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Vol. 80
including diethyl 2,3-butadienylmalonate and 1acetoxy-2,3-butadiene. A more complete description of the synthesis of cyclobutanes from allenes and substituted olefins and the conversion of alkylidenecyclobutanes to cyclobutenes, alkylidenecyclobutenes and other dienes will be published shortly.
H , 7.40, 7.6; N, 11.0, 11.5 (Dumas); 11.8, 11.8 (Van Slyke); neut. eq., 121.5; mol. wt. 468, 444 (Rast); 427, 490 (SigneT). Kanamycin gives positive Molisch, ninhydnn and Elson-Morgan tests and negative reducing sugar, Sakaguchi and maltol tests. Treatment with 40% sulfuric acid for 100 min. a t 100" yields a product with an ultraviolet spectrum identical t o that obtained from a CONTRIBUTION No. 454 FROM THE CENTRAL RESEARCH DEPARTMENT H. N. CRIPPS pentose under similar conditions. E. I. DU PONT DE NEMOURS AND Co. J. K. WILLIAMS Treatment of kanamycin base with acetic anWILMINGTON, DELAWARE W. H. SHARKEY hydride and methanol yields tetra-N-acetylkanaRECEIVED DECEMBER 19, 1957 which was recrystallized from aqueous methanol, m.p. 250-255' dec. Anal. Calcd. for KANAMYCIN. I. CHARACTERIZATION AND ACID CzeH44N4016: C, 47.9; H, 6.8; N, 8.4. Found: HYDROLYSIS STUDIES C, 48.5; H, 6.90; N, 8.3. Kanamycin picrate Sir: was prepared by treating kanamycin base in water Kanamycin, discovered by Umezawa and co- with picric acid. The product was recrystallized workers, l is a water-soluble basic antibiotic active from boiling water, m.p. 225-230' with decomposion Mycobacteria, Gram-positive and Gram-nega- tion. Anal. Calcd. for C42H48N16039:C, 36.0; H, tive organisms. It has been characterized as the 3.46. Found: C, 36.2, 36.2; H, 3.62, 3.60. Kanamycin also has been characterized by the formation base, the sulfate and other derivatives.2 Kanamycin is isolated from fermentation broths of a series of Schiff bases. Treatment of kanamyby adsorption on Amberlite IRC-503 resin in the cin base in water with p-chlorobenzaldehyde in isosodium cycle and elution with aqueous hydrochlo- propyl alcohol yielded tetra-N-p-chlorobenzylidene ric acid. The eluate is neutralized, diluted and re- kanamycin, m.p. 213-216' with decomposition. O ~ 56.7; I: H, adsorbed on IRC-50 which has been regenerated Anal. Calcd. for C ~ B H & ~ ~ N ~ C, with ammonium hydroxide. The column is eluted 4.96; N, 5.75; mol. wt., 974. Found: C, 56.94, with 0.2 N NH40H, the eluate concentrated in 56.74; H, 5.02, 4.73; N, 5.94, 5.73; mol. wt. 971 vacuo to approximately 50-100 mg./ml. of kana- (Signer). In a similar fashion we have prepared mycin activity, diluted with 0.8-1 volume of the tetra-N-veratrylidene kanamycin, m.p. 173methanol and adjusted with H&04 to p H 8.0-8.2. 175'. Anal. Calcd. for CuH~N4019:C, 60.22; H, Kanamycin sulfate crystallizes slowly in small ir- 6.32; N, 5.20. Found: C, 60.35; H, 6.28; N, 5,14. Tetra-N-salicylidene kanamycin melted a t regular pale yellow prismatic crystals, The crystalline kanamycin sulfate may be puri- 272-274' with decomposition. Anal. Calcd. for fied by repeated recrystallization from methanol- CaeHszNaOlb: C, 61.33; H, 5.77. Found: C, water a t p H 7.8-8.2 to give white irregular pris- 61.25; H, 5.75. Tetra-N-p-methoxybenzylidene matic crystals. This preparation contains ad- kanamycin melted a t 193-196' with decomposiO I62.7; ~: H, sorbed moisture which is removed with difficulty. tion. Anal. Calcd. for C E , ~ H ~ O N ~C, Samples for analytical determinations were dried 6.27. Found: C, 62.6; H, 6.66. Paper chromatography of kanamycin preparato constant weight a t 170" in vacuo. Anal. Calcd. for C18H3~N4OI1.H1SO4:C, 37.13; H, 6.58; N, tions revealed a second antibiotic, designated kana9.62; S, 5.50; Sod-, 16.6; neut. eq., 145.6; mol. mycin B. The two antibiotics are best separated wt., 600.6 (monohydrate). Found: C, 37.3, 37.4, in Peterson's n-butanol-~ater-27~ p-toluenesul37.3; H, 6.8, 6.6, 6.3; N, 9.3, 9.6; S, 5.5; SOr-, fonic acid system6 on Schleicher and Schuell 589 16.8; neut. eq., 146.6; mol. wt. 604 (X-ray on un- Blue Ribbon or Whatman No. 1 papers. I n t h i s dried m a t e ~ i a l ) . ~ The formula containing two less system with S&S 589 Blue Paper, kanamycin has hydrogen atoms, although improbable, is not ex- an R p of about 0.35 and kanamycin B has an R p of cluded by analyses on this and other derivatives. about 0.6. The presence of impurities or contamiThe sulfate is soluble in water, insoluble in the nating salts interferes markedly with the paper common alcohols and non-polar solvents. It shows chromatography of the kanamycins in this system. no melting point, decomposing over a wide range The infrared spectra of kanamycin and kanamyabove 250". It was converted to kanamycin base cin B are similar. Each is typical of a polyhydroxy, by treatment of an aqueous solution with a strongly polyamino compound. No carbonyl or carbonbasic anion-exchange resin, concentration and carbon double bond adsorption is evident. crystallization with methanol-ethanol mixture, Kanamycin is remarkably resistant t o acid and [ c u I z 4 ~4-146 (c 1, 0.1 N , H2S04). Anal. Calcd. for alkaline hydrolysis. Treatment of kanamycin C I ~ H ~ ~ NC, ~O 44.6; ~ I : H , 7.5; N, 11.6; neut. eq., with methanolic hydrogen chloride under condi181.1; mol. wt., 484.5. Found: C, 44.7, 45.0; tions which hydrolyze neomycin B and C to neomycin A (neamine)+* yielded unchanged starting (1) T . Takeuchi. T. Hikljl, K. Nitta, S. Yamazaki, S. Abe. H. Takayama and H. Umezawa, J . Anlrbiolics. 810, 107 (1957). material. Refluxing kanamycin base in 6 N HCI
( 2 ) (a) H. Umezawa, M. Ueda, K. Maeda, K. Yagishita, S. Kondo. Y. Okami, R. Utahara, Y. Osato, K. Nitta and T. Takeuchi, ibid., AlO, 181 (1957); (b) K. Maeda, M. Ueda, K. Yaplshita, S. Kawaji, S. Kondo, M. Murase, T . Takeuchi, Y . Okami and a.Umezawa, ibid., AlO, 228 (1957). (3) A product of the Rohm and Haas Co.
(6) D. H. Peterson and L. M. Reinecke, THIS JOURNAL, 72, 3598 (1960). (5) J. D. Dutcher, N . Hosansky, M. N. Donin and 0. Wintersteiner. ;bid., VS, 1384 (1951). (7) B. E. Leach and C.M. Teeters, ibid., 13, 2794 (1951).
(4) The X-ray determination was run at the Massachusetts Institute of Technology, through the courtesy of Dr. David P. Shoemaker.
(8) R. L. Peck, C. E. Hoffhine, Jr., P. Gale and 76, 1018 (1953).
K. Folkerr. a i d . ,
Feb. 5, 1958
COMMUNICATIONS TO THE EDITOR
for 45 min. completely destroyed the biological activity. After decolorizing, the solution was taken to a small volume and methanol and ethanol were added, yielding crystalline 1,3-diamino-4,5,6trihydroxycyclohexane dihydrochloride (desoxystreptamine) ? Anal. Calcd. for CsH14N203.2HCl: C, 30.66; H, 6.86; N, 11.93; C1, 30.17; neut. eq., 117.5. Found: C, 30.76; H, 6.99; N, 11.36; C1, 29.6; neut. eq., 118.7. The compound showed no optical activity in acid or alkaline aqueous solution. The dihydrochloride was converted to desoxystreptamine by dissol_ving in water, treating with Amberlite I R 410 (OH),3 concentrating, and crystallizing with ethanol. Anal. Calcd. for C6H14N203: C, 44.4; H, 8.70. Found: C, 44.66; H, 8.86. A portion was converted to the dihydrobromide by crystallization from dilute aqueous HBr with methanol. It gave an infrared spectrum identical with that of an authentic sample of 1,3-diamino4,5,6-trihydroxycyclohexanedihydrobromide isolated from neomycin. lo The desoxystreptamine was additionally characterized by acetylation in pyridine and acetic anhydride to give pentaacetyl1,3-diamino-4,5,6-trihydroxycyclohexane. Anal. Calcd. for C16H24N208: C, 51.6; H, 6.48; N, 7.53. Found: C, 51.7; H, 6.98; N, 7.30. Selective de0-acetylation with Amberlite I R 410 (OR)%in water yielded N, N ’-diacetyl-1,3-diamino-4,5,6-trihydroxycyclohexane. Anal. Calcd. for CloHlsNz0 6 : C, 48.77; H, 7.37. Found: C, 48.82; H, 7.73. Other products of the acid hydrolysis shown to be present by paper chromatography include two ninhydrin-positive reducing materials, accounting for the major portion of the material left after removal of desoxystreptamine, and several minor components. These data allow speculation that kanamycin is a trisaccharide-like molecule, composed of two aminosugar moieties glycosidically linked to desoxystreptamine.
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Consequently, i t appears highly probable that the cation of I should have an octahedral configurationj3s4 which leads t o the conclusion that i t should be asymmetric. A resolution of racemic [Si(acac),]+ into its optical enantiomers6 would offer conclusive evidence for its octahedral configuration. Optical Resolution Studies.-Racemic [Si(acac)3]Cl*HClwas prepared by a modification of the method of Dilthey6 and its resolution was attempted by treatment with sodium (-)-dibenzoyld-tartrate,?+ according to the equationlo 2D,L-[Si(acac)a]Cl.HCl-I- d-(-)-Na&HlzOg + ~-[Si(acac)a]d-( -)-HC18H12O8 ~-[Si(acac)~]Cl.HCl -I2NaCI (1)
+
The diastereoisomer L- [Si(acac)s]d-(-)-HC18HI208is less soluble than the other diastereoisomer and precipitates from solution. For L- [Si(acac)r]d-( -)-HC1~H1~08,a 0.008870 solution showed sobs to be -0.068 + 0.003’; [ r u I z 3 ~-773’. The optical rotation of the diastereoisomer decreased (became more positive) with time as the asymmetric silicon complex ion racemized, and, after six hours, the observed rotation decayed to a constant value ( -O.OOSO) due to the asymmetric monohydrogen dibenzoyl-d-(-)tartrate anion. I n order to replace the resolving anion with chloride, the diastereoisomer was treated with a n anion-exchange resin in the chloride form (Dowex 1-X8), which gave ~-[Si(acac)~]CI.HCl.A 0.0311% solution of this enantiomer showed CUobs t o be -0.300 The observed rotation de0.003’; [ r u I 2 3 ~-965’. creased (became more positive) as the compound racemized, and decayed t o zero degrees in six hours with a half-life of one hour. By extrapolation, the specific rotation, [cY]’~Da t zero time is found to be -1,300°. I n order to determine whether the anion-exchange resin removed the negative rotating monohydrogen (- )-dibenzoyl-d-tartrate anion (9) D. A. Kuehl, Jr., M. N. Bishop and K. Folkers, THISJOURNAL, 73, 881(1951). quantitatively from the diastereoisomer, an experi(10) Supplied by Dr. H. E. Carter, University of Illinois. ment was performed in which a sample of soM. J. CRON dium monohydrogen (-)-dibenzoyl-D-tartrate was D. L.JOHNSON treated with resin under similar conditions; the RESEARCH DEPT. F. M. PALERMITI BRISTOL LABORATORIES, INC. Y. PERRON filtrate showed no optical rotation, indicating that SYRACUSE I, NEW YORK H. D. TAYLOR essentially complete removal of the asymmetric D. F. WHITEHEAD anion had been achieved. I. R. HOOPER The optical rotation of the filtrate from reaction RECEIVED JANUARY 2, 1958 1 was found to be strongly dextrorotatory. It was treated with the chloride form of an anion-exchange resin (Dowex 1-XS) in order to remove the THE RESOLUTION OF A HEXACOVALENT SILICON monohydrogen (- ))-dibenzoyl-D-tartrateanion, and (IV) COMPLEX its rotation was measured. A 0.1 164y0 solution Sir: showed a 0 b o = 0.790 f 0.03’; [ruIz3D +679O. The possibility of six-coordinate silicon(1V) has The observed rotation of this solution of the dexbeen discussed by Stone and Seyferth‘ who de(3) A. Werner, Ber., 47,3087(1914). scribed the availability of d-orbitals for bonding (4) J. C. Bailar, Jr., Ed., “Chemistry of the Co(lrdination Comin silicon(1V) compounds. West2 described the in- pounds,” Reinhold Publishing Co., New York, N. Y., 1958. PP. 274 frared spectra of [Si(acac)3]Cl.HCI (I) and other ff. (5) acac acetylacetonate anion, CrH?Ol-. This cation Is also P-diketones of silicon(IV), and from a comparison tris-(2,4-pentanediono)-silicon(IV)cation. of these spectra with that of free acetylacetone named (8) W. Dilthey, Bn.,86, 932,1595 (1903). concluded that the ligands of I behave as bidentate (7) S.Kirschner, Dissertation, University of Illinois, 1954,p. 43. (8) C. L. Butler and L. H. Cretcher, TEIS JOURNAL, 66, 2805 chelating agents.
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(1) F. G. A. Stone and D. Seyferth, J . Inorp. N U C ~ CChem., Q ~ 1, 112 (1955). (2) R. West, Abstr. of Papers, 132nd Meeting, Am. Chem. Soc.. 11-N(1957),THISJOURNAL, in press.
(1933). (9) F.Zetzsche and M. Hubacher, Rclu. Chim. A c f a ,9,291(1928). (10) “n” and “L” indicate dextrorotatory and levorotatory complex Ions, respectively; the “d-( -)-” prefix for dibenzoyltartrate indicates a negative rotating anion derived from dextrorotatory tartaric acid.
