[PAGE 1]
1221

TO: TSS/OC

1. Date of Obligatioa:N/A

2. Porpose of Project: To aupportstuly

3. Progress to Date: Gatissectory

4. Exptratioa Dase:/A

5. Prpject Monitor:

FROM: TSS/CD

Roora,

[PAGE 3]
Project HUUrRA 122

Purposet To develop a technique for-the-preparation and characterization of purified Heurokinin. Thls\_atudy ia clooely related to the vork of C

Initiated:- Sopteaber. 196O.

Contractor; as a granțee of. Cost; \$2,000.00

Status:.-Unknowm..

[PAGE 4]
Attached is an accounting of one of the ones they are waiting for.

[PAGE 5]
2 Kynographys twth drums of 6 and 12 in.

1 kyrograph mrith 6 in drum

2. Material purchased vitha Money Exchange \$203.09 \$1,103.09

Using an average rate ofthe following were purchàsed at the approximate-\$-value of

This is a true statenent of expenditures as reported to the fynd.

I ccrtify that corvlces or paterials nve a eattoraetecty received anl tho expenditures wero Jaeucrcd cn ortiotal tustneco.

O

122-4

[PAGE 6]
Cost Account . 125. 1002- 1102

Object Class .

192-5

(When Filled Ip)

[PAGE 7]
MEMORANDUM FOR: COMPTROLIER

ATTENTION : FIRANCE DIVISION

SUBJECT : MKULTRA, Subproject 122

Under the authority granted. in the momorandua dated 13 April 1953 from the DCI to the DO/A, end the extension of thia authority in subaequent momoranda, Subproject l20 has been approved and \$2,000of the over-all MULaRA project funds bos been obligated to covér the subproject's expenses.. This obligation of funds should be chargcd to A11otment 1525-1009-1902.

TSD/Research Brauch

CERTIFY THAT FUNDS ARE AVAILABLE OBUIGATION REFCINCE Ko. SHASGE O ALONENT  MES eY wot

Diatribution:

Orig & 2 - Aadrosses

1 - T80/nD

9 Stemptember 1960

[PAGE 8]
l-eel

[PAGE 9]
e e Bad soe  o Stc  e Stno se ns o t n ue et patt  etm der  e    e e t  e  t e a  te

coay t ty t y ts or Rre r ' re e re e rr s err e  er  o r

s ett o teoc at teae pas tsrt pez-mat as toate potued But an e        e  e    e  e e e Lee ses  o ee a r r e t  ere oe    e ce oer e ss yoeie gue aisotitireo s srsdaid geutsaco lot st litagea g ua aeed aut wcir pauteigo idtaoea aut n r r r Atesr r r cacr r aer r ser r r eer  tcco

e    er  es s   s  te Be      sn een cn s       nns

F Dt tt Sutt  e ot pecee stst t t  e set    t st  o eee se at

[PAGE 11]
19 Septeuber 1960

M4EORA5DUN FOR: TS CORD

I TsD/sudget Officer

SUEJXCT 44 KangaA, Cuboroject 122, Invoice 91 A11otoent 1125-1009-1902

1.. Invoico No. l is ettached covering the above subproject. Foyuent should be wéde as Tollous:

P 2. The check chould te foruscied to Chief, Tso/Research Erreh. taouh kso/iodsget Officer, .no later than Naudoy 3 Gotober ly60.

3. Thio is a final invoice. Hovever, since it is enticirated that taaitlonal Mods vill bo obligated for thia project, ths files should pot te closod. 4.

ChiofT60/Reseoreh Bronch

Distribution!

O01g 6 2 e Aressoo

[PAGE 13]
r  S  r. no ee se te ee e  ee  ta  t st t t e st tt

oatoe se o o pe ro see s  pco, e se rte S r r       s t    n  n

t   t t t  t a t    te B te    se     t  e t su    t  e se    eet s e oer e e pe       r  ass ee e oe pet  e r r ter t r  er er ar er eer er ir r  e

cott  rt ry t rts. r e r   r r r e      re

e a r t    e     t t e t   eto m  se  s  ae e e o pe  t e a ie ea D    to r  oe r str

SLONS

[PAGE 15]
tsTaA, cubprojec122, Invoieo f1
A11ovant 1025-1009-1902

l.  Involec Ko. 1 ia attuched covering the above cubproject. Poyoont shoua be wode cs tollovs:

Cashter'o check in the ecount of 2,002.(2 dreva on a local bank, lovablo to a

MERNNTm 2."fha chdck shoeld ba forutardel to Chiot, Tao/Reooarch Breach, through Ta/holget Cfficer, no lster thon Hoadsys 3 Catodor lç0O.

3.: Thio is a firal invoico. Rvever, sirce it is enttoireteá that esditiosal fuedo vill de ebllcted for this projeet, the filos should rot be elcsed.

Attotedi Trvotca à Coxtilicatiocs

Distriletiont Oric &. 2 " Actrosseo

1 - TSD/FASS

72 - TSD/XB

TSD A 19 Sept. (0)

199.12

19 Septosber 1960

VIA

K2ORAEOH4 FORI THS RCORD

SUBLCT

;2so/sulget offieer

206

[PAGE 16]
INVOICE

For Sorvices

\$2,000.00

CRRTIFICATTONS

(1) It is hereby certiried that this is Invoiea No. l. applying to Subproject lo. J2? of HaLaRA, thot pertoroence is satisfactoay, that services oro boing cecoaplichod in eccordance vith cutual egreenents, that a Gotailed. ogonda of the paymeuts and roceipts is on file in pso/ IB, thet this bill is just snd correct and that payuont thereof heo not yot been wade.

Chief, To0/Resoarch Brench

Dstet

(2) It is hèreby certtfied that this invotce applies to Subproject No. i22 of KuRa vhich vas duly epproved, epd that the project is boing carriedout 1n accordence vith the cenorandua of 13 Epril l953 from the ICI to the Dp/A, end tho extension of this authority in subsequent pemoranda.

Date!

Rescereh Director

O

102-12

[PAGE 17]
9 Septorber 1960

MMORANDUA FOR:COMPTROLIER

ATTRETION : Finenco Divipion

SUEJECT IKULTRA, Gubproject 122

Under tho cuthority gronted in tho secorantr dated 13 April 1953 froa tho Dl to the DD/A, cni tho extensiou of this quthority in subsequont resoranão, Subproject 122 has boen approved enl §2,000 o? tha over-all leVtst paojoct fuaãs has been odligated to cover tho svbprojeot'o oxponsos. Thio obligation of funãe ohould be charged to A1lotaont 1525-1009-1902.

Dato

Keseareh bireetor

122=13

APPROVED FOR ODLIOATTON OF FUOS:

[PAGE 18]
MEMORANDUM FOR: THE RECORD SUBJECT. : Project MKULTRA, Subproject 122

1. It is requested that Subproject 122 te approved to support Univin part the recearch project of ersity in the

2. Neurokinin is almost certeinly a major mediator in the human
"stress" reaction. Its nature and function are not well understooì.
Purification and chemical characterization are-necessary-prelinainary.
to exploring its biological function. thas until recently
been associated vith research program dealing vith
human atresa reactions at
Succesa, bA in the present study could support paterially project, MKULTRA, Subproject 61. has been briefed fully on this project h. Thia project vill te funded through th for security end cover purposes,
The accounting for funds expended shall confona to the eatablished
procedures of that organization. Title to sny permanent equipment shall remsin vith pniversity in lieu of overhead charges. 5. The total cost of this projeet for a period of one year is estimated to be \$2,000 as iniicated in the attached budget. Charges

[PAGE 19]
ehould bé pade againat A1lotment lmber 1525-1009-1902.,

6. No cleared or witting poreons are concerned with the

conduct of this project.

APFROVED FOR OBLIGATION OF FUNDS:

Date91171100

Attachament: Proposal

Distribution: Original Only

128-14

O

[PAGE 20]
MEMORANDUM TO:
SUBJECT: Prolect Pioposal entitled: Principal Investigutor; Th    crult All comments appear favorable, particrtotly thos who is a professor of pharmacology ot the It would appcar from his cyuluetion that this reseurch could constitute
fine scientific contribution. ns you moy know, has workod with
here ak and recently refurned to

My only comments are: 1) I do not feel personally àble to judge the signlficonce of the research except from a general point of view and, from that standpoint, it oppears to te xery Japortant; and 2) my only other reaction is the same as is the observation hu which questions the imamediate relevonce of this proposal to the fleld o If this observation can be raticnalized, then I would feel that the positive comments of the Directors and Advisers can lead only to recommendotion for approval to support this project.

August 15, 1960

Enc: 2 copies of Proposal

[PAGE 21]
188-15

May 1, 1960.

Dear Sirs:

I have onclosed a proposal for consideration by the for support for a series of studios conce med with Neurokinin, 5 substonce thot appears to be of great importance in tho pathological .mochanisms by which mon's relationship to his environment influences health.

Sincorely yours,

[PAGE 22]
Institution: Department of Pharmacology, University of

The contral norvous syster evokos, Integrotes, and modulates tho body's adaptivo rosponses to the cnvironment. These responses, necessary to survival when oppropriate, are implicated in tho etiology ond course of discase of mony categories when inoppropriote. It is well established that reactivity to noxious stimulation can bo modulated by tho contralinorvous system through control of the timing and amount of-adrenal cortical stimulatigg hormonę (-AC TH)from the pituitary gland. Recent experiments at the have shown that another major route for central nervous systom modulation of the inflammatory rosponse exists. This latter route Iis through vosomotor control and through modification of the magnitude of local,neuronal roflexos. A potent vasodilator substonce is implicated in these reactions The applicant has boen ono of tho yho has defined

this biochomical substance as nourokinin. Neurokinin apparontly is a vasodilator polypeptido formed by the splitting of a protoin (globulin) prosont in extracellulo r fluid. This splitting is due to the action of a protoclytic onzyme (ncurokinin forning enzyve), posslbly an Intrancuronal protouso. The some enzyme has been ebserved

PROPOSAL

199.15

[PAGE 23]
in the cerebrospinal fluid and in perfusato of the cerebral vontriclo during stimulotion of the brain, thus giving à source of the material for study. A crude preporation of powdered neurok kinin hos been propared by incubation of globulinwith cerebrospinal fluid collected from pationts with schizophrenia, which contçinplarge omounts of neurokinin-forming enzyme, NT
Tho applicaticn hos worked with and in the
definitien-sf this agent and in the proparation of cfude neurokinin.

e) Immediate Cbjectives.

To prepare and chāräctērize highly purified neurokinin by chromatographic

chemical methods.

f) Long Range Objóctives

To defino tho pharmacologic properties, and contribute to understanding

of the physiological and pathological significance of nevrokinin.

Il, Methods

: a) Obtoin nourokinin forming enzyme from humon cerebrospinal fluid and

from porfusate of cerebral ventricles of laborètory onimals during stimulgtion of the brain.

: b) freporotion of crudo neurokinin by incubation of cerebrospinol fluid

with globulin, precipitation, and extraction with alcohol.

c) Purification of tho crude preporation by paper and column chroma-

tographic seporotión of the compononts in the crudo preparation.

d) Pharmacologic studios to Identify the active fraction

a. isolated rat uterus, rat duodonum bioossay

and ossoy of depressor action on tho blood pressure.

O

132-15

2-

[PAGE 24]
e) Pharmacoiogre charozteñiectian of the propemes of purifi. neurokinin.

I) Electroencephalogrophic chonges induced by intraventricular ond

intcrcarotid injection in the cat.

2) Behovioral chonges induced by introventriculor injections of

neurokinin into unanosthetized cats with implonted intraventricular connulae.

11l. It is expected thot these studies will require one yeor to complete.

Budget

a) Apparatus to augment :equipment ovailable in the

I) Stereotoxic. (Horsely Clark) cpparatus for perfusion of cerebrol

ventricies and for placement-of intraventricular connuloe \$900.00

2) Perfusion pump 200.00

3) Small polyethylenc Tubing 50.00

4) Rocording kimograph for smooth muscle bioossoy 250.00

5) Miscellonoous equipmont, manometor,clamps, 100.00 cannulaè

b) Drugs ond supplies for clectrophoresis and for phomacologic studies 300.00

c) Animals, animal food, and coges 200.00
TOTAL\$2,000.00

-3-

[PAGE 25]
is o tax exempt orgonization of tho Tho cuthor is engaged in full time reseorch ond teaching. He hos worked as a professionoj Foryeors and has worked specifically with vasodilator. poly peptides with He hos recently completed a followship with working with neurokinin and bocoming acquainted with the point of view of tht program.

This project has-not been undertaken previously and no support hos been

orr is being requested of other foundo fi ons.

[PAGE 26]
The. contral nervous system evokes, integrates, and modulates the body's adaptive rosponses to the environment. These responsos, necossory to. survival when appropriote, ore implicated in the etiology and course of discose of mony cotegories whon inoppropriafo." It Is well established that reactivily to noxious stimulation can be modulated by tho centrol nervous system through control of the timing and omount of adrenal cortical stimulgtigg hormonə (:AC TH)from the pituitory glond. Recent experimonts at the hae shown that another major route for central nervous system modulation of the inflammatory response exists. This lotter route is through vosomotor control and through modificalion of the magnitude of local,neuronal reflexes. A potent vasodilator substance is implicoted in these reactions,

Thē applicant has been one of the investigdors who has defined this biochemical substanco as nevrokinin. Nourokinin opparently is a vosodilator polypeptido formed by tho splitting of a protoin (globulin) prosent in extrocellula r fluid; This splitting is due to the action of a prof:oclytic enzyme (ncurokinin forming onzyme), possibly an Intraneuronal protcaso. Tho same enzyme has been obsorved

[PAGE 27]
in tho cerebrospinal fluid and in perfusato of the cerobral ventricle during stimulotion of the brain, thus giving a sourco of the material for study, A crude preparation of powdered neurol kinin hos been prepared by incubation of globulin with cerebrospinal fluid collected from potients with schizophrenio, which contdsșlorge,omounts of neurckinin-fonming enzyme. The opplicon hos worked witt and in the definitien-of this agent end in the proparation of crude neurokinin.
e) Immcdiate Cbjectives.

To prepore ond charocterize highly purified neurokinin by chromatogrophic chomical methods.

To defino tho pharmacologic properties, and contribute to understanding of the physiological and pathologicol significance of nevrokinin.

II. Methods

a) Obtuin neurokinin forming onzyme from humon cerebrospinal fluid ond
from porfusato of cerebral ventriclos of laborctory onimals during stimulation of the brain. b) Preparation of crude nourokinin by incubation of cerebrospinol ftuid
with globulin, precipitation, and extraction with alcohol. c) ¿urificatlon of tho crudo preparation by poper and column chroma
tographic soparation of tho compononts in the crudo preparation. d) Phermocologic studios to Identily tho active fraction a. isolatod rat uterus, rat duodonum bioassay and ossay of dopressor action on the blood prossure.

122-1

2-

[PAGE 28]
O

e) Phormocologre ·éharacteiixetian of the propemes of purifi nevrokinin.

I) Electroencephalogrophic changes induced by intraventricular and

TO Intçrcorotid injection in the cot.

2) Behavioral chonges induced by introvenfriculor injections of

neurokinin into unanosthetized cots with implonted intraventricular cannulae.

IlI. It is expected that these studies will require one year to complete.

IV. Budget

a) Apparatus to augment equipment available in the

I) Sterootoxic (Horsely Clark) opporatus for perfusion of cerebral

ventricles and for plccement of iniroventricular connulae \$900.00

2) Perfusion pump 200.00

3) Small polyethylene Tubing 50.00

4) Recording kimogroph for smooth muscle bioassay 250.00

5) Miscellanoous oquipmont,monometer, claups, 100.00 cannulcè

b) Drugs and supplies for electrophoresis and for phormacologic studies 300.00

c) Animals, animal food, and cages 200.00
TOTAL2,000.00

-3-

122.5

[PAGE 29]
V. The outhor is is or tox exempt orgçnization of the State The outhor is engoged in full time reseorch and teaching. He has worked as
a professiono years and. bas worked specifically with vasodilator
poly peptides with Ho has recently completed
fellowship with working with neurokinin
and bocoming ocquainted with tho point of view of progrom.

VI and VII :

This project has not been undertaken previously and no support has been

cr: Is being requested of other foundafi ons.

[PAGE 30]
26 Angust 1960

LENORANDUM FOR: THE RECORD
SUBJECT Appçoel of Dr roposal
ATTENDANCE Gottliel

SUHARY: Request for \$2000.00 to provide equipment end.. supplies foz study was approved. Neurokinin is presumed to be a wsjor mediator in the "stress" reaction. Its nature and function are not vell understood. Purification and chemical characterization is a necessery prelininary to exploring its biological function. This proposal has received the strong endorsement of scaxt

The fudine necharism vill be th Title to permanent equipuent 3. A nsme chcck of proved negative. He will reoain unwitting of true Spohsor

Distribution: Orig & 3 - Addressee (TSD/RB)

Rac'a. Pvd. 9/g 917.60 2751.+ dicr6d

[PAGE 31]
MEMORANDUM FOR: THE RECORD

Mechanisn,1ts"611/1n7nd
Troa his partictpation in the old I assured
him, in enswer to his question, that erranpgemcnts could be nade
to approach grantoes to undertake missions.Further
if his diyision needed a funding mechanisa with sound cover to
promote any  that could be fitted into the cover, the was available to help out.

2. Although saw no immediate neéd, he wos gemvinely enthusiastic and wished to meke division brass aware of the possibilities.

Distribution: Orig & 3 - Addressee (TSD/RB)

Rec'd. Fwd. 1119 8 84 80 1k0

Ifor file

k

22

23 Auguat 1960

182-17