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ANNUAL PAOGRESS REPORT

H. F. Frassr

#s

and Harris Isbell

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NIiH Aadiction Rasearch Center PrS hospital Lexiagton,nrntuchy

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It is alaost irposstble to suarize tha work of the NIMH: Addiction Research Center for a year In a. short presentation, and this is becoming steadtly mors difficult es our activitics Incraase in size and scope. The progren ncw encompasses work in cllnicel pharzacology of analgesics, clinlcal studies on psychpsonisetics, clinical psychopharracological Investigations, blochemiasl experiments ranging from clinical encocrinoíegy to drug retabolism, complex neuropharmacological and noyrephystologfeal invcsttgatlons, and psychepharnacologiàl studiss in anirals. In each or these areas a qreot anount of wors has bsen corplsted curing tho year, so obviously 1 can do no more than touch vany sketchtly cn a few of the hichiights of work with thich l have daen directly involved and which, I hope, will de of Interest to this group.

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As you know, Dr. Frasor has found that substitution of new analgesics for rorphine in audicted pationts for a- puriod of 24 hours is a sensitive and uscful test of the abilily of a drug to suppress abstinsncs fron norphine. Substitutlon for 24 hours has several advantagus: Unly minimal anounts of new analgesics, the toxlcity of which is frequontly unknoun In ran, are required. Very little tolerunce and cependance are lost because of ths short perlod of substitatlon. Patients do not bacoms too uncoafortable during the lest period, so that experinents can de repeated veekiy. 24-Hour substttution has reflected relatively well the results with the classic l0-cay substitutlon cethod. Tha 2h-hour procedure coes not, of.courss, reflect the Intenslty of abstinence foliowing withdrakal of the test crug after subatitutlon for morphine.

Becsuse of our interest in studying cissociatlon in crug elfects (s.Q., "eupheric" patsncy as contrasteq with potuncy in suppressing abstinence, or intensity.of abstinencu on witheranai), ve ere altepting to develop care quantitative ways.of studying these phononena.

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New Fethods for Cetermining Acdictiveness of Analgesics.

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During the year, Dr. Fraser has collected hourly point acores obtalned durtag substitution of 100, 50,20, 10 ana 0 per cent of the pitlents' accustomed doss of morphine in a 24 hour period. The procedure used was as followsi Fatients were stabllized on 60 ag. of morphine four times dallyi. at 4 p.m. on day prior to the test, the last regular dose of sorphine was givonz at l0 p.m. of the same night, 6 a.m. end 10 a.m. of the rollowing morning patient received a subcutanvous cose of an "unknown" msdication (actually rorphine in coses of 60, 30, 12, 6 ng. or a placebo). U'sual observations fur intensity of abstinence were made from é a.n. to s prms on the test day (lhth to the 2hth hour of abstincnce), and the houriy point scoras calculated according to the.Hinmelsbach syetaa (1). prea under the curvas was, calculeted by the method of winter end Flataker (2), thus converting all the data on a perticular patlent for a particular day to one figure, called "point-hours." In all, we have collected data on 39 putients after substitutien of placsbo, on 2i after substitution of the regular sose of morphine, and on 9-after substitution of 10, 23, ana 50 per cont ef the accustomsd dosa.of morphino.. Results sre showm in Figure l. .Tha brackuts at éach point indicate tus standard errors above and h:low the mean for that particular point. Obvlously the data fors a very pronising but incorplets dosoaffect curve.

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It is our hope that this curve can be definsd so voll by adding other patlents at the polnts alreacy studled, by stadying other coses, and determining llnits of varlations for dirferent groups of men, that we can sake reasonably accurate estimations of the dose of new analgeslcs which would be: equivalent to e standard emount of morphins in supprsssing abstinenca. We will have to study the suppressiva potency of two or sore dosa levels of new analgcsics and corpare the dose effect curves. so obtained with the new' drug with that of the standard morphine curve. Since we can use a patiant only once a week and for a lizited nwber of trials, this is abviously going to be a long-ranga project. ve do not expect to use it with all cospounds subsitted to us. Only those drugs of seru theoretical interest will be stucied in this way-

We also hope to develcp a quantitative systea for evalusting. "suphoregenie" petoncy. Tha initial effort in this line is. balng carriad out by dsternining the porcantage of patisnts who moxe positivz ldentirications that tha subjuctiva effacts of "unknown" drugs - In realitys.various doses of rorphine  reserble those of aplates or sons other drug vhich secicts rosarj es plcasant.This is an all-or-nono encpoint and the rosults will da subjectod to prosit analysts. As yet a sufficlent number of patients has not been studied to deterains the. potonttsl va-lue of the procadure.

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PinaLly, we aro studying the efficacy of a, short "direct addictlon" procadure in cetermining the intensity of asstinenco after cirect withdrawal. Paticnts will receive the new drvg for a period. of 19-20 deys with the dose being clevated as rapidly as tolerance peraits end observatlons for Intonsity.- of abstinenca carrled out for l0 daye after abrupt discentinuation of the pedicatlon. Calculations forIntensity of abatinance will be converted to area flgures ("point-days" in this case) and conpared vith ths intensity sfter sorphine or codeine. Currently saven drugs, including NIH-7519,and Nll-7525, are boing studied by this nethod.

At the tino this report was written studies on all the new analgsalcs subaitted to us sincs Parch l936 ars still incomplete so that no final cvaluation or any cf these drugs is possible. Howevar, sonz.of the results are of considarable interest so the following prelintnary reports are beiag made.

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lata ethane sulfonate (ai. This cospound Is a. congener of neperidine, or demerol, and
poosesses a norphine-lika spectrun of pharracological activity. NIh-7590 1s spproximately twice as active as. morphine as an "ancigesic" in mice. it ls about 18. times es potont as morphine In suppressing abstinencs in tha monkey. The drug is an effectiva analgesic in man, and is probably as potent as morphinu in this respect. nssnect. • eunteetiva. effecta In 15 to 20 mg. of HH-7590 inducad subjeetiva. eflecta in
10 nontolerant former acdicts which were identifled as rcssmb
ling those of heroln or sarphine by the tafority of the subjects. Four patients Wo vere actively audicted to and stabillzed
on 2L0 mg. of porphine sulfata daily received NIí-7590 io tares 2h-hour tests for suppresslon of abatinencs. ln the first tost 12 mg. of the drug wors given every six hours (total of 36 ag.)s in the second, lo rg. were given every thres hours (total or S0 mg-); and in the third, 15 nle gvery thres bours (total or 75 sgs). The rosults are shown Ia Figure 2. The average score wita lZ ng. every sik aours vas l09 "point-hours" which corresponds approxieately to the level of abstinence sean vhen 20 por cant of the doss of norphina is substltuted (36 rs, of

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corphine in threc doseali the averaje acore with 10 mn. every thres hours was 79 "point-hours;" the averoge score with 15 ng. of:HiH-7590 cvery thres hours was 58 "point-hours," which corresponds to a point on the norphine curve beloy tha laval seen after substitution of 5J per cont of the accustozsu duse.. of morphine.

Because of the nall nurber of subjects, ve have not mads the statistical calcilatfons for scope, parailelisa, and
comparative potencies on these curves. This will be done when more data have hoen accunulated. As you see In Figure 2, we have drasa the curve for Nil-7590 through the two polats
odtained with 10 and 15 ng. every three hours and have
disregardad the point obtained after 12 ng. every alx hours. Ths reeson Is that In the first tost (12 mg. every six hours) it was evldent froa. inspection of the data thet Nil-7590 bas suppressing abstineacs for only thrcs hours, or, in ths words of the addiets, tas a "quick-burning" drug of short length or actlon. The six-hour sczecule, therafore, çives. an iapresston of suppresslvs petency which is somawhat too low. By interpolation fros the curves as crawn, 0.5 ngr of Nin-759t is cstiatea to te eeat to l . of cerphins. This is caly as ostirate which ray ba rovised aa rore data becore available and statistical calcuictions cone. It is, towever, svlcant that Nili-7590 is. very effeetiva. In suppressing abstinenes and ts at Icast as potent.es porphins in this rospect. It Is, of courso, not nearly as potent In ran as in the ronkey.
dn\~

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L-3-Hydroxy-li-phenacylrorphinan ncthans sulronato (N1H-7525, Ro k-0206/1).

Thls conpound Is of considerable interest becauss, although it Is approximataly 30 tiaes as potent as norphine in analgesic tests in mice,. it is only one-fifth as potent as corphins in suppressing abstinence in the monkey, It, therefore, secaed possible that a considurable dissociation between analgesic effect nnd abliity to suppress abstinence might have bucn echieved in this drug.

In 7 nontolarant forzer césicts, doses of 2o ag. of HIn-7525.aubcutaneously Induced subjeetive effecta pesoabling those e5 aorphine in 13 of 16 trisls. Nll-7525 1s probably 5-10 tines as potent as zorphine as an suphoriant.

Three patients who were addlcted to and stabilized on 240 mg. of morphine -sulfats daily received 3 ng- (one patient, total dose 9 rg.) or 5 mg. (2 patients, total doss 15 ng-) or NIl-7525 subcutanoously every.six hours turlng. a 2-hewr substitution test. The "point-hour" scores on the 3 patients were 73. (9 mg. total. dose), 18 (15 mo-) and 59 (15 ng.). Thesz scores are equivalent to average suppression by Ea, ll2 and II2 mg..of corphine during the sane period of tise, Thas, as

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a rough prebialnary esttmate, NH-7525 scems to be 7-5 ilmes aseffective as scrphine in suppressing abstinenca in man. Further tests are currently underway in order to doteraina suppressive potency sore exsctiy.

One ahort direct acdiction test has been complcted. Cna
nontolerant forner morphine addict reccived NIn-7525 In doses
Inarcasing frog 1 ag, to 7 sg. subcutansousiy over the coursa
of. 1o dave.Marked aorodine-like behavioral and sabjectivs
chanoes were observed, includipg puplllary constriction, 2
denresslon of resgiratory rate, pausca, and vorltlng. The
oatlant had toxic gffects with dosce of 5 nç. four tloes daily.
On tha l7th day of addiction, 5 mg. of calorphins subcutaneously
prscipltated mlld but dsfinite syoptons of abstinence. Fotloving
sbrvot withdraval of klli-752s, aild tut definite abstinence
appearee an the third day and porsisted through the sevonth day. The results are cefficlent for a tentotive statanant that
NiR-7sas has adiictive properties. Asssssnent of the doçrea of seiictivincss must avait cosplatioa of adsittozar aspirlnonts.

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Dr. Everett Pay at NIll and studied by Dr. Nathan D. Eddy. It 1s nearly l0 timas as effective as morphine as an.analgcsic.. In mice. Cespits this, It is much less effective than morphine in suppressing abstinence in monkeys (Eose equivalent to 3 ng. of norphine Is approximately 17 ag.). Preliminary results indlcats that Nil-7519 is a petent analgesic in man. Since a dissoclatlon batween analgeaic and,abstiapnca-suppressive potencies night be presant, hurnn experinents were undertaken.

In 8 nontolerant forzer porphine addicts, j-4 eg. of Nll-7519subcutaneously laduced carkad rorphins-like subjective and bohavioral efrects in 12 of 17 trlals. me drug is probably 5-l0 times as potent as morphine as an euphorient.

Four patients who were sddicted to and stabilized on ZhO ag. of porphine sulfate cally roceived 2 ag. (total of 6 ng. in substitution perioo,4 pattents) or 3 ag. (total of 9 ao.. 3 patlents) every six hours in 2h-hour eubstitution tests. Abstinence was suppressed very efrectively Ly these Cosus.

dl-2t-Hycroxy-2,9-di:nthyl--ohenethyl

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The average "point-hour" score in 4 patients recciving 2 ma. avory six hours vas 76. Ths average "point-hour" score in 3 patients after 3 ag. overy six hours tas 83. NIk-7519 is probably at least5-l0 tirea as potent as sorphins la suppressing abstinenco In can.

Tho patfents have. undergone. short direct addiction experloants. The dose of NiH-7519 was increised.Irom 5 ag. to 24-36 mgi datly over the course of 19 to 24 days. Farhed sorpninz-llke behavioral changes wers observed In these patients. - who also rsported intense morphins-rihe cubjective effects. Both patients were soecuáat toxic on-this dosags schsdule. 5 ag. or nalorphine precipitated aild abstinence in both inotances. Follcwing vilhdruzzl both patients had dafiaite, though nilu, abstinonte.

NIk-7519 definlteiy 1s an eddictive drug. Hovever, lire Nll-7525, the ougres of addietivensss cannot to assessed preclsoly uatil other experinents which ere underway ars cosplcted. Of greatost interest to us at the ricrent ere the qrantlsattve discrepanctes betwsen suppreasive potencies in ran snd the tonkesy, snd tãe cflencss of ebstiaaeu claerved aftir ehert elraet eddiction in zas. Cvalcatien ee lie lalter poiat wilk, or ceurse, depend on a cosparison of tho intensity cf absiinenes after these Crugs vith that alter adiiniatratien uf norphine for e cpuparable perlod of tin.

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You wilirecatl that, In curpetients, nornorphina is, In single dose, far less potent then sorphinc. Yet when sxail doses (lo mg. subcutansously) are given, sedativo.eifects of normorphine accumulate to such an extent that the dosage of normorphins cunñot ba elsvated as repidly or to as great a degrea as ls the case with norphine. Following withdrawal of nornorphine, abstinence Las much nilder in Intensity than was abstinence froa zorphine. For this roason, a cocparison of the ratabollc fate and rates eß. usinary excretben-oi: sornorphina and morphine in raa tas undertaken by our blocheaists. Frs. Jewll Sican and Dr. A. J. Eisenmn.

The nethod uscd vas a sodification ef that of Axeirod and Cochin (3). it dspends on exirection of the norsorphina fron urine at pH 9.3 into a mixture of 20 per cent amyi alcohol in sthylene uichlorids folicued by re-oxtraction or the norsorphins fato aquoous sqluiioa waiea Oeobk lCa Elter wich a blue color is daveloped by silicopolybuatc reagent as dascribed, by Fujinoto, et ai. (lo).. Horphins tas estlated by tho method oi Fujinoto (4). The. method ror norporphine givss consistent and reproductdle recoveries-of added norsorphino. Tha material being cutereined hes boen Idantirfad vith a hich dogreo of certainty as being uuthentia narmerphine by asans of ppper chrocstography and ceuster-curront otstrtsution.

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Hetabollc Fate of lorrorphine

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As ls true of morphins, noreorphine is excreled in two. forns free" or readily extractable and "bound,"or extractable oniy after appiicatlon of some hydrolytic proeedure such as heating with strong acid or incubating with beta-glucuronicasc. mere is, however, a mrked difference in the degree of "binding" of noraorphine as compared with sorphine. You vill notice in: the following alida (Table 1) that "froe" morphine accountes for only Il.h per cent of the: total rorphine recoverable fron tha urina of 3 patlents who recsived 70 ng. of the drug, whsreas "free" normorphine accounted for 5i per ceat of tho totet excreteu. Thesu data confirn those prevlously reported on othar pellenta.

Not only is the degree of binuing or normorphiae far leos than is the case with morphins, but normorphine saess to be conjugated dirferently. The priacipal conjugata of sorphins is inuwn to be the phenolie giucuronide. mhen urine containing "bound" norphine is incutated with h00 units of beta-glucurenicass par cc. of urine at pN 6i2 for 65 to 70 hours, hydrolysis of "bounl" morphins is neariy as cooplsta as vith. urastic acid tydrolysic. On the othar hand, incubation of urins containing "bound" nornoryhido vith bota-glucurontcass uncer the sana conaitlons causes no or very littla hyurolysis of tho "bsand" merrorphins. These cata suggest that In ran nornorphine is not conjagatad as a phenelic glucurenics.

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Current work In thls fieid consists of efforts to detor aine the nature or "bouau" ncroorphine and to isolato it. The seaning of these. differcnças between the metabollsn of morphine and normorphine are still obscure. Cnc hypothesls might bo franed as followst The "free" forns of norsorphine and sorphino are the active forns of thess drugs; norsorphins is Inherentiy a weaker drug than norphinet unier conditlons of chranic adainlatration, less noroorphine Is bound: so that core frce and, presurabiy, active crug accuulates in brain than ls the cass with rorphine. Obviously this hyothesis Can only be tested in anirals and, before ve can fast ft, we seet find an aniral that netabollsos porphène cad nornor;hine in the sáso vey es doso rin.

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Yeshave long bean. interested in and have carried out a great rany Investications cecling vith drugs that inuucs poyclosss. Interest in thse coxpounus steru, in part, fren the fact that tho of the drugs controlled by the narcotic laws  cocaina Lnd rarlhuena  are, in e sensa, psyuhotoninetics ane, in part, frun our interest in the relatfonship. of thesa materlsis to lerger probicas ef mantal ciseeeo. As 2 pou know, a nurtder of hypothcaes ravo been forzulated wiich relate paycesorisetie. effects to s ceflclency of, oe. to an excees of the to-celled neurohurors, epincphrine, norupinaphrins, ecetylchotine and scretenin (Deficiency or ezcess is thoucht of tn tcrrs ef pharzacolecioal effect rather. than concortratler). Greutest erphasis has bosn piacod on serotonin.

Curing the yzarr we cospleted an examination of a nunber of cengeners or uicthylaaida of lyssrgic ucid, whien vara pade evalleble to us through the kindnoss of Dr. R. Bircher of the Sanlos Corpany. The potancy of. thesa druçs In blocking serotonin-induced contractions. is corpared with, thetr paychotonutie potoncics la Table 2.. Unfortunately, tine doss rot perait a coscripticn of the sethods by whlch the psychotomirtic potencieo ware datsrsined, so ths figures pust bs takon on faith. You will notice that high potency ss an antisorotonin

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Paychotoninetic Druus

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is not. nacessarlly correlated with hlch potency as a poychosocinetic. Ct-ll3, ret-6l, and TLA-74 are all sor: potant than Lsb as sarotoain Llcciars yt arr not nuarly es pcteat. as pevchotorinatics. Cn the otter lan, there Is re Instasa. o2 a. cespouad in thls gruup tiat is a petcnt peycotonirctic that is not aisc a potent antlscrotonla. Tustia dta, vhile not favoring thesecotoafe-dafielency hypothcsls of tho L50 psyclosis, Co not áisprevs it.

Another interesting finding duriag the year was the detection of pstent hypnotic cffocts in cuxpornda petatnd to Lso (Flgurs 3). Te erugs In vilch vs bavoe datuated this sort o5 uctivity ars all couonnas In cátoh the auid aafia graup af LsD das been raplacod dy alkyí ani hydroxyl qoaps. Tas corpouncs ie have studied ars apociavia ead ahyéroaçroclavias (taheda Co., Japan) and a elosely alailur copound knom as Liliy 2iiR. Cha lilly drug Is tho cost potent hypnatic of the thre. so va vill presaat aaiy tho data on that crug. la antrals both sgroelavtnt end Lftly 23itl bete excitant, wapoas dinydroagroclavine sis a dapressant cruj. In our phttants, boeover,  tso. co efe not izec: ttsor 3 pe croaslntos and eroer. corparisen or nypnstis artvr 2 and  ng. of Li11y (eur 50: scobaratial ls grscontet in Tiola 3.

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The rethod used-in evaltating hypnosis wais as follavst Patlents cntered the ward the night before and slspt throuch the ntght as.usual. Cn the foliowing corning thoy ware given, In randomized "doubla-biind" fashlon, 2 or h ag. of V-A-2i, 100:or 200 sg. of sooiun secobarbital, or a placebo. Nino. petients received all five trcatmonts. Drugs wore given with tha patlents fasting end coffec vas not peraltted. Patlents were obsarved at half-hour intorvats froa 8 a.n. to lppsm. to deterains if they ware aslcop. Lookiag at tha toble It iz evicunt Uhat both V-a-2i and sccabardital caused a sigaificant increase in slcep eT corpercd vith placebo. Tho greatest Incresse was in the hours fron 9 a.p. to noon. là isaloo evident that, althouch no cloar-cut dase effect vas obtained, that V-A-21 is roughly 50 tiros as potent as socolarbitalas e bypaotle uader these conditlons.

ye hevs excorimants in proçress in which we are attsipting to hlock the LsO psychosis. by conconitant adainistratlon of Veé-2l.Althcuch the experinant is not coaplato, It is airaady evident that V-a-2i doss not attennate ths LSD psthosls but antualiy ealanccs It.

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I see that I have used all the thus talking about soma of the clinical aspects of our progran and no thiaa is left for a discusslon of the iaportant and interesting tasle rescarches carried on by our ncurophysiological and psyciological sactisns. I hope that ot the next meetiag tr. Wikier and his co-korkers can cover theso aspacts of our work for, you.

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l. Hiemelsbach, C. K.t Studies of certain addiction characteristics or (a) Diaydronorphine ("Paracorphan")s (b) Dihydrodesoxymorphine-D ("cesororphine"); (c) DIhysrodegoxycodeino-D, ("cesocodeIno"); and (c) Nathyldihydromorphinono ("hetopon").. J.Pharoacol. & Exper. Therap.,67: 239-259 (0ct.) 1939.

2 Winter, C. A and Flataker, Li: Studles on heptasone (6-orphotino-,h-cighenyl-3-heptanor hyuiroehlosge) in comparison with other annlcesle erugs. d. Paazvacole à Lnper. Therap: 305-317 Gar-1 PE0.

3. Axalrod, J. ane Coctia, J. The lahisltory actica of nalorphine on-the enzymatic k-desethylatioa of narcotia druga. J. Pharracoi. à. Expes- Therap 121s 107-112. (Scpt.) 1957.

4 Fujimato, J. Ne, kay, 2. L. end Hine, C. Het A Fapia method Sos ths gstiration of tovghine. Ji Lab. 6 Cllne Pad., h:627-635 (Cct.) 195h.

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LEGEND FOR FIGURE 1.

Figure i. Relationship of dose of rorphins to intensity of abstinsnca in 2-hour substitutions.

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LOG OF DOSE O.5 0.75 Mgm/24 HOURS PER CENT DAILY DOSE

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LEGEND FOR FIGURE 2.

Figure 2. Suppression of abstinsnce by Ethyl -phenyl -E[3-(phenylanino)-propyl)-l-piperlcias carboxyiate ethane sulfenate (WIN 14,398, NIH-7590),

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LEGDND FOR FIGURE 3-

Fioure 1.. Structurai foraatae of congeners of LSD With hypnotic efrects.

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LEGSND FOR TABLE, 1.

Tabis l.  Comparison of excretion and conjagmt-ion of morphine ana normorphino after administration cf 70 s3- or both drugs.

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FIGURES ARE AVERAGES OF DATA ON THREE SUBJECTS. RANGE SHOWN IN PARENTHESES.

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LEGEND FOT TABLE 2.

Table 2. Comparfson ef poychotonimetie and antiserotonin potencies of congeners of LSD.

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Table 3. Comparison of hypnotic effects of V-A-21

and secobarbital.

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\* \* HIGHLY SIGNIFICANT \*SIGNIFICANT NS NOT SIGNIFICANT

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\* FIGURES ARE MEANS OF OBSERVATIONS ON 5 SUBJECTS \*\* HIGHLY SIGNIFICANT

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