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TETRAHEDRON

Pergamon Tetrahedron 57 (2001) 9163±9168

Synthesis of 1 0-aza-C-nucleosides from
(3R,4R)-4-(hydroxymethyl)pyrrolidin-3-ol
Vyacheslav V. Filichev and Erik B. Pedersenp
Department of Chemistry, University of Southern Denmark, Odense University, DK-5230 Odense M, Denmark
Received 23 April 2001; revised 17 August 2001; accepted 6 September 2001

AbstractÐPyrimidine 1 0 -aza-C-nucleosides are synthesised by the fusion of 5-bromouracil, 5-bromocytosine and 5-bromoisocytosine with
(3R,4R)-4-(hydroxymethyl)pyrrolidin-3-ol in 40±41% yield. A homologue of 1 0 -aza-C-uridine is obtained in a Mannich reaction in 65%
yield by treatment of the azasugar, paraformaldehyde and uracil. N-Alkylation of (3R,4R)-4-(hydroxymethyl)pyrrolidin-3-ol with 6-chloro-
methyluracil gives the 6-regioisomeric homologue. (3R,4R)-4-(Hydroxymethyl)pyrrolidin-3-ol is synthesised in 25% overall yield from
diacetone-d-glucose via 3-C-(azidomethyl)-3-deoxy-d-allose which is subjected to an intramolecular reductive amino alkylation reaction to
give (3R,4S)-4-[(1S,2R)-1,2,3-trihydroxypropyl]pyrrolidin-3-ol followed by Fmoc protection, oxidative cleavage of the triol group with
further reduction of the obtained aldehyde and subsequent deprotection of the nitrogen atom. q 2001 Elsevier Science Ltd. All rights
reserved.

1. Introduction ing to the procedure of Phillips for condensation of amines
with 5-bromouracil.9 According to this procedure the
Since the discovery of pseudouridine (C-uridine) 1 (Fig. 1) racemic 5-[3-hydroxy-4-(hydroxymethyl)pyrrolidin-1-yl]-
in nature in 1957,1 a considerable number of C-nucleosides uracil was prepared as a 1 0 -aza analogue of C-uridine.8
have been synthesised and found application as antibiotics Racemic trans-3-hydroxy-4-hydroxymethyl-N-(6-uracilyl-
and potential anticancer and/or antiviral agents.2 Due to methyl)pyrrolidine, that is considered an analogue of
their structural relationship C-nucleosides can be incorpo- inosine because it mimics the pyrimidine ring, could
rated into DNA/RNA instead of the natural occurring likewise be obtained by condensation with 6-(halogeno-
nucleosides. On the other hand the modi®cation of the methyl)uracil.10
glycons is one of the main directions in the synthesis of
nucleoside analogues. Among the possibilities are nucleo-
sides with replacement of the natural glycons with poly-
hydroxylated cyclic amines known as azasugars or
iminosugars. These sugars are also of interest as inhibitors
of various glycosidases.3 3 0 -Aza-3 0 -deoxythymidine carbo-
analogue4 and the pyrrolidinyl analogues of 2 0 ,3 0 -dideoxy-
cytidine5 (2) were among the ®rst representatives in this
area. Lee and co-workers6 have published the synthesis of
1 0 -aza carbacyclic thymidine analogues 3 from the racemic
1-benzyl-4-(hydroxymethyl)pyrrolidin-3-ol.7

Recently our group has presented a new class of nucleo-
sides, called aza-C-nucleosides, in which a carbon in the
heterocyclic base is linked to the nitrogen of the secondary
cyclic amine.8 For developing the synthesis of aza-C-
nucleosides, the challenge is to synthesize pure enantio-
meric azasugars. These can then be condensed with
5-bromopyrimidines to produce aza-C-nucleosides accord-

Keywords: azasugar; iminosugar; reductive amination; (3R,4R)-4-(hydroxy-
methyl)pyrrolidin-3-ol; pseudonucleoside; aza-C-nucleoside.
p
Corresponding author. Tel.: 145-6550-2555; fax: 145-6615-8780;
e-mail: ebp@chem.sdu.dk Figure 1.

0040±4020/01/$ - see front matter q 2001 Elsevier Science Ltd. All rights reserved.
PII: S 0040-402 0(01)00920-6

145±1508C. furanose and 2-deoxy-d-heptofuranose in a one-pot reaction by a catalytic reduction reaction of amino group precursors in aldosugars (3-C-cyano-3-deoxy-d-ribo-pentofuranose. Then. 10 min. (d) Amberlite IR-120(H1).15 which in this investigation was prepared in situ. the Wittig reac- 10 min. (f) FmocCl/dioxane/10% aq. In this synthetic route the puri®cation of products using silica gel column chromatography is neces- Scheme 2.11 We the end low overall yields (maximum 11%) were obtained have synthesized azasugar analogues of 2-deoxy-d-hexo. NaHCO3.4R)-4-(hydroxymethyl)pyrrolidin-3-ol (12) in 67% yield from 9. Compound 6 was prepared in three steps from 513. of dimethylammonium azide11. 2.13 Scale up of the Wittig reaction to 30 g led to a decreased yield (30%). (g) (1) NaIO4.11 Multistep reac- merically pure aza-C-nucleosides utilising a synthetic tions including multiple chromatographic puri®cations were route for the required azasugar which is more easy and needed in order to obtain the amino precursor sugars and in straightforward than the ones previously published. 10 min. Reagents and conditions: (a) 5-bromouracil.2:5. reduction with sodium borohydride and deprotection with NEt3 in acetonitrile11 afforded (3R.6-di-O-isopropylidene-a-d-ribo-hexofuranos-3-ulose. We found it attractive to synthesise (3R. The best yield (57%) was obtained from 20 g of 1. fusion. Filichev. practically useful than the ones previously published. V. In order to avoid contamination with sulfur compounds which later on could poison the catalyst during the reductive amination. (3) MsCl/Py. (2) CH3P(C6H5)3Br/n-BuLi/THF. Fmoc protection of the primary amine followed by oxidative cleavage of the triol group in the corresponding Fmoc-azasugar 10 gave 11. (e) H2/200 psi/Pd±C/H2O.9164 V.14 and was then converted to compound 7 by treatment with 1. Reagents and conditions: (a) (1) CrO3/Py/Ac2O/CH2Cl2. of the azasugars.4R)-4-(hydroxy- methyl)pyrrolidin-3-ol (12) through 3-C-azidomethyl-3- deoxy-d-allose (8) starting from diacetone-d-glucose (4) (Scheme 1). (c) (CH3)2NH´´ ´HN3/DMF. 145±1508C. 145±1508C. Deprotection of compound 7 under acidic conditions afforded the azido sugar 8 in an overall yield of 39% from 4. (b) 5-bromocytosin. Pedersen / Tetrahedron 57 (2001) 9163±9168 Scheme 1. (b) (1) BH3/THF. (2) NaOH/H2O2. (h) NEt3/acetonitrile. (2) NaBH4. molecular reductive alkylation reactions. E. fusion. . synthesis of azide) and this makes this scheme more cytosin.5 equiv. This paper describes the synthesis of several enantio. (c) 5-bromoiso. B. sary only in three steps (oxidation by CrO3. The required azasugar 9 was obtained as a brown oil in 80% yield by a reductive amination reac- tion using hydrogen over 10% Pd/C in water in an autoclave for 16 h. tion. fusion.11 the methylene derivative 5 was obtained from diacetone-d-glucose (4) by oxidation with a complex of CrO3/acetic anhydride/pyridine12 followed by treatment with the Wittig reagent MePPh3Br/n-BuLi in dry THF. Results and discussion 3-C-azidomethyl-3-deoxy-d-ribo-pentofuranose and 3-deoxy- 3-C-nitromethyl-d-allose) followed by an in situ intra.

fusion. 117. The NMR spectra were con®rmed by comparison with calculated spectra using ACD/HNMR predictor and ACD/ CNMR predictor. Uracil was reacted according to the procedure of Motawia et al. 5-bromocytidine could be converted to the corresponding 5-aminocytidine in liquid ammonia. our attempts to obtain the aza-C. However. 1 0 -Aza-C-nucleosides 13±15 were obtained by the fusion To obtain 1 0 -aza-C-cytosine analogues we used the same of the azasugar 12 and 5-bromopyrimidines in 40±41% methodology.4) in 13 and 14. (CH2O)n.3.6-trimethyluracil with compound 9.20 They found that re¯uxing of 1. respectively. Conclusion gel column chromatography and subsequent recrystalliza- tion (Scheme 2). 13. yield.95 ppm compare to the NH signal at 10. The homologues of 1 0 -aza-C-uridine 16 and 17 primary and secondary amines have been examined by were obtained from the azasugar 12 in the Mannich reaction . 24 h.6-trimethyluracil-6-yl)pyrrolidine (18) and time and increasing the polarity of the solvent.9 5-[(3R. Excess of pyrrolidine 12 was also isolated In the present investigation we have devised a simple way of from the residue and this is important because 12 is not synthesising one of the enantiomers of a 1-aza analogue easily available. An interesting property of the nucleosides 13±15 is their potential ability of existing both as a and b-anomers due to ¯ipping of the ring nitrogen.1. B. Spectra for all the possible tautomers of the compounds 13±15 were calculated and the tautomers in Scheme 2 are those giving the best ®t with the calculated ones. fusion of a any conversion of 18. respec- tively. MeOH. large excess of an amine with a bromoheterocycle. neither (3R. (b) 6-chloromethyluracil (for 17). 131. 1008C.17 The fusion of the azasugar 12 with 5-bromocytosine and 5-bromoisocytosine at 145±1508C for 10 min led to the 1 0 -aza-C-isocytosine 14 and 1 0 -aza-C-cytosine 15. This is explained by a shift in the tautomeric form of compound 15 to the OH-form and this group appears in the 1H NMR spectrum at 6. Pedersen / Tetrahedron 57 (2001) 9163±9168 9165 Phillips16 in 1953 and less reactivity of 5-bromoisocytosine was found when compared with 5-bromouracil.4R)-3-Hydroxy-4-hydroxymethyl-N-(6-uracilylmethyl)- products of coupling nor starting azasugar were observed pyrrolidine (17) was prepared in 50% yield by N-alkylation in the black reaction mixture. E. The reaction of 5-bromoisocytosine with yield. re¯ux.18 A characteristic feature of the cytosine derivative is the lower ®eld signals of C-5 (128. This was con®rmed by NOE spectra. (c) guanidine. However.19 with paraformaldehyde and the secondary amine 12 to give the Mannich base 16 in 65% yield.3 ppm in compound 14.2.3. On irradiation of H-6 3±4% NOE was observed in H-2 0 and H-5 0 for both b and a hydrogens in compound Scheme 3.6) and C-6 (125. guanidine for 6 h led to the 6-methylisocytosine in 45% nucleoside 13 from 12 under the same conditions also failed. heating.e. of 2-deoxy-d-ribofuranose from diacetone-d-glucose (4). 10 h. Reagents and conditions: (a) uracil. According to the literature.4R)-3-Hydroxy-4-(hydroxymethyl)pyrrolidin-1-yl]- uracil (13) was isolated in 40% yield after fusion of 3 equiv.3-ambident nucleophile using the procedure Hirota et pyridine and in alcohols in contrast to the partly soluble al. The sugar 12 dissolves completely in by a 1.8) in 15 compare to the signals of C-5 (120. Filichev. rt. both in 41% yield. of the azasugar 12 and 5-bromouracil followed by a silica 3. 6-chloromethyl-1. but as earlier observed. i. DMF). enantiomerically pure 1 0 -aza-C-uridine from 5-bromouracil and azasugar 9 under re¯ux in pyridine for 24 h.4R)-3-hydroxy-4-hydroxy- The problem could be solved by decreasing the reaction methyl-N-(1.8 DMF We attempted the synthesis of the guanosine analogue 19 by is undesirable as solvent because of side reactions with nucleophilic displacement of the N1 ±C2 ±N3 fragment in 18 dimethylamine. The presence of the vicinal of azasugar 12 with 6-chloromethyluracil and HuÈnig's base hydroxyl groups makes the structure 9 too sensitive for in MeOH. EtOH. (i-Pr)2EtN. This compound is When applying the modi®ed Phillips's procedure8 to obtain considered a homologue of 1 0 -aza-C-uridine (Scheme 3). guanidine under re¯ux at 1008C for 10 h did not result in returned to the original Phillips's method. Thus the irradiation of b and a protons at H-2 0 and H-5 0 gave both 3±4% NOE in H-6 of 13 and 2% NOE in H-6 of 14.3-dimethyluracil (for 18). V. Full solubility of 9 is possible only in highly polar solvents (H2O. three days. treatment of (3R. V. Therefore.2) and C-6 (150.

4 Hz. 36. Irvine.6-di-O-isopropyl. 42. 3. 2H. Jˆ4. (3R.14. 1H. ol (11).25±7. 100%).00) for CDCl3. Calcd for C7H13N3O5´0. CA). 54. 3. 3-C-Azidomethyl-3-deoxy-1.9 (C-4). 3. 94%) MeOH/25% aq. Anal. Fmoc). Jˆ4. The reaction mixture was stirring for 30 min and cooling insoluble NaCl was ®ltered off.86 (m.40±3. 1H NMR (CD3OD) Merck and were visualized in an UV light (254 nm) and/or d 2. treated nations were performed using the 4.5.0 g.22 (m.6.1. 3.0 g.4 Hz.9 g. 7. 1. solvents were distilled before use.5 g. 75. Filichev.7 (CH [Fmoc]). 1H. and evaporated under reduced pressure to (100 mL) and washed with H2O (3£50 mL).1. H-5 0 (a)). n max (neat) 3417±2700 (br).45. mp 4. 128.6 mmol) in DMF stirring.4 Hz.43 (m. CHOH). 1H. 68. CD3OD (d : 49.85 mmol) was (100 mL) were heated with stirring at 958C for 3 h.6 (Fmoc). V.7 mmol) in H2O (10 mL) was added under vigorous (3. 4.06 (m. 8H. H. 3H.9166 V.5). 36.21 (d. 1H NMR (Me2SO-d6) d 2.11 (10% (d. H-5 0 (b)).4S)-4-[(1S. 1H.1.4. H-2 0 (a)).1. with a 5% solution of H2SO4 in methanol for sugar deriva. 4. 4£OH).0).06. N-Fmoc-(3R. 4.6 g.9. Pedersen / Tetrahedron 57 (2001) 9163±9168 with uracil and by alkylation with 6-chloromethyluracil in 4. 4. the was puri®ed on a silica gel column with CH2Cl2/MeOH resin was ®ltered off and the solvent was removed in vacuo. The residue was puri®ed by chromatography on a silica gel The excess of starting azasugar 12 was eluted with column with 20% MeOH in CH2Cl2 to give 8 (4.2R)-1.0). Jˆ4. 1H. 120. H. 64.2.6. Compound 614 (10. CH2N3).8 Hz.75H2O (232. 18. H-5). CH2OH). Sigma or Fluka.2.2. USA.1 (C-1).1.22 236±2398C (ethanol/few drops of H2O).5 MHz for 4. Me2SO-d6 (d : dissolved in a mixture of 10% aqueous NaHCO3 (25 mL) 39.2.3.7 Hz. 102.3 g. 1745. The solution was hydrogenated in an autoclave for 16 h at 200 psi.50±3. FAB-MS: m/z 400 (M1H)1.35 (m. 1H NMR steady-state NOE difference spectroscopy in dioxane (25 mL). 1H.2 (Fmoc).1. The silica gel (0. added. H-2. All ([CHOH]2CH2OH). 6. H-1).80 (m. 145. 1665 48. Internal standards used in 1H NMR spectra were pyl]pyrrolidin-3-ol (10). in 100 mL butan-1-ol and 3 mL HOAc) for azasugars and its 4£OH).2R)-1. 1H. General 80%) as a brown oil. The Rf and NMR data agreed with previous data. H-4). 3. 156.1.5 (C-3). 6.8 g.3. 49. H-3).1. H-2).8 (C-5).5 mmol) in EtOH (50 mL) a solution of NaIO4 (1. 72. The resulting solution was stirred at rt for 3 h. respectively. 10. the residue at 608C for 3. 68. 4H. 13C NMR (CD3OD) d derivatives. 1H.4S)-4-[(1S.4 mg.37 (10% MeOH/CH2Cl2).4.9 mmol) was were CDCl3 (d : 77. 2H.20 (m.8. 75.38. (0!25% MeOH) to afford the aza-C-nucleosides 13±15. 128.1 g) was added.92 (t. 5.2. 18.5 g. in 13C NMR hydroxypropyl]pyrrolidin-3-ol (9. 3-C-Azidomethyl-3-deoxy-d-allose (8.1. 26. After 30 min NaBH4 (139 mg.2R)-1. 70%) carried out with use of TLC plates 60 F254 purchased from as an oil: Rf 0.2.4 column chromatography was purchased from Merck.14 13±15 4. H-5).4 (C-4). The solution was ®ltered 4. 40% yield.23 (m. 1H.3 Microlab.0 (C-2). B. CH2OH). 2. Found: C.00. tives and/or with a ninhydrin spray reagent (0. E. . 42. 1H. 13.2 g. 1H. 4. 3.6 g. The combined organic layers were with toluene (30 mL).4 mmol) and 1. CD3OD. purchased from Aldrich.4 (CH2OH). 49.9 mmol) was a Bruker AC-250 spectrometer. Colorless powder. 54. After allowing the mixture to cool.7): C. Jˆ5. as a colourless oil: 1H NMR (Me2SO-d6) d 2.5.6 Hz. 2H. N. Rf and NMR data agreed with Na2SO4. The mixture was cooled at 0±58C and experiments were carried out on compounds 13 and 14 with 9-¯uorenylmethyl chloroformate (2. 4.0 g.41 4. Jˆ4. The combined organic layers were dried (Na2SO4) and The [M1H]1 and [M1Na]1 ions were peakmatched evaporated under diminished pressure to give an oil that using ions derived from the 2. To a cooled solution of compound 10 (1. 13C NMR (Me2SO-d6) d 44.1. 5. H-3). 1H.40 (m.4R)-4-(hydroxymethyl)pyrrolidin-3- 4.56 (m. MeOH/CH2Cl2).8.42 (m. 48. H-4). 1H. After cooling.2:5. A The azasugar 12 and 5-bromopyrimidines were mixed in the solution of azide 7 (7. of dimethylamine hydrochloride 7. General procedure for the synthesis of compounds 76%). 1H.5-dihydroxybenzoic acid was puri®ed by silica gel column chromatography with matrix. H-2 0 (b)). 9. N.45 (m.0 (CH2N3). 4.4R)-3-Hydroxy-4-(hydroxymethyl)pyrro- (t.31 (m.3-trihydroxypro- 13 C NMR.85 (br.98 (m.200) used for (C-2).3 g ninhydrin CH [Fmoc]).8 g. and 10% Pd/C (2.). Jˆ4. 63. lidin-3-ol (9). 126. 6.75 (d. 6H. Accurate ion mass determi.3-Trihydroxypropyl]pyrro- 65 and 50% yield. transform (FT) mass spectrometer (IonSpec. H-4 0 ). After added. NH3 (0!25%). 8. and the mother the solution was diluted with H2O (50 mL) and extracted liquor was evaporated to dryness in vacuo.11 using silica gel column chromatography with cyclohexane/ EtOAc (0!10% EtOAc) to afford compound 7 (6. The reagents used were 142. The mixture was fused in a preheated oil Amberlite IR-120(H1) (25 g) in H2O (100 mL) was heated bath at 145±1508C for 10 min. 5-[(3R. Microanalysis were performed by Atlantic 48. 3.8. 2. 3.1 (C-3). Experimental through Celitew and washed thoroughly with H2O and evaporated in vacuo giving the title compound 9 (1. 70.0 mmol).5 h.4S)-4-[(1S. idene-a-d d-allofuranose (7).2 Hz. 4H. 3-C-Azidomethyl-3-deoxy-d d-allofuranose (8). 75. 3.5 equiv. 3. The residue was dissolved in CH2Cl2 dried (Na2SO4).1.83 (m. 79. co-evaporated with CH2Cl2 (4£40 mL).6 cm21. 4. (dd. lidin-1-yl]uracil (13). freshly washed molar ratio 3:1. FAB-MS: m/z 242 [M1Na]1. 3.6 mmol) and NaN3 (2. [CH(OH)]2CH2OH. CH2 [Fmoc].88 (dd.78 (CHOH).28.3 mmol) was dissolved in 125 mL H2O.2. Rf and NMR data agreed with previous data. Thin layer chromatography (TLC) analyses were CH2Cl2/MeOH (0!10% MeOH) to afford 10 (1. 28.7 Tesla Ultima Fourier with H2O (50 mL) and extracted with EtOAc (3£75 mL). Jˆ5.3-Tri- TMS (d : 0. (3R. 71.063±0.5.11 NMR spectra were recorded on a Bruker AC-300 FT NMR spectrometer at 300 MHz for 1H NMR and at 75.0 g. 49.s. 3. N-Fmoc-(3R. Rf 0. dried over give pure 11 (1.1. and evaporated to give a syrup which was puri®ed previous data. Me2SO-d6.

10 (30% MeOH/ methyluracil-6-yl)pyrrolidine (18).9 (6-CH2N). 1H. (M1H)1 found 242. 73. Vol.4 mg. 41% yield. Khim.41 (20% MeOH/CH2Cl2). Ng. 1. Y.7 (C-4). 1998. 1H. 61. V. 31.8 (C-2).53 (m. 1H. E.7 Hz. 4.7 (C-2 0 ). 1214± (C-2 0 ).5 (C-4). Huang.68 (s. Acta Chem. 99. Jˆ8. 63. 3. B. Khalifa. Solvent was evaporated under diminished pressure.5 (C-5 0 ). Filichev.69 (dd.35 (s. 57. NH2).8 (6-CH2N). H-3 0 ). 3 0 -OH). 3. 2H. 2H. Jarvest.. Hansen. 907±915. 162. H-5 0 (a)). The residue was puri®ed on a silica gel column with 3863±3866.44 (s. H-5). 1H.. M. (C-5). Phillips.60 (br. E. 1H.. J. 2.s. H.35 (20% MeOH/CH2Cl2). 167. Chem. 142. M.s. R. 2£OH). A.1135. 1H NMR (CD3OD) d 2. H-3 0 ). using silica gel column chromatography with CH2Cl2/ V. 1H. Scand. Carbohydr..95 (s.2. 3 0 -OH). Chem. 6. 6. 128. 308. 1H. 74.5 (C-4 0 ). H-6). Bobek. 1H. 3H. 5-[(3R. The azasugar 12 (144 mg. H.3 Hz. (br. 2H. P. 55. H-2 0 ). 32. B. U. M. Jˆ5.1 (C-3 0 ). 1H.1139. A. 64. H-6).. cilylmethyl)pyrrolidine (17). Jˆ NMR (CDCl3) d 2. Med. HRMS: m/z (M1Na)1 found 250. 4. H-4 0 ). 51. J.64 (m.6 (C-5).1135. 4.4 (C-5 0 ). 2. Townsend. B.70 (m. Rf 0. Kim. (m.5 mL D. 2.s. 1H NMR prepared by the same methodology as described for the (Me2SO-d6) d 2.7 (C-4). 267± 4. Light-brown crystals. The Chemistry of C-Nucleosides.s. H-4 0 ).95 (br. CH2OH). V. NCH3).2. 7. The azasugar 12 (50 mg.1 (C-3 0 ).6 Hz. Zubarev and Mr B.0 References (CH2OH). M. 2H.0798. J.96 (dd. 1H. CH2OH. 57. Sùrensen. 6. 156. K.20±3. 4. mp 182±1838C (MeOH). 13C NMR (Me2SO. Ed. Koldobskii.95 (t.. 1±8.06 (m.48 mmol) 14. 5-CH2N). 56. H-5 0 2. (C-6). B. Bioorg.8 (C-2 0 ). p. 62.0 (C-2).2 (C-5).0. 13C NMR (Me2SO-d6) d 50. 333. 9.5. Jˆ5. 152. Pedersen. 1978. Pedersen. F.s. Lett. for 24 h. H-5 0 (b)).6. CH2N). Brandt.5 1222. D. 30.21 Yield 83%. J. L.8 (C-2 0 ). 1H. V.10 (m. P. 1H. Synthesis H-5 0 (a)). 1H. 2. 156. 9.7 (C-5 0 ).59 (s. 0. 71. 1H. Pedersen / Tetrahedron 57 (2001) 9163±9168 9167 1H. 2£OH). 3. Orgel.65 (m.83 (dd. Chem.8 Hz. (CD3OD) d 51. 3. 5.06 (t. HRMS: m/z 11.4R)-3-Hydroxy-4-hydroxymethyl-N-(6-ura. NCH3). NH)..8 161. Med.23 mmol) was dissolved in 1 mL (i-Pr)2EtN and 2. 421. 1989. 2H. K.3 227.90 d6) d 49.1 (C-4 0 ). Youn. H-5 0 (b)). Soc. Acton.3 Hz. K.0 Hz. 67.9 (C-2 0 ).8 (C-6). 2H. 150. 153. 5. G. Garegg.1 (C-2). 51. (NCH3).4-dioxane: Chem.4R)-3-Hydroxy-4-(hydroxymethyl)pyrro- lidin-1-yl]isocytosine (15). 56.38 (s. M.90 (t. N.. (C-6). 2. (3R. Y. (3R.90 (m. This compound was CH2Cl2). Jˆ2.06 (20% MeOH/ Acknowledgements CH2Cl2).. 8. n max (neat) 3366±2800. C12H20N3O4 requires 270. S. 12. H-2 0 . H-6). W. H.95 (t.90 (br.7 (C-2).7 (C-3 0 ).1 (C-5).20 (br. 4. Res. Chae. 2H.2. H-2 0 (b)). 25% NH3 /1. 4. 1H. 50%) as an oil: Rf 0.51 mmol) in 20 mL EtOH were re¯uxed 1757. 1H.6 (C-5 0 ). 3. Geterotsikl. (CD3OD) d 2. 1754± hyde (15. Chem. Carbohydr.. E. Lee. Soed. Rf 0.52 (m. E. J. D. Jˆ3.. 2H. 1H. 2£NH.2. 3. 13.. 2. J. H-5 0 (b)). E. 2. H-5 0 (a). 3. 13C NMR (Me2SO-d6) d 49.7 (C-5). 1H.1 (C-4 0 ). (CH2OH).7 (C-5). F. 150.. 1991. Goerner. 6-CH2N). 1H. Chem. Ryan. 4H. B-G. H-5 0 (a)). 1957.1. 3. Erussalimsky. 1H NMR (Me2SO-d6) d 2. 1965. 13C NMR (CDCl3) d 27. Carbohydr. Am.1123. n max (neat) 3352±2800.s. Davis.20 (m. MeOH and 6-uracilylmethylchloride (237 mg. (3R. CH2OH). Jaeger. 1H.4R)-3-Hydroxy-4-(hydroxymethyl)pyrro- lidin-1-yl]cytosine (14). 1998. Uh.5 Hz. J. CH2OH).43 (d. 3. V.3-dimethyluracil-6-ylmethyl 9.4R)-3-Hydroxy-4-hydroxymethyl-N-(1.05 1999. H-4 0 .2 (C-6).7 (C-2). 9.0 (CH2OH).9. Plenum: New York. MeOH (0!10% MeOH) to afford compound 17 (140 mg.4. H-5 0 (b)). 63.40 (m. H. 2.4R)-3-Hydroxy-4-hydroxymethyl-N-(5-ura. 1.. 1998. Jˆ Jˆ4. 52. 4.36 (m.50 (s. 2H. I. 6. 50/50). H-5 0 (b)). uracil (53 mg. 4H. 2H. 75. 4. Acc. 2£NH. 4.6 (CH2OH).7 Hz. 1H.7 (C-5 0 ). B.80 (br. OH). 62. H-4 0 ). 61. 74.2. H-2 0 high resolution mass spectra. A. L. W. 59.2 (C-2 0 ).s. 740±744. 32. 13C NMR (CD3OD) d 51. 8. S. I. 1H NMR 16. 1937±1943.1 Hz. 71. NH2).8. H-3 0 ). CH2OH). (b)).. C10H16N4O3 requires 242.. 101.8 H-3 0 ). 319± was added. 2H.3 Hz. E. 50. R. 125. H-2 0 (a)). Budnik for the 9.1123. V. Samuelsson. The 328.1150. G.1121. 1986. 1664 cm21. Light-brown crystals. S. J. HRMS: m/z (M1H)1 found (CH2OH). 4. N.0793. (67 mg. Titova.60 (t. B. 1951. 120. NH).6. 2. (a)).0 (C-4 0 ). HRMS: m/z (M1H)1 found 242.1 (C-4 0 ). 2H.92 (m. 166. Nikolaev. 1H. 7. H-2 0 chloride.8 (CH2OH). 4092. 1953. C10H16N4O3 requires 242. We thank Mr R. 1H. CH2OH). 1. L.. H-5).8 10. 5-[(3R. R. cilylmethyl)pyrrolidine (16). 9. 65%) as an oil: Rf 0. The mixture was stirred at rt for three days. H-3 0 ). 115±122. 0. Soc. 62. Watanabe. 117. 1H. CH2OH)..6-tri- mp 1388C (EtOH/5% CH2Cl2). C9H15N4O3 requires 227.20 (m.5 C9H13N3O4Na requires 250.0 Hz. Am. 6- 10. 3. H-4 0 . .10 (s. 4H. 164. 56. 1H. 1H (b)). 4.34 (s.95 (s. Harnden. 4. OH).7 (C-6).92 (dd.0 (5-CH2N). Bols. M. solvent was removed in vacuo and the residue was puri®ed 15.2. 3. F.43 mmol). 62. 153. 4.1139. 5. Poplavskii.. J. Chem.30 (s. C9H15N4O3 requires 227.58 (dd. 1979. 2£OH).9 (C-3 0 ). K. 1061±1062. 13C NMR 71. 1H. P.. Res.7 155. Jˆ5. 2. E. 3. 2H.47 mmol) and paraformalde. 73. 3.6 Hz.5 Hz.2 (C-3 0 ). K.65 (dd.1448.2 (C-4). 6. 270. 2. 53. Henry. 10. Chemistry of Nucleosides and Nucleotides. 1H.60 (br. 4H. 1H. 31. 518±525. B. H-3 0 ).3 (C-3 0 ). I. CH2OH.4. 131. Jˆ8.s.41 (s. Tetrahedron Lett.6 (C-5 0 ). 1647 cm21. E. Jˆ5.60 (m. 2. 51.9 (C-4). Rf 0.. A.30 (br. 156. Chem.. 1994. NH). H-6). 9. 4. 2. Filichev. 22. 0. 1086±1089. 1H. 151. 2H. Res. Bols.4 (C-6). Med. M. synthesis of 17 using 1. 7. HRMS: m/z (M1H)1 found 227. H-2 0 ). Res. 1991. 3. 3H.63 (aq. 1.0 (C-2). Org. CH2Cl2/MeOH (10!50% MeOH) to afford compound 16 6. 10. 1H. H-2 0 . Allen.1440. 1H. 4.88 (d. Biel. HRMS: m/z (M1H)1 found 270.4 (C-4 0 ). 41% yield.38 (m. 227. 287±290. Jˆ4.80 (s.2 (C-4).24 (m. A..10 (m.. M. M.1. Jˆ4. 2001. 56.3. 3. 110. 1H.3. Phillips. Biol.

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