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ART6-0133
International Research and Development Corporation
_r---335 |
SPONSOR: cowpounD: SUBJECT:
3M Conpany
Fluorad Fluorochemtcal Surfactant FC-95
90-Day Study.
Subacute
Rhesus
Monkey
Toxicity
EVidcweinPrI.estGobfndtentahnadl, Ph.D. Directordf Research
Collaborators:
D.DiC.recJtesosrupo,f PRhe.sDe.a,rcAhssociate
R. G. and
GDeiirle,ctoDr.V.oMf.,PatVihcoelogPyresident
J.ofS. LNaerhgreinAgn,inaPlh.DT.o,xicDoilroegcytor
Date: January 2, 1979
:
137-087
!
002070
International Research and Development Corporation
TABLE OF CONTENTS
To
SYMOPSLS
te
: eee
eee,
Page 1
TL Compound ot vena... 3
IIL Clinical Studies + vv ovens e tne. 4
A MEEhods Lue. 4
1. General Procedure + +. i.......... 4
2. Compound Administration . . .. Lo. ...... 4
3. OBSETVARIONS + + vai hai.aa.a.... 5
4. Clinical Laboratory Tests . . . ov .ou oon. 3
a. bo
BHeml atoloo gy c .soh eoa.e.a.im .a.ii .i.as .a..t ....r ....y ...
53
dc.ooStUanttniasltyisciasl .An.al.ys.is... ..L.L.................... &6
Bu ReSULES ov tna. 6
1. General Behavior, Appearance and Survival . . ... 6
2. BodyWelghts vs uiuaiaaa..a. 7
3. Laboratory Tests +s. a a .aaaiaa.... 7
2b.o HBemai toloo gy c . . +h .o4e ssiimui i aii s ua. t aa. r aa. y .. 77
co Urinalysis oo ..i......i.l...l. 7
IV. Pathological Studies + vv eit ut iuu ante. 8
A MeEhods Lui... 8
I. Gross Pathology ess aui........ 8
2. HISto +P vsA vst sihto etlaong iay. 8
Bu ReSuLES ov vivant. 9
1. Gross Pathology and Organ Weights . . ....... 9
2. Histo +p+va est uahaioialaioi.g..y. 9
Table Yo.
1. IndivBioddyWueiaghlts + vv outa... 10 2. Individual Hematological Values . +... ...... Il
137-087
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International Research and Development Corporation
TABLE (COonFtinueCd)ONTENTS
Table No. Cont.
Page
3. Individual Biochemical Values + +... ........ 1
4. Individual Urinalysis Values + + ov uu... 13
5. Gross Necropsy Observations of Deaths . . . ...... 14-15
6. Absolute and Relative Organ Welghts . . . ....... 16-17
7. Microscopic Observations . +. uu... Lo... . 13-2]
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I. srvopsts Fluorad Fluorochemical Surfactant FC-95 was adninistered to rhesus
monkeys, fn aqueous suspension at dosage levels of 10, 30, 100 and 300 n3/ig/day for 20 days. Tvo male and two female monkeys were initiated at each dosage level and also in a control group. The control group uas treated only with distilled water. The monkeys wers observa twice daily for general physical appearance and behavior and pharnacotoxic signs. ad weights vere recoric! weekly. Hematological, biochemical and urine studies were conducted only fn the control period. The study was terminated after 20 days because of the early deaths of the monkeys fn all treatnent groups.
The monkeys treated with 300 mg/kg/day died betveen the Znd and 4th days after the initiation of the study. From the first through the second day, the following signs of toxicity were observed: anorexia, decreased activity (from slight to severe before death), emesis with some diarrhea, body stiffening, general body trembling and tuitching, weakness, convulsions and prostration.
The aduinistration of 100 mg/kg/day of the test compound led to death of all monkeys between the 3rd and Sth day of study. The monkeys treated ac 30 mg/kg/day died between the 7th and 10th day and those treated at 10 mg/kg/day died. between the llth and 20th day of study. At each of these dosage levels the toxic symptoms were the same as for the monkeys at the 300-ng/kg/day dosage level. However, the time of appearance of the toxic sysptons vas related to dosage level.
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International Research and Development Corporation
rage 2 ALL deaths were considered as resulting from compound effect. A vellowish-brown liver coloration noted at necropsy in several monkeys rom the 100 and 300 mg/kg/day levels was the only gross finding which was considered compound-related. Yo histomorphologic basis for this finding was observed. Other gross and microscopic lesions wees of an agonal nature or were those "hich ar: commonly seen in urtrsared monkeys. Organ weights of treated monkeys were considered within nor al lates.
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1. cowpounp
The compound vas Teceived from MM Company, Sait Paul, Minesota
on October 24, 1977 as shown below:
Label
Description
FFCl-u9o5rad34 FStloucrkochNeo.nic9a8l-02S0u7r-f0ac1t0a3n-t7 white powder
Lot 310 Yet wt. 5 ids. 2,2 kg
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IIL cLINTCAL STuptes
A. ETHODS:
1. General Procedure:
Ten male (veighing from (weighing from 2.75 to 4.10 kg)
3.05 to 3.80 kg) and ten female rhesus monkeys obtained from Petaats
Taports, monkeys
Port were
Washington, New York housed Individually tn
were initiated hanging wire
into nash
this study, "squeesemtvpe"
The
cages Foon.
and maintained in a taspetacaves, twidtioe and Mirt-sontralisd Purins onkey Chowd was fad tulce each day and fresh apples
vere fed 3 tines a week. Water was available ad Libicun.
the {nneDrursiunrgfacteheofcontdheititohnigihnganpderi{ondtratphaelpmeobnrkaelys twuebreercutlaitntooteedstson
were conducted. Prior to nitiation of compound administration
complete physical examinations were conducted Only monkeys 1a good health were selected.
by
a
staff
veterinarian,
cerainateTdhison sJtaunduyarwyas 30i,nit1i9a78tedbecoanusJeanuofarythe10,dea1t9h7s8. of Thtehe smtoundkyeyvsa.s
2. Compound Adointstration:
Lato
5
At the end groups on a
of the random
conditioning period the monkeys were divided basis, so that the initial average body
weights were siatlar, as shown below:
Group
Nusber of Monkeys
Yale Female
Dosage Level
ng/ig/day
fo1d m
22
22
2
2
Cont1r0ol 30
iv
22
22
310000
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The test compound, suspended fn distilled water, was adainistered 7 days each veek by gavage. All doses were contained in the same voluse of water. Also the same volume of distilled water was ven to the control group. Individual daily doses were based upon the body weights, obtained weekly.
3. Observations: The monkeys were observed twice dally for general physical
appearance, behavior and pharmacotoxic signs. Individual body weights vers recorded weekly. General physical examinations were con ducted only fn the control period, because of the early deaths of all treated monkeys.
4. Clinical Laboratory Tests: Blood and urine samples were obtained for analysis from all
monkeys only in the control period. The monkeys were fasted overnight prior to the collection of blood and urine samples.
a. Hematology: Hematological studtes included: hemoglobin!, hematocrit,?
erythrocyte count3, totald and differential leucocyte counts, reticulo-
cyte count', platelet count3, prothrombin time6, activated partial
thromboplastin time' (APTT). Mean corpuscular hemoglobin, mean cor-
puscular volue and mean corpuscular hemoglobin concentration were
calculated.
b. Biochemtsery:
:
Biochentcal studies tacluded: blood glucoseS, blood urea
nitrogend, serum alkaline phosphatase, serun glutamic oxalacetic trans
aninase, serua glutamic pyruvic transaminase, cholesterol, total
protetn?, albuaind, sodiuml, potassiunl, chlorided, inorganic
phosphate, Y-glutaayl transpeptidasell, creatinine phosphokinase!2.
ha
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c. Urinalysts:
cific
Urinalysis included: measurement of volune, phl3 and spegravity; description of color and appearance; qualitative tests
for proteinl?, glucose!d, ketones!3, occult blood! and microscopic
examination of the sediment.
4. Statistical Analysis:
Yo statistical analysis on the laboratory tests was pec
formed becausa the laboratory tests were obtained for the control
period only, hovever statistical analysis of body weights were con-
ducted.
B. RESULTS:
1. General Behavior, Appearance and Survival:
The monkeys fron the 300-sg/kg/day dosage level group died
betueen the 2nd and 4th day of the study. From the lst or 2nd day the
following signs of toxicity were observed: anorexia, decreased acti-
vity (slight to severe), emesis (frothy or food like emesis) and occa
sional diarrhea. Antemortem observations were body stiffening and
stiffened limbs, general body trembling and twitching, convulsions and
prostration.
The monkeys at the 100-ng/ke/day dosage level died between the
3d and the Sth day of the study. The same toxic sysptoms as the pre-
vious group were noted fro the 2nd day.
The aduinistration of 30 ng/kg/day led to the same indications
of toxicity, but appeared some days later. The deaths also occurred
later; from the 7th to the 10th day of the study. The monkeys at the
10-0g/kg/day dosage level died between day 11 and 20 of the study.
The signs of toxicity vhich appeared in the second week of the study
were: anorexia, diarrhea, decreased activity, emesis, marked
sm,
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weakness, prostration, general body trembling and tremoring and stiffenlng of the limbs. One of the monkeys had black stools and one had erythema of the face.
There were no deaths nor unusual behavior in the control group. Occasionally, soft stools were noted.
2. Body Weights (Table 1): The monkeys from all treated groups lost body weight when the
terninal body weight vas compared with the initial body weight. In the first week of the study, there vas no statistically
significant decrease in the body weight of monkeys at the 10- and 30mg/kg/day dosage levels.
3. Laboratory Tests: 2. Hematology: The control period values are contained in Table 2. b. Blochentsery: The control period values are contained in Table 3. c. Urtnalysts: The control period values are contained in Table 4.
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Page 8 IV. PATHOLOGICAL STUDIES
A. ETHODS:
1. Gross pathology:
All experimental monkeys died during the study period. At
necropsy the heart, liver, adrenals, spleen, pituitary, kidneys,
testes/ovaries and brain vere veighed and representative tissues were
collected in buffered neutral 10% formalin. Eyes were fixed in
Russell's fixative. The thyroid/parathyroid was weighed after fixa-
tion. A mew study vas initiated (137-092) utilizing the control
monkeys from this study.
2. Histopathology:
Microscopic examination of the following formalin fixed hema-
toxylin and eosin stained paraffin sections was performed for all
animals in the experimental groups:
adrenals aorta
kLiidvneerys
pliutmubiartarsypinal cord
bbroanien
skTuingn,
stteosmtaecsh/ovaries
eeyseosphagus
mreesternotpehraircyngleyamlphlnyomdpeh ptahryartohiydroid
ghaelalrbtla(dwdietrh coronary manwodmeary gland
tthryamcuhsea
duvoedsesnevlas)
snpelreveen (with muscle) ttoonnsgiule
iJleejuumnum
pparnocsrteaatse/uterus
vuargiinnaary bladder
cocelcounn
ribmarjruonwc)tion (bone tattoo
rectun
salivary gland
and any other tissue(s) with lesions
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International Research and Development Corporation
Page 9 B. RESULTS:
1. Gross pathology (Table 5) and Organ Weights (Table 6): ALL deaths which occurred during the course of study were con-
sidered due to compound effect. Significant compound related gross pathologic findings were limited to the liver where vellowish-brown discoloration was observed in several monkeys from the 100 and 300 mg/kg/day groups. Other gross findings in these monkeys vere of agonal nature or were those which are commonly seen in untreated monkeys. While contemporary control organ weight values vere mot available for comparison, the organ weight values of these monkeys appeared to be within normal limits.
2. Histopathology (Table 7): No consistent histopathologic changes which vere considered
to represent a direct compound effect were observed in any tissues examined microscopically. Many agonal changes, including congestion, hemorrhage and lipid depletion of the adrenal cortex, vere seen in monkeys from all experimental groups. No microscopic basis was seen for the yellowish-brown discoloration noted at necropsy in several monkeys from the 100 and 300 mg/kg/day groups.
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FFlluuoorroacdheemical
_
Paaggee 10
_Sur--factant TC-95: 9-Day Subacute Rhesus Yomkey Toxicity Study.
TABLE 1.
Individual Body Weightese, Kilograms.
Group,
M--Nounmk-- beeeryr------s--ex------T--Co--ntr--ol.--7 --------T ------W2--eek--of--St--udy--3 --------
Control:
7355 7358
u u
7337628
FF
Mean
10 ng/kg/day:
73%7 7356
u u
7369
7371
Mean
30 mg/kg/day:
739 7351
u x
7370 7377
F
Mean
100 mg/kg/day:
3.10 3.7
3.10 7
3.10 37
3.20 37s
33..5055 33..1600 33.6100 33.6255
3.35 3.39 3.38 3.46
3.40 3.05
3.40 310
3.05 3.05
2.90 2.65
2.50% Died 2.40% Died
3.10 3.10 3.85 4.00
2.05 370
2.70 3.25%
Died
2.55%
Dies
3.35 3.40 3.21 2.75
33..6455 33.475 33.0055
3.05 3.50
3.20 3%
2.60 2.95
341 3.39 2.91
23..9100%% DDiieedd 2.65% Died 2.50% Died
73735596
Mu
7373 7374
F F
Mean
300 mg/kg/day:
33.800 33.2800 23..8805 32..8800 3.39 3.38
23..7500%% DDiieedd 23..4250%% DDiieedd
77336507
uu
33..0600 3.30655 23..8400%% DDiieedd
7375 7379
F F
4.05 2.70
"10 2.75
3.35% Died 2.60% Died
Mean
3% 3.39
137-087
Terminal weight not included in mean.
002082,
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50-ay Subacute Rhesus Monkey Toxkeicy Sealy.
Microscopic Observations, Deaths.
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002092
Page 21 BpT s rEeeeRe re e ETesev Tn e EmE wCoE m .--m --enier Te m me none
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