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Toxicological information

Skin irritation / corrosion

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Administrative data

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Jun - Nov 2001
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
The objective of the study is to obtain information on the local eye irritation and the systemic toxicity of Diovan eye drops (5%) when administered two or four times daily for 14 days into the conjunctiva! sac of albino rabbits. Four groups of 4 male and 4 female rabbits each were used.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2001
Report date:
2001

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
The study was performed in compliance with:
'Good Laboratory Practice' Regulations of the EC enacted in Germany in the 'Chemikaliengesetz' [Chemicals Act], current edition;
United States Food and Drug Administration Good Laboratory Practice Regulations, 21 Code of Federal Regulations, Part 58, current edition.
The following regulations were considered:
'OECD Principles of Good Laboratory Practice' (as revised in 1997) ENV/MC/CHEM (98) 17, January 21st, 1998;
Japanese Guidelines for Non-clinical Studies of Drugs Manual 1995; Guidelines for Toxicity Studies of Drugs. Japanese Ministry of Health and Welfare.
GLP compliance:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
N-Pentanoyl-N-{[2'-(1H-tetrazol-5-yl)biphenyl-4-yl]methyl}-L-valine
EC Number:
604-045-2
Cas Number:
137862-53-4
Molecular formula:
C24 H29 N5 O3
IUPAC Name:
N-Pentanoyl-N-{[2'-(1H-tetrazol-5-yl)biphenyl-4-yl]methyl}-L-valine
Test material form:
solid: particulate/powder

Test animals

Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
32 purebred New Zealand white rabbits ( 1 6 males and 1 6 females) were used for this experiment. Before start of study a thorough examination was given to a pool of 36 animals. Animals were judged to be healthy on the basis of general observations (physical/auditory, ophthalmoscopic- and laboratory examinations). Thereof 32 animals with parameters in the normal range were inserted in this study.
Diet
ALTROMIN 2023 (supplier: ALTROMIN GmbH, D-32791 Lage/Lippe; see Appendix 3:
Composition of the diet) served as food. This food was offered ad libitum. Food
residue was removed and weighed.
Samples of the food are analysed for contaminants based on EPA/USA 1 by AUA2
twice a year (see Appendix 3: Limitation for contaminants in the diet). Certificates
of analysis of the composition and for contaminants were provided by the manufacturer
and are included in the raw data.
Drinking water
Tap water of drinking water quality was offered ad libitum.
Samples of drinking water are taken twice a year by the Wasserbeschaffungsverband
Harburg, D-21220 Seevetal, and are analysed according to the 'Deutsche Trinkwasserverordnung,
Bundesgesetzblatt, Jahrgang 1990' [German Regulations on
drinking water, public notice of the law, 1990] (see Appendix 3: Limitation for
contaminants in the drinking water).
In addition, drinking water samples taken at LPT are analysed by LUFA-ITL3 once a
year for means of bacteriological investigations according to the 'Deutsche Trinkwasserverordnung,
Bundesgesetzblatt, Jahrgang 1990, Anlage 4'

Test system

Type of coverage:
other: instillation into the conjunctiva! sac of the left and right eyes
Preparation of test site:
other: Then the lower lids of the eyes were carefully pulled away from the eyeball and the test or control substance was instilled into the conjunctiva! sac.
Vehicle:
other: Diovan eye drops vehicle
Controls:
yes, concurrent vehicle
yes, concurrent negative control
Amount / concentration applied:
Group 1 (negative control)
- right eye: 0.9% NaCl solution
- left eye: 0.9% NaCl solution
1 drop each eye,
4 times a day for 2 weeks, at 2-hours intervals
Group 2 (vehicle control)
- right eye: vehicle
- left eye: vehicle
1 drop each eye,
4 times a day for 2 weeks, at 2-hours intervals
Group 3
- right eye:
- left eye:
Diovan eye drops (5%)
Diovan eye drops (5%)
1 drop each eye,
2 times a day for 2 weeks, at a 6-hours interval
Group 4
- right eye:
- left eye:
Diovan eye drops (5%)
Diovan eye drops (5%)
1 drop each eye,
4 times a day for 2 weeks, at 2-hours
intervals
1 drop ~ 25 μI/instillation/eye
The test formulation was very viscous and hardly droppable.
For instillation of the eye drops the following pipettes were used:
CP 100 ( 10-100 μI) for Mic roman M 100 (GILSON)
Duration of treatment / exposure:
Treatment for 2 weeks
1 drop is cca 25ul
Observation period:
3 adaptation weeks
14 test days
Number of animals:
32 purebred New Zealand white rabbits ( 1 6 males and 1 6 females) were used for
this experiment. Before start of study a thorough examination was given to a pool
of 36 animals. Animals were judged to be healthy on the basis of general observations
(physical/auditory, ophthalmoscopic- and laboratory examinations). Thereof 32
animals with parameters in the normal range were inserted in this study.
Details on study design:
The animals were allocated to the 4 groups by randomisation:
Treatment for 2 weeks
right eye left eye No and sex of animals
____________________________________________________________________________________
0.9% NaCl solution 0.9% NaCl solution 4M, 4F
1 drop per instillation, 1 drop per instillation,
4 times/day, at 2-h intervals 4 times/day, at 2-h intervals
_____________________________________________________________________________________
Vehicle Vehicle 4M, 4F
1 drop per instillation, 1 drop per instillation,
4 times/day, at 2-h intervals 4 times/day, at 2-h intervals
____________________________________________________________________________________
Diovan eye drops (5%) Diovan eye drops (5%) 4M, 4F
1 drop per instillation, 1 drop per instillation,
2 times/day, at a 6-h interval 2 times/day, at a 6-h interval
____________________________________________________________________________________
Diovan eye drops (5%) Diovan eye drops (5%) 4M, 4F
1 drop per instillation, 1 drop per instillation,
4 times/day, at 2-h intervals 4 times/day, at 2-h intervals

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Remarks on result:
not measured/tested
Irritation parameter:
overall irritation score
Basis:
animal #1
Time point:
24/48/72 h
Remarks on result:
not measured/tested
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Remarks on result:
not measured/tested

Any other information on results incl. tables

Ophthalmological examination

No test substance-related changes of the eye and the optic region were observed for any of the tested groups.

The animals did not show any changes to conjunctivae, cornea, anterior chamber, iris, lens or fundus. Miosis, mydriasis or ptosis were not observed. The fluorescein test revealed no test substance-related changes in any of the rabbits.

Ophthalmopathology

The histopathological examination did not reveal any test substance-related morphological changes of the eyes, eyelids and eye-adnexa of all animals examined. There were no differences between the test substance-treated and the control animals.

Systemic tolerance

Behaviour, external appearance, faeces, mortality

Diovan eye drops vehicle administered 4 times/day or Diovan eye drops (5%) administered 2 or 4 times/day did not influence the behaviour and external appearance in any of the animals. None of the rabbits died prematurely.

Body weight

No test substance-related influence was observed on the body weight for any of the tested groups.

Food and drinking water consumption

No test substance-related influence was observed on food consumption for any of the tested groups.

The visual appraisal of the drinking water consumption revealed no differences between the negative control and the animals treated with Diovan eye drops vehicle 4 times/day or Diovan eye drops (5%) 2 or 4 times/day.

Haematology

Treatment with Diovan eye drops vehicle 4 times/day or Diovan eye drops (5%) 2 or 4 times/day did not cause any test substance-related changes to the haematological parameters.

No influence was observed for the differential blood count, the haematocrit value, haemoglobin content, number of erythrocytes, leucocytes, reticulocytes and platelets, the thromboplastin and activated partial thromboplastin time, mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC).

Clinical biochemistry

Treatment with Diovan eye drops vehicle 4 times/day or Diovan eye drops (5%) 2 or 4 times/day did not cause any test substance-related changes to the clinicobiochemical parameters.

No influence was observed on the plasma or serum levels of bilirubin (total), cholesterol (total), creatinine, glucose, phosphate (inorganic), protein (total), triglycerides, urea (in blood), calcium, chloride, potassium and sodium.

All results were within the normal limits of the negative control. The serum protein electrophoresis revealed no test substance-related changes. The activity of the alanine aminotransferase (ALAT /GPT), the aspartate aminotransferase

(ASAT/GOT), the alkaline phosphatase (aP) and the lactate dehydrogenase (LDH) was not increased.

All relative organ weights were compared statistically. No test substance-related differences were observed from the negative control for animals treated with Diovan eye drops vehicle 4 times/day or Diovan eye drops (5%) 2 or 4 times/day.

Histopathology

The histopathological examination did not reveal any test substance-related morphological changes in any ot the organs examined.

Bone marrow evaluation

No test substance-related changes were noted on the myeloid : erythroid ratio.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Based on the findings of the study, repeated 2-week instillation of Diovan eye drops (5%) two or four times daily into the left and right eye of albino rabbits resulted in no local or systemic intolerance reactions.