J. Org. Chem., Vol. 4 4 , No. 1, 1979 9
Streptozocin (3) H. Hart and B. Huge, Tetrahedron Lett., 3143 (1977). (4)H. Hart, J. B-C. Jiang, aind R. Gupta, Tetrahedron Lett., 4639 (1975). (5) W. Mosby, J. Am. Chem. SOC.,74,2564 (1952): J. Colonge and L. Pichat, C. R. Hebd. Seances Acad. Sci., 226, 673 (1948). (6) J. B-C. Jiang, Ph.D. Thesis, Michigan State University, 1975. (7) This reaction was developed by Mr. David Makowski. (8) E. Wolthuis. J. Org. Chom., 26,2215 (1961). (9) We do not imply a bisaryne intermediate; the additions might proceed through such an intermediate or they might proceed stepwise. (IO) G. Wittig and H. Harie, ,Justus Liebigs Ann. Chem., 623, 17 (1959), con-
sidered the possibility of generating bisarynes From 2,6-difluoro-3.5dibromo-pxylene, either magnesium in THF or butyllithium in ether, and furan they obtained mono- and bisadducts, the latter in only 5-15% yield. (1 1) G. W. Gribble, R. W. Allen, P. S. Anderson, M. E. Christy, and C. D. Colton, Tetrahedron Lett., 3673 (1976). (12) The last two steps in the synthesis were carried out in a nitrogen atmosphere. (13) E. Wolthuis, W. Cady, R. Roon, andB. Weidenaar, J. Org. Chem., 31, 2009 ( 1966).
Streptozocin: Structure and Chemistry' Paul F. Wiley," Ross R. Herr, Heinz K. Jahnke, Constance G. Chidester, Stephen A. Mizsak, L. Bayard Spaulding, and Alexander D. Argoudelis* Research Laboratories, T h e Upjohn Company, Kalamazoo, Michigan 49001 Received June 2,1978
The struci u r e of streptozocin has been shown t o b e t h a t represented by 1. Its degradation by a variety of reagents w i t h loss of the nitroso group a n d formation o f new r i n g systems is discussed. and the structures of the products are reported.
Streptozocin? (l),an antibiotic produced by Streptomyces achromogenus sub. streptototicus, is a broad spectrum antibacterial agent.S4 [t is also an antitumor agent being used clinically for malignant islet cell cancers of the p a n ~ r e a sIn .~ this report we wish ih present evidence establishing the structure of streptozocin to be 1 and to discuss its chemistry. Streptozocin (1) has a molecular formula of C~H15N307~ established by analysii3 and molecular weight determination. Its ultraviolet and infrared spectra have been reported.6 The two maxima in the ultraviolet [228 nm ( t 6360) and 380 nm ( 6 136)] are consistent with the presence of an N-nitroso group.7 The infrared spectrum has bands suggesting OH/" and a carbonyl group.6 A potentiometric titration showed the absence of titratable groups. The lH NMR spectrum of 1 was not well-resolved and could not be completely assigned, but a singlet a t 6 3.15 representing 3 H indicated CH3N. No CH3C groups were present. A 13C NMR spectrum (DzO a t pH 4.3) had chemical shifts of 6 156.7 and 28.4, confirming the presence of a carbonyl and a methyl group. The remaining signals were virtually identical with those of a mixture of CY- and 02-acetamido-2-deoxy-~-glucopyranose,8 suggesting that Dglucosamine in the pyranose form is the nucleus of 1. 1 is readily converted to a tetraacetyl derivative (2) by the acetic anhydride-pyridine procedure (Scheme I). The l H NMR spectrum of 2 showed the presence of four CH3CO groups and the CH3N group. A multiplet a t 6 3.93-4.60 arose from 3 H on carbons substituted by oxygen or nitrogen. Two triplets ( J = 10 Hz in each case) at 6 5.11 and 5.67, each representing one H, are consistent with the resonances of protons on C-3 and C-4 on a glucosamine skeleton. Treatment of 1 with 2 N NaOH solution gave diazomethane determined by conversion of p -nitrobenzoic acid to its methyl ester6 and an amorphous solid (3)having the molecular formula C;.H11N06. Acet,ylation of 3 gave a crystalline solid (4).Ia Hydrolysis of both 3 and 4 with 2 N HCl gave glucosamine hydrochloride, identified by comparison with an authentic sample, and carbon dioxide. The spectral data and formation of diazomethane after treatment with alkali establish the presence in 1 of the group CH,N(NO)C(=O)-. The isolation of glucosamine accounts for the remainder of the molecule. Since there is no basic group in 1, the amine in glucosamine must have been the site of attachment of the carbonyl group 0022-32S3/79/1944-0009$01.00/0
in the above moiety. The conversion of 1 to a tetraacetyl derivative and its NMR spectra establish that there has been no rearrangement of the six-carbon fragment on hydrolysis. The 13C NMR spectrum of I and mutarotation undergone by 1 from widely varying original values of different lots to a constant value of +39O in water indicate the presence of CY and @ forms. In fact, methyl glycosides of the two isomers have been preparedagThus, the structure of streptozocin must be as indicated in formula 1.Acetylation gave the 0 isomer (2) as indicated by the coupling constant of 8.5 Hz for the HI,* coupling. The structure of 1 was confirmed by synthesis in the original work,la and subsequently two improved syntheses have been reported.loJ1 A number of degradative conversions of 1 by various reagents have been brought about, and the compound itself in various solvents such as water, ethanol, and MezSO undergoes spontaneous decomposition. Loss of the nitroso group occurs in all reactions of this type. Some of the degradations have already been reported,lJ2 but they will be discussed further in the present publication. The formation of 3 from 1 as a result of base treatment has been reported,la and a structure (12)was proposed for it with the assumption that the pyranose ring form was retained. The tetraacetyl derivative of 3 was thought to have structure 13 largely because of a 1790-cm-l band in its infrared spectrum and one CH3C signal in the lH NMR spectrum of 4 differing markedly from the other three. The infrared spectrum of 4 has three bands in the carbonyl region at 1790, 1735, and 1705 cm-l, suggesting acetate (1735 cm-l), a carbonyl similar to the one in 3 (1705 cm-l), and a new group giving the high wavenumber band. The l H NMR spectrum has signals at 6 1.95, 2.01, and 2.03 which must be from acetyl CH3C groups attached to oxygen, while a fourth resonance at 6 2.42 suggests some other type of acetyl group. The other chemical shifts were as expected for a sugar derivative except that the H2,3 coupling was zero, perhaps indicating a furanose ring. The 13C NMR spectrum was reasonable for both 4 and 13.As a result of inconsistencies in the data derived from the alkaline degradation product of 1 and its acetyl derivative, an X-ray crystallographic study was done on the acetyl derivative. Its structure was established as that represented by 4, and, as acetylation should not cause rearrangement of the ring structure, the structure of the initial degradation product
0 1979 American Chemical Society
10 J . Org. Chem., Vol. 44, No. 1, 1979
Wiley et al. Scheme I
H > O Cj :{
CH,COO NHCNCH
I
NO 2 CH OH I
PYI
't
CH ,CON
C H O H ''
I
N() 1
I
9
10
OCOCH
v
A H 11
Numbering i n ;11! c o m p o u n d s i < the same as t h a t of the corresponding atoms in 1.2
CH-OCOCH
J . Org. Chem., Vol. 44, No. I , 1979 11
Streptozocin
Table I. Final Atomic Parameters (XlO4) and Standard Deviations (in Parentheses)O x, O(1) C(2) C(3) C(4) NI5) C(6) O(6) O(7) C(8) C(9) CI10) O(11) C(l2) O( 12 I C(l2M) O(13) C(14) O(141 C(14M) 0115) C(16) D(16) C(l6M) C(17) O(17) C(17M) a
12) 873 1 3 ) 1083 1 3 ) 914 ( 3 ) 2203 ( 3 ) 2518 ( 3 ) 3530 ( 2 ) 1470 ( 2 ) 345 1 3 ) 175 ( 3 ) -178 1 3 ) 998 ( 2 ) 847 ( 3 ) -234 ( 2 ) 2169 141 1172 12) 706 ( 3 ) -473 ( 2 ) 184b 1 3 ) 5 (21 343 1 3 ) 1484 12) -873 14) 2940 1 3 ) 250R 12) 4193 ( 3 ) -4
2
Y
4207 3668 2276 2323 2331 3535 3885 4338 3649 3897 5285 6063 6999 7288 7591 3472 2836 2580 2507 1495 612 466 -94 1194 272 1179
(1) (2) (2) (2) (2) 13)
(2) 12) (2) (21 (21 12) (2) (2) (3)
(2) (2) 12) 13)
(1) (2) 12) (3)
(3) (2) (4)
1568 959 1200 2131 2552 2846 3191 2674 2304 140 1 187 739 1032 93 1 -446 -1103 -1203 -1655 888 320 73 57 2613 2255 3129
811
11) 11) 1)
80 55 47 47 65 100 126 117 81 58 67 76 84 90 92 53 72 58 76 52 64
1) 1)
1) 1) 1) 1) 1) 2) 1) 2)
1) 2) 1) 1) 1) 2) 1)
I11 (1) (2) 11) 11) (2)
63
70 71 81 74
822 (21 (3) (3) 131 (3) 141 131
(31 131 131
131 (2) (3) 12) (4) (2) (3)
(2) (31
12) (3) 12) (3) (3) 12) (4)
54 54 55 54 62 77 104 56 58 60 65 51 46
77 61
59 55 108 78 51 47 75
833
11) (2) (2) 12)
12) (2) (2) (2) 12)
(2) (21 (1) (2) (2) (2) (1) (2) (2) (3)
(1) (2) (2) 63 ( 2 ) 74 12) 56 ( 2 ) 121 ( 3 )
15 ( 1 ) 15 ( 1 ) 16 ( 1 ) 17 ( 1 ) 16 ( 1 ) 16 1 1 ) 28 1 1 ) 19 ( 1 ) 15 ( 1 ) 16 ( 1 ) 20 ( 1 ) 21 1 1 ) 19 1 1 ) 27 ( 1 ) 38 ( 1 ) 15 I l l 15 11) 23 1 1 ) 21 1 1 ) 21 1 1 ) 20 ( 1 1 29 ( 1 ) 28 (11 19 ( 1 ) 31 1 1 ) 29 1 1 )
812
813
823
14 ( 3 ) -3 ( 4 ) -1b ( 4 ) -6 ( 4 ) - 1 1 14) -46 161 -59 15) -12 ( 4 ) -3 ( 5 ) -1 (4) 10 ( 5 ) 11 ( 3 ) 26 141 33 ( 4 ) 0 (51 4 13) 10 141
12 ( 2 ) 4 (3) 2 (3) -1 1 3 ) -5 ( 3 ) -2 (4) -47 ( 3 ) -16 131 2 13)
3 -7 -5 1 -3 -4 -15 -10 3 -2
-b
(4)
25 -8 12 13 -12
15) (3) 14) 14)
4 6
(5) 131
9 -4 14 9
13)
(4) 12)
14) 31 131
11 (4) 4 12) -8 131
-18 -2 -1 -6 6 -7 14
(3)
12) (4)
12) 13) (3) 14) 14)
-2 1 3 ) -22 ( 4 )
10 I61
-3 -6
3
-18 -20 -7 -2 -29 -13 -14 -7
(1) (2) 12) (21
(2) (2) (2) (21 12) (2) (2) (2) 12) (2) (3)
(2) (2) 12) 13) (2)
(2)
-33 ( 2 )
-14 20 12 26
13) (21 (2) 13)
The form of the anisotropic temperature factors is exp[-Bllh* - B&2 - Ri,i* - Blah/? - B13hl - B*3kl]. and configuration at the five carbon centers. The enantiomorph assignment was made on the basis of the known absolute configuration of glucosamine, which is the acid degradation product of 4. Final atomic coordinates and thermal parameters far nonhydrogen atoms are given in Table I, and bond lengths and angles are listed in Tables I1 and 111. Table IV gives hydrogen coordinates.13 The furan ring is in a twist conformation rather than an envelope. In the view shown in Figure 1 , O ( l )is up and C(2) is down from the plane on the other three ring atoms. The oxazolidine ring and the acetyl on N(5) are nearly planar; torsion angles range from 3 to 10". The 0.1 M sulfamic acid degradation product ( 5 ) of 1 was reported in an earlier publication,l and the structure 14 was originally proposed. This structure has the a configuration, but there was no specific discussion of chirality at C-1. In a subsequent publication12 an isomer of 5 was reported whose spectral data and rotation suggested for it the structure 14. In view of this it was hypothesized that 5 was the CY isomer 15. H CHOH
/
4
v
Figure 1. Numbering, conformation, and configuration of 4. The configuration is 2S,3R,4R,8R,9R.
CH OH
HO
SCH
Y
12
OCOCH
0
15
CHIO
