NOTES
Kovember, 1963
801
of benzene and chloroform as shown in Table I. Final stripping p-Aminoanisole-C1*.-A solution of p-nitroanisole-C14 (223 mg., was accomplished with ethanol and methanol. 1.45 mmoles) in 95% ethanol (15 ml.) containing platinum oxide catalyst (20 mg.) was hydrogenated at slightly above atmospheric pressure a t 25". After the reduction was complete (1 hr.), the TABLEI solution was filtered under an atmosphere of CO2 into a flask ELLERT COMPOSITIO~ FOR ALLVIPUA C O L L WCHROMATOGRAPHY ~ containing 6.0 N sulfuric acid (1.0 ml.). The catalyst and reaction flask were rinsed with ethanol and the rinsings filtered and OF DIUITRODIPHENYL ETHER DERIVATIVE transferred to the main solution. Evaporation of the ethanol Chloroafforded crystalline p-aminoanisole-C'4 hydrogen sulfate. The Volume, Benzene. form, Radioproduct was freed from last trares of ethanol by three evaporaFraction ml ml. ml actiritya tions of its suspensions in anhydrous benzene (10 ml. each). 50 50 0 Trace IA p-Methoxyphenol-C'*.-The light tan p-aminoanisole-C1* IIA 100 90 10 Trace hydrogen sulfate (1.45 mmoles, assume 1007, reduction) was 100 80 20 Trace IIIA suspended in 3 M sulfuric acid (2.5 ml.) and diazotized a t 0 to '5 with sodium nitrite (102 mg., 1.48 mmoles) in water (1.2 nil.). 350 175 1i 5 Trace IVAa The diazonium solution was stirred a t 0 to 5" for 15 min., then 100 50 50 High IYAb the excess nitrite was destroyed with urea. The diazonium salt 50 25 25 High VA w-as hydrolyzed by boiling it with 6 S sulfuric acid (50 ml.) 300 0 300 High VIA for 30 min. The reaction mixture was cooled and extracted High VIIA 200 Absolute ethanol with four 20-ml. portions of peroxide-free ether. After the ex1111.4 200 .4bsolutr methanol High traction, the dissolved ether was evpelled from the aqueous phase by careful heating and the aqueous mixture refluxed again h h q u n t s nere counted in the Packard Tri-Carb liquid scintilfor 30 min. This process of boiling for 30 min., ether extraction, lation counter. expulsion of dissolved ether from reaction mixture, and refluxing the hydrolysate again was rarried out a total of eight times. Fraction IYAb contained the bright yellow band from the The combined yellow ether extract was concentrated to about column characteristic of the dinitrodiphenyl ether derivative. 30 ml., and the resulting ether solution was washed with three Fractions T'-4, T'I.4, VIIA, and VIIIA each afforded on evapora5-ml. portions of saturated sodium chloride solution. The prodtion small amounts of uncrystallizable oils and gums ranging uct was then extracted with four 10-ml. portions of S sodium in color from yellow to brown. These radioactive by-products hydroxide. The sodium hydroxide extracts were combined and from the condensation reaction were not investigated. acidified to pH 1 with concentrated hydrochloric acid, saturated Evaporation of solvent from fraction IVAb afforded the diwith sodium chloride, and extracted with ten 10-ml. portions of nitrodiphenyl ether derivative (215 mg.) which was recrystallized ether. The ether was evaporated and the residual oil rendered from t-butyl alcohol ( 1 in1 ), washed with two 0 1-ml. portions of anhydrous by two evaporations of its solutions in dry benzene t-butyl alcohol, and dried 7n z'ucuo over phosphorus pentoxide. (10 ml. each). The crude red-brown semisolid p-methoxyThe light yellow ?;-acetyl-3- [4-(4-methoxyphenoxy-C14)-3,5-diniphennl-CI4 amounted to l i 7 mg. (98Yc based on p-nitroanisole-C14 trophenyll- alanine ethyl ester (158 mg , 155; based on phenolor 627,12 based on phenol-C14). A melting point determination C") melted a t 98-101" (softens %'), lit.*m p 109-110". was not practical, lit.'3 m p. 53". h paper strip (Whatman KO. 1) N-Acetyl-3- [4-(4-methoxyphenoxy- W) -3,5-diaminophenyl] chromatogram was developed by ascending technique in benzeneL-alanine Ethyl Ester.-A solution of N-acetrl-3-[4-(4methpropionic acid-water (100:iS: 5 ) . A single radioactive peak was oxyphenoxy-C1*)-3,5-dinitrophenyl]-~-alanine ethyl ester ( 112 obtained which corresponded to authentic nonradioactive p mg., 0 25 mmole) in 95% ethanol (10 ml ) containing platmethoxyphenol a t Rr 0.71. On spraying the chromatogram with inum oxide catalyst (10 mg.) was hydrogenated a t slightly diazotized sulfanilic acid, a single spot developed which correabove atmospheric pressure a t room temperature (25'). During sponded with the radioactive peak. the reduction, the system was protected from excessive light. N-Acetyl-3- [4-(4-rnethoxypheno~y-C'~)-3,5-dinitrophenyI] After the reduction was complete (about 2 hr.), the mixture L-alanine Ethyl Ester.-The procedure of Chalmers, et al.,z was cooled in ice, the flask was evacuated, and glacial acetic acid was followed with modification of the amounts of reactants. (0.3 ml.) mas added with stirring. The flask was flushed mFith From lX-acetyl-3,5-dinitro-~-tyrosineethyl ester2 (470 mg., 1.38 CO2, and all subsequent manipulations were conducted in an mmoles), p-toluenesulfonyl chloride (263 mg., 1.38 mmoles), atmosphere of CO?. The reaction mixture was filtered, and the anhydrous pyridine (1 ml.), and p-methoxyphenol-C14 (177 mg., solvent was evaporated. The residual pale yellow oil was ren1.42 mmoles) dissolved in pyridine (1.6 ml.) there was obtained dered anhydrous by evaporating its solution in absolute ethanol the yellow-brown crude S-acetyI-3-[4-(4-methoxypheno~y-C1~)- (25 ml.) and finally rendered alcohol-free by t x o evaporations of 3,5-dinitrophenyl]-~-alanine ethyl ester (405 mg., 66% based on its solutinns in anhydrous benzene (25 ml. each). The resulting tyrosine derivative) which melted a t 93-100" (softened a t 68"). yellow viscous oil of S-acetyl-3-[4-( 4-methoryptienoxy-C1*)-3,5The unchanged p-methoxypl1enol-C1~ was recovered from the diaminophenyll-L-alanine ethyl ester diacetate was used without sodium hydroxide extracts by acidification t o p H 1 with conpurification. centrated hydrochloric acid, saturation with sodium chloride, and N-Acetyl-3- [4-(4-methoxyphenoxy-C'J) -3,5-diiodophenyl] extraction with ten 10-ml. portions of ether. The combined L-alanine Ethyl Ester.--A slightly modified'* procedure of ether extracts were extracted with three 10-ml. portions of N Chalmers, et U L , ~was followed with proportional reduction sodium bicarbonate. The ether solution was filtered through a in the scale of materials throughout. From the crude N-acetylcotton plug and the ether evaporated. The residual oil was 3- 14-( 4-metlioxypheno~y-C1~)-3,5-diaminophenyl] -L-alanine ethyl rendered anhydrous by two evaporations of its solutions in anester diacetate (0.25 mmole) obtained from the preceding reduchydrous benzene (10 ml. each). The recovered p-methoxytion, there was obtained light buff colored prisms of ?r'-acetyl-3phenol-C1* (33.5 mg.) remained as a yellow oil. By carrying out [~-(~-methoxyphenoxy-C~4)-3,5-diiodophenyl]-talanine ethyl a similar condensation on the recovered p-methoxyphenol-C'*, ester (109 mg., 727, based on the dinitrodiplienvl ether derivathere was obtained a second crop of the dinitrodiphenyl ether tive) uhich melted a t 136-13T0, l i t 2m p. 143-144'. derivative (85 mg.), n1.p. 89-98' (softened a t 83"). The two 3,5-Diiodo-~-thyronine-C~*.-Theprocedure of Chalmers, crops of crude condensation product were combined (490 mg. j, et a1.,2 was followed with proportional reduction in the scale of recrystallized from f-butyl alcohol (1.5 ml.), and dried over phosreactants throughout. From X-acetyl-3- [4-(4-methoxyphenphorus pentoxide zn mcuo. oxy-C14)-3,.5-diiodophenvl]-~-alanine ethyl ester (97 mg., 0.16 The recrystallized S-acetyl-3-[4-(4-methoxypheno~y-C~~)-3,5-mmole), hydriodic acid (freshly distilled 57%, 0.25 ml.), and dinitrophenyl]-~-alanine ethyl ester (280 mg., m.p. 88-98') glacial acetic acid (0.25 ml.)) there was obtained crude white was dissolved in anhydrous benzene ( 3 ml.) and chromatographed 3,5-diodo-~-thyronine-C1* (45 mg., 5470 based on 3,5-diiodo on a column of Woelm alumina (neutral, grade 1 ) measuring derivative) which melted a t 244' dec., lit.2 n1.p. 2.55' dec. It was 100 X 31 mm. The elution was carried out with a mixture used nithout purification.
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-
-
(12) Probably not a valid representation of degree of conversion because of the impure nature of the product. (13) H. Hlasiwetz and J. Habermann, Ann., 177, 339 (1875).
(14) After the finely pulverized sodium nitrite was added t o concentrated sulfuric acid, the mixture was warmed t o 70" until solution was complete. I t was then cooled t o On and cautiously diluted with glacial acetic acid.
