Synthesis of the 3,3'-dinitro and 3,3'-diamino derivatives of a,a-trehalose and its D-gluco,~-marzno and

D-manno ,D-maitrzo isomers'

HANS H. BAER A N D A N N A J . B E L L Dc~prrr.rt?lent qf 'Chet?~isrt~, Ut7iver.sitj o,f'Otta,r.cr, 0ttuir.a. O t ~ t . , C~lniidn KIA' YB4

Received July 12, 1978

HANS H. BAER and ANNA J. BELL. Can. J. Chem. 56.2872 (1978) The tetraaldehyde 2 obtained frorn ?,?-trehalose (1) by periodate oxidation was cyclized with

nitromethane in the presence of sodium methoxide to give a mixture of 3,3'-dideoxy-3,3'-dinitro disaccharides in good yield. Acetalation of the product with benzaldehyde followed by chroma- tographic separation of the bisacetals afforded crystalline 4,6;4',6'-di-0-benzylidene derivatives (3, 4, and 5) of 3-deoxy-3-nitro-r-D-glucopyranosyl 3-deoxy-3-nitro-?-D-glucopyranoside (6, 3,3'-dideoxy-3,3'-dinitro-?,?-trehalose), 3-deoxy-3-nitro-a-~-glucopyranosyl 3-deoxy-3- nitro-z-D-mannopyranoside (7), and 3-deoxy-3-nitro-r-D-mannopyranosyl 3-deoxy-3-nitro-r- D-mannopyranoside (8), respectively. Acid debenzylidenation then gave crystalline 6-8, and these were hydrogenated catalytically to produce the corresponding dialnino disaccharide dihydrochlorides 9-11.

HAXS H. BAER et A X ~ A J. BELL. Can. J. Chern. 56,2872 (1978). Le tetraaldehyde 2, prepare par oxydation au periodate de l'z,z-trehalose (11, a ete cyclise

avec le nitromethane en presence de methanolate de sodium. On a obtenu un ~nelange de 3,3'-didesoxy-3,3'-dinitro disaccharides avec un boll rendenient. Par acCtalation du melange avec le benzaldehyde suivie d'une separation chromatographique des his-acetals on a isole a l'etat cristallin les derives 4,6;4',6'-di-0-benzylidene (3, 4 et 5 ) du (3-desoxy-3-nitro-5-D-gluco- pyranosyl)-3-desoxy-3-nitro-a-~-glucopyra1oside (6), du (3-desoxy-3-nitro-z-D-glucopyrano- syl)-3-desoxy-3-nitro-z-D-mannopyranode (7) et du (3-desoxy-3-nitro-r-~-mannopyra11osyl)- 3-dksoxy-3-nitro-cr-u-mannopyranoside (81, respectivement. L'hydrolyse acide des groupelnents benzylidenes a conduit aux composes cristallias 6-8, qui, par hydrogenation catalytique, ont donnC les dichlorhydrates des diamino disaccharides correspondants (9-11).

The disaccharide a,%-trehalose (c~-~-glucopyrano- syl a-D-glucopyranoside, 1) is widely distributed in nature and has been isolated from such diversified sources as bacteria, fungi and yeasts, algae, insects and other invertebrates, and also fro111 some higher plants. Its chemistry (2) and Inore recently its metabolisni (3) have been reviewed. The biological significance of the disaccharide is emphasized by its varied roles as a structural component (e.g., in the cord factor of inycobacteria), as a reserve supply of energy (e.g., in insects during flight), and as a possible iliterliiediate ill glucose resorption. Among the facets that continue to draw much attention is the mech- anism of action of the hydrolytic enzyme, trehalase, which is present not ollly in the types of organisms mentioned but also in mammals including man. Further interest in trehalose cheinistry arose from the discovery of the 2-amino-2-deoxy derivative (tre- halosamine) (4) and 2-ainino-2-deoxy-a-~-gluco- pyranosyl a-D-mannopyranoside (5) as antibiotically active metabolites of Streptotnyces species.

In continuation of our synthetic work (1, 6) on nitrogenous carbohydrates of potential biological

'Part 18 in a series on cyclizations of dialdehydes with nitromethane. For Part 17 see ref. 1.

interest we have decided to attempt the preparation of some new trehalose-type sugars for possible evaluatio~i ill biochemical studies. This was prompted by a suggestion of Dr. J . Defaye (Grenoble) whose laboratory in collaboratio~l with that of J. E. Courtois (Paris) is engaged in detailed studies (7-1 1) of structure-activity relationships in the mechanisin of trehalase action, for which there is a need for structurally and stereochemically modified trehalose analogs to be evaluated as substrates or inhibitors. A number of such co~npounds have been synthesized and assayed jointly by the French teams (7-1 1) while cornpreheiisive work on the chemical modificatioi~ of the disaccharide has also been performed by a group of British authors (120-j). Several synthetic amino, diamino, and tetraamino analogs have been described. These include the 2-amino-2-deoxy (13) and 6-amino-6-deoxy (14) derivatives oftrehalose and the 3-an1ino-3-deoxy-~-crltro isomer (12e) as well as 6,6'-diamino-6,6'-dideoxytrehalose (15), the 2,2'- diami110-2,2'-dideoxy-~-ultr.o:~-0ltro isomer (12e), the 4,4',6,6'-tetraamino analog of D-~O~OC~O,D-~O/OC~O configuration (129, and various 2,2', 3,3' , 4,4', 6,6', and 4,4',6,6' dialnines and tetraamines having additional deoxy functioils (12d, ,f-11). However, n o nitro derivatives of %,x-trehalose or its stereoisomers

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BAER AND BELL

H0 I I C H 2 0 H O - Q

H 0 OCH

OH

1 1 2

mixture of nitro disaccharides

I

OCH OCH:

P I1 Ph

Ph NO2 OH

h i q 0 S - ph<;dO OH

3 4 5

are known although we previously prepared (16) a structurally related nitro disaccharide (and the corresponding ainine) which contained a f3-linked heptuloside moiety. We have now employed the saine method, starting with 1, and obtained 3,3'- dideoxy-3,3'-dinitro-@,a-trehalose and two stereo- isomers as well as the correspoilding diamino sugars.

The tetraaldehyde 2, quantitatively produced (17)

fro111 commercial 1 by periodate oxidation, was doubly cyclized with nitroinethane in the presence of sodium methoxide according to the general procedure previously elaborated for sugar dialdehydes (18-20). Although a large number of stereoisomeric nitro sugars can theoretically arise in the reconstitution of t ~ v o pyranose ri11.g~ by this reactioii, it was expected 011 the basis of earlier experieilces that the D-gluco

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2874 CAN. J . CHEM. VOL. 56. 1978

TABLE I . The 100-MHz nnir data of compounds 3-8. Chemical shifts (6, fro111 TMS), w ~ t h coupl~ng constants (Hz) in parentheses

Compound Sol\ent H-l H-2 a H-3 H-4 H-5, -6, -6 PhCH Ar OH-2h Others

OMultiplicities refer to spectra after D,O exchange of OH-2 bSignal disappeared after D,O exchange. Cupper row, plucv ~no ie ry ; Ioaer row, r7ianr1o moiety. dSig~ial included in ui?resol\ed multiplets of 6 4 region. eSix-proton intensity; DOH due to deuterium euchange. fTetrahydrofuran. S igna l s disappeared on D 2 0 exchange (OH).

and D-n~mrrro configurations n ould be favored under the collditiorls of kinetic co~itrol that were eniployed (16, 18, 19). This proved in fact to be the case. The nilxture of nitro d~sacchar~des forliied in hlph yleld consisted ch~efly of three major coi~~ponents that \+ ele ldentlfied as 3-deoxq -3-n~tl o-cr-D-glucopyrano- syl3-deoxy-3-i11tlo-~-~-glucopyranos1de (6), 3-deoxy- 3-i11tro-a-~-glucop) ra11osyl3-deoxy-3-111tro-a-~-man- nopyranos~de (7), and 3-deoxy-3-n~tro-X-D-manno- p~lranosyl 3-deoxy-3-n1tro-~-~-mai1nopyranoside (8). They \\ere accornpa~lied by mlnor proportions of other products. wh~ch were not ~dent i f ied .~ Slnce a dlrect separation of the products could not be achleved the niixtule byas be11zylidenated to glve the corresponding bls-4,6-acetals 3, 4, and 5 nhlch wele Isolated 111 crystall~ne form by preparat~ve th~n-layer chromatography The ylelds mere 22, 25, and 9': based on the crude acetal mixture which had been obtalned In 81"; yield When the D - ~ ~ L ~ ~ o , D - ~ ~ ? u ~ I ~ o bisacetal 4 \ \as treated br~efly \b~ th ethanollc potas- slum hydrox~de at room tempeiature, lt partially ~ilcurred e p ~ m e r ~ z a t ~ o n to the thern~odynani~cally mole stable D-gluco,~-gluco iso111e1 3 whlch was isolated in 34>ield although no attempts were made to optimize the conditions for this transforma- tion. Similarly, it \+as demonstrated by meails of tlc that the ~-n?nntlo,~-l??u11no isomer 5 is in the same fashion convertible into 4 and thence 3. This facile epin~erization, known (21) from studies in analogous

2Apart from other configurational isomers, by-products might, in this reaction, conceivably be formed by cross cyclization of 2, with a nitrolnetha~ie molecule linking C-2 and C-2' to give a six-membered ring, although there was no evidence for this to have occurred to any large extent.

monosacchal lde derlvat~ves Q here ~t was foulid to be birtually quantltat~ve under appropilate cond~tlons, \$as to seive at t h ~ s time only to co~roborate the configurational asslgnme~its of the products, lt could doubtless be used as an ald to llnprovlng the y~e ld of 3 In case the latter IS the prlnclpal target of the synthesis.

The configurations of 3-5 follobved clearly from ' H ninr spectra T\TO of the compounds, namely those having the hlghest and lowest mobllitles in tlc, gave relatively slmple spectra showing only one slgnal for each set of posltlonally ldellt~cal protons 111

the ti50 ~uonosaccharld~c moiet~es and thus pioved themselves to be symmetr~cally constituted dlsac- char~des (3 and 5) By contrast, the compoulld of ~ntermediate chromatograph~c nlobll~ty (4) exhlb~ted a more complicated spectrum 1x1 which most sets of poslt~onally ident~cal pi otons (except for the aromatlc ones) gave, as far as the signals could be discerned, an lndlvldual s~gnnl for each of the t ~ z o molecule halves, with dlffei lng che~iilcal shlfts and coupl~ng constants, thus illdlcat~ng an unsy in i~~en~ca l dlsacchar~de struc- ture (Table 1) Compou~ld 3 could readlly be asslgned the D-&'~~~CO,D-~/Z/LO configurat~oll on account of the signal character of H-3, the proton at the nltlo- substituted posltlo~l The slgnal mas a quartet con- talnllig t n o large spl~telilgs (9 0 dnd 10.5 Hz), re-

31n 3-deoxy-3-nitro sugar derivatives this signal is normally located without difficulty as we know from a large body of experience (22). Barring 0-acylation at other ring positions it occurs at Io~ver field than those of the remaining sugar protons, with the possible exception of an equatorial (Y- anomeric) H-l signal which, however, is distinguished by its multiplicity.

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BAER AND BELL 2875

quiring axial orientation for H-2, H-3, and H-4. In agreement therewith, H-2 gave a quartet (after deuterium exchange of OH-2) showing 10.5 and 3.6 Hz spiittings, with the latter corresponding to that present in the anomeric doublet. The analogous signals in 5, namely an H-3 quartet ( J = 10.5 and 3.5 Hz), an H-2 quartet (J = 3.5 and 1.2 Hz), and the anorneric doublet ( J = 1.2 Hz) indicated the D-

matitzo,~-mur?no configuration. In COITIPOUII~ 4, the lowest-field ring proton signal, a quartet with splittings of 10.5 and 3.5 Hz at 6 5.55, was attri- butable to H-3 of a D-nzant~o moiety, and although the signal of the adjacent tI-2 could not be used for confirmation as it \vas not resolved from other resonances upfield, it became discernible after debenzylidenation of 4 (i.e., in 7) and was then found to agree with the assignment. (Moreover, the D-gakacto configuration, the only alternative that would also have been cornpatiblc with a similar H-3 quartet, \ \as ruled out by the aforementioned alkali- induced epimerization of 4 to 3.) Another H-3 signal was located a t 6 5.08 as a quartet having two large splittings (10.5 and 9.5 Hz) indicative of the D-gluco configuration of the second nlonosaccharide moiety. Again, this was confirmed by the spectrum of 7. As a point of incidental interest the chemical shifts of the benzylidene niethine protons may be compared (Table 1). I t has been-observed (23) in 4,6-0-benzyli- denated methyl 3-nitrohexopyranosides that this shift is a fairly sensitive indicator of over-all con- figuration. Thus, in DMSO-d6 solution, it occurred in the illethyl x-D-glucoside 0.1 ppm to higher field than in the methyl sc-D-inannoside. This phenomenon recurs in 3-5. Finally, it may be worth noting that the liydroxyl protons produced doublets with J = 9-10 Hz in gluco moieties (3 and 4) and J = 5 Hz in mutino moieties (4 and 5). In summary, a comparison of the three spectra reveals close similarities of the signals present in 4 with the respective ones in 3 and 5, as should be expected in view of the configurational relation. However, the spectrum of 4 is not an exact superposition of those ofits symmetrical isomers. The most notable divergence is associated with the shift position of the H-3 and H-3' protons in 4: in the gluco entity it is shifted upfield, and in the rnutlno entity, it is displaced downfield (by about 0.2 ppm each way), relative to H-3 in the corresponding sym- metrical isomers.

The bisacetals 3-5 upon de-0-benzylidenation by trifluoroacetic acid furnished the crystalline nitro disaccharides 6-8 in yields of 72-85%. The 'H nmr spectra of these pure sugars provided full confirma- ticn for the configurations. In every case the signals of H-1, H-2, and H-3 were amenable to analysis and accorded with the expected patterns (Table 1). The

mo1ecula~- rotations of 6-8 show a good corre- spoiidence ~ i t h those of the nonnitro analogs. In order to make this comparison, we took cognizance of the fact that substitution of a nitro group for 3-OH in methyl x-D-glucopyranoside increases the [MI, value by 6370 ~ h e r e a s such substitution in methyl Y-D-mannopyranoside decreases it by 5410. and we applied these increments" to the molecular rotations of r,x-trehalose and its N - D - ~ / L ~ c o , ~ - D - ~ ~ u I ~ / ~ ~ and Y - D - ~ l U t ? l l ~ , N - D - ~ ~ ? U n 7 7 0 isomers, for each glycosyl unit present (Table 2). The values so calculated for 6-8 deviate from those found by 3-8";.

An interesting feature of the nitrodisaccharides 6-8 - \\.as their propensity for retaining solkent of crystal- lization nhich i\as difficult to remove by heat~ng 111

~rrcuo. Thus. 6 and 7 wcrc obtalned as solvates incorporating tetrahydrofiiran, and 8 crystallized nith chloroform. The organic solvent could be dislodged by evaporation mith kkatcr, but highly stable hydrates were then obtained (see Experi- mental). Catalytic hydrogenation of 6-8 over platinum in the presence of a stoichiolr~etric amount of hydrochloric acid afforded the correspond~ng d~ailllno sugar dihydrochlondes. 3,3'-diamino-3,3'- d~deoxy-Y,Y-trehalose d~hydrochlonde (9) and its gluto,muti?zo (10) and mut~no,r~zan~?o (11) Isomers, as allalytically pure sol~ds.

NOTE ADDED ~h PROOF: Dr. J. Defaye kindly arranged to have co~npounds 6-11 111 20 rnilllmolar cor~centrat~on tested for hydrolys~s by cockchafer trehalase and reported that they s h o ~ no enzyme affinity. This is in accord \\ith recent find~ngs (7) ~ h i c h ind~cate that at least one unmod~fied v-D- glucopyranosyl moiety must be present for recog- nition.

Experimental Melting points were taken in glass capillaries in an elec-

trically heated alun~inunl block apparatus and are not cor- rected. Optical rotations were determined at room temperature in a Perkin-Elmer 141 automatic polarimeter. Infrared spectra were obtained from KBr discs in a Unicam SP 1100 spectro- meter. The nmr data refer to 100-MHz spectra with tetra- methylsilane lock signal, obtained on a Varian HA-100 instrument, and the coupling constants were measured at 250 Hz sweep width (Table 1). For qualitative tlc, precoated silica gel plates SIL G-25 UVZ5, (Macherey-Nagel & Co, West Germally) wcre used and the spots were made visible by spraying with 5% sulfuric acid in ethanol, followed by heating. Preparative tlc was performed on 18 cm x 40 cm glass plates coated with a 1.5-mrn layer of MN Kieselgel G/UV254 (Macherey-Nagel & Co.). Unless otherwise indicated, solvent systems en~ployed for developn~ent were (A), 3 : 1 chloroform- methanol; (B), 4: 1 chloroform - ethyl acetate; and (C), 7 : 3

4The data are tied to the configurations from which they were derived; comparison of other glycosides with their respective 3-nitro derivatives reveals effects 011 molecular rotation that are similar in trend but different in magnitude.

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CAN. J . CHEM. VOL. 56, 1978

TABLE 2. Molecular rotations

Mol. -

Compound weight [E], in FI,O (ref.) Observed Calculated

Methyl r-D-glucopyranoside 3-dcoxy-3-nitro derivative

Methyl r-D-mannopyranoside 3-deoxy-3-nitro derivative

a-D-Glucopgranosyl x-~-gl~1copyranoside~2H~O 3,3'-dideoxy-3,3'-dinitro derivative.H,O

x-D-Glucopyranospl a-D-mannopyranoside0.5 H,O 3,3'-dideoxy-3,3'-dinitro derivative.THF

r-D-Mannopyranospl r-D-mannopyranoside 3,3'-dideoxp-3,3'-dinitro derivative

ether - petroleurn ethel. Here and else\+here In the text. petroleum ether refers to the fraction boilmg at 30-60 C.

Periodate Oxidntiorr o f x,r-Ti~elicrlose ( l j To a solution of r,x-trehalose dihydrate (1) (2.05 g, 5.42

rnmol) in water (20 ml) was added a solutio~l of sodiurn meta- periodate (4.87 g, 22.7 rnmol, i.e., a 5% excess over 4 molar equiv.) in water (50 nil). The reaction mixture was kept in the dark at room temperature for 24 11, during which the pH value of the medium was periodically adjusted to approximately 6 by the portion\vise addition of sodium hydrogen carbonate (0.89 g) in water (20 nil). Examination by tlc (double irriga- tion with solvent A) showed that the very slorv-moving 1 was c~entually replaced by a single, fast-moving product, the tetraaldehydc 2. Several spots having intermediate mobilities were seen during early stages of the oxidation and were attributable to reaction intermediates. For processing, ethanol (30 ml) was added to the reactioli mixture and, after cooling with ice water: the inorganic precipitate was filtered off and ~vashed with chilled ethanol. The filtrate was concentrated to a small volurne by evaporation and fresh ethanol (30 ml) was added to precipitate further inorganic material, which was remoled. This operation was repeated three or four times, wcth evaporations being conductcd to near dryness, until eventually the oxidation product dissolved clearly and was then obtained as a dry, uhite foam upon solvent evaporation; yield, 1.70 g of 2.

Nifrorneflicne Cj~clization of 2 The tetraaldehyde 2 (1.70 g) and nitromethane (1.0 ml) were

dissolved in absolute methanol (20 ml), and the mixture was rendered alkaline to pH 10 (measured potentionietricallp) by dropwise addition of a dilute, methanolic sodium methoxide solution (freshly prepared from sodium hydride; c, 0 . 5 z Na). It was important to avoid an excessive amount of base as this tended to give impure, discolored products; 011 the other hand, too low an alkalinity caused iriconveniently slow reaction. The mixture, which became deep-yellow and deposited some precipitate (nitronate salts), was allowed to stand in a closed vessel at room temperature for 3.5 h. It was then deionized by stirring with a cation exchange resin (3 ml of Amberlite IR- 120-H' or equivalent) whereby the precipitate dissolved. The resin \\as filtered off and washed with methanol, and the filtrate was evaporated to give a dry, yellow solid of nitro sugars (2.0 g, about 9OW< based on 1). The product mixture was dissolved in ethyl acetate and recovered as an off-white powder (1.67 g, 77%) by precipitation with anhydrous ether.

Beniylidenntioiz otid Separation The above mixture of nitro sugars (1.67 g), benzaldehyde

(10 ml), and finely divided anhydrous zinc chloride (2.5 g) were

placed under a nitrogen atmosphere in a closed vessel pro- tected fro111 light aud \'igorously stirred at room temperature for 4.5 dabs. The reaction mixture was then poured into, and agitated with, crushed ice and an equal amount of petroleum ether. The solid organic product which separated \\as col- lected, washed \\ell with petroleunl ether, aud dried in the air. It mas then dissol\ed in ether and recokered by precipitation, under ice cooling, with chilled petroleurn ether. The pale yellon material (1.96 g, 81%) showed three well-separated, major spots in tlc. By triple irrigation with solvent B on a 9-cni plate, these migrated approxin~ately 6, 4, and 2 cm. There were also several minor spots between the fastest and the second, and just above arid below the third ruain spot. The main spots represented conipounds 3, 4, and 5, in order of decreasing mobility. These products were then isolated by preparative tlc using two to four successive irrigations with solvent B. Seven plates of 18 crn x 40 cni were used to accomniodate 1.82 g of the product mixture. There uas obtained, in almost pure condi- tion, crystalline 3 (0.40 g) and 4 10.46 g), which represented 22 and 257 , respectively, of the mixture applied. The third frac- tion, which consisted mainly of 5, was obtained as a gum (0.34 g, 18.7z) that was revealed by tlc (solvent C) to contain small proportions of several contaminants. It yielded crystal- line 5 upon purification by a second, preparatile tlc (solvent C). Material moving even more slowly than 5, isolated in the first chromatography and amounting to 0.6 g, was inho~no- geneous and not investigated further.

4,6-O-Be11zy/idci1e-3-deo~vy-3-nifro-x-~-g/rtco~~yrnr1o~cy/ 4,6-0- Be11zylidcne-3-deo~~j~-3-nifro-~~-~-glrcopynoside f 3 j

Recrystallized from chloroform, 3 consisted of fine, colorless needles, mp 259'C (dec.); [r], f 11 1.6: ( c 0.4, acetone); v , , , ~ ~ ~ ' ~ : 3580 (OH), 1550, 1360 (NO,), and 760, 703 cn1-' (benzylidene). Anal. calcd. for CZ6HzsPc'2013 (576.5): C 54.16, H 4.90, N 4.86; found: C 54.05, H 4.86, N 4.80.

4,6-O-Benz~lidene-3-deoxy-3-nitro-x-~-glrrco~~yra11o~yl 4,6-0- B e n z ~ . / i d e 1 1 c - 3 - d e o x ~ ~ - 3 - n i t r u - . r - ~ - ~ ( 4 )

Recrystallized from chloroform, the fi~ie colorless needles of 4 deconlposed at 225'C; [r], + 81.5" (c 0.5, acetone); v,,,,("~'): 3450 (broad, OH), 1552, 1365 (NO,), and 760, 703 cm-' (benzylidene). Anal. calcd. for C~6H28f\/2013 (576.5): C 54.16, H4.90, N 4.86; foulid: C 54.07, H4.81, N 4.96.

When a sample of 4 (900 mg) was dissolved at room tem- perature in 12 ml of 0.13 M ethanolic potassium hydroxide solution and, after 15 min, deionized by cation exchange, the isomer 3 was a prominent product among remnant 4 and some minor decomposition products, according to tlc (solvent B). By preparative tlc of the reaction mixture, 260 mg of pure 3 could be isolated. The remainder of the material recovered from this operation was subjected to a second, identical

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BAER AND B E L L 2877

treatment with base, and another 50 lug of 3 could subse- finished within 2 h. The catalyst was removed and washed well ~ u e n t l y be isolated. ~vith at least seven 10-ml portions of water; the uroduct

4 , 6 - O - B e n z y I i i l e 1 r e - 3 - c l e o x y - 3 - ~ 1 4,6-0- ~eizz~~/ide11e-r'-deo,~~~-3-r1ilt~o-x-D-mnnnop)~rn1~o~i~~e ( 5 )

Conlpound 5 as obtained froni the chromatography was recrystallized from ethanol - ether - petroleum ether to give fine needles (0.16 g), mp 175-176'C; [a], +51.4' (c 0.4, acetone): v,,,'~ Br): 3550 (broad, OH), 1552, 1365 (NO,), and 765, 705 cm-' (heezylidene). .4na/. calcd. for C,6H28N,01, (576.5): C 54.16, H 4.90, N 4.86; found: C 53.98, H 5.04, N 4.74.

Compound 5 was shown by tlc (solvent B) to epimerize to a nlixture of 3 and 4 by brief treatment with 0.1 M ethanolic potassium hydroxide followed by deionization.

shoued a considerable tendency to remain adsorbed. The combined aqueous solution and washiilgs were evaporated to dryness to give the amino sugar dihydrochloride 9 (140 mg) as a white solid. The isoiners 10 and 11 were obtained in the same way from 7 and 8, respectively. The products gave single spots, indicated by ~iinhydrin spray, in paper chromatography (descending, on Whatman No. I paper) using the Fischer- Diirfel (28) solvent system, 5 : 5 : 3: 1 pyridine - ethyl acetate - acetic acid - water, with 11 :40: 6 pyridine - ethyl acetate - water in the bottotli of the tank. The R,,.,,,, values were 0.43 (9), 0.39 (lo), and 0.35 (11). Occasionally, somewhat lower values were found owing to minor fluctuations in solvent com- position or temperature, but the ratio was remarkably con- stant for values obtained in the same run.

Each of the acetals 5-5 (0.35-1.0 g) was treated with 90% trifluoroacetic acid (27) at 0 : ~ for 0.5 h. =he acid (5 ml) was 3-Anriflo-3-n%ox~-a-~-ghtcopj~ranosy/ 3-~l?liilo-3-c/eoxy-a-D-

removed by evaporation in caccto followed by coevaporation glr~copyrcrnoside Dihydrochloride (9 ) mith several successive portions of ethanol. The residues were The crude product was in

as stated to furnish pure 6 , 7, and 8 ill minimuni amount of methanol, recovered by careful precipita- yields of 77, 72, and 85%, respectively. tion with ethyl acetate, dried in cacno at 56'C for 4 h and then

represented a somewhat hygroscopic, fine uhite 13ouder 3-Dtoxy-3-nilro-a-~-g/11~0[?):1~~111o~~y/ 3-Deosy-3-nitro-x-D-

glucopyrnrroside 1'6) Recrystallized from tetrahydrofuran-ether, the white

needles of 6, n1p 167-169'-C, contained 1 mol of tetrahydro- furan of crystallization as evidenced by the nrnr spectrum (multiplets of 4H intensity at 6 1.80 and 3.64). This sol\ent could not be removed in cucrro at 110'C unless the analytical sample was first treated with a small amount of water and then dried (4 h, 110-C). However, 6 then appeared to exist as a monohydrate, according to the elemental analysis. It showed mp 159-16IoC, [a] , t 187- (c 0.3, water), v , , , (~~ ' ) : 3700-3300 (very broad, OH), 1555, 1370 cnl-' (NO,). Annl. calcd. for C,,H,,N,O,,~H,O (418.3): C 34.45, H 5.30, N 6.70; found: C 34.67, 34.39, H 5.25, 5.28, N 6.61, 6.72.

3-Deoxy-3-nitro-a-u-gl~rcop~~ra11o.sy 3-Deoxy-3-~iitio-x-D- ~nat~nol,)'ralzosideyraroside / 7)

After repeated recrystallizations from tetrahydrofurall-ether, 7 was obtained as a solvate with tetrahydrofuran, according to the nmr spectrum and elemental analysis. It showed mp 145- 147"C, [%ID A 133' (c 0.3, water), v , , , , ~ ~ ~ ' ) : 3700-3300 (very broad, OH), 1560, 1365 cm-' (NO?). The analytical sample \%-as dried it1 cncnu at 1lO'C for 4 h. A~rnl. calcd. for C,,H,,- NZOI3.C4H80 (472.4): C 40.68, H 5.98, 5.93; found: C 40.81, H 6.01, N 5.85.

When another analytical sample was evaporated with added water before drying it lost the tetrahydrofuran b~:t analyzed as a hemihydrate. Annl. calcd. for C,,H,,N2OI3~0.5 H 2 0 (409.3): C 35.21, H 5.17, N 6.85; found: C 35.19, H 5.33, N 6.50.

3-Deo.~y-3-1rilro-x-~-mnntrop)~rnno~y 3-Deoxy-3-nitro-x-D- n~arznopyrat~oside ( 8 )

Compound 8 was recrystallized froni chloroform-methanol; nip 185-192°C (dec.), [%I, f 83' (c 0.3, water); ~ , , , (~~ ' ) : 3700- 3300 (very broad, OH), 1550, 1370 cin-' (NO,). The analytical sample, dried in cacno at 110'C for 4 h, retained chloroform of crystallization, which was confirmed by its nmr spectrum in DMSO-d6. Anal. calcd, for C,,H,,N,O,,~~CWCI, (440.05): C 33.65, H 4.66, N 6.37; found: C 33.66, H 4.98, N 6.37.

(pbssibly a micro crystal line^ hydrate) that decomposed at 230cC; [r], + 168' (c 0.3, water); v,,,'"~'): 3500-3100 (OH, NH,-), -1620 and 1510 cm-' (NH,-). For analysis, a sanlple was dried further, at 80'C and 1 Torr, whereby it lost 6 .525 of its weight. Anrtl. calcd. for C,,H,,CI,N,O, (413.3): C 34.87, H 6.34, N 6.78; found: C 34.97, H 6.46, h- 6.67.

3-Ai?1itro-3-deo.~)'-x-~-g/ucop)'rnno'I 3-Arnirio-3-deosy-x-n- ~nnnnopyrunosiclr Dihydrocllloi.ide (10)

Prepared in the same way as jusi described for 9, the isomer 10 was similar in appearance and in its ir data; it decomposed at 20ScC and showed [x], + 124.5" (c 0.3, water). Dried to constant weight at 80-C and 1 Torr, the analytical sample lost only 3.947, of its weight and then gave values corresponding to a monohydrale. Antrl. calcd. for C12H,6CI,V,0,.H,0 (431.3): C 33.42, H 6.54, N 6.50; found: C 33.21, H 6.45, N 6.41.

3-Anz0~o-3-deoxy-x-~-n7nrrnopyrir1ro.1y/ 3-An~ino-3-fleo.uy-r-~- mcirrnopyranoside Dillydrocirloride (11)

The crude product 11 was purified by ctissolution in methanol and careful precipiiatio~l with ether. The dry white powder showed similar ir bands as 9. It decoinposed at 200-202'C and had [.J], + 84.3- (c 0.3, hater). When dried to constant weight at 100-C and 2 Torr, it still retained water and analyzed as a dihydrate. Anrrl. calcd. for C,,H,6CI,N,0,~2H,0 (449.3): C 32.08,H6.73,N 6.24;found: C32 .00 ,H6 .55 ,N6 .15 .

Acknowledgments

This work was supported financially by the National Research Council of Canada. H.H.B. a-ishes to express his appreciation to Dr. Jacques Defaye for generous hospitality and stimulating discussions during a sojourn at his laboratory in the Centre de Recherches sur les MacromolCcules VCdtales, C.N.R.S., Grenoble. France. where the

CntaIytic Hydrogenations wbrk was conceived. ~ l ; e ~a t ion 'a l Research Platinum dioxide catalyst (200 rng) suspended in 0.5 M council of canada alld the celltre ~ ~ ~ i ~ ~ ~ ~ l de la

hydrochloric acid (2 ml) was prehydrogenated and a solution of 6 (160 lllg) in 0. hydrochloric acid (4 rill) was then Recherche Scieiltifique are thanked for the award of a introduced. With efficient shaking under hydrogen at room travel stipend under the auspices of their scientific temperature and ordinary pressure the hydrogenation was exchange program.

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2878 CAN. J . CHEM.

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