11β,16α,17,21-tetrahydroxypregna-1,4-diene-3,20-dione

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

2010

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP-compliant guideline study, available as unpublished report, no restrictions, fully adequate for assessment. The restriction is due to the use of the read across approach: the test was performed not with hydroxyprednisolone but with prednisolone, a substance which is very similar in structure and physical/chemical properties.

Data source

Reference

Reference 1

Reference Type:

other: Protocol Draft

Title:

Draft Updated ICCVAM Recommended HET-CAM Protocol

Author:

ICCVAM

Year:

2010

Bibliographic source:

ICCVAM Test Method Evaluation Report: Current Validation Status of In Vitro Test Methods Proposed for Identifying Eye Injury Hazard Potential of Chemicals and Products - NIH Publication No. 10-7513

Report date:

2010

Reference 2

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Version / remarks:

ANNEX - A Sequential Testing Strategy for Eye Irritation and Corrosion .

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Draft Updated ICCVAM Recommended HET-CAM Protocol.

Principles of method if other than guideline:

Results from in vitro or ex vivo tests: Substances that have demonstrated corrosive or severe irritant properties in an in vitro or ex vivo test that has been validated and accepted for the assessment specifically of eye or skin corrosivity/irritation, need not be tested in animals. It can be presumed that such substances will produce similar severe effects in vivo.
Even if HET-CAM assay heve not achieved the OECD validation, it is accepted for the purpose of "TESTING AND EVALUATION STRATEGY FOR EYE IRRITATION/CORROSION" as reported in ANNEX of test method B.5.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Prednisolone

EC Number:

200-021-7

EC Name:

Prednisolone

Cas Number:

50-24-8

Molecular formula:

C21H28O5

IUPAC Name:

11,17,21-trihydroxypregna-1,4-diene-3,20-dione

Test material form:

solid: particulate/powder

Details on test material:

- Storage conditions of the test material: at room temperature, protected from light

Test system

Vehicle:

physiological saline

Remarks:

B. Braun Melsungen, lot n. 017A122, expiry date: 03/2013.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Prednisolone 100 % applied to an area of 25 % of the chorioallantoic membrane.

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.3 mL of standard solution (0.9 % NaCl)
- Lot/batch no. 017A122

Duration of treatment / exposure:

The reaction on the CAM were observed over a period of 5 minuted monitoring the appearance of:
Haemorrhage (bleeding);
Vascular lysis (blood vessel disintegration);
Coagulation (protein denaturation intra- and extravascular).

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done):
- Time after start of exposure: Prednisolone was left in contact with the egg membrane for 5 minutes.

SCORING SYSTEM: the time for each reaction to occur was recorded and an irritation score (IS) was calculated:
IS = [(301-sec H)/300] x 5 x S+ [(301-sec L)/300] x 7 x S + [(301-sec C)/300] x 9 x S

sec = second of first occurence of reaction
H = Haemorrhage
L = Vascular Lysis
C = Coagulation
S = 0.1 if H, C, L is grade 1 (weak reaction)
S = 0.5 if H, C, L is grade 2 (moderate reaction)
S = 1 if H, C, L is grade 3 (strong reaction)
The mean score was calculated from irritation scores for each egg for each test group.

TOOL USED TO ASSESS SCORE: hand-slit lamp / biomicroscope / fluorescein

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean

Value:

0

Remarks on result:

no indication of irritation

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The eye irritancy potential of the test item was investigated in the chorioallantoic membrane assay.
The test item was tested as provided by the sponsor in order to find any irritancy potential.
The calculated mean irritation score was 0.
The test item was classified as non-irritant.
The positive and negative controls were within the historical control data range demonstrating the validity and sensitivity of the test.

Executive summary:

This in vitro eye irritation test was performed with prednisolone, a compound which has been demonstrated to be very similar in structure, physicochemical properties and toxicological profile to hydroxyprednisolone in the Analogue Approach - Read across Hydroxyprednisolone document (2013, see section 13). Due to the fact hydroxyprednisolone and prednisolone have nearly the same chemical structure, the same interaction with bio-molecules, living cells and tissue and metabolic pathway is expected. Therefore, a read across from hydroxyprednisolone to data obtained with prednisolone is scientifically justified.