acetylcytosine-me1 cury procedures. Interconversions of thcsc nucleosides showed that they all had the same configuration at, the riucleosidic center. This was unequivocally established to be 6 hy periodate fission of 1-(2deoxy-2-amino-p-o-glucopyranosylj-cytosine (XIII) to the knon-n dinldehydc XXIII, and in addition by cleavage studies on the uracil XP. Hydrogenolysis of the carbobenzyloxy-nucleoside VI11 gave SXVI, reductive methylation of which afforded the dimethylariiinonucleoside SXVII. XXYII roiild also Iw rnnde by hydrogenolysis of V to XXVIII, reductive methylation of XXVIII t,o the dimethylamino compountl XSIX and mimonol~-sisof SXTX to SIV follo~wcl I J acetylation ~ of XIV.
The antibiotic amicetin, isolated from Sireptonayces p l i c a t i d has heen shown to h a w a structure4 incorporating an aldosyl-cytosinc nucleoside with ai1 amiiiosugar (iii :i disawharide link Tvith thc aldosyl moiety) and a peptide ( 0 1 1 the cytosine moiety) -1micetin and two other similar antibiotics, baniicetin t ~ i i dplicacetiii, isolated from the same fermentatioii,3 exhibit iiiterestiiig ailtibacterial and antitumor activities A comparative study of the activities of t h e v antibiotics and their derivatives5 led to the speculatioil that amino3ugar-pyrimidine iiucleosides with ai 1 unblocked amit io group in t hc yugar residue aiid an acylated amino group on the cytobiiie moiety may possess similar properties. This paper reports the results of a -tudy directed toward the synthesis of bimple amiiiosugar -pyrimidine iiiicleosides that would incorporate these features and yerve ny modelfor the Tynthwis of amicetiii ;\lthough the \viithezis of a varietv of amiiiosiigar-puriiip iiucleoside'i6has heclri stiniulat et1 1 ) -truc>tuI:II ~ 11 I'irsrnteii b t f o i r the l h i i r i o n of Oiganic C l c r r n l . t r \ l i i t l l \ I t c t i n l >t rilt i i i i ~ r i Ii I I ('heinical Society Boston, h I a s nfiguratioii in thiv reaction could bc utilized to get XX. while a iict i r i ~ ~ s i onioi d~d~still be useful to obtaiii :I mannose nucleoside which could not hi. made via the TIilhert-Johnson rcactioii . 1 3 b I-Ton-c\-cr. :dthough hoth XV :*rid XXT7 c \ ~ i l ~ eiiid trogeii oil tretttmciit n ith nitrow arid. uracil imcl a gummy rtduc*iiig Yugar w r c ~the o1ily products irom X\r, XT hile the produvt from XX\o t i tltw~+ylatioiiyieldcd oiily :I guni aiicl not cr
+
+
iot i ()t 1 - ~ ~ - ~ I ~ ~ ~ ~ ~ ~ ~ - ~I I -~ *~ ( iJ ~i )~~I ( K~I-?Li II iJ i I I I spit(' of i i i i m r o u ~rittvrn1)ti, i l i ( x ~ ) k t * : ~ it xv l l I< \ \ n i t t i ' J. 1) J l i i t i i i t i i t r i d 0 3 3 ) 13 \\ Jrarili,~a n d 1. 1 orirliit li! I 1) 1 witnotr ( 2 0 ) I n refercncL [2i, I i 1: I, I~ciixf?tie-et,her-~~etroleiini ether niixt>iirehat1 the s : ~ i i ornrlt hiit after drying for 24 hr. irr uacim at room t~enipc.r:~t~iii~. Ili(1 i i i c ' l t i i i ~ : [)oi1it rosib t(J82-84", wit,h some prior shrinking. rfrtal. Caled. for C14H20C11\;OY: C, 44.01: H, 5 . 2 5 : ('I, !1,2!l. I ~ o u r i ( 1 : ( ' , 14.20, 44.55: H, 5 A i , 5.49; C1, 9.22, 9.24. I-(3,4,6-Tri-0-acetyI-2-deoxy-2-carbomethoxy~mino-~-~-~lucopyr~nosy~ ) -4methoxy-2-pyrimidinoneCVI).-A solution of 0.6 g. OS c~hlorosrigitr111 :mtl 0.6 nil. of 2,4-dimethoxypyrirnidine in 2 ml. of dry benzene was refluxed on thcd steaiii bath. At t.he end of 5 hr., a solid began t o separate. After 20 lir., 20 1111. of ether was added t o the cooled mixture. After filtering and washing with ether, 0.02 g. (84%) of the nucleoside (VI) was obtained, m.p. 206-20XO. Recrystallization from methanol-ether gave slender needles, m.p. 207-2008': ! O ( ) ? ~ U 1 .:1 in CHCla);::;EL 275 mp ( e 5,040). .4,raZ. Calcd. for C~IUH2,SI0,,: C', 4X.41: H, 5.:3.5; S , S.!)l. Found: (I. $8.51; €I, 5.47; K,8.92. ,immonolysis of 0.4 g. of the nucleoside hy i,hc usu:il procedure yielded 0.14 K . (5254) of the urea S I I : m.p. 275-278' dcc.. nndeyressed by ndinistiirc~wit11 t Iic sample frnm the previous preparaticrn. 'Thv :irvtyl (1eriv:itivc. W:IS :11$1i l)repared, m.p. and mixture 1n.p. 268-264' tier. 1-(3,4,6-Tri-O-acetyl-2-deoxy-2-carbomethoxyam~no-p-~-glucopyranosyl~ -4acetamido-2-pyrimidinone (1X).--A4 solntion of 3.82 g . (10 nimoles) of c*hlorosiig:tr I11 in 40 ml. of toluene was added to an itzeotropically dried suspension of 3.51 g. ( 10 mmoles) of :Lcet?-lcytosine-mercLiry in 150 ml. of toluene. Aftrr refluxing with stirring fnr 20 min., :L second lot of B.x% g. (10 mmoles) of the chlorosuger in 40 ml. of toluene was added and the refluxing continued for 80 min. when complete s o l ~ h o noccurred. BJ. employing the usual work-up, a crystalline solid W:LS ~)btained. One recrystallization from alcohol-ether gave 3.28 g. (65(%) of
rrop, 0.10 E., sitit,
+
(4.5) This conigound map perhaps he the oxazolone derivative, 111.1). 174-175° desrrihed IJS Konstas, I. Photaki, a n d L. Zervas, Chrm. B e r . , 92, 1288 (19.59). These authors mailr tlria i,rrinpound by tile action of titanium tetrachloride or a mixture of i J h o s g h u ~ l l Soxsch1oride ~ l i i i 4.
nluminum ?kiloride on ~ . : i . ~ , R - t e t r a - O - a r r t y l - ~ - ~ ~ e o x p - Z - c a ~ b ~ h e n ~ p l o x y a n ~ i ~ ~ ~ ~ - ~ nose and ])ostulated ita formation t h r o n g h t h e chlorosrigar (11) i n t e r m d i a t c .
Sovember, 1962
SYSTHESIS OF
AMIKOSLJG.4R NUCLEOSIDES
1139
nucleoside IX, m.p. 960-261 '. An analytical sample, recrystallized from alcohol, formed slender needles, m.p. 261-262'; [ a ] " D 4.5" (c 0.94 in CHCI,); XE;E$ 249 mp ( e 17,300), 299 mp ( e 6,780). ilnal. Calcd. for C?&&4011: C, 48.20; H, 5.26; 3,11.24. Found: C, 48.04; H, 5.32; N, 11.34. Ammonolysis of the nucleoside IX gave the urea X I , m.p. and mixture n1.1,. 275-278' dec., confirmed further by comparison of the acetyl derivatives. l-(2-Deoxy-2-acetamido-~-~-g~ucopyranosy~ )-uracil (XvI).-A solution of 0.5 g. of nucleoside I V in 7 ml. of warm methanol was added to 3 ml. of ethanol containing 0.8 g. of hydrogen chloride. After 24 hr., the slightly brown solution was evaporated to dryness in vacuo yielding 0.345 g. of a white froth which was dissolved in hot alcohol. On cooling, thick cubes were deposited. These were collected and washed with alcohol to yield 0.28 g. (81%) of uracil XVI, m.p. 215" dec. Recrystallization from methanol-ether gave a pure sample, m.p. 216-217" 258 mp ( e 9,020); pK'a 9.9 (in 507% dec.; [ a I 2 5 ~ 71.3 (c 1.05 in H2O); : : :A MeOH). Anal. Calcd. for CIZHI??IT307: C, 45.71; H, 5.44; K, 13.33. Found: C, 45.45, 45.47; H, 5.66, 5.57; Y, 13.11. l-(2-Deoxy-2-amino-~-~-g~ucopyranosy~)-uracil (XV).-A solut,ion of 1 g. of nucleoside I V in 10 ml. of water and 10 ml. of concd. hydrochloric acid was refluxed for 10 hr. The solution was then evaporated to dryness in va,cuo. The residue waa dissolved in a small volume of methanol and deposited crystals on standing. These were collected, washed with a little methanol and then with ether t o give 0.615 g. (91%) of I-(2-deoxy-~-amino-p-D-glucop~ranosyl)-uracil hydrochloride, m.p. 275-278" drc. Itecrystallization from aqueous methanol gave an analytical sample, m.p. 280" dec., with blackening above 240'; [.I% + 257 mp ( L 9,790): pK'a 6.2>10.1 (in 50% MeOH). 40' (c 1.2 in H20); Anal. Calcd. for CloH&lS:&: C, 38.77; H, 5.21; S , 13.57; C1, 11.45. Found: C, 38.92, 39.10; H, 5.21, .5.56; S , 12.85; C1, 11.39. The same compound could also he prepared by vigorous acid hydrolysis of 1(3,4,6 - tri - 0 - acetyl 2 deoxy - 2 - carbomethoxyamino - P - D - glucopyranosy1)4-inethoxy-2-pyrirnidinone ( V I ) i n 53 yield, m.p. and mixture rn.p. 280" dec. The infrared spectra of the two preparations were superimposable. A suspension of 0.68 g. of this hydrochloride in 6 ml. of ethanol containing 0.6 ml. of diethylamine was shaken for sweral hr. The mixture was filtered and t,h[, precipitate again shaken as a siispension in 2 nil. of ethanol containing 0.2 ml. Of diethylamine. Filtration and washing with ethanol afforded 0.495 g. (91%) of 1(2-deoxy-2-amino-~-~-g~ucopyranosy~)-uraCi~ ( Y l r ) ,m.p. 239-240" dec. Onc crystallization from moist methanol-ether gave needles, m.p. 241-2.12" dec.; ED 15.66" (c 1.1 in H20); AA'eo,Hk.H20 259 mp ( e 9,870); pK'a 6.1, {I.!)(in SOYc MeOH); pK'a 5.87, 9.17 (in water). Anal. Calcd. for C I O H I S N ~ OC,~ : 43.93; 11, 5.53; N, 15.36. Found: C, 44.16; H, 5.48; N, 15.34. The picrate was formed by warming the hydrochloride with alcoholic picric acid. Crystallization from methanol-ether gave yellow cubes, m.p. 254-255O dec., [ a l Z 5 ~f 25.1 (c 1.1 in H20). Bnal. Calcd. for C I ~ H I ~ N ~C,O 38.25; ~ ~ : H, 3.61; N, 10.73. Found: C, 38.47; H, 3.58; N, 16.72. ~.(2-Deoxy-2-carbobenzyloxyamino-~-~-glucopyranosyl)-uracil (XVII).-A POlution of 0.28 g. of nucleoside V in 20 ml. of warm nicthanol wa$ added to 7 nil.
+
+
- -
+
of ttlcohul containing 0.5 g. of hydrogen chloride. After hiving for 24 hr. :tt room temperature, the solution was evaporat,ed to dryness. The residue was dissolved in 5 ml. of alcohol m d on standing deposited 0.12 g. (58c/;)of crystals, n1.p. 225-226" dec. Recrystallization from niethanol-ether gave a sample of the same inelting point; [ c Y ] ~ ~-D 34' ( c 0.78 in H?O); Xh:2gi 258 mM ( e 9,410). A n ~ l . Chlcd. for Cjt.H?iSZOa: C , 53.0;; H,5.20: K , 10.32. F O U I I ~ : 5:3.25, 52.73; H, 5 . 3 , 5.05; S , 10.L7S. 1-(3,4,6-Tri-~-acetyl-2-deoxy-2-carbobenzyloxyamino-~-~-glucopyranosyl~1uracil (XXI).-A solution of 1 g. of nucleoside V TWS converted t o 1-(2-deosy-2carbobenxylo~g-an~~nc~-~-~-glucop~ra~nos~l)-urac~l :is above. The crude product \vas acctylated without prificat,ion by heating wit,h 3 nil. of :icetic anhydride at IOUo for 4 lir. The solut,ion was evaporated to dryness in I : U C U O . Addition of alcuhul to the residue afforded 0.6 g. (62%) of crystdline XXI, ni.p. 190-191". Recrystallization from alcohol-ether yielded shining clusters of needles, m.p. lYl-102°; [0(]251) - 20.3" ( e 0.95 in CHC1,): XF$,I:! 236.5 n i (e$IJ!)O): ~ pK'n 10.7 (in 50cjc MeOH). .And. Cdcd. for C21H&~Oll: 0, 54.04: H, 5.10; S . 7.h8. Found: C',, x ~ z5 ,u i ; I-I, 3.23: 5.25; N, ;.xs. AL more direct and sat,isfactory preparation of this compound ( S X I ) was as follows: solut,ion of 1 g. of nucleoside 1 ' in 50 Ink. of chloroform and 5 nil. of acetic anhydride was saturated with dry hydrogen chloride gas a t room t,eniperahre. ilfter leaving overnight, the solution was evaporated to dryness in. ucicrLo at roc1111 temperature. The residual gum was dissolved in alcohol and ether added t u it, t o the point of crystallization. Pi1tr:Ltion and washing with ether gave 0.94 g. ('375; ) of uracil XXI, m.p. and misturci m.p. 1%-1 !IO0. 1- (3,4,6-Tri-O-acetyl-2-deoxy-2-amino-~-~-glucopyranosyl ,-uracil(XXv).--X solution of 1 g. of carbobenzylosynucleoside 9x1 in 40 nil. of purified, acid-free dioxane was subjected to hydrogenation at :ttmospheric pressure using 0.6 g. of l0yopalladium-on-charcoal as a catalyst,. At the cxnd of 2 hr., after an uptake of 0.9 mole of hydrogen, absorption slackened considerably. The reaction was stopped at this stage. The mixture was filtered and the filtrate evaporated in vucuo at room temperature. Since nttcnipts to tallize the aininoiiiicleositl(. using an c.thcr holutio~ioi failed, it was convertrd t o the hydrochloride s hydrogen chloride. The precipitated salt FWF collect,ed, wv:tshetl with ethor : i n t i dried t o yield 0.72 g. (SSyO) of white solid, n1.1). 230" dec. ('rgstallizatiou frorii methanol-ether gave an analytical saniple, 111.1,. 232.O der.; [CY]% 29.9" " 'Ic1 56 inp ( e 9,110); pK'a 1.9, !).6 (in 301;; MeOK). (c 0.66 in H?O); A n d . Calcd. for ClsH Kz09: C, 44. io; t L ;.O
) 01' t h c b iiirii~~t1iyl:~riiiri to reniove cwess siilfoiiic*:wid, l i tained as orango Iieedles, ii1.p. 215- 21 6" dw,, 111.1). iiiitI(:1,ressc~lk)y :tcliiiixturc~ with a n authentic specimen.; 1-(2-Deoxy-2-dimethylam~no-~-~-glucopyranosy~)-cytos~ne [XIV)..-.4 sohtion of 0.35 g. of nucleoside SXTS in 20 nil. of alcohol saturated with ninnioiii:i nt 0' was heated in a sealed tube for 40 hr. The tube was opened and the solution evaporated to dryness. The residual gummy base \voiild not cxystallizc~ und was converted int,o the sulfate by addition of 2 drups of concd. sulfuric acitl t o its alcoholic solution. The salt was complrtely precipitated I J addition ~ oi ether. Crystallization front aqueous alcohol afforded 0.285 g. (5Xjf I of tliv sulfate nionohydrat,e. m.p. 240" dec. wit,h hlackeniiig above 230"; ~ o ] ? ~ I I 56.7" (c 1.05 in H10):'=.:A: Firr272.5 nip ( e 12,200)! pk":i 3.1. 5.5 ( i n 50( Anal. Chlcd. for C1?H?J~098.H20:49C,, 34.61 ; H. 5.Sl: h-, 1:{,46: rieiit>ralizationequivalent, 208.2. Found: C, 34.36, :34.$18; 11, 5.B:i. 5 . G : S. 1:{,55. I3.:%0;neutralization rquivaient, 202.3. Periodate Fission of 1-~2-Deoxy-2-dimethylamino-~-~-glucopyranosyl~-cytosine (XIV),---4 solut,ion of 0.243 g . of t,he sulfat,cl rnoiiohyt1r:ttc~JKW nciitralizc~tl with h r i u n i hydroxide :inti the base isolated as ;i gun1 1)y t h c iisu:tl procwiurc~. 'This was dissolved in 10 nil. of water contairiirig 0.S0 g. of sodium iiietupc,ri[,il:ii,~~ rind 0,:31 g. of picric acid. .\fter 70 hr. at room temperuturr, tlic picrate salt, \V:I< r:~)llectedand witshed wit,h a little x:tter t,u nft'ord 0.14 g. (51hemelting point rose to 215-21 (io 55" (c 0.67 ill pyridinej. Identity was est.ablished i)y niist,iirc. de(&.; [ a ] % melting point, with the picrate of dialdehyde XXIII from the fissiori of cyticti~io"~ and the superimposability of their infrared spectra. A n a l . Calcd. for CjjHI.iS&?: (', Y8.31 ; H, 3.00. Foiintl: C, 3ii.lS, :i7 H, 3.19, 3.17. The mother liquors from this reitction were combined and passed through :I 15 x 2 cm. colunin of Dowes 50 (25-50 mesh). -4fter washing with water uiitil the eluate was neutral, t,he column was rinsed with 125 ml. of 2 S hydrochloric. acid and the e1u:tte evaporated in uucuo to dryness. The residue was dissolved in a little water and after addition of 20 ml. of 1056 potassium hydroxide solution it, was d i d l e d into an aqueous solution of 0.175 g . of p-hydrox?.azobenzrnr-~'julforiic acid. Evaporation CJf this solution and crystallization from :dcuhol. :ifforded 90 mg. (50yo)of t,he dimethylamine salt, m.p. and mixture m.p. uitli :iut,hentic specimen,&216-217" dec. 1dentit.y was further confirmed by compnrisoii of infrared and ultraviolet absorption spectra and X-ray powder patterns. 1- (3,4,6-Tri-O-acetyl-2-deoxy-2-dimethylam~no-~-~-glucopyranosyl) -4-acetamido-2-pyrimidinone (XXVII). A. From 1-(2-Deoxy-2-dimethylamino-p-uglucopyranosy1)-cytosine (XIV).--.4n aqueous solution of 0.27 g. of t'he siilfatc monohydrate of nucleoside XIV was neutralized with aqueous barium hydroxide. The free base, isolated by the usual procedure, was then refluxed with 5 ml. of acetic anhydride and 2 nil. of pyridine for 5 min. The ~olut~ion was allowed t,o stand overnight and then evaporated t o dryness in. vucuo. The residue was st,irrcct wit,h 5 ml. of water and extracted with rhiuroform. The chloroform layer was
+
-+
(49) .&iter drying to constant weight in ziacuo a t loo", the sample stili had a correct analy.1r:xcept f o r a hpmihydrate: Calcd.. C , 3.5.47; H , 5.71: N.13.80. Found: C , 35.18, :35.$4: H T,.B?, 5.77; N, 13.43, 13.58.
Novmber, 1962
SYXTHESIS OF AMMINOSUGAR NUCLEOSIDES
1147
dried over sodium sulfate and evaporated t o dryness in vacuo. The residue was triturated wit.h ether and the crystalline solid collected by filtration. There was thus obtained 0.27 g. (89%) of the acetyl derivative, m.p. 258-259'. Crystalliaation from 1-butanol gave needles, m.p. 259-260"; [(YIz5D f 73.4" (c 0.92 in HC' (after HC1 (initial) 251 mp ( e 15,700), 297.5 mp ( e 7,540); CHCl,); A&x: 24 hr.) 270.5 nip ( e 12,410); pK'a 2.8 (in 50% MeOH). Anal. Calcd. for C20H25X409: C, 51.27; H, 6.02; X, 11.96; 0, 30.74; 4 CH,CO, 36.77. Found: C, 50.90, 51.04; H, 6.32, 6.14; S , 11.75, 12.14; 0, 30.34, 30.60; CHSCO, 36.40. The hydrochloride was prepared in chloroform solution by the addition of ethereal hydrogen chloride. The crude conipound was hygroscopic, shrank above 150" and melted a t 180" dec. By addition of sodium bicarbonate to an aqueous solution of the salt, the base could be recovered unchanged. An attempt to crystallize t,he hydrochloride from ethanol-1-butanol afforded, after several days, whit,e crystals, m.p. 228-229" dec. This compound however had an ultraviolet, absorption maximum at 276 mg (E: 222), indicating considerable hydrolysis of Lhe acetylcytosine moiety to free cytosine.'5 B. From 1-(3,4,6-Tri-0-acetyl-2-deoxy-2-am~no-~-~-glucopyranosyl)-4-acetamido-2-pyrimidinone (XXVI).-A solution of 0.2 g. of carbobenzyloxyaminonucleoside VI11 in 10 ml. of dioxane was subjected t o hydrogenation using 0.1 g. of 10% palladium-on-charcoal as a catalyst. The mixture was filtered without isolating the aminonucleoside XXVI. Aqueous formaldehyde solution (0.3 ml.) was added to the filtrate and reductive methylation carried out, using 0.15 g. of freeh catalyst. In the course of 20 hr., about 1.6 moles of hydrogen (theoretical, 2 moles) was absorbed. Since no more hydrogen was taken up, the mixture was filtered and the filtrate evaporated in vucuo to dryness. The residual syrup was triturated with 5 ml. of ice u-at,er, whereupon crystallization occurred, d f t e r cooling a t 5' for 1 hr., the mixture was filtered and the residue washed with a little water t o give 50 mg. (31%) of product, m.p. 255-257". After one crystallization from methanol-ether, t.he dimethylaminonucleoside XXVII was obtained as needles, m.p. and mixture m.p. 259-260'. The ultraviolet and infrared ahsorption spectra of the two samples were identical. Ammonolysis of 1-(3,4,6-Tri-O-acetyl-2-deoxy-2-d~methylamino-~-~-glu~0pyranosyl) -4-acetarnido-2-pyrimidinone (XXVIL).-A solution of 35 mg. of nuclcoside XXVII in 5 ml. of alcohol saturated a t 0" with ammonia was heated in :I sealed tube a t 100-110" for 48 hr. The solution was evaporated to dryness and the gummy base converted into the sulfate salt. Crystallization from aqueous alcohol gave 25 mg. (80%) of product, m.p. 234-236" dec., undepressed by admixture with the sulfate salt of nucleoside XIF'. Identity was confirmed by comparison of infrared and ultraviolet absorption spectra. 1-(3,4,6-Tri-0-acetyl-2-deoxy-N-methyl-deoxy.~-methyl-2-acetam~do-~-~-glucopyranosyl) -4methoxy-2-pyrimidinone (XXXII).-A solution of 4 g. of pentaacetyl-N-methyl-aglucosamine41in 20 nil. of acetic anhydride was cooled t o 0' and saturated with dry hydrogen chloride gas. The solution was kept a t room temperature for 24 hr.: then cooled t o O", saturated with hydrogen chloride gas and set aside for another 3 days. The solution was then evaporated in uucuo, the residue dissolved in chloroform and shaken successively with ice water, cold sodium bicarbonate solution and again ice water. The chloroform layer was dried and evaporattsd in ~ m c u o . Tho residual syrup was trituratcd with ether, whcrcupon a crystalline solid separated. This was collectcd and washed with a little ether to give 1.6 g., lV
C. L. STEVEM A K D IC. XAGSRAJAS
1148
Vol. 5
n p . 158-15Yo, identical with the starting pentaacetate. The ether filtrate, on evaporation, left a halogen-containing syrup which was then refluxed with 3 nil. of 3,4-dimethoxypyrimidine and 15 ml. of benzene on a steam bath for 40 hr. The solution was decanted from the black tar that had formed and subjected t o chromatography over a column of 50 g. of alumina. After removal of excess unreacted dimethoxypyrimidine by elution from alumina with benzene, the material eluted by t,he first 50 ml. of benzene containing 0.5% alcohol was discarded. The nucleoside was obtained from the eluate of the second 50 nil. portioii of benzene containing 0.5':5 alcohol. Evaporation of this fraction and rubbing with ether afforded approximately 0.1 g. of needles which were recrystallized from alcohol-ether, m.p. 227-229"; 276 mp ( e 4,500). Anal. Calcd. for C ~ O H U S & : C, 51.16; II, 5.80; S , 8.95. Found: C, 50.74; H, 6.12; N, 9.54. 1,3,4,6-Tetra-0-acetyl-2-deoxy-2-dimethylamino-p-~-glucopyranose (XXX1j.-To a solution of 1.91 g. (0.5 mmole) of ~-1,3,4,6-tetra-O-acetylglucosaminehydrochloride (XXX)'&in 10 nil. of water was added 2 g. of sodium acetate. The base which was liberated Kas extracted with 3 X 25 ml. of chloroform. The chloroform ex;tract W R Y evaporated in vacuo. The residue was dissolved in 20 1n1. of dioxane containing 3 nil. of a 35% aqueous solution of formaldehyde. Hytlrogenation was carried out at atmospheric pressure using 0.5 g. of lo';;,; palladium-on-charcoal aa a catalyst. Uptake of hydrogen ceased a t 2 moles at the cxd of 15 hr. The mixt,ure was filtered and the filtrate evaporated in O ~ C L L O . The syrupy residue, smelling strongly of formaldehyde, was dissolved in water and extracted with chloroform. The chloroform layer was dried over sodium sulfatc and evaporated. Addition of &hereal hydrogen chloride to an ether solution of the residue gave the crystalline hydrochloride salt, 1.32 g. (G2yO), m.p. 167-168". liecryst,allizatioii from alcohol-ether gave colorless prisms, m.p. 1GX-lTO" dcc., [aIn5o 28.5' (c0.6 in H20); pK'a4.2 (in50% MeOH). Anal. Calcd. for Ci6H&1?;O9: C, 46.67; H, 6.37; C1, 8.61. Found: C , 46.54,46.3T; H, 6.31, 6.34; Clj 8.54.
+
Acknowledgment.----Theauthors wish to thank Dr. J. 31. Cniideiihelt a i d asswiatw at Parkc, Davis & Co., for the spectral aiid t8itrat ion data published herein and C. E. Childes a i d associates of th(: same company for most of' the analytical determinations.
