The synthesis and testing of some alkyl substituted thyroxine analogs

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Content T H E SYNTHESIS A N D T E ST IN G . O F S O M E A L K Y L SUBSTITUTED T H Y R O X IN E A N A L O G S by John Douglas Arterberry A Thesis Presented to the F A C U L T Y O F T H E G R A D U A T E S C H O O L UNIVERSITY O F S O U T H E R N C A LIFO R N IA In. P a rtia l F u lfillm en t of the Requirements fo r the Degree M A S T E R O F SCIENCE (Chemistry) August 1955 UMI Number: EP41608 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion. Dissertation Publishing UMI EP41608 Published by ProQuest LLC (2014). Copyright in the Dissertation held by the Author. Microform Edition © ProQuest LLC. All rights reserved. This work is protected against unauthorized copying under Title 17, United S tates Code ProQ uest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, Ml 48106 -1346 UNIVERSITY O F S O U T H E R N CA LIFORNIA G R A D U A T E S C H O O L U N IV E R S IT Y P A Rk L O S A N G E L E S 7 This thesis, written by . . . . . . ...................................................... under the guidance of h.i.s.— Faculty Committee, and approved by all its members, has been pre sented to and accepted by the Faculty of the Graduate School, in partial fulfillment of the requirements for the degree of M a ste r o f S c ie n c e , Dean j ? s y Faculty Committee F . Kharasch Chairman ACKNOW LEDGEM ENT The author wishes to thank Professor Norman Kharasch for h is many h elp fu l suggestions and valuable guidance in the stu d ies presented herein. G rateful acknowledgement i s also made to the United S tates Public Health Service for support of these s tu d ie s . TABIE OF CONTENTS) C H A PT E R P A G E I . INTRODUCTION...................................................... .... 1 I I . A. RA TIO N A LE FO R A LK Y L SUBSTITUTED A N A L O G S O F 7 T H Y R O X IN E ............................................................................. I I I . SYNTH ETIC ASPECTS, 10 The S yn th esis o f S everal New Thyroxine Analogs . ' 10 Reagents and S olven ts ............................................................ 22 Experim ental .................................................. 2i* ii-Hydroxyhydrocinnamic Acid .. .. .. .. . 2it E thyl 3* 5-dinitro-ii-hydroxyhydrocinnam ate . . 26 E thyl 3 i 5 -d in itr o -it-(3 1 -m ethyl-it1 -m ethoxy- 27 phenoxy)-hydrocinnamate . . . . . ................... E thyl 3} 5 -d in itr o -lt-(2 1 -m ethyl-it’ -m ethoxy- 28 phenoxy)-hydro cinnamate ..................................... Ethyl 3 » 5>-dinitro-it~(2 ’ ^3*-dim eth yl-it1- 29 hydroxyphenoxy)-hydrocinnamate ....................... . E thyl 3, 5 -d in itr o -it-(3 ! -dime thy l - V - 30 methoxyphenoxy)-hydrocinnamate . . . . . . . E thyl 3 3 5 - d in it r o -it - (2 '36*-d im eth yl-lil - 31 methoxyphenoxy) -hydrocinnamate ............................ E thyl 3 3 5>-dinitro-it-(2 ’ ^ ’ -d im eth y l-lt'- 32 methoxyphenoxy )-hydrocinnamate ...................... • E thyl 3> £ -d in itr o -li-(2 * , 3 ’ jf> S6‘- t e t r a - 33 methyl-ii. * -hydroxyphenoxy)-hydrocinnamate . . . 2 -Me thyl-ii-m e th oxy-5-isop rop ylp h en ol . . . . 3k E thyl 3 , 5 -d in itr o -it-(2 1 -m ethyl-5 * -iso p r o p y l- it' -hydroxyphenoxy) -hydro cinnamate ........................ 3h l i CHAPTER PAGE E th yl 3 , 5 -d in itr o -li- (2:* , 5 1 -d i-t-b u ty l-U ! - hydroxyphenoxy)-hydrocinnamate . ................ 35 E th yl 3 ,5 -d in itr o -U -(2 ' ^ '-d im e th o x y -V - hydroxyphenbxy)-h y d r o c in n a m a te ............................... . . 3-5 E th yl 3 ,5 -d in itr o -lj- (2 1} 5 1 -dim ethoxy-li * - hydroxyphenoxy)-hydrocinnam ate......................................... 3 * 5 E th yl 3 ,5 -d in itr o -ii-(2 ’ j^ ’-d ieth oxy-U ’- hydroxyphenoxy)-hydro c in n a m a t e ......................................... 37- E thyl 3 j5 -d in itr o -ii- ( 2 1 ,5" -di-n-propoxy-1*' - hydroxyphenoxy)-hydrocinnamate.............................................. 38 3 55 -D iio d o -i;-(3 1 -m ethyl-i;' -hydroxyphenoxy) - hydrocinnamic Acid ................... .......... 38 3 ,5>-Diiodo-!i-(2 ’ , 5 ’ -dim ethoxy-U' -hydroxyphenoxy)- hydrocinnamic Acid" - .......................................................... 39 A n a ly tic a l Data ....................... . . . . . . . . . . . IV. T H E FEPHIC C H L O R ID E OXIDATION O F 2 a 6-DIALKYL PH E N O L S ....................................' ................................ W - Experim ental . . . . .. .. . .................................... . bS 3 s 5 *31 >51 -T etra -t-b u ty l-l* , U' -diphenoquinone . . . 3 j5 j3 ' , 5 l -T e tr a -t-b u ty l-iiJii, -diphenohydroquinone . b5 3 j5 33 1 ,5 ' -T etra -i-p ro p y l-iijli' -diphenohydroquinone. A n a ly tic a l Data ................................................. W 3 V. PRELIM INARY BIOSSAY O F A N A L O G S ................................ 1 * 9 VI. S U M M A R Y A N D C O N C LU SIO N S .......................................................... 52 BIBLIOGRAPHY . . . .................................................................................. 53 LIST OF TABLES T A B L E P A G E I . A n alytical D a t a ............................................................... . . U O I I . Infrared Absorption Spectra . ................................................. iUi III. A n alytical Data ................................................................ k8 IV. A c tiv itie s of Thyroxine Analogs as Determined by the Rana Gatesbeiana T ail-F in Assay . . . . . . f>l ! I CHAPTER I IN T R O D U C T IO N ! | In 1820, Goindet (1) demonstrated the importance o f elem ental • io d in e in the treatm ent o f io d in e d e fic ie n c y g o it e r , but i t was not i | u n t il 1895 th a t Baumann demonstrated iod in e as an e s s e n t ia l component i j o f the thyroid gland ( 2 ) . In th a t same year Oswald named the io d in e - * • • • | con tain in g c o llo id th yroglob u lin (3 ) . During the fo llo w in g decade ; other workers showed th a t the iod in e p resen t in the gland tended to ; vary in v e r se ly w ith the degree o f h yp erp lasia o f the gland (h ). By 1 9 1 3 -lli, assay methods fo r th y ro id -a c tiv e substances were developed, : and in 19ljl Kendall is o la te d c r y s ta llin e L-thyroxine ( 5 ) . In 1926, j Harington completed the proof o f stru ctu re and the n ext year in c o l- ! la b o ra tio n w ith Barger, sy n th esized D,1.-th y ro x in e, 1 -1 , ( 6 ). i In 1935a S a lte r , Lem an, and Means (7 ) reported th a t the D, i the L, and the B,L forms o f thyroxine were eq u ally a c tiv e in tre a tin g myxedematous p a t ie n t s . To the con trary, animal experim ents have ; shown the L form to be the a c tiv e form. The sim ple one-ring compound, ■ L -3 ,5 -d iio d o ty r o sin e , 1 -2 , i s a c tiv e only in dosages hundreds o f j tim es g rea ter thah th a t o f th y ro x in e, and even th is a c t iv it y has been z t ^ X i ! - iconsidered to be only a r e s u lt o f i t s a ctio n as a precursor e ith e r 2 o f the hormone i t s e l f o r th yroglob u lin ( 8 ). Mechanisms by which thyroxine may be formed from two m olecules o f L -3*5-d iiod otyrosin e have been advanced by Johnson and Tewkesbury ( 9 ) s Harington (1 0 ), and Neuberger (1 1 ). In 1937 Harington estim ated fo r ty percen t o f the o rg a n ica lly bound iod in e o f the th yroid to be in th yroxin e, and the remainder to be in L -3 ,5 -d iio d o ty ro sin e (1 2 ). Work completed w ith in the l a s t ten years has shown th a t much, but fa r from a l l , o f the m eta b o lica lly a c tiv e o r g a n ica lly bound iodine, of th yroglob u lin i s in the form o f th yroxin e. Thus, fo r se v e r a l years an iod in e ^-containing hormone, more a c tiv e than thyroxine i t s e l f , was sough t. This e f f o r t culm i nated in the discovery by Gross and P itt-R iv er s (13)) and Roche and co-workers (19) o f L-3 *3 * -triio d o th y ro n in e 1-3* The r e la tio n sh ip e x is tin g between thyroxine and triiod oth yron in e i s p resen tly uncer ta in ; however the la t t e r i s b elie v e d to be a d e r iv a tiv e o f the former ( lit ) . P reviously L -3-iod otyrosin e was shown to be a d eriv a tiv e o f fra ctio n ed th y ro g lo b u lin . T riiodothyronine has se v e r a l tim es the a c t iv it y o f th y ro x in e, both in mammalian and amphibian assay (1 3 ). : r Among the p h y sio lo g ic a l e f f e c t s o f th yroid hormone are: (a) reg u la tio n of b a sa l m etabolism , in clu d ing p r o te in , f a t , carbo- 3 \ hydrate, s a l t and w ater m etabolism , (b) a ctio n on the ra tes o f 5 growth and d iffe r e n tia tio n o f body t is s u e s , (c ) a ctio n on the neu- f romuscular and c ir c u la to r y system s, and (d) in te r a c tio n w ith the i other endocrine g la n d s, e s p e c ia lly the a n terio r hypophysis in the ' maintenance o f hom eostasis (15) • I t has been shown th a t the a ctio n | o f sy n th e tic analogs i s akin to th a t o f th yroid hormone, w ith the / , d iffe re n ce being one o f r e la tiv e potency o f the p a r tic u la r analog as 1 ! compared to thyroxine (1 6 ). C an zan elli and Rapport (18) had compared ty r o s in e , d iio d o - | ty r o sin e , th yron in e, and thyroxine w ith re sp e c t to th e ir a c t i v it i e s I ! on b a sa l m etabolic r a t e . They found the in crea sin g order o f a c t iv it y | | to be Tyrosine D iio d o ty ro sin e Thyronine Thyroxine. From t h is data Kendall concluded th a t the e f f e c t o f thyroxine was q u a lita tiv e ly unique, a n o tio n p resen tly untenable in view of the i la rg e number o f thyrom im etically a c tiv e sy n th e tic compounds. When Niemann f i r s t reported on stu d ie s in the thyroxine f i e l d , in 19iH , i t had already been esta b lish e d th a t hormonal potency was not a unique fu n c tio n o f the alanine sid e ch ain . He looked upon the sid e chain as m erely conferring p o la r ity upon the m olecule by v ir tu e o f the term inal carboxyl and/or amino group. He fu rth er p red icted th a t the sid e chain could be moved to the or the 6 p o s itio n , from i t s normal sp o t a t the 1 p o s itio n , and th a t th ese p o s itio n a l isomers, would show a c t iv it y (2 0 ). Niemann has fu rth er emphasized th a t the b a sic str u c tu r a l nucleus i s th a t as proposed by Harington (21) and by F rieden and W inzler (1 7 ), and which S a lte r has c a lle d the b a sic pharmacogen. Niemann and h is co-workers (2 0 , 22, 23) su ggest th a t I-it may, in v iv o , undergo oxid ation r e v e r sib ly to 1 -5 , p assin g through a semi-quinone in term ed ia te. The ortho analog o f th y ro x in e, u n lik e the meta an alog, i s capable o f forming a quinoid str u c tu r e , a t le a s t in th eory, as p o stu la ted by Niemann and Mead (2 2 ). The ortho an alog, in the h y p o th etica l o x id ized form, i s represented by 1 -6 . The meta analog, 1 -7 , was sy n th esized by Niemann and Redemann (2 3 ), and i t was found in a c tiv e on the b a sa l m etabolic ra te s o f r a ts ; up to a dosage o f 500 m g./k g. by Boyer (2l±). Using the same t e s t , | Boyer found the ortho analog to have 2-h% a c t iv it y , as compared to 1 . • | thyroxine (2li) M < V o U ^ - C c o / / I - / / \ V - o - / V ' 1 ' X i i A new meta analog, 1 -8 , has r e c e n tly been reported by Bruice (25) j to have s lig h t a c t iv it y in the tadpole metamorphosis t e s t . However, ) _ j Kharasch has pointed out (26) th a t th is observed a c t iv it y may be due j to a tra ce o f the extrem ely a c tiv e an alog, 1 -9 , reported by B ru ice, | W inzler, and Khatasch (2 7 ). Approximately 0.015# o f 1-9 would s u f f ic e 5 to giv e the a c t iv it y reported fo r 1-8* This compound, along w ith others reported by th ese w orkers, was assayed u sin g the Rana C atesbeiana tadpole metamorphosis t e s t which i s exceed in gly sen s it i v e to analogs o f desam inothyronine, the s o -c a lle d propionic acid analogs (2 7 ). The only non-halogen thyroxine analog thus fa r reported as having appreciable a c t iv it y i s 1-10 (2 7 ), which was 2 .7 # a c tiv e in the tadpole a ssa y , compared to th y ro x in e. These workers a lso pre pared 1 -1 1 , and 1 -1 2 , the former showing an a c t iv it y o f 0 .2 # , the la t t e r 280#. i - X X 1 toori i off X dtf 3 6^3 : ■ cM* c4l* cH v- tD -tC D } ) I Analogs in which the p henolic group has been blocked by m ethylation show dim inished a c t iv it y (28, 29 , 1*9). Blocking the fr e e carboxyl group by e s t e r if ic a t io n , or the amino group by a c e t- y la tio n , produces a sm aller red u ction in a c t iv it y (2 9 ). The only a c tiv e analog rep orted thus f a r , in which the diphenyl eth er linkage CoDb -CCQh I ! 6 j i s v a r ie d , i s th a t reported by Harington in 191*8, in which an | atom o f su lfu r rep laced the eth er oxygen atom (JO), s t i l l g iv in g a i j harmone w ith 20$ a c t iv it y . The high a c t iv it y o f tr iio d o th y r o n in e , 1 -3 , in both mammals, and in amphibia (2 7 ), and the high a c t iv it y o f 3 s , 5 ’“dim ethyl- 3 ,5 -d iio d o th y ro n in e, 1 -1 2 , i s su r p r isin g . This l a t t e r su b stan ce, in the tadpole metamorphosis t e s t , showed an a c t iv it y o f 280$ compared to D ,L-thyroxine (2 ? ). Analogs o f stru ctu re 1-13 have g rea ter a c t iv it y than comparable analogs o f stru ctu re I -11*, which in turn are more a c tiv e than those o f stru ctu re I - l 5 . * "A /T~\ _ v //_ /7~\ \ ; As summarized by Bruice (1*9), the decreasing order o f a c t iv it y fo r X i s I ABrXN02^ H, and the decreasing order o f a c t iv it y fo r X1 ! i s CH^ 1 ^ Br^ O l^ . The ex a ct p o sitio n o f CH^- i s un- : c e r ta in . The ph en olic rin g appears to be o f le s s importance to th y- i ; romimetie a c t iv it y than the rin g bearing the sid e ch a in , sin c e var- | yin g the 3 1 and 5* su b stitu e n ts has a le s s d r a stic e f f e c t upon the ! | a c t iv it y o f the analog than does varying the 3 and $ p o sitio n sub s titu e n ts . Although the alan in e sid e chain appears to be more s p e c if ic ; fo r mammals, the ob servation th a t high tadpole a c t iv it y i s endowed i : by the prop ion ic a cid sid e chain has in creased the range and accuracy ! i | of te s tin g stru ctu res o f h ith e rto low a c t iv it y . CHAPTER I I ] A R A TIO NALE F O R A L K Y L SUBSTITUTED ; j A N A L O G S O F T H Y R O X IN E ■ i i As sta te d in the preceding Chapter, the high thyromimetic j a c t iv it y reported fo r 1-12 and fo r substances o f the g en eral stru ctu re j I-1 3 i s not only cu r io u s, but i s deserving o f fu rth er stu d y . By j analogy, i t may be p red icted th a t substances o f the gen eral stru ctu re | I I - l would a lso show high a c t iv it y , as would stru ctu res of the form ‘ I I - 2 . Here, R could be e ith e r alanine or propionic a c id , X could be n itr o or halogen, X1 could be n itr o or halogen, and R11 could be e ith e r a lk y l or alk oxy. A second, a th ir d , or even a fou rth R” group may be added. The e f f e c t on a c t iv it y due to 2 * ,6 ’ su b stitu e n ts i s o f fu rth er in te r e s t in so fa r as Niemann and MeCasland syn th esized I I -3 (3l ) , which was la te r reported to be in a c tiv e . A second and h ig h ly p r a c tic a l co n sid era tio n favorin g the study o f a lk y l and alkoxy analogs i s th e ir r e la t iv e a c c e s s ib ilit y through the Glaxo m o d ifica tio n o f the Ullman-Nadai r ea c tio n (3 2 ), allow ing compounds o f stru ctu re II-L to be coupled w ith compounds of stru ctu re II-l? to give diphenyl eth ers o f stru ctu re I I - 6 . 8 «/H C C H - ft1 " * 4 l\ i H CV - a/HCocH^ C r tfC N O c.-o cM ^ c# s I I The prop ion ic acid sid e chain i s considered p refera b le to the alan in e sid e ch ain , because o f i t s g rea ter a c t iv it y in the tad p ole metamorphosis a ssa y , as shown by B ru ice, W inzler and Kharasch s (2 7 ). For the case o f th yroxin e, 1 -1 , as compared w ith desamino- th y ro x in e, 1 -9 , the la t t e r i s 130 tim es as a ctiv e as the former (2 7 ). Thus i t i s hoped th a t the sim ple d in itro d ip h en y leth er a n a lo g s, such as I I - 6 , w i l l be in creased in a c t iv it y to such an e x ten t th a t they may be compared d ir e c tly w ith a thyroxine standard a t comparable ranges o f c o n cetra tio n . S ince the "unprime1 1 or sid e -c h a in rin g i s h eld co n sta n t, the expected d iffe r e n c e s in a c t iv it y may be a ttr ib u te d to the a lk y l or alkoxy su b stitu e n ts on the "prime1 1 r in g . TABLE I I I A N A LY TIC A L BA TA Compound______________ C alculated_______ Found iv-5 € , 82.3s H, 9 .8 fi3 8 1 .5 , H, 9.7 IY-6 C, 8 1 .9s H, 10 .3 € , 8 0 .8 , H, 9 .7 IV-7 C, 8l.fcj H, 9 .6 € , , H, CHAPTER I I I T H E SYNTHESIS O F SEVERAL N E W T H Y R O X IN E ANALO G S. For reasons d iscu ssed in Chapter I I , the preparation o f thyroxine analogs o f stru ctu res III-L through 111-1$ was deemed d e s ir a b le • 1 . Simple Methyl B in itr o Analogs I I I - l ^ A'C > 1 C t k o ^ o / / A/C, I I I -2 o // ii i - i * o f , aa?2 H I - 5 // l" 3 A*>v 0 I I I -6 if /U?V c C(u /JO*, / ■ ^ I I l )0 Higher A lk yl S in itr o Analogs c# I I I -8 aA O C'lfr A J O ', cJJ v o „ _ // £ > < p \ U £ c//^ o~ o c J ^ c //3 Bxalkoxy B in itro Analogs i i i - i o /jp^ o m o ^ y * £ > * c/^ c x Mi JL/O^ 1 1 1 - 1 1 £% //* A)0 1 1 1 - 1 2 ' V l > 0 ^ 3 CdjCH^o 111-13 c ? // o c iU M ^ j jjo^ it. lo d in a ted Analogs i i i - i i ^ j /UP. 1 1 1 - 1 5 ocM i & a l3 x I 1 2 ! The gen eral method o f sy n th esis o f th ese d in itro d ip h en y l ; eth er analogs was th a t reported by the Glaxo workers (32) w ithout m o d ifica tio n , or in se v e r a l cases u sin g the author’s s lig h t m o d ifica - 1 t io n s , and in one c a s e , I I I - l , using the more e x ten siv e m o d ifica tio n s : o f Bruice (3 3 ). As i s e v id e n t, the "unprime” rin g was h eld co n sta n t, j w ith only the p h en olic ring being v a r ie d . U n fortu n ately, the y ie ld s . obtained in th e coupling rea ctio n in v o lv in g a lk y l su b stitu te d hydro- j quinones and phenols are found to be lower than coupling rea ctio n s in volvin g the u n su b stitu ted sp e c ie s (3 3 ). However, the y ie ld s may be in creased appreciably by using a sev er a l molar excess o f phenol as compared to d in itr o compound. In a few cases the author used an ex cess o f th e phen ol, thereby in crea sin g the percentage y ie ld , but j in most cases equimolar amounts o f the phenol and d in itr o compound t j were u sed . In most cases the la t t e r procedure was d ic ta ted by the i lim ite d q u a n titie s o f the various su b stitu te d phenols and hydro- ' quinones. By c o n tr a st, the dinitro? compound was r e a d ily a v a ila b le I through a new sy n th e tic procedure to be described s h o r tly . ! The Bruice m o d ifica tio n (33) o f the Glaxo workers m o d ifica tio n , o f the Ullmann and Nadai (50) sy n th e sis o f diphenyl eth ers has the advantages o f providing a reasonable y ie ld o f very high q u a lity product wherein ex cess pyrid in e and ph en olic substances are elim - l ! in a te d . These contaminants otherw ise in h ib it c r y s ta lliz a tio n o f the i d in itro d ip h en y l eth er product to such an e x ten t th a t column chromato- !3 I 1 I graphy o f the crude ta r i s n ecessary to remove them. The author, ! by u sin g equimolar q u a n titie s o f phenol and d in itr o compound r e a c t a n ts, and m odifying the procedure to elim in a te p y rid in e, achieved ! I a product o f a q u a lity approaching that o f Bruice*s method, but j w ithout recourse to the more lab oriou s methods o f th a t worker. At the same time the a lk y l-s u b stitu te d phenol i s conserved, although a t the expense o f o v e r a ll y ie ld . Two methods were used to prepare ii-hydroxyhydrocinnamic a cid : Method 1 . O verall y ie ld : 31$ c o o y — c / ^ c ° ° H c z / ^ c o o a / ^ / / o C /% c o o // Method 2 . O verall y ie ld : k0% C ~\ u o o S > I/S? ,/ y ' 7 ~ \ ^ ^ //Q CJ/^ c * C O O /1 Z / * I The f i r s t method was lim ite d to sm a ll-sc a le q u a n titie s , w hile the second was adaptable to very ra p id , la r g e -s c a le p rep aration s, in so fa r as the acetoxy in term ed iate, 111-21, was n ot is o la t e d . I 11-19 was d in itr a te d by the method o f Bruice and Kharasch to give 111-22, which was subsequently e s t e r if ie d to y ie ld H I -2 3 , fo llo w in g a m odified F isch er tech n iq u e. Ill /7~~\ *° t > / C/% 1 C #kc ' d y ' * X -J y ~ C /£ c //:z c & o tf ~~> / / o / A c / c \= z J ^ n v #c ^ A /a * V \ X x - 111-26 was prepared u sin g the s p e c if ic d ir ec tio n s o f Baker and Brown in th e ir m o d ifica tio n o f the Elbs p e rsu lfa te o x id a tio n on I p h en ols. The interm ediate a ry l s u lf a t e , 111-25, was n ot is o la te d ! | in pure formj ra th er, i t was m ethylated, follow ed by acid cleavage ! to give 111-26. ! I r ^ < ^ a s o ^ j ^ c h 3 c q ^ j i i I 111-26 was coupled w ith 111-23 to g iv e I I I - l u sin g the Bruice j m o d ifica tio n o f the Glaxo method. 1 cH* c! t ^ o : C ^ o jQ ^ o t) + E C -2 3 ' ------------- % ^ 0 / Q ^ O - ^ a i c ^ J ac A M J , 1 / v ^ ' : I I I -29 lik ew ise was prepared using the s p e c if ic d ir e c tio n s ; o f Baker and Brown. I l l -28 was is o la te d in crude form, m ethylated, 1 and then cleaved to give H I -2 9 . ; : „ J * , -) / 7 / 2 ' „ Ij s o ( j ) - o s o ~ a c [ J -------^ i n -2 9 w as th e n c o u p le d w it h I I I - 2 3 u s i n g t h e G laxo m eth od t o g iv e I I I - 2 . ^ 0 + J T T -J 3 — c //s o 0 ~ o -<f^ A)a^ - I l l -31 was obtained from the raanganeese d ioxide oxid ation o f and a c id ifie d s o lu tio n o f 2 ,3-d im eth ylan ilin iu m h yd roch lorid e. The r e s u ltin g quinone was steam d e s t ille d , and then reduced to the hydro- quinone, I I I - 3 2 . c # i c ^ , Cf*i < ~ ^ 3 y J + _ V t Z . ) = ( 2 !v * . 3 I I I -32 was coupled w ith I I 1-23 to give I I I - 3 , fo llo w in g the Glaxo method. S o ? 1 p \ ,? Aio 111-35 was prepared from I I I -33 by the Baker and Brown method, w ithout is o la tin g the interm ediate a r y l s u lf a t e . The s u lfa te was m ethylated and cleaved to give I I I -3 5 • 1 6 I This same substance was reported by Bruice who prepared i t by a more i j ela b o ra te method in v o lv in g the in term ed iates I I I -3 6 through 111-39 3 ! ach ievin g an o v e r a ll y ie ld o f $?> % • c t f i C .//3 7 7 7 --j 3 ------- > ■ — — * a ^ o ^ cM i ■ ~ s > -------— = > C -tfjO C /%°xJ^)0M J. c # 3 oJ?7~ C/% 111-35 was coupled w ith 111-23 to produce I I I -l;, fo llo w in g the | ! ' J i Glaxo method. \ I i , * > ^ T - o I 1 ( 3 0 / 0 } r ~ \ / / \ \ ,f ! p Z T :- ^ S “ + 2 Z T - J 3 -------*> a 4 0 y y (9~ ) ~ } c '/^ c /^ c ^ o Cx / y j ; c / ^ ✓ v ^ j s 1 I 3 ,5 -X y len o l was used to prepare I I I - l£ u sin g the Baker and ! ' i : Brown method. j C 'l y C A ~— ' b 6 ^-—{ o s * jA r*J ~ Qi 0,C y 0 ^ \ ^ ^ \ i ‘ : 1 ir c -1 ^ was coupled w ith 111-23 to giv e I I I - 5 , using the Glaxo method. \ ! c /^ a a . ! - ' f c s / z * j r r - n 2 3S - X y le n o l w as c o n v e r te d t o I I I - ljU b y t h e B ak er and Brown m e th o d . M e t h y la t io n , f o llo w e d b y a c id c le a v a g e g a v e I I I-itf> . ( r ii ^ C"j TLI-bS was coupled w ith I I I -23 to y ie ld I I I - 6 , u sin g the Glaxo method. c / i * °i- 772- - * s - / j 3 - < ^ - C ^ S OQ 4/Q > III-U 8 was prepared by reducing .l,ii-d in itr o d u r e n e , follow ed by d ia z o tiz a tio n and acid h y d r o ly sis. The o v e r a ll y ie ld was hh%. C /4 C /£ C % 'c^ III-1;8 was reacted w ith I I I -23 to g ive l'H -7 , a f te r the Glaxo method. c / 4 , c / 4 j z c - w + i t L ^ v i c / 4 Thym ol was o x i d iz e d b y t h e B ak er and Brown m eth od t o g iv e i I * I I I -50 which was m ethylated and cleaved to give I I I -5 l» c > 4 Ho m - 5 l was coupled w ith 111-23 to giv e I I I -8 by the Glaxo method. u NO*- J Z T - SV + 7 T T - 2 S fJLC -C ' C,4 t A \ — t C/AO</ N C /A C ' OC* A/c- ,LC t f i 111-52 was coupled with 111-23 to g ive I I I - 9 , a f te r the cfh, -{3-0^4 Glaxo procedure. ,<c ' f c r c 'i . ? ^ A ? —\ < 2 oJ% " + J. A)0, -c/-4 “ i " 1 * 6 ' % 2 ,3 - Dimethoxyhydroqtiinone, I I I -5 6 , was prepared by the method o f Baker and Savage, whereby p y r o g a llo l c a rb o x y llic a c id , I I 1-5 3 , was dem ethylated to give I I I -5 ii, then decarboxylated under vacuum to I I I -5 5 , follow ed by the Elbs p e rsu lfa te o x id a tio n and hy d r o ly sis to the hydroquinone. 6)ix oM ocfA pctij ^ / A o { ~ \ - c d ^ w ^ o ^ J ^ - c c H N y / — * / / o r y — » i <r» H o// '}✓ /•/ O c/A I I I - 5 6 w as c o u p le d w it h I 1 1 -2 3 t o g iv e I I I - 1 0 . j I c /jo ocM f | H r - S Z A -72Z-J3 - - - - - ^ J | j I i 2 ,5-Dimethoxyhydroquinone was prepared by the method o f | Inoevenagel and B uckel, rea ctin g methanol w ith l,h-benzoquinone in | the presence o f fu se d , anhydrous zin c ch lo rid e and the reducing the ! I S f in a l quinone product to the hydroquinone. octtj ° ^ Z==\=,r~, ~ --------- * \h O $ ~ \ - O H ocffi ocH i o c/j i I I I -58 was coupled w ith I I I -23 to I I I -1 1 . A)<2 o c % A )O v 0 JIT- S-<p y - 7 7 T . J 3 — * ^ o ^ ~ ^ o ~ v y ~ a / ^ c M c ~ - o c , , * fy P 2,5-Diethoxyhydroquinone was prepared by the method o f I ' " j Knoevenagel and Buckel, rea ctin g eth a n o l w ith 1 , 1 4 ,-benzoquinone in : the presence o f zin c c h lo r id e , follow ed by red u ction to the hydro- I j quinone* i O O ^ r Q C z. / • / r 2 0 111-60 was coupled w ith I I I -23 to g iv e H I-1 2 by the Glaxo procedure. 0 Cj4 ^ T T T - ( o 0 f T H r W K )O x O 2 ,5-Di-n-proxyhydroquinone was prepared according to the d ir e c tio n s o f Knoevenagel and B uckel, in a manner analogous to th a t used in the two preceding p rep a ra tio n s. cMSlk T i r - 5 7 — I— \-o - a & ( o j n z - z z ( J ! h cljcllp I I I -62 was coupled w ith I II -2 3 to g iv e I II -1 3 using the Glaxo procedure. o d U A & s jjo ^ p l n r - 6 2 , - f - m r - z s — ^ O l p l ^ O jJ Q I H - 1 was reduced, te tr a z o tiz e d , iod in ated and cleaved to ‘ 1 i g iv e I l l - l i t , w ithout is o la tin g 111-61* or I I I -6 5 . I cU* uiL P j J Z E - / ___ ^ j w k * 7 ” -J- p ~ x 2 1 I I I -11 was reduced, te tr a z o tiz e d , io d in a te d , and cleaved to g iv e I I I - 1 5 , w ithout is o la t in g I I I -66 or 111-6?. O t\) 3 V jk ' E C - 1 1 a / / ocff fjtf o ctlj X o cjJx C > cJj^cds X 2 2 R E A G E N T S A N D S O L V E N T S ^ t ! 1 . U nless otherw ise s p e c if ie d , a l l re a g e n ts, so lv e n ts and ; s ta r tin g m a teria ls were o f or ’’Reagent” grade, or were p u r ifie d by standard m ethods. 2 . ii-Hydroxyben z aldehyde was purchased from Matheson, Coleman and B e ll. | 3 . 2 ,3-D im ethylanilinium hydrochloride was generously furnished by Mr. Franklin B illig . o f American Potash and Chemical Co. \ h» 3 , 5-X ylenol was purchased from Eastman Kodak Co. 5 . 2 ,6-X ylen ol was purchased from Eastman Kodak Co. , 6 . 2,f>-Xylenol was prepared by su b jectin g p -xylen e to su lf© n a tio n , follow ed by mixed potassium hydroxide-sodium hydroxide fu s io n . A c id ific a tio n , follow ed by steam d i s t i l l a t i o n gave a product ; o f e x c e lle n t q u a lity . This method was adapted from the g en era l r ea c tio n as given by F ie se r (1 $ ). j 7• D in itrod u ren e, l,U -d in itr o - 2 ,3 ,5 ,6-tetram eth ylbenzen e, was i I obtained as a crude stu d en t p rep aration , and was r e c r y s ta lliz e d j from methanol a fte r tr e a tin g the h ot so lu tio n w ith d eco lo rized ch a rco a l, follow ed by f i l t r a t i o n . j 8 . Thymol, 2-isop rop y1 -5 -m ethylphenol, was purchased from Baker and Adamson. ■9. 2 ,6 -D i-t-b u ty lp h e n o l was generously furn ish ed by E thyl C orporation. 23 2 ,6 “M -i«-propylphenol was generously fu rn ish ed by E thyl Corporation* 2,5-M -t-b u tylh yd roq u in on e was purchased from Eastman Kodak Co. y E X P E R IM E N T A L , li-H Y D R O X Y H T D R Q C IN N A M IG A C ID Method 1 . : li-Hydroxyhydrocinnamic a c id , 111-19, was prepared according to the procedure o f F . Konek and E . Pacsu (3k)• Method 2♦; Forty grams o f f in e ly powdered, anhydrous U-hydroxyben- zaldehyde were mixed w ith 6 J U g • o f fr e s h ly fu sed and f in e ly ground sodium a c e ta te . This m ixture was s tir r e d in to 100 m l. o f a c e tic anhydride in a 1 L. round-bottom f la s k . The fla s k was clo se d w ith an a ir condenser, 30-ii0 cm. in le n g th , atop which was placed a drying tube containing anhydrous calcium c h lo r id e . The m ixture was allow ed to r e flu x g en tly a t lijO-5Q0 fo r 2k hours. The crude product c o n sistin g o f U-acetoxycinnam ic a c id , 111-21, was d i f f i c u l t to h an d le, forming a hard, amorphous g la ssy substance on co o lin g . A ccordingly, i t was poured slo w ly , w h ile s t i l l warm, in to 1 L. o f water in a 2 L. beaker w ith rapid m echanical s t ir r in g . At the same time th a t the product was being added to the w ater, 150 g . o f sodium hydroxide p e lle t s were added s lo w ly . The temperature o f the m ixture r o s e , but a fte r i t had subsided to 6 0 °, and w ith continued rapid s t ir r in g , 60 g . o f Raney n ic k e l a llo y was added in sm all p ortion s over a four hour p erio d . The temperature was m aintained between 60 and 70° by 1 / The analyses reported here were performed by Mr. W.J. Schenck o f t h is la b o ra to ry . The a n a ly tic a l data are recorded a t the end o f th is s e c tio n . M elting p o in ts are u n corrected . I warming the m ixture on the steam b ath . A fter a d d itio n o f the Raney I | n ic k e l a llo y was com pleted, the m ixture was warned to 90-95° fo r l ■ two hours w ith continued s tir r in g and p e rio d ic a d d itio n o f d i s t i l l e d j w ater to compensate fo r evaporation l o s s e s . F ive grams o f d e c o l- | o rizin g charcoal were added, and the m ixture s tir r e d f iv e m inutes j lo n g e r , and then f ilt e r e d through a sin te r e d g la ss fu n n e l. The black i I resid u e was washed w ith two 50 m l. p o rtio n s o f h ot tw o-percent sodium I J hydroxide s o lu tio n . The f i l t r a t e and washings were combined and then I j a c id ifie d to Congo red w ith conc. h yd roch loric a c id , c h ille d , and the product ex tra cted w ith e th e r . A fter evaporation o f the e th e r , the ; brown resid u e was d i s t i l l e d under vacuum to g iv e a w hite s o lid . The ' s o lid d i s t i l l a t e was d isso lv e d in benzene w ith warming, g iv in g tr a n s- ; parent w hite n eed les upon the ad d itio n o f c ir c a o n e-fou rth volume o f " S k elly so lv e Btt. The o v e r a ll y ie ld was 22 g . , 1*0.1$. The preparation o f U-acetoxycinnam ic acid i s th a t as given by F.von Konek and E.Pacsu (3I 4) . The red u ction u sin g Raney n ic k e l a llo y in base i s adapted from the procedure reported by Schwenk and Papa fo r the red u ction o f sim ila r substances ( 3 5 ). The lite r a tu r e y ie ld fo r the P erkin-type condensation i s 50$, w h ile the b a sic Raney n ic k e l J alloy red u ction o f sim ila r compounds proceeds in c ir c a 80$ y ie ld . ! ;The m eltin g p o in t reported fo r ii-hydroxyhydrocinnamic a cid i s 128-30° | (3 6 ) , found 126- 296 . ! ETH Y L 3,5-DINITRO-k-KYDROXYH Y D R O C IN N A M A T E | ! 3 >5~Dinitro-li-hydr6xyhydrocinnamic A cid, 111-22. This i j substance was prepared fo llo w in g the method reported by Bruice (37) using a c e ty l n it r a t e , generated a t low tem perature, as the a c tiv e ' n itr a tin g agent* Observed m eltin g p o in t 1 3 6 °, L it . 137*5° (3 8 ). | Y ield ob tain ed , 73 $ , l i t . lh% (3 7 ). i : E thyl 3 ,5 ‘-dinitro~J4-hydroxyhydrocinnamate. III-2 3 « Into a | 100 m l. f la s k were placed 5.12 g . (0 .0 2 m o l.) o f 111-22 and 1*0 m l. i o f ab s. e th a n o l. Three m l. o f conc. s u lfu r ic a cid were added drop- w ise w ith sw irlin g o f the f la s k . A Vigreux column w ith calcium ch lo rid e drying tube was attached to the f la s k , and the m ixture ; reflu x ed on the steam bath fo r fou r h ou rs. The h ot so lu tio n was | f ilt e r e d and d ilu te d w ith ten m l. o f w ater, allow ed to c o o l, and t • r e fr ig e r a te d . Golden n eed les o f 111-23 appeared and were f ilt e r e d ! o f f . S u ccessiv e d ilu tio n s w ith w ater, fo llo w ed by a f in a l eth er e x - | tr a c tio n o f the mother liq u o r and evaporation o f the eth er gave a ; crude y ie ld o f 100$. The a ir -d r ie d crude product was d isso lv e d in a ! |minimum volume o f chloroform , and two or three volumes o f " S k elly - i jso lv e C” were added w ith wanning on the steam b ath . O n c o o lin g , p ale |y e llo w p la te s o f 111-23 were d ep o sited , m eltin g p oin t 6 7 -6 9 °, l i t . i |7 ii-5 ° . The y ie ld i s g rea ter than 95$, L it . 69$ (3 7 ). T ^ is method, i I using s u lfu r ic a c id , i s an adaptation o f th a t reported by F isch er I •and S p eier (3 9 ). 2 7 ETH Y L 3 , 5-DINITRO-l*- (3 « -M ETH YL-1*' -M E T H O X Y F H E N O X Y ) - \ H Y D R O C IN N A M A T E \ ! ! 3-Methyl~l*-me thoxyphenol. I I 1 -2 6 . This interm ediate was j prepared from o-rcresol, 111- 21*, using the s p e c if ic d ir e c tio n s given i by Baker and Brown (1*0) fo r th e ir m o d ifica tio n o f the Elbs p e rsu lfa te | o x id a tio n on p h en ols. The interm ediate s u lf a t e , 111-25* was extracted 1 I in to 95$ e th a n o l, a fte r air-stream evaporation o f the a c id ifie d and i : | j eth er ex tra cted crude o x id a tio n m ixture. The d ir e c tio n s o f th ese i | . | authors are not s p e c if ic ,as to the ex a ct procedure fo r sep aratin g •i _ • ' 5 the interm ediate a r y l s u lf a t e . This interm ediate was then m ethylated ; and cleaved w ith acid to the methoxy phenol in accordance w ith th ese ! author’s s p e c if ic d ir e c tio n s . The product, a fte r r e c r y s ta lliz a tio n \ from w ater, m elted a t 1*1-3°, L it . 1*6-1*6.5 ° (1*0). Subsequent re cr y s t a lliz a t io n from b en zen e-"S k ellysolve €'• f a ile d to in c r e a se ,th e m elt^ I ing p o in t. Y ield 20$, L it . 23$ (1*0). 1 E thyl 3 ,5 -d in itr o -l* -(3 1 -methyl-1*1 -m ethoxyphenoxy)-hydrocin- I namate. I I I - l . This substance was prepared from 111-26 and 111-23 | in 33$ y ie ld , u sin g a one molar ex cess o f the p h en ol. The method f o l - : lowed was th a t o f Bruice (33) fo r the preparation o f a sim ila r com- ipound. Three re c r y s ta lliz a t io n s from chloroform -!lS k e lly so lv e C " gave ;a product m eltin g a t 7 5 -7 6 °. A s a tis fa c to r y a n a ly tic a l sample was ob- | ta in ed only a fte r chromatography on a 15 cm. column o f f in e alum ina I ju sin g m ethanol/benzenej 1/ 5 , follow ed by evaporation and one a d d itio n a l 28 r e c r y s ta lliz a t io n from ch loroform /" S k ellysolve € nj 1/5* ETH YL 3 *5-DINITRO-li-( 2' -METHYL-ii' -M E T H O X Y F H E N O X Y ) - HYDROCINNAMATE 2-Methyl-k-metho:xy p h en ol. I I 1-29« This interm ediate was pre pared from m -c re so l, 111-27* u sin g the s p e c if ic d ir e c tio n s o f Baker and Brown (I4 .O ) in th e ir m o d ifica tio n o f the Elbs p e r su lfa te o x id a tio n . The d e t a ils o f the procedure fo llo w ed are sim ila r to those described fo r the preparation o f I I 1-2 6 . A fter two r e c r y s ta lliz a tio n s from water* the product m elted a t 7 0 ° , L it . 71° (U o). The y ie ld obtained was 17#* L it . 33$ (J*0). E th yl 3 * 5 -d in itr o -ii-(2 1 -m eth yl-li1 -methoxyphenoxy ) -hydro- cinnam ate. I I 1 -2 . This substance was prepared from 111-29 and 111-23 in hP% y ie ld , fo llo w in g the gen eral d ir e c tio n s o f C layton, Green and Hems (3 2 ). Two minor m o d ifica tio n s o f th e ir method were introduced and which afforded a more re a d ily c r y s ta lliz a b le product. The f i r s t m o d ifica tio n co n sisted o f adding f if t e e n m l. o f conc. h ydrochloric acid to the crude coupled product in eth an ol-w ater- pyrid in e s o lu tio n p rio r to e x tr a c tin g the product w ith chloroform . Presumably t h is h eld the p yrid in e in the aqueous phase, thereby preventing contam ination o f the coupled product in the chloroform phase. The second m o d ifica tio n in volved evaporating the chloroform e x tr a c t to dryness and h eatin g to 80° fo r three hours a t 0 .5 mm. Hg p ressu re. The r e s u ltin g brown o i l was rep eated ly ex tra cted w ith 2$ m l. p ortion s o f " S k elly so lv e € " , to which 1-2 m l. o f chloroform ■ A had been added. This e x tr a c tio n procedure was continued u n t il only a b la ck , tarry resid u e rem ained. Further r e c r y s ta lliz a tio n s were from f iv e percent chloroform in " S k ellysolve € " . Repeated recry s t a lliz a t io n s from t h is so lv e n t p a ir , w hile y ie ld in g a constant m elt ing product o f good c o lo r , f a ile d to y ie ld samples a ffo rd in g the co rrect elem en tal a n a ly s is . €dnsequently the substance was chroma tographed on a 15 cm. column o f powdered alumina using m ethanol/ benzene; 3 /1 > w ith e lu tio n by a b s. m ethanol. This elu en t,w as evap orated in a covered d ish and r e c r y s ta lliz e d from ’’S k e lly so lv e € ,f. The m eltin g p o in t o f the pure sample was 98-9°• ETHT1 3,5-BINITRO-li-(2* ,3 ’ -DIMETHTL-h! -H YDROXYFHENOXY)- H Y D R O O IM A M A TE 2 ,3 -Dimethy1 - 1 ,U-benzohydroquinone. I l l -3 2 . This compound was prepared by the gen eral method o f Clark ( h i) , as given by F ie se r l (h2) fo r t h is gen eral c la s s o f su b stan ces. A h .0 % y ie ld o f the in te r m ediate quinone formed by the manganese d ioxid e o x id a tio n o f 2 , 3 -d i me thy la n ilin iu m hydrochloride was obtained only a f te r s e v e r a l m l. o f 10$ potassium dichrornate so lu tio n were added to the m ixture during steam d i s t i l l a t i o n . This in d ica ted th a t the d ir e c tio n s , i f follow ed as such, would g iv e an incom plete o x id a tio n . The quinone product obtained was s t a b le , m eltin g a t 5 1 -2 °. M t . 55® (i+3). Two g . (O.Olij? m o l.) o f the quinone were d isso lv ed in g la c ia l a c e tic acid 3 0 and reduced with hydrogen a t atm ospheric p ressure over 1 0 % p a lla d ized ch a rco a l. The reduced product was f il t e r e d , a sp irated to dryn ess, then chromatographed on a 15 cm. column o f f in e alumina using meth anol/benzene; 1 /5 . The c o lo r le s s e fflu e n t was evaporated in the dark in a covered d ish . A y ie ld o f 2$% was ob tain ed . M elting p oin t 2 16-20°. L it . 221° a fte r re c r y s ta lliz a t io n from w ater (1*3) • E th yl 3 , 5 -d in itr o -i4 -(2 1, 3 1 -d im e th y l-!’ -hydroxyphenoxy)~ t hydrocinnamate . I l l - 3 . This substance was prepared in J > 0 % y ie ld from 111-32 and 111-23 u sin g the gen eral method o f -Clayton, Green and Hems (3 2 ), w ith the excep tion th a t a rea cta n t r a tio o f 1 /1 was u sed . The crude product obtained from d e s sic a tio n o f the chloroform e x tr a c t o f the rea ctio n m ixture was ex tra cted w ith chloroform / ’•S k elly so lv e iC ” ; 1 /1 by volume, and the crude product r e c r y s ta lliz e d from the same so lv e n t p a ir . The chromatographie preparation o f an a n a ly tic a l sample used the same procedure as described fo r IT I-2 . M elting p o in t, 202°. ETHIL 3 ,5-DINITRO-U-(3* ,5 ’ -DIMETHYL-!' -M E T H O X Y FH E N O X Y ) - H Y D R O C IN N A M A T E 3 ,5~Pim ethyl-Jj-m ethoxyphenol. 111-35. This interm ediate was obtained in 20$ y ie ld from .2 ,6 -x y le n o l, 11 1 -3 3 , fo llo w in g the s p e c if ic d ir e c tio n s fo r the preparation o f t h is substance as given by Baker and Brown (1*0) in th e ir m o d ifica tio n o f the Elbs p e rsu lfa te o x id a tio n . The crude product was r e c r y s ta lliz e d from equal p arts o f 31 b en zen e-" S k ellysolve €" and m elted a t 7 9 -8 1 °. l i t . 83° (!*0), 8it-5*° (3 3 ). No l i t . y ie ld was reported fo r th is method o f prepara tio n o f 111-35- E th yl 3 j5 -d in itr o -U -(3 * , 5 ' -d im eth y l-1 1 -methoxyphenoxy) - hydrocinnam ate. 1 1 1 -1 . This compound was prepared by the method o f C layton, Green and Hems (32) using a 3»li molar r a tio o f 111-35 to I I I -23 to g iv e a 60 % y i e l d . The d essic a te d and evaporated chloroform e x tr a c t o f the crude m a teria l was ex h a u stiv ely ex tra cted w ith b o ilin g " S k ellysolve € " . The product was r e e r y s ta lliz e d from t h is s o lv e n t. M elting p o in t was 1 1 3 °. A n a ly tica l samples were prepared by the same procedure as described fo r I I I - 2 . , ETH Y L 3 95-DINITR0-U- (2' ,6 » -DIMETHYL-!*' -M E T H O X Y FH E N O X Y ) - 1 H Y D R O C IN N A M A T E 2 s6-Dim ethyl-ii-m ethoxyphenol. I II -ij2 . This interm ed iate was obtained in 21$ y ie ld fo llo w in g the s p e c if ic d ir e c tio n s o f Baker and Brown (ItO) in th e ir m o d ifica tio n o f the Elbs p e r su lfa te o x id a tio n . The crude product m elted a t 73°5 r e c r y s ta lliz a tio n from b o ilin g " S k elly so lv e C " afforded a product m eltin g a t 7 5 -6 ° , l i t . 77 (!+0). No y ie ld was rep orted . E thyl 3 s5 -d in itr o -i* -(2 1, 6 1 -dim ethyl-!*1 -methoxyphenoxy )- HydroeiMhMaid. I l l - 5 . This substance was prepared from III-i*2 and I I I -23 in h0% y ie ld u sin g the gen eral method of C layton , Green and Hems (3 2 ). A rea cta n t r a tio o f 1 .5 /1 fo r III-!* 2 /III-2 3 was u sed . 32 The crude product was ex tra cted w ith h ot ’'S k e lly so lv e C” containing a tra ce o f chloroform . Subsequent r e c r y s ta lliz a tio n was from t h is i so lv e n t p a ir . A n a ly tic a l samples were prepared by the same procedure as described fo r I I I - 2 . M elting p o in t 206°. i ! ETHYL 3 > 5 -D IN IT R O -li-( 2 1 , 5 ' -B E fE TH Y L -V -METHOXYPHENOXY)- I H Y D R O C IN N A M A T E I 2 , 5-Dim ethyl-lt-m ethoxyphenol. III-1 |5 » This interm ediate was prepared from 2 ,5 -x y le n o l, I I I -I 43, in 20% y ie ld by the general method o f Baker and Brown (l+O). These authors did n ot rep ort th is ' s p e c if ic su b stan ce. The product was r e e r y s ta lliz e d from ”3 k e lly - 1 so lv e C ” con tain in g a tra ce o f chloroform . M elting p o in t found, I 8 5 -7 °, L it . 90° (1+5). i i !•- j E thyl 3 ,5~-dinitro-l+-(2 * ,5 ' -dimethyl-i+* -methoxyphenoxy ) - I hydrocinnamate. I I I - 6 . This compound was obtained in 2k% y ie ld from III-i+5 and 111-23 by mean o f the gen eral method o f S la y to n , 1 j Green and Hems (3 2 ), using a rea cta n t r a tio o f 1 / i . The crude product | was ex h a u stiv ely ex tra cted w ith b o ilin g ’’S k e lly so lv e C " and subse- | quently r e e r y s ta lliz e d from t h is so lv e n t w ith a tr a ce o f chloroform i added dropwise as needed to prevent " o ilin g out" o f the product. ■ ' | A n a ly tica l samples were prepared by the same procedure as described 1 fo r III -2 * M elting p o in t 106°. | ETH Y L 3 i 5-DIllITRO-l|-(2l ,3 ’ , 5* ,6 ‘ -TETRAM ETHYL-M - HT- | B R O X Y P H E N O X Y ) -H Y D R O C IN N A M A T E j i 2 ,3 >5 j6-T etram eth yl-l ,l*-benzohydroquinone. 111-1*8. Ten g . o f ; ; III-li.6 were d isso lv e d in 5© m l. o f a b s. methanol w ith warming on the i ' i : i steam b ath . The so lu tio n was f il t e r e d in to a 200 m l. f la s k , and a i t - i i | sm all s p a tu la fu l of Raney n ic k e l c a ta ly s t was added. A 35 cm. Vigreux j | column was attached to the f la s k , and f iv e m l. o f 85$ hydrazine hydrate! \ | i was allow ed to drip down the column very slo w ly from a sm all dropping j J i fu n n e l. The ad d ition took approxim ately ten m in u tes, and was accom- ; panied by an extrem ely vigorou s r e a c tio n . A fter the re a c tio n had ; su b sid ed , a drying tube con tain in g calcium ch lo rid e was a ffix e d to r ! the top o f the column, and the m ixture was allow ed to r e flu x overn igh t I | on the steam b ath . The r e s u ltin g c o lo r le s s so lu tio n was f ilt e r e d j I w hile s t i l l h o t, and on co o lin g d ep osited c o lo r le s s n eed les o f 111-1*7, j ; i ! which ra p id ly darkened on sta n d in g . A p o rtio n o f the crude product ; was d ried under vacuum, and a sample weighing 2 g . (0.0122 m o l.) was ; i ^ i : d isso lv e d in 20 m l. o f 10$ s u lf u r ic a c id . The acid s o lu tio n was I i ’ j | 1 I cooled to 0° in an ic e bath and w ith m echanical s tir r in g 1 .7 5 g . o f j I \ ; j | sodium n i t r i t e was added over a 15 minute p erio d . The s o lu tio n was j * ! ! placed in a r e fr ig e r a to r a t -5 ° fo r 90 m inutes fo llo w in g which i t was I i * ; poured in to 150 m l. o f w ater. The aqueous so lu tio n was heated to j i ' | b o ilin g fo r f iv e m inutes, cooled in an ic e b ath , ex tra cted w ith e th e r , I and the e x tr a c t dried over anh. magnesium s u lf a t e . The d ried ex tr a c t 'was f il t e r e d and evaporated in an open d is h . The y ie ld o f w h ite , • 3k c r y s ta llin e product was 0 .8 9 g . (1 4 $ ). M elting p o in t 2 00-10°, l i t . ! 210-2J a (1*6). i . E th yl 3 jS -d in itr o -k -te 1 ,3 ' ,£* ^ '-te tr a m e th y l-i* '- hydroxy- j - - - - - - - - - - - - - — - - - - - - - - - ;- - - - - - - - - - - - - - - - - - - - * - - - - - - - - - - - - - - - - - - - - - - - J phenoxy) -hydrocinnam ate. I I I - 7 . This substance was prepared in 28$ | y ie ld from I I I -1*8 and 111-23 by the gen eral procedure o f S la y to n , I Green and Hems (3 2 ). The crude chloroform exrtract was trea ted in a i fa sh io n sim ila r to th a t described fo r I I I - 2 . A 1 /1 rea cta n t r a tio was u sed . A n a ly tic a l samples lik e w ise were prepared as described under I I I - 2 . M elting p o in t 61|°. ' E TH Y L 3 ,5-BINITRO-J*-(2' -METHYL-5’ -ISOPROPYL-it' -M ETH O XY- | F H E M O X Y )-H Y B R O G IN W A M A T E I | 2-Me thy 1 - i*-m ethoxy-5-i sopropylphenol. I I I - 5 1 . This in te r - i I m ediate was prepared by the gen eral method o f Baker and Brown (1 0 ). i j The product was o ily and f a ile d to c r y s t a lliz e from common laboratory i i s o lv e n ts . However, the product c r y s ta lliz e d from a r e fr ig e r a te d i i J aqueous s o lu tio n a fte r the hot so lu tio n tw ice had been tr ea ted w ith j o n e-h a lf gram o f d eco lo rizin g ch arcoal, follo w ed by f i l t r a t i o n . I I Even t h is product appeared crude and had a brownish co lo r j hence, i I the m a teria l was chromatographed on a 15 cm. column o f fin e alumina i ! using m ethanol/benzenej 1 /3 , w ith e lu tio n by the same so lv e n t p a ir . i ■ ! } i The product remained an o i l and r e s is te d c r y s t a lliz a t io n . i j ! E thyl 3 ,5 -d in itr o -ii-(2 '^m ethyl-51 -iso p ro p y l-it1 -methoxy- | phenoxy) -hydrocinnam ate. I I I - 8 . This substance was prepared by the 35 gen eral method o f C layton, Green and Hems (32) in 21% y ie ld , using j I I I -51 and 111-23 in 1 /1 molar r a t io . The product was worked up in i the manner described fo r I I I - 2 . Both e x tr a c tio n and r e c r y s t a lliz a - j ; tio n were made from b o ilin g " S k ellysolve C". A n a ly tic a l samples were j prepared by the method described fo r I I I - 2 . M elting p o in t 99°• \ ETH YL 3 ,5-MNITRO-1j-(2' , 5 ' -DI-t-BUTYL-V -H YDROXYPHENOXY)- i H Y D R O C IN N A M A T E j I ! \ E thyl 3 ,5 -d ln itr o « k -(2 1 ,5 ' -d i-t-b u ty l-i* ' -hydroxyphenoxy) - i " * ■ 1 - "» " ...I.-, ii.— — ■ ..... ...i n .i ,■ ■ j hydrocinnamate. I I I - 9 . This compound was prepared by the gen eral : method o f C layton , Green and Hems (32) in \H% y i e l d , u sin g 111-52 ; and H I-2 3 in k /1 molar r a t io . The product was worked up in the s : manner described fo r I I I - 2 . Both e x tr a c tio n and r e c r y s ta lliz a t io n j i _ ! I were made from b o ilin g ’’S k e lly so lv e C ” . A n a ly tic a l samples were i : J i prepared by the method described fo r I I I - 2 . M elting p o in t 12li-5°C. j : ETH Y L 3 , 5-DINITRO-lt-( 2 ' ,3-BIMETH0XY-li«-HYDROXYPHENOXY)- ' H Y D R O C IN N A M A T E < ; 3-Hydroxy-It,5-d±methoxybenzoic A cid. III-5 U . This in te r m ediate was prepared from g a l l i c a c id , I I I -5 3 , fo llo w in g the s p e c if ic j • d ir e c tio n s o f Mauthner (5 3 )• The reported y ie ld was 75$; the observed : I ■ ! ; y i e l d , $k%. I ; i I : 2 ,3-Dim ethoxyphenol. I l l -55» This phenol was prepared from ! ■ I I 1-61 fo llo w in g the d ir e c tio n s o f Baker and Smith (51t), who reported j • a y ie ld o f 83$ . The observed y ie ld was 65$. The observed b o ilin g : range o f 230-^0° agreed w ith th a t reported by Herzig and P oliak ( 5 5 ). • 2 , 3-Dimethoxyh.ydroquinone. 111-56. The s p e c if ic method o f ; Baker and Savage (52) was used in preparing t h is interm ediate from ; I I I -55 by means o f the Elbs p e r su lfa te o x id a tio n . From f iv e g . o f ; I I I -5 5 , th ese workers recovered 1-2 g . o f a dark, v isco u s o i l which i 1 they were unable to c r y s t a lliz e . The observed y ie ld was c ir c a 1-2 g . E th yl 3 , 5 -d in itr o -i4 - ( 2 1 , 3 1 -dim ethoxy-i;1 -hydroxyphenoxy) - ! hydrocinnamate. I l l -1 9 . This compound was prepared by means o f the ! g en era l method o f S la y to n , Green and Hems (3 2 ), m odified as in I I I - 2 . A 20% y ie ld was obtained by u sin g a rea cta n t r a tio o f l / l j 1 1 1 -5 6 / . H I - 2 3 • The product was chromatographed fo llo w in g the d ir e c tio n s in i I I I - 2 . R e c r y s ta lliz a tio n was from " S k elly so lv e €" con tain in g a sm all 1 amount o f chloroform . M elting p o in t, i* 8° . i j ETH YL 3 ,5-DINITRQ-U- (2« ,5 ' -BIMETHOXY-li* -H YDROXYPHENOXY)- J \ H Y D R O C IN N A M A T E 2 , 5-Dimethoxyhydroquinone. I l l -5 8 . This interm ediate was ; prepared from quinone and m ethanol in the presence o f zin c c h lo r id e , i according to the s p e c if ic d ir e c tio n s o f Knoevenagel and Buckel (ii7 ). ; The immediate product formed by the condensation i s the quinone, ! I I I -5 9 , which i s q u a n tita tiv e ly reduced w ith stannous ch lo rid e and ; hydrochloric a cid to g ive 111-65. Observed m elting p o in t, 161;°5 : l i t . 166° (li7 ). Observed y ie ld , 2$%', l i t . 32$. E th yl 3 ,5 -d in itr o - li-( 2 1 ,5 * -dime thojxy-ii’ -hydroxyphenoxy) - hydrocinnamate. I I I - 1 2 . This substance was prepared from I I I -65 and 111-25 by means o f the gen eral method o f C layton, Green and Hems (3 2 ). A 36$ y ie ld was obtained by using a rea cta n t r a tio o f 2 .1 9 /1 ; 1 1 1 -6 5 / I I I -2 5 . A n a ly tic a l samples were prepared by the procedure described fo r. I l l - 2 . R e c r y s ta lliz a tio n was from chloroform /',S k e lly so lv e C ” j 1 /1 , or from m ethanol/benzene; l / l . M elting p o in t, 205°• ETH YL 3 , 5-BINITR0-Jt- (2 ‘ ,5 ' -BIETHQXI-ii-HYDROXYPHENOXY )- H Y D RO CIM AM ATE 2 ,5-B iethoxyhydroquinone. I I I - 6 0 . This in term ed iate was prepared by stannous ch lo rid e-h y d ro ch lo ric a cid red u ction o f 111-61 formed from quinone and methanol in the presence o f zin c c h lo r id e , fo llo w in g the s p e c if ic d ir e c tio n s o f Knoevenagel and Buckel (1*7). The observed m eltin g p o in t was 13li-6®, l i t . 138° (1*7) • Observed y ie ld 21$, l i t . 3 0 % -(1*7). E thyl 3 , 5 -d in itr o -lj-( 2 ’ ,5* -diethoixy-l*1 -hydroxyphenoxy ) - hydrocinnamate. I l l -1 2 . This compound was prepared by the method o f C layton, Green and Hems (3 2 ). A 29% y ie ld was obtained by using a rea cta n t r a tio o f 2 /1 j 111- 60/ I I I -23* A n a ly tic a l samples were prepared by the procedure described fo r I I I - 2 . R e c r y s ta lliz a tio n was from chloroform /1 ’S k e lly so lv e C M ; 1 /5 . M elting p o in t 206-8° (d e c .) 38 ETH Y L 3 , 5-DINITRO-li- ( 2» , 5 • -BI-n-PROPOXY-lt’ -HYDROXY- i i P H E N O X Y ) -H Y D R O C IN N A M A T E J 2 ,5-Di-ri-propoxyhydroquinone« I I 1 -6 2 . This in term ed iate was ; prepared by stannous ch lo rid e-h y d ro ch lo ric acid red u ction o f I I I -63 i | formed from quinone and n-propanol in the presence o f anhydrous zin c l c h lo r id e , fo llo w in g the s p e c if ic d ir e c tio n s o f Knoevenagel and Buckel ( I 4 .7 ) . The observed m eltin g p o in t was 91-3°> l i t . 95° (-it? ) - Observed y ie ld was 16$ , l i t . 23$ (1 * 7 ) . ! E thyl 3 , 5-d in itr o -i* - (2 1, 5 1 -d i-n -propo3 ty-l*8 -hydroxypheno:xy ) - • hydrocinnamate. 11 1 -1 3 . This substance was prepared from 111-62 and H I -2 3 by the method reported by C layton , Green arid Hems (3 2 ). j A 26$ y ie ld was obtained by u sin g a rea cta n t r a tio o f 2 /1 j 1 1 1 -6 2 / H I - 2 3 . A n a ly tica l samples were prepared by the procedure described fo r I I I - 2 . R e c r y s ta lliz a tio n was from chlorofoiTO -"Skellysolve GM . I M elting p o in t 1 8 7 -8 °. j 3 ,E>-BIT0B0-l*-(31 -M ETH YL-1*5 -H Y D R O X Y FH EN O X Y )-HYDROCINN- ) | A M IC ACID < 1 | 3 , 5-D iiod o-l*-(3 * -m eth yl-1 * 1 -hydroxyphenoxy) -hydrocinnamic A cid. I I I -ll* . A 359.6 mg. sample (0.889 mmol.) o f I I I - l was d is - ; so lv ed in 20 m l. o f g la c ia l a c e tic a cid and reduced w ith hydrogen [ j over 10$ p a lla d ized charcoal a t room temperature and atm ospheric I p ressu re. The r e s u ltin g diam ine, 111-61*, was te tr a z o tiz e d , io d in a te d , 39 and cleaved to g iv e I II -I I 4, fo llo w in g the gen eral method o f B ru iee, Kharasch and W inzler ( 33)* The crude product was is o la te d from the g la c ia l a c e tic a cid -h y d rio d ic acid eleaveage m ixture and r e c r y s ta l liz e d s e v e r a l tim es from acetone/w ater; 1/ 1 0 . The y ie ld was 37$, m elting p o in t 196- 9° . 3 , 9-DIIODQ -i*-(2 1 ,5* -DIMETHQXI-ii.1 -H Y D R Q X Y FH EN O X Y ) - H Y B R O G IK N A M IG AG ID 3 , 9-D iiod o-it- (2 1 , 5 1 -dime thoxy-U1 -hydroxyphenoxy ) -hydro- cinnamic A cid, in - 1 5 . A ijO O mg. sample (0.93 mmol.) o f I I I - l l was d isso lv e d in 20 m l. o f g la c ia l a c e tic a cid and reduced w ith hydrogen over 10% p a lla d ized charcoal a t room temperature and atmos p heric p ressu re. Follow ing the gen eral procedure o f B ru ice, Kharasch and W inzler (33), the diamine was t e tr a z o tiz e d , io d in a ted and cleaved to g iv e I I I -1 9 . The d ried , crude product was chromatographed in acetone on a column o f fin e alum ina, thereby removing colored impu r i t i e s . The product was r e e r y s ta lliz e d from ace ton e/w ater j 1 /1 0 . M elting p o in t, IJ4 I-U 0 • Y ield 18$. TABEE I ANALYTICAL DATA Compound C a l c u l a t e d F ound ________________C H N __________ C H N I I I - l 56.48 4.95 6 .9 3 56.71* 5 .0 3 6.7 2 I I I -2 56.48 it. 95 6 .9 3 57i.l*5 5 .0 6 I I I -3 56.48 4 .9 5 6.9 3 56.51 4.67 n i - 4 57.46 5 .2 6 6 .7 0 58.75 5.1*3 n i - 5 57.46 5 .2 6 6 .7 0 57.21* 5.62 I I 1-6 57 .46 5 .2 6 6.70 57.29 5.1*1* I I 1-7 58.38 5 .5 6 6.1*8 111-51 73.57 8.8 9 0 .0 0 I I I -8 58.38 5*56 6.1*8 56.1*2 5 .1 1 6 .4 8 I I I -9 . 61.48 6 .5 6 5.7it I I I — 10 52.34 it.59 6.1*3 I I I - l l 5 2 .3 4 4.5 9 6.1*3 52.1*9 4 .8 2 I I 1-12 51*.31 5.17 6 .0 3 111-13 56.10 5 .6 9 5 .6 9 I l l - l i t I , l*8.itl I , I I I -1 5 I , 44.56 I , ' C H A PT E R IV j I T H E FERRIC CHIORIBE OXIDATION O F 2 ,6-DIALKYL PH E N O L S ! • i t ; F erric ch lo rid e hezahydrate in h o t g la c ia l a c e tic acid was . used in an attem pt to o x id iz e 2 ,6 -d i-t-b u ty lp h e n o l, 2 ,6 -d iiso p r o p y l- I p h en ol, and 2 ,6-diraethylphenol to th e ir r e sp e c tiv e quinones. The i ! ' j products o f t h is o x id a tio n were red , n ot y ello w as would be p red icted j i < j fo r sim ple q uinones. The red oxid ation products were not separable ; j I 1 in to sim pler components by such procedures as fr a c tio n a l c r y s ta l- j ' i l i z a t i o n , vacuum su b lim a tio n , column chromatography, and variou s j ! ' chem ical r e a c tio n s . Further, when the red u ction products o f o x id ized ! ; ' * - 2 ,6 -d iiso p r o p y l- and 2 ,6 -d i-t-b u ty lp h e n o l were added to the pyridinium ; ; i 1 to s y la te o f 111- 2 3 , coupling f a ile d to take p la c e , thus in d ica tin g I | the absence o f an unhindered hydroxyl group. Repeated elem ental i 1 an alyses o f the o x id a tio n product o f 2 ,6 -d i-t-b u ty lp h e n o l, and o f t h is ! product a fte r c a t a ly t ic red u ctio n , f a ile d to confirm the sim ple i I quinone and hydroquinone stru ctu res p o stu la te d . , j 1 1 Hoffmann (5 6 ), in an attempt to o x id ize p y r o g a llo l- l,3 - d i- i i ! m ethyl eth er to i t s quinone, prepared IV-1 using potassium dichromate j i in g la c ia l a c e tic a c id . I ocM -b o Of/ I ! ocfl3 octi 3 0/t/ o Erdtm an ( I4 J 4 ) j, in t h e c o u r s e o f i n v e s t i g a t i o n s on hum ic a c i d . 2-methoxyhydroquinone. Erdtman e sta b lish e d the stru ctu re o f the product as IV -2. A th ir d type o f product m ight be expected from t h is r e a c tio n . The potassium ferricy a n id e o x id a tio n o f 2-n apth ol in d ilu te base g iv es a low y ie ld o f c o lo r le s s dehydro-B-napthol, IV -3, although aqueous f e r r ic g iv es IV-it, a lso c o lo r le s s . A survey o f the a v a ila b le lite r a tu r e revealed th a t the chromic acid o x id a tio n o f 2 ,6 -d i-t-b u ty lp h e n o l in a c e tic acid rep orted ly gave IV-5 was a ssig n ed . The m eltin g p o in t fo r the f e r r ic ch lo rid e oxida tio n product was 2 i|0 -5 °, a fte r two r e c r y s ta lliz a t io n from benzene/ eth an olj 1 /5 . in v e stig a te d the f e r r ic ch lorid e and chromic acid o x id a tio n s o f a red-brown substance m elting a t 21*0° (57)* to which the stru ctu re h 3 The reduction product o f IV -5, to which the stru ctu re IV-6 may be a ssig n ed , was found to have thyromim etic a c t i v it y . This asp ect w i l l be d iscu ssed in Chapter V. The a n a ly tic a l d a ta , found a t the end o f t h is s e c tio n , f a i l to agree w e ll w ith the stru ctu re s proposed, so th a t the q u estio n a r is e s as to whether the diphenohydroquinone, IV -6, or some contam in an t i s resp o n sib le fo r the observed thyromim etie a c t i v it y . As a fu rth er a id in t h is problem , the in frared sp ectra o f IV-5 and IV-6 tio n fo r IV-5 i s not p r e se n t. O therw ise, the sp e ctra are not incom p a tib le w ith the stru ctu res a ssig n ed . The absence o f ab sorption in the 8 .0 -9 .0 u . region argues a g a in st the presence o f an e th e r . The f e r r ic ch lo rid e oxid ation product o f 2 ,6 -d i-i-p ro p y lp h en o l which f a i l s to c r y s t a lliz e . Upon red u ction i t g iv e s , lik e IV -5 , a c o lo r le s s product to which the stru ctu re IV-7 may be te n ta tiv e ly a ssig n ed . are recorded in Table I . I t i s noted th a t a t y p ic a l carbonyl absorp- has n ot been reported to the author’s knowledge. I t i s a red o i l C-tk, c r fj TABEE I I INFRARED ABSORPTION SPECTRA3, Red O xid'n. Product*3 (IV -5) Reduced P roduct Microns R e l. I n t . Microns R e l. 3 .1 £ k 3.1*5 1 * 5.71* 3 « ■ * «n 6 .2 5 12 tm tm 7 .3 0 5 7 .9 8 6 9.17 1 * 9 .6 0 1 .5 11.10 2 5.71* 3 6 .0 5 2 ■» •» 6.98 I * 7 .3 0 3 7.60 1 7 .9 8 6 8 .62 3 8 .9 3 3 «m tm tm • » • » tm m> mm i i . l a i 11.30 a S o lv e n t a b s o r p ti o n h a v e b e e n o m itte d . b S o lv e n t- c a r b o n t e t r a c h l o r i d e . ’ ( I V -6 ) I n t . y EXPERIMENTAL | 3>5?3! ,S l -TETRA-t-BUTYL-k,k' -D IPH EM G Q U INO NE I j . 3 ,5 a 3 1 *5* -T e tr a -t-b u ty l-k 3 h' -diphenoquinone, IV -5. One i : hundred m l. o f hot g la c ia l a c e tic a cid was used to d iss o lv e 2 0 .6 g . (0 .1 m o l.) o f 2 ,6 -d i-t-b u ty lp h e n o l. 7 1 .2 g . (O.k m o l.) o f f e r r ie ch lorid e hexahydrate was added and the m ixture heated on the steam bath fo r one hour w ith o cc a sio n a l shaking. Then 20 m l. o f d i s t i l l e d water was added and the m ixture p laced in the steam cab in et overnight i a t 5 8 °. A fter the r ea ctio n v e s s e l had co o led , 5© ml o f w ater was ; added and the m ixture ex tra cte d w ith benzene. The benzene e x tr a c ts ■were combined, and upon the ad d itio n o f 2 ~k volumes o f 9 5 % eth an ol I * | a mass o f dark m e ta llic -r e d n eed les appeared. A fter r e fr ig e r a tio n I |o v e r n ig h t, the c r y s ta llin e mass was f ilt e r e d o f f and r e c r y s ta lliz e d j | from benzene/ethanolj l / i u M elting p o in t, 2k©-!?0 . The m e ta llic -r e d I j c o lo r tended to disappear on exposure to a i r , lea v in g a brownish c a st | to the product. Y ield yi%, i I i 3 s5 , 3 ' ,5 l -TETRA~t-BUTYL-k,V-DIPHENGHYDRGQUINONE i j I 3 s 5 i 3 ' -T e tr a -t-b u ty l-k , h 1-diphenohydroquinone. IV -6. A I ' ‘ - , i 950 rag. sample o f IV-5 was d isso lv e d in 15 m l. o f g la c ia l a c e tic acid 1 / The an alyses reported here were performed by Mr. W.J. Schenck o f t h is Laboratory, and are recorded a t the end o f t h is s e c tio n . M elting p o in ts are u n corrected . 1 * 6 w ith warning, and then added to a m eg n etica lly s tir r e d suspension j o f one g . o f 10% p a lla d ize d ehareoal in 25 m l. o f g la c ia l a c e tic | ! - I | a c id . The hydrogenation was conducted a t atm ospheric pressure fo r j > 1 * 8 h ou rs, a f te r which the rea ctio n m ixture was warmed, f i l t e r e d , , j ! 1 j and the resid u e washed w ith two sm all p ortion s o f h ot g la c ia l a c e tic ; | i a c id . Evaporation o f the g la c ia l a c e t ic a cid s o lu tio n and recry st~ j a lliz a t io n o f the resid u e from 95% eth a n o l gave a p a le yellow product j appearing as n e e d le s. Y ie ld , 810 mg. (86%), M elting p o in t, I8l*-I8ii.5°- ! i i i I 3 ,5 ,3 s , 5 s -TETRA-i-PROPYl-li, 1 * * -D IPH EN O H Y D R O Q U IN O N E | ' s I 3 ,5 ,3 s , 5 s -T etra -i-p r o p y l-lt, 1 * 1 -diphenohydroquinone. IV- 7 . j i One hundred ml., o f h ot g la c ia l a c e tic acid were used to d isso lv e I 1 7 .8 g . (0 .1 m o l.) o f 2 ,6 -d i-i-p r o p y lp h e n o l. 7 1 .2 g . (O .iim o l.) o f f e r r ic ch lo rid e hexahydrate was then added and the m ixture was heated on the steam bath fo r one hour w ith o cca sio n a l s w ir lin g , then 20 m l. o f w ater added and the m ixture placed in a steam cab in et overnight a t 5 8 °. E xtraction o f the cooled inixture w ith benzene a fte r the i " ; • a d d itio n o f 5© m l. o f d i s t i l l e d w ater, follow ed by evaporation o f the I . 1 j benzene, y ie ld e d a red o i l which f a ile d to c r y s t a lliz e from e th a n o l. j j This o i l i s presumably 3 ,5 ,3 ' ,5* -te tr a -i-p r o p y l-i* ,k #~diphenoquinone. ' I t was d isso lv e d in g la c ia l a c e tic a cid and reduced with hydrogen i I over 10% p a lla d iz ed ch a rco a l. The hydrogenation m ixture was f il t e r e d , ’ ' i I and e x cess a c e tic acid a sp irated on the steam b a th . The so lu tio n was j re fr ig e r a te d fo r two weeks during which tim e d i s t i l l e d water was added dropwise from tim e to tim e, y ie ld in g c lu s te r s o f w hite need l e s . Y ield 20% o v e r a ll from the p h en ol. M elting p o in t, 10l*-5° f table i n ANALYTICAL DATA Compound C a l c u la t e d C H 3 ,5 ,3 * ,5 * -T e tr a - 82.3 9 .8 t-b u ty l-A ,A 8 - d i- phenoquinone (IV -5) 3 ,5 ,3 * ,5***Tetra~ 81.9 1 0 .3 t-butyl~A ,ii* - d l- phenohydroquinone (IV -6 ) 3 ,5 ,3 * ,5 ’ -T etra - 8 l.it 9 .6 i-p r o p y l-itjii8 - d i- phenohydroquinone (IV -7) Found € H 81 .5 9.7 80.8 9.7 CH A PTER V | | ! PRELIM INARY BIOASSAY O F A N A L O G S 1 j I i * ' i ■ • ! The assay procedure used in t h is Laboratory has been amply j d iscu ssed in p u b lica tio n (2 7 ) and in the D octoral D is se r ta tio n ©f j i I T.C. Bruice (1*9)* This b u llfr o g tad pole a ssa y , using Rana O atesbeiana,‘ ■" — — — — — — j f i s su p erior to oth er amphibian assay procedures in so fa r as the t e s t j I i animal i s com m ercially a v a ila b le , low in c o s t , a v a ila b le throughout j the y e a r , easy to handle and measure due to i t s la rg e s i z e , and j i 1 s u f f ic ie n t ly v ia b le to w ithstand repeated measurements. \ Assay Procedure. ! i i S o lu tio n s o f thyroxine and the analog to be te s te d were p re- | j ! pared by d isso lv in g amounts weighed to the n ea rest ten th -m illigram \ ! | in water w ith the a id o f potassium carbonate, and then making d ilu - j | tio n s to obtain the variou s con cen tration ranges d e sir e d . The sto ck I I so lu tio n was added to the t e s t co n ta in ers by means o f graduated j i i i p ip e t t e s , or e ls e an e n tir e volume fo r a given con cen tration le v e l | ; [ j was made up a t one tim e in a la r g e beaker. In e ith e r case the t e s t j I ' | ; so lu tio n s were o f 300 m l. each , and were contained in c le a r p la s t ic I 1 d is h e s , and were o f pH 7 .0 - 7 .5 . The pH adjustment was made w ith po- ! tassium carbonate and h yd roch loric a c id . Groups o f tad p oles w ith ! I t a i l - f i n s measuring 1 .0 - 2 .0 cm. were p laced in in d iv id u a l c o n ta in e r s, I w ith f iv e to e ig h t animals being assayed a t each dose l e v e l . A co n tro l : group o f lik e s iz e was a ls o used in each determ in ation . The anim als i were then placed in a la rg e a ir therm ostat, or in a la r g e , clo sed i ; room o f very n early con stan t tem perature. For the assays rep o rted , the temperature v a ried from 2l*-7°. The tad p oles remained in the h or- ! mone so lu tio n s fo r fo r ty -e ig h t h ou rs, a t which tim e they were in d i- i v id u a lly rin sed and placed in 300 m l. o f tap -w ater. A fter two more i j d ays, they again were p laced in 300 m l. o f tap -w ater. Zero tim e o f j the measurements was taken as the tim e a t which the tad poles were I placed in the hormone s o lu tio n . Measurements were made every tw enty- fou r h ou rs, fo r a t o t a l o f s i x d ays. The a c tu a l measurements o f the t a i l - f i n width were estim ated to w ith in tw o-ten th s o f a m illim e te r . I n i t i a l l y th ere i s a prelim inary change in t a i l - f i n width ; during the f i r s t tw enty-four to fo r ty -e ig h t h ou rs, a fte r which the average group response becomes e s s e n t ia lly lin e a r . A d e ta ile d p res en ta tio n o f B ru ice’s method (27, ii9) o f a ssig n in g a c t iv it y va lu es w i l l | not be g iv en . j The assays reported here are e n t ir e ly approximate and p relim - | in a ry , and in e f f e c t serve to in d ic a te the appropriate con cen tration i ranges to be used in fu tu re t e s t in g , rath er than the a c tu a l f in a l ' assay valu es fo r the su b stan ces. 1 TABLE IV ACTIVITIES OF THYROXINE A N A LO G S AS DETERMINED B Y y THE RA N A CATESBEIANA TAIL-FIN ASSAY COMPOUND u g ./m l. O > (V0 )/c o n c . in y / m l. 3 / A c tiv ity V L -th y ro x in e (1 -1 ) 3 ,5 -D iio d o -d e s - am inothyronine 1.0123 3-036? 5.0615 0 .0 9 0 .2 5 o . b o 0.0885 0.0810 0.0803 200$ 1 .0 6 6 0.17 0.159b c a . b00$ E th y l 3 ,5 - d i n i t r o - b - ( 2 ’ ,5 ’ -dim eth o x y - b '-h y d ro x y p h en o x y ) - hydrocinnam ate ( I I I - l l ) 1.0 0 2 .0 0 0 .0 1 3 0.0066 c a . 10- 30$ y v e ry h ig h 800-5000$ 3 ,5 -D iio d o -b - ( 2 1, 5 ' -dim ethoxy- b ' -hydroxyphenoxy) - hydrocinnam ic Acid ( I I I - 1 5 ) 0 .1 0 6 6 2 .0 0 o . o b 0 .3 b 5 \ V 3 , 5 , 3 ', 5 '- t e t r a - t - b u t y l - b , b '- d i - phenohydroquinone 0.b3k8 b.838 0 .0 3 0.0620 c a . 60$ " ^ to x ic I. / The a u th o r i s in d e b te d to M iss M arg aret K ulik o f t h i s L ab o rato ry who perforated m ost o f th e a s s a y s . 2 / P e r-c e n t d e c re a se p e r hour a t 2 b -7 ° . 3 / P e r-c e n t a c t i v i t y a d ju s te d to a m olar b a s is a c c e p tin g th e L -th y ro x in e s ta n d a rd as 200$, \x / K indly fu rn is h e d as th e sodium s a l t by T rav en o l L a b o ra to rie s , I n c . , M orton G rove, I l l i n o i s . $ / T his analo g has ro u g h ly 200-300$ a c t i v i t y d u rin g th e f i r s t 2-3 d a y s. I t s f u r t h e r d e c lin e in a c tio n su g g e sts an a c t i v i t y a k in to t h a t o f 2 , L - d in itro p h e n o l. 6 / T his a c t i v i t y f ig u r e i s m ean in g less as th e io d in e a n a ly s is o f th e t e s t s compound proved to be v e ry low , in d ic a tin g c o n ta m in a tio n by in o rg a n ic s a l t s . 7 / T h is an alo g o f d ip h e n y l i s v e ry p o o rly s o lu b le in c o n c e n tra te d b ase as w e ll as a t pH 7 - 7 .5 . 8 / T his an alo g was p ro v id ed by P ro fe s s o r Norman K h arasch . CHAPTER VI l i S U M M A R Y A N D C O N C L U SIO N S i \ The purpose of th is in v estig a tio n was to prepare and t e s t a se le c te d group of a lk y l and alkoxy analogs of thyroxine. ■ The f i r s t phase was concerned with the preparation of a number ■of a lk y l-su b stitu ted phenols and hydroquinones which could be coupled iwith e th y l 3 ,5 -d in itro p h lo reta te so as to give a large se r ie s of d i- initrodiphenyl e th er s. The second phase was the conversion o f these dinitrodiphenyl eth ers in to th e ir resp ective diiodo analogs. A third phase con sisted of te stin g -th e analogs so as to further jdefine the stru ctu ral lim its fo r thyromimetic substances as an aid to 1 ' ifuture in v estig a tio n s in to the mechanism of th yroxin e-lik e a c tiv it y . ! A sid e lig h t which developed during the course of the work was jthe observed thyroraimetie a c tiv ity of a su b stitu ted diphenohydroquinone I f v e r ifie d , th is observation i s of sp e c ia l in te r e st with regard to the i ■earlier hypothesis of Niemann and coworkers on the mechanism o f thy- jroxine a etio n . Only the fir st'p h a se of th is undertaking has been reasonably jeompleted. The iod in ation and bioassay of the remaining compounds i iremains fo r future work. BIBLIOGRAPHY BIBLIOGRAPHY I 1 . C oindet, C.W. Rev. med. h i s t , e t p h il. P a r is , 1821. 2 . Baumann, E. and H, Roos. Z. p h y sio l. Chem. , 21 , k8l (1 8 9 6 ). i 3 . Oswald, A. Z. p h y sio l* Chem. 2 7 , lit (1 8 9 9 ). i ! k . M arine, D. and C.H. Lenhart. Arch. I n t . Med. k, 2$3 (1 9 0 9 ). i 5 . K endall, E.G. Tr. A ss. Am. P h y sicia n s. 30? k20 (1 9 1 $ ). 6 . H arington, (G.R. Bioehem. J . 21, 169 (1 9 2 7 ). 7 . S a lte r , W .T., J . Lerman, and J.H . Means. Jour. C lin . In v e stig a tio n . I k , 37 (1935)* 1 8 . H arington, C.R. E ndocrinol. k9, k01 (1 9 5 1 ). i i ! i 9 . Johnson, T.B. and L .B . Tewkesbury, J r . Proc. N at. Acad. j S c i. U .S.A . 28, 73 <19k2). ! ‘ ! 1 0 . H arington, C.R. J . Chem. S oc. 193 (I9 k k ). t f 1 1 . Neuberger, A. Ann. Rev. Bioehem. 18 , 2k3 (I9 k 9 ). 1 2 . H arington, C.R. B r it . Med. Jour. 1269 I I (1 9 3 6 ). 1 3 . G ross, J* and R. P itt-R iv e r s . Lancet 262, 593 (1 9 5 2 ). i '■ i ■Ik. G ross, J . and R. P itt-R iv e r s . Bioehem. J . 53? 652 (1953)* i i t |15. S a lte r , W.T. A Textbook o f Pharmacology. W.B. Saunders and Co. ! P h ila d elp h ia , 1952. " | jl6 . Selenkow, H.A. and S .P . Asper, J r . P h y sio l. R evs. 35 , k26 (1955)* ! |1 7 - F ried en , E. and R .J . W inzler. Jour. B io l. Chem. 176, 155 (I9 k 8 ). | ;18. C an za n elli, A. and D. Rapport. Am. J . P h y sio l. 103, 279 (1933)• 1 \ |19. Roche, J . , S . l i s s i t z k y and R. M ich el. Gampt. rend. Acad. s c . j 23k, 1228 (1 9 5 2 ). 1 120. Niemann, C. F o r ts, der Chemie o rg . Natur. 7 , 173 (1 9 5 0 ). j . } j21. H arington, C.R. F o r ts, der Chemie org. Natur. 2 , 121 (1939)* j 55 22 . Niemann, C . and J . F . Mead J . Am. Chem. S oc. 6 3 , 2685 (19i*l). 23 . Niemann, G. and G.E. Redemann J . Am. Chem. S oc. 6 3 , 151*9 (191*1). 21*. Boyer, P.B . Proc Soc .Exp. B io l. Med. 1*9, 176 (191*2). 25* B ru ice, T.C. J . Org. Chem. 1 9 , 333 (1951*). 2 6 . Kharasch, N. J . Chem. Ed. 32 , 192 (1 9 5 5 ). 27 . B ru ice, T X ., R .J . W inzler, and N. Kharasch. J . B io l. Chem. 210, 1 (1951*). 28. L oeser, A ., H. Ruland and V.M. T rik o ju s. Naunyn-Schmiedebergs Arch, exp . Path. Pharraakol. 189, 661* (1 9 3 8 ). 29. C layton , J .C . and B.A. Hems. J . Chem. S oc. (1950) 81*0. 30 . H arington, C.R. Bioehem. J . 1*3, i*3l* (191*8). 31 . M yers, C .S . J . Am. Chem. S oc. 51*, 3818 (1 9 3 2 ). 32 . C layton, J .C ., G.F.H. Green and B.A. Hems. J . Chem. Soc. (1951) 21*69. 33 . B ru ice, T .G ., N. Kharasch and R .J . W inzler. J:. Org. Chem. 1 8 , 83 ( 1 9 5 3 ) .’ ~ 31*. von Konek, F . , and E. Pascu. B er. 5 1 , 859 (1 9 1 8 ). 35* Schwenk, E . and B . Papa. J . Org. Chem. 10 , 232 (191*5). 36. Hanke, M.T. and K.K. K o essler. J . B io l. Chem. 50 , 21*5 (1 9 2 2 ). 37. B r u ic e , T X . J . O rg . Chem. 1 9 , 333 (1951*). 3 8 . C allow , R .G ., N.M. Gulland and R.D. Haworth. J . Chem. S o c. (1929) 11*52. 3 9 . F i s c h e r , E . and A. S p e i e r . B er* 28, 3253 (1 8 9 5 ). 1*0. Baker, A. and N X . Brown, J , Chem. S o c. (191*8) 2303. 1*1. C lark, T.H. Am. Chem. Journal .U *» 565 (1892). 1*2. F ie s e r , L .F . Experiments in Org. Chem. Page 229. D.(G. Heath and Co. N.Y. 19l*l. 56 1*3. N o e ltin g , E. and S . F o r e l. B er. 1 8 , 2673 (1 8 8 $ ). 1 * 1 * . Erdtman, H.G.H. Proe. Roy. S oc. (London). All*3, 191 (1 9 5 3 ). 1*5. Bamberger, E . and J . F r e i. B er. 1*0, 191*1* (1 9 0 7 ). 1*6. Rugheimer, L. and M « H ankel. B er. 2 9 , 2171* (1 8 9 6 ). 1*7. K noevenagel, E .a n d C . B uckel. B er. 31*, 3993 (1 9 0 1 ). 1*8. F ie s e r , L .F. Experiments in Org. Chem. Pages 171 and 181. D.C. Heath and Co. N.I. 19l*l. 1*9. B ru ice, T.C,, D is s e r ta tio n , U n iv ersity o f Southern C a lifo r n ia , June, 1951*. 5 0 . Ullmann, F . and G. N adai. Ber. 1*1, 1822 (1 9 0 8 ). 51 . Niemann, C . and G.E. McGasland. J . Am. Chem. S oc. 6 6 , 1870 (191*1*). — 52. Baker, ¥ . and R.I. Savage. J . Chem. S o c. (1938) 1602. 53• Mauthner, F . J . p r. Chem. 89, 30l* (1911*). 51*. Baker, W . and H.A. Sm ith. J . Chem. S o c. (1931) 251*2. 55 . H erzig , J . and J . P o lia k . B er. 3 6 , 661 (1 9 0 3 ). 5 6. Hoffman, A.W. B er. 1 1 , 337 (1 8 7 8 ). 57 . B r it . P a t. 686, 261, reported in Chem. Abs. 1*8, l*593b (1951*). (S a lv ers! tv o f Southern C a lifo r s lfe

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Core Title The synthesis and testing of some alkyl substituted thyroxine analogs

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