2.2 - Treatment

PHYCHER Bio dhdoppement
Study: TA0423-PH-03/030?
pcln
2.2 - Treatment
The animals of Group 2, received an effective dose of 5000 mgkg body weight of product,
administered by force-feeding under a volume of Sn-Ukg body weight using a suitable syringe
graduatedfitted with an oesophagealmetal canula.
3-RESULTS
No mortality occurredduring the study.
The clinical observationsare shown in the observationsdata sheet, tables 1 to 2 and those of body
weight evolution in tables 3 and 4, hereafter.
No clinical signs related to the administration of the test product were observed.The body weight
evolution of the animals remainednormal throughout the study, similar betweentreated and control
animals.
The results of the macroscopical examinationsare shown in the necropv data sheet, tables 5 to 8,
hereafrer.
The macroscopicalexaminationof the animals at the end of the study did not reveal treatment-related
changes.
4-CONCLUSION
The LDSo of the product REF.03.749CENNAiWDE CERO 15 is higher than 5000 mgkg body
weight by oral route in the rat.
According to the criteria for classl?cation, packaging and labelling of dangeroussubstances and
preparations in accordance with the iL?.E.C.Directives 67/548, 2001/59 and 991’45,the product
REF.03.749CENNAMIDE CERO15 must notbe classified.
S-ARCHIVES
All original data and the final report related to this study will be stored for a period of 10 years in the
company under the referenceTAO423-PH-03/0307.After this period, the Sponsor’sinstructions will
be applied.
6-PROTOCOLADEtERENCE
Environmental parameter:A temperatureof 18°C was registered during this study instead of 19°C
(minimal lim it) as plannedin the experimentalprotocol.
This deviation did not, in any case,influence the developmentand the resultsof the study.
Study: TAO423-PH-0310307
PA
PHYCHER Bio dLveIoppemen!
* SpontaneousActivity :
* Preyer’s Reflex :
*Respiratory Rate :
* Convulsions :
* Tremors :
* Body Temperature:
* Muscle Tone :
Normal @ I) 1 Decrease(D) 1 Increased(A)
Normal (N) 1 Decrease@) 1-Increased(A) 1 None (0)
Normal (N) 1 Dyspnea(D) 1 Bradypnea(B) 1 Polypnea(P)
None CN) 1 Tonic (T) 1 Clonic (C)
None (N) I Tremors (Tr)
Normal (N) I Hypothermia(D) 1 Hyperthermia (A)
Normal CN) 1 Decreased(D) I Increased{A)
* PalpebralOpening :
Normal (N)
Eyes partly closed (PC)
Eyes completely closed (Cc)
* Pupil Appearance:
Normal @J) I Mydriasis (Md) 1 Myosis (MS)
* Salivation :
* Lachrymation :
* Righting Reflex :
* Back Hair Appearance:
NOTES :
l
l
Normal (N) 1 Increased(A)
Normal (N)
Normal (N)
I Increased(A)
I Limited @) I None (0)
Normal (N) 1 Piloerection (Pi)
The observationsare perjormed by comparisonwith the control
animals standing in the sameenvironment.
The temperatureof animal is evaluatedby touch.
7116
PHYCHER Bio dJveloppement
Study: TAO423-PH-03/0307
&g
TEST PRODUCT: REF.03.749 CENNAMIDE CERO 15
1.:
Treatment:5000 mg&g body weight
Op&ators: F. Richeux
G. Creste
DO: 10/07/03(females)
lo/lo/O3 (males)
Number of animals: 6
Table 1
OBSERVATIONS :
?.
kespiratoryrate
Ionvulsions
liemors
Body temperature
Muscle tone
Palpebralopening
Pupil appearance
Salivation
Lachrymation
Righting reflex
Remarks
None
None
PHYCHER Rio d&eloppement
TEST PRODUCT:
L
Study: TAD423-/‘H-03/0307
P&
REF.03.749 CENNAMIDE
CERO 15
Treatment:5000 mg/kg body weight
OpCrators:F. Richeux
G. Creste& A. Ehanno
DO: 10/07/03(females)
1O/1O/O3(males)
Number of animals: 6
:
Table 2
OBSERVATIONS :
iespiratory rate
Zonvulsions
rremors
Body temperature
Muscle tone
Palpebralopening
Pupil appearance
Salivation
Lachrymation
Righting reflex
PHYCYiEB Bio dbveloppement
Study: TA0423-PH-03/0307
,p,lalfl
CONTROL PRODUCT : DISTILLED WATER
*-I
Treatment: 10000mgkg body weight
Operatars:G. Creste
A. Ehanno
DO: 1O/07/03(females)
10/10/03(males)
Number of animals: 6
Table 3
(Body weight and weight gain in grams)
MALES
Rm 4049
Rm 4050
Rm 4051
I’
MEAN
f-
Standard deviation
FEMALES
Rf 4031
Rf 4032
Rf 4033
MEAN
Standard deviation
PHYCffER Bio dt?veloppement
Study: T&0423-PH.03/0307
&i-g
TEST PRODUCT: REF.03.749CENNAMIDE CERO 15
:.
Treatment:5000 mg/kg body weight
Operators:G. Creste
S. Uteau & A. Ehanno
DO: 1O/07/03(females)
lo/lo/O3 (males)
Number of animals: 6
Table 4
(Body weight and weight gain in grams)
Standard deviation
PHYCHER Bio dheloppement
Study: TAO423-P&03/0307
pottl
CONTROL PRODUCT : DISTILLED WATER
Dose: 10000 mgikg body weight
s ?:
Operator: A. Ehanno
Route: oral
Sex / No. animals: 3 males Rm4049 to I&405 1
Necropsy date: 1O/24/2003
Species: Sprague Dawley Rats
Tuble 5
pontaneousdeath :
1-1
;ENERAL APPEARANCE
Euthanasia :
BEFORE AUTOPSY
[xl
At term
ixf
: No-1
Remarks
@ESOPHAGUS
’STOMACX
’DUODENUM
’JEJUNUM
’ILEON
’CAECUM
i COLON
t RECTUM
?ARTICULARS
: None
PHYCHER Bio dheloppement
Study: TAO423-PH.03/0307
m
CONTROL PRODUCT : DISTILLED WATER
Dose: 10000 mgkg
body weight
Operator: G. Creste
Route: oral
Necropsy
Sex / No. animals: 3 females Rf403 1 to Rf4033
date: 1012 l/2003
Species: Sprague Dawley
Table 6
Spontaneous death :
1-1
GENERAL APPEARANCE
Euthanasia :
BEFORE AUTOPSY
X
X
X
X
X
X
X
X
X
X
X
* LXVER
*THYMUS
* TRACHEA
* LUNGS
* HEART
*
*
*
*
*
KIDNEYS
URINARY BLADDER
TESTICLES
OVARIES
UTERUS
* TREATMENT
* ADRENALS
* PANCREAS
PARTICULARS
X
X
X
X
X
X
X
AREA
X
X
: None
.V.t. R. : Nothing to report
At term
[Ti
: Normal
Observed
Organs
’ (ESOPHAGUS
* STOMACH
@DUODENUM
c JEJUMJM
L ILEON
lcCAECUM
L COLON
‘RECTUM
@SPLEEN
[x-1
Results
Remarks
Rats
PHYCHER Bio d&foppentent
Study: TAO433-PH-03/0307
paril
TESTPRODUCT:REF.03.749CEN-NAMIDECERO15
Dose: 5000 mg/kg body weight
Operator: A. Ehanno
Route: oral
Sex / No. animals: 3 maies Rm4064 to Rm4066
Necropsy date: 10/24/2003
Species: Sprague Dawley Rats
Table 7
Spontaneousdeath :
L-1
Euthanasia:
171
At term
[T]
GENERAL APPEARANCE BEFORE AUTOPSY : Normal
Observed
Organs
Results
* CESOPHAGUS
* STOMACH
* DUODENUM
*JEJuNuM
* ILEON
* CAECUM
* COLON
*REcMJM
* SPLEEN
* LIVER
“TIIYMUS
X
X
X
X
X
X
X
X
X
X
X
N.t.R
N.t.R
N.t.R
*TRACHEA
* LUNGS
* HEART
X
X
X
X
N.t.R.
N.t.R
N.t.R.
*
*
*
*
*
KIDNEYS
URINARY BLADDER
TESTICLES
OVARIES
UTERUS
X
X
* TREATMENT AREA
* ADRENALS
* PANCREAS
PARTICULARS : None
N.t.R. : Nothing to report
X
X
NJ-R.
NsR.
N.t.R
NAR.
N.t.R.
N.tR
N.t.R
Nl.R
N:tR
N.t.R.
N.t.R.
Remarks
PHYCHER Bio ddveioppentent
Study: TAU$23-PH-03/U30?
paril
TESTPRODUCT:REF.03.749CENNAMJDECEROl5
Dose: 5000 mg/kg body weight
Operators: G. Creste
Route: oral
Sex I No. animals: 3 females Rf4046 to lW4048
Necropsy date: 1O/2 112003
Species: Sprague Dawley Rats
Table 8
s pontaneousdeath :
f-1
Euthanasia:
[XI
At term
GiENERAL APPEARANCE BEFORE AUTOPSY : Normal
Observed
organs
:
r
CESOPHAGUS
STOMACH
DUODENUM
JEJUNUM
ILEON
CAECUM
COLON
‘RECTUM
’SPLEEN
’LIVER
‘THYMUS
X
X
X
X
X
X
X
X
X
X
X
N.tR.
N.f.R
N.t.R.
N.t.R.
N.tR
N.tR.
N.tR
N.t.R.
N.tR
N.t.R.
N.f.R
’TRACHEA
i LUNGS
FHEART
X
X
X
X
N.t.R.
N.t.R.
N.t.R.
k KIDNZYS
k URINARY BLADDER
k TESTICLES
c OVARIES
k UTERUS
X
N.t.R.
N.t.R.
X
X
N.tR.
N.t.R.
x
X
N.t.R.
* TREATMEANT AREA
* ADRENALS
* PANCREAS
PARTICULARS : None
N.t.R. Nothing to report
NtR
1x1
PHYCHER Bio dheioppement
Study: T&0423-PH-03/0307
m
Appendix 1: Statement of GLP Compliance (1 page)
Appendix 2: Animals diet sheet (1 page r/v)
TM
:01534+
968210
OBJET : Evaluation de la conformit aux B.P.L. selon la directive 88/320/CE.E.
Cons&xtivement B votre engagement vis & vis du GIPC et du COFRAC et en applicationdu d&ret
no 81-278 du 25 mars 1981 portant c&ion d’un groupe intenninisthiel des prod&s chimiques,
modifi6 nhmment par Ie d&ret 90-206du 7 mars 1990et par le d&ret no 98-l 3 12 du 3 1 dkembre 1998
concernantles bonnes pratiquesde laboratoires, je vous confirtne que le GIPC, au vu des rC.sultatsdu
contrdle exercCpar le Comitk fran@s d’acctiditation (COFRAC) - Section Essaisa dkidd pour votre
installation du statut s&ant :
respect des principes de B.P.L.
Domaines de reconnaissance :
:
,G;
a 4-
I: jCl 6il 7a sCl9
essaisphysico-chimiques
&tides de toxicit
&udes de mutagknicit4
etudes kcotoxicoiogiques SW les organismes aquatiques et terrestres
&da portant sur le comportement dans I’esu, dans le sol et dans l’air ; bioaccumulation
ttudes portant SW les kidus
ttudes portant sur les effets sur les mksocosmes et les tcosystkmes naturek
m&odes de chimie anatytique et clinique
autres Ctudes
Date d’inspection : 19 - 20 mai 2003
a inspection
initiale (i.i)
Ll inspection de contr6le piriodique (i.p)
0 inspection de contr6le compknentaire (kc)
!TJinspection d’extension (i.e)
.&inspection de renouveilement (Lr)
Date de dkision du G.I.P.C.
Date de prise d’effet :
:
25 mai 2003
20 mai 2003,
hnde de premihe conformit : 1998
28‘novembre2004
Date de validit :
310 DEVELOPPEMENT
Lou Tribail
2.1. de Toctoucau
PHYCHER
18, chemin
33611 CESTAS Cedex
: ,404~70
1Pellets 0 15 mm
‘%dSarU tmk
Rate per day
rat 15to25g.
.--__ ._ .-J.
Formula
Formule
.. ..*.-a~*W‘JVlGW
.ru,uE~~PYg,swllSoe3P
tug.
Average analysis
Anaiyse
Amino acid values Icalculated/kg)
Apport
,
en acides amin&
(CUICUI~/~~)
moyenne
Fatty acid values icahJated/kg)
&port
en acides gras (dctd&/kg)
Mineral and vitamin content
Con)enu
P
CO
Na
K
mg
mg
mg
mg
m!3
mg
43
Mb7
Fe
CU
zn
“9
mg
f-3
co
I
-3
mg
5900
-
3300
300
6700
5ooo
1600
-
ISa,
100
50
90
15
A0
150
15
45
40
Traces
0‘3
1,5
-
mint&at
foTAL
5900
8300
1900
6700
zoo0
90
240
30
85
1.5
0,3
et vifaminique
scakrrl ad/kg
KG m/kg
Gx
tMV vat
Warn.
Warn.
Vitam.
Warn.
Warn.
Warn.
Warn.
Warn.
A
D3
Bl
52
83
56
512
E
Vitom. K3
Viium. PP
AC. Folic
Biotine
Choline
UI
UI
mg
T3
“9
2
-3
ms3
*g
mg
*g
-3
-.
Traces
Traces
7500
2
10
13
0,Ol
4,5
6,5
I,3
0,Ol
15
2,25
15
0
0
400
15
0,25
60
0,s
0,Od
1 200
1500
1
1
TOTAL
7500
1500
7
6,5
16,5
76
0,02
30
23
75
OS
0,04
1 600
i
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i
PHYCHER Bio dheloppentent
Study:TAO423-PH-03/030?
w
I
CONTENTS
f
PART 1
QUALITY ASSURANCE ATTESTATION
Page 3
AUTHENMCATION
Page 4
SUMMARY AND CONCLUSION OF THE STUDY
Page 4
TEST REPORT
Page 5
1 - TEST PRODUCT
Page 5
2 - STUDY DEVELOPMENT
Page 5
2.1 - Animals
2.2 - Treatment
Page5
Page6
3 - RESULTS
Page 6
4 - CONCLUSION
Page 6
S - ARCHIVES
Page 6
6 - PROTOCOL ADHERENCE
Page 6
TABLES OF RESULTS
Pages8- 15
PART 2
APPENDICES
Appendix 1: Statementof GLP Compliance (1 page)
Appendix 2 : Animals diet sheet (1 page)
Page 16
I
i
2: IG
PHYCHER
Bio dheloppement
OUALITY
Szudy: TA0423-PM03/0307
parrl
ASSURANCE ATTESTATION
I, the undersignedBernard Benech, Quality Assuranceunit of PHYCHER Bio dkveZoppement, attest
that the study TAO423-PH-03/0307was submitted to the inspectionof Quality Assurance.
The routine inspections of the toxicity tests performed within the laboratory of PHYCHER Bio
dbeloppement, are carried out as a control processof different main technical phasesconcerning at
least one similar test. The frequency is once a term or more. The date(s) of technical phases
inspectionsrelating to this test are given below:
Date (s) of inspection (s)
: October, 17” ,2003
Clinical examinations at 48 hours (males)
Date of reporting to Study Director
Date of reporting to Management
: October, 20’, 2003
: October,20’, 2003
This report has been audited by PHYCHER Bio dkvefoppement Quality Assurance unit. It is
considered to be an accurate account of the data generatedand the application of the operating
proceduresin use within the laboratory.
Date (s) of inspection (s)
Date of reporting to Study Director
Date of reporting to Management
Date : December 2”d, 2003
Final report
: December,2”, 2003
: December,2&, 2003
: December,2*, 2003
PHYCHER Bio dheloppement
Study: TAO423-PH-0310307
&&
AUTHENTICAlXON
I, the undersigned,FrancoisRicheux, Study director, certify that the study TA0423-PH-03/0307was
performedon the premisesof the LaboratoryPHXHER Bio developpement.
*.
I, certify that the objectives laid down in the technical protocol were achievedand no undesirable
event occurred to affect the quality or the integtity of the study. I consider the data generatedto be
valid. This report fully and accuratelyreflects the operatingproceduresusedin the laboratory and data
generated.
The entire work was devisedand performedin compliancewith the principles of the Good Laboratory
Practice (G.L.P.), as defined in the O.E.C.D. ruling reiative to the mutual acceptanceof data in the
evaluation of chemical substances (C(81) 30 @ tat’) Appendix 2 - May 12”, 1981; C (97) 186,
November 2@, 1997), and transcribed in the decree n* 98-1312 dated December31’! 1998 of the
Journal Oflciel de la RepubliqueFrancaise.
Date : December20d,2003
I
SUMMARY
AND CONCLUSION
OF THE STUDY
The product REF.03.749 CENNAMIDE CERQ 15 was administered to a group of 6 Sprague
Dawley rats (3 males and 3 females) at the single dose of 5000 mg/kg body weight according to the
experimental protocol establishedon the basis of the official method as defmed in the O.E.C.V.
guideline W 423 dated March 2Td. 1996 and the test method B.lter of the Directive No 96/541!EC
dated JuIy 3@, 1996.
.
No mortality occurredduring the study.
No clinical signs related to the administration of the test product were observed.The body weight
evolution of the animals remainednormal throughoutthe study, similar betweentreated and control
animals.
The macroscopicalexaminationof the animals at the end of the study did not reveal treatment-related
changes.
In conclusion, the L&J of the product REF.03.749CENNAMDE CERO 15 is higher than 5000
mg/kg body weight by oral route in the rat.
According to the criteria for classification, packaging and labelling of dangeroussubstances and
preparations in accordance with the E.E.C. Directives 67I.548,2001/59 and 99/45, the product
REF.03.749 CENNAMIDE CERO 15 must not be classified.
Date : December2”d : 2003
ment
Pharmacologist
@W
4'16
PHYCHER Bio dheloppement
Study: TAOd23-P&f-03/030?
pclrsl
TEST REPORT
1 - TEST PRODUCT
The product REF.03.749 CEIWAMIDE CERO 15, sent by BIQ-HC - Pam d’activitb de
Canteranne- Bat 3 - Avenue de Canteranne - 33600 PESSAC, was received on 09/27/2003. Its
characteristicswere :
l
l
l
l
Container : plastic pot
Quantity : 83.32 (container+ contents)
Batch
: 030824
CASN” :-
.
*
0
.
Form : liquid
c010ur :brown
Storage : room temperature
Purity :-
It was identified under the code number: PF&O3/0307.
Information concerning the identity, purity and stability of the product to be studied are the
responsibility of the Sponsorof the study. No information concerningthe product was provided by the
sponsor.
2 - STUDY DEVELOPMENT
The study was carried out between 10/W/2003 and 10/24/2003, in PHYCk&??RBio.dkveloppement - 18
chemin Lou Tribail - Z.I. de Toctoucau - 33611 Cestascedex, accordingto the experimentalprotocol
Ref: TA0423-version December 2000, establishedaccordingto the O.E.C.D. guide&e N” 423 dated
03.22.1996 concerning acute oral toxicity and the test method B.lfer of the Directive N” 96/5d/EC
dated 07.30.1996.
The animal control group was performed between 10/07/2003and 10/21/2003according to the same
experimental conditions, with administration of the control product (Distilled water), under a volume
of 1M /kg body weight.
2.1- Animals
12 SpragueDawley rats (SPF Caw) originating from Elevage Janvier (53940 Le Genest-St-Isle France),were used after a 5 to 6-&y acclimatisationperiod. At the beginning of the study, the animals
weighted between 184g and I97g (males) and between156g and 170g(females).
Groun 1 (control1:
3 male rats
and 3 female rats
Rm4049 ii Rm4051
Rf4031 to Rf4033
Groun 2 (treated):
3 male rats
and 3 female rats
Rm4064 to Rm4066
W4046 to Rf4048
Environmentalparametersfor the treatedgroup :
- temperature :
between18°C and 22’C
- relative humidity : between40% and 67%
5’!6