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UBC Theses and Dissertations

The synthesis of some antergan analogues with unsaturated cyclohexyl and substituted aromatic rings Park, Jung Kil 1974

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T H E SYNTHESIS O F S O M E A N T E R G A N A N A L O G U E S WITH U N S A T U R A T E D C Y C L O H E X Y L A N D S U B S T I T U T E D A R O M A T I C RINGS by J U N G K I L P A R K B . S c . in Pharmacy, The Seoul National University , 1965 A THESIS S U B M I T T E D IN P A R T I A L F U L F I L L M E N T O F T H E R E Q U I R E M E N T S F O R T H E D E G R E E O F M A S T E R O F S C I E N C E ( i N P H A R M A C Y ) in the Division of Medicinal Chemistry of The Faculty of Pharmaceutical Sciences We accept this thesis as conforming to the required standard T H E U N I V E R S I T Y O F BRITISH C O L U M B I A January, 1974 In p r e s e n t i n g t h i s t h e s i s in p a r t i a l f u l f i l m e n t o f the r e q u i r e m e n t s f o r an advanced degree at the U n i v e r s i t y o f B r i t i s h Co lumb ia , I a g ree that the L i b r a r y s h a l l make i t f r e e l y a v a i l a b l e f o r r e f e r e n c e and s tudy . I f u r t h e r agree t h a t p e r m i s s i o n f o r e x t e n s i v e c o p y i n g o f t h i s t h e s i s f o r s c h o l a r l y purposes may be g r a n t e d by the Head o f my Department o r by h i s r e p r e s e n t a t i v e s . It i s u n d e r s t o o d that c o p y i n g o r p u b l i c a t i o n o f t h i s t h e s i s f o r f i n a n c i a l g a i n s h a l l not be a l l o w e d w i thou t my w r i t t e n p e r m i s s i o n . The U n i v e r s i t y o f B r i t i s h Co lumbia Vancouver 8, Canada Date A B S T R A C T Ant e r g a n i s one of the ethylenediamine type of a n t i -h i stamines; in this work ten analogues of it were p r e p a r e d . That p ^ conjugation i s e s s e n t i a l to antergan's a n t i h i s t a m i n i c a c t i v i t y has a l r e a d y been established. Six of these analogues ( A - l a-d, and B - l and 2, i l l u s t r a t e d i n F i g . £ , p. 4 ) were synthesized in o r d e r that the s i g n i f i c a n c e of p;& conjugation in the a n t i h i s t a m i n i c p r o p e r t i e s of antergan m o l e c u l e may at some future time be investigated. The p r o b l e m is to find i n what way a l t e r a t i o n ( i . e. , i n c r e a s e or d e c r e a s e in) e l e c t r o n i c density of the p/fr -conjugated moiety of the m o l e c u l e affects a n t i h i s t a m i n i c a c t i v i t y . The analogues A - l a-d i n v o l v e d ortho-, meta-, and p a r a - m e t h y l , and p a r a - b r o m o substitution to the a r o m a t i c r i n g which gives r i s e to p^j conjugation in the antergan molecule; while the r i n g which gives r i s e to homo conjugation was r e p l a c e d by a c y c l o h e x y l moiety to eliminate any p o s s i b l e contribution of homoconju-gation to a n t i h i s t a m i n i c a c t i v i t y . In analogue B - l , the antergan s t r u c t u r e was m o d i f i e d so that the a r o m a t i c r i n g which gives r i s e to p/£, conju-gation was r e m o v e d f r o m the r e s t of the m o l e c u l e by a methylene group, i . e. , the phenyl group was r e p l a c e d by a b e n z y l moiety. The other r i n g was left unmodified. Another compound (B-2) r e l o c a t e d the a r o m a t i c r i n g giving r i s e to p ^ conjugation to the adjacent methylene carbon, so that p ^ conjugation was eliminated; this compound i s the i i nitrogen analogue of diphenhydramine and thiodiphenhydramine. In order that the importance of homo conjugation to antergan's antihistaminic activity may be established, the aromatic ring which gives rise to p^, conjugation was replaced by a cyclohexyl moiety (A-3). Two compounds were synthesized in which the aromatic ring giving rise to homoconjugation were replaced by 3-cyclohexyl moieties (A-2a and b); while the aromatic ring which gives rise to pjij, conjugation was left unaltered in one of the compounds (b) and replaced by a cyclohexyl moeity in the other (a). In the tenth compound (A-1 e), both aromatic rings giving rise to both homo- and pfij. - conjugation were removed and replaced by cyclohexyl moieties. The resulting analogue of antergan has already been demonstrated to have a very low antihistaminic activity compared to diphenhydramine, but it was felt that it would provide a useful com-parison for the antihistaminic activities of the other antergan analogues prepared in this work. Two final intermediates in the synthesis of other analogues of antergan were prepared. In these analogues (C-2a and b) the aromatic ring giving rise to homoconjugation would have been replaced by the 1-cyclohexenyl moiety, while the other aromatic ring which gives rise to pfo conjugation would have been the same in one of the analogues, and replaced by a cyclohexyl moiety in the other. ii i In intermediates of analogues D - l and 2, the ring giving rise to homoconjugation would have been replaced by 2, 5- and 1,4-cyclohexadiene respectively, while the aromatic r ing giving r ise to P7L conjugation was replaced by the cyclohexyl moiety. In intermediates D-3 and 4, the aromatic ring giving r ise to homoconjugation would have been replaced by cyclohexyl, while the aromatic ring giving r ise to p H, conjugation would have been replaced by 2, 6-dimethylphenyl and 3-cyclohexenyl moieties. iv A C K N O W L E D G E M E N T I express my most sincere thanks to D r . T . H . Brown for his understanding, and help in resolving technical difficulties encountered in this work. I also wish to extend my appreciation to Dean B . E . Riedel of the Faculty of Pharmaceutical Sciences for his understanding, and inspiration. F inancial support f rom the Universi ty of B r i t i s h Columbia is gratefully acknowledged. v T A B L E O F C O N T E N T S Page A B S T R A C T i i A C K N O W L E D G E M E N T v T A B L E O F C O N T E N T S vi LIST O F F I G U R E S xi i LIST O F T A B L E S xiv. I. I N T R O D U C T I O N 1 II. DISCUSSION O F T H E C H E M I S T R Y 3 III. A N A L Y T I C A L M E T H O D S 46 IV. E X P E R I M E N T A L 47 A . SYNTHESIS O F N , N - D I M E T H Y L - N ' - C Y C L O -H E X Y L M E T H Y L - N ' - o - M E T H Y L P H E N Y L -E T H Y L E N E D I A M I N E 47 1. <k -Chloro-o-methylphenylacetamide 47 2. o( -Dimethylamino-N-o-methylphenyl -acetamide 48 3. . N , N - dim e thy I - N 1 - o - m ethyIpheny 1 -enediamine 49 4. . N / - £ - m e t h y l p h e n y l - N - ( / ? - N , N - d i m e t h y l -aminoethyl)- cyclohexanecarboxamide 51 5. N , N-dimethyl -N' -cyclohexylmethy 1 - N 1 -o-methylphenylethylenediamine 52 B . N , N - D I M E T H Y L - N ' - C Y C L O H E X Y L M E T H Y L -N ' - m - M E T H Y L P H E N Y L E T H Y L E N E D I A M I N E . . 54 1. o ( - C h l o r o - N -m-methylphenylacetamide . . . 54 2. fj\ - D i m e thyl a m i n o - N - m - m e thy Ipheny 1 -acetamide 55 3. N , N-dimethyl -N' -m-methvlphenyle thvl -enediamine 57 vi T A B L E O F C O N T E N T S (Continued) Page B . N , N - D I M E T H Y L - N ' - C Y C L O H E X E Y L M E T H Y L -N 1 - m - M E T HY L P H E N Y L E T HY L E N E D I A M I N E (Continued) 4. . N ' - m - m e t h y l p h e n y l - N ' - ^ - N , N - d i m e t h y l -aminoethyl)-cyclohexanecarboxamide 58 5. . N , N - d i m e t h y l - N ' - c y c l o h e x y l m e t h y l - N 1 -m-methylphenylethylenediamine 60 C . . N , N - D I M E T H Y L - N ' - C Y C L O H E X Y L M E T H Y L -N ' - p - M E T H Y L P H E N Y L E T H Y L E N E D I A M I N E 61 1. o^-Chloro-N-p-methylphenylacetamide 61 2. e< -Dimethylamino-N-p-methylphenyl -acetamide 62 3. N , N-dimethyl -N' -p-methylphenylethyl -ene diamine 63 4. N ' - p - m e t h y l p h e n y l - N - ( / - N , N - d i m e t h y l -aminoethyl)-cyclohexanecarboxamide 65 5. N , N - d i m e t h y l - N ' - c y c l o h e x y l m e t h y l - N 1 -p-methylphenylethylenediamine 65a D . . S Y N T H E S I S O F N , N - D I M E T H Y L - N ' - C Y C L O -H E X Y L M E T HY L - N ' - p- B R O M O P H E N Y L -E T H Y L E N E D I A M I N E 66 1. <k-Chloro-N-p-bromophenylacetamide 66 2. o( -Dimethylamino-N-p-bromophenyl-acetamide 67 3. N , N-dimethyl -N' -p-bromophenyle thyl -enediamine 68 4. . N - p - b r o m o p h e n y l - N - ( ft - N , N - d i m e t h y l -aminoethyl)-cyclohexanecarboxamide 70 5. N , N - d i m e t h y l - N ' - c y c l o h e x y l m e t h y l - N ' -p-bromophenylethylenediamine 71 E . -SYNTHESIS O F N , N - D I M E T H Y L - N ' - C Y C L O -H E X Y L M E T H Y L - 3 , 5 - D I M E T HY L P H E N Y L -E T H Y L E N E D I A M I N E 72 1. c / - C h l o r o - N - 3 , 5-dimethylphenylacetamide . . 72 2. o ( -Dimethylamino-N-3 , 5-dimethylphenyl-acetamide 74 3. . N , N - d i m e t h y l - N ' - 3 , 5-dimethylphenyl-ethylenediamine 75 vi i T A B L E O F C O N T E N T S (Continued) Pa-ge E . SYNTHESIS O F N , N - D I M E T H Y L - N ' - C Y C L O -H E X Y L M E T H Y L - 3 , 5 - D I M E T H Y L P H E N Y L -E T H Y L E N E D I A M I N E (Continued) 4. N - 3 , 5-dimethylphenyl-N-( fi - N , N-dimethyl -amino)- eye lohexane c arboxamide 76 F . . N , N - D I M E T H Y L - N 1 - C Y C L O H E X Y L M E T H Y L r N 1 - CY C L O H E X Y L E T H Y L E N E D I A M I N E 78 1. Cyclohexanecarboxyl chloride 78 2. d -Chloro-N-cyclohexylacetamide 78 3. J. -Dimethylamino-N-cyclohexylacetamide . . . 80 4. N , N-dimethyl-N' -cyclohexylethylene-diamine 81 5. N - c y c l o h e x y l - N - ( ^ - N , N-dimethylamino-ethyl)-cyclohexanecarboxamide 82 6. N , N - d i m e t h y l - N ' - c y c l o h e x y l m e t h y l - N 1 -cyclohexylethylenediamine 83 G . -SYNTHESIS O F N , N - D I M E T H Y L - N ' - B E N Z Y L -N ' - C Y C L O H E X Y L E T H Y L E N E D I A M I N E 85 1. N - c y c l o h e x y l - N - ( ^ - N , N-dimethylamino-ethyl)-phenylcarboxamide 85 2. N , N - d i m e t h y l - N ' - b e n z y l - N ' - c y c l o h e x y l -ethylenediamine . . . 86 H . N , N - D I M E T H Y L - N - D I P H E N Y L M E T H Y L -E T H Y L E N E D I A M I N E 87 1. <A -Chloro-N-diphenylmethylacetamide 87 2. c>i -dimethylamino-N-diphenylmethyl-acetamide 89 3. N , N-dimethyl-N'-diphenylmethylethylene-di amine 90 I. N , N - D I M E T H Y L - N ' N ' - D I B E N Z Y L E T H Y L E N E -D I A M I N E 91 1. N ' - b e n z y l - N ' - ^ - N , N-dimethylaminoethyl)-benzenecarboxamide 91 2. N , N - d i m e t h y l - N ' , N'-dibenzylethylene-diamine 92 vi i i T A B L E O F C O N T E N T S (Continued) Page J . N , N - D I M E T H Y L - N 1 - 1 - C Y C L O H E X E N Y L -M E T HY L - N C Y C L O H E X Y L E T HY L E N E -D I A M I N E 94 1. . Cyclohexylcyanohydrin 94 2. Methyl 1-hydroxy-cyclohexanecarboxylate . . . 95 3. Methyl 1-cyclohexenecarboxylate 96 4. 1-cyclohexenecarboxylic acid 97 5. 1-cyclohexenecarboxyl chloride 98 6. . N-cyc lohexyl -N- ( ^ - N , N-dimethylamino-ethyl)-1-cyclohexenecarboxamide 98 K . N , N - D I M E T HY L - N 1 - 3- C Y C L O H E X E N Y L -M E T H Y L - N ' - C Y C L O H E X Y L E T HY L E N E -D I A M I N E 99 1. 3-Cyclohexenecarboxyl chloride 99 2. N - cyclohexyl-3-cyclohexenecarboxamide . . . 100 3. . N-cyclohexyl -N-3-cyclohexenylmethyl -amine 101 4. - C h l o r o - N - 3 - c y c l o h e x e n y l m e t h y l - N -cyclohexylacetamide 102 5. r£ -Dimethylamino-N-3-cyc lohexenyl -methyl-N-cyclohexylacetamide 103 6. N , N - D i m e t h y l - N ' - 3 - c y c l o h e x e n y l m e t h y l -N'-cyclohexylethylenediamine 105 L . N , N - D I M E T H Y L - N ' - 3 - CY C L O H E X E N Y L -M E T H Y L - N ' - P H E N Y L E T H Y L E N E D I A M I N E 106 1. Ji - Chloro-N-phenylacetamide 106 2. -Dimethylamino-N-phenylacetamide 107 3. N , N-dimethyl-N'-phenylethylenediamine . . . 108 4. . N ' - p h e n y l - N ' - f ^ - N , N - d i m ethyl amino-ethyl)-3-cyclohexenecarboxamide 109 5. . N , N-dimethyl -N' -3 -cyc lohexenylmethyl -N'-phenylethylenediamine I l l M . N - C Y C L O H E X Y L - 2 , 5 - C Y C L O H E X A D I E N E C A R -B O X A M I D E 112 1. 2, 5-cyclohexadienecarboxylic acid (1,4-dihydrobenzoic acid 112 ix T A B L E O F C O N T E N T S (Continued) Page M . N - C Y C L O H E X Y L - 2 , 5 - C Y C L O H E X A D I E N E C A R -B O X A M I D E 112 (Continued) 2. 2, 5-cyclohexadienecarboxyl chloride . . 113 3. N-cyclohexyl -2 , 5-cyclohexadiene-carboxamide 114 N . - N - 2 , 6 - D I M E T H Y L P H E N Y L - C Y C L O H E X Y L M E T H t L -A M I N E 115 1. . N - 2 , 6-dimethylphenyl-cyclohexane-carboxamide 115 2. N-2 , 6-dimethylphenyl-cyclohexylmethyl-amine 116 O . N , N - D I M E T H Y L - N 1 - C Y C L O H E X Y L M E T H Y L -E T H Y L E N E D I A M I N E 117 1. -dimethylaminoethyl-N-cyclohexane-carboxamide 117 2. N , N-dimethyl -N' -cyclohexylmethylethyl -enediamine 118 P . S I N G L E R E A C T I O N S 119 1. 3 - chloro-4 -cyc lohexene- l , 2-dico;rboxylic acid anhydride 119 2. Cyclohexyl p-toluensulfonate 120 3. 1-Chloro 3*cyclohexene 121 4. 3-Cyclohexenylmethylamine 122 5. o( - C h l o r o - N , N-dimethylacetamide 123 6. ci - C h l o r o - N - 2 , 6 -dimethylphenyl -N-cyclo-hexylmethylacetamide (attempted) 124 7. d.-Chloro-N-diphenylacetamide (attempted) 124 V . H I S T A M I N E 126 VI . H I S T A M I N E " R E C E P T O R S " 159 VII. A N T I H I S T A M I N E 163 x T A B L E O F C O N T E N T S (Continued) Page VIII. C O N C L U S I O N 174 B I B L I O G R A P H Y 178 < xi LIST O F F I G U R E S Figure Page 1 Structures of Diphenhydramine (a) and Antergan (b) 1 2 Illustrating the Various Analogues and Intermediates of Analogues of Antergan Prepared in this Work 4-5 3 Mechanism of Amide Reduction 15 4 Reduction of 0 -Unsaturated Amide with L A H 16 5 Mechanisms for Reaction Between Carboxylic A c i d and Carbodiimide 29 6 Mechanisms of B i r c h Reduction of Benzoic A c i d and Aniline 32 7 Geometr ical Isomers of 1-Chlorobutadiene 36 8 Hypothetical Heparin-Prote in-Histamine Complex of Mast C e l l Granules at pH7 130 9 Degranulation and Histamine Release in Mast C e l l s : Suggested Mechanism of Action of Compound 48/80 133 10 Schematic Drawing to Show the Sequential Exocytosis of Histamine-containing Granules 133 11 The Catabolism of Histamine 135 12 Decarboxylation of Histidine 140 13 Schematic Representation of the Approach of Histamine to the Parietal Cel ls of the Stomach 151 14 Conformations of Histamine 160 xii LIST O F F I G U R E S (Continued) Figure Page 15 Schematic Representation of Histamine Receptor 162 16 General Structural F o r m u l a of A n t i -histamines 164 x i i i LIST O F T A B L E S Table Page I o( -Chloroacetanilide Derivatives 8 II °( - D i m e t h y l a m i n o - N - R ^ - A c e t a m i d e Derivatives 11 III N j N - Dime thyl- N 1 - R ? - Ethylene diamine 12 IV N - R 2 - N ' - ( ^ - N , N-Dimethylaminoethyl)-Cyclohexanecarboxamide 20 V N - R 2 - N - ( - N , N-Dimethylaminoethyl)-3-Cyclohexene 22 VI N , N - D i m e t h y l - N ' - C y c l o h e x y l m e t h y l - N I - R 2 -E'thylenediamines (Analogues of Antergan) 24 VII N , N - D i m e t h y l - N ' - 3 - C y c l o h e x y l m e t h y l -methylethylenediamine (Analogues of Antergan A - 2 a and b) 26 VIII Action of Drugs on the A l l e r g i c Response 158 IX Antihistamines 166 xiy. ' : my Parents with Love xv P A R T I 1 I N T R O D U C T I O N Interactions of lone pair electrons of hetero atoms like nitrogen or oxygen are characteristic of aromatic amines and ethers. The lone pair electrons interact with the electrons of an adjacent double bond or aromatic system, or with a double bond or aromatic system remote f rom the hetero atom by one or more methylene groups ( 1 ). p,-j^  conjugation is said to take place when adjacent systems are i n -volved, and the term homoconjugation is used when the interacting systems are separated by a methylene group. Diphenhydramine has • C H - Q - C H 2 - C H 2 - N - C H 3 C H , Diphenhydramine (a) - C H - N - C H 2 - C H 2 - N - C H 3 C H , Antergan (b) Figure 1. only homoconjugation but antergan has both homoconjugation and p/£, conjugation (Fig . 1 ). It was observed by Nauta _et _al. that alkylsubsti -tution of one or both phenyls in compounds like diphenhydramine s ig-nificantly affects interaction between the lone pair electrons of oxygen and those of substituted phenyl, and a certain degree of coplanarity was observed between one phenyl group and the C - O bond ( 2 , 3 ). They also demonstrated that for the alkylsubstituted analogues of diphenhy-o 2 dramine they synthesized, antihistaminic, anticholinergic and local anaesthetic activities were greatly modified by mesomeric , inductive and steric effects resulting from the alkyl substitution. The presence of both phenyl groups in diphenhydramine was of prime importance in the maintenance of maximum antihistaminic activity, because replace-ment of one or both phenyl groups of diphenhydramine by cyclohexyl groups reduced antihistaminic activity ( 2 , $ ). Final ly , Nauta et a l . proposed that the charge density on the ether oxygen of diphenhydramine type compounds is important in determining their pharmacological activities. Antihistaminic activity is supposed to be related to relatively low electron density on the oxygen. Conversely, anticholinergic activity is associated with relatively high electron density. They also suggested that coplanarity of the C - O bond with a phenyl group results in an overlap interaction and is important in the binding of the phenyl group to the binding site of the receptor. This proposal was supported by studies of a series of thioether analogues of substituted diphenhydramine which showed extremely low antihistaminic activity and a general increase of anticholinergic activity due to dif-ferent electronic and steric properties of sulphur atoms compared to oxygen atoms (4"). Work in our laboratory ( 6 ) showed that the replacement of a phenyl group of antergan by a cyclohexyl group, giving r ise to homo-conjugation, resulted in an antihistaminic activity equivalent to that of 3 diphenhydramine. Replacement of both phenyl groups by cyclohexyl groups resulted in extremely low antihistaminic activity. The object of this research was to synthesize analogues of antergan by replacing one or both of the aromatic rings (R^ and R 2 in F i g . 2) with methyl or halogen-substituted phenyl, or with cyclohexyl, cyclohexenyl, or cyclohexadienyl groups. The structures of the c o m -pounds are shown in F i g . 2. The relationship between the antihist-aminic activities of compounds synthesized in the present work and electronic effects (orbital overlap interactions) resulting f rom homo-conjugation or p / t conjugation in combination with m e s o m e r i c , inductive, and steric effects of substituents remains to be investigated. P A R T II DISCUSSION O F T H E C H E M I S T R Y The present work is concerned with the synthesis of ten analogues of antergan (groups A and B in F i g . 2). In addition, two final intermediates (group C - l and C-2) of other analogues of antergan have been obtained. The synthesis of four more analogues (group D) was attempted, but due to their instability and synthetic difficulties with the intermediates, only the intermediates indicated in the Figure were ob-tained. . Nauta et a l . ( £ , 3 ) reported that alkyl substitution on one or both phenyl groups of diphenhydramine altered the electronic inter-action between p orbital electrons of the oxygen and 77J bond 4 Figure 2 . Illustrating the various analogues and intermediates of analogues of antergan prepared in this work. B R r C H 2 - N - C H 2 - C H 2 - N - C H 3  R 2 C H 3 1 Rl = cyclohexyl = a o-me thy Ipheny 1 b m-methylphenyl c p-methylphenyl d p-bromophenyl e cyclohexyl 2 R i = 3 - c y c i ° n e x e n y i R 2 = a c y c l o h e x y 1 b phenyl 3 R i = P h e n y l R-2 = cyclohexyl R - N - C H - C H - N - C H 1 | 2 2 | 3 R 2 C H 3 1 R^ = benzyl R^ = benzyl 2 R-^  = H R 2 = diphenylmethyl R - N - C H - C H - N - C H 1 | 2 2 | 3 R 2 C H 3 1 R = cyclohexane carbonyl K = 3, 5-dime thy Ipheny 1 • 1 2 R l = l - cydohexenecarbonyl R^ = a phenyl b cyclohexyl Figure 2 . Illustrating the analogues D of (Continued) antergan 'p the synthesis of which was attempted. Only the intermediates l* -4 ' were prepared. R - C H - N - C H , - C H , - N - C H , 1 2 1 2 2 I 3 R, C H , D 1 2 3 4 R = R, = R, R, 2, 5-cyclohexadienyl 1, 4- cyclohexadienyl cyclohexyl cyclohexyl R R, R„ = R, cyclohexyl cyclohexyl 2, 6-dime thy Ipheny 1 3- cyclohexenyl 0V,C/0H C H 2 - M H l 1 2, 5-cyclohexadiene-l -carboxylic acid. 2' 3- chloro-4-cyclohexene-1, 2-dicarboxylic acid anhydride. 3' — 2.' ^ " dimethylphenyl- cyclohexylmethylamine. 4' 1- chloro-3-cyclohexene. 6 electrons of the phenyl groups by inductive, mesomeric , or steric effects, or some combination of these. The charge density on the oxygen, which is affected to a certain extent by overlap or d e r e a l i z a -tion with adjacent groups, was claimed to be a significant factor i n the pharmacological activity of the molecule. Steric considerations in alkylated analogues and conformational aspects like coplanarity between one of the phenyl groups and the oxygen were also considered to be important factors in the pharmacological activity of diphenhydramine. Because of the s imilar structures of antergan and diphenhydramine, it was felt that a s imilar approach to that taken by Nauta et a l . would be useful in establishing the significance of electronic and steric effects in antergan's pharmacological action. Four analogues ( A - l , a -d in F i g . 2 ), and two intermediates ( C - l and D-5 1 ) of antergan were synthesized to ascertain the pharma-cological effects of o ^ m - ^ r p - m e t h y l - , 2,6- or 3, 5 -dimethyl- , and p - b r o m o - substitution on the phenyl group of antergan. pTL conjugation is present in all of these compounds. Analogues (A-3 and B - l and 3) were synthesized p r i m a r i l y to ascertain the pharmacological significance of the aromatic ring which gives r ise to pTC conjugation in antergan and its analogues. In this series, the aromatic ring was replaced either by cyclohexyl (A-3), or it was removed from the nitrogen atom and attached to an adjacent carbon atom (B-3). Replacement of R 2 by a benzyl group (B- l ) may result in less overlap than that seen in the pTC system with phenyl. The analogues of antergan ( A - 2 a and b), a final inter-mediate ( C - 2 ) and intermediates ( D - l 1 and 2 1 ) have a cyclohexenyl or cyclohexadiene group in the place of R^. Because of conformational differences and electronic interaction of the diene or olefinic bond in the s ix -membered ring with the nitrogen atom, it is anticipated that the pharmacological activity of these compounds wil l be different in intensity f rom that of antergan. R E A C T I O N O F P R I M A R Y A M I N E S W I T H ( X - C H L O R O A C E T Y L -C H L O R I D E The syntheses of the compounds from groups A-,B and C. i l lus t ra -ted'below.startedwibh the appropriate amines R ^ - N H ^ and o C - c h l o r o -acetylchloride. O O " i ce -sal t bath ^ R 2 - N H 2 +• C 1 - C - C H 2 - C 1 ,1 J . >• R 2 - N H - C - C H 9 - C 1 + ( C ^ H g ^ N v H C l A 1 R 2 = a c)-methylphenyl b m-methylphenyl c p-methylphenyl d p-bromophenyl A 2 R ,^ •= a cyclohexyl b phenyl 8 B'2. = diphenylmethyl C 1 R 2 = 3, 5-dimethylphenyl A l l reactants were in equivalent mole quantities with a small excess of dry triethylamine to neutralize the HC1 evolved during the reaction. Table I OC - C H L O R O A C E T A N I L I D E D E R I V A T I V E S O II R 2 - H N - C - C H 2 - O l R 2 m . p . (-' C ) Y i e l d (%) o-methylphenyl 110 93 m-methylphenyl 92 89 p-methylphenyl 160 91 p-bromophenyl 182 90 cyclohexyl 105 90 phenyl 135-136 95 diphenylmethyl 128-129 • 92 3, 5-dimethylphenyl 140 91.5 The most important conditions for acylation in this reaction are perfectly dry reaction conditions and control of the reaction temperature. The reactions were, therefore, ca r r ied out in a flame-dried reaction flask at a temperature below 0 ° C . Reagent grade OC -chloroacetylchloride and triethylamine were commercia l ly available, but redistillation was necessary to obtain sufficiently dry starting mater ia ls . D r y non-polar solvents like ether and tetrahydrofuran could be used for the reaction since they do not solidify below 0 ° C as benzene does. However, dry ether was used exclusively because its low boiling point made it easy to remove the solvent. The -chloroacetamide derivative of each p r i m a r y amine was satisfactorily purified under reduced pressure in a sublimation apparatus. The compounds (Table i ) were stable at or above their melting points. The same synthetic route was followed using two other p r i m a r y amines, diphenylamine and 2, 6-dimethylaniline in order to begin to prepare C3 and D3 in F i g . 2 . In both cases, the same reaction temperature ( 0 ° /-^ 1 0 ° C ) was employed but the products were not stable. Therefore , the reaction temperature was dropped by re -placing the ice-sal t bath with a dry ice-acetone bath. Even at this temperature, the amides were slightly unstable. It was impossible to isolate the amides?;for the next reaction due to their instability. O II N H +• C I - C - C H 2 - C I > > tarry, dark brown product (i) ice-sal t bath, or (ii ) dry ice-acetone bath 0 II C H 3 f C 1 - C - C H 2 - C 1 • > tarry, blue-black product (i) i ce-sal t bath, or (ii) dry ice-acetone bath The synthesis of the compounds in group BI and 2 started respectively, with the reaction of unsymmetrical N,,N-dimethylethylene-10 diamine and benzoyl chloride, and N , N-dimethylethylenediamine and cyclohexanecarboxyl chloride: O O 2 C H , - N - C H ? - C H ? - N H ? + R - C - C l 1 C f " S x a l ^ b a t h ^ C H - N - C H - C H - N H - C - R 3 | 2 2 2 (Et ) z O 7 2 | 2 2 C H 3 C H 3 + C H . - N - C H . - C H - N H - H C 1 C H C H 3 R = phenyl or cyclohexyl The amides formed were separated, predried in air , and finally dried completely in an electrical oven ( 2 5 ° C / 1 0 m m . Hg). P A R T II N U C L E O P H I L I C S U B S T I T U T I O N W I T H D I M E T H Y L A M I N E The dimethylamino group was substituted for the J^-chlorine of the prepared ^[-chloroacetamides by nucleophilic substitu-tion to give the - d i m e t h y l a m i n o - N ' - R -acetamide derivatives: O N H - C - C H ? - C H ? - C 1 + 2 N H - C H „ . l c e " S a I t b a t h > N H - C - C H . - N - C H I L 2 | 3 " (Et) O and 2 f 3 R 7  C H - 2 _ R ? C H 2 3 T H F 2 3 + C H - N H - H C 1 C H 3 The boiling point of dimethylamine is 7 ° C . Therefore , the dimethyl-11 amine gas used was trapped in a graduated cylinder by a dry ice-acetone bath. Two equivalents of dimethylamine were used in each reaction. One equivalent was used for nucleophilic substitution, and the other one acted as a neutralizing agent for the hydrogen chloride. The reactions were conducted in an ice-sal t bath for 6 hours, and overnight at room temperature. A n acetone-dry ice condenser to prevent the escape of dimethylamine gas was not needed under the conditions used. The most common difficulty encountered with these reactions was the low solubility of the amides in dry ether. D r y tetra-hydrofuran (THF) or a mixture of T H F and dry methanol was employed as the solvent. The solvent was heated when this was necessary. Table II - D I M E T H Y L A M I N O - N - R 2 - A C E T A M I D E D E R I V A T I V E S O II R 2 - N H - - C - C H 2 - N - C H _ 1 C H 3 R 2 m . p . ( ° C ) b . p . , ° C ( m m . Hg) Y i e l d (%) o-methylphenyl . 57 - 58 .. - •• r 81 m-methylphenyl 109 (0.35) 95. 5 p-me thy Ipheny 1 103 (0.10) 90 p-bromophenyl 57 - 58 82 cyclohexyl 61 96 phenyl 135 (2.0) 85 diphenylme thy 1 89 3, 5-dimethylphenyl 83 125-127 (0.4) 91.5 R E D U C T I O N O F A M I D E S T O A M I N E S : lithium aluminum hydride (LAH) After obtaining the ^ -dimethylamino-N-R^-acetamide derivatives, they were reduced with lithium aluminum hydride to the corresponding ethylenediamines: L A H N H - C - N - C H 3 reflux ( E t ) 2 Q > N H - C H 2 - C H 2 - N - C H 3 R 2 C H 3 R 2 C H 3 Table III N , N - D I M E T H Y L - N - R 2 - E T H Y L E N E D I A M I N E N - C H . - C H , - N - C H , I 2 1 3 R 2 C H 3 R 2 b. p. 2 ° C ( m m . Hg. ) Y i e l d (%) o-methylphenyl 64-65 (0.1) 94 m-methylphenyl 70.0 (0.025) 95. 0 p-methylphenyl 65-68 (0.025) 95. 5 p-bromophenyl 90.0 (0.05) 95. 0 cyclohexyl 73.0 (2.0) 96. 5 phenyl 85.0 (1.0) 95 dime thy Ipheny 1 115.0 (0.25) 95 3, 5-dimethylphenyl 70. 0 (0.05) 94 To obtain the N , N-dimethyl -N' -cyclohexyl -methylethylene-diamine, N - c y c l o h e x y l - N - ^ 7 - N , N-dimethylaminoethyl)-cyclohexane-carboxamide (intermediate on the way to antergan analogue D - 4 in F i g . 2 ) was reduced with L A H to the respective amine: 13 N - Z / 6 - d i m e thy lphehyl ,/cyclohexylcarboxamide, the first compound in the synthesis of D-3 in F i g . 2 , by reacting cyclohexane-carboxyl( ichloride and 2, 6-dimethylaniline. It was attempted to reduce this compound using L A H in ether with refluxing, but this reaction was not successful even after 48 hours of refluxing. This was because of steric hindrance by the two methyl groups. When the solvent was changed to T H F and the reaction time was extended to six days, the amide was reduced. 14 L I T H I U M A L U M I N U M H Y D R I D E Although there are several ways to reduce amides, lithium aluminum hydride was employed exclusively in this work. It is necessary to keep in mind some characteristics of lithium aluminum hydride (referred to hereafter as L A H ) , and the unique way in which it and its derivatives reduce amides and nitriles to amines. L A H was discovered in 1947 by Finhalt , Bond and Schlesinger (i5<v). It was prepared as follows: 4 L i H f A 1 C 1 3 > L i A l H 4 +• 3 L i C l The reduction of compounds in ether solution is due to the transfer of a hydride ion f rom the L A H to the substrate. Ultimately, al l four hydrogen atoms are available as hydride. . ( A 1 H 4 ) " H ~ +• A 1 H 3 H +- I -L i H +• A 1 H > L i H - A l — H 6 I H These anions are nucleophilic reagents that normally attack polarized multiple bonds (e.g. C=0, C=N, N=0) at the positive atom; but isolated carbon-carbon multiple bonds in compounds like the 3-cyclohexene derivatives are not usually reduced. Generally, sodium borohydride is ineffective for the reduction of amides but it is a good reagent for the selective reduction of aldehydes, ketones and acid halides. The 15 reduction of amides with lithium aluminum hydride in ether produces amines in varying yields depending upon the structure of the amide (J57). O R - C - N - C H _ L 1 A I H 4 , excess ^ R - C H - N - C H . I 5 (Et) 0, reflux 2 | 3 CH„ C H , 3 A 3 R - C H - N - C H , R - C H = N - C H , b I > ' H 3 A I © * - C H 3 C H 3 (a) (b) Figure 3 • Mechanism of amide reduction The reduction mechanism for conversion of substituted amides to amines is believed to proceed by an initial reduction to a general aminoalcohol derivative (a in F i g . 3 ), followed by elimination and subsequent reduction of the iminium salt (or amine from a mono-substituted amide). If the reaction sequence could be stopped at the amino alcohol stage, this intermediate would be hydrolyzed to form an aldehyde (AHJ). The use of l imited amounts of l i thimum aluminum hydride and low temperatures brings about production of aldehyde, and better yield of the aldehyde is obtained by use of the less active reducing agent, L i A l H ( O C 0 H c ) n or L i A l H ( O C 0 H c ) „ at 0 ° C . Z Z b Z £ D 3 The use of refluxing and excess (1. 5-2 equivalents) of L A H is important in the reduction of n i t r i l e s t o the corresponding amines: •C = N 1. 5 equiv. L A H / / reflux, ( E t ) 2 Q . / ^ C H 2 N H 2 The reduction of 3-cyclohexenenitrile (above) was car r ied out using excess L A H (1. 5 equivalents) with refluxing. Microanalysis of the hydrochloride of the product confirmed its formula to be C ^ H ^ N C l . Spectral data gathered for the hydrochloride were consistent with 3-cyclohexenylmethylamine. However, when the 0(, ^ -unsaturated amide (a in F i g . 2 ) was reduced following the same procedure, both the amide and ^ , ^ —olefin were reduced (b). Therefore , L A H was not a suitable reducing agent for the amide with an , ^ _ bond. unsaturated C H 2 - C H 2 - N - C H 3 C H , L A H (Et) O, reflux (a) C H 2 - N - C H 2 - C H 2 - N - C H 3 C H , (b) Figure 4 Reduction of (X . — unsaturated amide with. L A H 17 Because of their relative stability and availability, L A H and N a B H ^ have been used as reducing agents in organic syntheses much more frequently than other complex metal hydrides have been. The many applications and limitations of L A H and N a B H ^ have been the subject of a number of reviewst'.J3.R,W0.)tLewis acids like aluminum chloride and boron trifluoride have been added to the complex metal hydrides to enhance the versatility of the reagents. These mixtures result in 'mixed hydrides,l('/QfcytfM). r.y^They••> have been prepared from various ratios of L i A l H ^ and A l C l ^ but those prepared with a hydride: halide ratio of 3:1, 1:1, 1:3 or 1:4 have been most frequently util ized in reduction reactions, as shown in the following equations: 3 L i A l H 4 f A 1 C 1 3 > 3 L i C l f 4 A 1 H 3 - (1) L i A l H 4 f A 1 C 1 3 > L i C l + 2 A1H4C1 - (2) L i A l H 4 f 3 A 1 C 1 3 > L i C l +• 4 A 1 H C 1 2 - (3) Unfortunately, rapid polymerization of the aluminum hydride formed in reaction (1) above ensues, and within minutes of mixing the reagents, the polymer begins to precipitate f rom the solution: n A 1 H 3 > ( A 1 H 3 ^ n The most common reducing agents include, in order of decreasing activity: lithium aluminum hydride, l ithium borohydride and sodium borohydride. Recently, sodium bis-(2-methoxyethoxy)-18 aluminum hydride (NaAlH^) (OCH^CH^-O-CH^)^ was introduced into the commerc ia l market under the names of R e d - A l (Aldrich Co. ) and Vitr ide (Eastman Kodak). The versatility, ease of use, stability, solubility, reactivity and selectivity of this reagent are claimed to be good m$jr463/4!7) *Bithium borohydride and especially lithium aluminum hydride react rapidly with hydroxyl compounds and, consequently, these metal hydrides must be used with purified, non-hydroxylic s o l -vents. Anhydrous ether and T H F are treated with sodium wire, or refluxed with lithium aluminum hydride and distilled through a Vigreux column. It is essential that the solvent be free of peroxides as wel l . A number of explosions have been reported with lithium aluminum hydride {/5Z ) and diethylether (/S3). Explosions with T H F are attributed to the presence of peroxides; those with diethylether are blamed on the presence of carbon dioxide. L A H should be weighed in a dry box and quickly transferred to the reaction vessel with a m i n i m u m of exposure to the atmosphere because a protective coating of aluminum hydroxide forms in humid air {/S^-). . If the amides are easily soluble in ether and T H F , solutions are added dropwise through a dropping funnel into the L A H solution. A Soxhlet extractor is used to maintain dissolution of sparingly soluble amides, which are difficult to add to the reaction flask using a dropping funnel. Excess lithium aluminum hydride remaining in the reaction 19 mixture is decomposed by dropwise addition of ethanol, water-saturated ether, or distil led water. Although addition of ethyl acetate is some-times employed, this procedure is not advisable when the product is an amine since the amine itself may react with ethyl acetate {/&]). It is customary to add to the reaction mixture the calculated amounts of water or dilute aqueous sodium hydroxide required to convert lithium and aluminum salts to granular lithium aluminate ( L i A l O - , ) . During the decomposition of excess L A H , it is important not to add excess water since its presence wil l convert the reaction mixture to an emulsion of the solvent, water and aluminum hydroxide which is difficult to filter or extract. A n alternative isolation procedure employs an aqueous solution of sodium potassium tartrate to hydrolyse and complex the aluminum salts present in the reaction mixture. In the present work, excess L A H was usually hydrolyzed by a small excess of distil led water and 40% sodium hydroxide was added for clear separation. The solution was separated by centrifugation or decantation. The amide was reduced to the amine and lithium aluminate was produced as a by-product. Li thium aluminate and excess L A H were decomposed by the addition of distil led water: / . 2 R C O N H 2 +• L i A l H 4 > 2 R C H 2 N H 2 4- L i A l C > 2 2 L i A 1 0 2 + H 2 0 > 2 L i O H 4- A 1 2 0 3 4- H 2 L i A l H 4 4- 4H 2 0(excess) > L i O H 4- A l ( O H ) 3 4- H 2 L i A l H 4 ( e x c e s s ) + 2 H 2 Q »• L i A 1 0 2 +- 4 H 2 f 20 F O R M A T I O N O F A M I D E S : (Table IV) The secondary amines (Table III ) were reacted with cyclohexane carboxyl chloride in the presence of dry triethylamine, The products of these reactions were the final intermediates of antergan analogues A - l a, b, c, d, e and C - l in F i g . 2 . O II C + H N - C H 2 - C H 2 - N - C H 3 -i ce - salt bath C l I R 2 CH, (E t ) 3 N O II C - N - C H , - C H 7 - N - C H , I L c I J R 2 C H , + ( E t ) 3 N - H C l Table IV N - R 2 - N - ( / # - N , N - D I M E T H Y L A M I N O E T H Y L ) -CY C L O H E X A N E C A R B O X A M I D E O II C - N - C H _ - C H , - N - C H _ I 2 2 | 3 R , C H , R 2 m . p. ( ° C ) b . p. ° C ( m m . Hg) Y i e l d (%) o-methylphenyl 125-125 (0.15) 92.4 m-methylphenyl 116-118 (0.05) 85. 0 p-methylphenyl 128-130 (0. 125) 89 p-bromophenyl 81-82 80 cyclohexyl 165 (5.0) 74 3, 5-dimethylphenyl 145 (0.4) 81 21 To obtain the final intermediate for antergan analogue C -2 , N , N-dimethyl-N'-phenylethylenediamine was reacted with 1-cyclohexenecarboxyl chloride: CV - C - C 1 + H N -C H . - C H , - N - C H , f T > / ^V-C-N-CH - C H - N - C H n CH. ^ j j C H 3 I II C H3 + (Et) N ' . H C l S i m i l a r l y , 3-cyclohexenecarboxyl chloride was reacted with N , N-dimethyl -N' -R^-e thylenediamine in the presence of triethylamine. The products of these reactions were the final intermediates of antergan analogues A - 2 a and b. O O f, \ II i ce-sal t bath \ II (/ V - C - C l + N H - C H 2 - C H 2 - N - C H 3 ( E t ) 3 N *\ >C-N-CH 2-CH 2-N-CH 3 R 2 C H 3 R 2 C H 3 + ( E t ) 3 N « H C l 22 Table V N - R ^ - N - ( - N , N - D I M E T H Y L A M I N O E T H Y L ) -3- C Y C L O H E X E N E C A R B O X A M I D E R 2 b . p . ° C ( m m . Hg) Y i e l d (%) Phenyl 120 (0.3) 82 Cyclohexyl 160 (0.65-0.75) 96 N , N-dimethyl-N'-benzylethylenediamine was reacted benzoyl chloride in the presence of triethylamine to f o r m the final inter-mediate of the antergan analogue B - l in F i g . 2 : < ^ ~ ^ - C H 2 - N H - C H 2 - C H 2 - N - C H 3 C H , O ice-sal t bath (E t ) 3 N C H 2 - N H - C H 2 - C H 2 - N - C H 3 c = o C H , ••fc ( E t ) 3 N - H C l N , N-dimethyl-N'-cyclohexylethylediamine was reacted with benzoyl chloride in the presence of triethylamine to form the final intermediate of antergan analogue A - 3 . 23 P R E P A R A T I O N O F A L I C Y C L I C A C I D C H L O R I D E S In order to make cyclohexanecarboxyl and 3 -cyclohexenecar-boxyl amides, commercial ly available 3-cyclohexenecarboxylic acid and cyclohexanecarboxylic acid were converted into their acid chlorides by the procedure of Cope and Ciganek {!£&). 1 - cyclohexenecarboxylic acid was prepared in this work, and also converted into the acid chloride to make other amides. The substitution of chlorine for the hydroxy of the above acids with" tbionyf chloride was an endothermic reaction. Other methods of formation of acid chlorides are reaction of carboxylic acids with phorphorous pentachloride or phosphorous tr ichloride. R - C O - O H + P C 1 5 —> R - C O - C 1 f H C l + P O C l 3 3 R - C O - O H + P C 1 3 > 3 R - C O - C l 4- H3PO3 R - C O - O H +' S O C l ^ — * R - C O - C 1 +- S O z f H C l The choice of reagent for preparation of an acid chloride was largely determined by the relative boiling points of the acid chloride and the 24 by-products for the convenience of separation of the products by fractional distillation. R E D U C T I O N O F T E R T I A R Y A M I D E S The N - R 2 ~ N - - N , N-dime thylaminoethyl)- cyclohexane-carboxamide were reduced by L A H with refluxing to yield analogues of antergan A - 1 a, b, c, d and e: / V C - N - C H - C H , - N - C H , ^ A H > / ) - C H - N - C H . - C H - N - C H , \ J I 2 2 j 3 (Et ) 2 0 , reflux \ / 2 , 2 2 | 3 R 2 C H 3 R 2 C H 3 R 0 as in Table VI . Table VI N , N - D I M E T H Y L - N ' - C Y C L O H E X Y L M E T H Y L - N ' - R z -E T HY L E N E D I A M I N E S ( A N A L O G U E S O F A N T E R G A N ) R 2 b . p . ° C ( m m . Hg) Y i e l d (%) 0 - m e thy Ipheny 1 126-128 (0.4) 97 m-methylphenyl 140 (0.5) 96 p-methylphenyl 138 (0.8) 97 p-bromophenyl 150 (0.2) 37. 34 cyclohexyl 73 (2.0) 95 When the N - p - b r o m o p h e n y l - N - ( ^ - N , N-dimethylamino-ethyl)-car clyhexanecarboxamide was reduced by an excess of L A H with refluxing for 12 hours, the corresponding tertiary amine ( N , N -d i m e t h y l - N ' - cyclohexylmethyl-N'-p-bromophenylethylenediamine) was formed. The yield was only 37.4%; this was because cleavage of the C - N bond resulted in the formation of a secondary amine and cyclohexylmethylalcohol as by-products : O ( ^ } - C - N - C H 2 - C H 2 - N - C H 3 ( E ^ ^ e f l ^ C H , - N - C H 9 - C H , - N - C H „ r fl C H 3 B r O C H 2 - O H C H 3 + N H - C H _ - C H _ - N 2 c \ C H 3 In order to obtain the analogues of antergan A - 2 a and b in ( F i g . 2 , N - R 2 - N - ( ^ - N , N-dimethylaminoethyl)-3-cyclohexenecarbox-amides were reduced with L A H : O ^ ^ - C - N - C H 2 - C H 2 - N - C H 3 R 2 C H 3 L A H ^ < f ~ \ r H - N H - C H , - C H 7 - N - C H , (Et ) 2 0, reflux \ / 2 ^ L C H 26 Table VII N, N - D I M E T H Y L - N ' - 3 - CY C L O H E X E N Y L -M E T HY L E T HY L E N E D I A M I N E ( A N A L O G U E S O F A N T E R G A N A - 2 a A N D b) R 2 b . p . 0 ° C ( m m . Hg) Y i e l d (%) Phenyl 162-164 (2.0) 94 Cyclohexyl 116 (0.6) 96 In order to obtain the analogues of antergan B - l and 2 in F i g . 2 , N-benzyl-N-(^-/^{(-dimethylaminoethyU-phenylcarboxamide and o( - N , N-dimethylamino-N-diphenylmethylacetamide were reduced with L A H : ^ ~ ^ - C H 2 - N - C H 2 - C H 2 - N - C H 3 C=0 C H , L A H /T " "^ reflux, (Et) O \ — / I 2 I 5 C H 2 C H 3 C H - N H - C H - C H - N - C H , " r e i m x , \ i \ i ) 7 ^ > \ — / i r ^ T i C H . O < * f l C H 3 reflux, ( E t l ^ X ^ J T " " 2 — 2 " 7 ™ 3 I1 J. J. I In order to obtain the analogue of antergan A-3 , .N ' -cyc lohexyl -N-'C^'- 'Ni N-dimetTiylaminoethyr)-benzene.carboxamide were reduced with L A H : o ^ 7 - C - N - C H ? - C H ? - N - C H o ,^ t L A H r ) ^ \ c H - N - C H - C H , - N - C H , X=J > k I.. 3 (Et ) z O, r e f l u x 2 ^ 2 2 ^ 3 c h 3 r ^ i P R E P A R A T I O N O F l - C Y C L O H E X E N E C A R B O X Y L I C A C I D The synthesis of 1-cyclohexenecarboxylic acid began with the preparation of cyclohexyl cyanohydrin by a method similar to that reported in ref. /6? : Q = 0 • K C N _ , Q( O H + K C l +- cone. HC1 C N The reaction was conducted in the fume hood, because of the possibility of hydrogen cyanide production. Although this very toxic gas smells like almond o i l , it soon de-sensitizes the sense of smel l . The cyclohexyl cyanohydrin was esterified at room temperature ( 2 5 ° C ) with dry methanol which had been saturated with hydrogen chloride in an ice-salt bath: , , O H , O H C=N C=0 I O - C H 3 + N H 4 C 1 The methyl 1-hydroxycyclohexancarboxylate was dehydrated with thionyl chloride in the presence of pyridine: 28 The methyl 1 -cyclohexene carboxylate was hydrolyzed by 10% sodium hydroxide solution, and neutralized with dilute hydrochloric acid to yield free cyclohexenylcarboxylic acid: o o o / ^ 11 reflux / 11 d i l . H C l v / ^ H < V - C - O C H , f • ( V~ C - O N a „ r f ^ V ( xV - C - O H \ / 3 ! 0 % N a O H \ / 2 5 ° C \ / A T T E M P T T O P R E P A R E A M I D E S USING A C A R B O D I I M I D E An adaptation of the carbodiimide synthesis of Sheehan {/S3 ) and Vigneau {I6H-), in which a carboxylic acid reacts with an amine to produce a carboxamide and N , N' -dicyclohexylurea , was tried in an effort to find a simple and convenient method for amide formation. In one t r ia l , one equivalent of each of N , N-dimethyl -N' -cyc lohexyle thyl -enediamine and cyclohexanecarboxylic acid were dissolved in methylene chloride. A solution of one equivalent of dicyclohexylcarbodiimide was added dropwise into the reaction mixture over a period of five minutes with rapid s t i r r ing . At the end of the addition, the reaction mixture became cloudy. This procedure was car r ied out in different runs at room temperature, and at 0 ° C ; the reaction was continued for two hours. = C N H H + O' / c O D N H = " c " - N H - / \ N H - C H * - C H * - N B O N = C - N H r l O H c = o <* o r NH* - C H z - C H 2 - M - C H 9 1 C H 3 o o II N - C - N H I c = o N H - C - N H - ^ ^ and t O O < U - c - Q Figure 5 . Mechanisms for reaction between carboxylic acid and carbodiimide 30 In a second tr ial , the solution of carboxylic acid was added dropwise to a mixture of diimide and N , N-dimethyl-N'-cyclohexylethylenediamine. In these reactions, the amide was not formed. Instead, dicyclohexanecarboxylic^anhydride was the main product and dicyclo-hexylurea was a by-product . The ir N - H bending band at 1625 cm * of dicyclohexylurea (m.p . 2 2 3 ° C ) was used to identify its presence. Disappearance of the stretching band characteristic of -N=C=N- at 2115 cm ^ was taken as a proof that the dicyclohexylcarbodiimide was used up. Separation of the anhydride f rom the dicyclohexylurea was not attempted. Dicyclohexylurea is sparingly soluble in the solvents used in the reactions: dioxane, T H F , benzene, methylene chloride, ether and hexane; but the urea was dissolved in sufficient quantities to be a contaminant of the acid anhydride. In F i g . 5 , the cation in A , and anions of the acids are the reaction entities, and the relatively weak nucleophilic secondary amine (N, N-dimethyl-N' -cyclohexylethylene-diamine) is probably less active than the cyclohexanecarboxylic acid anion during the attack on the cationic carbon atom in D . The amide formation f rom cyclohexanecarboxylic acid and N , N - d i m e t h y l - N ' - c y c l o -hexylethylenediamine was attempted using various solvents (dioxane, T H F , hexane, and benzene) after methylene chloride was unsuccessful . A l l these reactions were car r ied out at two different temperatures, 0 ° C and room temperature; but all experiments failed to pro duce fi - N , N -d i m e t h y l e t h y l - N ' - c y c l o h e x y l - N 1 - cyclohexanecarboxamide. 31 The reaction between carboxylic acids and carbodiimides follows the pattern in F i g . 5 . The first step is the attack of a proton to give the cation A . This is followed by the attachment of the acid anion, which results in the formation of B . This can rearrange to f o r m an acylurea C . Alternatively, attachment of a second proton can proceed more quickly than the oxygen-to-nitrogen migration and the cation D is produced; D is subsequently attacked by the acid anion. The products of the reaction generally depend on the nature of the carbodiimide, the acid, solvent, and the temperature at which the reaction is car r ied out. P R E P A R A T I O N O O F C Y C L O H E X A D I E N E S Y S T E M In order to replace the benzyl group of antergan by cyclo-hexadiene, preparation of the appropriate hexadienecarboxylic acid was attempted by reduction of the aromatic ring of benzoic acid. B i r c h reduction and the D i e l s - A l d e r synthesis were employed for the preparation of 2, 5-cyclohexadiene-1 - carboxylic acid, and 3-chloro-4-cyclohexene-1, 2-dicarboxylic acid anhydride, starting materials respectively for antergan analogues N , N - d i m e t h y l - N ' - 2 , 5-cyclohexa-dienylmethyl-N'-cyclohexylethylenediamine, and N , N - d i m e t h y l - N 1 -1, 4-cyclohexadienylmethyl-N'-cyclohexylethylenediamine. P R E P A R A T I O N O F 2, 5 - C Y C L O H E X A D I E N E - 1 - C A R B O X Y L I C A C I D The most important use of metal -ammonia reducing agents was discovered by Wooster (139). B i r c h made a thorough study of the reduction of benzene derivatives and developed practical applications for the reaction which now bears his name. The reduction mechanism was proposed by him (/S9): Figure 6 . Mechanisms of B i r c h reduction of benzoic acid and aniline. In the metal ammonia reducing solution, benzene derivatives are equilibrium with a low concentration of the radical anion (a) in F i g . 6 . A suitable acid (ammonia is too weak) is able to displace the equil ibrium towards the formation of another radical (b). A second electron is added to the second radical to give another anion (c), which is protonated i r r e v e r s i b l y under the reaction conditions to give cyclohexa-2, 5-diene as the major product. The electron density would be greatest at positions and C4 to the electron-attracting sub-stituent, and preferential protonation of the radical-anion at these positions is to be expected. S imilar ly , an electron-releasing sub-stituent would cause the radical anion to be protonated in the and positions. In a benzene derivative like benzoic acid which has an electron-withdrawing group, the benzene ring is reduced in the and C4 positions because of the substituent. The reduction of benzoic acid by sodium-ammonia was carr ied out by the following method i l i q . N H 3 ( _ 3 3 ° C ) O H Na, E t O H H H H O O H The product, 2, 5-cyclohexadiene-1 - carboxylic acid was purified by distillation through a Vigreux column, or alternatively through a spinning column which gave more efficient separation. The n m r spectrum showed that one proton at and two at were coupled, comprising a multiplet with an unusually large coupling constant/#f).The product obtained in every run contained traces of benzoic acid in the n m r spectrum. It was thought at f irst that imperfect dryness of the 34 ethanol was responsible for the presence of benzoic acid in the product. Therefore , the following methods were used to obtain completely dry ethanol. Diethylphthalate reacts i r r e v e r s i b l y with sodium ethoxide and water ( / S i ): C O O C 2 H 5 a reflux +• C 2 - H 5 - O N a + H 2 0 > R C O O N a + 2 C 2 H 5 O H C O O C 2 H 5 Alternatively, water can be removed by reaction with magnesium ethoxide: M g ( O C 2 H 5 ) 2 +- 2 H z O >Mg (OH) 2 + 2 C 2 H 5 O H Since the magnesium hydroxide is insoluble in ethanol, the reaction proceeds to completion (162-); but even when the ethanol was dried using these methods, some benzoic acid remained in the product of the reduction. It was believed that improper distillation methods might be responsible. However, after many runs using good distillation con-ditions, it was concluded that spontaneous aromatization of 2, 5 -cyclo-hexadiene-1 -carboxylic acid was probably occurring during the distillation. N m r and i r spectra showed that the product obtained was sufficiently pure. The 2, 5 cyclohexadiene-1-carboxylic acid was 35 converted to its acid chloride, and reacted with cyclohexylamine to prepare N-cyclohexyl-2 , 5 cyclohexadiene-1-carboxamide. The n m r and i r spectra of the amide were satisfactory, but the result of microelemental analysis was different f rom its calculated value. • H C l Subsequently, reduction of N-cyclohexyl 2, 5-cyclohexadiene-1 - car-boxamide was carr ied out using L A H with reflux. The c a r b o x y l . group was reduced, but the 2, 5-cyclohexadiene was oxidized to an aromatic r ing . Therefore , this method was not satisfactory for the synthesis of the antergan analogue it was designed for (N, N - d i m e t h y l - N ' - 2 , 5 -cyclo-hexadienylmethyl-N 1 - cyclohexylethylenediamine). P R E P A R A T I O N O F 3 - C H L O R O - 4 - H E X E N E - 1, 2 - D I C A R B O X Y L I C A C I D A N H Y D R I D E In the first step of the synthesis of 1, 4-cyclohexadiene carboxylic acid, 1-chlorobutadiene and cis-butendioic acid anhydride (maleic anhydride) were reacted in the presence of iodine and hydro-quinone to produce 3-chloro-4-cyclohexene-1 , 2-dicarboxylic acid anhydride. c l ci (I) c i s - 1 - chlorobutadiene (S-c is - isomer) Ci (II) cisoid conformation Ci ± f (III) trans-1-chlorobutadiene (S-trans - isomer) (IV) cisoid conformation Figure 7 . Geometrical isomers of 1-chlorobutadiene The S-c is - isomer (I) does not convert to the cisoid conformation (II) because of steric hindrance. F o r the same reason, it does not enter the diene synthesis ( D i e l s - A l d e r r e a c t i o n ) (fll ). The S - c i s - i s o m e r (I) converts easily to the S-trans - i s o m e r in the presence of iodine, which yields a cisoid conformation. The mixture of S - c i s - and t rans- isomer s of 1 - chlorobutadiene can, therefore, be utilized to react with maleic anhydride in the presence of iodine. Diene synthesis was found to occur readily between the cisoid conformation (IV) and maleic anhydride: + \ C / The synthesis of 3-chloro-4-cyclohexene-1, 2-dicarboxylic acid anhydride was confirmed by i r, n m r, and elemental analyses. The preparation of 1, 4-cyclohexadiencarboxylic acid f rom the anhydride was not attempted, although this reaction is possible (170 ). Nothing further was done with the anhydride because it was believed the synthesis of N , N - d i m e t h y l - N 1 - c y c l o h e x y l - N 1 - 1, 4- cyclohexadienylmethylethylenediamine would be impossible, after the reduction of N-cyclohexyl -2 , 5-cyclohexa-dienecarboxamide to its amine was found to be impossible (p. 3$) 38 H A L O D E C A R B O X Y L A T I O N O F A C A R B O X Y L I C A C I D The halodecarboxylation of a cyclohexanecarboxylic acid using lead tetraacetate was car r ied out, following the method of Kochi (168): // V . C / / 0 ¥ P b ( A c O ) 4 +• L i C l p N Z -> V > C l+Pb ( O A c ) ? +• A c O H ^ Benzene \ / & 4 - L i O A c + C O z t The reaction was conducted by adding one equivalent of halide to a solution of acid and lead acetate in benzene. The mixture was refluxed with st irr ing under nitrogen gas. Although the reactants were initially heterogeneous, the reaction was completed within five minutes. Moisture in the air inhibits the reaction and flushing with nitrogen was necessary for a good y ie ld . SYNTHESIS O F - C H L O R O - N , N - D I M E T H Y L A C E T A M I D E The preparation of antergan analogues using chloroacetyl-chloride gave good yields, but requires many further reactions to get to the analogues. In an effort to find a simple and efficient method for synthesis of the N-dimethylethylene part of antergan analogues, ^ -c h l o r o - N , N-dimethylacetamide was synthesized: 39 n < E t > 2 ° ° 2 N H - C H 3 + C 1 - C - C H 2 - C 1 d r y i c e-acetone bath' C H 3 - N H - C - C H 2 - C l C H 3 C H 3 C H I 5 ' • f C H 3 - N H - H C 1 A s the reaction is very vigorous, it was conducted carefully at a low temperature ( - 8 0 ° C , in dry ice-acetone bath) using stoichiometric equivalents of the reactants. The reaction of =K - c h l o r o - N , N-dimethyl-acetamide with benzylcyclohexylamine, however, which would have provided an intermediate for the formation of the N-dimethylamine part of the analogues, gave a poor y ie ld . This method, therefore, does not appear to have any advantages over the methods using chloroacetylchloride reaction. In order to prepare antergan anlogue C-2b (Fig . £ ), the direct condensations indicated below were attempted. When it was found that the reaction forming (a), the final intermediate in the for-mation of analogue C-2b would not go ahead, a condensation using the p r i m a r y amine was attempted. Compound (b), which would also have been an intermediate in the synthesis of analogue C-2b, was not formed either. Excess amine (1.2 times the equivalent of the ester present) was added dropwise to the ester solution. Even after refluxing for 3 days, no new compound could be detected on a thin layer silicate plate. 40 A T T E M P T T O A L K Y L A T E A S E C O N D A R Y A M I N E WITH C Y C L O H E X Y L p - T O L U E N E S U L F O N A T E Cyclohexyl tosylate was formed in good yield in the following reaction: 41 A l k y l sulfonates are generally good alkylating agents; therefore, alkylation of the secondary amines N , N - d i m e t h y l - N 1 - c y c l o h e x y l m e t h y l -enediamine (a in diagram below) and N , N-dimethyl -N' -benzylethylene-diamine (b in diagram) with cyclohexyl tosylate was attempted. (a) (b) no product no product It appeared that a small amount of a quaternary salt was formed during these reactions, but it was not identified. The low reactivity of cyclohexyl tosylates has been attributed to steric hinderance by the cyclohexyl group_ Alkylation of N , N-dimethyl-N'-benzylethylenediamine with benzyl bromide and benzyl chloride was then attempted. These reactions were used to tell whether the secondary or tertiary amine site of the diamine is most active; since the quaternary ammonium salt was formed exclusively, it is concluded that the tertiary amine site is more active than the secondary site. H I C H C H - N - C H - - C H - N + C l - C H 2 5 ° C T H F C H , C l H I © I - C H 2 - N - C H 2 - C H 2 - N - C H 2 C H 3 W rCS In the same way, alkyl halid.es generally undergo nucleophilic substitution readily, while aryl halides are extremely slow in substitutions under the same reaction conditions. Cyclohexyl chloride and 1-chloro-3-cyclohexene, like cyclohexyl-p-toluene sul -fonate, were not active alkylating agents for N , N - d i m e t h y l - N 1 -cyclohexylethylenediamine, for the same reason: C l (' W c H - N - C H - C H . - N - C H , 2 2 2 | 3 C H no product C l no product In another t r ia l , the sodium salt of N , N-dime thy I - N 1 -cyclohexylethylenediamine was made by reaction with sodium amide, and 1-chloro-3-cyclohexene was added into the reaction mixture . The reaction was continued for 24 hours. The yield of tertiary amine was very poor. This may be attributed to the low activity and steric hindrance of 1 - chloro-3-cyclohexene. 44 N a N H 2 , N 2 Na C H 2 - N H - C H 2 - C H 2 - N - C H I C H 3 3 benzene(reflux) > POOT Yield. A T T E M P T E D C O N D E N S A T I O N O F P R I M A R Y A M I N E S W I T H fi - D I M E T H Y L A M I N O E T H Y L C H L O R I D E H Y D R O C H L O R I D E The synthesis of N , N-dime thy 1-N'-cyclohexylethylene-diamine, an intermediate for antergan analogue A - 3 in F i g . £ , was attempted by reacting one equivalent of cyclohexylamine with two equivalents of sodium amide. After the formation of the sodium salt of cyclohexylamine had been confirmed, one equivalent of ^ - dime-thylamino ethy 1c chloride* H C l was added. The yield of the desired product (N, N-dimethyl-N'-cyclohexylethylenediamine) was very poor, and the product consisted almost entirely of N , N-dimethylethylene-diamine. One reason for this is probably that the sodium salt of cyclohexylamine neutralized the hydrochloride salt first (reaction a. in figure below); returning the amine salt to cyclohexylamine, producing sodium chloride, and eliminating the possibility of reaction b. occurr ing to any significant extent. This leaves the extra equivalent of sodium amide free to react with ^ -dimethylaminoethyl chloride (reaction c) to give the N , N-dimethylethylenediamine. Evidently, the sodium salt of cyclohexylamine is a very strong base, perhaps even stronger than sodium amide. Another factor decreasing the yield of the desired 45 product is undoubtedly cyclization of the fl -dimethylaminoethyl chloride formed in reaction b, to give the quaternary ammonium salt (reaction d). Separation of the two products of this reaction (N, N -dimethylen'ediamine and N , N-dimethyl-N'-cyclohexylethylenediamine) was difficult on a 9 - inch Vigreux column. NH-4- 2 N a N H , benzene, N 2 reflux C H , N H 2 - C H 2 - C H 2 - N - C H 3 (c) C 1 - C H 0 - C NaNH 2 (ex t ra equivalent)' C H , H 0- h- C H , 4-C H , C H , •N' X C H / N C H 3 CH» I C£ -CHit- CHr N - C H 3 " N H 3 f NaC 1 NHNa N H - C H 2 - C H 2 - N - C H 3 C H , NHNa +- N a N H 2 4- N H 3 ' T N H Z C H , C 1 - C H 2 - C H 2 - N * HC1 4- N a C l ~ (a) ( b ) C H , N a N H , 46 P A R T III A N A L Y T I C A L M E T H O D S A Beckman I . R . - 1 0 Infrared Spectrophotometer (Beckman Instruments, Inc. ) was used to record the infrared spectra. L iquid samples were scanned as thin films between N a C l plates and solids were incorporated into K B r discs . Nuclear magnetic resonance (nmr) spectra were recorded using a V a r i a n T-60 Spectrometer. The solutions were approximately 10% and deuterated chloroform ( C D C l ^ ) was used as solvent. Tetramethylsilane (TMS) was used as an internal standard. Peaks in the nmr spectra are reported according to the following format: chemical shifts from T M S are expressed in dimen-sionless units & , peak multiplicity, number of protons, coupling constant where this is applicable, position of protons in the molecule. The following abbreviations are used for the peak multiplici ty : s, singlet; d, doublet; t, triplet; m , multiplet. Melting points were determined, unless otherwise indicated, in open capillaries in a T h o m a s -Hoover Unimelt apparatus (Arthur H . Thomas C o . ) . A l l melting points are reported uncorrected. 47 •PART IV E X P E R I M E N T A L A , SYNTHESIS O F N, N - D I M E T H Y L - N ' - C Y C L O H E X Y L M E T H Y L -N ' - o - M E T H Y L P H E N Y L E T H Y L E N E D I A M I N E amine (30. 34g, 0. 3 mole) in 500 m l . of dry ether was placed in a 1 litre three-necked flask. The flask was equipped with a mechanical s t i r rer , a side a r m for setting a thermometer (range -100 to 5 0 ° C ) and a dropping funnel (1Z5 m l . ) which car r ied a drying tube. The solution was cooled by st i rr ing for 1 hour in an ice -salt bath (-5 to - 1 0 ° C ) . A solution of chloroacetyl chloride (34 g, 0. 3 mole) in 100 m l . of dry ether was added from the dropping funnel to the reaction flask. The solution was added slowly, with constant s t i rr ing, so as to keep the temperature of the solution below 0 ° C . The mixture was s t i rred overnight at room temperature. Then 100 m l . of distil led water was added to the flask, and the mixture was st irred for 15 minutes. The white product was suction filtered through a Buchner funnel. The ethereal layer was then separated and flash evaporated to remove the solvent. The remaining solid was combined with the suction fil tered product, and placed in 500 m l . beaker. 5% HC1 (100 m l . ) and cold distil led water (100 m l . ) were added, and the product was then suction 1. A solution of o-methylaniline (32. l g , 0. 3 mole) and tr imethyl-filtered and washed with distil led water until no H C l could be detected using litmus paper. The product was predried in the air and finally dried at 4 0 ° C in an oven under reduced pressure (20 m m Hg). The product was purified by reduced pressure sublimation. Y i e l d : 51. 6g (93%). m . p . : 1 1 0 ° C . i r (KBr) 3260 (N-H), 3040 (C-H) , 1600 (C=0), 760, 730 c m " 1 ( C - C l ) ; nmr (CDC1 3)^.7. 33 - 7. 07 (m, 4, phenyl-H>, 4. 20 (s , 2.) ( C H 2 - C 1 ) , 2.27 (si, 3, to-CH3), 1. 53 (s, 1, N - H ) . A n a l . Calcd . for C g H ^ N C I O : C , 58.9; H , 5.45; N , 7.64; C l , 19.32. Found: C, 58.83; H , 5.56; N , 7.72; C l , 19.2. 2. o{ -Dimethylamino-N-o-methylphenylacetamide DryDether (350 m l . ) was placed in a 500 m l . three-necked flask equipped with a dropping funnel (125 m l . ) with drying tube, a mechanical s t i r rer and a side a r m for setting a thermometer (range -100 to 5 0 ° C ) . The solution was cooled to between -5 and - 1 0 ° C in an ice-sal t bath for 1 hour. Dimethylamine (26.5 cc . , 0.4 mole) was trapped in a dry graduated cylinder (50 m l . ) f rom a dimethylamine gas tank by an acetone-dry ice bath ( - 8 0 ° C ) , and poured al l at once into the three-necked reaction flask. A solution of o( - c h l o r o - o - m e t h y l -phenylacetamide (27. 55g, 0. 15 mole) in dry tetrahydrofuran (80 m l . ) was added slowly from the dropping funnel to the vigorously s t i rred solution so as to keep the temperature of the reaction mixture below 49 0 ° C . After the addition was completed, s t irr ing was continued. The temperature was kept below 0 ° C for 10 hours, and below 1 0 ° C overnight. The white dimethylamine salt was suction filtered and the solvent was removed using a rotary evaporator. 30 m l . of disti l led water was added to the product, and the mixture was shaken vigorously in the separating funnel. The mixture was extracted with two 150 m l . portions of ether, and the combined ethereal solution was dried over anhydrous magnesium sulfate. The solution was suction filtered and the solvent was removed using a rotary evaporator. The product was further disti l led through an a i r - cooled double-surface condenser. . The product was recrystal l ized from petroleum ether ( 3 0 ° - 6 0 ° C ) . Y i e l d : 23g (81%). m . p . : 5 7 - 5 8 ° C . i r (KBr) 3280 (N-H), 2970, 2940, 2820, 2770 ( N - ( C H 3 ) 2 ) , 1670 (C=0), 765, 720 c m " 1 (phenyl-H); nmr ( C D C 1 3 ) £ 8 . 5 - 8 . 1 3 (broad s;, 1, N - H ) , 7.33 - 6.93 (m, 4, phenyl-H), 3.07 (s', 2, 0 = C - C H 2 ) , 2.37(s., 6, N C C H ^ ) , 2.23 (Is. 3, O - C H j ) . A n a l . . Calcd . for C . . H , , N o 0 : C, 68. 72; H , 8. 39; 11 1 o 2 N , 14.57. Found: C, 68.92; H , 8. 34; N, 14.49. 3. N , N-dimethyl-N'-o-methylphenylethylenediamine A dry 250 m l . three-necked flask was equipped with a mechanical s t i r rer , dropping funnel (125 m l . ) and reflux condenser. The funnel and condenser both car r ied drying tubes. 150 m l . of dry ether and lithium aluminum hydride (7. 6g, 0. 2 mole) were added to the ether, and the mixture was gently refluxed with s t i r r ing for 3 hours A solution of ck -Dimethy 1 a m i n o - N - o -methylphenylacetamide (19. 2g, 0. 1 mole) in 50 m l . dry ether was placed in the dropping funnel and added dropwise at a rate that maintained gentle reflux. After the addition was complete, the mixture was refluxed for 40 hours with s t i r r ing . When the reduction was completed, the reaction mixture was cooled in an ice-sal t bath for 30 minutes, and 20 m l . of disti l led water was added dropwise to the rapidly s t i rred mixture in order to decompose the excess hydride. The st irr ing was continued until the mixture became white in colour. Then sufficient 40% N a O H solution was added to allow clear separation of the ethereal layer . The ether-insoluble residue was separated by centrifugation. The ethereal layer was dried over anhydrous magnesium sulfate overnight. The solvent was removed by flash evaporation and the residue was distil led under reduced pressure using a 9 inch Vigreux column and a heating tape on an oil bath. The fraction boiling at 6 4 ° - 6 5 ° ( 0 . 1 m m . Hg) was c o l -lected. Y i e l d : 16. 8g (94. 1%). i r (neat) 3380 (N-H), 2920, 2860, 2820, 2760 c m - 1 ( C - H ) ; nmr ( C D C ^ ) ^ 7.27-6.43 (m, 4,phenyl-H), 4. 47-40 (broad a, 1, N - H ) , 3.30-2.93 (m, 2, N H - C H 2 - C ) , 2.70-2. 33 (m, 2, - C - C H 2 -N C . C H 3 ) 2 ) , 2.23 ('s, 6, - N C C H 3 ) 2 ) , 2.10 (s, 3, 0 -CH3) . 51 Di-hydrochloride derivative Into a solution of the tertiary amine (2.91g, 0.02 mole) in 100 m l . dry ether, dry hydrogen gas was passed f rom a cylinder through a concentrated sulfuric acid. When salt formation was completed, the solid was suction filtered under a stream of dry nitrogen gas to prevent it from absorbing moisture from the a i r . The salt was washed with 100 m l . dry ether on the Buchner funnel. The hydrochloride was completely dried, and recrys ta l l ized f rom dry ethanol and ether to give colourless crystals ; m . p . 1 9 0 ° C . A n a l . C a l c d . for C j l H 2 Q N 2 C 1 2 : C , 52. 595; H , 8.03; N , 11.15; C l , 28.22. Found: C , 52. 50; H , 7.94; N , 11.09; C l , 28.30. 4. N - o - m e thy Ipheny l - N - ( - N , N-dimethylaminoethyl)- cyclohexanecarboxamide A dry 250 m l . three-necked flask was equipped with a mechanical s t i r rer , dropping funnel (125 m l . ) with drying tube, and a side a r m for setting a thermometer (range -100 to 5 0 ° C ) . . N , N -dimethyl-N'-o-methylphenylethylenediamine (8. 74g, 0.06 mole) and redist i l led triethylamine (7.07g, 0.07 mole) in 180 m l . dry ether were placed in the flask. The flask was i m m e r s e d in an ice-sal t bath for 1 hour until the temperature was between -5 and - 1 0 ° C . A solution of cyclohexanecarboxyl chloride (8. 742g, 0.06 m l . ) in 50 m l . ether was added very slowly f rom the dropping funnel with st i rr ing so as to keep the temperature of the reaction mixture below 0 ° C . After the addition was completed, s t i rr ing was continued at 0 ° C for another hour. The solution was st i rred overnight at room temperature. 30 m l . distil led water was then added, and the solution s t i rred rapidly for 10 minutes. The layers were separated, and the aqueous phase was extracted with two 100 m l . portions of ether. The extracts were combined with the ether layer, and dried over 50 g of anhydrous magnesium sulphate. The solvent was removed using a rotary evaporator and the residual product distil led under reduced pressure using a 9 inch Vigreux column. The fraction boiling point at 124-125 ° C (0. 15 m m . Hg. ) was collected. Y i e l d : I6g (92.4%). i r (neat) 2920, 2940, 2910, 2860 (C-H) , 1645 c m - 1 (C=0); nmr (CDCI3) % 7. 35 - 7.67 (m, 5, phenyl-H), 3.42 - 2.4 (m, 4, N - C H 2 - C H 2 - N ) , 4.47 - 3.93 (m, 1 ,CH-C=0) , 2.20 (s, 9 , ( o - C H 3 & N C C H 3 ) 2 ) , 2 . 0 - 0.67 (m, 10, C 6 H 1 Q ) . Mono-hydro chloride derivative The white salt was prepared by the same procedure as des-cribed in E x p . A - 3>: i m . p . 2 3 9 - 2 4 0 ° C . A n a l . C a l c d . for C l g H 2 9 N 2 C I O : C , 66. 54; H , 8.99; N , 8.62; C l , 10.92. Found: C , 66. 57; H , 8.92; N, 8.48; C l , 10.90. 5. N , N-dimethyl -N' - cyc lohexylmethyl -N' -o -methylphenyl -ethylene diamine A 250 m l . three necked flask was equipped with a mechanical 53 s t i r rer , dropping funnel (125 m l . ) and reflux condenser. The funnel and condenser car r ied drying tubes. D r y ether (180 m l . ) and .-. L A H \ ' (3. 64g, 0.096 mole) were placed in the flask. The solution was gently refluxed for 3 hours. A solution of N-o-methylphenyl -N-( ft -N , N-dimethylamino) cyclohexanecarboxamide (13.84g, 0.048 m l . ) in dry 40 m l . ether was very slowly added so as to maintain a gentle reflux. After the addition was completed, the mixture was s t i r red and refluxed for 20 hours. At the end of the reaction, the heating mantle was replaced by an ice-sal t bath, and the mixture s t i r red for 1 hour. 30 m l . disti l led water was added dropwise to the flask to decompose excess hydride. . Stirring was continued for 30 minutes until the mixture turned completely white. A sufficient amount of 40% sodium hydroxide solution was added to separate the ethereal layer f rom the ether-insoluble residue. The ethereal phase was decanted into an Er lenmeyer flask, and dried over anhydrous magnesium sulfate. The drying agent was suction filtered, and the solvent removed using a rotary evaporator. The residue was distil led under reduced pressure through a 9- inch Vigreux column warmed by a heating tape. The fraction boiling at 126r?128°C (0.4 m m . Hg) was collected. Y i e l d : 12. 75g (97%). i r (neat) 3010, 3050 ( A r y l - H ) , 2820, 2850 (N-(CH 3)^1470, 1495 c m ' 1 ( - C H 3 ) ; nmr (CDC13). £ 7. 30 - 6.87 (m, 4, phenyl-H), 3.30 - 2.73 (m, 4, - N - C H ^ C H ^ N ) , 2.3 (s, 3, 0 - C H 3 ) , 2.17 (s, 6, - N ( C H 3 ) 2 ) > 2 . 5 (m, 2, C H - C H 2 - N ) , 2.03 - 0.67 (m, 11, C 6 H n ) . Di -hydro chloride derivative The salt was prepared by the same procedure as described in Exp . A - 3 . A n a l . . Calcd . for C l g H 3 2 N 2 C l 2 : C , 62. 23; H , 9.27; N , 8.06; C l , 20.44. Found: C, 62.03; H , 9.28; N , 7.84; C l , 20.09. B . N . N - D I M E T H Y L - N ' - C Y C L O H E X Y L M E T H Y L - N ' - m - M E T H Y L -P H E N Y L E T HY L E N E D I A M I N E 1. °f -Chloro-N-m-methylphenylacetamide A solution of m-methylani l ine (32. l g , 0. 3 mole) and r e -distilled triethylamine (30. 34g, 0.3 mole) in 500 m l . dry ether was placed in 1 litre three-necked flask. The flask was equipped with a mechanical s t i r rer , a side a r m for setting a thermometer (range -100 to 5 0 ° C ) and a dropping funnel (125 m l . ) with a drying tube. The solution was cooled by st irr ing it for 1 hour in an ice-sal t bath (-5 to - 1 0 ° C ) . A solution of chloroacetylchloride (34g, 0.3 mole) in 100 m l . of dry ether was added slowly f rom the dropping funnel into the flask, with constant s t i r r ing, so as to keep the temperature of the solution below 0 ° C . The mixture was st i rred overnight at room temperature. Then 100 m l . of distil led water was added to the flask, and the mixture was st i rred for 15 minutes. The resulting white product in the mixture was suction filtered through a Buchner funnel. The ethereal layer was then separated and flash evaporated to remove the solvent. The remaining solids were combined with the suction filtered product in a 500 m l . beaker. 100 m l . each of 5% HC1 and cold distil led water were added and the product was vigorously s t i r red on a magnetic s t i r rer for 1 hour. The product was then suction f i l -tered and washed with distil led water until no HC1 could be detected using litmus paper. The product was predried in the air and finally dried at 4 0 ° C in an oven under reduced pressure (20 m m . Hg). The product was purified by reduced pressure sublimation. Y i e l d : 49. 0g (89%). m . p . : 9 2 ° C . i r (KBr) 3300 (0=C-N-H) , 3140, 3100, 2920 ( N ( C H 3 ) 2 ) , 1660 (C=0), 1610 (aromatic), 710 c m " 1 ( C - C l ) ; nmr ( C D C 1 3 ) % 7 . 4 7 -7 .13 (m, 4, phenyl-H), 4,15 (s, 2, C H 2 - C 1 . ) . 2.33 (s, 3, m - C H j ) . A n a l . Calcd . for C g H 1 Q N C I O : C, 58.9; H . 5.45; N , 7.64; C l , 19.35. Found: C , 59.00; H , 5. 59; N , 7.80; C l , 19.52. 2. c( -Dimethylamino-N-m-methyIphenylacetamide A 500 m l . three-necked flask was equipped with a dropping funnel (125 m l . ) fitted with a drying tube, a mechanical s t i r rer , and a side a r m for setting a thermometer (range -100 to 5 0 ° C ) . 300 m l . dry ether was placed in the flask and it was cooled to between -5 and - 1 0 ° C in an ice-sal t bath for 1 hour. Dimethylamine (26.5 cc, 0.4 mole) was trapped in a dry graduated cylinder (50 m l . ) from a dimethylamine gas tank using an acetone-dry ice bath ( - 8 0 ° C ) and poured all at once into the three-necked flask. A solution of 56 c\ -chloro-m -methylphenylacetamide flask (27. 6g, 0.15 mole) in 80 m l . dry tetrahydrofuran was added very slowly f rom the dropping funnel to the rapidly s t i r red solution at such a rate as to keep the temperature of the reaction mixture below 0 ° C for 10 hours. The mixture was kept overnight with st irr ing below 1 0 ° C . The white dimethylamine salt was suction filtered, and the solvent removed by a rotary eva-porator. 30 m l . distil led water was added to the product, and it was shaken vigorously in the separating funnel. The mixture was extracted twice using 200 m l . portions of ether, and the ethereal solution was dried over anhydrous magnesium sulfate. The solution was suction filtered and the solvent removed using a rotary evaporator. The product was redist i l led on oi l bath using a 9 inch Vigreux column under reduced pressure . Y i e l d : 27. 5g (95. 5%). b . p. : 1 09?C (0. 35 m m . Hgj . r i r (neat) 3300 (0=C-NH), 2940, 2820, 2780 (N(CH )_), 1675 (C=0), 1620, 1590 (aromatic), 790 c m " 1 (phenyl-H); nmr ( C D C l j ) & 7.43 - 6. 73 (m, 4, phenyl-H), 2.97 (s, 2, 0 = C - C H 2 ) , 2.30 (s, 6, ( N - C H ) 2), 2.27 (s, 3, m - C H 3 ) . Mono-perchlorate derivative The aminoacetamide (3. 56g, 0.02 mole), was. dissolved in 15 m l . of absolute ethanol. Then 3. 08 m l (0. 022 mole) of perchloric acid (70%) was added. The mixture reacted at room temperature for 15 minutes. The salt was crystal l ized out by the addition of ether. It was recrystal l ized from ethanol and dry ether, m . p . : 1 0 5 - 1 0 6 ° C . 57 A n a l . . Calcd . for C n H 1 7 N C I O : C, 45. 2; H , 5.82; N , 9.6; C l , 12.14. Found: C , 45. 27; H , 5.95; N , 9.44; C l , 12.03. 3. N, N-dimethyl-N'-m-methylphenylethylenediamine A dry 250 m l . three-necked flask was equipped with a mechanical s t i r rer , a dropping funnel (125 m l . ) and a reflux con-denser. The funnel and condenser both car r ied drying tubes. 20 m l . dry ether and lithium aluminum hydride (7.2g, 0.2 mole) were added to the flask, and the mixture was gently refluxed with s t i r r ing for 3 hours. A solution of o(-dimethylamino-N-m-methylphenylacetamide (19. 2g, 0. 1 mole) in 50 m l . dry ether was placed in the dropping funnel, and added dropwise so as to maintain gentle refluxing. After the addition was completed, the mixture was refluxed for 40 hours with stirring on the electr ical heating mantle. When the reduction was complete, the reaction mixture was cooled by placing it in an ice-sal t bath for 30 minutes. 20 m l . distil led water was added very slowly to the rapidly s t i r red mixture in the flask to decompose excess hydride. St irr ing was continued until the mixture became white in colour. Sufficient 40% N a O H solution was added to allow clear separation of the ethereal layer . The ether-insoluble residue was separated by centrifugation. . The ethereal layer was dried overnight using anhydrous magnesium sulfate. The solvent was removed with a rotary evaporator, and the residue distilled on an oi l bath under reduced pressure using a 9 inch Vigreux column wrapped with a heating tape. The fraction 58 boiling at 7 0 ° C (0.025 m m . Hg) was collected. Y i e l d : 16. ?g (95%). i r (neat) 3370 (N-H), 3040 (phenyl-H), 3030, 2930, 2850, 2810, 2760 ( - N ( C H 3 ) 2 ) , 1600 (aromatic), 770, 695 c m " 1 (phenyl-H); nmr (CDCI3) $ 7. 20 - 6.92 (m, 1, C & H 1 ) , 6.63 - 6. 30 (m, 3, C 6 H 3 ) , 4. 37 - 3.97 (weak s, 1, N - H ) , 3.30 - 2.90 (m, 2, N H - C H 2 ~ C ) 2.67 - 2. 37 (m, 2, ( N H - C - C H 2 ) , 2.28 (s, 3, m - C H 3 ) , 2.23 (s, 6, - N ( C H 3 ) 2 ) . Di -hydro chloride derivative The salt was prepared by the same procedure as described in Exp . A - 3 . m . p . : 1 6 3 ° C . A n a l . Calcd . for C ^ ^ 2 Q ^ 2 ^"12* C * 5 2 - 5 9 5 ; H « 8 - 0 3 5 N , 11.15; C l , 28. 22. . Found: C , 52. 57; H , 8.07; N , 11.01; C l , 28.12. 4. N - m - m e t h y l p h e n y l - N - ( (? - N , N-dimethylaminoethyl) cyclo- hexanecarboxamide A dry 250 m l . three-necked flask was equipped with a mechanical s t i r rer , dropping funnel (125 m l . ) with drying tube, and a side a r m for setting a thermometer (range -100 to 5 0 ° C ) . . N , N -dimethyl-N'-m-methylphenylethylenediamine (1 2. 45g, 0.07 mole) and redist i l led dry triethylamine (8.08g, 0.08 mole) in 170 m l . dry ether were placed in the flask. The flask was i m m e r s e d in an ice-salt bath for 1 hour until the temperature of the solution was between -5 and - 1 0 ° C . 50 m l . of a solution of cyclohexanecarbonylchloride (10. 12g, 0. 07 m l . ) in ether was added very slowly f rom the dropping 59 funnel so that the temperature of the reaction mixture remained below 0 ° C . After the addition was completed, the mixture was st i rred at 0 ° C for 1 hour. 30 m l . distil led water was added, and the mixture was s t i rred rapidly for 10 minutes at room temperature. The layers were separated, and the aqueous phase extracted with three 120 m l . portions of ether. The extracts were combined with the ethereal layer, and dried over 50g of anhydrous magnesium sulfate. The solvent was removed with a rotary evaporator and the residual product was distilled under reduced pressure using a 9 inch Vigreux column. The fraction boiling at 1 1 6 - 1 1 8 ° C (0.005 m m . Hg) was collected. Y i e l d : 17. 15g (85%). i r (neat) 2920, 2850, 2760 ( N ( C H 3 ) 2 ) , 1650 (C=0), 1600 c m " 1 , (aromatic); n m r - ( C D C l 3 ) £ 7.40 - 6.83 (m, 4, C 6 H^), 3.92 - 3.58 (m, 2, 0 = C - N - C H 2 ) , 2. 70 - 2.43 (m, 2, 0 = C - N - C - C H 2 ) , 2.70 -2.43 (m, 2, 0 = C - N - C - C H 2 ) , 2.35 (s, 3, m - C H ^ ) , 2.18 (s, 6, N ( C H 3 ) 2 ) . Mono-hydro chloride derivative The salt was prepared by the same procedure as described in E x p . A - 3 . l-m.p. : 2 3 1 - 2 3 2 ° C . A n a l . C a l c d . for C l g N 2 C>2 C l : C , 66.54; H , 8.99; N , 8. 62; C l , 10.92. Found: . C, 66. 70; H , 9 . 0 9 ; N , 8.47; C l , 10.92. 60 5. . N , N - d i m e t h y l - N ' - c y c l o h e x y l m e t h y l - N ' - m - m e t h y l p h e n y l - e thylenediamine A Z50 m l . three-necked flask was equipped with a mechanical s t i r rer , a reflux condenser and a dropping funnel (125 m l . ) . The con-denser and funnel both car r ied drying tubes. 180 m l . dry ether and lithium aluminum hydride (4. 18g, 0. 11 mole) were placed in the flask. The solution was gently refluxed for 3 hours on the heating mantle. A solution of jN'-m-methylphenyl-N^-C^ -N,N-dimethyiaminoethyl)cyclohexane carboxamide . .Iv (15. 8g, 0. 55 mole) in 40 m l . dry ether was added drop-wise to the refluxing L i A l H ^ solution f rom the dropping funnel in such a manner as to maintain gentle refluxing. After the addition was completed, the mixture was refluxed with st irring for 30 hours. At the end of the reaction, the heating mantle was replaced by an ice-salt bath, and 30 m l . of disti l led water was added dropwise into the flask to decompose excess hydride. St irr ing was continued for 30 minutes until the mixture turned white, and sufficient 40% N a O H solution was added to separate the ethereal layer f r o m the ether-insoluble residue. The ethereal layer was decanted and dried over anhydrous magnesium sulfate overnight. The drying agent was suction filtered, and the solvent removed using a rotary evaporator. The residue was distil led under reduced pressure through a 9 inch Vigreux column warmed by a heating tape. The fraction boiling at 1 4 0 ° C (0.4 m m . Hg) was collected. Y i e l d : 14. 5g (96%). ir (neat) 3050 (C-H) , 2920, 2860, 2770 ( p C - H 2 ) , 1600, 1580 61 (aromatic ring), 770 c m " 1 (phenyl-H): nmr ( C D C 1 3 ) & 7. 30 - 6.93 (m, 1, p h e n y l - o - H ) , 6.60 - 6.33 (m, 3, C 6 H 3 ) , 3.6 - 3.27 (m, 2, C H - C H 2 - N ) , 3 . 2 - 3.0 (d, 2,2=6, N C H 2 - C - N ) , 2 . 6 - 2 . 3 3 (m, 2, N - C - C H 2 - N ) , 2.30 (2s, 9, N ( C H 3 ) 2 ) , ( m - C H j ) , 2.0 - 0.67 (m, 11, C 6 H l l > -C . N , N - D I M E T H Y L - N ' - C Y C L O H E X Y L M E T H Y L - N ' - p - M E T H Y L -P H E N Y L E T HY L E N E D I A M I N E 1. °j -Chloro-N-p-methylphenylacetamide A solution of p-methylaniline (32. l g , 0. 3 mole) and disti l led triethylamine (30. 34g, 0.3 mole) in 500 m l . dry ether was placed in a 1 litre three-necked flask. The flask was equipped with a mechanical s t i r rer , a side a r m for setting a thermometer, (range -100 to 5 0 ° C ) and a dropping funnel (125 m l . ) with a drying tube. The solution was cooled by s t i r r ing it for 1 hour in an ice bath. A solution of chloro-acetylchloride (134g, 0.3 mole) in 100 m l . dry ether was added from the dropping funnel into the flask. The solution was added slowly with s t i rr ing, so as to keep the temperature of the solution below 0 ° C . The mixture was s t i r red overnight at room temperature. Then 100 m l . of distilled water was added to the flask, and the mixture s t i r red for 15 minutes. The resulting white product was suction fil tered through a Buchner funnel. The ethereal layer was separated and flash evaporated to remove the solvent. The product was placed in a 500 m l . beaker and 100 m l . each of 5% HC1 and distil led water were added. The mixture was vigorously s t i rred on a magnetic s t i r rer for one hour. The product was then suction filtered and washed with distil led water until no HC1 could be detected using litmus paper. The product was p r e -dried in the air and finally dried at 4 0 ° C in an oven under reduced pressure (20 m m . Hg). The product was purified by reduced pressure sublimation. Y i e l d : 50.6g(91%). m . p . : 1 6 0 ° C . i r (KBr) 3300 (0=C-NH) 2950, 2940, 2860 ( C - H ) , 1665 (C=0) 1610 (aromatic, 730 c m ' 1 ( C - C l ) ; nmr ( C D C 1 3 ) S 7. 55 - 7.02 (m, 4, phenyl-H), 4. 17 (s, 2, C H ^ - C l ) , 2.32 (s, 3, p - C H j ) . A n a l . . C a l c d . for H Q N C I O : C, 58.9; H , 5.45; N , 7.64; C l , 19.32. Found: C, 58.96; H , 5 .62 ; .N, 7.74; C l , 19.39. 2. o( -Dimethylamino-N-p-methylphenylacetamide A 500 m l . three-necked flask was equipped with a dropping funnel (125 m l . ) fitted with a drying tube, a mechanical s t i r rer , and a side a r m for setting a thermometer (range- 1 0 0 ° C to 5 0 ° C ) . 300 m l . of dry ether was placed in the flask and cooled to between - 5 ° and - 1 0 ° C in an ice-sal t bath for 1 hour. Dimethylamine (approx. I6.55cc, 0.25 mole) was trapped in a dry 50 m l . graduated cylinder f rom a dimethylamine gas tank using an acetone-dry ice bath ( - 8 0 ° C ) . It was poured all at once into the three-necked flask. A solution of e | -chloro-N-p-methylphenylacetamide (18.36g, 0.1 mole) in 60 m l . dry tetrahydrofuran was added slowly f rom the dropping funnel to the 63 vigorously s t i r red solution so as to keep the temperature of the reaction mixture below 0 ° C . When the addition was completed, the mixture was st i rred for 10 hours at a temperature below 0 ° C for 10 hours, and finally s t i r red overnight at a temperature below 1 0 ° C . The white dimethylamine salt was suction filtered and the solvent removed using a rotary evaporator, then 20 m l . distilled water was added to the product, and the mixture was shaken vigorously in the separatory funnel. It was extracted twice with 150 m l . portions of ether, and the ethereal solution was dried over anhydrous magnesium sulfate. The solution was suction filtered and the solvent removed with a rotary evaporator. The product was redist i l led on an oil bath using a 9 inch Vigreux column under reduced pressure . Y i e l d : 17. 3g (90%). b . p . : 1 0 3 - 1 0 5 ° C (0. 10 m m . Hg). ir (neat) 3280 (0=C-NH), 2930, 2830, 2770 ( N ( C H 3 ) 2 ) , 1660 (C=0), 1580 c m " 1 (aromatic); nmr ( C D C l ^ ) , £ 7 . 5 8 - 7 . 0 (m, 4, phenyl-H), 3.03 (s, 2, C H 2 - N ) , 2.35 (s, 6, N ( C H 3 ) 2 > 2.27 (s, 3, p - C H 3 ) . 3. N , N-dimethyl-N'-p-methylphenylethylenediamine A dry 250 m l . three-necked flask was equipped with a mechanical s t i r rer , dropping funnel (125 m l . ) and reflux condenser. The funnel and condenser both car r ied drying tubes,. 120 m l . dry ether and lithium aluminum hydride (5. 7g, 0.15 mole) were placed in the flask and gently refluxed with st i rr ing for 3 hours. A solution of 64 «( -dimethylamino-N-p-methylphenylacetamide (14g, 0.75 mole) in 50 m l . dry ether was placed in a dropping funnel and added dropwise at such a rate as to maintain gentle reflux. When the addition had been completed, the mixture was refluxed for 40 hours with st irr ing on the electr ical heating mantle. After the reduction was over, the reaction mixture was cooled in an ice-sal t bath for 30 minutes, and 20 m l . distil led water was added dropwise to the rapidly s t i r red mixture in the flask to decompose excess hydride. . Stirring was continued until the mixture became white in colour, then sufficient 40% N a O H solution was added to allow clear separation of the ethereal layer . The ether-insoluble residue was separated by centrifugation. The ethereal layer was dried over anhydrous magnesium sulfate over-night. The solvent was removed using a rotary evaporator and the residue was further distil led under reduced pressure . The fraction boiling at 6 5 - 6 8 ° C (0.025 m m . Hg) was collected. Y i e l d : 12.75g (95.5%). i r (neat) 3360 (N-H), 2930, 2850, 2810, 2760 (N(CH ) 2), 1610 c m - 1 (aromatic); nmr ( C D C 1 3 ) , 5 7 . 1 3 - 6 . 4 (m, 4, phenyl-H), 4. 17 - 3. 73 (broad s, 1, N - H ) , 3. 22 - 2. 87 (q, 4, J=6, N H - C H 2 ) , 2. 60 - 2.30 (q, 4, J = 6, - C H 2 - N ( C H 3 ' ) 2 ) , 2.22 (s, 6, ( N ( C H 3 ) 2 ) . A n a l . . C a l c d . for C l g N 2 C I O : C, 66. 54; H , 8. 99; N , 8.62; C l , 10. 92. . Found: C , 66. 32; H , 8.94; N , 8.52; C l , 10 .91. 65 4. N - p - m e t h y l p h e n y l - N - ( ^ - N t N - d i m e thy lamino ethyl)- cyclohexanecarboxamide A dry 250 m l . three-necked flask was equipped with a mechanical s t i r rer , dropping funnel (125 m l . ) with a drying tube, and side a r m for setting a thermometer (range -100 to 5 0 ° C ) . N , N-dimethyl -N' -p_-methyl -phenylethylenediamine (10. l g , 0.06 mole) and redist i l led by triethylam-ine (10. l g , 0. 06 mole) in 170 m l . dry ether were placed in the flask. The flask was cooled to between -5 and - 1 0 ° C in an ice-sal t bath for one hour with vigorous s t i r r ing . 50 m l . ethereal solution of cyclohexane carboxyl-chloride (8. 742g, 0.06 mole) was added slowly from the dropping funnel so as to keep the temperature of the reaction mixture below 0 ° C . After the addition was completed, s t irr ing was continued at 0 ° C for another hour. Then the mixture was s t i r red overnight at room temperature. 30 m l . distil led water was added, and the mixture was st i rred rapidly for 10 minutes. The layers were separated and the aqueous phase was extracted with three 120 m l . portions of ether. The extracts were combined with the ethereal layer, and dried over 50g of anhydrous magnesium sulfate. The solvent was removed using a rotary evaporator under reduced p r e s -sure using a 9 inch Vigreux column. The fraction boiling at 1 2 8 - 1 3 0 ° C (0. 125 m m . Hg) was collected. Y i e l d : 15.4g(89%). i r (neat) 3040, 730 (aryl-HO, 2920, 2840, 2810, 2760 ( N - ( C H ) 2), 1650 (C=0), 1600, 1580 c m " 1 (aromatic); nmr ( C D C 1 3 ) , & 7.4-6.93 (m, 4, phenyl-H), 3. 93 - 3. 60 (m, 2, 0 = C - N - C H 2 ) , 2. 60 - 2. 22 (m, 2, - C H 2 ~ N ( C H 3 ) 2 ) , 2.20 (s, 6, ( N ( C H 3 ) 2 ) . 1.87 - 0.85 (m, 11, C 6 H u ) . Mono-hydro chloride derivative The salt was prepared by the same procedure as described in E x p . A - 3 : m . p . : 2 4 0 ° C . A n a l . - C a l c d . for C l 8 H 2 g N 2 C I O : C , 6 6 . 5 4 ; H , 8 . 9 9 ; N , 8.62; C l , 10.92. Found: C, 66. 32; H , 8. 94; N , 8.52; C l , 10.91. 65a •5. . N , N-dime thy 1 - N ' - cyclohexylmethyl-N' -p-methylphenyl- ethylenediamine A Z50 m l . three-necked flask was equipped with a mechanical s t i r rer , reflux condenser and dropping funnel (125 m l . ) . The funnel and condenser both car r ied drying tubes. 180 m l . dry ether and lithium aluminum hydride (3. 8g, 0. 1 mole) were placed in the flask. The solution was gently refluxed on the heating mantle f o r 3 hours. A solution of N-p-methylphenyl-N-( . -dimethylaminoethyl) cyclohexane-carboxamide (14. 4g, 0.05 m l . ) in 40 m l . dry ether was added f rom the dropping funnel so as to maintain gentle reflux. After the addition was completed, the mixture was refluxed for 30 hours with s t i r r ing . At the end of the reaction, the heating mantle was replaced by an ice-sal t bath and st i rred for 1 hour. 20 m l . disti l led water was added dropwise to the flask to decompose excess hydride. . St irr ing was continued for 30 minutes until the colour of the mixture turned completely white. A solution was added to separate the ethereal layer f rom the ether-insoluble residue. The ethereal phase was decanted into an Er lenmeyer flask and dried over anhydrous magnesium sulfate. The drying agent was suction filtered, and the solvent removed using a rotary evaporator. The residue was distil led under reduced pressure through a 9 inch Vigreux column warmed by a heating tape.. The fraction boiling at 1 3 8 ° C (0. 4 m m . Hg) was collected. Y i e l d : 13. 3g (97%). The ir spectrum of unpurified product showed the absence of the C=0 band at 1650 c m - 1 , indicating complete reduction of the amide. 66 D . . S Y N T H E S I S O F N , N - D I M E T H Y L - N ' - C Y C L O H E X Y L M E T H Y L - N ' -p - B R Q M O P H E N Y L E T H Y L E N E D I A M I N E 1. ^ -Chloro-N-p-bromophenylacetamide A 1 litre three-necked flask was equipped with a mechanical s t i r rer , side a r m for setting a thermometer (range - 1 0 0 ° - 5 0 ° C ) , and dropping funnel (1Z5 m l . ) with a drying tube. A solution of p-bromoaniline (5. I6g, 0.3 mole) and distil led triethylamine (30. 3g, 0. 3 mole) in 50 m l . dry ether were added to the flask. The solution was cooled to between -5 and - 1 0 ° C in an ice-salt bath with st i rr ing for 1 hour. A solution of chloroacetyl chloride (34. Og, 0.3 mole) in 100 m l . dry ether was added very slowly to the vigorously s t i rred solution from the dropping funnel so as to keep the temperature of solution below 0 ° C . After the addition was completed, the mixture was st irred at room temperature overnight, then 100 m l . distilled water was added, and the mixture was st irred for 15 minutes. The white product in the mixture was suction filtered through a Buchner funnel, and the layers were separated. The ethereal layer was flash evaporated to remove the solvent and the solid was combined with the suction filtered product. The product was poured into a 500 m l . beaker and 100 m l . each of 5% HC1 solution and cold distil led water were added to it . The mixture was s t i r red virogou&ly on a magnetic s t i r rer for 1 hour. The product was suction filtered, and washed with 300 m l . distil led water until no trace of HC1 could be detected using litmus paper. The product was dried in the air first , and then in the oven under reduced pressure (40 C , 20 m m . Hg). The product was purified by reduced pressure sublimation on the oil bath. Y i e l d : 67. l g (90%). m . p. : 1 8 2 ° C . i r - ( K B r ) 3260 (0=C-NH), 1660 (C=0). 1600, 1540, (aromatic), 780 ( C - C l ) , 750 c m " 1 (phenyl-Br) ; nmr (CDC1 3 ) 8 7.43 (m, 4, phenyl-H), 4. 15(s, 2, C H 2 - C 1 ) , 1.50 (s, 1, N - H ) . A n a l . . C a l c d . for C g H ? N B r C I O : C, 38.6; H , 2.82; N , 5.63; C l , 14. 3; B r , 32.2. Found: C , 38.69; H , 2.93; N , 5.72; C l , 14. 22; B r , 32. 04. 2. o( -Dimethylamino-N-p-bromophenylacetamide A 500 m l . three-necked flask was equipped with a dropping funnel (250 m l . ) with a drying tube, mechanical s t i r re r , and a side a r m for setting a thermometer (range - 1 0 0 ° C - 5 0 ° C ) . 150 m l . each of dry ether and dry tetrahydrofuran were added to the flask. The solution was cooled to between -5 and - 1 0 ° C in an ice-sal t bath for 1 hour. Dimethylamine (26.5 cc,- 0.4 mole) was trapped in a dry 50 m l . graduated cylinder f r o m a dimethylamine gas tank using an acetone-dry ice bath ( - 8 0 ° C ) and poured al l at once into the flask. A solution of o f -chloro-N-p-bromophenylacetamide (37. 2g, 0.15 mole) in 150 m l . dry ether was added slowly f rom the dropping funnel to the vigorously s t i r red solution so as to keep the temperature of the reaction mixture below 0°;C.. . T h e ' m i x t u r e was then st i rred for 10 68 hours while being kept below 0°C, and finally s t i r red overnight while being kept below 1 0 ° C . The white dimethylamine salt was suction filtered and the solvent was removed using a rotary evaporator, then 30 m l . distilled water was added to the product, and the mixture was shaken vigorously in the separating funnel. The mixture was extracted twice with two 200 m l . portions of ether, and the ethereal solution was dried over anhydrous magnesium sulfate. The solution was suction filtered and the solvent removed using rotary evaporator. The product was recrysta l l ized f rom a dry mixture consisting of hexane and a small amount of ethylacetate. Y i e l d : 31. 6g (82%). m . p . : 5 7 - 5 8 ° C . i r (KBr) 3260 (0=C-NH), 2970, 2940, 2820, 2770 ( N ( C H 3 ) 2 ) , 1660 (C=0) , 750 c m " 1 ( C - B r ) ; nmr ( C D C 1 3 ) & 7.43 (m, 4, phenyl-H), 3.0 (s, 2, 0 = C - C H 2 ) , 2.33 (s, 6, N(CH 3 ) 2 . ) . 1.57 (s, 1, N - H ) . A n a l . . Calcd . for C-^Q H ^ 3 N 2 B r O : C, 46. 75; H , 5.06; N , 10.90; B r , 31.18. Found: C , 46.62; H , 5.13; N , 10.80; B r , 31.08. 3. . N , N-dimethyl-N'-p-bromophenylethylenediamine A dry 250 m l . three-necked flask was equipped with a mechanical s t i r rer , dropping funnel (125 m l . ) and reflux condenser. The condenser and dropping funnel both car r ied drying tubes. 120 m l . dry ether and lithium aluminum hydride (7. 6g, 0. 2 mole) were added to the flask and gently refluxed with s t i r r ing for 3 hours . A solution of «>< -dimethylamino-N-p-bromophenylacetamide (25. 7g, 0. 1 mole) 69 in 180 m l . dry ether was placed in a dropping funnel and added drop-wise so as to maintain gentle reflux. After the addition was completed, the mixture was refluxed for 40 hours with s t i r r ing . When the reduc-tion was over, the reaction mixture was cooled in an ice-sal t bath for 30 minutes, then 20 m l . distil led water was added dropwise to the rapidly s t i r red mixture in the flask to decompose excess hydride. St irr ing was continued until the mixture became white in colour. Sufficient 40% N a O H was added to allow clear separation of the ethereal layer f rom the solids. The ether-insoluble residue was separated by centrifugation. . The ethereal layer was dried over anhydrous magnesium sulfate overnight. The solvent was flash evaporated, and the residue disti l led under reduced pressure using a heating tape on the oil bath and a 9 inch Vigreux column. The fraction boiling 9 0 ° C (0. 05 m m . Hg) was collected. Y i e l d : 23.08g(95%). i r (neat) 3370 (N-H), 3040 (aryl-H) , 2930, 2850, 2805, 2760 ( N ( C H 3 ) 2 ) , 1600 c m - 1 (aromatic); nmr (CDC1 3) S 7.33 - 6. 33 (m, 4, .phenyl-H), 3.33 - 2.90 (m, 2, N H - C H 2 ) , 2. 65 - 2.30 (m, 2, C H 2 ~ N ( C H 3 ) 2 ) , 2.25 (s, 6, N ( C H 3 ) 2 ) . D i - h y d r o chloride derivative The salt was prepared by the same procedure as described in Exp. A - 3 . m . p . : 1 3 0 ° C . A n a l . C a l c d . for C l Q H j ? N 2 .Br C l : C, 37. 99; H , 5.42; 70 N , 8.86; C l , 22.46;-Br, 25.27. / F o u n d : C, 38.16; H , 5.47;. «N-, 8.84; C l , 22. 30; B r , 2.5. 20. 4. N - p - b r o m o p h e n y l - N - ( - N , N-dimethylaminoethyl)- cyclohexanecarboxamide A dry 250 m l . three-necked flask was equipped with a mechanical s t i r rer , a dropping funnel (125 m l . ) with a drying tube, and side a r m for setting a thermometer (range - 1 0 0 ° C - 5 0 ° C ) . N , N-dimethyl-N'-p-bromophenylethylenediamine (19.44g, 0.08 mole) and redist i l led dry triethylamine (9.1g, 0.09 mole) in 170 m l . dry ether were added to the flask. The flask was cooled to between - 5 ° and - 1 0 ° C in an ice-sal t bath for 1 hour. 50 m l . ethereal solution of cyclohexanecarboxyl chloride (11.68g, 0.08 mole) was added very slowly f rom the dropping funnel to the rapidly s t i r red contents of the flask, so as to keep the temperature of the reaction mixture below 0 ° C . After the addition was completed, s t i rr ing was continued at 0 ° C for 1 hour, and then overnight at room temperature. 30 m l . of distil led water was added, and the mixture was st i rred rapidly for 10 minutes. The layers were separated, and the aqueous phase was extracted with three 120 m l . portions of ether. . The extracts were combined with the ether layer, and dried over 50g of anhydrous magnesium sulfate. The solvent was removed using a rotary evaporator and the residual product was recrysta l l ized using petroleum ether ( 3 0 ° C - 6 0 ° C ) . Y i e l d : 22. 5g (80%). m . p . : 8 l - 8 2 ° C . "71 i r (neat) 3040 (phenyl-H), 2920, 2850, 2760 ( N ( C H 3 ) 2 ) • 1640 (C=0), 750 c m " 1 ( C - B r ) ; nmr (CDCI3) 5 7. 65 - 6.97 (m, 4, phenyl-H), 3.90 - 3.60 (m, 2, 0 = C - N - C H 2 ) , 2.53 - 2 . 2 0 (m, 2, - C H ^ N ( C H 3 ) 2 ) , 2 . l 6 ( s , 6, N ( C H 3 ) 2 ) . A n a l . Calcd . for C 1 ? H 2 5 N 2 O B r : C, 57.93; H , 7.09; N , 7.95; B r , 22.68. Found: C, 57.61; H , 7. 08; N , 8. 00; B r . 22.43. 5. N , N - d i m e t h y l - N ' - c y c l o h e x y l m e t h y l - N ' - p - b r o m o p h e n y l -ethylene diamine A 500 m l . three-necked flask was equipped with a mechanical s t i r rer , dropping funnel (125 m l . ) and reflux condenser. The funnel and condenser both car r ied drying tubes. 30 m l . dry ether and lithium aluminum hydride (3. 8g, 0. 1 mole) were placed in the flask. The solution was gently refluxed for 3 hours on the heating mantle. A solution of N , p-bromophenyl-N-( ft - N , N-dimethylaminoethyl) cyclohex-anecarboxamide (24. 6g, 0.07 mole) in 50 m l . dry ether was added very slowly to the flask so as to maintain gentle reflux. After the addition was completed, the mixture was refluxed for 12 hours with s t i r r ing . The heating mantle was then replaced by an ice-sal t bath and the mixture was s t i rred for 1 hour. 30 m l . of distil led water was added dropwise to the cooled reaction mixture, with vigorous st i rr ing, to decompose excess hydride. Stirr ing was continued for half an hour. Next, suffi -cient 40% N a O H solution was added to separate the ethereal layer, which was decanted into an Er lenmeyer flask and dried over anhydrous magnesium sulfate. The drying agent was suction filtered, and the solvent removed using a rotary evaporator. The residue was disti l led under reduced pressure through a 9 inch Vigreux column warmed by an electrical heating tape. The fraction boiling at 1 5 0 ° C (0.2 m m . Hg) was collected to yield tertiary amine (14.88g, 62.85%), and 6.35 g of the secondary amine decomposition product of the amide, c o r r e s -ponding to 8. 78g (37. 34%) of tertiary amine product. i r (neat) 3040 (phenyl-H), 2920, 2850, 2820, 2780 ( N ( C H 3 ) 2 ) 1590 (aromatic), 750 c m " 1 ( C - B r ) ; nmr ( C D C 1 3 ) S 7. 30 - 7.10 (m, 2, H at C 3 & C 5 of phenyl), 6. 60 - 6. 37 (m, 2, H at C 2 & C 6 of phenyl), 3.55 - 2.92 (m, 4, C H 2 - N - C H 2 ) , 2.58 - 2. 28 (m, 2, C H 2 - N ( C H 3 ) 2 ) , 2.23 (s, 6, N ( C H 3 ) 2 ) , 2. 0 - 0.67 (m, 11, C f e H ). Di -hydro chloride derivative The salt was prepared by the same procedure as described in Experiment A - 3 . m . p . : 1 8 3 ° C . ' A n a l . Calcd . for C ^ H N ' 2 B r C l 2 : C , 49. 52; H , 7. 086; N, 6.79; C l , 17.22; B r , 19.28. Found: C, 49.58; H , 7. 1 9 ; N , 6.94; C l , 17. 05; B r , 19. 55. E . SYNTHESIS O F N , N - D I M E T H Y L - N 1 - C Y C L O H E X Y L M E T H Y L - 3, 5-D I M E T H Y L P H E N Y L E T H Y L E N E D I A M I N E 1. °( - C h l o r o - N - 3 , 5-dimethylphenylacetamide A one litre three-necked flask was equipped with a mechanical s t i r rer , side a r m for setting a thermometer (range -100Q\C - 50° C), 73 and dropping funnel (125 m l . ) with a drying tube. A solution of 3. 5 dimethylaniline (24g, 0.2 mole) and triethylamine (20.24g, 0.2 mole) in 400 m l . dry ether was placed in the flask. The solution was cooled to between -5 and - 1 0 ° C in an ice-sal t bath with st irr ing for 1 hour. A solution of chloroacetyl chloride (22. 6g, 0. 2 mole) in 70 m l . dry ether was added very slowly to the vigorously s t i rred solution f rom the dropping funnel so as to keep the temperature of the solution below 0 ° C . After the addition was completed, the mixture was st i rred at room temperature overnight. 100 m l . distilled water was added to the flask and the mixture was st i rred for 15 minutes. The white product in the mixture was suction filtered through a Buchner funnel and the layers were separated. . The ethereal layer was flash evaporated to remove the solvent and the solid was combined with the suction filtered product. The product was poured into a 500 m l . beaker and 100 m l . each of 5% H C l and cold distil led water were added to it. The product was st i rred vigorously on a magnetic s t i r rer for one hour. The product was suction filtered, and washed with 300 m l . distilled water until no trace of H C l could be detected with litmus paper. The product was dried in the air , and then in the oven under reduced p r e s -sure ( 4 0 ° C , 20 m m . . Hg). It was purified by reduced pressure sub-limation on an oi l bath. Y i e l d : 36g(91.5%). m . p . : 1 4 0 ° C . i r (KBr) 3300 (0=C-NH) , 3220, 3160, 3120, 2960, 2980, 2860 ( C H 2 , C H 3 ) , 1660 (C=0), 1615 (aromatic), 645 c m " 1 ( C - C l ) ; nmr ( C D C 1 3 ) § 7.30 - 7.06 (m, 2, phenyl -H at & C 6 ), 6.80 (m, 1, 74 phenyl -H at C 4 ) , 4.13 (s, 2, C H - C l ) . 2.27 (s, 6, 3. 5-dimethyl). A n a l . Calcd . for C 1 0 H 1 2 N O C l : C, 60. 7; H , 6.07; N , 7.08;. C l , 17 .98. . Found: C, 60. 74; H , 6. 12; N , 7.27; C l . 18.09. 2. °( - D i m e t h y l a m i n o - N - 3 , 5-dimethylphenylacetamide A 500 m l . three-necked flask was equipped with a dropping funnel (125 m l . ) with drying tube, a mechanical s t i r rer , and a side a r m for setting a thermometer (range f rom - 1 0 0 ° C - 5 0 ° C ) . 150 m l . each of dry ether and dry tetrahydrofuran were placed in the flask. The solution was cooled to between -5 and - 1 0 ° C in an ice-sal t bath for 1 hour. Dimethylamine (approx. 8. Occ, 0.3 mole) was trapped in a dry graduated cylinder f rom a dimethylamine gas tank using an acetone-dry ice bath ( - 8 0 ° C ) , and poured all at once into the three-necked reaction flask. A solution of ^ - c h l o r o - N - 5 , 6-dimethylphenyl-acetamide (23. 8g, 0. 13 mole) in 80 m l . dry tetrahydrofuran was added very slowly f rom the dropping funnel-to the vigorously s t i r red solution so as to keep the temperature of the reaction mixture below 0 ° C . After the addition had been completed, the mixture was kept below 0 ° C for 10 hours arid st irred overnight. The.white dimethylamine salt was suction filtered and the solvent removed using a rotary evaporator. 30 m l . distilled water was added to the product, and it was shaken vigorously in the separating funnel. The mixture was extracted twice with 150 m l . portions of ether, and the ethereal solution was dried over anhydrous magnesium sulfate. The solution was suction fil tered and the 75 solvent was removed by flash evaporation. i r (neat) 3290 ( C O N H ) , 2930, 2820, 2770 ( N ( C H 3 ) 2 ) , 1600 c m " 1 (aromatic); nmr ( C D C l ^ ) S 7.20 (s, 2, phenyl-H at C 2 and C^), 6.70 (s, 1, phenyl -H at C 4 ) , 3.00 (s, 2, C - C H 2 ) , 2. 33 (s, 6, N ( C H 3 ) 2 ) . . Mono-perchlorate derivative The salt was prepared by the same procedure as described in Exp. B - 2 . m . p . : 1 8 8 ° C . A n a l . C a l c d . f o r C ^ H N ^ . O : C, 47.0; H , 6. 20; N , 9.14; C l , 11.58. Found: C , 47.13; H , 6.31; N , 9.25; C l , 11.52. 3. . N , N-dimethyl -N|-3 , 5-dimethylphenylethylenediamine A dry 250 m l . three-necked flask was equipped with a mechanical s t i r rer , dropping funnel (125 m l . ) and reflux condenser. Both the funnel and the condenser car r ied drying tubes. 150 m l . dry ether and lithium aluminum hydride (6. 45g, 0. 17 mole) were placed in the flask and gently refluxed with st i rr ing for 3 hours. A solution of °1 -d imethylamino-N-3 , 5-dimethylphenylacetamide (17g, 0. 085 mole) in 50 m l . dry ether was placed in a dropping funnel and added dropwise at such a rate as to maintain a gentle reflux. When the addition was completed, the mixture was refluxed for 40 hours with st irr ing on the electr ical heating mantle. After the reduction had been completed, the reaction mixture was cooled in an ice-sal t bath for 30 minutes and 20 m l . of distil led water was added dropwise to the rapidly s t i rred mixture in the flask to decompose excess hydride. Stirring was continued until the 76 mixture became white in colour and then sufficient 10% N a O H solution was added to facilitate separation of the ethereal layer . The ether-insoluble residue was separated by centrifugation and the ethereal layer was dried over anhydrous magnesium sulfate overnight. The solvent was removed with a rotary evaporator, and the residue was distilled under reduced pressure using a 9 inch Vigreux column and a heating tape on the oil bath. The fraction boiling a t ' 7 0 ° C (0.05 m m . Hg) was collected. i r (neat) 3370 (N-H), 2930, 2850, 2810, 2760 (N(CH 3 ) 2 . ) , 1600 c m " 1 (aromatic); nmr ( C D C 1 3 ) 5 6.40 - 6.20 (d, 3, J=4, phenyl-H), 4.30 - 3.90 (broad 's , 1, N - H ) , 3.50 - 2.87 (m, 2, N H - C H 2 ) , 2.63 - 2.30 (m, 2, C H 2 - N ( C H 3 ) 2 ) . 2.23 (s, 6, 3, 5-dimethyl), 2.17 (s, 6, N ( C H 3 ) 2 ) . D i - h y d r o chloride derivative The salt was prepared by the same procedure as described in Exp. A - 3 . . m . p . : 1 8 8 ° C . - A n a l . C a l c d . for C j 2 H 2 2 N 2 C l . C, 54. 33; H , 8. 35; N , 10.56; C l , 26.76. . Found: 54. 39;• H, 8 . 2 3 ; N , 10.45; C l , 26.83. 4. . lNf-3, 5-dimethylphenyl-N-( ft - N , N-dimethylamino)-cyclohexane-carboxamide A dry 250 m l . three-necked flask was equipped with a mechanical s t i r rer , a dropping funnel (125 m l . ) with drying tube, and a side a r m for setting a thermometer (range -100 to 5 0 ° C ) . . N , N -d i m e t h y l - N ' - 3 , 5-dimethylphenylethylenediamine (1.5g, 0.02 mole), redisti l led dry triethylamine (7. l g , 0. 7 mole) and 170 m l . dry ether were added to the flask. The flask was cooled to between -5 and - 1 0 ° C in an ice-salt bath for one hour. 50 m l . of a solution of cyclohexane-carboxyl chloride (8. 742g, 0. 06 mole) in ether was very slowly added to the rapidly s t i r red mixture f rom the dropping funnel so as to keep the temperature of the mixture below 0 ° C . When the addition had been completed, s t irr ing was continued at the same temperature for another hour, and then overnight at room temperature. 20 m l . of distil led water was added and the mixture was s t i r red rapidly for 10 minutes. The layers were separated, and the aqueous phase was extracted with three 120 m l . portions of ether. . The extracts were combined with the ether layer, and dried over anhydrous magnesium sulfate. The solvent was removed with a rotary evaporator. The product was distilled under reduced pressure using a 9 inch Vigreux column. The fraction boiling at 1 4 5 ° C (0.4 m m . Hg) was collected. . Y i e l d : 14. 6g (81%). ir (neat) 2920, 2840, 2760 ( N f C H ^ ) , 1645 c m " 1 (C=0); nmr ( C D C 1 3 ) S 7. 03 - 6. 73 (d, 3, J= 10. phenyl-H), 3. 93 - 3. 60 (m, 2, 0 = C - N - C H 2 ) , 2.57 - 2.30 (m, 8, 3 ,5-dimethyl & C H 2 ( N C H ^ ) , 2.33 (s, 6, 3. 5-dimethyl) 2.22 (s, 6, N(CH 3 ) 2 > , 2.0 - 0.40 (m, 11, C 6 H u ) . Mono-perchlorate derivative The salt was prepared by the same procedure as described in E x p . A - 3 . m . P . : 1 9 3 - 1 9 4 ° C . F . - N , N - D I M E T H Y L - N 1 - C Y C L O H E X Y L M E T H Y L - N ' - C Y C L O H E X Y L -E T HY L E N E D I A M I N E 1. . Cyclohexanecarboxyl chloride A one litre three-necked flask was equipped with a reflux condenser and a dropping funnel. . Both the funnel and condenser carr ied drying tubes. Cyclohexanecarboxylic acid (128g, 1 mole) was placed in the flask and thionyl chloride (179g, 1. 5 mole) was added over a period of 5 minutes with s t i r r ing . The flask was placed in an oi l bath and heated at a bath temperature of 1 5 0 ° C for 1 hour. The reflux condenser was then replaced by a distillation head. 200 m l . dry benzene was added and the mixture was distil led until the tempera-ture of the vapour reached 9 5 ° C . The mixture was then cooled and another 200 m l . of anhydrous benzene was added. The distillation was continued until the temperature of the vapour again reached 9 5 ° C . The residual acid chloride was redist i l led under reduced pressure on the oil bath to give a colourless l iquid. Y i e l d : 120g (82%). b . p . : 4 6 ° C (2. 75 m m . Hg). (Lit . ) b . p . : 67-67. 5 ° C (14 m m . Hg). i r (neat) 2920, 2850 ( C - H 2 ) , 1770 (C=0), 790 c m - 1 ( C - C l ) . 2. °{ -Chloro-N-cyclohexylacetamide A 2 l i tre three-necked flask was equipped with a mechanical s t i r rer , side a r m for setting a thermometer (range -100 - 5 0 ° C ) and 250 m l . dropping funnel with a drying tube. A solution of cyclohexyl-amine (198. 1 8g, 2 mole) in 1. 5 1. dry ether was placed in the flask. 79 The solution was st i rred over a dry ice-acetone bath for 30 minutes until it reached - 3 0 ° C . A solution of chloroacetyl chloride (112. 95g, 1 mole) was added dropwise to the vigorously s t i r red solution from the dropping funnel so as to keep the temperature below - 3 0 ° C . After the addition was completed, the st irr ing was continued in the ice-sal t bath for 3 hours, and overnight at room temperature. 200 m l . 5% HC1 solution was added to the reaction mixture, which was then s t i r red for 5 minutes. The white product which precipitated was suction fil tered and the ethereal layer was separated from the acidic aqueous phase with a separatory funnel. The filtered product was washed with distilled water until no trace of acid could be detected on.litmus paper. The ethereal layer was washed twice with 200 m l . portions of distilled water, and dried over anhydrous magnesium sulfate overnight. The solvent was removed with a rotary evaporator to recover more amide. The product was predried in the air and then sublimed under reduced pressure . Y i e l d : 158g(90%). m . p . : 1 0 5 ° C . ir (KBr) 3220 (0=C-NH), 3023, 2875, 2800 ( C H 2 ) , 1630 (C=0), 790 c m " 1 ( C - C l ) ; nmr ( C D C 1 3 ) § 6. 92 - 6. 00 ( s , 1, N H ) , 4. 03 (s, 2, C H 2 - C 1 ) , 3.43 (m, 1, C H - N ) , 2.40 - 0.50 (m, 10, C 6 H 1 Q ) . A n a l . C a l c d . for C„ H , . N C l O: C, 54.65; H , 7.97; N , 7.97; — — — o 14 C l , 20.2. Found: C , 54. 56; H , 8.15; N , 7.84; C l , 20.21. 80 3. o( - D i m e t h y l a m i n o - N - cyclohexylacet amide A 2 litre three-necked flask was equipped with a dropping funnel (500 m l . ) with drying tube, mechanical s t i r rer , and side a r m for setting a thermometer (range - 1 0 0 ° C - 5 0 ° C ) . 500 m l . dry ether and 200 m l . dry tetrahydrofuran were placed in the flask. . The solution was cooled to below 0 ° C in an ice-sal t bath for 1 hour. Dimethylamine (approx. 122. 5cc, 1.7 mole) was trapped in a dry graduated cylinder from a dimethylamine gas tank using an acetone-dry ice bath ( - 8 0 ° C ) and poured into the flask al l at once. A solution of o| - c h l o r o - N -cyclohexylacetamide (149g, 0.85 mole) in 400 m l . dry T H F was added slowly from the dropping funnel to the vigorously s t i rred solution so as to keep the temperature below 0 ° C . After the addition was completed, the solution was kept below 0 ° C and st irred for 10 hours, and then kept below - 1 0 ° C and st i r red overnight. . The white dimethylamine salt was suction filtered off and the solvent removed by flash evaporation. 30 m l . distil led water was added to the product and it was shaken vigorously in the separatory funnel. The mixture was extracted twice with 200 m l . portions of ether, and the ethereal solution was dried over anhydrous magnesium sulfate. The solution was suction filtered and the solvent removed by flash evaporation. The crude product was r e c r y s -tallized from a mixture of ether, petroleum ether and hexane. Y i e l d : 176. 8g (96%). m . p . : 6 l ° C . ir (KBr) 3260 (0=C-NH), 2960 - 2760 (N(CH 3 )J , 1630 c m " 1 81 (C=0); nmr ( C D C 1 3 ) 5 4. 0 - 3. 57 (broad s, 1, N - H ) , 2. 90 (s, 2, C H 2 - N ) , 2.27 (s, 6, N ( C H 3 ) 2 ) , 2.10 - 0.80 (m, 11, C 6 H n ) . A n a l . . Calcd . for C 1 Q H 2 Q N 2 O : C, 65.2;. H , 10.87; N , 15.21. Found: C, 64.99; H , 10. 74;. N , 15.33. 4. . N , N - d i m e t h y l - N ' - eye lohexylethylenediamine A 2 litre three-necked flask was equipped with a 500 m l . dropping funnel, reflux condenser, and mechanical s t i r r e r . The funnel and condenser both car r ied drying tubes. One 1. dry ether was placed in the flask and lithium aluminum hydride (51. 4g, 1. 35 mole) was added. The mixture was gently refluxed with st irr ing for 3 hours. A solution of o| -dimethylamino-N-cyclohexylactamide (165. 5g, 0.9 m l . ) in 250 m l . dry ether was added to the flask so as to maintain gentle reflux. After the addition was completed, the mixture was refluxed for 40 hours with s t i r r ing. When the reduction was over, the flask was cooled in an ice-sal t bath. Excess lithium aluminum hydride was decomposed by dropwise addition of distilled water until the mixture turned white. - Sufficient 40% N a O H was added to give clear separation of the ethereal phase from the ether-insoluble residue. The ethereal solution was dried over anhydrous magnesium sulfate overnight. The ether was removed by flash evaporation and the product was distil led under reduced pressure . Y i e l d : I45g (95%). b . p . : 7 3 ° C ( 2 . 0 m m . Hg). ir (neat) 3300 (NH), 2920, 2860, 2840, 2780 c m " 1 . ( N ( C H 3 ) 2 J ; 82 nmr ( C D C 1 3 ) 8 3. 00 - 2.30 (m, 4, N - C H 2 - C H 2 - N ) , 2.20 (s, 6, N { C H 3 ) 2 ) , 2.05 - 0.7 (m, 12, C 6 H n , NH) . Di -hydro chloride derivative The salt was prepared by the same procedure as described in E x p . A - 3 . m . p . : 2 3 1 - 2 3 2 ° C . A n a l . Calcd . for C j 0 H 2 4 N 2 C l 2 : C , 49.3; H , 9 . 8 ; N , 11.5; C1-, 29.2. . Found: C, 49.29; H , 9.77;. N, 11.54; C l , 29.39. 5. N - c y c l o h e x y l - N - ( P - N , N-dimethylaminoethyl)-cyclohexane-carboxamide A 250 m l . three-necked flask was equipped with a mechanical s t i r rer , and a dropping funnel (125 m l . ) with a drying tube. N , N -dimethyl-N'-cyclohexylethylenediamine (5.1g, 0.03 mole) and dry triethylamine (4. 04g, 0.4 mole) in 120 m l . dry ether were placed in the flask. The solution was cooled to between -5 and - 1 0 ° C in an ice-sal t bath for one hour with s t i r r ing . A solution of cyclohexanecarboxyl chloride (4. 35g, 0.03 mole) in dry ether (40 m l . ) was added very slowly from the dropping funnel to the vigorously s t i rred solution so as to keep the temperature of the solution below 0 ° C . After the addition was c o m -pleted, the s t i r r ing was continued at room temperature overnight, 20 m l . of distil led water was added and the mixture was st i rred for 10 minutes. The ether layer was separated f rom the aqueous layer using a separatory funnel. . The aqueous phase was extracted twice with 80 m l . portions of ether. The extracts were combined with the ether 83 layer and dried over anhydrous magnesium sulfate. The drying agent was suction filtered and most of the solvent was removed using a rotary evaporator. The crude product was distil led under reduced pressure through a 9 inch Vigreux column, warmed by a heating tape. The fraction boiling at 1 3 5 ° C (0.1 m m . Hg) was collected. The liquid product solidified at room temperature. Y i e l d : 7. 04g (84%). ir (KBr) 2940, 2860, 2840, 2785 ( -N(CH 3 ) 2 . ) , 1644 c m " 1 (C=0); nmr ( C D C 1 3 ) S 3.60 - 2.20 (m, 11, C H - N - C H 2 - C H 2 - N ( C H 3 ) 2 ) , 2.32 (s, 6, N ( C H 3 ) 2 ) , 2.20 - 0.87 (m, 21, C & H n , C & H 1 Q ) . . Mono-hydro chloride derivative The salt was prepared by the same procedure as described in E x p . A - 3 . m . p . : 1 9 7 ° C . A n a l . Calcd . for C 1 ? H N 2 ' O C l : C, 64.4; H , 10.4; N , 8.8; C l , 11.2. Found: C, 64. 27; H , 1 0 . 3 1 ; N , 8.91; C l , 11.43. 6. . N , N-dimethyl -N' -cyclohexylmethyl -N' -cyclohexylethylene- di amine A dry 250 m l . three-necked flask was equipped with a mechanical s t i r rer , dropping funnel (125 m l . ) and refluxing condenser. Both the funnel and condenser carr ied drying tubes. 120 m l . dry ether was placed in the flask and lithium aluminum hydride (0. 76g, 0. 04 mole) was added to it. The solution was gently refluxed with st irring for 3 hours. A solution of N - c y c l o h e x y l - N - ( ^ - N , N-dimethylaminoethyl) -N-84 cyclohexanecarboxamide (5.715g, 0.0219 mole) in 30 m l . dry ether was added dropwise from the funnel to the L i A l H ^ solution, so as to maintain gentle reflux. After the addition was completed, the mixture was refluxed with st irr ing for 30 hours. The heating mantle was then replaced by an ice-sal t bath, and the solution was cooled below 0 ° C . 10 m l . of distil led water was added dropwise to the rapidly s t i r red mixture in the flask to decompose excess hydride. St irr ing was con-tinued for 20 minutes. Sufficient 40% N a O H solution was added to bring about clear separation of the ethereal layer . The mixture was centr i -fuged and the ethereal layer was dried over anhydrous magnesium sulfate overnight. The drying agent was suction fil tered and the solvent removed using a rotary evaporator. The crude product was distil led under reduced pressure through a 9 inch Vigreux column. The fraction boiling at 1 6 0 ° (2 m m . Hg) was collected. . Y i e l d : 5. 6g (96%). i r (neat) 2920, 2850, 2760 ( N ( C H 3 ) 2 ) ; nmr ( C D C l j ) S 2.70 -2.08 (m, 11, C H - N - C H 2 - C H 2 - N ( C H 3 ) 2 ) , 2.27 (s, 6, N ( C H 3 ) 2 ) , 2 . 1 0 -0.42 (m, 23, C & Hn C H 2 , C 6 H 1 Q ) . Di -hydro chloride derivative The salt was prepared by the same procedure as described in E x p . A - 3 . m . p . : 2 1 9 - 2 2 0 ° C . •Anal . Calcd . f o r C 1 ? H 3 6 N 2 C l 2 : C, 60.1; H , 10.6; N , 8.2; C l , 20.9. Found: C, 59.91; H , 10.68;. N , 8.09; C l , 20.74. 85 G . SYNTHESIS O F N . N - D I M E T H Y L - N ' - B E N Z Y L - N ' - C Y C L O H E X Y L -E T HY L E N E D I A M I N E 1. . N - c y c l o h e x y l - N - ( ft - N , N-dimethylaminoethyl)-phenyl-carboxamide A 250 m l . three-necked flask was equipped with a mechanical s t i r re r , dropping funnel with drying tube, and a side a r m for setting a thermometer (range - 1 0 0 ° C - 5 0 ° C ) . A solution of N , N - d i m e t h y l - N ' -cyclohexylethylenediamine (6. 8g, 0.04 mole) and dry triethylamine (V. 5g, 0. 05 mole) in 150 m l . dry ether was placed in the flask. The flask was i m m e r s e d in an ice-sal t bath for 1 hour until the temperature of the solution was below 0 ° C . A solution of benzoyl chloride (5.6g, 0. 04 mole) in 50 m l . dry ether was added dropwise to the flask f r o m the dropping funnel with st i rr ing, so as to keep the temperature of the reaction mixture below 0 ° C . After the addition was completed, s t i r r ing was continued at 0 ° C for another hour, and at room temperature over-night. 50 m l . of disti l led water was added, the layers were separated and the aqueous phase was extracted with two 100 m l . portions of ether. The extracts were combined with the ether layer, washed with saturated sodium chloride solution and dried over 50g anhydrous magnesium sulfate. The solvent was removed by flash evaporation and the residual liquid distil led under reduced pressure using a 9 inch Vigreux column. The fraction boiling at 1 5 5 ° C - 1 6 0 ° C (0,5 m m . Hg) was collected, and the product was recrystal l ized f rom a pentane-hexane mixture. Y i e l d : 8.98g (82%). m . p . : 5 6 ° C . 86 ir (neat) 2930, 2860, 2820, 2770 ( N ( C H 3 ) 2 ) , 1630 c m - 1 (C=0); nmr ( C D C 1 3 ) S 7.33 (m, 5, phenyl-H), 3.63 - 3.23 (m, 2, 0 = C - N -C H 2 ) , 2. 68 - 0.50 (m, 19, C & H j 1 & C H 2 - N ( C H 3 ) 2 ) . A n a l . C a l c d . for C 1 7 H 2 & N 2 ' 0 : . C, 74. 5; H , 9. 5; N , 10.2. Found: C, 74.29; H , 9.34; N', 10.05. 2. N , N-dimethyl -N' -benzyl -N' -cyclohexylethylenediamine A 250 m l . three-necked flask was: equipped with a dropping funnel (125 m l . ) , reflux condenser, and mechanical s t i r r e r . The con-denser and funnel both car r ied drying tubes. 175 m l . dry ether and lithium aluminum hydride (1.248g, 0. 032 mole) were added to the flask, and gently refluxed with s t i r r ing for 3 hours on the electric heating mantle. A solution of N - c y c l o h e x y l - N ' - ( ^ - N , N-dimethylaminoethyl)-benzenecarboxamide (4. 38g, 0.016 mole) in 25 m l . dry ether was placed in the dropping funnel, and added very slowly so as to maintain gentle reflux. When the addition was completed, the mixture was st irred and refluxed for 40 hours. After the reduction was over, the reaction flask was cooled in an ice-sal t bath. Excess lithium aluminum hydride was decomposed by the slow addition of distilled water with vigorous st irring so as to avoid violent reaction. A sufficient amount of 40% N a O H solution was added to give good separation of the ethereal layer f rom the ether-insoluble residue, and the ethereal solution was dried over anhydrous magnesium sulfate overnight. The ether was removed by flash evaporation and the residue was distil led under 87 reduced pressure . Y i e l d : 4. 25g (97%). ir (neat) 3040 (phenyl-H), 2920, 2850, 2820, 2760. ( N ( C H 3 ) 2 ) , 1600 c m " 1 (aromatic); nmr ( C D C 1 3 ) 8 7. 42 - 7.02 (m, 5, phenyl-H), 3.60 (s, 2, p h e n y l - C H 2 ) , 2.77 - 2.17 (m, 4, N - C H 2 - C H 2 - N ) . 2.12 (s, 6, N - ( C H 3 ) 2 ) , 2.00 - 0.70 (m, 11, C & H ). Di-perchlorate derivative The salt was prepared by the same procedure as that described in Exp . B-2 , except that two equivalents of perchloric acid was used, m . p . : 9 4 ° C . A n a l . . Ca lcd . for C 1 ? H 3 Q N 'Og. C l 2 : C , 44. 40; H, 6. 50; N , 6.09; C l , 15.08. Found: C, 44.13; H , 6.47; N, 5.97; C l , 15.23. H . N , N - D I M E T H Y L - N - D I P H E N Y L M E T H Y L E T HY L E N E D I A M I N E 1. ^ - Chip ro-N-diphenylme thy lac etamide A 250 m l . three-necked flask was equipped with a mechanical s t i r rer , a side a r m for setting a thermometer (range f rom - 1 0 0 ° C -5 0 ° C ) , and a dropping funnel (125 m l . ) with drying tube. Diphenyl-methylamine (54. 9g, 0.3 mole), redist i l led dry triethylamine (40.48g, 0.4 mole) and 150 m l . dry ether were placed in the flask. The solution was s t i rred in an ice-sal t bath until the temperature of the solution reached -5 to - 1 0 ° C . A solution of chloroacetyl chloride (33. 9g, 0. 3 mole) in 40 m l . dry ether was added very slowly to the rapidly s t i r red solution in the flask from a dropping funnel so as to keep the 88 temperature of the reaction mixture below 0 ° C . When the addition had been completed, the st irr ing was continued for 3 hours below 0 ° C and overnight at room temperature. 50 m l . of distilled water was added and the mixture was s t i r red for 15 minutes. . The white product in the mixture was suction filtered through a Buchner funnel, and the layers were separated. The ethereal layer was flash evaporated to remove the solvent, and the solids were combined with the suction filtered product. The product was poured into a 500 m l . beaker and 100 m l . each of 5% HC1 solution acid and cold distilled water was added to it. The mixture was st i rred vigorously on a magnetic s t i r rer for 1 hour, then suction filtered and the product was washed with 400 m l . distilled water until no trace of HC1 could be detected with litmus paper. The product was dried in the air and then in an oven under reduced pressure ( 4 0 ° C , 20 m m . Hg). The amide was recrystal l ized from a mixture of dry petroleum ether ( 3 0 ° - 6 0 ° C ) and ethyl acetate. Y i e l d : 71. 5g (92%). m . p . : 1 2 8 - 1 2 9 ° C . i r (KBr) 3200 (N-H), 3040 (phenyl-H) 1650 c m " 1 (C=0); nmr ( C D C l 3 ) i 7 . 28 (sharp m , 10, di-phenyl-H) 408, s , 2 , ( 0 = C - C H 2 ) , 1. 50 (s, 1, C H - N ) . > • A n a l . Calcd . for C , , H , N O C 1 : C, 69.4; H , 5.4; N , 5.4; 1 3 14 C l , 13. 70. . Found: C , 69.29; H , 5.44; N , 5.53; C l , 13.54. 89 2. ck -dimethylamino-N-diphenylmethylacetamide A dry 500 m l . three-necked flask was equipped with a mechanical s t i r rer , a side a rm for setting a thermometer ( - 1 0 0 ° C -5 0 ° C ) and a dropping funnel (250 m l . ) with a drying tube. 100 m l . each of dry ether and tetrahydrofuran were placed in the flask. The solution was cooled to between -5 and - 1 0 ° C in an ice-sal t bath for 1 hour. Dimethylamine (39.72 m l . , approx. 0.6 mole) was trapped in a dry graduated cylinder f rom a dimethylamine gas tank using an acetone-dry ice bath ( - 8 0 ° C ) and was poured into the reaction flask. A solution of o| -chloro-N-diphenylmethylacetamide (74. 1 g, 0.285 mole) in 150 m l . tetrahydrofuran was added very slowly f rom the dropping funnel to the rapidly s t i r red reaction mixture so as to keep the temperature below 0 ° C . After the addition was completed, the mixture was st i rred for 10 hours below 0 ° C and then overnight below 1 0 ° C . The white dimethyl-amine salt was suction filtered and the solvent was removed using a rotary evaporator. 40 m l . distil led water was added to the product and it was shaken vigorously in the separating funnel. The mixture was extracted with three 150 m l . portions of ether, and the ethereal solution was dried over anhydrous magnesium sulfate. The solution was suction filtered and the solvent was removed with a rotary evaporator. The product was recrys ta l l ized f rom a mixture of hexane and small amount of ethyl acetate. Y i e l d : 68g (89%). m . p . : 8 3 ° C . i r (KBr) 3330 (0=C-N-H) , 2780 - 3060 (N(CH )2) 1670 c m " 1 90 (C=C, C=0); nmr ( C D C 1 3 ) § 7 . 4 - 7.18 (m, 10, di -phenyl-H) , 6.37 - 6.17 (d, 1, J = 9 Hz C H - N ) , 2.97 (s, 2, 0 = C - C H 2 - N ) , 2.20 (s, 7, N ( C H 3 ) 2 ) . A n a l . C a l c d . for C 1 ? H 2 Q N ^ O : C, 76.1; H , 7.46; N , 10.45. Found: C , 76.10; H , 7. 57; N , 10.29. 3. . N , N-dimethyl-N'-diphenylmethylethylenediamine A one litre three-necked flask was equipped with a mechanical s t i r rer and a Soxhelt apparatus connected to a reflux condenser with a drying tube. 600 m l . dry ether and lithium aluminum hydride (19. 0g, 0. 5 mole) were placed in the flask. The solution was gently refluxed for 3 hours with s t i r r ing . -dimethylamino-N-diphenylmethylaceta-mide (55g, 0. 242 mole) was packed in the round-bottomed filter paper column of the Soxhelt extractor. A glass rod was inserted in the packed powder in order to give the solvent easy access to the powder. The reaction mixture was refluxed for 48 hours with gentle stirring on the oi l bath. At the end of the reaction, the oi l bath was replaced by an ice-sal t bath and the reaction mixture was cooled for 1 hour. 20 m l . disti l led water was added dropwise to the rapidly s t i r red mixture in the flask to decompose excess hydride. . Stirring was continued until the mixture turned white. Sufficient 40% sodium hydroxide solution was added to allow clear separation of the ethereal layer . The ether-insoluble layer was separated by decantation. The ethereal layer was dried over anhydrous magnesium sulfate overnight with s t i r r ing . The magnesium sulfate was suction filtered and the solvent removed using a rotary evaporator. The residue was distil led under vacuum on the oi l bath using a 9 inch Vigreux column wrapped with a heating tape. The fraction boiling at 1 3 5 ° C (0.4 m m . Hg) was collected. Y i e l d : 58. 5g (95%). Di-perchlorate derivative The salt was prepared by the same procedure as that describe in Exp . B -2 , except that two equivalents of perchloric acid was used, m . p . : 1 8 4 - 1 8 5 ° C . A n a l . . C a l c d . for C H N O C l : C, 44.84; H , 5.31; 17 24 2 8 2 N , 6.15; C l , 15.59. Found: C , 45.01; H , 5. 38; N , 6.27; C l , 15.46. I. N , N - D I M E T H Y L - N * . N ' - D I B E N Z Y L E T H Y L E N E D I A M I N E 1. N ' - b e n z y l - N ' - ( ft - N , N-dimethylaminoethyl)-benzene- carboxamide A dry 250 m l . three-necked flask was equipped with a mechanical s t i r rer , dropping funnel (125 m l . ) with a drying tube, and side a r m for setting a thermometer (range - 1 0 0 ° C - 5 0 ° C ) . N , N -dimethyl-N'-benzylethylenediamine (17.83g, 0.1 mole) and redist i l led triethylamine (10. l g , 0. 1 mole) in 180 m l . dry ether were placed in the flask. The solution was cooled to between.-5 and - 1 0 ° C in an i c e -salt bath for 1 hour. A solution of benzoyl chloride (14. l g , 0.1 mole) in 30 m l . dry ether was added very slowly from the dropping funnel to the rapidly s t i r red solution so that the temperature of the solution remained below 0 ° C . After the addition was completed, the st irr ing 92 was continued at room temperature overnight. 30 m l . disti l led water was added and the mixture was s t i r red for 10 minutes. The ethereal layer was separated from the aqueous one using a separatory funnel. The aqueous phase was extracted twice with 100 m l . portions of ether. These extracts were combined with the ether layer and dried over anhydrous magnesium sulfate overnight. The drying agent was suction filtered and the solvent was removed using a rotary evaporator. The residual crude product was distil led under reduced pressure through a 9 inch Vigreux column wrapped in a heating tape. The fraction boiling at 1 6 0 ° C (0.20 m m . Hg) was collected. . Y i e l d : 26. 8g (95%). ir (neat) 2940, 2830, 2780 ( N ( C H 3 ) 2 ) , 1632 c m " 1 (N-C=Q); nmr ( C D C T 3 ) 7. 60 - 7. 20 (m, 10, 2 phenyl), 4. 70 (s, 2, phenyl-C H 2 ) , 3.67 - 3.20 (m, 2, 0 = C - N - C H 2 ) , 2.67 - 1.92 (m, 8, C H 2 - N ( C H 3 ) 2 ) . Mono-hydro chloride derivatives The salt was prepared by the same procedure as that described in Exp . A - 3 . m . p . : 1 6 8 ° C . A n a l . Calcd . for C l g H 2 3 N 2 0 C l : . C, 67.8; H , 7.21; N , 8.78; C l , 11.12. Found: C, 67.68; H , 7.18;. N , 8.74; C l , 11.01. 2. N , N - d i m e t h y l - N ' , N'-dibenzylethylenediamine A dry 250 m l . three-necked flask was equipped with a mechanical s t i rrer and dropping funnel ( 125 m l . ) with a drying tube. 93 150 m l . dry ether was placed in the flask and lithium aluminum hydride (3.8g, 0.1 mole) was added to it. The solution was gently refluxed with st irr ing for 3 hours. A solution of N ' - b e n z y l r N ' - ( ft -N , N-dimethylaminoethyl) -benzenecarboxamide (14. 1.2g, 0. 05 mole) in 50 m l . dry ether was added very slowly to the flask from the dropping funnel so as to maintain gentle reflux. After the addition had been completed, the mixture was refluxed with st i rr ing for 30 hours. The heating mantle was replaced by an ice-water bath and the solution was cooled below 0 ° C . 20 m l . of distil led water was added dropwise to the rapidly s t i rred mixture in the flask to decompose excess hydride. Sufficient 40% N a O H was added to bring about clear separation of the ethereal layer . The mixture was centrifuged and the ether layer dried over anhydrous magnesium sulphate overnight. The drying agent was suction filtered and the solvent removed using a rotary evaporator. The crude product was distil led under reduced pressure using a 9 inch Vigreux column warmed by a heating tape. The fraction boiling at 1 4 3 ° C ( 0. 75 m m . Hg) was collected. . Y i e l d : 12. 7g (95%). ir (neat) 3040 (aryl-H) , 2940, 2800 ( N ( C H 3 ) 2 ) , 1600 c m " 1 (aromatic); nmr ( C D C 1 3 ) & 7 . 5 0 - 7.17 (m, 10, diphenyl-H), 3.61 (s, 4, 2 p h e n y l - C H 2 ) , 2.73 - 2.31 (m, 4, N - C H 2 - C H 2 - N ) , 2.17 (m, 6, N ( C H 3 ) 2 ) . Di -pic ra te derivative 3. l g of the tertiary amine was dissolved in 10 m l . of °94 absolute ethanol and 5 m l . saturated picr ic acid solution was added. The mixture was warmed for 15 minutes. The salt was recrysta l l ized from dry ethanol and ether m . p . : Z10. •Anal. C a l c d . for C , n H N O : C, 49.5; H , 4.15; H , 15.42. JU 30 o 14 Found: C, 48.97; H , 4.04; N , 15.58. J . . N , N - D I M E T H Y L - N 1 - 1 - CY C L O H E X E N Y L M E T H Y L - N 1 -CY C L O H E X Y L E T HY L E N E D I A M I N E 1. Cyclohexylcyanohydrin' A 500 m l . three-necked flask was equipped with a dropping funnel and mechanical s t i r r e r . Cyclohexanol (90. Og, 1 mole) in 100 m l . ether and finely powdered potassium cyanide (97. 6g, 1.5 mole) were placed in the flask in the fume hood. The flask was cooled in an i ce -salt bath for 2 hours with rapid s t i r r ing . A small excess of fuming H C l (150 cc) was added dropwise to the vigorously s t i r red reaction mixture and the stirring was continued for 10 hours. The inorganic salt was suction filtered, and the layers were separated. The ethereal layer was dried over anhydrous sodium sulfate. The drying agent was suction filtered and the ether solvent was distilled using a flash evaporator. The oily residue was distil led under reduced pressure in the fume hood and recrystal l ized from cyclohexane. Y i e l d : 90g (72%), m . p . : 3 1 ° C . ir (neat) 3420 (OH), 2950, 2870 ( C H 2 ) , 2260 c m " 1 (C=N); nmr ( C D C 1 3 ) S 3.03 (s, 1, O - H ) , 2.4 - 0.67 (m, 10, C & H 1 Q ) . A n a l . Calcd . for C . H n N O : C, 67.2; H , 8.80; N , 11.2. ——— o 11 Found: C, 67.09; H , 8.81; N , 11.39. 2. Methyl 1 -hydroxy-cyclohexanecarboxylate Cyclohexylcyanohydrin'^ (87. 5g, 0. 7 mole) and 500 m l . dry methanol were placed in a one litre round-bottomed flask equipped with a drying tube. The solution was cooled below 0 ° C in an ice-sal t bath for 2 hours. D r y HC1 gas was bubbled into the cooled reaction mixture until it had been saturated. . The solution was protected f rom moisture by the drying tube and left to stand overnight in the fume hood. The drying tube was replaced by an efficient reflux condenser. . The solution was slowly warmed and refluxed on the heating mantle for 5 hours in the fume hood. After the reaction was completed, the ammonium chloride was suction filtered and the methanol flash evaporated. 30 m l . disti l led water was added and the solution was s t i r red . The mixture was then extracted with three 150 m l . portions of ether and the ethereal solution was washed with 100 m l . • saturated sodium carbonate solution and dried over anhydrous magnesium sulfate. The drying agent was suction filtered and the solvent removed using a rotary evaporator. The residue was distilled under reduced pressure . The fraction boiling at 7 1 ° C (3 m m . Hg) was collected. . Y i e l d : 70g (63.4%). (Lit . b . p . : 9 3 - 9 8 ° C / l 3 m m . . Hg). 9 6 ir (neat) 3540 (O-H), 2970, 2890 ( C - H 2 ) , 1735 c m - 1 ( - C O - O ) ; nmr ( C D C 1 3 ) o" 3. 77 (s, 3, C H j ) , 3. 20 (s, 1, OH), 2. 0 - 0. 92 (m, 10, C 6 H 1 Q ) . A n a l . . C a l c d . for C g H 1 4 0 3 : C, 60.8; H , 8.85. Found: C, 60.93; H , 8. 83. 3. Methyl 1 - cyclohexenecarboxylate Methyl 1 -hydroxycyclohexanecarboxylate (55. 3g, 0. 352 mole) was placed in a 250 m l . three-necked flask equipped with a mechanical s t i r rer , dropping funnel (125 m l . ) and reflux condenser. The condenser and the funnel both car r ied calcium chloride drying tubes. The flask was placed in a cold water bath and st irr ing was begun. Thionyl chloride (83. 4g, 0. 74 mole) was added very slowly f rom the dropping funnel over a period of about an hour and the mixture was st i rred for 3 hours on the hot water bath ( 7 5 ° C ) . The excess thionyl chloride was removed by co-disti l lation with two 80 m l . portions of dry benzene. F inal ly , the residue was distil led under reduced pressure and the fraction boiling at 4 6 ° C (0.6 m m . Hg) was collected. Y i e l d : 42g (78.25%). ir (neat) 2950, 2880 ( C H 2 & C H 3 ) , 1715 ( - C O - O - ) , 1650 c m - 1 (C=C); nmr ( C D C 1 3 ) 5 7 . 1 3 - 6.77 (m, 1, C=CH), 3.73 (s, 3, 0 - C H 3 ) , 2.42 - 1.40 (m, 8, C & Hg). A n a l . Calcd . for Cg H 1 2 0 2 = C, 69.0; H , 9.00; Found: C, 68.47; H , 8.41. 4. 1 - cyclohexenecarboxylic acid Sodium hydroxide (12g, 0.3 mole) and 120 m l . disti l led water were placed in a 250 m l . three-necked flask fitted with a dropping funnel and reflux condenser. The solution was st i rred with a magnetic s t i r rer and 35g of methyl 1-cyclohexenecarboxylate was slowly added. When the^ester had been added, the solution was boiled gently for 3 hours until hydrolysis was complete ( i .e . , until a test portion could be completely dissolved in water). The mixture was diluted with 100 m l . distil led water, and about 100 m l . of liquid was removed by distillation to ensure that alcohol formed during the hydrolysis had been completely removed. 130 m l . 10% HC1 solution was added to the cold residue in the flask with s t i r r ing until the solution showed acidity with litmus paper. The upper organic acid layer was separated, and the aqueous phase extracted with three 150 m l . portions of ether. . The acid layer was combined with the ether extracts, washed with 40 m l . distil led water, and dried over anhydrous magnesium sulphate. The ether was flash evaporated arid the residue disti l led under reduced pressure . Y i e l d : 22g (87.3%). b . p . : 8 0 ° C / 2 m m . Hg. ir (neat) 3200 - 2500 ( -COOH), 1670 (-C=0), 1635 c m " 1 (C=C); nmr ( C D C 1 3 ) $ 4.17 (m, 1, C=CH), 2.5 - 1.3 (m, 8, C & H g ) . A n a l . Calcd . for C ? H 1 Q C>2: C , 66.60; H , 7.90. . Found: C, 66.50; H , 7. 94. 98 5. 1-cyclohexenecarboxyl chloride A dry 125 m l . three-necked flask was equipped with a reflux condenser, dropping funnel (125 m l . ) and mechanical s t i r r e r . The funnel and condenser both car r ied drying tubes. Cyclohexenecarboxylic acid (16. 4g, 0. 13 mole) was placed in the flask. Thionyl chloride (23. 8 g, 0.2 mole) was added over a period of 5 minutes to the acid, which was being s t i rred by a mechanical s t i r r e r . The flask was placed in an oi l bath at 1 5 0 ° C for one hour with rapid s t i r r ing . The reflux condenser was then replaced by a distillation head, 60 m l . of anhydrous benzene was added, and the mixture was distilled until the temperature of vapours reached 9 0 ° C . The mixture was allowed to cool, another 60 m l . of anhydrous benzene was added, and the distillation was con-tinued until the temperature of the vapours again reached 9 0 ° C . The residual acid chloride was distil led under reduced pressure to avoid pyrolysis of the acid chloride. Y i e l d : 13. 8g (74%). b . p . : 5 0 ° C (6 m m . Hg). i r (neat) 3030 ( = C - H ) , 1790 (C=0), 1651 (C=C), 650 c m " 1 ( C - C l ) . 6. N - c y c l o h e x y l - N - ( fi - N , N-dimethylaminoethyl)-1 - cyclo-hexenecarboxamide A 250 m l . three-necked flask was equipped with a mechanical s t i r re r , side a r m for setting a thermometer (range -100 to 5 0 ° C ) , and a dropping funnel (125 m l . ) with a drying tube. N , N - d i m e t h y l - N ' - c y c l o -99 hexylethyldiamine .(13 . 6g,._ 0,. 08 . mole) and triethylamine (15.18g, 0. 15 mole) in 150 m l . dry ether were placed in the flask. The solution was st irred in an ice-sal t bath for one hour until the temperature dropped below 0 ° C . A solution of 1 -cyclohexene-1-carbonylchloride (11.53g, 0. 08 mole) in 50 m l . dry ether was added very slowly f r o m the dropping funnel to the vigorously s t i r red solution so as to keep the temperature of the solution below 0 ° C . After the addition was completed, the reaction was allowed to continue at 0 ° C for 2 hours and then overnight at room temperature. 50 m l . of distil led water was added, the layers were separated, and the aqueous phase was extracted with two 100 m l . portions of ether. The extracts were combined with the ethereal layer, washed with saturated sodium chloride solution, and dried over 100 g of anhydrous magnesium sulfate. Most of the solvent was removed by flash evapora-tion, and the residual liquid was distil led under reduced pressure using a 9 inch Vigreux column. Y i e l d : 15.9g (74%). b . p . : 1 6 5 ° C (5 m m . Hg). ir (neat) 2920, 2850, 2770 ( N C C H 3 ) 2 ) , 1630 c m " 1 (C=C, C = 0 ) . / K . N, N - D I M E T H Y L - N ' - 3 - C Y C L O H E X E N Y L M E T H Y L - N 1 - C Y C L O -H E X Y L E T H Y L E N E D I A M I N E 1. 3-Cyclohexenecarboxyl chloride A 250 m l . three-necked flask was equipped with a reflux con-denser and dropping funnel, both of which carr ied drying tubes. 3 -cyclo-hexenecarboxylic acid (63g, 0. 5 mole) was placed in the flask. Thionyl chloride (95. 184g, 0.8 mole) was added over a period of 5 minutes to the 100 acid with s t i r r ing . The flask was placed in an oil bath and heated at a bath temperature of 1 5 0 ° C for one hour. The reflux condenser was then replaced by a distillation head, 150 m l . of anhydrous benzene was added, and the mixture was distil led until the temperature of the vapours reached 9 5 ° C . The mixture was cooled and another 150 m l . of anhydrous benzene was added. Distil lation was continued until the temperature of the vapours again reached 9 5 ° C . The residual acid chloride was redist i l led under reduced pressure, and the fraction boiling at 9 0 ° C (20 m m . Hg) was collected. Y i e l d : 56. 34g (78%). ir (neat) 3050 (=C-H), 1800 (0=C-CT), 655 c m " 1 ( C - C l ) . 2. . N-cyclohexyl-3-cyclohexenecarboxamide A dry 250 m l . three-necked flask was equipped with a mechanical s t i r rer , a dropping funnel (125 m l . ) with a drying tube, and a side a r m for setting a thermometer (range -100 - 5 0 ° C ) . A solution of cyclohexylamine (40g, 0.4 mole) in 300 m l . dry ether was placed in the flask. The solution was cooled to between -5 and - 1 0 ° C in an i ce -salt bath for one hour. A solution of 3-cyclohexenecarboxyl chloride (28.92g, 0.2 mole) in 60 m l . dry ether was added dropwise f rom the funnel to the rapidly s t i rred solution so as to keep the temperature of the solution below 0 ° C . After the addition was completed, the st irr ing was continued for one hour at the same temperature, and overnight at room temperature. 50 m l . of distil led water was added and the mixture was st irred for 20 minutes. The white solid was suction filtered and 101 the layers were separated. The aqueous phase was extracted with two 100 m l . portions of ether. The extracts were combined with the ether layer, and the solvent was removed with a rotary evaporator. The product was dried under reduced pressure in the electrical oven ( 4 0 ° C ) and recrysta l l ized from acetone in an ice bath. Y i e l d : 34. 84g. m . p . : 1 5 7 - 1 5 8 ° C . i r (KBr) 3280 (Q=C-N-H), 3020 (=C-H), 1620 c m " 1 (C=0 & C=C); nmr ( C D C 1 3 ) 5" 5. 73 (sharp m , 2, CH=CH-) , 5. 63 - 5. 13 (broad s, 1, N - H ) , 4.1 - 3.53 (m, 1, - C H - C = 0 ) , 5.6 - 0.57 (m, 17, C f e H n & C & H & ) . A n a l . Calcd . for C 1 2 H 2 1 N O : C, 74. 78; H , 1 0 . 1 3 ; N , 6.74. Found: C , 74.29, H , 10.00; N , 6.92. 3. N - c y c l o h e x y l - N - 3-cyclohexenylmethylamine A dry 250 m l . three-necked flask was equipped with a mechanical s t i r rer and Soxhlet apparatus with reflux condenser carrying a drying tube. 200 m l . dry ether was placed in the flask and lithium aluminum hydride (11. 4g, 0.3 mole) was added to it. The solution was refluxed gently with s t i r r ing for 4 hours. . N-cyclohexyl-3-cyclohexene-carboxamide (31. l g , 0. 15 mole) was packed into the Soxhlet extractor with a round-bottomed columnar filter paper to prevent blockage of the siphon a r m . A glass rod was placed in the powder in order to maintain an efficient flow of solvent. . Refluxing was continued so as to maintain slow and steady dissolution of the amide in the Soxhlet. After the 102 addition of amide was completed, the mixture was refluxed for 48 hours with s t i r r ing . The reaction mixture was cooled in an ice-water bath and 30 m l . of distil led water were added dropwise with st i rr ing to decompose excess hydride. When the mixture had turned white, sufficient 40% N a O H was added with rapid st irr ing to give clear separation of the ethereal layer . The ether insoluble layer was separated by centrifugation. The ethereal layer was dried over anhydrous magnesium sulfate overnight. The solvent was removed using a rotary evaporator. The crude product was distilled under reduced pressure and the fraction boiling at 7 5 - 7 8 ° C (0.2 - 0.3 m m . Hg) was collected). Y i e l d : 26.68g (92%). ir (neat) 3300 (weak, NH), 2940,2860 ( C - H 2 ) , 3040 ( = C H - ) , . 1655 c m - 1 (C=C); nmr-(G'DC1 3 ) & 5. 78 - 5. 60 (sharp m , 2, - H C = C H - ) , 2.62 - 2.43 (m, 2, - C H ^ N ) , 2.43 - 0.67 (m, 19, C & H n , C & H ? & N H ) . Mono-hydro chloride derivative The salt was prepared by the same procedure as that described in E x p . A - 3 . A n a l . - C a l c d . for C H N C I : C, 68; H , 10.45; N , 6 .1 ; —~~~— 1 -5 LAQ. C l , 15. 49. • Found: C , 67. 83; H , 10.32; N , 6.23; C l , 15.47. 4. -Chloro-N-3-cyclohexenylmethyl-N-cyclohexylacetamide A 250 m l . three-necked flask was equipped with a mechanical s t i r rer , dropping funnel (125 m l . ) with a drying tube, and side a r m for setting a thermometer (range - 1 0 0 ° - 5 0 ° C ) . N - c y c l o h e x y l - N - 3 - c y c l o -103 hexenylmethylamine (13.51g, 0.07 mole) and dry triethylamine (8. Olg, 0. 08 mole) in 180 m l . dry ether were placed in the flask. The solution was cooled to between -5 and - 1 0 ° C in an ice-sal t bath for one hour, and then a solution of chloroacetylchloride (7. Olg, 0.07 mole) in 40 m l . dry ether was added dropwise to the vigorously s t i rred solution so as to keep the temperature of the reaction below 0 ° C . After the addition was completed, the mixture was st i rred overnight at room temperature. 30 m l . 5% H C l was added to the mixture and it was st i rred for 15 minutes. The layers were separated and the aqueous phase was extracted with two 100 m l . portions of ether. The extracts were combined with the ethereal layer and the solvent was removed using a rotary evaporator. The product was distilled under reduced pressure using a 9 inch Vigreux column warmed with a heating tape. The fraction boiling at 1 5 0 ° C (0. 2 m m . Hg) was collected. Y i e l d : 13.25g(80%). ir (KBr) 3030 (=C-H), 1650 (C=C & C=0), 700 c m " 1 ( C - C l ) ; O nmr (CDC1, ) & 5. 70 (sharp m , 2, - C H = C H - ) , 4 . 15 (s, 2, C - C H - C l ) , ' 2 3.30 - 3.17 (m, 2, - C H ^ N - ) , 2.40 - 0.67 (m, 18, C & H j l & C f e H ? ) . 5. p(--IDimethylamifio-N- 3- cyclohexenylmethyl-N- cyclohexyl- acetamide A 250 m l . three-necked flask was e-quipped with a mechanical s t i r rer and a dropping funnel (125 m l . ) with a drying tube. 150 m l . dry ether was placed in the flask and cooled to between -5 and - 1 0 ° C in an ice-sal t bath for one hour. Dimethylamine (6cc, 0.09 mole) was trapped 104 in a dry graduated cylinder f rom a dimethylamine gas tank using an acetone-dry ice bath, and poured all at once into the reaction flask. A solution of o( -chloro-N -3-cyclohexenylmethyl-N -cyclohexylacetamide ( l l g , 0.04 mole) in 60 m l . dry ether was added dropwise to the reaction mixture so as to keep the temperature of the solution below 0 ° C . After the addition was completed, the reaction was continued at a temperature below 0 ° C for 10 hours and then overnight at a temperature below 1 0 ° C . The dimethylamine salt was suction filtered and the solvent removed using a rotary evaporator. The crude product was distil led under reduced pressure through a 9 inch Vigreux column. The fraction boiling at 1 6 0 ° C (0. 65-0. 75 m m . Hg) was collected. Y i e l d : 10.9g(96%). ir (neat) 3040 (=C-H), 1642 c m " 1 (C=C & C=0); nmr (CDClg) & 5.67 (m, 2, - C H = C H - ) , 3. 5 - VS'OJ^.j l } ( r C H 7 - N h s, 6., N C C H ^ , C J , M ) , 2.18 -0.72(m, 18,(C 6 H u & C f e H )).. , ' ' "' V Methyl iodide derivative 2. 78g O.OJ(mole) of aminoacetamide was dissolved in 1 5 m l . of dry ethanol and 1 Tft/, of methyl iodide was added. The mixture reacted at room temperature. The salt was recrystal l ized from dry ethanol and ether. A n a l . Calcd . for C H N I: C , 51.45; H , 7.85; N, 6.66; I, 30.2. F o u n d : . C, 51.31; H , 7.93; N, 6. 74; I, 30.06. 105 6. N , N - D i m e thy 1 - N 1 - 3- cyclohexenylme t h y l - N ' - cyclohexylethyl-enediamine A dry 250 m l . three-necked flask was equipped with a mechanical s t i r rer and reflux condenser (125 m l . ) with a drying tube. 150 m l . dry ether was placed in the flask and lithium aluminum hydride (2.28g, 0.06 mole) was added to it. . The solution was gently refluxed for 3 hours. A solution of <^-dimethylamino-N-3-cyclohexenylmethyl-N-cyciohexylactamide (8. 34g, 0.3 mole) in 30 m l . dry ether was added dropwise f rom the dropping funnel to the vigorously s t i r red solution so as to maintain gentle refluxing. After the addition was completed, the reaction was continued for 30 hours. Then the flask was cooled to below 0 ° C by an ice-sal t bath and 20 m l . of distilled water was added dropwise to decompose excess hydride. Stirr ing was continued until the mixture turned white in colour. Sufficient 40% sodium hydroxide was added, with st irr ing, to allow clear separation of the ethereal layer . The ether-insoluble material was separated by centrifugation. The ethereal solution was dried over anhydrous magnesium sulfate overnight. The drying agent was suction filtered and the ether was removed using a rotary evaporator. The crude product was distil led under reduced p r e s -sure, and the fraction boiling at 1 1 6 ° C (0. 6 m m . Hg) was collected. Y i e l d : 7. 6g (96%). ir (neat) 3040 (=C-H), 1650 c m " 1 (C=C); nmr ( C D C 1 3 ) '£> 5.66 (m, 2, - C H = C H - ) , 2.20 (s, 6, N - ( C H 3 ) 2 ) , 2. 8 - 0.77 (m, 3 0 ) C 6 H ? , C 6 H 1 1 - C H 2 - N - C H 2 - C H 2 - N ( C H 3 ) 2 ) . 106 Di-perchlorate derivative The salt was prepared by the same procedure as that des-cr ibed in Exp. B-2 , except that two equivalents of perchloric acid was o used, m . p . : 142-143 C . A n a l . Calcd . for C 1 ? C l 2 O g : C , 43.85; H , 7.31; N , 6.02; C l , 15.28. Found: C , 43. 75; H . 7.48; N , 5.87; C l , 15.09. L . N , N - D I M E T H Y L - N ' - 3 - CY C L O H E X E N Y L M E T H Y L - N ' -P H E N Y L E T H Y L E N E D I A M I N E 1. o< - Chloro-N-phenylacetamide 223. 48g (2. 4 m l . ) of aniline and 1 300 m l . dry ether were placed in a 2 litre three-necked flask equipped with a dropping funnel, mechanical s t i r rer , and thermometer (range - 1 0 ° C - 5 0 ° C ) . The solution was placed in an ice-sal t bath for one hour to cool it to below 0 ° C . . Chloroacetyl chloride (1 35. 6g, 1. 2 m l . ) was added dropwise f rom the funnel to the vigorously s t i r red solution so as to keep the tempera-ture of the reaction mixture below - 0 ° C . After the addition was com-pleted, the st irr ing was continued for 3 hours below 0 ° C . and for another 5 hours at room temperature. 200 m l . 5% HC1 solution was then added to the flask and the mixture was st i rred for 10 minutes. The mixture was suction filtered through a Buchner funnel, and a white product was obtained. The anilide was washed in the funnel using dis -tilled water until no HC1 could be detected with litmus paper. The ether layer was dried over magnesium sulfate, and the solvent was flash 107 evaporated to recover the anilide it had dissolved. Y i e l d : 191g (95%). m . p . : 1 3 5 - 1 3 6 ° C . ir (KBr) 3280 ( - C O N H ) , 1670 (C=0), 625 c m " 1 ( C - C l ) ; nmr ( C D C 1 3 ) , & 8.50 - 7.80 (broad s, 1, N - H ) , 7.68.- 6.97 (m, 5, phenyl-H)", 4. 17 (s, 2, C H 2 - C 1 ) . A n a l . C a l c d . for C 0 H„ N C I O: C, 56.6; H , 4. 7; N , 8.2; o o C l , 20.9. Found: C , 56.66; H , 4. 78; N , 8.38; C l , 21.12. 2. oi- Dime thy lamino-N-pheny lac etamide A 2 litre three-necked flask was equipped with a 500 m l . dropping funnel with drying tube, mechanical s t i r rer , and side a r m for setting a thermometer (range - 1 0 0 ° C - 5 0 ° C ) . 250 m l . dry methanol and 750 m l . dry ether were added to the flask. The solution was s t i r red in an ice-sal t bath for one hour until it reached -5 to - 1 0 ° C . D i m e t h y l -amine (132. 4cc, 2 mole) was trapped in a graduated cylinder f rom a dimethylamine gas tank using an acetone-dry ice bath ( - 8 0 ° C ) . It was poured into the three-necked reaction flask al l at once. A solution of -chloro-N-phenylacetamide (169. 5g, 1 mole) in 100 m l . dry ether and 200 m l . dry methanol was added very slowly f r o m the dropping funnel to the vigorously s t i rred solution so as to keep the temperature of the solution below 0 ° C . When the addition had been completed, the mixture was s t i rred continuously at 0 ° C for 10 hours, and overnight at room temperature. The white salt was suction filtered and the solvent was 108 completely removed using a flash evaporator. 30 m l . distil led water was added to the product and it was shaken vigorously in a separatory funnel. The mixture was extracted using two 200 m l . portions of ether, and the ethereal solution was dried overnight with anhydrous magnesium sulfate. The solution was suction filtered and the solvent was removed by flash evaporation. Y i e l d : 152g (85%). b . p . : 1 3 5 ° C (2mm. Hg). ir (neat) 3300 (CONH), 2940, 2825, 2780 ( N ( C H 3 ) 2 ) , 1680 c m " 1 (C=0); nmr (CDC1 3 ) <f 7 . 7 3 - 6.91 (m, 5, phenyl-H), 3 .00(s , 2, 0 = C - C H 2 ) , 2.33 (s, 6, N ( C H 3 ) 2 ) . Mono-hydro chloride derivative The salt was prepared by the same procedure as that described in E x p . A - 3 . m . p . : 1 6 0 . 5 - 1 6 1 ° C . • A n a l . . C a l c d . f o r - C . n H . _ N _ O C l : . C, 55.9; H , 6.9; N , 13; C l , 16.5. Found: C, 56. 05; H , 7.08; N , 12.87;. C l , 16.69. 3. . N , N-dime thy l-N'-phenylethylenediamine A 2 litre three-necked flask was equipped with a dropping funnel (250 m l . ) , reflux condenser with drying tube, and mechanical s t i r r e r . 1 litre dry ether and lithium aluminum hydride (41. 73g, 1. 1 m l . ) were placed in the flask. The solution was gently refluxed with st irr ing for 3 hours. A solution of o(-dimethylamino-N-phenylacetamide (125g, 0. 7 mole) in 150 m l . dry ether was added to the flask f r o m the dropping funnel so as to maintain a gentle reflux. After the addition was com-109 pleted, the mixture was refluxed with st i rr ing for 48 hours. When the reduction was over, the flask was cooled in an ice-sal t bath for 30 minutes and excess lithium aluminum hydride was decomposed with distilled water. Sufficient 40% N a O H solution was added to give good separation from ether insoluble substances. The ethereal layer was dried over anhydrous magnesium sulfate and the solvent removed using a flash evaporator. The product was distil led under reduced pressure on the oil bath. Y i e l d : 109g(95%). b . p . : 8 5 ° C (1 m m . Hg). ir (neat) 3380 (N-H), 2970 - 2775 ( N ( C H 3 ) ) , 1600 c m " 1 (aromatic); nmr ( C D C 1 3 ) & 7.33 - 6. 5 (m, 5, phenyl-H), 4 . 4 7 - 3.92 (broad s, 1, N - H ) , 3.57 - 2.80 (m, 2, N H - C H 2 ) , 2.80 - 2.30 (m, 2, C H 2 - N ( C H 3 ) 2 h 2. 25 (s, 6, N ( C H 3 ) 2 ) . Di-hydrochloride derivative The salt was prepared by the same procedure as that described in Exp . A - 3 . m . p . : 133-133. 5 ° C . • A n a l . Calcd . for C . _ H , _ N_ C l : C, 50.6; H , 7. 5; N , 11.8; — " — — 1U l o Z L C l , 29.9. Found: C, 50. 74; H , 7 . 4 3 ; N , 12.01; C l , 29.71. 4. . N ' - p h e n y l - N ' -( fi - N , N - d i m e t h y l a m i n o ethyl) - 3 - c y c l o - he xe n e c a'r bo xarrii de A 500 m l . three-necked flask was equipped with a mechanical s t i r rer , a side a r m for setting a thermometer ( range -50 - 1 0 0 ° C ) and a dropping funnel (125 m l . ) . . N , N-dimethyl-M'^-phenylethylene-110 diamine ( 19. 71 g, 0. 1Z mole) , redist i l led triethylamine (15. 15g, 0.15 mole) and dry ether ( 300 m l . ) were placed in the flask. The solution was cooled to between -5 and - 1 0 ° C in an ice-sal t bath for one hour with s t i r r ing . A solution of 3-cyclohexenecarboxyl chloride ( 17. 34g, 0. 1Z mole) in 50 m l . dry ether was placed in the dropping funnel, and added very slowly to the vigorously s t i rred solution in the flask so that the temperature of the solution always remained below 0 ° C . After the addition was completed, the reaction was continued at room temperature overnight. . 30 m l . of disti l led water was added to the reaction mixture, which was rapidly s t i rred for 15 minutes. The layers were separated. The aqueous phase was extracted using two 100 m l . portions of ether and the extracts were combined with the ether phase. The ethereal solution was dried over anhydrous mag-nesium sulfate. The drying agent was suction filtered, and the solvent was removed by a rotary evaporator. The residue was distil led under reduced pressure through a 9 inch Vigreux column warmed by a heating tape. The fraction boiling at 1Z0°C (0.3 m m . Hg) was collected. Y i e l d : 27.4Zg (8Z%) . i r (neat) 3050 ( = C - H ) , Z930 - Z750 (N-( C H 3 ) 2) , 1640 c m " 1 (C=C, C=0) ; nmr ( C D C 1 3 ) £ 7 . 5 7 - 7.13 (m, 5, phenyl -H) , 5.63 m , Z, (HC=.CH), 4. 03 - 3. 73 (m, Z, 0 = C - N - C H 2 ) , Z . 6 7 - Z . 3 3 (m, Z, C - C H 2 - N - ( C H 3 ) 2 ) , Z.30 (s, 6, N - ( C H 3 ) 2 ) , 2. Z0 - 1.5 (m, 7, C & H ? ) . I l l 5. N , N - d i m e t h y l - N ' - 3 - c y c l o h e x e n y l m e t h y l - N ' - p h e thy 1 ene diamine r A 250 m l . three-necked flask was equipped with a mechanical s t i r rer , reflux condenser, and dropping funnel. Both the condenser and funnel c a r r i e d drying tubes. L i thium aluminum hydride (5.7g, 0. 15 mole) and 180 m l . dry ether were placed in the flask. The solution was gently refluxed for 3 hours on the heating mantle with s t i r r ing . A solution of N - p h e n y l - N - ( ft-N, N-dimethylaminoethyl)-3-cyclohexenecarboxamide (26. 2g, 0. 095 mole) i n 40 m l . dry ether was placed in the dropping funnel, and added dropwise to the s t i rred solution at a rate that maintained gentle reflux. - After 30 minutes the addition was completed, and the mixture was s t i rred and refluxed for 30 hours. The heating mantle was then replaced by an ice-salt bath, and the solution was cooled below 0 ° C . 30 m l . of distil led water was added dropwise to the vigorously s t i rred mixture in order to decom-pose excess hydride. A sufficient amount of 40% N a O H solution was added, with s t i rr ing, to the mixture to allow for good separation of the ethereal layer f rom the ether-insoluble residue. The ethereal layer was decanted into an Er lenmeyer flask, and dried over anhydrous m a g -nesium sulfate. The drying agent was suction filtered, and the ether removed by a rotary evaporator. The residue was distil led under reduced pressure through a 9 inch Vigreux column warmed by a heating tape. The fraction boiling at 1 6 2 - 1 6 4 ° C (2 m m . Hg) was collected. Y i e l d : 32g (94%). 112 ir (neat) 3040 (phenyl-H) 2980, 2940, 2830, 2780 ( N ( C H 3 ) Z ) , 1650 (C=C), 1600 (aromatic). M . N - C Y C L O H E X Y L - 2 , 5 - C Y C L O H E X A D I E N E C A R B O X A -M I D E 1. 2, 5-cycl6hexadieriecarboxyTic acid (1, 4-dihydrobenzbic acid D r y benzoic acid (20g, 0. 164 mole) was added to 200 m l . anhydrous ethanol in a 2 litre three-necked flask equipped with a mechanical s t i r rer and loose cotton plugs in the side necks. After the benzoic acid had dissolved, 800 m l . of liquid ammonia was passed directly f rom an ammonia tank through dry heavy rubber tubing into the flask, which was cooled by an acetone-dry ice bath. Then 12. 4g (0.54 mole) of sodium was added in small pieces . When about one-third of the sodium had been added, the white sodium salt of the acid p r e c i -pitated, and the mixture foamed furiously. After al l the sodium had been consumed, as evidenced by the disappearance of the blue-black colour, ammonium chloride (29. 2g, 0.54 mole) was added cautiously. The mixture was s t i r red for an additional hour and then allowed to stand until the ammonia had evaporated. The residue was dissolved in 300 m l . water and the solution poured into 300 g of ice and acidified to a pH of about 4 by addition of 150 m l . 10% hydrochloric acid. The resulting mixture was extracted with four 150 m l . portions of ether, and the combined extracts were washed with 50 m l . of a saturated aqueous solution of sodium chloride. The solution was dried over lOg of anhydrous magnesium sulfate. The solution was separated f r o m the 113 drying agent and concentrated at room temperature under reduced pressure by a flash evaporator. The residual oi l was distil led under reduced pressure through a 9 inch Vigreux column. 1, 4 dihydrobenzoic acid was obtained as a colourless o i l . The product was stored under nitrogen in a closed flask. Y i e l d : 15. 6g (78%). b . p . : 9 0 ° C (0.2 m m . Hg). (Li t . b . p . : 8 0 - 9 8 ° C / 0 . 0 1 m m . Hg). i r (neat) 2500 - 3500 ( -COOH) , 1690 (C=0); 1630 c m " 1 (C=C); nmr ( C D C 1 3 ) 6 11. 53 ( - C O O H ) , 6. 20 - 5.62 (m, 4, CH=CH, CH= CH), 4.00 - 3.60 (t, J= 1 0 H 2 , CH-C=0) , 3.00 - 2.55 (m, 2, C 4 H 2 ) . . 2. 2, 5-cyclohexadienecarboxyl chloride 1, 4-dihydrobenzoic acid (14. 9g 10.1 mole) was placed in a 250 m l . three-necked flask. The flask was equipped with a mechanical s t i r rer , reflux condenser and dropping funnel (125 m l . ) . The condenser and funnel both car r ied drying tubes. Thionyl chloride (28. 6g, 0. 24 mole) was added to the acid over a period of five minutes, while the mixture was being rapidly s t i r red . The flask was placed in an oi l bath, and heated to a temperature of 1 5 0 ° C for one hour. The condenser was replaced by a distillation head, 80 m l . of anhydrous benzene was added, and the mixture was disti l led until the temperature of the vapours reached 9 0 ° C . The mixture was cooled and a further 80 m l . of anhydrous benzene added. . The distillation was continued until the temperature of the vapours again reached 9 0 ° C . The cooled residual acid chloride was disti l led under reduced pressure to yield the 2,4-114 cyclohexadienecarboxyl chloride. Y i e l d : 13.5g (79%). b . p . : 9 5 ° -1 0 0 ° C (0. 6 m m . Hg). ir (neat) 3040, 2875, 2820 ( C - H ) , 1770 c m " 1 (C=C, C=0). . 3. . N-cyclohexyl -2 , 5-cyclohexadienecarboxamide A solution of cyclohexylamine (19.84g, 0.2 mole) in 180 m l . dry ether was placed in a 250 m l . three-necked flask. The flask was equipped with a mechanical s t i rrer and a dropping funnel (125 m l . ) which car r ied a drying tube. The solution was cooled to between -5 and - 1 0 ° C in an ice-sal t bath for one hour with s t i r r ing . A solution of 2, 5-cyclohexadienecarboxyl chloride (14. 2g, 0. 1 mole) in 40 m l . dry ether was placed in the dropping funnel, and added dropwise to the rapidly s t i rred solution at such a rate so as to keep the temperature of the solution below 0 ° C . • After the addition was completed, the mixture was st i rred at room temperature for 5 hours. 50 m l . of distil led water was added to the mixture, and it was st i rred for a further 30 minutes. The white product was suction filtered, and the layers separated. The ethereal layer was flash evaporated, and the solid product combined with the suction fil tered one. The product was dried in the air and then under reduced pressure at room temperature. Y i e l d : 16. 3g (79.5%). ir (neat) 3275 (0=C-N-H) , 1630 (C=0, C=C) ; nmr (CDC1 ^ & 6.2. 115 N . N-2 , 6 - D I M E T H Y L P H E N Y L - C Y C L O H E X Y L A M IN E 1. N - 2 , 6-dime thy [phenyl- cyclohexanecarboxamide A 500 m l . three-necked flask was equipped with a mechanical s t i r rer , dropping funnel with a drying tube and thermometer (range - 5 0 ° to 1 0 0 ° C ) . 2, 6-dimethylaniline (48. 27g, 0. 4 mole in 300 m l . ) dry ether was placed in the flask. The solution was cooled to between -5 and - 1 0 ° C in an ice-salt bath for one hour with s t i r r ing . A solution of cyclohexanecarboxyl chloride (29. 123g, 0.2 mole) in 50 m l . dry ether was added dropwise from the dropping funnel to the vigorously s t i rred solution so as to keep the temperature of the solution below 0 ° C . The mixture was then st irred at room temperature overnight. 50 m l . of distilled was added to the flask and the mixture was s t i r red for 20 minutes. The white product was suction filtered using a Buchner funnel and the layers were separated. The ethereal phase was flash evapora-ted to remove the solvent and the solids were combined with the suction filtered product. The product was mixed with 200 m l . 5% HC1 and st irred for one hour on the magnetic s t i r r e r . The solids were again suction fil tered and washed with 500 m l . disti l led water until no trace of HC1 could be detected with litmus paper. The solid was predried in the air and then under reduced pressure in the electr ical oven ( 4 0 ° C , 20 m m . Hg). The product was crystal l ized from a mixture of ether and a small amount of ethylacetate. Y i e l d : 45. 2g (98%). m . p . : 201-2 0 2 ° C . 116 ir (KBr) 3250 ( C O - N H ) , 3040 (phenyl-H), 1645 c m " 1 (C=0); nmr ( C D C 1 3 ) 7. 30 - 6.92 (m, 3, phenyl-H), 2.10 (s, 6, 2, 6-dimethyl), 2.07 - 0.68 (m, 12, C & Hj & NH) . - A n a l . - C a l c d . for C , _ H , . N O : . C , 77.6; H , 9.05; N , 6.04. I D C. 1 Found: C , 77. 82; H , 9.05; N , 6.23. 2. N-2., 6-dimethylphenyl- cyclohexylmethylamine A dry 250 m l . three-necked flask was equipped with a mechanical s t i r rer and Soxhlet apparatus fitted with a reflux condenser carrying a drying tube. 200 m l . dry tetrahydrofuran was placed in the flask and lithium aluminum hydride (7.6g, 0. 2 mole) added to it. The solution was refluxed with s t i r r ing for 3 hours. Then dry cyclohexane-2, 6-dimethylanilide (23g, 0. 1 mole) was packed in the extractor, in which a columnar filter had been placed to prevent blockage of the siphon a r m by the amide. A glass rod was inser ted in the powder to allow efficient flow of the solvent. The rate of reflux was controlled so that the amide would keep dissolving slowly. The mixture was then refluxed for 6 days until no unreduced amide could be detected. The flask was cooled below 0 ° C in an ice-sal t bath. 50 m l . of disti l led water was added slowly to the mixture to decompose excess hydride. The st i rr ing was continued until the mixture became white in colour. Sufficient 40% sodium hydroxide solution was added to separate the ethereal layer f rom the ether-insoluble residue. The residue was separated by centrifugation and the ethereal layer was dried over 117 anhydrous magnesium sulfate overnight. The solvent was removed using a rotary evaporator and the crude product was distil led under reduced pressure . Y i e l d : 20.6g(95%). b . p . : 1 2 2 ° C / 0 . 5 m m . Hg. ir (neat) 3380 (NH), 3050 (aryl-H) , 1600 c m " 1 (aromatic); nmr ( C D C 1 3 ) i 7.11 - 6.57 (m, 3, phenyl-H), 2.91 (s. 1, NH), 2 . 7 5 ( m , 2, C H - C H 2 ) , 2.23 (s, 6, dimethyl), 2.00 - 0.80 (m, 11, C 6 H l l ) ' G . N , N - D I M E T H Y L - N ' - C Y C L O H E X Y L M E T H Y L E T H Y L E N E -D I A M I N E 1. o£ -dimethylamihoethyl-N-cyclohexanecarboxamide A 250 m l . three-necked flask was equipped with a mechanical s t i r rer , dropping funnel (125 m l . ) with a drying tube, and side a r m for setting a thermometer (range -100 to 5 0 ° C ) . N , N-dimethylethyl-enediamine (26.45g, 0.3 mole) in 170 m l . dry ether was placed in the flask. The solution was cooled to between -5 and - 1 0 ° C in an ice-sal t bath for one hour. A solution of cyclohexanecarboxyl chloride (21.85g, 0. 15 mole) in 30 m l . dry ether was added very slowly f rom the funnel to the rapidly s t i r red solution so as to keep the temperature of the solution below 0 ° C . After the addition was completed, the st irr ing was continued at 0 ° C for one hour and at room temperature overnight. 30 m l . distil led water was added and the mixture was st i rred for 15 minutes. . The solid was suction filtered and the layers were separated. The aqueous phase was extracted with two 100 m l . portions of ether. 118 The extracts were combined with the ether layer and dried with 50g of anhydrous magnesium sulfate. The drying agent was suction filtered and most of the solvent removed using a rotary evaporator. The crude product was recrysta l l ized from a mixture of petroleum ether ( 3 0 ° - 6 0 ° C ) and ethylacetate. Y i e l d : 28. l g (95%). m . p . : 8 6 ° C . i r (neat) 3280 (CONH), 2920, 2850, 2780 ( N C H ^ ) , 1635 c m " 1 (C=0); nmr ( C D C 1 3 ) r> 3. 47 - 3. 10 (m, 2, N H - C H 2 ) , 2. 50 - 2. 22 (s, 6, N ( C H 3 ) 2 ) , 2.00 - 0.40 (m, 12, C & H & N H ) . - A n a l . Calcd . for C H N O: C, 66.65; H , 11. 1; N , 14. 15. " 11 Z Z Found: C, 66.79; H , 11.05; N , 14.06. 2. . N , N-dimethyl-N'-cyclohexylmethylethylenediamine A 250 m l . dry three-necked flask was equipped with a mechanical s t i r rer , dropping funnel (120 m l . ) and reflux condenser. The funnel and condenser both carr ied drying tubes. 180 m l . dry ether was placed in the flask and lithium aluminum hydride (9.88g, 0. 26 mole) was added. The solution was gently refluxed for 3 hours with s t i r r ing . A solution of d. -dimethylaminoethyl-N-cyclohexane-carboxamide (25. 65g, 0.13 mole) in 30 m l . dry ether was added dropwise to the hydride solution f rom the dropping funnel so as to maintain a gentle reflux. After the addition was completed, the mixture was refluxed for 36 hours. When the reduction was over, the heating mantle was replaced by an ice-water bath and 20 m l . distil led 119 water was added slowly to the vigorously s t i rred mixture in the flask to decompose excess hydride. . St irr ing was continued until the mixture became white. Sufficient 40% N a O H was added to allow clear separation of the ethereal layer f rom the ether-insoluble residue. The residue was separated by centrifugation. The ethereal layer was dried over anhydrous sodium sulfate overnight. The solvent was removed using a rotary evaporator, and the product was distilled under reduced pressure through a 9 inch Vigreux column. . Y i e l d : 22g(96.5%). b . p . : 7 3 ° C (2. 0 m m . Hg); 6 6 ° C (0. 5 m m . Hg). ir (neat) 3340 (NH), 2920, 2760, 2680 ( N ( C H 3 ) 2 ) ; n m r ( C D C l 3V I 2 . 7 - 2 . 2 3 (m, 6, C H ^ N - C H 2 - C H )', 2.10 (s, 6, N ( C H ) ), 2.03 - 0.6 (m, 12, C & H u & NH). Di-perchlorate derivatives The salt was prepared by the same procedure as that des-cribed in E x p . B -2 , except that two equivalents of perchloric acid was used. m . p . : 1 8 3 - 1 8 4 ° C . A n a l . Calcd . for C H N O C l : C, 34.09; H , 6. 76; 11 26 2 8 2 N , 7.28; C l , 18.47. Found: C, 34. 13;. H , 6.81; N , 7.13; C l , 18.63. P . . S I N G L E R E A C T I O N S 1. 3 - chloro-4 -cyc lohexene- l , 2-dicorboxylic acid anhydride 1-chlorobutadiene (17g, 0.2 m l . ) , maleic anhydride (19g, 0. 2 ml.-), iodine (340 mg) and a small amount of hydroquinone with 120 ZOO m l . benzene were placed in a Z50 m l . flat-bottomed flask equipped with a reflux condenser. The reaction mixture was refluxed while being s t i rred on an oi l bath for 48 hours. Then the iodine was removed with sodium hypochlrite. The precipitated crystal was separated to yield 26g of the product. It was then recrysta l l ized from a mixture of benzene and acetone, m . p . : 1 3 0 - 1 3 1 ° C . i r (KBr) 3040 (=C-H), 1860, 1830, 1780 ( - C O - 0 - C O - ) , 1650 c m - 1 (C=C); nmr (C$C13) & 6.17 - 6.1 (m, 2, CH=CH), 5.08 -4.77 (m, 1, C H - C 1 ) , 3.6 - 3.27 (m, 2, 0 = C - C H - C H - C = 0 ) , 2. 9 - 2.58 (m, 2, C = C - C H 2 - C ) . A n a l . Calcd . for C g H ? C I O 3 : . C , 51.5; H , 3.76; C l , 19.02. F o u n d : . C, 51.63; H , 3.83; C l , 18.88. 2. . Cyclohexyl p - to lime sulfonate A 500 m l . three-necked flask was equipped with a mechanical s t i r rer and thermometer (range - 5 0 ° C to 1 0 0 ° C ) . Cyclohexanol (50g, 0. 5 mole) and pyridine (158g, 2 mole); were placed in the flask. The flask was i m m e r s e d in an ice-sal t bath to lower its temperature below 1 0 ° C . Then powdered p-tolunesulphonyl chloride (105g, 0.55 mole) was added in portions over a period of 30 minutes so that the temperature did not exceed 1 0 ° C . The mixture was then s t i r red for 5 hours below 1 0 ° C , after which it was diluted with 300 m l . of hydrochloric acid (SP gr . 1.19, 38%) in 1 litre of ice water. The crystal l ized ester was collected on a 121 chilled Buchner funnel and allowed to dry in the a i r . . It was then dried under reduced pressure at room temperature. The ester was r e -crystal l ized from petroleum ether ( 3 0 - 6 0 ° C ) . Y i e l d : 101. 7g-(80%). m . p . : 4 4 - 4 5 ° C . i r (KBr) 3040 (phenyl-H), 1600 (aromatic) 1180 ( - S C ^ - O ) ; nmr ( C D C 1 3 ) 0 7.95 - 7.2 (m, 4), 4. 8 - 4. 3 (m, 1, O - C H ) , 2.4 (s, 3, p - C H 3 ) , 2.33 - 0.93 (m, 10, C 6 H 1 Q ) . A n a l . C a l c d . for C 1 3 H l 8 S 0 3 : C , 61.45; H , 7.08; S, 12.6. Found: C, 61.31; H , 7. 18;. S, 12.44. 3. 1-Chloro 3-cyclohexene A dry 500 m l . three-necked flask was equipped with a mechanical s t i r rer , reflux condenser with a drying tube, and liebig condenser connected to a nitrogen inlet. 3-cyclohexenecarboxylic acid (63. 3g, 0. 5 mole), lead tetracetate (lOOg, 0. 2258 mole) and 300 m l . dry benzene were placed in the flask under nitrogen gas. The powdered lithium chloride (9. 55g, 0.2258 mole) was added to the solution and r e -fluxed while being s t i rred until the powder disappeared and no carbon dioxide was evolved. The solution became light yellow. Two 200 m l . portions of distil led water were added with rapid st irr ing and the benzene layer was removed with a separatory funnel. The benzene solution was dried by anhydrous magnesium sulfate. The drying agent was suction filtered and the solvent was removed using a rotary eva-122 porator. The crude product was distilled under reduced pressure and the fraction boiling at 1 4 0 - 1 4 1 ° C was collected. Y i e l d : 21g (80%). ir (neat) 3040 (=C-H), 2940, 2910, 2830 ( C H 2 ) , 1645 (C=C), 670 ( C - C l ) ; nmr (CDC 13) 6 5. 67 - 5. 3 (m, 2, CH=CH), 4. 37 - 3. 87 (m. 1, > C H - C 1 ) , 2.87 - 1.55 (m, 6, C 6 H & ) . A n a l . Calcd . for C 6 H ? C l j ! C, 61.90; H , 7.74;. C l , 30. 06. Found: C , 61.95; H , 7 . 9 1 ; C 1 , 30.23. 4. 3- Cyclohexenylmethylamine A 250 m l . three-necked flask was equipped with a dropping funnel (125 ml. ' ) , reflux condenser with drying tubes, and mechanical s t i r r e r . 150 m l . dry ether and L A H (5. 7g, 0. 15 mole) were placed in the fla.sk-. The solution was gently refluxed while being s t i r red for 3 hours. A solution of 3-cyclohexene nitri le (10. 7g, 0. 1 mole) in 40 m l . ether was added to the flask from the dropping funnel so as to maintain a gentle reflux. After the addition was completed, the mixture was refluxed while being s t i r red for 40 hours. . Then the flask was cooled in an ice-sal t bath for 30 minutes, and excess lithium aluminum hydride was decomposed with distil led water. Sufficient 40% N a O H was added to give good separation from the ether insoluble residue. The ethereal layer was dried over anhydrous magnesium sulfate. The solution was suction filtered through a Buchner funnel and the ether was removed using a rotary evaporator. The product was dist i l led. Y i e l d : 10.29g (70%). b . p . : 1 6 9 - 1 7 0 ° C . 123 i r (neat) 3360, 3280 ( N - H 2 ) , 1650 c m " 1 (C=C); nm4 ( C D C 1 3 ) & 5 . 9 3 - 5.5 (m, 2, CH=CH), 2. 8 - 2. 5 (d, 2, J= 5, C H - N ) , 2 . 5 0 -0. 83 (m, 9, C 6 H 7 k N H 2 ) . Mono-hydro chloride derivative The salt was prepared by the same procedure as that described in E x p . A - 3 . m . p . : 2 1 6 ° C . A n a l . Calcd . for C ? H 1 4 N C 1 : C, 56.9; H , 9. 5;. N . 9.5; C l , 24.07. Found: C, 56.90; H , 9.61; N , 9.49; C l , 24.01. 5. oj - C h l o r o - N , N-dirhethylacetamide A 250 m l . three-necked flask was equipped with a mechanical s t i r rer and a dropping funnel with a drying tube. 140 m l . dry ether \was placed in the flask and cooled by an acetone-dry ice bath for 30 minutes. 26.4 m l . of dimethylamine (0.4 mole) was added to the flask. • A 60 m l . solution of chloroacetyl chloride (22. 6g, 0. 2 mole) in dry * ether was added very slowly f r o m the dropping funnel so as to control vigorous reaction. After the addition was completed, the reaction con-tinued at the same temperature for 3 hours . Then the mixture was s t i r red at room temperature for 2 hours. The white dimethylamine hydro chloride was filtered through a Buchner funnel. The solvent was flash evaporated, and the product was disti l led under reduced pressure . Y i e l d : I4.52g (60%). b . p . : 6 3 - 6 4 ° C (1.1 m m . Hg). i r (neat) 2920 ( N ( C H 3 ) 2 ) , 1650 (C=Q) 790 c m " 1 ( C - C l ) ; nmr 124 ( C D C 1 3 ) <S 4.13 (s, 2, C 1 - C H 2 ) , 3.05 (d, 6, J= 7 Hz, N f C H ^ ) . 6. o£"- CKloro - N - 2, 6 - dime thy Ipheny 1 - N - cy clohexy Ime thyl -acetamide (attempted) A 250 m l . three-necked flask was equipped with a mechanical s t i r rer , dropping funnel (125 m l . ) with drying tube, and a side a r m for setting a thermometer (range -50 to 1 0 0 ° C ) . N - c y clohexy Ime thyl-2, 5-dimethylamine (6.5g, 0.03 mole) and dry triethylamine (4. 04g, 0.04 mole) in 150 m l . dry ether were placed in the flask. The solution was cooled to - 8 0 ° C in an acetone-dry ice bath for one hour. A solution of oc" -chloroacetyl chloride (3. 642g, 0. 03 mole) in 40 m l . dry ether was placed in a dropping funnel and added dropwise to the vigorously s t i rred cold solution. . The product appeared to have been formed, but at the end of the reaction, the mixture started to darken from a light brown colour. At room temperature, it was impossible to isolate the desired product due to its further instability. 7. <xC - Chloro-N-diphenylacetamide (attempted) A 250 m l . three-necked flask was equipped with a mechanical s t i r rer , dropping funnel (125 ml.) with drying tube, and a side a r m for setting a thermometer (range -50 to 1 0 0 ° C ) . Diphenylamine (8.45g, 0.05 mole) and dry triethylamine (5. 05g, 0.05 mole), in dry ether (150 m l . ) , were placed in the flask. The solution was cooled in an acetone dry ice bath ( - 8 0 ° C ) for one hour. A solution of °i - c h l o r o -acetylchloride (5. 65g, 0.05 mole) in dry ether (30 mole) was placed in 125 the dropping funnel, and added dropwise to the rapidly s t i rred solution over a period of 20 minutes. The st irr ing was continued in the acetone-dry ice bath for 5 hours at room temperature. The triethylamine salt was suction filtered, and the ether removed by a rotary evaporator, but the product rapidly decomposed into a dark brown tarry substance. P A R T V 126 H I S T A M I N E The ubiquitous distribution of histamine in plants, lower animals and mammals , and the variety and significance of its biolo-gical effects have interested physiologists, pharmacologists and chemists for a long time (16). jb - (4- imidazolyl) ethylamine (histamine) was synthesized as a chemical curiosity in 1907 by Windaus and Vogot, before its detection as a uterine stimulant and intense depressor and its isolation in pure form from extracts of ergot in 1910 (Barger and Dale, 17). Histamine research has expanded in many directions since 1910. Methods of synthesis of histamine and its analogues, estimation of histamine in tissues and body fluids, histochemical aspects of E h r l i c h ' s mast cel l , association of histamine and heparine in cells and tissues, and the relationship between structure and activity of analogues of histamine, have all been investigated. . The pharmacology of histamine, in particular its absorption and toxicity; its actions upon smooth muscle , the cardiovascular system and on salivary, brain and gastric secretion; and its relationship with the autonomic nervous system have been investigated. The problem of histamine release by venoms and toxins, physical and chemical agents, basic compounds, and macromolecular releasers like anaphylatoxin, dextran, egg white, etc. in anaphylaxis has also been studied. The mechanism of histamine 127 release and its relationship with respiration, carbohydrate metabolism and phosphorylation has been discussed in connection with the practical problems of anaphylaxis and allergy. The metabolism of histamine, its catabolism and excretion in vivo, its metabolism in vitro (exclusive of diamine oxidase), the enzymatic formation of histamine f r o m histidine, pyridine coenzyme interactions, the metabolism of histamine in man and its distribution in the brain have been investigated. The physiological significance of histamine, including the role of histamine in vasodilation; the effect of histamine on gastric secretion; its role in inflammation, allergy and experimental asthma were other challenges that have been investigated in recent decades. Little is known about the physiological role of histamine in humans. Its function in most of the pathological conditions in which it has been studied also remains uncertain. In the late fifties, Schayer and Kahlson independently showed that there was a great difference in the rate of endogenous., histamine formation under, normal and patholo-gical conditions, without any corresponding changes in tissue histamine content. Two methods for determination of histamine give consistently reproducible results : the whole body histamine formation m a y b e deduced from the urinary excretion of non-isotopic histamine and its metabolites; or one of several adaptations of Schayer's isotopic method may be used. The actions of endogenously formed histamine cannot be reproduced by injected extracellular histamine. Kahlson discovered 128 the high "histamine forming capacity" of embryonic and wound tissues. Histamine forming capacity (HFC) refers to the rate at which histamine is formed in minced tissues, cell suspensions, or the whole body; it denotes histidine decarboxylase activity in numerical terms (11). Kahlson also proposed a functional difference between mast cell and non-mast cell histamine. When methods of determining the rate of histamine formation had been developed, and it was discovered that marked alteration in histidine decarboxylase activity occurs under par t i -cular conditions, new terms were introduced to distinguish histamine liberated from mast cells f rom histamine formed by histidine decar-boxylase activity. Newly formed histamine, which results f rom elevated histidine decarboxylase activity and is believed to act at various sites in the body or in the cell where it is formed, is referred to as "induced histamine" by Schayer (13) and "nascent histamine" by Kahlson (14). The major source of stored histamine in mammalian tissue is the mast cell (15) even though there are other, and possibly more important tissue sources of histamine f rom the standpoint of normal physiology (18). Recognition of the mast cell as a source of histamine in blood and tissue has led to a better understanding of the location and binding of histamine, and the mechanism of its liberation by chemical substances and anaphylaxis in some species. There are other areas of the body such as the epidermis which contain histamine not associated with mast cells . In the stomach of the rat, histamine is found in cells 129 in the superficial layer of mucosa which resemble mast cel ls , in typical mast cells in the submucosa and muscular layers , and in enterochromaffin-like cells in the mucosa (25). Much of the histamine in peripheral nerves is found in the mast c e l l s . . The post-ganglionic fibers of the sympathetic nervous system are extraordinarily r i c h in histamine (27, 28). The majority of histamine in the brain is associated with the phylogenetically old portions of the brain , which also contain large amounts of serotonin and nor-epinephrine. Some of the histamine associated with parts of the bra in such as the postrema, choroid plexus, pineal body, and posterior pituitary gland is in the mast cells (29). The histamine found in mast cel ls , basophils, and the brain, is held in subcellular part ic les . . The particular characteristics of mast cells depend on the presence of heparin in them. In rat mast cells , histamine is stored in basophil granules consisting mainly (over 95% of the granule dry weight) of a heparin-protein complex; heparin is a polymer of glucuronic acid and glucosamine containing sulfate groups linked to some of the hydroxy and amino groups of the sugars. In the blood of most species, all or most of the histamine is found in the basophillic leucocytes (26), which sufficiently resemble mast cells for them to be re fer red to as blood mast cells , just as mast cells have been called tissue basophils. The matrix of the mast ceil granules consists of an insoluble heparin-protein complex held together by strong attractive 130 forces between oppositely charged acid heparin and basic protein macromolecules . The sulphuric acid groups of the heparin were COO „ ^ H e p ( ^ N H S 0 3 ^ H 3 N — - ^ R - C O O H ^ C H ^ - C H ^ - C - N ^ ^ 'OSO3 H , ! ^ " " " ^ C H - N H Figure 8 . Hypothetical heparin-protein-histamine complex of mast cell granules at pH7 thought to be essential in forming the heparin-histamine complex since desulphated heparin lacks the ability to form the complex (20); but it is questionable whether granular heparin present as part of the heparin-protein complex has any sulphuric acid groups available for histamine binding, since these groups are probably involved in heparin-protein linkage. Recent observations indicate carboxyl groups, but not sulphuric acid groups,are the most likely ionic binding sites for histamine in the mast cell granules (21). Quantitative observations on the formation of a complex of heparin-protein with sodium, histamine and other biogenic amines, as well as titration studies, yielded data on the nature, number and location of ionic binding sites on the heparin-protein-histamine complex (?24). If histamine was stored in the granules in ionic linkage, the storage capacity should vary with the degree of dissociation. The influence of pH was studied on granules isolated from 131 14 mast cells containing C . .-histamine which had been endogenously formed during incubation of mast cells in a medium containing 14 C : -his t idine . The granules were isolated by hydrolys is of the cells, and the stores of endogenously formed histamine showed the same pH dependence as that observed for exogenous? histamine (ZZ, Z3). At pH 7 histamine is monobasic, but the ionization increases rapidly below neutrality and it becomes dibasic at pH 3-4. Consequently, the affinity of histamine for cationic binding sites wil l increase with falling p H . On the other hand, the ionization of carboxyl groups in the granules wil l be reduced with increasing acidity, thereby diminishing the histamine-binding capacity of the granules. Therefore , histamine uptake of the granules should be expected to show a pH dependence curve with two inversely changing dissociation curves reflecting a falling dissociation of carboxyl groups and increasing dissociation of amino groups (Z4). The assumption that mast cell histamine is stored in ionic linkage with C O O groups and can be exchanged with other cations (e.g. sodium ions) means that the mast cell granules should have the properties of a weak cation exchange material with carboxyl groups as maximal binding sites (24). There are many substances that release histamine from tissues (25, Z6). Those that release histamine from the mast cell can be placed in two groups. Agents of one group require oxygen to induce histamine release in cat and rat tissues. An example of a compound 132 belonging to this group is "compound 48/80','a condensation product of p-methoxy-phenethyl methyl amine and formaldehyde. Histamine released by these agents is blocked by anoxia and metabolic glucose. The other group; is composed of organic bases such as morphine and aliphatic bases like octylamine which cause histamine release under anoxic conditions, presumably by displacing histamine f rom its binding in the mast cell granules. Uvnas proposed that histamine release is in principle a two-stage process . The first event is degranulation of the mast cell which results in exposure of the histamine-containing granules to the extra-cellular f luid. The second event is the release of histamine from these granules by a simple cation exchange between granular histamine and extracellular sodium (Fig . 9 and ref. 32). E lec t ron microscopic observation may help to clarify the relationship between degranulation and histamine release (33, 34). The exocytosed granules of the mast cells become exposed to the extracellular fluid and release their histamine and 5-hydroxytryptamine. Other granules remain inside the mast cell (Fig , 10 ). About 90 years after the discovery of mast cells , they were clearly differentiated f rom other varieties of connective tissue cells , but no function for them has yet been demonstrated. No evidence has been produced to show that histamine is released from mast cells under physiological conditions. The discovery of the presence of large amounts 133 Releaser attaches 2. to mast cell membrane Granules are discharged Histamine is released f rom discharged granules by cation exchange Heparin -j _ + Ptcuin V COO-HI Complex 1 + • Ha Figure 9 . Degranulation and histamine release in mast cel ls : suggested mechanism of action of compound 48/80 (ref. 3G> ). Figure 10. Schematic drawing to show the sequential exocytosis of histamine-containing granules (ref. ). 134 of histamine in the mast cel l , the high mast cell content of many-tissues, and the finding that virtually all the blood histamine is present within the basophils or "blood mast c e l l s " leads to the belief that the majority of the body histamine resides in these cells 036). The exact proportion of the total body histamine contained in mast cells has not been established. . During histamine release in anaphylaxis, the excretion and catabolism of histamine do not keep pace with its release. There is no evidence that the physiological importance of the mast cells is connected with their content of histamine. The role of histamine and heparin in the mammalian mast cell would appear to warrant further investigation. . Schayer discovered that mast cells collected f r o m the • 14-rat peritoneal cavity formed histamine from C -hist idine. He inferred that the histamine formed by rat mast cells was bound in a stable condition and that exogenous:': histamine was not bound. Although the mast cells contain a lot of histamine, presumably owing to the long l i fe -time of the amine formed in them, their histamine forming capacity is not part icularly high compared with some other tissues. In the skin, which is r i c h in mast cells , the histamine forming capacity is low. In fact, the H F C is so low that on removing the histamine f r o m the mast cells of the skin, it takes 2-3 weeks to restore the original content of histamine. The mast cell is not unique in that the histamine formed in it has a long l i fe . Non-mast cells exist in rat lungs and intestines 135 15 14-U;<Uae\eaeet:c acid l.4-M«tHy|iimda*o»« *.e«U\4hyd« Figure 11. Th« catlbol'ism of histemiive. 136 which resemble mast cells in their complement of long-lived histamine. These non-mast cells differ from real mast cells in that their histamine is resistant to mobilization by compound 48/80. A l s o , the histamine of intestine cells is not released in anaphylaxis. There are considerable differences between species r e -garding the catabolism of histamine both in vitro (36) and in vivo (37, 38). These differences have also been found in studies of histamine formation. Figure 11 shows the main pathways for the catabolism of histamine. A n important pathway in most species, including man, is direct oxidative deamination to imidazoleacetaldehyde (2 in F i g . 11 ) by diamine oxidase, or histaminase which may not be the same as diamine oxidase (39, 40, 41). This pathway in vivo is blocked by diamine oxidase inhibitors. The end products of the reactions catalyzed by diamine oxidase and monoamine oxidase are the same (52, 53). However, monoamine oxidase does not seem to be important in the first step of histamine catabolism in vivo (54). The first evidence for the participation of diamine oxidase in the catabolism of physiological quantities of histamine in vivo was seen in the rat (42). In vitro inhi -bitors of diamine oxidase which do not inhibit monoamine oxidase, isoniazid (43) and aminoguanidine (44), prevent the formation of the major metabolite 1-ribosylimidazole acetic acid, as observed by Kar ja la _et al.(45) and independently by Tabor and Hayaishi (46). The test for diamine oxidase activity required aminoguanidine to be used in 137 a dose that nearly caused complete inhibition in vivo (56), and the 14 intestinal C -histamine level was markedly increased. Diamine oxidase probably dominates intestinal histamine catabolism. A s imilar conclusion was drawn from in vitro studies (57). Since aminoguanidine 14 also reduces total intestinal C -histamine, it is conceivable that the newly-formed endogenous histamine accumulates, partially f i l ls the binding sites, and reduces the uptake of injected C 1 ^ -histamine; or that aminoguanidine may occupy histamine binding sites in the intestine 14 and reduce uptake of injected C -histamine. Almost all the imidazole-acetaldehyde - (2 in F i g . 11 ) i:s converted to imidazoleacetic acid (3 in F i g . 11 ) by aldehyde dehydrogenase. Only a small portion of the aldehyde is reduced by alcohol dehydrogenase to histidol (imidazole-ethanol (4 in F i g . 11. , 47)). Some of the imidazoleacetic acid is con-verted to ribotide (5'-phosphoribosylimidazoleacetic acid (5 in F i g . 11 )). The formation of this ribotide requires 5 ' -phosphoribosyl-1-pyrophos-phate and stoichiometric amounts of adenosine triphosphate (48). The most important pathway for histamine metabolism comprises methylation of the ring nitrogen to l -methyl-4( ^ -amino- / ethyl)imidazole (7 in F i g . 11 ) by histamine methyl transferase. The product of this reaction is usually called methylhistamine or sometimes 1, 4-methylhistamine (7 in F i g . 11 ) to differentiate it f rom the 1 ,5 - isomer (8 in F i g . 11). The imidazole N-methyltransferase acts on no substrate except histamine so far as is known (49, 50). S-adenosylmethionine is the methyl donor. Methylimidazole (7 in 138 F i g . 11 ) is oxidatively deaminated to form the corresponding methyl-imidazole acetaldehyde (9 in F i g . 11 ) in vivo largely by a monoamine oxidase or monoamine oxidase-like enzyme but not by a diamine oxidase (51). In mice l iver , the monoamine oxidase inhibitor amino-guanidine has little effect. These facts suggest that monoamine oxidase, not diamine oxidase, is the catalyst in the formation of methylhistamine (55). Methylhistamine is the in vivo inhibitor of the histamine methylating enzyme that has been known longest. The inhibition of histamine methylation by methylhistamine made it possible to use urinary histamine levels as cr i ter ia of histamine formation and release. By the action of aldehyde dehydrogenase, methylimidazol -acetic acid (11 in F i g . 11) is formed, and this is the major urinary product of histamine catabolism. Therefore, the two major routes, of catabolism are methylation and oxidation. Quantitative analysis of the 14 urine of animals injected with C i . -histamine show that acetylation of the amino group of the side chain is a minor reaction in mammalian tissues (58). Intestinal bacteria acetylate histamine (59) and acetyl-histamine may be present in the urine of some species in considerable amounts (60). N-acetylhistamine (13 in F i g . 11 ) has been shown to be identical with the conjugated histamine that becomes physiologically active after acid hydrolysis (60). Methylation of the p r i m a r y amino group ( la in F i g . 11 ) and the nitrogen of the tautomer (lb) yields 139 N-methylhistamine (14) and 1, 5-methylhistamine (8 in F i g . 11 ), r e s -pectively. Other catabolic pathways are described in ref. 61. Injected histamine cannot reveal precisely what is happening to endogenously formed histamine, because injected histamine is cata-bolized p r i m a r i l y by those organs with the greatest mass and blood flow, like the kidney and l iver . The actions of endogenously formed histamine cannot be reproduced by injected extracellular histamine. This applies particularly to nascent histamine, the metabolic action of which appears to be linked to the very process of its formation. Again, it should be emphasized that the indications are that endogenously formed histamine is catabolized in a different way from injected histamine. This circumstance should encourage more extensive development of non-isotopic methods of determining the principal histamine catabolites. Many cells can take up histamine that is ingested or formed from histidine by the bacterial f lora . . However, almost al l the histamine that is found in tissues is formed in the cells by decarboxylation of histidine, a process which requires pyridoxal-5-phosphate (62, 63). Histidine decar-boxylating enzyme (L-histidine carboxylase) was first discovered i n m a m m a -lian tissue by Werdle. The enzyme is now known to be identical with DOPA-decarboxylase (3, 4-dihydroxy-L-phenylalanine-decarboxylase) . M o r e recently, Schayer has found that at least two enzymes capable of decarboxylating histidine occur in mammals (74). One is a nonspecific 140 N C H 2 - C H - N H 2 C O O H - c o 2 C H 2 - C H 2 - N H 2 A H Histidine Histamine Figure 12. Decarboxylation of histidine L - a m i n o acid decarboxylase, and the other is a specific histidine decarboxylase. These two enzymes are distinguished by their substrate affinities, different pH optima, and their sensitivities to different inhibitors (75) . The specific decarboxylase which is found in the stomach, mast cells , and fetal, l iver of the rat acts only on histidine, which is responsible for induced or nascent histamine. Another enzyme, named aromatic L -amino acid decarboxylase (76), can also decarboxylate histidine in vitro. It has a low affinity for histidine and is closely related to, or identical with, DOPA-decarboxylase . The enzyme or enzymes with low affinity for histidine are called "non-specif ic" histidine decarboxylase. Kahlson, though, is against referr ing to enzymes with a low affinity for histidine as histidine decarboxylase, even with the prefix "non-spec i f i c " . In rats fed on a partly synthetic histamine-free diet without pyridoxal-5-phosphate, the whole body H F C can be reduced to approximately 50% within 2 weeks and to 20% in 6-7 weeks (1£). Semicarbazide is* shown to inhibit histamine formation in vivo, but when it is given to rats in conjunction with a 141 pyridoxine-deficient diet, its effect is very potent (12). Inhibition of histamine formation in vivo was first reported by Schayer et a l . who found that injection of cortisone inhibits histamine formation in the skin but not in the stomach of rats . It was also demonstrated that prednisolone greatly reduces the histidine decarboxylase activity of part icle-free extracts of rat lung (64), but these steroids are relatively weak inhibitors (<12). ^» -3 ,4 -dihydroxyphenyl- Ok. -methylalanine ( Ok - m e t h y l - D O P A ) in vitro strongly inhibits histamine formation in kidney of the guinea pig and rabbit. By contrast, E h r l i c h ascites tumor ( 70) is not inhibited by £><. -methyl -D O P A . Neither does significant reduction of the whole-body H F C occur in rats after they are given toxic dosages of £K -methyl D O P A subcutaneously ( 12 , 69). 4-Bromo-3-hydroxybenzyloxyamine was recognized as an inhibitor of histidine decarboxylase in vitro ( 70) and in vivo ( TV>, 76). Hydrazine derivatives of carbonyl reagents, which engage pyridoxal phosphate, are also known to inhibit amino acid de-carboxylase in vitro (73). In rats, in vivo isonicotinic hydrazide even in high daily doses over a period of two weeks caused only a slight reduction in whole-body H F C , and even this reduction subsided during the course of injections (1S). Ok - H H , the CK -hydrazino analogue of histidine, is reported to be an inhibitor of histidine decarboxylase both in vitro and in vivo ( 78). Weissbach et a l . found that in vitro c( -methylhistidine inactivated the histidine decarboxylase of both mouse mast cells and guinea pig kidney. It was later shown that histidine decarboxylase converts these inhibitors to c<* -methylhistamine 142 which is likely to stimulate histamine, thereby concealing any inhibition that might have taken place. Kahlson et a l . have demonstrated that -methylhistidine is a powerful inhibitor of histidine decarboxylase in vitro and in vivo and have come up with evidence suggesting that the mode of action of ,^-methylhistidine is by competition with the substrate (<1£). o(.-methylhistidine has been shown to be a specific and nontoxic inhibitor of histamine formation in vitro and in vivo using both non-isotopic and isotopic methods. ^ -methylhis t idine is a safe means for identifying the presence of histidine decarboxylase activity and distinguishing this activity f rom that of D O P A decarboxylase (7*3). In work oniinhrbition of histamine formation, the distinction between mast cell and non-mast cell histamine must again be made. • Schayer, and Kahlson et a l . showed that a consistent relationship between tis sue histamine content and histamine forming capacity (HFC) of tissues does not exist. A n initial lowering of tissue histamine content presumably elevates tissue H F C by a feed-back relationshipbetween his tamine content and histidine decarboxylase activity, a repression which operates in vivo but not in vitro. Accordingly, the degree of inhibition manifested in vivo would represent the balance resulting from inhibited and newly formed enzyme molecules . In the inhibition study (-12), a feed-back relationship was f i rmly established as operating in the gastric mucosa (77, 78). Information concerning turnover rate (lifetime of intra-cellular histamine) has been obtained by two different approaches. Schayer injected C ^ - h i s t i d i n e into guinea pigs and rats and determined the amounts of C^^-his tamine present in excised tissues at various 143 time intervals (1 $5,186 )# Kahlson et a l . made use of the finding that using a combination of pyridoxine deficiency and semicarbazide ad-ministration in vivo inhibits the whole body H F C of rats to within 10-20% of its normal value. They examined various tissues of the rat for histamine content every day for 90 daysfoir inhibition of histamine f o r m a -tion. In tissues like abdominal skin and tongue, which are r i ch in mast cells, inhibition of histamine formation to a small fraction of its normal rate for several weeks was not followed by a significant drop in histamine content; but in the gastric mucosa, which is a predominantly non-mast cell tissue, the histamine content fell to low levels within 24 hours of the start of the experiment (r7*8., 1£1). This was taken to indicate that in mast cell tissues, a histamine molecule, once formed, is f i rmly held and has a long intracellular lifetime; whereas in the gastric mucosa the reverse applies. In the rat lung which is believed to be poor in mast cel ls , the H F C was found to be relatively high. The histamine content was low and its lifetime exceptionally long. This situation indicates that the majority of newly-formed histamine is pooled so as to be able to leave the cell easily. A small portion appears to be retained in cells that possess a type of binding which provides for a long lifetime. H a r r i s et a l . (i!31') discovered that high concentrations of histamine occur in cerebral regions related to the hypothalamus and hypophysis, especially in the hypophyseal stalk. White (-187) showed that the cat brain forms histamine.by perfusing it with radioactive 144 histidine. These experiments marked the beginning of the investiga-tion into the occurrance and distribution of histamine in the whole brain and its regional tissues. White (13£ ) determined the in vitro H F C in minced brain tissue of the cat, pig, and dog and found that in the three species studied, H F C was highest in the hypothalamus. He also found that r ing N-methylation is the principal pathway for histamine catabolism in cat brain in vivo. It is l ikely that almost all histamine, at least in mammals such as guinea pig, rat, baby rat, rabbit, mouse, chick ? and frog (188 ), occurs in neural tissues and not in mast cells , for mast cells are not found in the central nervous tissues of those species that have been examined ( ]£? ' , 164). The rabbit, which is deficient in mast cells , has less histamine in the posterior lobe of its hypophysis than have other species. It would appear too, that the histamine of the anterior lobe of the hypophysis, though it is not in mast cells , is held differently f rom histamine in the b r a i n . This is because reserpine, chloropromazine and iproniazid did not change the levels of histamine in the hypophysis while they did alter levels in brain (1134}. Schayer came up with a new theory for the physiological function of histamine under some normal conditions and various abnormal conditions in laboratory animals. He was impressed by the fact that in a variety of circumstances an art if icial ly induced increase in histidine decarboxylase activity is accompanied by changes in tone and permea-bility of terminal blood vessels . The correlation between changes in 145 decarboxylase activity and microci rculat ion was the basis for the hypothesis that induced histamine, serves as a governor of the functional state of the terminal vessels . In order to understand more fully the relationships between these things, the following experimentally supported postulates are required; viz , histamine is continuously pro-duced within the smooth muscle and endothelial cells of small blood vessels ; histamine acts p r i m a r i l y on intrinsic or intracellular receptors , and the mechanism for histamine production can adapt to environmental requirements ( i . e . histidine decarboxylase is an inducible enzyme). According to these postulates, histamine is formed within smooth muscle cells , and diffuses out through the cell wall . A moderate excess of histamine then increases nutritive blood flow and permits cells to reach their full potentialities. Conversely, a histamine insuffi -ciency wil l lead to impaired cell nutrition and development of abnormali -ties in cell chemistry and function ( Kahlson (?£.) demonstrated extremely rapid formation of histamine during pregnancy in the rat . It was also shown that fetuses of the rat contained an extremely active histidine decarboxylase ($3). This led to the hypothesis that the process of histamine formation was in some way connected with rapid growth. This hypothesis is further supported by observations made on wound healing and liver regeneration (8Z). In an attempt to identify the tissues responsible for the increased formation of histamine during pregnancy in the rat, it 146 seemed to be logical to study the effect of removal of the fetuses on the urinary excretion of the amine. On removal of the fetuses, ur inary histamine fell sharply and rapidly reverted to the nonpregnant level or even lower, indicating that increased formation of histamine during pregnancy depends on the presence of the fetuses. Detailed studies of the sites of histamine formation in the fetus in vitro revealed that the fetal l iver is the most important location for histamine produc-tion: the H F C exceeds that of adult l iver by up to 1, 000 times or even m o r e . The histamine content of the fetal tissues was low, even below that of the mother, . in spite of the high tissue H F C . Here is a situation characterized by high H F C , but low histamine content and low capacity to bind histamine. These factors suggest that nascent histamine is involved. Other fetal tissues also displayed histidine decarboxylase activity at much higher levels than those found in the tissues of young or adult animals This was also confirmed by other investigators (25, 87')- A s pregnancy of the mouse proceeds, histamine excretion r i ses to values up to 100 times the nonpregnant value. After delivery, the urinary histamine falls , at f irst steeply, then more slowly, and the nonpregnant level is not attained until a few weeks after del i -very Striking differences exist in the regional distribution of histidine decarboxylase in fetal mice and rats: in the mouse fetus the H F C resides mainly in the skin, whereas in the rat fetus the l iver is the principle site of histamine formation. In the pregnant hamster, the site of elevated histamine formation is not the fetus but the placenta 147 The histidine decarboxylase activity determined isotopically in vitro in the extra-uterine tissues of the pregnant hamster (abdominal... skin, lung, l iver , spleen, gastric mucosa, small intestine, kidney and skeletal muscle) did not show significantly elevated histamine for-matiori -08$). Studies of histamine formation in the human embryo have been far less comprehensive than in the other species. Blood was collected f rom the umbil ical artery and vein while the embryo was in place, and then from a carotid artery after the embryo had been removed. P l a s m a histamine was found in significant amounts in the umbil ica l artery and in the embryonic carotid artery. Maternal plasma, like human plasma generally, was devoid of histamine in measurable concentration. There was more histamine in the umbil ica l artery plasma than in the plasma of the umbil ical vein ( 90). More direct evidence of histamine formation in human embryonic tissues was ob-tained by isotopic in vitro determinations (:*?!)). Most of the fourteen embryonic tissues examined displayed histidine decarboxylase activity, although at rather low levels ; highest in order of magnitude were spleen, hypothalamus, stomach, abdominal skin, and l i v e r . Healing skin and some tumors show enhanced formation of histamine. The rate of wound healing, as expressed in the tensile strength of the wound and the rate at which collagen formed in it, was measured in the normal state and after the H F C of the skin had been 148 ar t i f ic ia l ly lowered or elevated ( 93, 94 :, 95.). The histamine content of wound and granulation tissue is very low in comparison with that of normal skin. These tissues then, share with embryonic tissues the properties of H F C and high turnover rate of histamine. Histidine de-carboxylase activity in human skin has been demonstrated isotopically (19.6)-. This change is not caused by mast cells because the number of mast cells is not increased in the region of the wound. Lindel l et a l . ( ^ 1) also found the H F C to be low in normal human skin, but high in healing skin wounds. Injection of " long-acting" histamine, however, did not enhance the rate of healing, nor did injections of a histamine antagonist retard the normal process of healing (.93). Thus, the role played by nascent histamine cannot be served by extracellular histamine nor eliminated by antihistamine agents. After a high H F C had been recognized as being concomitant with certain kinds of rapid tissue growth, it appeared likely that a s imilar phenomenon might also be found in malignant tissues. Ri ley et a l . found that female rats bearing a subcutaneously implanted hepatoma had markedly elevated urinary histamine excretion levels which returned to normal immediately after removal of the tumor ( 9 8 ) . Kahlson et a l . have investigated various tumors; in the E h r l i c h asci tes tumor in mice they determined mitotic index, growth curve, H F C , histamine binding capacity and histamine content ( (08). A special inquiry into H F C , as it is related to growth, has been conducted using two different kinds of 149 tumors. A h l s t r o m et a l . (100) examined a transplantable, v i r u s -induced rat sarcoma and the enlarged liver of the tumor-bearing host. The H F C of the tumor was high; there was a tendency for higher H F C to be associated with more frequent mitosis and younger tumors tended to have a higher H F C . A rat Walker carcinosarcoma studied by Johnston U6I") grew rapidly, and the liver became enlarged. The H F C of the tumor and enlarged liver was high. That histamine is a powerful stimulant of gastric secretion was discovered first in experiments on dogs conducted during 1919-1920. A s imilar action in humans was demonstrated in 1922. It was subsequently shown that histamine stimulates gastric secretion in al l species that have been adequately tested. The work on this subject up to 1963 has been reviewed extensively by Ivy and Bachrach (102-') and by Code (\0ih)f" The gastric mucosa of al l species studied (man, dog, cat, rat, guinea pig, hamster, mouse, and frog) are r i c h in histidine decarboxylase (f7d). In the rat, mouse, and cat, mucosal H F C is much higher than in the other tissues, and is exceeded only by the high levels in certain tissues of pregnant animals and some malignant tissues. Resumed feeding of fasted rats, mice , and frogs evokes a mobilization of histamine and concurrent acceleration of the rate of histamine formation in the parietal region of the gastric mucosa. These changes in mucosal histamine are not merely a product of secretory activity since they do not occur when acid secretion is 150 excited by injection of histamine. Nor do they occur in regions of the mucosa that are devoid of non-parietal cells (18). Feeding, or the principal individual excitatory components operating subsequent to feeding, such as vagal excitation, production of gastrin, or distention of the stomach wall , induce an accelerated rate of histamine formation that persists for several hours (78). This sharp and long-lasting elevation of histidine decarboxylase activity is believed to be brought about by an initial lowering of the preformed histamine content of the mucosa, with the levels of the enzyme and the end product interrelated by feedback coupling. Using fluorescence microscopy to visualize gastric mucosal histamine in the rat, Thunfeerg; found fluorophores of histamine in cells located at the base of the gastric gland and in the few submucosal mast cel ls , 2.. . In ser ial sections, H F C was shown to be a property of that part of the gland which contains histamine. The distribution of parietal cells appeared to be unconnected with the distribution of histamine and the enzyme which synthesizes it. Histamine approaching the parietal cell f rom the outside causes the cell to secrete H C l , as seen from the stimulatory efficiency of injected histamine. In sections of the stomach, the number of acid-seer eting cells parallels the amount of histamine held in enterochromaffin-like cells in the gastric mucosa. Fur thermore , the output of histamine into the gastric juice parallels the volume of hydrochloric acid secretion whether this secretion is 151 induced by injection of gastrin, ingestion of food, or injection of cholinergic drugs. CAfcBACHOL M6THACHOHW .1 ( VAGUS Q-AVT-RIM Figure 13. Schematic representation.of .the approach of histamine to the parietal cell of the stomach (ref. 94>). The phenomenon of anaphylaxis was described by Port ier and Richet (1902) in a dog receiving sub-lethal doses of an extract of salt-water Actinies It appeared to them that instead of being . . " immunized", the dog developed a condition just the reverse , i . e . it developed a hypersensitivity to the toxic agent. The name "anaphylaxis" (Gk. ana-inverse, phylaxis-protection) was coined to convey this idea of "reverse^, immunity" . In 1910 Dale and Laidlow (.1.1:2..) f irst suggested that histamine might be involved in anaphylaxis. Release of histamine in anaphylaxis has recently been extensively reviewed by Rocha e Silva •> ( \0t) and several others ( lO?,. < 10S"? \CR > M O , U.H). Anaphylaxis in most animal species and human allergy, regardless of the precipitating 152 antigen, involve s imilar physiological reactions: increased capillary permeability (hence the development of edema and even hemorrhage), hemodynamic changes such as a marked fall in ar ter ial blood pressure , and contraction of smooth muscle ( 12/J, 128). These endogenous observations have provided considerable impetus to the search for endogenous principles of anaphylaxis. F o r some years it was thought that histamine was the only mediator of anaphylactic reactions, but more recent study indicates that histamine alone cannot account for al l the symptoms of allergic manifestations. In 1941, Dragstedt defined anaphylaxis as a phenomenon of "auto-intoxication" by physiologically active substances. Besides histamine,which still remains the most important chemical mediator, many other endogeneous principles such as heparin (11.2), serontin (5-hydroxytryptamine), polypeptides such as bradykinin, and an unidentified slowly reacting substance (SRS) have been implicated in anaphylaxis in animals (J06). Acetylcholine (113 ), catecholamines, and adenyl compounds may be involved to a lesser degree. The precipitating factor in allergy and anaphylaxis involves the interaction of two macromolecules : an antigen and an antibody. A n antigen may be a polysaccharide, protein, or a simple compound that is covalently bound to protein as defense against the specific antibody (I IS"). The mammalian organism manufactures a specific antibody f r o m the globulin fraction of the blood. Renewed contact of antigen and 153 antibody at a later date initiates a chain of events resulting in release of histamine and other compounds such as serotonin, SRS and bradykinin f rom mast cells (lo&j Il6 ), basophilic leucocytes, and platelets. Some agents that selectively release histamine and other substances from mast cells are : antigen-antibody, cationic proteins, amines (d-tubocurarine, stilbamidine, morphine, polymixin B) and compound 48/80. In the selective reaction, the constituents of the granules are rreileased without any apparent damage to the cell membranes '(I'M; Non-selective agents that release substances f rom mast cells are proteolytic enzymes, venoms and tissue injury. There are many indications that the release of histamine from the sensitized guinea pig lung is blocked by anoxia, low temperature, inhibitors which presumably act by antagonizing enzymatic mechanisms, and removal of C a ions. Renewed contact of antigen and antibody involves the fixation of bivalent immunoglobulin and activation of processes leading to the release of vasoactive mediators f rom the c e l l . In a study of the anaphylactic reaction in isolated guinea-pig tissues based on histamine release and Dale-Schultz type responses OU^). Mongar and Schild proposed a general scheme for a cellular anaphylactic histamine release mechanism which involves interaction of the antigen-antibody complex with a "heat-sensitive respiratory enzyme p r e c u r s o r " . Ca ions and a narrow p H range are required for this reaction, which is inhibited by SH reagents and other enzyme 154 inhibitors ( II4? , \2>0 ). This reaction produces the "active enzyme", which, in turn, is inactivated by a number of structurally unrelated compounds such as phenol, cyanide, cinnamate, and some antipyretics. It is this active enzyme that liberates bound histamine. Mota (|I7) proposed that the mast cells play a very important role in guinea-pig anaphylaxis, due to the high sensitivity of this species to histamine; but he believes that mast cells probably play an insignificant role in rat anaphylaxis due to the low sensitivity of this species to histamine and 5-hydroxytryptamine. Bronchospasm is the most important symptom of anaphylaxis in the guinea-pig. In the normal state, the guinea-pig tissues examined exhibited H F C in order of de-creasing magnitude as follows: small intestine, lung, spleen, l iver , aorta, uterus, skin; but according to Kahlson et a l . ( I T9), tissues with part icularly high H F C in the normal state do not have correspondingly high levels in anaphylaxis, and vice versa . Nor do the tissues in which anaphylaxis is predominant (for example, guinea-pig lung and rat intestine) exhibit strikingly high elevations of H F C . It is now known whether there is a parallel between sensitivity of tissues to the effects of intracellular histamine generated by elevated H F C and sensitivity to histamine acting from outside. Kahlson et a l . c laimed that in some tissues that are poor in mast cells , the observed elevation of H F C during anaphylaxis is on the same scale, or even greater, than it is in tissues that are r i ch in mast cel ls . Histamine formed during 155 anaphylaxis in the rat and the guinea-pig is , in their view, largely of non-mast cell or ig in . The discovery that histamine is not merely released, but is also newly formed in hypersensitivity and anaphylaxis reactions, caused investigators to look at histamine in a new way again. Histamine formed at a high rate and acting intracellularly may at least partially account for manifestations which formerly were not believed to be associated with histamine. In dogs, the more pronounced effects of anaphylatic shock are hypotension, portal venous constriction and hemorrhage of the G-I tract. It has been demonstrated that histamine is also released in human allergy and anaphylaxis (12^), which conditions may be manifested by a combination of ur t icar ia , hypotension, and broncho-constriction (.l-tOVTfl'j). L i l j a et a l . found that human lung tissue forms histamine, i . e. it is endowed with H F C . The comparatively low rate of formation was of the same order of magnitude as that seen in the skin of normal rats ( 130). The discoveries that increased histamine formation takes place in al l tissues investigated in the anaphylactic animal, and that the elevated H F C persists for a long time after histamine release has ceased, seem to be helpful in explaining the failure of histamine antagonists to afford protection in the later stages of anaphylaxis. Histamine antagonists may not interfere with actions caused by histamine formed and acting within tissue ce l ls . It is presumed that in anaphylaxis, 156 agents other than histamine (serotonin etc. ) are responsible for symptoms which are not alleviated by histamine antagonists (T0(>). The histamine antagonists do not interfere with the important actions of endogenous histamine in gastric secretion and in the metabolic processes of tissue growth and protein synthesis. This is presumably because histamine is not engaging receptors of the familiar sort in these functions. The limitation is that the feed-back relation between histidine decarboxylase and histamine, by its very nature, is likely to exclude the feasibility of inhibiting histamine formation entirely. Endeavours of the last ten years have not succeeded in demonstrating histamine release in any purely physiological event. One case of physiological histamine release has been shown to occur : the histamine content of the gastric mucosa of a rat deprived of food is lowered when the animal is fed. This phenomenon was interpreted in terms of alterations in a chain of events in the mucosal histamine metabolism designed to stimulate and sustain the secretion of hydro-chloric acid. A solid body of observations provides the basis of theories which causally associate gastric mucosal histamine release and forma-tion with excitation of the parietal cells , and nascent histamine with the anabolic processes of growth and protein synthesis in various tissues. The foregoing is a summary of developments in some areas of histamine research . The desire to treat allergy more effectively is 157 the p r i m a r y reason for our interest in histamine. Table VIII is included with this in mind. It details the various stages of al lergic reaction and the treatments that can be given along the way. 158 Table VIII A C T I O N O F D R U G S , O N T H E A L L E R G I C R E S P O N S E (Ref. 192) Sequence of A l l e r g i c Reaction Antigen Antibody forming cells Ab + Ag Release of humoral agents Immediate response of target tissue: Vascular dilatation Broncho constriction Resulting disturbance in organism: Change in mucus Cough Edema and itching Lacr imation and rhinorrhea Insomnia.nervousness, exhaustion Hypoxia Hypercapnea Acid-base imbalance Shock Infection A n t i - a l l e r g i c Action: : Stage and Type Prevention of antibody formation Prevent formation of humoral agent (histamine) Stabilize lysosomes Depletion of humoral agents Interference with action of humoral agents Reverse undesired effects Medication A n t i - a l l e r g i c drugs Immunosuppr es sants Histidine decarboxy-lase inhibitor Steroids Histamine (et a l . ?) liberators A n tihi s t am in e; s et a l . Adrenergics Theophylline Supportive treatment Expectorants and mucolytics Antitussives Analgesics Tranqui l izers and sedatives Oxygen Acid-base buffers Vasopressors Antibiotics 159 P A R T VI H I S T A M I N E " R E C E P T O R S " Little is known about the " receptors" for histamine in the animal body, but two facts can be deduced from studies of structure-activity relationships and the effects of specific blocking agents (7). F i r s t , wherever histamine receptors are located in the body (in smooth muscle , capillary endothelium, nerves, gastric glands, etc. ), they are distinct f rom each of the other " receptors" activated by other autacoids like the catecholamines, 5-hydroxytryptamine, acetylcholine, bradykinin and angistensin. The other thing known for certain about histamine receptors is that there is more than one type. The specific antagonism of some ractions of histamine by low concentrations of antihistaminic drugs characterizes one type of histamine receptor, to which A s h and Schild (174) give the symbol H ^ . . Receptors of this type occur in guinea-pig ileum and bronchus. Other actions of histamine like: stimulation of gastric acid secretion and stimulation of isolated rat uterus are not antagonized by common antihistaminic agents (775). These actions are likely to be mediated by receptors other than those designated Hj, . The receptors involved in these actions are designated as H ^ -K i e r (176) proposed that two conformations of histamine are preferred ; one with the tertiary nitrogen of the side chain and the 160 pyridine nitrogen of the molecule (a) 4. 55 A apart, the other (b) with these nitrogens separated by 3.60 A ° . He suggested that the \ / C — M H H • M 1 T 1 H / H H 1  (a) 6CC = |80», £v-.-^ - C =120" (b) Sec = 3 0 0 ° , © i - ; ^ - C =120" ' F igure 14. ConformationsJof histamine conformers attach to different types of receptors, H^ and H ^ . K i e r , therefore, postulated that two distinct biological responses depend on the presence of one or the other complementary receptor. The H^ receptor would be complementary to the internitrogen relationship (a) in F i g . 14 , whereas H-, receptor would be complementary to the internitrogen relationship (b) in F i g . 14 . K i e r ' s calculations were based on the extended Huckel method for molecular orbital calculation. Green et a l . (177) severely cr i t i c ized K i e r ' s work,saying that the Huckel method gives "hopelessly inaccurate resul ts" and is "worthless as a procedure for predicting the structure or chemical behaviour of molecules . " In K i e r ' s work, too, the dihedral angle was rotated every 6 0 ° , a procedure that could miss important conformers . Green et a l . used the INDO (Intermediate Neglect of Differential Overlap) molecular orbital method. The total energy of the molecule was m i n i -m i z e d as a function of two dihedral angles, ^ and 0 ^ ^ for 161 histamine. The angles were rotated at 3 0 ° intervals and at 1 5 ° when 4. <* N r r CH* - C H 2 ~ W H * this was necessary. The result is that 98% of histamine free base should exist in the configuration with © , „ = 1 5 0 ° and © . „ = 3 3 0 ° . In this configuration, a suitable distance occurs between one of the side-chain amino hydrogens and N-3 of the imidazole ring to permit hydrogen bonding. Analysis of the histamine cation showed the con-former with p = 1 8 0 ° (in the plane of the imidazole ring) and ©^ p = 3 0 ° or 3 3 0 ° had least energy. . The quaternary nitrogen in this structure was close enough to N - 3 of the imidazole ring to permit hydrogen bonding. Pullman et a l . (178) confirmed this result by finding that the folded structure of the cationic form of histamine at physiological pH most stable; it included an intramolecular hydrogen bond. A l l this information about the conformations of histamine has done very little for our understanding of the receptor for histamine. One way to conceptualize the receptor is to speak in general terms and say that the receptor for a cationic molecule should incorporate an anionic part so that ionic bonding is possible. Since the action radius of ionic groups are relatively large (the electrical field around the ions decreases with the square of the distance), it is postulated that the 162 amino group serves as a guiding group leading the drug to its receptor site, and then the relatively weak forces whose action radi i decrease with the seventh power of the distance, hydrogen bonding and Van der Waals forces, come into action and stabilize the complex and make possible a close fit at the receptor surface (179). Another model for the histamine receptor was suggested by Rocha e Silva (106). According to his scheme, histamine attaches to its receptor by means of two sites: the imino group of the imidazole ring and the free amino group of the side chain. The imino group forms a transitory bond with the polarized carbonyl group of a peptide linkage of the receptor; the second one forms a hydrogen bond with a protein residue of histidine or arginine. Other forces, mainly hydrophobic ones, may assist in the interaction. F igure 15 . . Schematic representation of histamine receptor (ref. 106). 163 P A R T VII A N T I H I S T A M I N E S Antihistamines are drugs with the ability to antagonize in varying degrees most, but not a l l , of the pharmacological actions of histamine (179). They seem to share with many other pharmacolo-gical antagonists the property of competitively occupying the receptor sites on the effector cells to the exclusion of the agonist. This implies that the effects of the presence of a certain amount of histamine wil l be countered by a sufficiently high dose of an antihistamine, and vice v e r s a . Antihistamines apparently bind with the histamine receptor without initiating a response. In this manner, they avoid the full impact of increased histamin forming capacity (HFC) and the resulting presence of elevated concentrations of histamine in the blood and tissues. A n t i -histaminics diminish, in varying degrees, the bronchiolar and intestinal spasm, increased capillary permeability, salivation, cutaneous wheal, and release of epinephrine f rom the adrenals caused by histamine. How-ever, antihistaminics have no effect on gastric secretion induced by histamine. A new class of compounds which competitively inhibit actions of histamine on gastric secretion has recently been reported (174). . The symptoms of elevated histamine content of tissues may also be alleviated by the administration of physiological antagonists, e . g . , ephedrine or epinephrine, which have an effect diamtrical ly opposed to that of ' . histamine. S T R U C T U R E - A C T I V I T Y R E L A T I O N S H I P S A M O N G T H E A N T I H I S T A M I N E S M a x i m u m antihistaminic activity is found in the following structure, where R^ and R-, are aromatic or heteroaromatic r ings, one of which may be separated f rom X by a methylene group. One Figure 16. General structural formula of antihistamines of the aromatic rings may be replaced by an al icylic system, (e .g. cyclohexyl) without loss of activity (181, 6). X may be N , C H , or an oxy ether to carbon. ( ^ I C H - O - ) . The use of a thio ether reduces activity (5). The nature of X is commonly used to classify antihis-taminics . Except in cases where Y is present, R^ must be ethylene to attain good antihistaminic activity. Lengthening or branching the chain markedly decreases activity. The antihistaminics are then named as propylamine, ethylenediamine, or ethanolamine derivatives. Two subclasses of the ethylenediamines are the piperazine and pheno-thiazine derivatives. Examples of antihistaminics belonging to each class are given in Figure 16. R^ and R 2 may be linked in the ortho-positions by Y, which is a methylene, heteroatom, or methylene-heteroatom function. R^ and Rg are usually methyl groups, although - N ( R 4 ) 7 may be incorporated into a small ring system. The use of 165 ethyl groups for R and R5 decreases the antihistaminic activity. A distance of 5 - 6A from the amino nitrogen to the center of one of the aromatic rings gives the strongest competitive activity. Substitution of one of the aromatic rings with a p-methyl or p-chloro group generally increases activity. Ortho- substitution on one or both rings decreases activity. Nauta (2) has explained this relationship in alkylsubstituted diphenhydramine derivatives by proposing that the TC electrons of one of the aromatic rings interact with the oxygen lone pair electrons, resulting in an altered electron density on the oxygen atom. P a r a -alkyl substitution enhances the overlap interaction, while ortho- and meta- substitution decrease the interaction. Table IX A N T I H I S T A M I N E S 166 E T H A N O L A M IN E D E R I V A T I V E S Chemical Name (Generic; Brand) Chemical F o r m u l a Thymoxyethyldiethylamine ( ; 929F) CH, O-CH 2-CH 2-N-CH 2'T'CH 3 C H - C H 3 C H 2 — GH3 2-(Benzohydryloxy)-N, N -dimethylethylamine (Diphenhydramine; Benadryl) C H - O - C H - C H 2 - N - C H 3 C H 3 2- (p-bromo- ck -phenylbenzy-loxy)N, N-dimethylethylamine (Bromodiphenhydramine; Ambrodryl ) Br- C H - O - C H - - C H - - N - C H , 1 2 2 3 C H , 2-( ck -(2-Dimethylaminoethoxy)-- ^ -methylbenzyl)-pyridine (Doxylamine; Decapryn) ,—N C H C - 0 - C H 2 - C H 2 - N - C H 3 C H (2-Benzylphenyl)- /S -dimethyl-am ino - e thyle the r (Phenyltoloxamine; Bristamin) / / \ O - C H - C H - N - C H 2 2 J 167 T A B L E IX E T H A N O L A M I N E D E R I V A T I V E S (Continued) Chemical Name Chemical F o r m u l a (Generic; Brand) 2-(p-Chloro-ek - (2-dimethyl-aminoethoxy)benzyl)-pyridine (Carbinoxamine; Clistin) 0 - C H 2 - C H 2 - N - C H 3 C H 3 Piperidinomethyl-2-benzo-dioxane ( ; F933) C H 2 - N; ( ) ((llime^Kyo"-?^ C H I 9 7 \ \ _ C H - 0 - C H 9 - C H 7 - N H C H 0 M' 2 2 I C H , O-T A B L E E T H Y L E N E D I A M I N E D E R I V A T I V E S ( ) ( ; 2325RP) C H 3 - C H 2 - N - C H 2 - C H 2 - N - C H 3 N - b e n z y l - N - p h e n y l - N ' - N 1 -dimethylethylenediamine ( ; Antergan) 168 T A B L E IX E T H Y L E N E D I A M I N E D E R I V A T I V E S (Continued) Chemical Name i (Generic *;TBrand) Chemical F o r m u l a N - p h e n y l - N - ( 2 - t h e n y l ) - N \ N 1 -dimethylethylenediamine (Methaphenilene; Diatrin) C H 2 - N - C H 2 - C H 2 - N - C H 3 C H . 2 - (2 - dime thy 1 amino e thy 1 - (p-methoxybenzyl) amino) pyridine (Pyri lamine; Neo-Antergan) CH3-0-(/ \- C H 2 - N - C H 2 - C H 2 - N - C H 3 C H N - b e n z y l - N ' , N ' - d i m e t h y l - N - 2 -pyridylethylenediamine (Tripelennamine; Pyribenzamine) C H 2 - N - C H 2 - C H 2 - N - C H 3 C H , 2 - £ ( j3 - dime thylamino ethyl)-2-thenylamino^-pyridine (Methapyrilene; Thenylene, Histadyl) C H 2 - N - C H 2 - C H 2 - N - C H 3 C H , N ' , N , - d i m e t h y l - N - ( 2 - p y r i d y l ) - N -(5-chloro-2-thenyl)-ethylenediamine (Chlorothen; Tagathen) C H 2 - N - C H 2 - C H 2 - N - C H 3 N C H , T A B L E IX E T H Y L E N E D I A M I N E D E R I V A T I V E S (Continued) Chemical Name (Generic; Brand) Chemical F o r m u l a 2-(2-dimethylaminoethyl)p-methoxy-benzyl) amino) pyrimidine (Thonzylamine; Neohetramine) C H 3 - 0 ^ 7 V H C H 2 - N - C H 2 - C H 2 - N - C H 3 C H . N N 1 , N ' - d i e t h y l - N - e t h y l - N - p h e n y l -ethylenediamine ( ; 1571F) C H 3 - C H 2 - N - C H 2 - C H 2 - N - C 2 H 5 C 2 H 5 ( (Thenyldiamine; Thenfadil) // \N C H , - N - C H . - C H . - N - C H . C H , T A B L E P R O P Y L A M I N E D E R I V A T I V E S N , N-dimethyl-3-phenyl-3- (2-pyr idyl)-propylamine (Pheniramine; Trimeton) C H - C H , - C H , . - N - C H _ 2 2 | i 3 C H . 2 - (p-chloro- tf( (2-dimethyl-aminoethyl) benzyl)-pyridine / —N (Chlorpheniramine ; C h l o r - T r i m e t o n ) \ / (Dexchlorpheniramine; Polaramine) C H - C H 2 - C H 2 - N - C H 3 CH-, 170 T A B L E IX P R O P Y L A M I N E D E R I V A T I V E S (Continued) Chemical Name  (Generic; Brand) 2- (p-bromo- - (2 -dimethyl-aminoethyl)benzyl)-pyridine (Brompheniramine; Dimetane) (Dexbrompheniramine; Disomer) r— N <^ y - C H - C H 2 - C H 2 - N - C H 3 Chemical F o r m u l a 1 - i^:-(p-chlorophenyI)-3-phenyl-2-butenyl^-pyrrolidine (Pyrrobutamine; Pyronil) C H , V / / • C = C H - C H 2 N' C l - . C H , T r a n s - 2 - ( 3 - ( l -pyrrol idinyl) -1 -(p-to lyl)propenyl)-pyridine (Triprol idine ; Actidil) , — N <^  ^>~C=CH-CH, — IM C H -T A B L E P H E N O T H I A Z I N E D E R I V A T I V E S 10- (2-dimethylamino- 1 -propyl)-phenothiazine (Promethazine; Phenergan) C H 3 C H _ - C H - N - C H o 2 , 3 C H . 171 T A B L E IX P H E N O T H I A Z I N E D E R I V A T I V E S (Continued) Chemical Name Chemical F o r m u l a Gener ic ; Brand) 10-(2-( l -pyrrolidyl)ethyl) -phenothiazine (Pyrathiazine; Pyrrolazote) / / \ \ N— C H 2 - C H 2 - N' 10 - (3 - dime thy lamino - 2 - me thy 1 propyl )phenothiazine (Tr imeprazine ; Temari l ) C H M - C H 2 - C H - C H 2 - N - C H 3 C H 10-(l -methyl-3-pyrrolidyImethyl)-phenothiazine (Methodilazine; Tacaryl) / / \ \ N - C H -N - C H , 10-(2-dimethylaminopropyl)-9-thia-1, 1O-diaza-anthracene (Isothipendyl; Theruhistin) C H , -CHo-CH-N-CH-: I C H , T A B L E IX (Continued) X Y C L I Z I N E S A N D O T H E R D E R I V A T I V E S Chemical Name (Generic; Brand) Chemical F o r m u l a 2- (N-benzylanilinomethyl)- 2-imidazoline (Antazoline; Antistine) C H 2 - N - C H 2 - C 1 -diphenylmethyl -4 -me thy lpipe r a zine (Cycl izine ; Marzine) 1 - (p-chlorobenzhydryl ) -4 -methylpiperazine (Chlorcycl izine ; Perazil ) - C H N - C H , 1 - (p- chlorophenylbenzyl ) -4-(m-methylbenzyl)-piperazine (Meclizine; Bonamine) 2 -methyl -9 -phenyl -2 , 3, 4, 9-tetrahydro-1 -pyridinedene (Phenindamine; Thephorin) C l C H , \ / 2 \ -N - C H , ! 173 T A B L E IX C Y C L I Z I N E S A N D O T H E R D E R I V A T I V E S (Continued) Chemical Name Chemical F o r m u l a (Generic; Brand) 2 - (1 - (2 - (2 - dimethylamino ethyl)-3 -indenyl)e thy 1)-pyridine (Dimethpyrindene; Forhistal) > C H 2 - C H 2 - N - C H 3 / / \ ) - C H - C H 3 C H -174 C O N C L U S I O N Nauta et a l . have reported the significance of overlap interactions (homoconjugation) between heteroatoms like oxygen and sulphur and the TC electrons of the phenyl group in diphenhydramine and thiodiphenhydramine. These interactions are modified by mesomeric , inductive, and steric effects. . Changing the amount of interaction changes the antihistaminic activity ( £ , S ). The following compounds were prepared to allow more information to be obtained regarding the effects of homo- and p T C conjugation and lack of conjugation on the antihistaminic activity of antergan analogues: N , N - d i m e t h y l - N ' - c y c l o h e x y l m e t h y l - N ' - o -methyl -phenylethylenediamine; N , N - d i m e t h y l - N ' - c y c l o h e x y l m e t h y l - N ' - m -methylphenylethylenediamine; N , N - d i m e t h y l - N ' - c y c l o h e x y l m e t h y l - N * -p-methylphenylethylenediamine; N , N - d i m e t h y l - N 1 - cyclohexylmethyl-N'-p-bromophenylethylenediamine; N , N - d i m e t h y l - N ' - 3 - c y c l o h e x e n y l -methyl-N'-cyclohexylethylenediamine; N , N - d i m e t h y l - N ' - 3 - c y c l o -hexenylmethyl-N'-phenylethylenediamine; N , N - d i m e t h y l - N ' - d i p h e n y l -methylethylenediamine; N , N - d i m e t h y l - N ' , N'-dibenzylethylenediamine; N , N-dimethyl -N' -benzyl -N' -cyclohexylethylenediamine ; N , N - d i m e t h y l -N 1 -cyclohexylmethyl-N'-cyclohexylethylenediamine. These analogues were prepared by reacting the appropriate p r i m a r y amines with acid chlorides like <<-chloroacetylchloride and benzoyl chloride, to form 175 °k - chloro ace tamide derivatives and benzenecarboxamide. The oC -chloroacetamide derivatives were then substituted nucleophilically with dimethylamine to form c<-N, N-dimethylaminoacetamide d e r i -vatives. The aminoacetamides and N ' - ( / - N , N-dimethylaminoethyl)-benzenecarboxamide were then reduced with lithium aluminum hydride to form the ethylenediamine derivatives. The ethylene diamines were reacted with the following carboxylic acid chlorides: cyclohexane-carboxyl , phenylcarboxyl, 3-cyclohexenecarboxyl, and 1-cyclohexene-carboxylchloride in the presence of triethylamine to obtain the N l-(fi -N , N-dimethylaminoethyl)-carboxamide derivatives. The carboxamides were also reduced with lithium aluminum hydride. This sequence of reactions generally gave a good yield for al l the analogues. A n excep-tion was the p -bromophenyl derivative. A worse problem was the reduction of the olefinic bond in the oC , fi -unsaturated aminocar-boxamide, N ' - c y c l o h e x y l - N ' - ( fi - N , N-dime thy lamino ethyl)- 1 - cyclo-hexenecarboxamide: when this compound was reduced with lithium aluminum hydride, both the carboxyl group and the olefinic bond of the 1-cyclohexenyl moiety were reduced. N ' - 3 , 5-dimethylphenyl-N'- ( fi -N , N-dimethylaminoethyl)cyclohexanecarboxamide was also prepared, but subsequent purification and reductions were not completed in this work because of a time shortage. The preparation of the following cyclohexadiene analogues of antergan was attempted: N , N-dimethyl -2 , 5-cyclohexadienylmethyl-176 N 1 -cyclohexylethylenediamine and N , N - d i m e t h y l - N 1 - 1 , 4-cyclohexadienyl-. m e t h y l - N 1 - cyclohexylethylenediamine. 2,5- cy clohexadiene-1 - carboxylic acid was prepared by B i r c h reduction of benzoic ac id . The preparation of 1, 4-cyclohexadiene-1-carboxylic acid began f rom preparation of 3-chloro-4-cyclohexene-1 , 2-dicarboxylic acid anhydride using the D i e l s - A l d e r synthesis of 1-chlorobutadiene and maleic anhydride. Due to spontaneous aromatization of the cyclohexadiene acid when it was prepared, it would have been extremely difficult to isolate these acids in the pure f o r m . But 2, 5 -cyclohexadiene- l -carboxyl chloride was prepared in order to get N-cyclohexyl-2 , 5-cyclohexadienecarbox-amide. The reduction of this compound with lithium aluminum hydride resulted in reduction of the carboxyl group and oxidation (aromatization) of the cyclohexadiene system. Chloroacetyl chloride was also employed in the reaction of N-2 , 6-dimethylphenylcyclohexylmethylamine prepared in this work and commercial ly available diphenylamine. The product could not be isolated due to its instability; a tarry mixture was formed. Alkylation using cyclohexyl-p-toluenesulphonate^ 1-chloro-3-cyclohexene and chlorocyclohexane hWijth''"" N , N - d i m e t h y l - N ' - c y c l o -hexylmethylethylenediamine^and N'-benzylethylenediamine was an impract ical method of obtaining antergan analogues. A l s o , the tertiary amine site of ethylenediamine appeared to be more active than the secondary amine site of the molecule. Identification of most intermediates and final products was 177 done by i n f r a - r e d , nuclear magnetic resonance, elemental analysis. ' r . Elemental analysis of liquid amines and aminoacetamide derivatives were performed on hydrochloride, picrate, per chlorate, r .1 methyl iodide derivatives. 178 B I B L I O G R A P H Y R o m m , P . , Guryanova, E . N . , and Koche sko v, K . A . , Tetrahedron 25, 2455-2468, Pergamon P r e s s , (19691 Nauta, W . T h . , Rekker, R . F . , and Harmes , A . 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