Soveniber 1967
1lti5
SOTES
Anal. Calcd for C~isIILIN6019’0.6Hz0: C, 34.12; I T , 3.97; [in mp ( e X 0.1 S HCl 278 (15.4), 0.1 mg (314,); A,, N, 14.92; P, 5.50. Found: C, 34.37; I€, 4.31; N, 14.83; P, -YKaOH 276 (11.5); v (in cm-1) 3410 (OH), 3070, 2930, 2810 5.41. ICH). 1690 ( C a ) , 1615, 1445 (C=C, C=N), 1035 (P-0-C); MI,, (pH 4.4) 0.68, (pH 7.2) 0.41. Acknowledgments.-The authors are indebted t o dnal. Calcd for C18HzlBrzN4012P:C, 31.97; H, 3.18; S , Dr. W. .J. Barrett arid members of the Analytical and 8.29: P. 4.58. Found: C, 31.89; H, 3.23; S , 5.13; P, 4.39. 2’-D~oxy-5-iodouridylyl-(5~-5~)-2‘-deoxy-5-iodouridine (6c). Physical Chemistry Division of Southern Research -To 711 mg (1.80 mmoles) of 4c in 10 ml of anhydrous dioxane Institute who performed most of the microanalytical was added 231 mg (0.90 mmole) of p-nitrophenylphosphorodiand spectral determinations arid to Dr. G. J . Dixori arid chloridate followed by 1.95 ml of dry pyridine. The solution was J. 11. Thomson for the biological results reported. worked up as described above for 6b. The thin layer chromatography purification of the product was also run in the same way iising 3: 1 CHC13-14eOH as the developing solvent to give a white [in mp ( e X 0.1 HC1 287 wlid: 188 mg (277,); A,, (12.9), 0.1 iV NaOH 280 (10.1): v (in em-l) 3410 (OH), 3070, 3’-Deoxynucleosides. V. 2940, 2810 (CH), 1680 (C=O), 1610, 1443 (C=C, C=K), 3’-Deoxy-2-fluoroadenosine 1O.X ( P a - C ) ; MI, (pH 4.4) 0.62, (pH 7.2) 0.33. *Ana/. Calcd for ClRH2~I2X4Ol2P:C, 28.07; H, 2.7;; Y, 7.2X; l’, 4.02. Found: C, 28.03; H, 2.8!); K, 7.08: P, 3.91. 1-(2,3,-0-Isopropylidene-~-~-ribofuranosyl)-6-azauracil (7).“--~To 13.5 ml of 2,Zdimethoxyproparie in 300 ml of dry acetotic was added 18.4 ml of 70% HC104.12 l f t e r 5 mill, 10.0 g (40.8 mmoles) of 6-azauridine was added. The resulting mixture was stirred for 50 min a t room temperature, neutralized by the addition of 21 ml of pyridine, and evaporated t’o dryness in vacuo. The residue was shaken with a mixture of 100 ml of CHCl, and 30 ml of H20. The aqueous layer was again extracted with 100 The effectiveness of 3’-deoxyadenosine (1) (cordyml of CHCI,. The CHC13 solutions were combined, dried cepin) as a cytotoxic agent is diminished by its rapid (hlgS04), and evaporated to dryness in vacuo. An acetone conversion into inactive 3’-deoxyinosine (11) through solution of the residue was also evaporated to dryness. This the action of adenosine deaminase.l On the other residue was dissolved in 25 ml of acetone. The addit’ion of 130 ml of cyclohexane produced an oil that crystallized upon seedhand, 2-fluoroadenosine (111), 2 a potent cytotoxic ing; yield 5.45 g. The aqueous layer from above was diluted wit’h enough concentrated NHaOH to give pH 2 and again extracted P with CHC13 (two 100-ml portions). The CHCl3 extracts were worked up as described above. A second crop of crystalline R I material weighing 2.30 g was obtained: total yield 7.73 g (66.5%); [in mp ( e X 0.1 AV mp 141-142’ (lit.6 141-142”): A,, HC1 260 (6.48), 0.1 X NaOH 234 (7.25): v (in cm-l) 3540 (OH), 3140, 3100, 2990, 2820 (CH), 1i25 (C=O), 1670 (NH), 1585 (sh) (C=K). 6-Azauridylyl-(5’-5’)-6-azauridine (lo).-To 268 mg (0.94 mmole) of 7 in 5 ml of anhydrous dioxane was added 132 mg R“ OH (0.52 mmole) of p-nitrophenylphosphorodichloridate followed by I, R = NH,; R’ = H; R” = H 1 ml of dry pyridine. The mixture was stirred at room tempera11, R = O H R = H;R” = H ture for 2 days. Examination of an aliquot by thin layer chro111,R = NH,; R’ = F; R” = OH matography indicated incomplete reaction. Therefore, another IV, R = NH2;R’= F; R = H 10 mg of p-nitrophenylphosphorodichloridatewas added. After V, R = R’= NH2;R”= H another 24 hr, the solution was diluted with 50 ml of H,O and ext,racted with three 150-ml port,ions of CHC1,. The CRCI, itgcnt ,? is relatively inert to the action of adenosine was dried (SlgS04) and evaporated in vacuo. The p-iiit,rophenylbis(2,3-0-isopropylidene-6-azauridine)5’,S”’-phosphate (8) was deaminase. It appeared that iricorporation of a thus obtained as a white, powdery glass; yield 262 mg (74%,). fluorine atom a t the 2 position of 3’-deoxyadenosine Without further purification, the glass in 10 ml of 0.3 S would give a derivative, 3 ’-deoxy-2-fluoroadenosirie KaOH was left 2 hr at room temperature. The solution was (IT), which would be stable to adenosine deaminase and neutralized by stirring it with Rexyn RG 50 ( H ) ion-exchange might retain the biological properties of 1 . 4 For the resin. The resin was removed by filtrat,ion, and the filtrate was washed several times with ether to remove p-nitrophenol. Evaposynthesis of IV, 2-amino-3 ‘-deoxyadenosine (V)5 was ration of the aqueous solution to dryness in vaczto gave a gummy subjected to a modification of the procedure2 used for residue which became a white solid upon trituration i n EtOH. the preparation of 111, which involved the selective The EtOH was removed by evaporation. The process was diazotization of the 2-amino group in the presence of repeated several times giving 167 mg of bis(2,3-0-isopropylide~ie6-axauridine) 5’,5”’-phosphate (9) as a white solid which mas fluoroboric acid. Crystalliiie 3’-deox~r-2-fluoroadenodissolved in 50 ml of H20. The sollition (pH 2 ) was refluxed for sirie (IV) w i \ ot)taiiied i n 18% yield. 1.5 hr, t,hen evaporated to dryness in V U C I I O . The 1,esidiie was 111the preseiice of calf intestine adenohine deaminase, triturated in EtOH, and the Et,OH was removed by evaporation. I V was riot meaburably deaminated under conditioris The process was repeated several times. The white, powdery which accomplished complete deamination of adenosirie. glass was dried for 18 hy at 100” (0.07 mm) over P,O,: yield 144 [ i n mp (E X 0.1 A’ HCI 261 (12.6), mg (497,): A,,, 3 ’-Deoxy-2-fluoroadenosine did not inhibit the deamina0.1 -1-XaOH 233 (14.3); v (in cm-’) 3430 (broad) (OH), 1740 (pH 4.4) 0.80, (pH (sh) and 1690 (C=O), 1025 (P-0-C); (1) H. Klenow, B i o c h i m . B i o p h y s . A c t a , 76, 347 (1963). 7.2) 1.07. (2) (a) J. A. Montgomery and Ii. Heason, J . Am. Chem. S o c . , 79, 4559 The atialytical sample was obtaiiied by chromatography on a (1957); (h) ibid., 8’2, 463 (1960). SIalliiickrodt silica gel t i o . SG7 plate using BuOH-AcOH-Hz0 ( 3 ) (a) 0. P. Chilson and J. R. Fisher, A r c h . Biochem. B i o p h y s . , 102, 77 (1963); H. P. Barr a n d G. I. Drummond. Biochem. B i o p h y s . Res. Conirnun., ( . j : 2 : 3 ) iis the eluent. The band obtained was removed from ar, 584 (1966). the silica gel with hot hIeOII. The solid was dried a t 100’ (4) S. Fredericksen, B i o c h i m . B i o p h y s . A c t a , 76, 366 (1963), has reported (0.07 nim) over P205for 18 hr.
13)
(12) J . .‘I. Zderic, J . G. N o f f a t t , I). K a u , K. Gerzon, and K.E. Fitagibbon, J . M e d . C h e m . , 8 , 275 (1965).
t h a t cordycepin 1-oxide is resistant t o enzymic deamination a n d t h a t enzymic reduction provides for in s i t u release of cordycepin. (5) E. Walton, F. W.Holly, G. E. Boxer, R. F. N u t t , and S. R. Jenkins, J . Xed. Chem., 8, 659 (1965).
