ALKYLA-TION STUDQES WITH AMf-NOTRIAZOLES AND AMINOTETRAZGLES Thai: 50: {he Dogma of Ph. D. MEWiGAN STATE UNIVERSSTY Kenmfi Raiph Wilson 1957 f l THESIS ”r ' * a": "‘"W EAST LANSING, WHVI'ildAN LIBRARY MiChian S {arc University afieu , H O J V ‘ past wwmc, MICHIGAN "‘1. 3“?! L W110}! STUDIES WITH AMINOTRIAZOLES AND MNOTETRAZOLES By Kenneth Ralph'Wileon .1 THESIS submitted to the College oandvanoed Graduate Studiel of Michigan Stet. Univereity of Agriculture and Applied Science 1n.part1al fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Department of Chemistry 1957 EKNOWDGWT The author niche! to expreee hie uppeeeietion to Dr. Robert H. Herbet for hie helpful guidance and counsel througheut the coerce of this work. Appreciation 1e clue tendered to the Parke-Devil Company for financial eeeietance through the Perke-Devie Fellcuehip during the academic year! 195h~l955, 1955- 1956 and the fell tern of 1956-1957. 11 WIDE STUDIES WITH AMINOTRIAZOLES AND flflNOTETRAZWES By Kenneth Ralph Wileon AN ABSTRACT Submitted to the College of Advanced Graduate studiee of Michigan State Univereity of Agriculture and Applied Science in partial fulfillment of the requiremente for the degree of DOCTOR CF PHILOSOPHI Department of Chemistry Year 1957 Approved @WW/WW ABSTRACT Since the reaction of Wl-Soaflnotetraaolee with alkyl balm” and alkyl bmeeueeulronatee had been ehoen to lead to l,h~dialkyl-S- ininotetraaolinee (1,2) a My of the allqlation of h—avdno-l,2,h- trimlee me undertaken. When 3.5-dLnetrsyl-hml,2,h-triaeole we treated with bmayl— chloride a prochxct resulted which appeared to be a quaternary chloride. Thia maternal-y chloride when treated eith potaaeim max-oxide me an nine bale Ihich would tom a Weehloride ieomeric with the (“ternary chloride. Treatment 01’ the hue obtained from the quaternary chloride leada to a eecond quaternary chloride. The reactions described above an: be enmrieed by the follceing echemea 5...: x c 8 013 J! [I canes-cage; KGH(aq.) % 45-3“ on, \ Icaca, * c,,n,=cm.mc"n“w‘ ¥ ' 1 CH 1 m. Re .0 C‘HJ. . ciefieicme We When 3,5-diphetwl-hvm-rl,2,h-trieeole eae treated with beneyl chloride a quatermry chloride lee obtained that appeared to be um in nature to the chloride 013mm 2mm 3,5-dimethyloh-emim-l,2,h- m1. an beneyl chloride. Treatment at the diphenyl quaternary chlorideuithbaee gaveanoil tronehichapureprodncteeuldnctbe iaolated. iv Hethylation of 3.5vd1mt2vloh-Mm-l,2,h~triaeole and 3 ,S-di- phezwl-h-aMnooIJ ,h-triazole with methyl benzeneeulronate me attmtedg however, the products which were obtained were not curator.- iced. During the couree of thie investigation a mitivity to one or more of the compounda being handled forced a ceeeetion of further work Iith hem-1,2,h-triaeelee. Because of thie amitivity, the direction or the problem was altu‘ed to imlnde the arelkylation or l-cycloamyla-S-axzdnotetrmles V with the intention or preparing potmtiall: nomiclogiceny active, l-cyoloalkylnh-arancyl-S-Lfinotetrmline Willem”. A eeriee or l-cycloelkyl-h-aralkylaS-iadnotetraeoline hydrochloridee he prepared by trummt or locyclohuylv and l—cyclohexylnetrwl-S- aminotetraeole with hm]. chloride, pochlorobeneyl chloride, o-ehlnroa» bum chloride, 2,h—&ichlombml chloride, 3 ,h-diohlorobenayl chloride, p-dnitrcbenzyl chloride, u-nitrobeneyl chloride, Egg-phenylo etlvl bromide and maphem’lprcpyl bmmide. The” comma we characteriaed by formation of pherwlthicereu by reaction of the bee bale with phony]. ieothiooymte and, in some instances, by the ieolation o: the tree minotetraeoline as a metalline eolid. The atruohare o: the cocpounde m eetabliehed by the analog of the method or preparation eith that deeoribed for the preparation of commence of. known etrecture and by comariaon of their infra-red epectra uith epectra of eimiler comma of km etrnctnre (2). References Cited (1) R. A. Han-y, W. a. Finnegan and E. Lisber, J. Am. Chen. 800., 76. 2891: (1351;). (2) D. I. Pmim, 'Alkyhted SW1“ Their Preparation and Pmpertioe,‘ Ph. D. mania, Monica: State College, 1955. I%HUE(H’COMHQHB Page WIOOOOOOOOCOCCC0.00000QOOOOOIOO'OCCOCOOOO0.0.00.00.00.00... FIR? I mowmoxOOOOOOOOIOOOOOOOOOOOOOOCOOOOOOOOOOOIIOOIOOOIOIOCOOOOO DMSSIONOOOO...OOOOOOOOOOOODOOO‘..OOOOOOCOOOOOOOIOO0.0.0.000... mmnlOOOOCI00......COOCOOOOOOOOCOOOOOOCOOODIOOUOOOOOOOO0.0 hW‘fiOB 01‘ 3 'S-DiNbUtitutGd h-Amino-1,2,b-Triaeolelo e e e e e B’s-Diphew1.hw0°1,2,13‘tr133010eeeeeeeeeeeeeeeeeeeeeeee 3,S-Dimethyl-h-aminO-l,2,h-triamle.o"noun”...no...” Alkyietion of 3,5;Dienbetituted h-Amino-1,2 horrieeolee....... Methyletion of 3,S-diphenylph-amino—1,2, -triazole......... Hethyletion of 3,S-dimethylph-emino-1,2,hetriezole......... Beneyletion of 3,S-dimethylph-emdno-l,2,h-trieeole......... Emotion Of Homelkyletod Product. With MCeeeeeeeeeeeeeeeeee treetnent of methylated 3,5-dinethylrh-elino-1,2,h~trieeole Elf-h meeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee Treatment of beneyleted 3,Sidiphenylphnenino-l,2,hptrieeole llth “.0...eeeeeeeeeeeeeeoeeeeeeeeeeeeeeeeeeeeeeeeeeeee Treetnent of bennyleted 3,5;dinethylph-emino-1,2,h~trieeole '1“! buCeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee Benlyletion 0: Compound Beeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee Reaction 0: CW D With Moeeeeeeeeeeeeeeeeeeeeeeeeeeeee WWI-”1| Of Benxyleted MWeeeeeeeeeeeeeeeeeeeeeeeee Attempted debenxyletion of 60W Beeeeeeeeeeeeeeeeeeeeee Debennyletion Of compound leeeeoeeeeeeeeeeeeeeeeeeeeeeeeeee Continuation Of h‘W’1,2,h.mlu with “demueeeeeeee Condeneetion.of aromatic eldehydee Iith 3,S¢dimethy19h- wo'lgzgh’tmIOJ-Coeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee Condensation of‘beneeldahyde Iith h-elino-1,2,h~trieeole... Reaction of 3,SQdimethylphsemino-1,2,hdtriesole with formic acid-formaldehyde."on-nunuuuun.o.’........uu. HydrOgenetion or Beneeldehyde-hqemino-1,2,hprrieeole Mixturee. Hydrogenetion of banzaldehyde—B,S-dimethyl-h-emino-l,2,h~ tm‘OhCCCOOOOOCC.0.......................O...‘........ Attempted hydrogenation of the benealdehyde h-emino-1,2,h- W501. condensation pmdmteeeeeeeeeeeeeeeeeeeeeeeeeee v11 x1 TABLE OF COHTENTS - Continued Page Reeetion or Hitroue Acid eith 3,5-Dinethyl~h—emino-1,2,h- m1. Ind I“ Emulation Proaflueeeeeeeeeeeeeeeeeeeeeeee Reaction of 3 ,S-dinetlvl-h-Mno-lfl ,h-trieeole eith nitrate ‘01deeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee Reaction 0f Compound 1 With nitroue Icideeeeeeeeeeeeeeeeeeeee Reection of Compound C “ith nitrOU. Icidqeeeeeeeeeeeeeeeeeeee Infra-red Abnorption Spectr‘eeeeeeeeeeeeeeeoeeeeeeeeeeoeeeeeeeee PART II INTRODUCTIONOCOOOOOOOO..00....OOOOODOOOOOOCOOCO.0.0.0.0.0....COO... DISCUSSIONOOOCCOOOOQCOOI.0.O....O'OOOCQOIIO'COOCCIOOOOOOOOOOOOIOOO. EIPERIMENTALOOOOOUOO.QOCCOCOOOOOOOOOIOC.00....OCOOOOOOIOOOOOOOOQCI0 Preparation 0: GyCIOthylflchylflminfleeeeeeeeeeeeeeeeeeeeeeeee lellkylvselninotetreeolee.....g................................. 1»CyclohexylmethylpS-eminotetreeole....o....o................ 1,h-Dieubetituted S-Iminotetreeoline Hydrochloric!” . . . . . . . . . . . . . 1-0yolohmql—h-p—ehlombemyl—5—Mnotetruoline hydro- chloride-eeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee 1-Cycloheoqlmethylnhob efl-phewlethyl-S-Mnotetreeoline hydrOChlorideeeeeeeeeeeeeeeeeeeeoeeeeoeeeeeeeeeeeeeeeeeeoe DGT1'.tiVOI With Phenyl Ieothiooylneto.......................... thlthiouree derived from l-cyclohexyI-h-p-chlorobeneyl-S- 1N1nOtCtr‘3011neeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee lrcyolohexylmathyl~h-erelkyln5-in1nototrezoline!.eo......a.....o Infr‘Pred ‘b'orption.SPQOtr.eeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee SUHM‘RIOCIOC.O00.00.0000...O...C...OOOOOOOOOOOOOOOOOIOCQOOO00...... LITER‘IURE CITED.OOOOOOOOOIOOOOOOOO...0.0..OOOOICOICIOOOIOOOOOUOCOU "PEHDIIODOOOOOOOCOOIOC000......OO‘OOOOOOOOOO...IOOOOUOOOOOOOOOOOOO viii 31 3.1 31 32 .32 33 35 145 h? ha h? 5h 5h 58 59 62 rm VI VII LIST OF TABLES Page lmhhmbhwdkyloS-Mnotetreeoline Hydrochloridee . . lmhhwhetlvl-hdrflkylmetreWMne Hydro- ' ChloridOCeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee mm: of 1-Cyn1oheaql-h-ere1ky1-Soininotetruoline HydrOChloridQ‘eeeeeeeoeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee Anaheie of I-MIohmmum-hqrflkyl-S-ininotetn- [Olinfl Hydrochlorid93eeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee Phenylthioureee Derived from 1-0yulehay1-u-nrakyi-s. iminototraeoline Hydrochloridea....o.o.......a............ thylthionreee Dan-ed from 1-Cyeloheacy1methy1-bu erelky1e5¢1minotetreaoline HyerChlorida‘eeeeeeeeeeeeeeeee IWhmml'hml‘S‘momuam e e e e e e ‘. e e e 50 52 53 SS LIST OF FIGURES FIGURE Page 1. Intro-red Spectra of 3,5—Dimethy1—h-muno-1,2,h-trieeole, 00W A, Compound B, Compound 6 end Compound Dunn”... 8 2 . Infra-red Spectrum of 1-Cyclohemy1ah-b snebe-ininotetreo eoline Hydrochloride....o...............o...o...s.......... 62 3 . lune-red Spectrum of. 1-Cyclohuq1-h-p-chlorobeney1-S- ' ‘ ininotetreeoline Hydrochlorid‘eeeeeeeeeeeeeeeeeeeoeeoeeeeee 63 h. Infra-red Spectrum of 1-Cyc1ohccy1-h-( 2,h-diohlorobeney1)- Suininotetraeoline Hydrochloridoeeoeeeeeeeeeeeeeeeeeeeeeeee 6h S. Infra-red Spectrum of 1-Cyclohay1-h-(3,hodichlorobeney1)- Sciminotetrazoline HYdPOCthridaoeeeeeeeeeeeeeeeeeeeeeeeeee 6S 6. Infra-red Spectrum of LCycbhml-h-m—mtrobenzyI-So " ininotetrezoline Hydrochloride............................. 66 7. Infra-red Spectrum of 1-Cycloheacy1uethyl-h-beney1-5-inino- $9tr83°11n0 Hydrochloride....e............................. 67 8. Infra-red Spectnm of 1-Cyuloheaq‘1methyloh-p-chlorobenzy1- 5-in1notetrezoline HYdrochloridQeeeeeeeeeeeeeeeeeeeeeeeeeee 68 9 . Infrared Spectm of 1-Cyolohexy1methy1—h4 2 ,h-dichloro- bW1)-5-w0tetraz0m0 Wcmflde....g..... Q........ 69 1.0. Infra-red Spectrum of 1-Cyclohexylmethy1-h-(3 ,h-dichloreo beneyl)oSaininotetrazoline Hydrochloride................... 70 11. Infra—red Spectrum of 1-Cyclohexylmethy1—h-munitrobmey1-S. ' iminotetreeoline Hydrochloride...........................o. 71 12 . Infre-red Spectrum of 1nCyclnheacy1metrvl-h-benzy1-S-Mnoe totreeoline..............c...............u......ac.o¢...ooo 72 13 . Infra-red Spectrum of luCyclohezylnethyleh-p-chlorobemyl- 5-1minotetrezoline....................o....oo........oo..o. 73 1h. Infre-red Spectre: of 1-Cyclohmcy1methyl-h42 ,h—dichloroo {b.n‘yl)‘5‘1N1n0t9tr35011n3eeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee 7h 15 . Infra-red Epoch-um of 1-Cyclohexy1methy1-h~(3 ,hodiohloro- benayl)~5-1n1notatreeoline..u......o........o.............. 75 16 . Infra-red Spectrum of lwycbheaqhethylah-m-nitmbenzyle S‘ininOtetr‘3011n.eeeeeeoeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee 76 X FOREWORD M The original intent of thie work us to may the emanation of h-emino—l,2,h~triezolee, however, during the course of thie inveetic getion the euthor developed e sensitivity to one or more of the conpmmde being used. This eeneitivity manifested itself in the form or ekin eruptions and made further work on this problem impossible. Part I of thie theeie ie concerned with the rceulte of the investi» gation obtained up to the time of the development of the eeneitivity. Since work on h-enino-l,2,hotrienolee m impossible , the object or the problem was changed to include e etudy of the mqlntion of l-cyoloelkyl-S-eminotetrazolee with honey]. chloride, ring eubetituted bonny]. chlorides , bgtg-phenyletlwl bromide and w—phervlpropyl bromide. The resulte of this investigation are contained in Part II of thie theeie. PART I INTRODUCTION Recent etudies of the structure of the products resulting from the flkyletion of l-sflqu-minotetrmles with elkyl halides and alkyl benzenesulfonetcs (1,2) have shown that elkyletion of s nitrogen eton or the tetruole ring occurs in preference to the expected leqlntion of the amino nitrOgen. NB, NH .111 I u c R-H/ \n 4. R'X-—----b mil/\Y-h' Na" 31—}! A report on the nethyletion of 2W1-5-uinotetruole has been made (3) in which the authors showed that the product was not 2-methy1- Smthyleninotetruole and proposed thet the product was s. nose-ionic cowonnd 1,3-dinetlvl-5-ini1wtetruele. In view of these findings e etudy of the slkylstion of other amino substituted heterocyclie systems in Ihich the heterocyclic ring contains nitrogen would be or interest. The pupa-aim of Ivonne-1,2 ,hdtriuoles utmich hsve e substituent on the amino group, has been sccomplished only in the case at 3,5-di- phewl-h-phenylsnino-l,2 ,h-trienole and compounds of this type with e substituent in the benzene ring of the phenyl group (h,5,6). The synthesis as accomplished by heating e mixture of dibemhydrazidyl dichloride (COHGCCl-H-NuCClCBHG) with phenynuydresine. This method of eynthesis does not appear to be readily applicable to the preparation or h-alkylmnino-l ,2 ,h-triasoles with alkyl groups in the three and five positions. Thus an investigation into the possibility of the introduction of an alkyl substituent onto the ammo group of Martino-1,2 ,h-triaeoles also appears to be attractive. Studies or the alkylation of h-Mno-lfi ,h-triazoles have been reported in only two instances. £1th 33; 5;. (1,8) had treated hem-1,2,h-triasole and 3, hyl-h-anino-lfi, etriasole with netlvl iodide and obtained products which they described as methiodides but did not further characterise. At the time of their studies, h-anino- 1,2,hotriasoles were thought to be dilvdro-l,2,h,5ctetrasines so that most or their conclusions were of little value. The purpose of this work is to investigate the alkylation of h-amincelfl ,h-triazolee. In addition to those reasons already stated, h-anino-l,2,h—triazoles were selected for study because they are relativer easy to prepare (9) and are usually quite stable compounds. Both the nethylation and bensylation of 3 ,S-dimethyl- and 3 , Sediphenyl. h-aninotriasole are described in this work. Development or a smitivity to the compounds handled during the course of this work halted the investigation before the objective had been reached; homer, some conclusions as to the nature or this alkylation could be realised from the work that had been done. DISCUSSION When a mixture of 3 ,5-dinothyl~h-a1d.no~l,2,h~triasole and bensyl chloride was heated at about 125°C. an exothermic reaction began which first caused the formation of a homogeneous melt then solidification of the melt. Purification of this benaylation product by recrystallisa- tion gave a somermat hygroscopic white crystalline compound. This product as very soluble in tutor and in hot alcohol but insoluble in other ccmuon organic solvents. Elemental analysis indicated that mne- benaylation had occurred to give a product sith an swirical formula of 0133150111, which, for discussion purposes, has been designated oocpouni A. The dissolution of comound A in 2H aqueous potassiun Wdroxide gave a yellow solution which upon heating on a steam bath for about one hour gave a yellos oil. Upon cooling the oil solidified to a mass of yellos crystals that, after purification by recrystallisation, gave a colorless crystalline product melting at 153-15130. The same product was obtained if the basic solution was allosed to stand for one day at room tenperature. This product was soluble in chloroform, hot nter and hot alcohol. Elemental analysis showed that the product had an enpirical formula of Gullah, and that it differed from compound 1 by loss of the elments of hydrogen chloride. This product has been designted compound 3. Treatment of an alcoholic solution of compound B with kwdrochloric acid caused the formation of a solid material which melted at 228-229°c. This product was soluble in water and hot alcohol but insoluble in other common organic solvents. The product, designated compound 0, has an emirical formla CuHuCllI‘ as sheen from elemental analysis. Thus the farmla corresponds to the addition of hydrogen chloride to cowound B and shoes'that compound 0 is isomeric with compound A. Addition of base to a solution of mound C in Inter caused the mediate formation of a powdery precipitate of compound 3. This behavior indicated that compound 0 is actually the hydrochloride of coupcund B. then compound B was nixed sith bensyl chloride and heated to about 123°C. an exothermic reaction took place. it the bogiming of the reaction a homogeneous melt developed but as the reaction proceeded the melt slowly solidified. After purification by recrystallisation a va-y Mgroscopic colorless solid was obtained ehich nelted at 171- 172°c . The material was very soluble in atom- and alcohol but quite insoluble in other organic solvents. iron the results of an elemental analysis it use deduced that the compound resulted from the intro- duction of a second bensyl group into compound B and that the coupmd had an anpirical formla of 01.11.5113,. This conpmmd has been denig- nated compound D. Treatment of compound D sith aqueous potassium hydroxide gave a yellow solution. After heating the solution on a steam bath for one hour an orange oil separated. The oil did not crystalline and did not meet to be soluble in aqueous hydrochloric acid. Developmt of a sensitivity forced a halt to further attempts to purify and commie. this product. The bencylation of 3 ,S—dimethyl-h-amino-l,2,h-triasole and the subsegient reactions of this product may be illustrated by the follow- mg scheme: H01 Gllnlscma , C N—o—Q‘ m o i u MC‘HBCH'CI KOH( ,) KOH flop. 228-229 C 033 \N/C.CH3 211515011514 G g N ‘ w C 11.43.01 §H3 mops 19141-1950‘3. “up. 153‘151‘00 G Cauofia ‘ . cuflumu‘ n.p. 19749800. mp. 171~172°c. Heating a mixture of 3,5—diphewl-h-eMno-l,2,h~triasols and mi chloride to about 125°C. initiated an exothm'nic reaction. Tron this reaction a colorless solid material was isolated that decon- posed when heated to 202—20130. Analysis of this product indicated that nonobensylation had occurred. Treatment of the diphenyl nonobensylation product sith aqueous potassium hydroxide gave a yellow solution. After heating the solution on a steam bath for about one hour a red oil had separated. when attempts were made to purify this product only tarry material use obtained. The benzylation of 3 , S—diphaayl—h-afim—lfi ,hatriasole appears to produce a compound analogous to compound 1.. In the treatsent of the benvlation product sith base the reaction appears to be similar to the reaction of the nonobensylated 3 , Sodinethyl compound sith base, at least in the first stages, in that a yellow solution, which produces an oily product upon heating or after a long period at room tempera- ture, forms in both cases. The difficulty encountered in isolating a pure product could then occur either because of the instability of the dipherxyl product or because the reaction took a differmt course during a later stage. Methylation of 3 ,S-dimethyl-n and 3,5odiphem1oh-mino-1,2 ,h—triazole by heating with methyl benseneeulfonate led to an exothermic reaction as in the bonnylation. With 3 ,S-dimetrwl-heanmzo-l,2,h-triasole a crystalline product was isolated which exhibited the correct analysis for a bensenesulfcnic acid salt of a methylated 3,5-dinefi1yl-h-aniuoe 1,2 ,h-triasole. With 3 , Sodiphexwl-h-aMno-lfi ,h-triasole , however, only a glassy material that would not crystallize could be isolated. Heating notmrlatod 3.5-dinettwl-h-uino-l,2 ,hrtriasole with 2N potassium hydroxide did not appear to cause any reaction but ehen the base strength ens increased to w a yellouP color developed and a small amount of precipitate formed. The behavior of the precipitate upon heating indicated that it night be an organic salt. With the hope of synthesizing an authentic sample of 3, Sodinethylu h-ddnothylandno-l,2,h~triasole, 3,5-dinethyl-h-amino-l,2,h-triasole use treated with a moire of formic acid and formaldehyde according to the Eschweiler-Clarke procedure (10). The prochict obtained fron this treatncnt could not be crystallised either as a free base or as s mdrochloride. Although this method of reductive alkylation is generally applicwle to the preparation of dimcthylated amines , it has been reported to be unsuccessful in instances shore the amino group is influenced by a strongly electronegative group (10) or where the amine reacts with formaldehyde to give an unreactive condensation product (11) . The infra-red absorption spectra of 3 ,S-dinethyl-h-aminodfl ,h- triasole, compound A, compound B, compound 0 and compound D were obtained in order to aid in the characterization of these compounds. Inspection of these spectra, sham in Figure 1, reveals appreciable differences in the location of the bands exhibited by these compounds. Although these differences may be indicative of profound structural differences, it met be recalled that three of the compounds probably involve ionic structures which could cause spectral differences while still retaining considerable structural cindlarity. bromination of the individual spectra, however, allows some conclusions to be dram as to the nature of the structures of the compounds. The spectrum of compound a shows two notable features which give some insight as to the structure of this compound. Absorption bands at 3.16, 3.26 and 6.08 )1 may be attributed to the presence of a primer] shrine function (12) while the absence of any absorption at 33544.0 p indicates the absence of an amine hydrochloride structure (13). rm, one may conclude that compound a is probably a quaternary chloride and not a hydrochloride. Compound B shows a single absorption band at 3.16 p and no absorp- tion near 6.1 p, indicating that it has a secondary amine function and not a primry amino m. The strong absorption bands at 14.0 p and at 5.61 )1 exhibited in the spectrum of cowound 0 clearly demonstrate that compound 0 is an Figure 1. Infra-red Spectra . 1. 3 ,S-Dinethy1~h-anino~l,2, h-triasole 2. Corpound A ‘ 3. Compound B ' 1;. Compound 0 h S. Commund D Per Cent Transmission 80 6O 40 2O 80 6O 40 20 80 6O 40 20 80 60 40 20 80 60 no 20 : 1*“. In ,flPWfi r’”“‘n ./ /‘rfl\ V f m 'v V [1/ VW V 1W] V WWW/“x M, 'v x V M/‘AA. urn/mfg / NW NE U\A v V \F u] it 1 u v A f A U M 1 'W U \ v 6 8 10 12 Wavelength (microns) 14 amine hydrochloride (13). This coupled with the fact that compound C is formed by addition of hydrochloric acid to the free base B and that compound B is rapidly regenerated when compound 0 is treated with cold aqueous potassium hydrmdde solution, shows that compound C is simply the hydrochloride of compound B. The most notable feature of the spectrum of compound D is its lack of absorption in the four micron region which indicates that this compound is not an amine hydrochloride. There is also a marked similarity in the spectrum of compound I) and that of compound A in the region beyond 8 )1. This may be taken as indicative of a similar ring structure since the region of the infra-red spectrum at wavelengths longer than 8 )1 is associated with vibrations of the skeletal structure as a whole. Attempts sore made to remove the benzyl pcup from compound A and from compound B by catalytic hydrogenolyais using the procedure described by Birkhcfer (1h). Compound B resisted all attempts at catalytic debenzylation uhile compound 4 took up the theoretical amount of hydrogen in six hours under the same conditions. The debensylation of compound A gave a strong odor of toluene in the reaction bottle but the product isolated from the reaction mixture did not are a sharp nelting point after several recrystallisation. The development of a sensitivity to the compounds handled in this work halted further attempts to purify this material. In the Scaminotetrasole series Garbrecht and Herbst (15,16) have reported that a bensyl group attached to the one position of the 10 tetrasole ring can be removed in a few hours by catalytic hydro— gsuolysis according to the procedure given by Birkhofer wile a bensyl poup attachedto theamino groupcanbe remved onlyarter long treatment, it at all. This behavior may be analogous to the situation with cow 1 and compound B in um the bensyl group may be attached to a heterocyclic ring nitrOgen atom in compound 1 and attached to an saline group nitrogen atom in compound B, thus accounting for the relatively easy removal of the bensyl group from compound A and the lack of reactivity of compound B. Treatment of h-amino«-l,2,h-triasoles with nitrous acid has been reported to rsmve the amino group with the resulting formation of a 1,8,hotrissols and nitrous oxide according to the follosing reaction scheme (1,7,18,19,20): N—O—fl 3*}! II II II cu,-!\ /c-ca, 9 mm, ---> 0334\3/0-0113 9 mo '6 3,0 I 3 B 5—2: In the hope of obtaining a similar reaction at the primary amino group, compound A was treated with nitrous acid. A vigorous evolution or a colorless gas occurred but in an attempt to isolate the organic product only an oil that darkened upon heating was obtained. Attempts to purify the product sec-s halted because of the development or a sensitivity to scans of these «wounds. Treatnmrt of compound B with nitrous acid in s like manner resulted in the fomtion of a blue color but no gas evolution was observed. It was thought that the reaction of compound 1 with nitrous acid might establish the structure of compound A, since if the honey]. group of compound 1 were located on a nitrogen atom of a 1,2,h-triazole ring the product of this reaction should be 3 , Methyl-1,2 ,h-triacole with a bensyl substituent in either the one or tour position. The reaction does serve as a qualitative test for a primary, secondary, or tertim amino group since a primary amino group would evolve nitrous oxide, a secondary amino group ehould form a N-nitroso derivative that would be colored, while a tertiary amino pomp would not react with nitrous acid. is a walltative test the reaction shows that compound A has a primary amino group and that compmmd B has a secondary amino group. From the reactions and infirm-spectrum of compound A one may con- clude that the material is a quaternary chloride with a primary andno group and probably contains a bonny]. group attached to a nitrogen atom of a heterocyclic ring. These facts may be explained by strocwree or the type I and II. 9 n—o—n n—--— 4333436116 011,-! e/g-Cfla Cl- CHa-C\H/C-CH3 Cl. NH. \CHrCeHS I'm. I II A compound of the type shoun in I, arising from benzylation of the h-position of the triazole ring, presents a situation similar to that arising when unsym. dieubetituted hydrazinee are treated with an alkflating agent to give a hydrazinium salt. In the benzylation of Nobenzyl-Nophenylhydrazine with banal chloride N ,N-dibenzyl-N-phenyl- hyflruinium chloride has been shown to arise (21). A structure of the type shoun in II, arising from bensylation of the l—position of the triazole ring, would be expected to be stabilized by virture of the hybrid arising from resonance contributions of forms, II, III and IV, of the cations. O fi—-—-fi-CH,CBH5 H—o—H-CHr-CGHB n-v—u-carcsna ...... J *r CH:'°\ /C‘CHa CH3 \ 9 ~03; ens-0\choH3 ’i‘ '3' ' r NH, NH2 NH2 II III IV The infra-red spectrum and properties of compound B indicate that this material is a heterocyclic compound with a secondary amine inaction. From a consideration of the reactions which structures of the type I and II are likely to undergo upon treatment with base, two structures for eowound B can be visualized as arising from the reactions illus— tratsd in the relieving acme: N—u-—N CH3-0\ /C-CH3 la OH" x - V II n 01" CI‘I;,--C:\o/O«-CH3 H / \ 11,11 0113-0ch. 3......)m 3—43 OH II I I Celia-CH, NH. —-—N I CBHBCH. VI VII a. on 3 ens-c c-caa '“'“ ’ (ma-c ’ “'”’ \E/ \lf)\0H “Ha - NHa Cl II VIII [CH’CeHa Ill-o—N-CH.C°H5 s—-— \ ° \n-o—u NH\ H NH: 2 IX X In scheme la the product would be, 3 ,S-dimethyl-h-benzylnmnml,2,h- triazole and could result from the migration of the benzyl group in a manner somewhat similar to that of a Stevens rearrangement or from elimination of the honey]. group as a bensyl carbonium ion followed by attack of the mine nitrogen by the benzyl carboniuxn ion. This product would also serve to explain the difficulty encountered in attempts to remove the benzyl group by hydrogenolyeis. Either scheme lb or scheme 2 uould lead to the same product l-u-benzyl- 3,6-dimethy1-1,h-curodro-1,2,u,5-eetnzme which could be in tautoneric equilibrium with other dihydrool,2,h,S-tetrasine structures N—o—NH N—a—K H H=Iu==N // \ l y \ cu,-c\ i411, -—--- ca,-o\ /c-cn, «- / [0-0113 ’3‘” Irv“ ”WM 03,031, 03.0.3115 03.0335 I XI III In scheme lb compound A is converted by base into thebydrssidine VI which could then cyclize as shown, shile in scheme 2' the fwdrazidine II, which could arise from a pseudo base such as VIII, is cyclized to give the same product as that resulting from scheme lb. 01’ the two products which would result from the above schemes 3,5—dimet1'qu-h-benzthdno-lfi ,h~triasole seems more probable since eonpound B is a colorless solid that is stable towards heat while the dihydro-l,2,h,S—vtetrasines are reported to be yellow solids that rearrange to h-amino-l,2,h-triasoles upon heating (22). The behavior of compound D could also be explained since the benzylation of 3,S—dimethyl-h-bmzylandno-l,2,h-triasole night be expected to give a product similar to compound A. 15 n--n n—-—u - ll Bella-0:130; II II 03- 011;,.,--c\n/c-cu3 (3113.05. /c--<:H3 I \ V XIII This would lead to a quaternary chloride and would account for the ammun- 5.. the spectra a: comnd A and eonpound D. since hum-1,2,h-triasoles have been reported to condense with alderwdes and ketones to give a hydrasone-like product (23,210, it was felt that this might offer an opportunity to synthesise 3, 5-6130th- hobsnsylanino-l,2,h-triasole through catalytic hydrogenation or the condensation product of bemaldekwde and 3,5-dinetm1-hqnim-l,2,h~ $118801. 0 CH CH CH N s a" 3 H C, 3 H s 0/ 3 . \ | :N-NH. w. | :Mx-cncana .333. | lfi-NH-CH-CGHS u-c and Nod (EH3 on, cu3 When bonsaldervde and 3 ,Sadinetlvl—h-am-l,2,h~trieaole were heated tagether a yellow oil was the only product isolated. Condensation of anisaldehyde sith 3 ,Sodiuethyl-h-andno-l,2,h-triasole also gave a yellow oil, but salicylaldehyde and 3 ,Sedimethyl-h-aminodfl ,lr-triasole condmsed to produce a small mount of a crystalline product. Hydrogenation of the bensaldehyde condensation product resulted in a rapid initial uptake of ivdrogen uhich quickly slowed but did not stop after 1 mole of hydrogen had been taken up. stopping the reaction 16 after absorption or various percentages of the calculated ludrogen uptake gave only an unstable oil and some 3 , 5-dimethyl-heami.no- 1,2,h-triasole. The condensation of bonsaldelwde with 3 ,S-diphenylah-enino- 1,2,hdtrissole gave an oily product; however, h-snino-l,2,h~trissole and benealdehyde condensed rapidly to give a solid product. when the condmsation product of bonsaldehyde and hvaninoolfi ,h-triascls as subjected to the some hydrogenation conditions as the condensation product of bensaldemrde and 3 ,S-disethyl-h-amino-l,2 ,h-triaaole no reaction occurred. Increasing the taperature and wont of catalyst also failed to induce a reaction. 17 EXPERIMENTAL Preparation of 3 ,S-Disubstituted bimbo-1,2 ,h—‘rriasoles The preparation of 3 ,5-dinethyl—h-ami.no-l,2,h-triasole and 3,5di- pheznrlv-h-nminoml,,I:~ ,hotriazole was accenplished by the method reported by Garrison and Herbst (9). Interaction o: dibenzoyl hydrazine and hydrazine pave the diphenyl command while the dinethyl compound me synthesized from acetic acid and hydrazine . -Di - -amino~l 2 ctriasole A mixture of 21; g. (0.10 mole) of dibensayl Ivdrssine, prepared according to Hett (25). and 11.8 g. (0.20 sole) or an 85% mun-duo bursts solution was sealed in a Pyrex comustion tube. The tube us heated to 185°C. odd maintaixwd at that taupe-dour. for two doors. After Opening the tube a shite solid material as obtained that gradually darkened upon standing. The contents of the tube were dissolved in boiling ethanol and digested with Norite. Filtration and cooling produced 12.? 3. (51d based on the dibensoyl hydrazine) of lusteroue white platelets of 3,5odiphemrlch-anino-lfl ,h-trissole melting at 261-262°C. ' ‘hvo preparations were carried out as described above withthe “institution of anrydrous hydrasine tor the 8 5% hydrasine hydrate solution. In both instances successive pressures were developed in the sealed tubes as noted upon opening the tubes. 1 very strong odor of 18 amonia was also noted upon opening the tubes. The yields of 3,5. diphexvl~h~amino~l,2,h-triazole from these runs were 143% and thZ. id-Mdld-mhemm a round-bottomed flask nus charged with 2&0 g. (h moles) of glacial acetic acid and cooled in an ice bath. With continued cooling, 355 g. (6 miss) of an 85% hydrazino fwdrste solution was added draperies at a rate idlich kept the temperature of the reaction mixture below 50°C. After couplets addition of the hydrazine solution the flask m fitted for distillation and slowly heated on an oil bath to 225°C. The tanperature was then maintained at this level for six hours. Dining the heating period the course of the reaction ms follomd by measuring the water and hydrazine which was distilled from the reaction vessel. At the and of the heating period the flask contained a colorless liquid which crystallized upon cooling. Recrystallization or the product from isopropyl alcohol gave a crop of thick prisms. Concentration of the mther liquor to about half its former volume gave a second crop of thick prisms. The total yield of 3 ,5odimethyla-h-amino-l,2,h-triecole melting at 197-19800. was 159 g. (71% based on the acetic acid). A small amount or 3,5-dimethy? -h-udno-l,2,h—triazole was dissolved in 25 ml. of ethanol and acidified nth concentrated hydrochloric acid solution. Addition of 25 ml. of other caused the slow formation or fine needlealilce crystals. Recmullisation from 50% ieOpropyl ether- ethanol gave needles of 3 ,S-dinetlvl-h-andnov-l,2 ,hutriazole hydrc- _ chloride melting at fill-229°C. (19). l9 Alkylation of 3 ,S-Dimbetitntod h-MnOol,2,h-Triazolee Both 3 ,Sodiphemyl- end 3 ,S—dimethyl-h-cmino-l,2 ,h-trmole were methylated and benzylnted by simply heating a mixture of the Ironmo- 1,2 ,h-triazole with benzyl chloride or methyl bonze‘noeulfonate. An exothermic reaction occurred after which the product was purified by recrystallization. Metrilation of 2.&2hgl-h-andno«rl I2 ,gmle A mixture consisting of 9.1;}; g. (0.01.; mole) of 3,5—dipherw1—ho Wal,2,h~msole and 7.25 g. (0.0M mole) of methyl benzeneonli‘cmte In. heated on an oil bath. Wham the temperature or the mixture ruched chant 70°C., on exothermic reaction began which raced the towereture to 120°C. and wood the formation of a homogeneous liquid. After the ruction had subsided come, the mixture In: maintained at 110°C. for 15 mantel. The reaction mixture In dimlved in boiling leoprcpyl alcohol and upon cooling 3 yellow viscoue oil separated. Several attempt: to crystalline the oily Intern]. by dimlving in hot leoprppyl elcohol end tub-ewent cooling gave only the lame 718G011. oil. the oil Ill then allowed to Itend in e um deliccator over phosphorus: pentoxide for three woke but at the end of that period the product had only thickened slightly. It was not Wed farther. Met lotion of t l- «amino-1 2 -triazolo A mixture of 5.60 g. (0.05 mole) of 3,5-dimot1vl:h~amd.no-l,2,h~ triuole and 8.91; g. (0.052 mole) of methyl benzeneeulfonate m Ilovly 20 hosted on an oil bath. is the temperature of the mixture reached 50°C. a vigorous exothermic reaction began which caused the temperature of the mixture to reach 125°C. During the course of the reaction the mixture became a homogeneous yellow liquid. After the exothermic reaction had subsided the contents or the reaction vessel were main- tained at 90°C. for 15 umtes. Digestion 01' the material um a small amount of boiling isopropyl alcohol produced a white powdery precipitate. Recrystallization from 90% isopr0py1 alcohol produced a fine white precipitate. Drying the precipitate for five days in a vacuum desiccator over phosphorous pentoxide followed by eight hours drying in an oven at 130°C. gave 11.2 g. (79% yield based on 3 ,S-di- neflxyl-h—Mno-l,2,h-triasole and scanning a 1:1 reaction ratio) of a product melting at 189-190°c . ' Analysis: Calculated for c, Hmfl‘oasa C, 116.5%! H, 5.7%) x, 19.7%: a, 11.0%. Fame: 0. 1.65%: H. 5.6%: H, 19.7%: a, 11.0%. Emlation or 3 . 5-dimethyl-y-amino-l. 2 Igmtriazsole A mixture of 11.2 g. (0.10 mole) of 3,5-dimethyl-Mnoal,2,h~ trieuols and 11..o g. (0.11 mole) o: bensyl cm... was slowly heated on an oil bath. is the temperature reached 10500. a homogeneous melt formed and at 125°C. an exothermic reaction began which carried the temperature to 180°C. During the course of the uothermic reaction the melt gadually solidified. The product was kept at 11.0%. for half an hour after the reaction mixture began to cool. The solid product was dissolved in hot isopropyl alcohol and upon cooling a fine 21 white precipitate formed. From a second recrystallisation from isopropyl alcohol small platelets were obtained that were dried in a vacuum desiccator and in an oven at 110°C. The yield of compound A was 19.5 g. (821 based upon 3,5-dinethyl-h-anino-1,2,h-triazols and a 1:1 reaction ratio), melting u. 191.-195°C . Amlysisl Calculated for C, H1501)!“ C, 55.3%; H, 6.355; 01, 11.3%; N. 233%. Found: c, 55.9%; H, 6.3% 01, 11.3%) N. 23.0%. In four repetitions of the above procedure the yield of compound A was found to range from 80% to 90%. Reaction of Monoalkylated Products with Base Treatment of mflkwggted gig-dimetml-h-amino-l |2 .hutriazole with base A solution containing 5.? g. (0.02 mole) of the product of the reaction of 3, Soonethyl-h-amino-l ,2 ,hutriasole with methyl benzene. sultonate in 50 ul. of 2N potassium mdrozide solution was heated on a steam bath for one hour. No evidence of reaction was observed after the solution was cooled, so 5.6 g. (0.10 mole) of potassium Ivdroxide us dissolved in the solution and the solution again heated on a steam bath. After about 15 nimtcs a bright yellow color developed which gradually faded. The color had almost completely disappeared after one hour of heating, when the solution was removed and cooled. A null amount or powdery precipitate which had formed upon cooling was removed and recrystallised from isopropyl alcohol containing about 35$ isopropyl other. This product did not melt when heated to 273°C. and shared slightly when heated in a direct flame. 22 nt otbe lat , h 1- -anino-l2 otmmle thbase A solution consisting of 7.21; 3. (0.020 mole) of the product result- in; from treatnmt of 3 .S-diphexwl-h-enino-lfi ,h-triasele with benayl chloride, 1.7 g. (0.03 mole) of potassium marcxide and 50 ml. of water use gently samd on a steam bath. After a for dates the solution turned bright yellow and after one hour a dark red oil had separated. The solution us then cooled and the oil extracted sith chloroform. Gradual addition or isopropyl ether to the hot chloroform attracts until a faint turbidity sea observed followed by ales cooling of the solution again caused separation or a red oil. The oil use separated tron the solvent and heated with bensene. This treatment resulted in the forution of a black tar. In a second reaction using the above procedure the chloroform solution of the red oil sea treated with dry hydrogen chloride. A yellow-orange send-solid material precipitated and was separated. The resulting product ins extracted 1with dilute hydrochloric acid but neutralization of the acid extracts failed to give an unsoluble product. TreatxeenLgf benqlcted 2, goddmeggl-hewl ,2 ,3 otriasole 5th base Dissolving 19.0 g. (0.08 mole) of compound A, the product of the reaction of 3,5—dimethy1-h-anino~l,2,h~triazcle with banal chloride, in 100 ml. or 2 H potassium hydroxide solution gave a colorless solution mioh slowly tin-nod yellow upon standing. Gentle heating of the solu- tion on a steam bath for about one hour caused the formation of a white precipitate and a yellow-orange oil. Upon cooling, the oil crystallised to a mass of pale yellow crystals. If the alkaline solution see not 23 heated on a steam bath, but allowed to stand at room temperature for one dc the same pale yellow precipitate was famed. The solid material was removed by filtration and thoroughly washed with outer. Purifi- cation of the crude product by recrystallisation first from water, than from isoprOpyl alcohol containing 20% isopropyl other gave 13.0 g. (80% based on the assumption that the elements of hydrogen chloride sore eliminated by this treatment) of fine, colorless needles melting at 153-15h°0. This compound was designated compound B. Analysis: Calculated for sunny“ 0, 65.3%: H, 7.0%; on, 27.73. Found: C, 65.6%: H, 7.1%) H, 27.8%. a small portion of the base, compound B, was dissolved in ethanol then acidified sith concentrated hydrochloric acid. Addition of a small amount of ether, initiated the formation of fine needle-dike crystals. Recrystallisaticn of this product from isopropyl alcohol containing 20% isopropyl ether gave a command, designated compound 0, melting do arr-228%. Analysis: Calculated for c,,n,,cm,: c, 55.3%: H. 6.3%: 03-, 1h.9%3 N, 2305%' Found! C. 55.0%: H, 6.3%; Ci, 15.0%; H. 23.1%. When a small portion of cowound C was dissolved in water and made alkaline ulth potassinm hydroxide solution, a powdery solid was obtained. Recrystallisation of the solid material from water gave tine needles of the base, compound B, melting at 153-15130. No depression in the nelting point was observed when this product was mixed with an authentic sample of compound B. 2h Benzylation of Compound 3 a mixture of 7.1h g. (0.030 mole) of the base, compound B, obtained by'benzylation of 3,Sodimethyl-h-amino-l,2,h-triazole, and 5.06 g. (0.0h0 mole) of benzyl chloride was slowly heated on an oil bath. A homogeneous melt formed when the temperature of the reaction mixture reached 115°C. and when the temperature reached 125°C. an axothernic reaction‘began that raised the temperature to thOC. The temperature of the melt was then slowly raised to 170°C. and held there for one hour. Upon.cooling the melt crystallized and was dissolved invboiling inaprOpyl alcohol. The pale orange solution was digested Iith Norite and filtered thus giving an almost colorless solution. Addition of ineprOpyl other until the solution contained about 50% other initiated the growth of fine cryotale. The product was removed and recrystallized fron.a 50% isopropyl aloohol-ieopropyl other solution giving 8.h z. (77% based upon compound B and the assumption that the reaction ratio was 131) of very fine, colorless needles melting at 171-172°c . This product was designated as compound I: . Analysis: Calculated for 0183,101N,: C, 65.8%; H, 6.h%; 01’ 10.8%, H. 17.0%. Found: c, 65.6%; H, 6.7%; Ci, 10.95%; 1:, 16.8%. Reaction of Compound D with Base The addition of 6.6 g. (0.020 mole) of compound D to SO-ml. of 2 N potassiun.hydroxide solution gave a.yellow'eolution.uhich slowly‘became turbid. Heating on a steam'bath for one hour resulted in the separation 25 of I. yellow oil. The mixture was diluted with 150 m1. of at... cooled and extracted with chloroform. Evaporation of the chloroform left an oily residue which as dissolved in impropyl alcohol and acidified Iith concentrated hydrochloric acid solution. The addition of other to the alcoholic solution. did not can” the formation of any precipitate. Removal of the solvents left a dark red oil which was digested tith benzene for one hour thm allosed to stand, uith occasional stirring, for three dm. it the end of this treatment some color us observed in the became but the oil did not appear to have changed. Hydrogenolysia of Bensylated Products Attempts to rows the bensyl group from the benaylated products vars carried out in the manner recomended by Birkofer (1).) for catalytic debenvlation. The reaction was carried out in a low pressure mates using palladium as a catalyst and either alcohol or acetic acid as the solvent. Atmted «Elation of 93m 3 A Parr bottle was charged sith h.3 g. (0.02 M10) of the base obtained from the product of the reaction of 3,5-dinettvl—h-anim- 1,2,hdtriaaole and bonsyl chloride, compound B, 2.5 ml. of 12 B hydro- chloric acid solution, 5 g. of 51 palladium on charcoal and 160 al. of 90$ ethanol. The contents of the flask uere put under a ivdrogen pressure of 1.5 p.s.i. and shaken for 25 hours. During the last ten hours of shaking the bottle was heated to 65°C. and held at that tauperature. At the end of this period no pressure drop had been 26 observed (the theoretical pressure drop was 2 lbs.) , so the hydrOgen us released. No odor of toluene could be detected in the bottle. After removal of the catalyst and addition or 100 ml. of other to the alcoholic solution a precipitate of the hydrochloride of compound 8, compound 0, was obtained. Treatment of this hydrochloride with aqueous potassium hydroxide gave the starting material, compound B. Other attempts to debennylate compound B were made using the procedure described above but with either omdssion or the hydrochloric acid or sith the use of glacial acetic acid as the solvent. In no instance see any pressure drop observed or any toluene odor detected. The only product isolated use the starting material or its hydrochloride. A check or the activity of the 52 palladima on charcoal catalyst sue made by twdrogenslysis of a 0.020 mole sample of l-bennyl-S-acetyla udmtetrasole. This compound gave the calculated pressure drop in 20 sdmtes and upon opening the bottle a very strong odor of toluene no detected. wage oLcogmund A A Parr bottle as charged with 11.9 g. (0.050 mole) of the product of the reaction of 3 , 5-ddnethyl-h—enincol,2,hotriasole and benzyl chloride, compound A] 5 g. of 5% palladium on charcoal and 100 ml. of 951 ethanol. Shaking under an initial hydrogen pressure of 1.7.0 p.s.i. at 66°C. for six hours resulted in a pressure drop of 3.8 lbs. (calculated drop: 13.0 lbs.). Upon opening of the bottle a strong odor of toluene us detectw. The solution was concentrated to about 50 ml. then acidified with concentrated hydrochloric acid. Addition of a small 27 amount of ether induced the formation of a powdery shite precipitate. The precipitate nelted over a range of 25° beginning at 190°C. and after four recrystallizations from 50% ieopropyl alcohol-ieoprOpyl other a melting range of 216-.12225‘D C. was observed. Condensation of h-aminoalfi ,h-triaeoles sith Aldehydcs The condensation of aromatic aldehvdes Idth butane-1,2 ,h—triasoles us conducted according to the procedures described by Ruhenann and Karim (23) and by Boles and Weber (2h). Condensation sac effected by heating ewinolar amounts or the aldehyde and the h-ani.no-l,2,h- triasole in an ethamlico solution containing a catalytic amount of piperidine or acetic acid. Metbylation Ins attempted by the reaction of 3 ,S-dinethylohc-emino- 1,2,h-triazole with formic acid and formaldehyde according to the Eschweiler-Clarke method (10). The reaction appeared to proceed some- that slower than 1- normal and no crystalline product could be isolated. Cond otion or aromatic ald es with ' 1- «nine-1 2 - so i nature of about 0.2 g. of 3,5~dinethyl-h-andno-l,2,h-triasele and about 0.2 ml. or salicylaldehyde ens dissolved in ethanol and 2 drops of piperidine added. The solution turned pale yellos upon gentle nrflngcnastesnbath. WaddingenougheutertodilntetoSm ethanolthesolnticnusallcsedtostandtoreneday. ittheendot this tine a feathery shite precipitate had formed . The precipitate was recrystallised 50% aqueous ethanol and gave a product melting at 28 189-19000. (23). Using the procedure Just described with the sub- stitution of 2 drops of acetic acid for the piperidine the same product as obtained. . Attempts were made to condense bonsaldehyde and anisaldehyde with 3 ,5-dinettvl-h-mino-lfi ,h-triaaole using the above procedure, with both acetic acid and piperidine catalyst. In all instances a yellow color was observed but no crystalline product was obtained. The use of this procedure with 3 ,S-diphenyl-h-amino-l,2 ,h-triasole and bensaldelvde also failed to give an solid product. condensation)! benzgldgxzde with g—Mo-1,2,g-tr1aao1c a nature of 16.8 g. (0.20 mole) of h—Mnc-l,2,h-triasole and 21.2 g. (0.20 mole) of benzaldehyde was dissolved in 75 ml. of ethanol and five drops of piperidine added to the solution. The solution was heated gently on a steam bath for one hour and then cooled. A mass of fluffy white crystals had famed and this product was recrystallized from ethanol. The yield of product, previous]; described as H-bmsylideneo h—aflm-l,2,h-triazole (23). melting at ins-172%. was 26.1 c. (78%). R on of -din l-h~amd.no-l 2 -triazole uith formic acid- 0 e To 22.]; g. (0.2 mole) of 3,5-dimethyl-lrmol,2,h~triaaole, 51.0 g. (1 mole) of 901 formic acid was slowly added with cooling. After addition or 37.6 3. (Odd; mole) of a 35% aqueous formaldehyde solution the mixture use heated on a steam bath for 30 hours. During the early part of the heating period evolution or a gas was observed but as heating was continued the evolution became unnoticeable. it the 29 end of the heating period 100 ml. of 3 H hydrochloric acid was added to the reaction fixture and heating continued for five hows. Evaporation of the excess liquids under reduced pressure gave a yellos viscous oil. The oil to. readily taken up in isOprcpyl alcohol but addition of ether caused the separation of the oil. After standing under bensene for several days the oil appeared to be unchanged. Hydrogenation of Bensaldehyde-h-Amino~l,2 ,h- Triasole hixtures A mixture of bensaldehyde and 3,5-dinethyl-h-enino-l,2,h-triaaole, which had been heatedtogether, was. subjected to hydrogenation using platinum oxide catalyst. The mixture took up hydrogen but the reaction did not appear to involve a one mole addition. The only products isolated were starting material and unstable oils. magmatic]. of benoaldghEe-QIMl-g-m-lI2IQ-triasole A mixture of 5.6 g. (0.05 mole) of 3,5—dinethyl-hceninocl,2,h- triasole and 5.3 g. (0.050 mole) of benssldehyde was heated together on a steam bathuntil a thick oil had been produced. This product was transferred to a Parr bottle along with 100 ml. of 953 ethanol and 0.2 g. of. platimn oxide. The bottle uas put under a hydrogen pressure of 1.7.5 p.s.i. and shaking begun. After 30 minutes a drop of 2.0 lbs. had occurred (h.0 lbs. theoretical), after seven hours a h.0 lb. drop was observed and after eleven hours a five lb. drop was noted. The bottle was opened at this time and no odor of unnonia or toluene noted. Removal of the catalyst and evaporation or most of the ethanol under 30 ' reduced presmre gave a yellow oil. The oil was taken up in chloro- tors and use regenerated by evaporation of the chloroform. During these steps the oil darkened considerably until a very dark red tar was left. In other runs using the same technique as above the reaction was stopped after various percentages of the theoretical ivdrogen drop had occurred. When the reaction see stopped before 125% of the theoretical drop had been observed a red oil sink: to that described above was obtained along with various amounts of starting material. The per cent of starting material isolated after various percentages of the theoretical pressure dmp my be summrised as follows: 60% drop- 95% or the starting material. 75% draw-50$ or the starting natcial, 100% W151 of the starting lateral. Attempted mo enation of the benzalde s h-amino-l 2 hatriasole gorflensation uc ’ A Parr bottle was charged nth 17.2 g. (0.10 mole) of the conden- sation product of bensaldehwde with h-amino-l,2,h—triasole, 150 ml. of ethanol and 0.2 g. of platinum oxide. The bottle me put under an initial hydrogen pressure of 1.7.2 p.s.i. and shaken for 15 hours. During this time the temperature was held at 66°C . and after five hours, shaking was stopped and an additional 0.2 g. of platinum oxide added to the Me. No pressure drop was observed during this 15 hour treatment. The bottle was opened and the contents filtered while hot. Cooling the filtrate gave a crop of fluffy white crystals of the starting material melting at 170-17100 . Concentration of the filtrate 31 to about 50 ml. gave ssecond crop of starting material. The total want of starting mterisl isolated m 15 .9 g. (96% of the original starting material) . Reaction of Nitrous Acid with 3 ,S-Dimethyl-h-sminc— 1,2 ,ls-triezole and Its Bensylstion Products geactiorgufivdimetglggenfino-l.2 ,E-tgfgsolg git}; 113W A solution of 7.}; g. (0.050 mole) of 3,5-dime‘thybh-emino-l,2,h- triseolo hydrochloride dissolved in 50 ml. or water was cooled in an ice both. A solution or 3.8 g. (0.055 sole) of ladies nitrite in about 15 ml. of 1meter was then slowly added to the cold triazole solution. A vigorous evolution of a. colorless gas occurred upon mixing the two solutions. After complete addition of the sodium nitrite the resulting mixture was allowed to come to room temperature, then slowly warned on a steam bath. Evaporation of the liquid under reduced pressure left a residue of white crystals. By hosting on a. steam both under reduced pressure 3.9 g. of 3 ,S-dimethyl-l,2,h-triszole melting at lh0¢lhl°0. (19) was collected by sublimation (81% yield based on 3,5-dimethyl-h~ W1,2.h-triszole) . Reaction of Command 1 with nitrous acid A solution of 5.95 g. (0.025 mole) of the product of the reaction oi‘ 3,5-dinetlwl-h~smino~l,2,hdtrissolo with bensyl chloride, comm A. and 2.5 ml. of 12 N hydrochloric acid (0.03 soles) in 2.5 ml. of water as cooled in on ice bath. Addition, in small portions, of a solution contsining 1.9 3. (0.0275 mole) of sodium nitrite dissolved in 10 ml. of 32 water produced s vigorous evolution of s colorless gas. The reaction mixture was allowed to come to room temperature then gently warned on s steam bath for half an hour. it this point the solution had token on a pale yellow color. The solution was cooled and 1.9 g. (0.031; mole) of potassium hydroxide we added. A yellow oil separated and was extracted with chloroform. Bubbling hydrogen chloride through the chloroform eoctrscts resulted in the formation of e brown-wrongs pro- cipitste. The solid we removed by filtration and dissolved in ethanol. Addition of ether gave only a dark yellow oil. Reagtion pfficompound C with nitrouggcid A well sample of compound 0 dissolved in Inter and cooled in an ice bath was treated with e sodium nitrite solution. No evolution of gas was detected and the solution turned blue. Standing for several hours at room tmporature eased the precipitation of a dark blue solid material. ' Infra-red Absorption Spectra The infra-«ed absorption spectra. of 3 ,S-‘diJnethyl-hosrdno-lfi ,1;- triszolc, caupoum A, compound B, compound 0 and compound D were cbtsined using s Perkin-Elmer Doubleheam Recording Spectrophotometer, Model 21. All samples were run as oil mine. The spectra are repro- duced in Figure l on page 8. PART II .33 INTRODUCTION Rsouxtly interest in ring substituted inimtetrasolines has developed due to their durabiologioal activity. A study by Rsutner, Peters and Elslager (26) has revealed that new isinotstraselins hydro- chlorides were active against a rather wide range of bacteria as well asssvu'alsulturssotfungiandpretasoa. Adstailedreportefthe effect of structure on the antitriohomnal activity us also wanted in this paper. i series of l-alkyloh-aralkyl-S-innotstrasolins hydrochlorides efthetypssheminlwrstutedgmagainstmmomsm by Reutner .s__t_ 5;. NH oHCl 063,4 CHa)y'li‘/c\‘f’(93a)xCHs l—N I It was found that triohosonaeidal activity reached a maximum when 1 - l and x s 7, that is, when one substituent was benzyl and the other nbstitusnt n-ootyl. Further tests revealed that substituents in the plum ring of the bansyl group had .. effect on the activity, with a .11” being reached when a 2353 chlors substituent was present in the phenyl ring. ‘mus ths greatest activity of all the compounds tested was found in lon-octyl-h-p-ohlorobemyl-S-ininotetrasolins hydrochloride. Since these remlts show that the activity of 3h 5-ind.notetrsso1ine hydrochlorides is greatly dependent upon the neture of en elkyl substituent end men s substituted bensyl group further changes in the elkyl substituent would be of interest. The purpose or this stucb' is to propere s series of Emma» saline hydrochlorides containing s cycloelkyl group and en srslkyl substituent. In this work the oyebelkyl group is the mlohenql end eyolohmlnetlwl group while the erellql substituents ere the bensyl, substituted bensyl, big-phavlethyl end m-phenylpropyl groups. The structure of the compounds is besed on the smiley of the methods of synthesis employed with those described for the properstion of Wotlcnomuructuresndisflmtherestsblishedby eomperieon of their inns-red ebsorption spectre with spectre of mem- structures reported by Percivel (27). 35 DISCUSSION The preparation of l, hodisubstituted ScimOtetrasolines my be seconaplished by slkyletion of locubstituted S—aminotetrasoles with either alkyl halides or alkyl bensenesulfonatee (1,2,28,29). In earlier work (28,29) the product of this reaction was thought to be l-alkyl-Salkylandnotetraaole. Recent investigations (1,2) have shown that the product resulting from the alkylation of e. l-elkyl-S-amino- tetrasole is a l,h-dialkyl¢5cininotetrasoline. Percival and Herbst (l) have elucidated the structure of the alkyletion products or l-alkyl-S—aninotetrazoles in the following manner. By introduction of the group R by alkylation of leaks-sum. tetrasole a product ins obtained which was identical with the product obtained men the group R' was introduced by alkylation of l-vRoS- aninotetrasole. ‘ ENE—MB. HRH—Olen 0H1 ' RAY—rim. 3‘”)! \N/ \/ HrPd (when R «- bensyl) man—fem, N\N /N 36 rheseresnltseouldberealisedenlyiftheproductofalkylatien contained substituents situated in equivalent positions, that is , the l and 1. position of the tetrasole nucleus. Additional evidence as offered by the catalytic hydrogenolysis to remve the m1 group from a Me:- of l-flhl-h-bensy‘l-E-Mnctetraeolines prepared either by alkylation of l—benzylnSoaninotetrasole or by bensylation of s l—flJql-S-Mnotetraaole. In every instance lvalkylo5~aninotetrasoles were obtained from the debonvlation reaction, a result which could ariseonlyifthe ametituentsarelooatedintheoneendtourposi- tions or the tetrascle ring. Thus the reaction or an alkyl halide with a l—alkyl-S-aninotetrasole my be represented by the following equation: NH, NH on net/Kn e an: ----> R-N/KH-EU fl—H an The 5-innotetraeoline Wdroohlorides described in this work were prepared by alkylation of l-cyclohecql- and l-cyclohexylmthylos-elino- tetrasole by the method described by Herbst (30). The W introduced by alkylation were the bonnyl, p-chlorobenzyl,o-chlorobemyl,2 ,h-di- chlorobensyl, 3 ,h-dichlorobensyl, p—nitrobensyl, n—nitrobensyl, RES" pherwletlvl and m—phenylprcpyl. Of the group 01‘ ininotetrasoline morochlorides prepared only l-cyclohwluhébemyl-S-ininotetrasolino hydrochloride had been described before (29). in the some paper is a 37 report of the preparation of l-cycloheaquuh-bgtg-phenylettvl-S-Mo- tetraaoline hydrobromide. In the preparation of the imnotetraaoline hydrochlorides the l—alkyl-S—amjnotetraeole was mad with the appmpriate m1 halide and heated on an oil bath he lBO-thOG. for three to eight hours. The use of about a 50% noose of the alkyl halide facilitated the formation of a homogeneous nelt during the early part of the heating period and probably increased the yields. After the heating period excess alkyl halide was removed by steam distillation and the residual product separated fron any unreected l—eJchl-S~aninotetrazole by conversion of the innatetraaoline hydrochloride to the free base and removal of the free base by extraction with other. Finally the insinc- tetrasolins was again converted into the hydrochloride and further purified by recrystallisation. During the extraction of the free bees with other it was found that good separation of the indnotetraaoline was somewhat difficult to mowlish. Apparently the inimtetrasoline hydrochlorides , which are mt very soluble in water, do not react very rapidly with the aqueous base to liberate the free ixdnotetrasoline. Besides this the free ininotetrasoline forms a very viscous oil that costs the moanted hydrochloride thus mung liberation of free base even more difficult. In order to extract the indnotetrasoline with other the mixture met be shaken for a long period of time. Addition of a few milliliters of alcohol to the contraction mixture will make separation more rapid but this appears to increase the water content of the ether extracts. 38 The free bases whim were liberated from the hydrochlorides were pale yellow, viscous oils in nest instances. In the case of five inino- tetrasolines which had eyolchexylssthyl substituents the oils could be crystallised to give low melting, colorless solids . The free bases forned Wehloridu in either aqueous or alcoholic solutions and were readily condensed with phmyl isothiocyanato to give phenylthioureas . This behavior is in direct contrast with the properties of l—alkyl-S- alkylauinotetrasoles (l) which do not condense with phewl isothiooyanate. the l-eyclohuyl- and l-eyclohssylaetlleS—auinotetrasole used in this work were prepared by a method similar to that used by Garbrecht and Herbst (31) and an be represented by the following scheme: ‘lms Brow m4 / R-HH. W R-NH-CN ”—8.... 3-K 7 1. E” All of the steps were carried out successively in the same flask without isolation of any of the intermediate products. In carrying out the reaction, amine hydrobronide is formed in the first step along with the eyansside but addition of an equinolar amount of sodiun hydroxide allows all of the anine to be converted into the desired cyansnide. The final step in the reaction as sham in scheme 1 was carried out by addition of a sodiun aside solution to the reaction mixture followed by hydrochloric acid in order to dispense with the necessity of preparing and handling separately solutions of hydrasoie acid. Presumably the reaction of hydrasoic acid with a cyananide leads to an intermediate guanyl aside 39 which cyclises upon heating. This intermediate could conceivably cyclise in either of two aye to give a l-alkylc-S-aninotetrasole or a Scalkylaldnotetrasole, however, in all instances reported the only product obtained as a l-alkyl-S-aninotetrasolei even whn the substituent poops, B, were of quite different electronegativity, nah as methyl and 293- nitrorhsvl (31) . l—Cychhml-S-mmtetrasole had been described before (29) but in order to further characterise the hitherto unkneu loeyclohesylnetrwl-Se- admtetraaole a mall uncut was converted into l-cyclohuzylaethyl~5- acetylaninotetrasole by heating with acetic anhydride . The cycloheaylnethylauine used to synthesise l-n-cyclohesylnethyl-S- mnctetrasole was prepared by none of a Schmidt reaction fro: cyclo- hecylaeetic acid. A solution of one uncle of earbcaquie acid in nine soles of sulfuric acid us employed and the hydrosoic acid was added in the fora of a 171 bsnssna solution. although the scale of tin reaction assaswhatlargerthanisasaallyenoounteredUZ) noseriousdiffia cultiee were set with. The reaction as strongly exothermic but the temperature could easily be one below 50°C. by control of the rate of addition of the hydrasoic acid or by use of a cooling bath. In more recent applications of the Schmidt reaction on a small scale (32) solid sodiun aside no added to the reaction mixture but, in the present case, the use of a bensene solution of hydrasoio acid made possible easier control of the reaction temperature. Isolation of the amine was facilitated by the formation of an insoluble amine salt, probably the acid sulfate, upon dilution of the sulfuric acid with water. Removal of to this insoluble tutorial made the neutralisation of the large amount of sulmio acid unnecessary in order to isolate the free sadne. This procedure appears to offer a good nethcd for the preparation of aliphatic amines in those instances where the corresponding carbosylio acid is available. The infra-red absorption spectra of a wiser of the innotetraeolinss and their hydrochlorides were obtained in order to distinguish these compounds from other alkylation products which could arise. mutilation of these spectra revealed several notable features. The mnotetrasol inc hydrochlorides shoe very strong absorption at about 5 .95 p and a notable lack of absorption at 2.9 - 3.2 u and at 3.7 e 14.1; m the regions usually “scouted with ll-H vibrations and amine hydrochlorides , respectively. In the spectra of the free bases the dominant features are the absorption band at 3.0 p and the strong band at 6.5 ll. A study of the infra-red spectra of a series of tetrasoles consist- ing of S-alkylanimtetrasoles, S-dialkylaninotetrasolss, l-alkyl-S-anino- tetrasoles , l-alkbe-alkylaninotetrasolss, laslkyl~5eslkyluinotetrasole hydrochloridee and l,h-dialkyl-5-iuinotetrasoline lvdrochlorides by Percival (27) revealed several absorption bands which can serve to disc tinguish one from the others. For instance, l-alkylas-allcylmiwtetrasole hydrochlorides can easily be distinguished frou l,h-dialkyloS-isdnctetra- saline hydrochlorides by the strong and rather broad absorption band at h.o .. h.h p which appears in the spectra of l-dhl-S-alkylasinotetrascle hydrochlorides. The lacyeloalkyloh-aralhl-S-ininotetrasoline hydrochlorides pre- pared in this work show spectra very similar to those of the l,h-dialkyl- momma hydrochlorides reported by Percival (27). Their lack of absorption at 3.7 - 11.14 )1 indicates that they are not l-cycloalkyl-S- aralkylaainotetraaole Ivdrochlorides, even though there are new sinie larities to the spectra of l-alkyl-S-slkylasinotetrasole hydrochlorides reproduced by Percival. Pros an inspection of possible structures of themdrochlcrides euchasthoseehominlandnonesightexpectsone similarity in the infraored absorption spectra. , a 1’ R-l'l/ \f-H 01" n-s/ \s-e' 01" 13—3 u.“ I II Both of these compounds would exist as resonance twin-ids as a renlt of omnidiniun ion type resonance as shown in III, IV, V and VI. Hy $1! 11“}! a“ )1 Fl“ )1 g 0 /¢|3 A e 34/ NM: 4-. NV NH: 4-—e- a-u/ \ .3 4—. MM \r-n .=B H=N N=£l :l=' III Iv v n The infra-red spectra of the free 1, h—disubstituted-Scimnotetra- salines have several characteristic features which serve to distinguish 142 then Iron the l-flkyl-S-fllqhflmtetrusolee. The most prominent of these features is a strong band at 6.03 p in the ininotetrasolines mm. the l-eJJql-S-anqlmdnotetruolce exhibit no absorption at 6.03 p but have a strong band at 6.28 )1. Some work on the infra-red absorption spectra of a umber of tetra- soles hae been reported (27,33,314) and attempts made to aeeooiete certain structures with epccific absorption benda. When one We these aeeignnents some uncertainty becomes apparent. In the studies reported by Lieber at a}. (33) absorption in the six micron region show by Sraninoo tetraeole wee attributed to the amino group but in the same paper absorp- tion at 5.95 to 6.02 p present in the spectra of guenidines was attributed to the imino group. Hurptw and Picard (31;) have examined the spectra of l,hodimt}vl-Sdmnotetrazole and have assigned the band at 6.0 n to the We pomp. A suggestion has been made by Percival that the absorp- tionband at 6.0nnaybe due to etmctures suchasVII uhiletheband often found in tetruaolee at 6.3 u could arise fro: structures like VIII. u 1: ‘IL I n/ a n’c‘n l l I I 3—3 13—3 VII VIII In some aninotetraaolas often uritten as VIII, resonance ferns involving structures such as VII may be written with charge separation and thus impart inino character'to the molecule. The bends at 6.0 n and 6.3 )1 may then give a unsure of the relative contribution of each resonance form. h3 Hem-y, Finnegan and Lieber (2) have alkylated a series of l-alkyl- S-aMnotetrasoles and reported that the major products were l,hodiu]kyl~ 9mm hydrochlorides, however , they reported the isolation of a small mount of a second product which they describe as a nesoionic compound, 1,3-dia1kyl-5-ininotetrasole II. RH NH II II /C\ /c\ ‘“' ‘”’? “f ’i X 3-3 Ran—L II I This type of eoqound has bean reported to result from the allqlntion of 2-a1kyl-5-aninctetraaolee (3) e The structure assigned to the compound as based on data obtained from bray diffraction studiee and its chemical properties 3 however, the chemical evidence offered could also be explained on the basis of a l,2~dia1kyl-S~ioinotetraaoline X. In the alkylations conducted in this work no bywproduct was found in either the tetraaolinee or their condensation products with phawl isothiocyanate. EXPERDTEII‘ITAL Prgparation of mlohfllmetrglergge Cyclohexylnethylandne was prepared from cyclohemylacetic acid by loans of s Schmidt reaction. The procedure was devised from the data reported in studies by Oesterlein (35) and by Schuerch and Huntress (36). A 5 l. flask was fitted with a reflux condenser, alcohol thermometer, stirrer and 500 nl. dropping funnel and set up over a cold aster bath which could be elevated in order to control the temperature of the con- tents of the flask. The flack was charged with 21.3 g. (1.5 moles) of cyclohexylacetic acid (b.p. 139-1ho°c ./‘17 m.), 2500 mi. of benaene and 715 :1. (13.5 noles) of concentrated sulfuric acid. From the dropping funnel, MS ml. of a benzene solution of hydraaoic acid containing 17.0 g. of hydrasoic acid per 100 ml. of solution (1.8 moles of hydraeoic acid) was added to the reaction mixture with vigorous stirring at a rate of about 3 ml. per limits. in exotherdc reaction took place but the temperature of the reaction mixture wue kept between h2° and n8°c. by adjustment of the inter bath. After complete addition of the hydraaoic acid solution the reaction mixture was held between Mo and h8°0. for one hour by gentle turning on a steam bath. The contents of the flask sore cooled in an ice bath and the eulfuric acid Ivor separated iron the bonsone Lever. DrOpping the sulfuric acid solution into a h l. beaker filled with crushed ice resulted in the formation :1 a dense white precipitate. The precipitated audne salt was 145 filtered rapidly with suction, pressed as dry as possible and immediately transferred to a h l. beaker. After airing the solid with l l. of later the mixture was made alkaline by the addition of about 600 g. of potassium hydroxide in the form of a 50% solution. Il‘his treatment caused the separation of a pale yellos anus layer which was separated from the aqueous residues by steam distillation. The amino nae separated from the water of the distillate by extraction with ether and the etherial solno tion dried over potassium carbonate. From the ether extracts there was obtained, by distillation, 131 g. of cyclohencylmethylamine, b.p. 162- 163°c. at atmospheric pressure, up” l.h632 (37). The yield based on cyclohmlacetic acid as 68%. A small portion of the sulfuric acid filtrate, from which the mine salt separated, as ands alkaline by the addition of 50% Potassium lvdroxide. Steam distillation followed by extraction of the distillate sith ether failed to reveal any appreciable amunt of amine. l-flkyl-Scindnotetrasoles Preparation of l-cyclohu'yl- and l-cyclohelqylnethyl-S-aninotetra- soles was acccnplished by a method siMlar to that reported by Garbrecht and Herbst (31) . ' 1 10h. 1m (1" tetrasole A solution of 113 g. (1 mole) of cyclohelqlnethvlanine in 800 ml. of ethanol was cooled to about u°c. in an ice bath. A solution of 1.06 g. (I. able) of cyanogen bromide dissolved in 1:00 ml. or 501 ethanol see added dropsise, with stirring, at such a rate that the temperatmre of 116 the reaction moire remained between 8° and 10°C. with the temperature of the reaction mixture still maintained at 8° to 10°C., to g. (l sole) of sodiun muronde dissolved in 200 ml. of water was slowly added. The reaction mixture see then treated with 81 g. (125 moles) of sodium aside dissolved in 250 ml. of water followed by dropuse addition of 210 ml. of 6 l mdrochloric acid (1.25 moles). Addition of the hydrochloric acid was adjusted so that the temperature of the reaction did not rise above 12°C. After addition of the deroehlorie acid solution the reaction adxture Ins refluxed gently for three hours. At the beginning of the reflux period a white precipitate began to fern and after half an hour of heating precipitation appeared to stop. The flask was cooled after reflnnng and the product removed by filtration. This gave 1&5 g. of fluffy white needles of lc-cyclohexylnetlwl-S-udnotetrasole (80; yield based on cyclohelqlnetlvlanine) melting at 250-25100 . Analysis: Calculated for cerium: c, 53.0%; H, 8.3%; N, 38.6%. Found! C. 52.9%; H, 8.3%: H, 38.8%. in acetyl derivative was prepared by gently remixing l g. of l—cyclohsxylmetrwl-Soaninotetrasole with 2 ml. of acetic anhydride for about 15 urinates. The remlting l-cyclchmqlnetlvl-‘S—acetylalmotetra- sole was recrystallised from 50% ethanol giving a product melting at lie-130°C. Analysis: Calculated for 0,931,1150: G, 53.8%; H, 7.751 N, 31.1%. round: 0, 53.7%: H, 7.6%; N, 31M. s Analyses were done by Micrco'l‘ech. Laboratories, Skokie, Illinois. h? Preparation of l-cyclohexy1-5-aninotetraacle in the some manner as that used for the preparation or lpcyclohmlnethyl~50aminotetrasole gave a 62% yield of a product melting at 217-21806. (29). l,h-Disnbstituted S-Mmtetrmline Hydrochlorides These someondswereprepared accordingtothe method givenby Ila-bet (30). The appropriate l-alkyl-5veainotetrasole as mixed with an elkylhalide andheated onanoilbsth at Jae-116°C. Masher!» period of heating a viscous solution as termed mien slonly solidified. Heating was then centimed for tron two to six hours after the melt had coupletely solidified. The erade asterial us dissolved in a hot alcohol- uter nature and subjected to steal: distillation in order to relieve: lamented alkyl halide. The residual product was made alkaline and the free inductetraseline attracted from the basic solution with other . Evaporation of the ether extracts left the crude innatetrasoline which as dissolved in aqueous alcohol and converted to the hydrochloride by the addition of hydrochloric acid. This product was then recrystallised from aqueoas ethmol. Alkylating agents used were substituted bensyl chlorides, Wopheiwletrwl bronde and m—pheiwlpropyl brolide. Maples of the method of preparation are given in the following preparations of locycloheuwl-h-p-chlorobensyl-S-Mnotetrasoline hydro- chloride and lucyelohaxylmstlvl-hwdphexwlettvl-S-inimtetrasoline wdrochloride. M3 1 cloh 1- hlorob 1- iminotetr line chloride 1 sixture of 8.1: g. (0.050 mole) or 1-cyc1oheoqyl-S-aninotetrasole and 12.1 g. (0.075 uncle) of p-chlorobenayl chloride was heated on an oil bath at 11.0%. After heating tor about half an hour a homegeneous melt formed which slowly solidified over a period or about half an hour. Heating was continued for two hours after the melt had become oozzpletely solid. The solid tutorial was remved by dissolving in about 200 ml. of boiling 501 aqueous ethanol. The alcoholic solution was diluted lath eater and subjected to stean distillation. After the distillate cane over clear, the residue was made alkaline by addition of h.0 g. {0.10 nole) of sodium 1vdroxide. The alkaline solution as shaken vigorously for approximately half an hour then extracted with ether. In order to remove all of the free iminotetrasolinc three portions of other were used and in each case the mixture one shaken for about 20 minutes. Evaporation of the ether attracts left a yellow oil which was taken u) in 50 all. of ethanol. A pale yellow precipitate was produced upon acidification of the solution with concentrated hydrochloric acid . in additional 50 al. of ethanol and 100 ml. of Inter was added “to the mixture and the precipitate dissolved by heating. Digestion nith Horite, filtration am subsement cooling produced a crop of colorless needle- 1ike metals. Recrystallisation of this product iron 50;! aqueous iso- propyl alcohol gave 11.6 g. (70% yield based on the aminotetrasolc) of pure colorless crystals of l-cychhcmyl-hvp-chlomhenayl-S-Mnotetrao saline hydrochloride melting at 229-23o°c. with accompanying decompo- sition. 1L9 1-Gycloheoqylnethyl-h-beta~pheglettwl~531minotetrasoline hydrochloride A mixture of 9.1 g. (0.050 mole) of l-cyolohelqlnethyl-Soeminoc tetrasols and 13.9 g. (0.075 mole) of iota-phenylwhyl bromide no heated in an oil bath at 115°C . The mixture gradually two a homo- geneous melt that slowly solidified over a period of about one hour. Beating was continued for six hours after the melt had completely solidified. The product use dissolved in hot 50% ethanol then diluted uith enter and subjected to steam distillation. than the distillate began to cone over clear the residue was nude basic by addition of h.0 g. (0.10 mole) of sodium hydroxide. The alkaline solution us shaken vigorously for half an hour then contracted with three portions of other uhich were also shaken for about half an hour. Upon evaporation or the ether extracts a brownish oil rmined ehich res dissolved in 50 ml. of ethanol and converted into the Wdrochloride by addition or com entrated twdrochloric acid along sith 5'0 ml. of water. The on de hydrochloride was dissolved by heating with an additional 100 ml. of water and then digested with florite. After filtration and cooling a fine colorless precipitate formed . A ascend recrystallisation tron 20% aqueous isOpropyl alcohol an 10.8 g. (67% based on the aminotetrasole) of laoyolohexyb ntlvl-hfigtyphenylethyl-S—ininotetraaoline hydrochloride melting at est-235°C . with decomposition. All or the ininotetraeoline hydrochlorides were prepared in the some runner and are listed in Tables I and II along with descriptive data. Analytical data are listed in Tables III and IV. 50 TABLE I imam-h-mn-sammmlzm HYDROCIEORIDE RH 'HCl 0\ m a 3.9303 title KW no: i A; A .5 __ i "m“? All in. A.“ Benny]. 230 72 50% leopropyl alcohol p-Chlerobensyl. 229-230 70 50% isoprcpyl alcohol e-Chlorobusyl 222-223 5h 50% “ml alcohol z,h-niohlorooanayl 239-236 lo 60% impel alcohol Lit—mm 219-220 58 60% W1 alcohol p-litrebenayl 21.1-21.2 Sh 70% isopropyl alcohol n-Iitrobenayl 217-218 61; 70% isopropyl alcohol maul ,_ 220-221 53 . 25% tum-owl alcohol m-Phenylprcpyl 222-223 Sh 25$ isopropyl alcohol ___.__A-__._a__4_ _l__ni_i___ A M.“ 4 _.._._ Ail—4 m fiv—wv i F7 ‘ w ,. W vw— v—w —'—v ——--—— W wV—v—w All compounds melted with accompanying decomposition. TABLE II leCIGLOHEXILMETHIL-h~ARALKIL~5¥IHINOTETRAZOLINE HYDROCHLORIDES NH 'HCI 3\ (cyclo)cen,,CH,-T”’ W fi ..... w. ..v—V W W Benayl 217-218 61 SOZ’isopropyl alcohol peChlorobenayl 210-211 70 50% isopropyl alcohol o~Chlorobensyl 23hr235 58 50% isopropyi alcohol 2,h~Dichlor6bonsyl 220 66 60% ieopropyl alcohol 3.heDichlorobcnsyl 216 73 70% isopropyl alcohol p-flitrobennrl 232 71 80% isopropyl elcohol uwflitrobensyl 213.219 65 60% isopropyl alcohol [gaggerhenylethyl 23h~235 6? 20% isopropyl alcohol w—Phewlpropyl ZhOthl -. 69 201 isopc'opyl alcohol , , MW? n_ii_i‘_ i:i4i #w lfi;_ : ipri All compounds melted with accompanying decomposition. 52 .35 £33» {toe-no.3 €8.23: B 88 at: 223.5 . . a t a. mod EN «2: ad o.» m4. 2% mg“ azaleas “Edging 0.3 to no on S. 3. new on iconic ages; are «in we” ma." .3 Rm We. 49 aoofiuamiu gen... 3“ 04a Q3 m.2 em 9m 33 to: aoansamzo aha-8.32..“ «.3 2a on to... o.m a.“ «.3 3.. £535 1.8.3833; 33 33 4.3 3n dm 9m 4.3 4.3 onosanzo can-33.4.“ «.3 «25 dd 0.”... 3 9m «:R «Am {Gauze analog: 4:3 «.8 ed ed ad, ad ”Am «Am {doodle age 0.3 fins «.fi 33 a.» a6 hem «am anBoamae 328 do... aggooefim ggga .6 flag on a 53 .38an {none 835333 «8.789.: .3 8% at: 3.3.5 ‘ .l 1.8 «.8 «.3 «.3 «4. «.« «.8 «.8 ozGoamao «Ragga-a ode «AN «.d. o.§.. m.« m.« mdm «.«m oasoamooo Hgfifionatmmmm «.9 «.«u 0.0.” 0.3 .2 o.« «Am «Am acofionamodo Agatha... «.3 «.9 «.3 odd hm o.« «2G 4% aoofioaamflo «gonad «.«H 0.3.. «.mm «.«m :.m rim 43 «.3 oznfloemouo ”goggtaan «.«H «.«a m.«« ~.«~ «.m dm «.2 «.5 ozosoaefio Hangoaofloaéam «.8 «.2. 0:8. «.8 «.e «6 «.«m «.«m guacamole «gaze... «.3 «.8 «.8 «.8 «we «6 «.«m «.«m ozossmflo «Ethanol «.mu «.«m «d «:2 «4. «.« «.«m m.«m ofiooamso «boom one 8 «EB! 13mm We? I 1&2 m“ :H Ewan «than . I t r 1 11 1! z pooch." 8 «595% «cotton «photo «Baton .o. _ _-. s . «.2/ \2 mo: «cannons Sn. «2 omegéoommé «fidgggltfiafitffimfigogol no finned: «H «3:. 524 Derivatives With Phenyl Isothiocyanate All of the iminotetraeolines were characterised as phavlthioureas by treatment of the free base with phemrl isothiocyanate. The prepara- tion or the phemrlthiourea derived from l—cyclohexyl-h-p-ehlorobcnayl~ 5-iminotetracoline is typical of the method used. Ph ltfihiwourga derived#rron#_l-§Elohfll-ymrob cal-Evindnotem- IO About 1 g. of l-cyolohmcyl-h-p-ohlorobensyl-S-ininotetrasoline murochloride was shaken vigorously with 5 ml. or 2 1! sodium hydronde solution. The resulting iminotetrasoline ms extracted with other and the ether extracts dried over sodium sulfate. After removal of the dry- ing agent the other was evaporated on a were later bath. A viscous oil rwained which see treated with approximately 0.5 g. of phenyl isothio- cyanate and heated on a steam bath for from 5 to 10 minutes. The yellow oil so obtained sac crystallised by stirring with 5 ml. of hexane. Recrystallization from isopropyl alcohol gave a colorless product melting at 153.5 to info. All phenvlthioureas were prepared in the same mnner and could be recrystallised from either icopropyl alcohol or heptano. Melting points and analytical results are given in Tables V and VI. l-Cycloharylnetrwl-linaralkyl-S—indnotetrazolines The tree bases were prepared by neutralization of their hydro- chlorides. The procedure used may be illustrated by the preparation of l-cyclohexylmethyl-h—pwhlorobenzyl~5~iminotetrazoline. About 1 g. of 55 .853 633m .BCSEBS hSBéoE B 83 ch... 8.33 i Hun it 1AM 4,11 .4111 111 3. 3. ed... cam 1. a. mozflmao tact? Heefiaeuafi..§ a; m.» ”.8 wow I. I. «maniac Seneca afipoabflfiuflon «4. 3. 5w due a... 1. Qoiamao mags” ”beneath: 3. 3. SN flow 1. . I encased; m.§um.8a abfinobezh me 0; mad «.3 4.3 4.3. nee-83m“; sedan aghafigfiaéa 3. 0;. m4: «.3 a «.3 4.3 amalgam"; Nfiéfi Hedonfiofluauim 3. m; 53 we” ed ma woman-mac maneuméen eggs: .3. m4. mg? 53 H5 m5 mofioamse 33:35 $5851 06 «a «Aw dd . a... I. amazed-e 93:: ”been 0 i 1 a 1 i .1 1 1 _m age. 8K a m 2 ea: 83 8 remote Japan .003: a ”nil .. .3 e m z/o\z.. m 035v encode-8..» dqggggilagogd 5E gm gogugm bag 56 .35 dream .3fl3333 essences E 88 22. Bang t. 3. .3. mé n.3, t. 2. memorize 375 HEB—8% m.» . e.» «.8 0.8 I. I. mozamfle $3.8 nonfiéfitmfim o.» 3. ed pea .... .... niacin-mane 6.1.3 ”Peg—gang 0;. a.» 9.8 ha I a... enormfimflo $33 Haggai We we 95 SH 0.3 .34. men-Gtmane 8.32 ”58358.44 me we 0.5 3.." m3: m4: mafidxmae “unaided agfioafiaid oh. 2 add 43 06 ed manganese «3.15 ”95063395 «4. n.» odd #3 «4. 9m eeaamao m.em7m.mma Hawaii m; .3. o4.“ v.8 i. I. momoamn-o méfinflnfi $38 353 .0... a: u a a ML/ \fiamVSmeofioHfis o engageoémégfiégoga 20E Emma gage H» mama . 57 h.@« .35 :35 .SgnBa-A 6.9.83. h. 83 98.. .3334 L You I. 1. e6 .3 QR «.3 nooufimSo $18 v.8 v.8 fies 98 ».m 9m 9% 9mm Juana-nu 2A.» abfiaouqaéfi wow v.8 ad 98 «am o.m 3% 0.8 {Gan-«a 3.78” ”5.8354.“ dam m5.“ .... I. 5,. ed 3m 3m wanna—ac no.3 “Ego...“ Wm“ can I 1. a.» a.» 4.8 4.8 nun-man Wanna Hg inch 338 rim.“ mung mg «1.36 Him 2381 . 1 r . a tn........_ .. _ ....B «:83. m. I Egk m $898; 0 338m B ”an l-cycloheorylmethyl-hvp-chlorob eml-S-iminotetraaoline hydrochloride Ins shaken vigorously with 5 ml. of 2 N sodium hydroxide solution then extracted with other. The ether extracts were dried over sodium sulfate and then the other evaporated by means of swarm water bath. A viscous pale yellow oil resulted which solidified upon cooling in an ice bath. Dissolving the crude solid in hot cyclohexane followed by very slow cooling gave oolorleu needles of l-cyclohexylmethyl-hdp-chlorobensylu S-iminotetrasoline melting at 82-83%. Attanpts were made to prepare the free base. from all of the hydro- chlorides, however, only five of the compounds gave solid products. The solid compounds isolated are listed in Table VII along with their melt- ing point- and analytical results. Infra-red Absorption Spectra Infra-red absorption spectra. were obtained for a series of imino- tetraaoline hydrochloride: and the corresponding free bases using a. Perkin-Elmer Doublebeem Recording Spectrophotometer, Model 21. All compounds were run in oil mile with the concentration or solid great enough to five strong adsorption in the six micron region. The spectra are reproduced in Figures 2 to 16 of the Appendix. 1. 2. 3. S9 SUM-SARI The reaction of 3 ,S-dimethyl—h-sminoc-l,2,h-triazole with bensyl chloride was shown to lead to a quaternary chloride which contains a primary shrine mup. The maternary chloride has been shown to react with aqueous potassium hydroxide to give an organic base that forms a Ivdrochloride isomeric with the quaternary chloride. Treatment of the base with bmsyl chloride results in the formation or a second quaternary chloride . A series or l—cyolcalkyhhoaralkyl-S-imotetrazoline hydrochloridee has been prepared by the aralkylation of l-cycloalkbe—Wtetrazolee. The compounds were characterized by formation of phewlthioureas by reaction with phenyl isothiocyanate. The structure of these compounds was established by comparison of their infra-red spectra with the spectra of know. structures and by analog of the method of synthesis with that described for the preparation of comounds of known stmcture. LITERATURE CITED (1) a. a. Herbst and D. r. Percival, J. Org. Chess" 19, 1.39 (1951.). (2) a. 1. H w. 0. mm and n. Liebsr J. in. Chen. see. 6 2891. (1:31:33 ' ' 1" (3) J. Dryden, R. A. scary, w. 0. mm, a. Beeches, w. s. 11ch and a. 2. Van Dolah, J. n. on... 800., 15, 1.863 (1953). (u) 3. Stem, J. prskt. one... [2], pg, 1.68 (1903), Chas. antr., 7h. 793 II (1903). (5) a. sun‘ and x. rho-a, J. prskt. one... 1;, 288 (1906), Chen. Zentr., 1906, 1783 I. (6) ggostons, J. prskt. one... 35, 1.16 (1907), Chen. antr. 1201, O (7) s. Ruhmnn and H. Staplston, J. Chen. 8cm, (3;, 261 (1902). .(8) 8. Kuhn-um, J. Chem. 300., 92, 1268 (1906). (9) R. H. Herbst and J. L. Garrison, J. Org. Chem, 1g, 872 (1953). (10) H. Clarke, H. Gillespie and s. weisshaus, J. is. Chem. Sec" 55. 1:571 (1933). (n) s. Wagner, J. in. Chen. Soc" 55. 721. (1933). (12) L. J. Bellamy, 'The Infra-red Spectra of Complex Molecules,“ 1143;?» & Co. Ltd" London; John Wiley & Sons, Inc. , New Iork, ('13) x. s. Colthup, J. Optical am. on... 59. 397 (1950). (11.) L. Birkofer, Ber... 12, 1.29 (191.2). (15) w. 1.. Garbrecht and a. s. Berbst, J. Org. Chem... 19, 1022 (1953). (16) a. n. Herbst and w. L. Garbrecht, J. Org. Chem, 19, 1283 (1953). (17) 1. Mom, 3.2., 31, 98h (1891.). (18) ‘s m, Ms, M, 221 (1897)s (19) o. Silberrad, J. Chem. Soc., 11, 1185 (1900). (20) r. Curtius, L. Darapsky and 3. Inner, Ber... (Lg, 1191. (1907). (21) 3. Phillips and A. Hichaelis, 11711., 322, 292 (1889). (22) 1. Honor, Ber., 29, 1871 (1897). (23) 8. Ruhemann and R. Merriam, J. Chen. 80s., 21, 1768 (1905). (21.) 0. Rules and J. Weber, Ber., 1.2, 2715 (1909). (25) H. H. Hatt, "Organic Synthesid' Col. Vol. II, John Wiley 8: Sons, 1110., Hear York, 191.3, 1:. 208. (26) '1'. P. Reutner, J. C. Peters and E. P. manger, “Abstracts of Papers Presented at the 129th national American Chemical Society Hesting,‘ Dallas, April, 1956, p. 714. (27) D. P. Percival, IIA‘.l.11:3,'1|.a1'.ed S-Aninotetrasolss, “their Preparation and Properties," Ph. D. Thesis, Michigan out. college, 1955. (28) J. 21.1.1. and a. Ingle, Am... 291, 233 (1895). (29) B. H. Herbst, C. U. Roberts, E. J. W, J. Org. Chem, 16, 139 (1951). (30) a. H. 301%“, u. 8. P‘M 2,735,852. (31) U. L. Osrbreoht and R. H. Herbst, J. Org. Chem, 18, 10111 (1953). (32) R. “011', 'Organio Reactions,” John Vile: 8: Sons, New York, 19119, PP- 307’3350 (33) %. 1333b», D. Levering and L. Patterson, Anal. Chem, .2}, 1591. 19 s (31.) D. 3. Murphy and J. P. Picard, J. Org. one... .12, 1807 (1951.). (35) u. Oesterlin, 2. mm. Chemie, 352 536 (1932). (36) c. Bernice-ch and s. Huntrsss, J. Am. Chem. soc., fl, 2233 (191.9). (37) J. (Int, Ber., g9, 2065 (1907). ' APPENDIX 62 otCoEchGmm o:::Nm.S$oEETm-3min-endkxcchkoé T 1:173;ch 6.1.7.955 .m 9:3; Amconuficv 2323333 2 2 . w w e we _ l _ . l n _ A _ d A _ o 2 .1 ON 3 IL on I ow _ _ _ _ _ _ _ H V _ _ cg uogssgwsuedl wag) .Iad 63 E mUEoEooSmm mcficambfloEETm-3552803047wiaxwzofiomo-H mo 85.5095 vogébcw Amcouoficv Emcflgmg NH 3 . m o .m mpsma m.N q. _ _ a _ _ _ a _ ON 9‘ 8 cm 2: uogssnnsueu, mag Jed 64 3 mUEoEoounwm mczoumbfioEE79235280226-¢ .mv-v-3xm:o~o>0-~ Ho €5.30me 6972.2; .v 81.th NH 0H Amceowfiv fiwcggag w md _ _ _ -Mo om ow ow cm 02 uogssgwsuml wag .Iad 65 QEEELOLFE 2:.Emgcgsficfi-mAamconochczYv .mv -wékwnficfinué .3 2:559on to.Ta.£E. .m 9.53% $203.15 53:29me 3 NH 3 w m .m fl _ A _ _ ~ A ll '1 om ow ow cm 2: uogssgwsuml mag .Iad 66 E ociszoogpmm oczcmmiofiEEzum-3§5222:17wafmxozcflo‘mnVuH we Esbuogm Ucp1mtg «H OH $20525 Ego—gap» .w 93th rim L om ow om .ow .2: uoxssywsuml mag) 13d 67 ,VE.:::.5,€»: BEQNN.7.Q:::.ETm:3.57.2-7TEZEmegscH3.0-H .5 Esfigrim UPthE 3:97:25 Eucofiémfi .~ IL chain E 2 2 w m m .N fl a _ 4 _ A ~ _ _ _ 3 l . l r _ _ b _ b _ T om o¢ ow cm on: uogssgmsuell wag) .Ind' 68 E mEchuobfim oEEambBoEETm-ENcono.~oEo-;-wémfaEExmnofiomU-fl Ho Esbomam @97th NH 3 6:082»: 5329me m .m «£53m _ _ _ on ow. om ow OS UOISSIILISUBJJ. 1113.4 .183 mnCoEoougm mczonmbmgofififm-AgucmnouoEfiné .8-wtfifimEExmongU-H we 52.20QO vermbfi .m magma mu, 320.825 533953 3 NH 3 m o a. m.m _ _ A _ _ _ _ d d _ _ o 3 TI .1 cm [I 3 1 av II I. on 2 I. C .I ow _ _ _ _ H _ ._ _ P _ _ 2: uogssuusueu, mag) Jad ochEELEE 250ngonngTm-:>.N:¢Qc.HcEg€uv .mvlv,fifoEixocofaunH E Estoogm @9722: .3 Snug W 3:83;: 5m53>m3 3 NH 3 m w H. m .N _ _ _ _ _ _ _ _ _ _ _ o _ _ _ _ _ _ _ _ _ _ _ o2 uogssgmsueu, wag) .Iad 71 mchEoouEHm 0233285587muauconobE-a-vuafimEaxmsoHumO-H mo 85.5095 3.7985 .HH 93mg AmcouBEv Swag/«3 «H NH 3 m o v m .N _ H d H _ _ _ _ d _ 3 _ T'- l r... 3 III ON 3 on cm 2: uogssnusmaq, wag 18d 72 953235587@173onv-§fm§3§noe§0-H Ho 9253QO 60.7955 .NH ousmflh .Amcouuficv 5320.253 «H NH 3 m w m .N _ _ _ A _ _ I J I 4 _ _ _ _ _ _ ON 3. oo om ooH uogssxmsueu, 11133 Jed 73 oczoumbgocHEH-mANmEQPSEuATTHEEEExmonozU-H O0 Eahuahm 60.799: .mH mama $20325 EmangmB vH NH OH O O HV _ O _ H _ O H . _ 1| 1 s _ _ H _ _ _ _ _ ON OH. OO Om OOH uoIssuusueJJ, mar) .Iad mczcmmmeoEETm-AENFEQSEBUQ .Nvéu35087382230-a Ho Esbumam 8.7.925 . $5.825 EmcmfigmB 74 3 NH ON O O .3 939m m.N _ O _ _ O _ A _ _ ON O¢ OO Om OOH uogssgmsuul mag) 18d 75 HuH masonwmeoEETm-235280226; .mYvuafofiaxgozzo-H we Enbuogm GENES .mH 93me GeogoHEV SwamHgmB NH OH O HN m .N _ H O _ _ _ _ O 3 O I J ll '1 _ _ _ _ _ _ _ r H ON OHV OO OO OOH uogssgmsuml wag Jed 76 ¢H masonmbogcfigé-EwcmnobEéélHEEHctoncoHPHU-H O0 Esbumam @979ch NH OH _ $20.85: 5w:2m>a3 O .2 8:? m.N _ . 4 ON OHV OO OO OOH uoyssywsuml, mag) .Iad MICHIGAN STATE 933"???” M; AGRICULTURL' AL“?! N511" 7‘;;.~‘..‘£ DEPARTMENT OF 0925.231?! EAST LANSING, mums“ ART: 1 o ’57 {INCHES/‘3: SUITE UE‘~E!\.'Z?..SI‘§' UP AGRICIMURE A33 Au}?! <1 gsjmci DEPAITI IVZLI'IT OF C3 {1311331 RY EAST LANSING. MICHIGAN .4 7 4 7 2 3 4 1 IIIIIIIIIIIIIIII 3 1293 03 IIIIIIIIIIIII