3 346

4’-Substituted Nucleosides. 2. Synthesis of the Nucleoside Antibiotic Nucleocidin’ Ian D. Jenkins,2 Julien P. H. Verheyden, and John G . Moffatt* Contribution No. 121 from the Institute of Molecular Biology, Syntex Research, Palo Alto, California 94304. Received September 22, 1975

Abstract: The addition of iodine fluoride to N6,N6-dibenzoyl-9-(5-deoxy-2,3-O-isopropy~idene-~-D-er~f~ro-pent-4-enofuranosyl)adenine (9) leads to the formation of the corresponding 5’-deoxy-4’-fluoro-5’-iodo nucleosides with the P-D-ribofuranosyl (10) and a-L-lyxofuranosyl (11) configurations. Nucleophilic displacement of the 5’-iodo functions was very difficult but could be accomplished with azide ion giving the 5’-azido-4’-fluoro nucleosides (18, Ma). Photolysis of the azides followed by mild hydrolysis and borohydride reduction gave the corresponding 4’-flUOrO nucleosides (20, 2Oa) which were converted to their sulfamoyl esters (22, 22a) by way of intermediate 5’-O-tributyltin (21, 21a, R = S n B u j ) derivatives. Following removal of protecting groups the nucleoside antibiotic nucleocidin (1) and its cu-L-lyxofuranosyl epimer la were isolated. Several alternative methods for conversion of the 5’-iodo function in 10 and 11 to the hydroxy counterparts (20,ZOa) were investigated and this could be accomplished by oxidation with silver(I1) oxide followed by borohydride reduction. A number of consistent rules were established for assignment of configuration to 4’-fluoro-2’,3’-O-isopropylidenenucleosides from their N M R spectra. A number of other reactions of the 4’-fluoro-5’-iodo nucleosides are also described and some comments concerning the facile cleavage of the pyrimidine ring in N6-acetyl-N3,5’-cycloadenosinederivatives are presented.

The microorganism Streptomyces calvus, isolated from a n Indian soil sample, was shown in 1957 to elaborate a n antibiotic substance related to adenosine and referred to as nu~ l e o c i d i nThis . ~ substance was shown to exhibit a rather broad antibacterial spectrum and to be particularly active against trypanosome^.^ Its practical use as either a n antibiotic or antitrypanosomal agent is, however, seriously limited by its toxicity, the LD50 of nucleocidin in mice being 0.2 mg/kg via ~ intramuscular or intraperitoneal a d m i n i ~ t r a t i o n .Nucleocidin has been shown to be a highly potent inhibitor of protein biosynthesis, but its precise mechanism of action has not been clearly defined.5 Several partial and complete structures were proposed for nucleocidin,6 but not until 1969 was it realized that the empirical formula properly contained fluorine.’ With this realization it was possible, by use of N M R and mass spectrometry, t o conclude that nucleocidin was correctly represented as 4’fluoro-5’-O-sulfamoyladenosine (1) or a n isomer thereof. NH, I

1

Subsequent work by Shuman et a1.8 supported the P-D configuration for the pentose moiety since upon heating in dimethylformamide nucleocidin was converted to a n N3,5’anhydronucleoside. Assignment of the D-rib0 configuration remained only inferential but is now confirmed by the total synthesis reported in this paper. T h e structure 1 for nucleocidin is unique in several ways. Thus, it is, to the best of our knowledge, the first natural product to contain either a fluoro carbohydrate derivative or a n unsubstituted sulfamoyl group. It also appears to be the first example of a furanose sugar bearing a functional substituent a t C4. This combination of structural features made the total synthesis of nucleocidin an attractive synthetic challenge, and its successful accomplishment is reported in this paper. A preliminary account of this work has appeared p r e v i o ~ s l y . ~ The major synthetic problem in a synthesis of 1 was certainly

Journal of the American Chemical Society

the stereospecific introduction of the 4’-flUOrO function. Previous work in this laboratory has led to the development of general methods for the synthesis of 4’,5’-unsaturated nucleosides in both the pyrimidine” and purine series” and related studies have been undertaken by others.I2 An attractive route for introduction of the 4’-fluOro moiety appeared to be the addition of a suitable fluorine-containing pseudohalogen such as iodine fluoride to a suitably protected 4’,5’-unsaturated adenosine derivative. Iodine fluoride, which can be isolated from the reaction of iodine and fluorineI3 or can be generated in situ from silver fluoride and iodine, is known to add to olef i n and ~ ~to vinyl ~ ~ ethers such as g 1 y ~ a l s . There I ~ ~ was, however, some discouraging precedent for this type of reaction since McCarthy et a1.’2b had attempted the reaction of the olefin 2a with bromine in methanol. T h e product of this reaction proved to be the N3,4‘-anhydronucleoside (4a), which undoubtedly arose via attack of N3 of the adenine ring upon Cq, of the initially formed 4’,5’-bromoniurn ion 3a or its oxonium ion counterpart 3b. The above formation of 4a bears a close resemblance to the well-known, and frequently spontaneous, intramolecular displacement of electronegative C y substituents leading to N3,5’-cycloadenosines.‘S Previous work by Jahn16 has shown, however, that acylation of the 6-amino function in adenosine derivatives reduces the electronegativity of N3 to the extent that the formation of N3,5’-cyclonucleosides is greatly suppressed. W e have also made considerable use of this observation during various studies on the preparation of 5’-halogenated purine nucleosides,” and it seemed worthwhile to first of all examine a model reaction similar to that used by McCarthy et al. I 2b Accordingly, 2’,3’-O-isopropylideneadenosinewas converted into its N6-benzoyl derivative 6aI7 using a modification of the method of Chlddek and Smrt.Is Without purification, crude 6a (80% yield and 95% purity) was converted essentially quantitatively into its 5’-0-methanesulfonyl derivative 6b. While 6b could be completely purified by preparative T L C a s judged by T L C and N M R analysis, it was not obtained in crystalline form. Treatment of the crude product with a n excess of potassium tert-butoxide in tetrahydrofuran a t -50° readily led to the isolation of crystalline N6-benzoyl-9-( 5-deoxy-2,3- O-isopropylidene-P-D-erythro-pent-4enofuranosy1)adenine (2b) in a n overall yield of 58% from

2’,3’-O-isopropylideneadenosine. A methanolic solution of the olefin 2b was then allowed to react a t room temperature with a slight excess of bromine

/ 98:11 / M a y 26, 1976

3347

HNR

I

0 x 0 2a,

R

b, R

=H = Bz

r

1

3a

3b NHBz

“R

4a,

R =H

5

b, R = B z giving a mixture of the desired 5’-bromo-4’-methoxy nucleosides 7 and 8 in 46% yield and in a ratio of 2:l as judged by N M R (see later). By careful preparative T L C 7 and 8 could be separated and isolated as homogeneous foams. Assignments of stereochemistry to 7 and 8 could be made based upon their ~ in one isomer were magN M R spectra. T h u s the C Sprotons netically equivalent and appeared a s a 2-proton singlet, while in t h e other isomer these protons were nonequivalent and appeared as a pair of doublets. Since a cis arrangement of the Cs+bromomethyl and the 2’,3’-O-isopropylidene groups would be expected to inhibit free rotation of the C 4 4 5 ’ bond, the NHBz

I

N N -

2b

---+

Br,

MeOH

0 x 0 6a, R = H b. R = M s

HNBz

HNBz

I

I

7

8

isomer showing nonequivalent Cgf protons is assigned the CYL-lyxofuranose structure 8. As a further confirmation, a cis relationship between the C~J-methylene group and C3.H in the O-D-ribo isomer 7 was demonstrated by a substantial nuclear Overhauser e n h a n ~ e m e n t of ’ ~ the integrated intensity of the C3iH signal upon irradiation of C5.H. A s will be seen later in this paper, the chemical shift difference between the isopropylidenemethyl signals (A6 27 H z for 7 and 18 H z for 8) also confirms the assigned configurations. T h e presence of the N6-benzoyl function thus clearly had the desired effect in inhibiting the formation of a n N3,4’-anhydronucleoside. In addition to 7 and 8 the above reaction gave 28% of a n unresolved mixture of 5’-bromo-4’-methoxy nucleosides lacking the 2’,3‘-O-isopropylidene function and only 15% of a co’mpound considered on the basis of elemental analysis and spectral measurements to be the ring-opened N3,4’-anhydronucleoside 5 rather than the expected 4b. Recently Sasaki et al.20 have briefly reported a compound considered to be 5 from the reaction of 2b with N-bromosuccinimide and methanol. T h e N M R and uv spectra of 5 described by these workers a r e somewhat different from those we report, but since this compound is peripheral to our central interest we have not repeated the preparation to clarify the situation. Some further comments concerning the spontaneous ring opening of N-acylated cycloadenosine derivatives will be found later in this paper. T h e relatively successful reaction of 2b with bromine and methanol encouraged us to examine the addition of iodine fluoride. In fact, the reaction of 2b with a mixture of silver fluoride and iodine14 in tetrahydrofuran did lead to a mixture of fluoroiodo adducts. These compounds, however, had very similar T L C mobilities and could not be efficiently separated. W e then considered the possibility that addition of a second benzoyl group to the adenine ring of 2b might not only further stabilize the molecule against anhydronucleoside formation but also facilitate separation of the isomeric adducts. Accordingly, 2b was allowed to react in pyridine with benzoyl chloride giving a n almost quantitative yield of N6,N6-diben-

zoyl-9-(5-deoxy-2,3-O-isopropylidene-~-~-erythro-pent-4enofuranosy1)adenine (9). Such dibenzoyladenine nucleosides have frequently been assigned the N 6 ,1-dibenzoyl sructure, but recent studies by Anzai and Matsui,*la subsequently confirmed by Lyon and Reese,*Ib have shown that the N6,N6-dibenzoyl structure is correct. As part of a separate problem to be described later we have also confirmed this conclusion via an examination of the I3C N M R spectra of a number of dibenzoyladenine nucleosides. Complete debenzoylation of 9 with methanolic ammonia gave the known crystalline 9-( 5-deoxy-2,3-0-isopropylidene-@-~-erythropent-4-enofuranosyl)adenine (2a) in 93% yield. The reaction of 9 with iodine fluoride generated in situ from silver fluoride and iodine has been examined under a variety of conditions and leads, in most cases, to the formation of a mixture of two isomeric 4’-fluoro-5’-deoxy-5’-iodo nucleosides (10 and 11) in 60-9oOh yield. These two products can be cleanly separated and isolated in analytically pure form by careful preparative T L C using a 40-cm development. On a preparative scale it is more convenient to use chromatography on a column of silicic acid, which effects a partial separation giving pure samples of both 10 and 11 together with some mixed fractions. Rechromatography of the latter permits further resolution and isolation of pure materials. From the N M R spectra of 10 and 11 it was clear that, as expected, fluorine was introduced a t Cd, of the sugar moiety since both C3lH and C5fH2showed vicinal coupling to fluorine. This point was confirmed by catalytic hydrogenolysis of the 0-D-rib0 isomer 10 (see below) which afforded the corresponding 5’-deoxy-4’-fluoro nucleoside 12, the N M R spectrum of which showed the 5’-methyl group a s a 3-proton doublet strongly coupled to the C4t-fluorine ( J ~ J . F = 17 H z ) .

Jenkins, Verheyden, Moffatt

/ Nucleoside Antibiotic Nucleocidin

3348 NBz,

generally indicative of a physical proximity of the nuclei inv 0 1 v e d ~and, ~ since it is not observed in the P-D-ribo isomers N+N (e.g., l o ) , adds further support to the above assignments. It should be noted that the Cy protons in most of the 4’-fluoro I nucleosides described in this paper give NMR signals that are frequently deceptively simple and not readily amenable to first-order analysis. These ABX patterns have been subjected to computer analysis using established procedures26 in order to provide the parameters given in Tables I and II.*’ 0 0 oxo Chemical support for the above configurational assignments X was also sought via formation of an N3,5’-anhydronucleoside. 9 10 Thus, small samples of 1 0 and 11 were first debenzoylated with Bz, NHi methanolic ammonia and the resulting nucleosides were directly heated in dimethylformamide. While most 2’,3’-@isopropylideneadenosine derivatives bearing electronegative ~ N3,5’-anhydronucleosideson gentle substituents a t C S form warming,’s the presence of the 4’-fluor0 function enormously inhibits this reaction. It was necessary to heat the 0-D-rib0 isomer 1 0 a t 140’ for 16 h in order to get roughly 20% conversion to a n ionic substance (13a) with an ultraviolet spectrum and electrophoretic mobility essentially identical with those of a n authentic sample of 2’,3’-0-isopropylidene-N3,5’-cycloadenosine iodide28 (13b). Even under these conditions 11, 12 13a, X = F 11 in which CS,and the purine ring a r e trans disposed, failed to b,X=H give any ionic material similar to 13. The assignments of configuration for 1 0 and 11 were based The relative proportions of 1 0 and 11 from the condensation primarily upon N M R spectroscopy and were supported by of 9 with iodine fluoride varied markedly with the precise rechemical evidence. T h e N M R spectra of 10 and 11 a r e sumaction conditions. Under many conditions (e.g., slow addition marized in Tables I and 11. T h e most significant differences of a solution of 5 equiv of iodine in tetrahydrofuran to a sus~ in the in these spectra were the appearance of the C Iproton pension of 6 equiv of silver fluoride and 9 in methylene chloride less polar isomer as a doublet of doublets showing Jlt.2, = 0.5 a t room temperature) the predominant product is the undesired H z and five-bond long-range coupling to fluorine ( J I , , F= 2.5 a - ~ - l y x oisomer 11. In the above experiment the isolated yields Hz). T h e more polar isomer showed only a small vicinal couof 1 0 and 11 were 8 and 71%, respectively. Many variations N 1 Hz) and no coupling to fluorine. Previous work pling of solvent and condition were explored, ultimately showing that 10 could be obtained a s the major product by the gradual adby Hall et aLZ2on glycofuranosyl fluorides has shown that five-bond H - F coupling (JF’.H4 = 6-7 Hz) occurs in such dition of solid iodine to a relatively dilute (0.07 M ) solution of compounds when the C I - F and C4H functions a r e trans dis9 in methylene chloride a t room temperature in the presence posed, whereas the corresponding cis isomers show much of freshly powdered silver fluoride (see the Experimental smaller values ( 1 -2 Hz). Based upon the above the less polar Section). T h e optimal conditions in terms of both yield and ratio of products appeared to involve gradual addition of a isomer, which shows Hl,-F coupling, may be assigned the N6-dibenzoyl-9-( 5-deoxy-4-fluoro-5-iodo-2,3-0-isopropyli-solution of iodine in acetonitrile to a dilute solution of 9 in dene-a-L-1yxofuranosyl)adeninestructure 11 while the more acetonitrile in the presence of silver fluoride a t -40’. Under polar product is the desired 0-D-ribofuranosyl isomer 10. it a variety of such conditions mixtures of 1 0 and 1 1 were obtained in combined yields of 7 5 4 5 % and in ratios of 1: 1 to 2: 1, may be noted that, as expected, H1f-F coupling is not observed in the spectrum of nucleocidin itself‘ and the somewhat smaller The iodine can be replaced by N-iodosuccinimide or the silver fluoride by mercuric fluoride without greatly changing the coupling observed for 1 1 relative to those in acylated glycoproduct distribution. The choice of solvent does not seem to be furanosyl fluorides** is probably a consequence of the rather rigid conformation induced by the 2’,3’-O-isopropylidene critical, roughly comparable results arising from reaction in function. Indeed, it can be seen from Tables I and IJ that the methylene chloride, tetrahydrofuran, acetonitrile, and benzene. T h e use of nitromethane seems to give a favorable ratio (2: 1 ) spectrum of the a-L-lyxofuranosyl isomer of nucleocidin, deof 1 0 and 11, but the overall yield is lower (-50%) and a yelrived from 11, shows a value of J11.F = 7 Hz. Similar results low, crystalline by-product is formed that appears to be a have been obtained with some 2’,3’-di-O-acyl analogues of 1 0 mixture of two isomeric nucleosides by N M R . W e have not, and 11 for which values of J ~ , , = F 0 and 5-6 Hz, respectively, a s yet, been able to characterize this substance. have been observed.23 Since under all conditions investigated a considerable Further support for these assignments is to be found in a n amount of the IF adduct with the undesired a-L-lyxo configexamination of the N M R signals for CyH in 1 0 and 11. It has uration ( 1 1 ) was formed, it was of interest to attempt the previously been ~ h o w n ~ *in~ -studies * ~ on a variety of fluoequilibration of this product to its more interesting 0-D-rib0 rinated furanose sugars that trans-vicinal H - F couplings a r e isomer. It was hoped that in the presence of boron trifluoride quite large (JH,F = 20-30 Hz) while cis-vicinal couplings a r e = 4-10 Hz). In agreement with this the 11 would be in equilibrium with the oxonium species 1 4 which much smaller (JH.F P-D-ribo isomer 1 0 showed J 3 ‘ , F = 1 1.5 Hz while the cis discould add fluoride in regenerating a mixture of 1 0 and 11. In posed cl-L-lyxo isomer 11 had a considerably smaller J ~ J ,ofF fact, treatment of an enriched sampleof 11 (11:lO = 3 : l ) with boron trifluoride etherate followed by addition of finely divided 5.5 Hz. Once again. the absolute values of these couplings a r e sodium fluoride (silver fluoride and tetramethylammonium smaller than those in more flexible acylated sugars due to the fluoride seemed to behave similarly) led to the formation of isopropylidene group but the relative magnitudes strongly confirm the assigned configurations. Finally, the a - ~ - l y x o a major product with the same T L C mobility as 10. Isolation isomer 11 and other related compounds in this paper show a of this material, however, readily gave a crystalline compound identified as N6-dibenzoyl-9-(5-deoxy-.5-iodo-2,3-0-isoprosmall ( J = 0.5- 1.5 Hz) but clearly apparent “through space” pylidene-~-~-erythro-pent-4-enofuranosyl)adenine (15) in coupling of C4fFto CgH of the purine ring. Such a n effect is NBz,

I

I

4,’ ,P

=.p& -j

-

+

(J11.21

Journal of the American Chemical Society

1 98:l I /

May 26, 1976

3349 Table I.

100-MHz NMR Chemical Shifts

Compd

Solvent'

P P

C1.H

C2.H

C3.H

C4.H

CSH~

5.25 (dd) 5.67 (ddd) 5.50 (d) 5.32 (d)

5.56 (dd) -4.90 (m) 5.86 (br d) 5.55 (dd)

4.96 (dd) 4.51 (d) 4.53 (d)

C5,Hb

5.03 (d) 5.21 (dd) 4.66 (br d) 4.67 (dd)

CzH, CsH

Other

8.49, 8.50 (s) 8.58, 8.60 (s) 8.32, 8.56 (s) 8.05, 8.77 (s)

8.23 (br s, 2, "2) 8.20 (br s, 2, "2) 1.42, 1.56 (s, 3, CMe2) 1.42, 1.57 (s, 3, CMe2), 7.53, 8.02 (m, 5, Ar), 9.15 (s, N H ) 1.27, 1 S O (s, 3, CMe2), 7.4 (m,4, Ar and C2H), 8.0 (m, 3, Ar and NCHO) 1.36, 1.62 (s, 3, CMez), 7.5, 7.99 (m. 5, Ar)

1 la 2a 2b

C

6.93 (d) 7.03 (dd) 6.71 (s) 6.31 (s)

5

C

5.38 (s)

4.78 (d)

4.93 (d)

3.99 (d)

4.18 (d)

7.4

6a

C

5.96 (d)

5.20 (dd) 5.04 (dd) 4.49 (m)

3.74 (dd)

3.96 (dd)

8.09, 8.70

6b

C

6.16(d)

5.44(dd) 5.10(dd) 4.45(m)

4.45(m)

4.45(m)

8.11,8.74

P

3.64 (s)

5.31 ( b r s ) 5.31 (s)

8.1 I , 8.76 (s)

7

C

6.21 (d)

8

C

6.51 (br s) 5.55 (br d) 4.92 (d)

9

C

6.27 (br s) 5.26 (d)

10

C

6.36 (br s) 5.27 (dd) 5.41 (dd)

3.50 (ABX)* 3.54 (ABX)

8.15. 8.67 (s)

11

C

6.48 (dd) 5.58 (dd) 5.08 (dd)

3.54 (ABX)

12

C

6.30 (d)

5.30 (dd) 5.11 (dd)

8.24 (br s), 8.70 (s) 8.13, 8.69 (s)

15

C

6.38 (s)

5.36 (d)

16

C

17

5.49 (br d)

3.52 (d)

3.71 (d)

8.13, 8.79 (s)

4.49 (d)

4.63 (d)

8.05, 8.62 (s)

3.57 (ABX)

1.65 (d) 5.45 (s)

8.10, 8.60 (s)

6.37 (br s) 5.32 (br d ) 5.54 (dd)

5.32 (d)

8.13, 8.63 (s)

C

6.42 (dd) 5.64 (dd) 5.22 (dd)

5.33 (d)

18

C

6.38 (br s) 5.34 (dd) 5.55 (dd)

8.23 (br s), 8.66 (s) 7.93. 8.41 (s)

18a

P

6.93 (dd) 6.04 (br d) 5.56 (dd)

20

P

6.90 (br s) 5.53 (dd) 5.90 (dd)

20a

P

6.91 (dd) 5.93 (dd) 5.57 (dd)

4.28 (ABX)

4.38 (ABX)

21 ( R = Ac) 21a

C

6.28 (br s) 5.28 (dd) 5.45 (dd)

4.26 (dd)

4.45 (dd)

C

6.51 (dd) 5.61 (dd) 5.13 (dd)

4.50 (ABX)

4.55 (ABX)

22

P

6.86 (br s) 5.40 (dd) 5.92 (dd)

4.83 (dd)

4.96 (dd)

22a

P

6.80 (d)

5.49 (dd)

4.88 (ABX)

4.92 (ABX)

23a

P

6.90 (dd) 5.93 (dd) 5.48 (dd)

3.33 (ABX)

3.36 (ABX)

23b

C

6.47 (br d) 5.52 (d)

5.09 (dd)

3.81 (ABX)

3.86 (ABX)

24

D

6.92 (s)

4.85 (d)

5.30 (d)

5.27 (br s) 5.05 (dd)

5.66 (dd)

8.17 ( b r s ) , 8.66 (s) 9.02, 9.40 (s)

25

C

5.83 (s)

4.70 (m)

4.70 (m)

4.70 (m)

4.93 (dd)

7.50 (s)

5.59 (d)

3.61 (d) 3.74 (ABX)

3.79 (ABX)

8.52 (d), 8.80 (s)

4.19 (d)

(R = Ac) 5.95 (d)

8.46, 8.56 (s) 8.55 (d), 8.76 (s) 7.81, 8.30 (s) 8.03 (d), 8.42 (s) 8.47, 8.52 (s) 8.38 (d), 8.64 (s) 8.54 (d), 8.80 (s)

Solvents are: C, CDCI3; P, pyridine-ds; D, DMF-d7. by computer a n a l y ~ i s . ~ ~ - ~ ~ (I

3.04 (dd)

1.37,1.60(s,3,CMe2),

2.89 (s. 3. SO7Me). 7.5, 7.98 (m, i,A;) 1.39, 1.66 (s, 3, CMe2), 3.47 (s, 3, OMe), 7.5 and 8.0 (m, 5, Ar) 1.42, 1.60 (s, 3, CMe2), 2.93 (s, 3, OMe), 7.5 and 8.0 (m, 5, Ar) 1.42, 1.56 (s, 3, CMez), 7.3, 7.8 (m, IO, Ar) 1.38, 1.63 (s, 3, CMe2). 7.4, 7.85 (m, 100 Ar) 1.40. 1.58 (s, 3, CMel), 7.4, 7.85 (m, I O , Ar) 1.37, 1.62 (s, 3, CMe2). 7.4, 7.85 (m, IO, Ar) 1.42, 1.55 (s, 3, CMel), 7.4, 7.85 (m,IO, Ar) 1.40, 1.66 (s, 3, CMe2). 7.4, 7.8 (m,IO, Ar) 1.40, 1.59 (s, 3, CMe2). 7.35, 7.8 (m,IO, Ar) 1.41, 1.65 (s, 3, CMez), 6.24 (br s, 2, "2) 1.36. 1.53 (s, 3, CMeZ), 8.44 (br s, 2, "2) 1.43, 1.73 (s, 3, CMe2), 8.29 (br s, 2, "2) 1.35. 1.52 (s, 3, CMe2), 8.34 (br s, 2, "2) 1.39, 1.63 (s, 3, CMe2), 5.89 (br s, 2, "2) 1.41. 1.58 (s, 3, CMe2), 6.61 (br s, 2, "2) 1.38. 1.67 (s, 3, CMer), 8.39 (br s, 4, "2) 1.31. 1.47 (s, 3, CMe2), 8.31 (br s, 2, "2) I .37. 1.54 (s, 3, CMe2), 8.40 (br s, 2, "2) 1.40, 1.58 (s, 3,CMe2), 7.3, 7.8 (m,IO, Ar) 1.25. 1.49 (s, 3, CMeZ), 2.50 (s, 3, NAc), 8.02 (s, 1, N H ) I .28, 1.50 (s, 3, CMel), 2.49 (s,3,NAc).8.48 (s, I , NCHO)

The ABX patterns for the Cy protons of 10, 11, Ma, 20a-23a, and 23b were derived

47% yield. T h e elemental analysis of 15 clearly indicated the loss of the elements of hydrogen fluoride from 11 and the N M R spectrum showed the absence of a 4' proton and the presence of only a single vinylic 5' proton a t 5.45 ppm. There was no indication of allylic (3',5') coupling, suggesting that these protons a r e trans oriented relative to the double bond.

It is known that cisoid allylic couplings are generally somewhat larger than t r a n ~ o i d and , ~ ~it has been observed that only one of the 5' protons in nucleosides such as 2a, 2b, 9, or their pyrimidine counterpartsI0 is allylically coupled to C3.H (J3',5'b = 1 Hz). The presence of the double bond is also supported by a positive spray test with dilute potassium permanganate on

Jenkins, Verheyden, Moffatt

/

Nucleoside Antibiotic Nucleocidin

3350 Table 11.

First-Order Coupling Constants, Hz“

Compd

JI,,~,

1 la 2a

2 7

JI,.F

J2‘.3’

0

6

17.5

7

b

0

7 6

2b

0

5 6a 6b 7 8 9

0

4 2 0.5

J4’.5‘a

J31.4,

8.5 14

J4‘.5%

J5’a,5’b

8.5 20

10

0 J2’,4‘

=3

-

2

J3’.5’a = 0 , J3f,j’b

6

2

J3’,5’a = J3’.5’b

5 6 6

13 12.5

1.5 3

2.5

2

b

b

= 1

b

5.5 6

12 3

J3,,5,a =

0,

J3’,s’bN

10 11 12 15 16 17 18 1Sa 20 2Oa 21 ( R = A c ) 21a (R = Ac) 22 22a 23a 23b 24 25

-I

6.5 5.5 6.5 6

11.5 5.5 12.5

18.24 14.54 17

0

6

2.5

5.5

0

6 6

12.5 4.5 13 7 12 6 12 5.5 12 5.5 6 5.5

17.5 26.5 13.5 19.56 9.5 10.63

0

0.5

2.5

1

0

0 -1 1

-I

0.5

2.5 2.5

6.5 6

0

6

2

5.5 6 5.5

0

-1 1

-1 1