the structure and the properties of acetone ... · pdf filethe structure and the properties of...

THE STRUCTURE AND THE PROPERTIES OF ACETONE- METHYLRHAMNOPYRANOSIDE

BY P. A. LEVENE AND IRVING E. MUSKAT*

(From the Laboratories of The Rockefeller Institute for Medical Research, New York)

(Received for publication, March 27, 1934)

The work now communicated constitutes a part of the general plan for the preparation of partially substituted pentoses. In nature they occur as component parts of nucleic acids in the form of esters of phosphoric acid. It was especially desired to develop methods for the preparation of monophosphoric esters of pentoses having a definite known structure.

In the case of glucose the key substance for the preparation of partially substituted derivatives has been monoacetone-glucose. In the case of rhamnose there are, a priori, two different types of monoacetone derivative possible-one having a furanose and the other a pyranose structure. The structure of monoacetone rham- nofuranose has been established by Freudenbergl as the 2,3- monoacetone derivative, thus leaving position (5) open for partial substitution. The position of the free hydroxyl group in the monoacetone-methylrhamnopyranoside might be in any position other than (1) and (5). It has now been found that in the acetone derivative of the methylrhamnopyranoside the acetone residue also occupies positions (2) and (3) (II), thus permitting the preparation of derivatives substituted in position (4). Of these, 4-methylrhamnopyranose (III) and 4-p-toluenesulfonyl-rhamnopyranose (IV) have, so far, been prepared.

The former of these two substances served for the elucidation of the structure of monoacetone-methylrhamnopyranoside. On oxidation with silver oxide according to the procedure of Freuden- berg,’ the silver salt of methyldihydroxybutyric acid (VII) was obtained. The formation of this substance was sufficient. evidence

* National Research Fellow in Chemistry. I Freudenberg, K., Ber. them. Ges., 69,836 (1926).

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432 Acetone-Methylrhamnopyranoside

----CH-OCH,

I H-C-OH

H-b-OH

HO--C-H I

I C&

I

-CH- OCR, YCI-T-OCH,

I I H-C-O CHB R-C-O

‘C/ \C/CH3

/\ 0 /\ H-C-O CHa CHs

I HO-C-H

I ---C-H

I CHs C&

II III

r CH-OCH,

I H-C-O C&

‘Cd 0 /\ 6

H-C-O C&

I C~HTO~SO-C-H

I L C-H

I CK

IV

__ CH-OH CH-OCOCHs

I H-C-OH H-C-0-COCH,

H-C-OH H-C-0-COCH:j

I I H&O-C-H HICO-C-H

I I -C--H L-C-H

I I CHs C& V VI

-----CH--Br

H-C-OCH,

I H-C-OH 0 H-C-0-COCHa 0 H-C-0-COCHa

I I CH, HaCO-C-H

I L-------C-H

I

CH:, CHz

VII VIII IX


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P. A. Levene and I. E. Muskat 433

N-C-N-CHI YCH-0 OCH3

// HC

II I

‘C/

\ /\ i ----CH -N-C C=O H-C-O CHs

I II 0 I H-C-0-COCH, O=C-N-CH3 H-C-0-COCHz

I I 0 H-C-0-COCHz HaCO-C-H

I I HSCO-C-H ‘----C-H

I L-&H

I C& I

X XI

for the assumption that position (4) of the monoacetone-methylrhamnopyranoside was free in the origina. substance and had been substituted on methylation.

The p-toluenesulfonate was prepared for the purpose of possessing a derivative in which the group could be substituted by another, if so desired. Inasmuch as the final aim in the preparation of partially substituted pentoses was the synthesis of nucleosides, the bromoacetyl derivative of 4-methylrhamnose was condensed with silver theophylline (X). Incidentally it may be mentioned that evidence was obtained indicating the formation of an orthoacetate (XI) when this bromoacetate was treated with methyl alcohol in the presence of quinoline.

EXPERIMENTAL

Preparation of Methylrhamnopyranoside (I)-Methylrhamno- pyranoside was first prepared by Fischer2 who proved that, it exists in two crystalline forms: (1) m.p. lOgo, [a], = -67.2” (in water); (2) m.p. 140”, [o(], = +95.2” (in water). For this inves- tigation the methylrhamnoside was prepared as follows: 5 gm. of dry rhamnose were dissolved in 50 cc. of methyl alcohol containing 1.5 per cent of dry hydrogen chloride, and the solution was refluxed for 105 minutes. The solution was cooled, rendered neutral with silver carbonate, and the mixt,ure was filtered. The filtrate was

2 l+xhx, IL, Mw. clwru. Gs., 28, 1158 (1895).


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Acetone-Methylrhamnopyranoside

shaken with a little activated charcoal to remove traces of silver salts, and the mixture was again filtered. The colorless filtrate was evaporated to a thick syrup under diminished pressure, and the syrup was distilled. The entire product distilled without decomposition at 145-150” under 0.14 mm. pressure. No effort was made to crystallize the syrup and separate the OL and /3 forms. The substance had the following composition.

4.855 mg. substance: 8.462 mg. CO?, 3.380 mg. Hz0 5.358 “ “ : 7.190 “ AgI

C1HL405. Calculated. C 47.16, H 7.9, OCHI 17.42 178.1 Found. “ 47.50, “ 7.8, “ 17.71

Preparation of Monoacetone-Methylrhamnopyranoside (II)- 10 gm. of methylrhamnopyranoside were suspended in 200 cc. of acetone containing 0.2 per cent of sulfuric acid. Anhydrous cupric sulfate (20 gm.) was added and the mixture was shaken at 37” for 20 hours. The mixture was filtered and the filtrate rendered neutral by shaking with calcium oxide. The calcium sulfate and excess calcium oxide were removed by means of filtration and washed with small quantities of acetone. The filtrate was evaporated under diminished pressure and the residual syrup was distilled. The entire product distilled without decomposition at 110-112” under 1 mm. pressure. nz6 = 1.4533.

1t.s specific rotation was

[a]:: = -0.42” X 100

2 x 1.4850 = -14.1” (in water)

The substance had the following composition.

4.752 mg. substance: 9.589 mg. COz, 3.550 mg. Hz0 5.920 “ “ : 6.346 “ AgI

C,OH,SOS. Calculated. C 55.02, H 8.3, OCH, 14.22 218.1 Found. “ 55.02, “ 8.4, “ 14.14

Preparation of Methylmonoacetone-Methylrhamnopyranoside (III)-The acetone-methylrhamnopyranoside (21.8 gm.) was methylated by means of Purdie’s reagents (CHJ, 100 gm., and AgzO, 116 gm.) and the product isolated as a colorless syrup. The mcthylation was twice repeated with half the above quantities of methylating agents and the product was isolated as a colorless,


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mobile syrup which distilled at 67-68” under 0.3 mm. pressure. 26

nD = 1.4369. Yield, 18 gm. (77.6 per cent). Its specific rotation was

[al: = -0.25" X 100

2 X 0.5200 = -24.0" (in water)


4.160 mg. substance: 8.695 mg. COz, 3.060 mg. Hz0 7.915 “ “ : 15.990 “ AgI

C1lH,oOs. Calculated. C 56.86, H 8.7, OCH, 26.72 232.2 Found. “ 56.99, “ 8.2, “ 26.66

The methylation of monoacetone-methylrhitmnopyranoside was also carried out in liquid ammonia by the method recently described by Muskat. 3 Since the technique now used is different from that described in the preliminary paper, this methylation will be described in greater detail.

Monoacetone-methylrhamnopyranoside (21.8 gm., 0.1 mole) was dissolved in about 150 cc. of anhydrous liquid ammonia and 4.0 gm. (slightly more than 0.1 mole) of potassium, contained in a cylindrical cup made of platinum gauze, were suspended above the liquid ammonia solution of the sugar. Anhydrous liquid ammonia was then allowed to drop onto the potassium. The potassium readily dissolved in the liquid ammonia, and the solution so formed dropped into the sugar solution. Reaction took place imme- diately, as could be seen from the disappearance of the blue color of the solution of potassium in liquid ammonia. The mono- potassium salt of acetone-methylrhamnopyranoside was thus formed.4 After all of the potassium in the platinum cup had dissolved and reacted with the sugar, the flow of liquid ammonia was stopped. The potassium salt of acetone-methylrhamnopyranoside remained in solution (about 400 cc. of liquid ammonia).

The methyl iodide may now be added to the liquid ammonia solution of the potassium salt, as was described in the paper referred to above;3 however, it is more satisfactory to isolate the salt and carry out the reaction with methyl iodide in the absence

3 Muskat, I. E., J. AWL. Chem. Sot., 66,693 (1934). ,I The apparatus and procedure used in this work will Ix dcscrihed iu a

l~~tpcr by one of us (Musknt) to nppcar soon.


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of ammonia. The reasons for this changed procedure will be discussed in a paper by one of us (Muskat) to appear soon.

The ammonia was completely removed by vaporization under strictly anhydrous conditions. The potassium salt of acetone- methylrhamnopyranoside remained as a white crystalline material which was hygroscopic. The reaction vessel containing the salt was cooled in an ice-salt bath and 50 cc. of anhydrous methyl iodide were added slowly with constant shaking. Reaction occurred very rapidly and heat was evolved. After all the methyl iodide had been added, the reaction product was thoroughly shaken for 10 minutes. In order to insure that the reaction was complete and that all the potassium salt had reacted with the methyl iodide, a sample of the product was removed and dissolved in water. The aqueous solution was neutral, showing that the potassium salt had reacted completely.

The excess methyl iodide was now distilled from the reaction products and the residue was extracted several times with hot chloroform. The combined chloroform extracts were filtered and the chloroform was removed by vaporization. The residual oil distilled completely at 67-68” under 0.3 mm. pressure. Its physical properties and analysis showed it to be identical wit,h the methylacetone-methylrhamnopyranoside prepared by the Purdie method described above. Yield, 21.8 gm. (94 per cent). Besides giving a greater yield of product, the reaction is much more quickly carried out and is less expensive.

Preparation of p-Toluenesulfonylacetone-Methylrhamnopymno- side (IV)-p-Toluenesulfonyl chloride (1.6 gm.) was added to a solution of acetone-methylrhamnopyranoside (1 gm.) in dry pyridine (1.5 cc.). The mixture was allowed to stand overnight a,t room temperature and then dissolved in chloroform with t,he addition of water. The aqueous portion was extracted twice with small quantit>ies of chloroform. The combined chloroform extracts were then washed successively with dilute hydrochloric acid, dilute sodium hydroxide, and water. The chloroform solution was now dried over anhydrous sodium sulfate, filtered, and the solvent removed by vaporization under diminished pressure. The product was a pale yellow syrup which crystallized from ligroin (3040”) in white needles, m.p. 58”. The compound was very soluble in the usual organic solvents with the except,ion of ligroin.


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Its specific rotation was

[al:: = +0.294” x 100

2 x 1.0500 = +14” (in methyl alcohol)


4.906 mg. substance: 9.890 mg. CO+ 2.800 mg. Hz0 9.360 “ “ : 5.835 “ BaSOl

ClrH2407S. Calculated. C 54.80, H 6.5, S 8.60 372.26 Found. “ 54.97, “ 6.4, “ 8.56

2 gm. of the p-toluenesulfonylacetone-methylrhamnopyranoside were heated in a sealed tube for 2 hours with 2 gm. of sodium iodide and 10 cc. of acetone at 100”. The solution was then evaporated to dryness under diminished pressure, and the dry residue extracted repeatedly with hot chloroform. The chloroform extracts were washed with dilute sodium thiosulfate solution and then with water. The chloroform extract was dried and the solvent removed by vaporization. The resulting oil (2 gm.) was crystallized from ligroin (3040’) and melted at 58”. A mixture with the original p-toluenesulfonylacetone-methylrhamnopyranoside also melted at 58”. A qualitative test for halogen was negative. This is in accord with the previous finding that a p-toluenesulfonyl group attached to a secondary alcohol cannot be removed under these conditions.

Preparation of Monomethylrhamnopyranose (V)-The methylmonoacetone-methylrhamnopyranoside (14 gm.) was hydrolyzed with 2 per cent sulfuric acid (300 cc.) on the water bath during 20 hours. All the methylmonoacetone-methylrhamnopyranoside had dissolved within this time. The solution was rendered neutral with barium carbonate, filtered, and the water removed under diminished pressure, at a temperature not exceeding 40”. The residual syrup was dissolved in acetone and filtered from barium salts. The acetone was removed by vaporization under diminished pressure but the resulting yellow viscous syrup could not be distilled. In order to purify this syrup it was acetylated as described below.

Preparation of Triacetyl-Monomethylrhamnopyranose (VI)-10 gm. of monomethylrhamnopyranose were dissolved in a solution of 20 cc. of acetic anhydride in 25 cc. of pyridine. The solution was


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allowed to stand at 0” overnight and then poured onto ice with shaking. The ice-cold aqueous solution was extracted several times wit-h small quantit)ies of chloroform and t*he combined chloroform extracts were washed with dilute sulfuric acid, sodium hydroxide, and finally with water. The chloroform solut’ion was then dried over anhydrous sodium sulfate and the solvent removed under diminished pressure. The residual syrup distilled almost completely at 114-116’ under 0.17 mm. pressure. nz = 1.4452.


[cxl:: = -0.38” x 100

2 x 1.5600 = -12.2” (in methyl alcohol)


4.120 mg. substance: 7.760 mg. CO*, 2.345 mg. Hz0 4.885 “ “ : 3.800 “ AgI

69.50 “ “ required 6.9 cc. 0.1 N NaOH CdLo0~. Calculated. C 51.26, H 6.6, OCHa 10.20 304.2 Found. “ 51.36, ” 6.4, “ 10.26

Calculated, COCHJ 42.43; found, 42.69

Deacetylation of Triacetyl-Monomethylrhamnopyrano.se-lle- acetylation of the triacetyl-monomethylrhamnopyranose (10 gm.) was carried out by dissolving the substance in methyl alcohol (250 cc.) and adding one-thirtieth of the theoretical amount of barium methylate, and then allowing to stand at 0’ overnight. The solution was now saturated with carbon dioxide, 50 cc. of water added, and the solution again saturated with carbon dioxide. After the addition of a little characoal, the solution was boiled for a few minutes, filtered, and the solvent removed by vaporization under diminished pressure. The resulting glassy, syrup was crystallized from a small amount of methyl alcohol. It was recrystallized from ethyl acetate and finally from acetone containing a small amount of dry ether. The pure crystals melted at 122”. Its specific rotat’ion was

[a]; = ‘~‘~g~800 = f12.9” (in methyl alcohol)

roll; = -0.15” x 100

2 x 0.580 = - 12.9” (in methyl alcohol with addition of ammonia)

s Levene, P. A., and Tipson, R. S., J. Riol. Chem., 93,631 (1931).


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5.020 mg. suhstnnre: 8.700 mg. CO,, 3.550 mg. H,O 5.915 “ “ : 7.705 “ AgI

C;H,,OA. Calrulntcd. C 47.6, H 7.9, OCHa 17.42 178.1 Found. *’ 47.26, “ 7.9, “ 17.19

The phenylhydrazone was prepared in the usual manner and was isolated as yellow needles. It decomposed at 176”.

Oxidation of Monom.ethylrhamnopyranose-The monomethylrhamnopyranose was oxidized with silver oxide according to the method of Freudenberg.’ 3 gm. of the sugar were heated on the water bath overnight with 36 gm. of silver oxide suspended in 150 cc. of water. The reaction mixture was shaken frequently. The silver salts were then removed by means of filtration and the filtrate was concentrated to a small volume under diminished pressure. The solution was filtered while hot, and on cooling, crystals separated out. Absolute alcohol was added to hasten the crystallization. The pure white crystals were collected on a filter and dried.


[aI’d = -0.50” x 100

2 x 0.844 = -29.6” (in water)

The substance had the following composibion.

6.931 mg. substance: 6.335 mg. CO?, 2.250 mg. Hz0 5.970 “ “ : 5.705 “ AgI 6.931 “ “ : 3.110 “ Ag

GHsO,Ag. Calculated. C 24.90, H 3.8, OCHI 12.87, Ag 44.7 240.95 Found. ” 24.92, “ 3.6, “ 12.61, “ 44.9

Preparation of Bromodiacetyl-Monomethylrhamnopyranose (V111)-5 gm. of triacetyl-monomethylrhamnopyranose were mixed with 25 cc. of glacial acetic acid containing 40 per cent of dry hydrogen bromide.6 The resulting solution was allowed to stand at room temperature for 60 minutes, after which the hydrogen bromide gas was removed under diminished pressure at room temperature. The solution was then diluted with 100 cc. of toluene and evaporated to a thick syrup under diminished pressure

B Levene, I’. A., and Tipson, R. S., J. Riol. Chem., 92,109 (1931).


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at 35”. Two further portions of 50 cc. of toluene were run in and evaporated off. This syrup was now dissolved in 50 cc. of benzene and the solution was evaporated to a thick syrup. Traces of solvent were removed under a high vacuum at 35”.

The resulting thick, pale yellow syrup was dissolved in the minimum of cold dry ether, a further 5 cc. of ether was added, and then ligroin was added to incipient turbidity. A little charcoal was added and the mixture shaken and filtered on a fluted filter, the filtrate obtained being absolutely colorless. On cooling in ice, crystallization set in. The colorless crystals melted at 104.5’.


[a]:: = -7.16” X 100

2 x 2.0050 = -178.6’ (in chloroform)


9.995 mg. substance: 5.740 mg. AgBr (direct precipitation) CIIHITOGBr (325.05). Calculated, Br 24.6; found, 24.44

Preparation of Methyl-Diacetylmethylrhamnopyranoside (IX)- The bromodiacetyl-monomethylrhamnopyranose prepared from 4 gm. of triacetyl-monomethylrhamnopyranose was dissolved in 50 cc. of methyl alcohol. 10 gm. of dry freshly prepared silver carbonate were added and the mixture was shaken vigorously for 15 minutes, after which no bromine was found in a small filtered test portion. The mixture was filtered, the silver salts well washed with dry ether, and the combined filtrate and washings evaporated to a thick syrup under diminished pressure. It was dissolved in dry ether, treated with charcoal, filtered, and the solvent removed by vaporization. The syrup disClIed at about 125-130’ under 0.7 mm. pressure. nt4 = 1.4499.


[al:: = +0.95” x 100

2 X 2.76 = +17.2” (in methyl alcohol)


4.091 mg. mg. mg. substance: 7.785 COZ, 2.505 Hz0 6.170 “ “ : 9.482 “ AgI

293.0 “ “ required 20.65 cc. 0.1 N NaOH CdLo0~. Calculated. C 52.1, H 7.3, OCH3 22.46 276.2 Found. “ 51.9, “ 6.9, “ 20.2

Calculated, COCH, 31.15; found, 30.3


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Preparation of Methyldiacetyltheophylline-Rhamnopyranoside (X) -2 gm. of bromodiacetyl-monomethylrhamnopyranose were dissolved in 100 cc. of dry xylene. 2 gm. of the silver salt of theophylline, dried to constant weight at 140” under reduced pressure, were added and the mixture was allowed to stand on the water bath overnight, after which no bromine was found in a small filtered test portion. The mixture was filtered, and the almost colorless filtrate was shaken with charcoal and filtered again. On standing overnight at room temperature a small amount of theophylline was deposited. The filtrate from this was poured into 2 liters of ligroin (3040’) and the whole thoroughly stirred. The fluffy white precipit.ate was allowed to settle and then washed twice by decantation with ligroin. The precipitate was collected and dried. It has no definite melting point. It shrinks at about 60”. This is characteristic of similar theophylline derivatives.

1t.s specific rotation is

[cy]; = -0.10” x 100

2 X 0.136 = -36.7” (in methyl alcohol)


4.700 mg. substance: 8.730 COz, 2.380 Hz0 mg. mg. 4.565 “ “ : 0.504 cc. N (772 24”) mm.,

CIJLO&C Calculated. C 50.94, H 5.66, N 13.2

424.1 Found. “ 50.65, “ 5.66, “ 12.9

Preparation of i,bMethylorthoacetyl-3-AcetyL$-Methylrhamno- pyranose (XI)-The orthoacetate was prepared according to the method of Haworth. 1 gm. of bromodiacetyl-monomethylrhamnopyranose was dissolved in dry methyl alcohol containing 1 cc. of quinoline, and the solution allowed to stand at room temperature for 90 minutes. 50 cc. of chloroform were then added and the quinoline was removed by shaking with dilute sulfuric acid. The chloroform extract was then washed with sodium bicarbonate and finally with water and then dried over anhydrous sodium sulfate. The chloroform was removed by means of suction, and the residual colorless oil was kept under a high vacuum overnight. ni4 = 1.4540.

7 Ha\\orth, W. N., J. (‘hwr~ Sk., 2862 (1931).


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[al:: = ‘:‘“,“;;5? = +29.57” (in methyl alcohol)


4.295 mg. substance: 8.235 mg. CO,, 2.850 mg. H,O 6.740 “ “ : 11.185 “ AgI 269.0 “ “ required 14.0 cc. 0.1 N NaOH

&HSOO1. Calculated. C 52.1, H 7.3, OCH, 22.46 276.1 Found. “ 52.29, “ 7.42, “ 21.90

Calculated. COCH, 15.6 for one acetyl group I‘ ‘I 31.2 “ two I( groups

Found. ‘I 22.4

The substance was therefore a mixture of the glycoside and the orthoacetate and no effort was made to separate them.


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P. A. Levene and Irving E. MuskatSIDE

ACETONE-METHYLRHAMNOPYRANOPROPERTIES OF

THE STRUCTURE AND THE

1934, 105:431-442.J. Biol. Chem.

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