Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
7 March 1989 - 4 April 1989
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was peformed according to OECD guidelines and GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1989
Report Date:
1989

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Name: M-3175; 1,3-propylenediamine-N,N,N',N'-tetra acetic acid, ammonium iron (+3) salt (mentioned in 28-day study report)
Appearance: yellow powder
Batch No.: VG-01
Purity: 99% (mentioned in 28-day study report)

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Olac Ltd., Bicester, Oxfordshire, England
- Age at study initiation: no data
- Weight at study initiation: 205 - 266 g
- Housing: The guinea-pigs were housed in stainless steel cages measuring 55 x 60 x 23 cm, with stainless steel grid floors (Modular Systems and Development Co. Ltd., London, England). The grid floors ensured rapid removal of waste material to undertrays which were cleaned out as necessary. Up to five animals of the same sex were assigned to each cage. The cages were held in mobile tubular steel racks.
- Diet (e.g. ad libitum): A commercially-available pelleted guinea-pig diet (Guinea-pig F.D.L., from special diet service limited, Witham, Essex, England) was fed without restriction.
- Water (e.g. ad libitum): Animals had free access to tap water supplied via an automatic watering system
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 15-23
- Humidity (%): 40-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: 7 March 1989 - 4 April 1989

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
First induction:
5% w/v M-3175 in distilled water
5% w/v M-3175 in FCA
Second induction:
30% w/v M-3175 in distilled water
First challenge:
10% w/v M-3175 in distilled water
3% w/v M-3175 in distilled water
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
First induction:
5% w/v M-3175 in distilled water
5% w/v M-3175 in FCA
Second induction:
30% w/v M-3175 in distilled water
First challenge:
10% w/v M-3175 in distilled water
3% w/v M-3175 in distilled water
No. of animals per dose:
10 males and 10 females
Details on study design:
RANGE FINDING TESTS:
Intradermal administration of a range of concentrations from 0.5% to 30% w/v M-3175 in distilled water or FCA caused slight or moderate erythema. Concentrations of 10% and 30% w/v M-3175 also caused eschar formation. It proved difficult to administer 30% w/v M-3175 in FCA.
In the first topical screen, occluded application of 50% w/v M-3175 in distilled water caused slight erythema, eschar formation and exfoliation. No reaction was observed following similar application of 30%, 10% or 5% w/v M-3175 in distilled water. In the second topical screen, occluded application of 30% w/v M-3175 in distilled water caused slight erythema and eschar formation in one animal. No reaction was observed following similar application of 10%, 5% or 3% w/v M-3175 in distilled water.
After consideration of the results of the irritation screens and the criteria for main study concentrations selection, the following treatment regime was chosen.
First induction:
5% w/v M-3175 in distilled water
5% w/v M-3175 in FCA
Second induction:
30% w/v M-3175 in distilled water
First challenge:
10% w/v M-3175 in distilled water
3% w/v M-3175 in distilled water

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures:
Intradermal: Three pairs of injections (0.1 ml).
Injection sites - Test group treatment - Control group treatment
Anterior sites - FCA - FCA
Middle sites - 5% w/v M-3175 in distilled water - Distilled water
Posterior sites - 5% w/v M-3175 in FCA - Distilled water in FCA

On Day 8 the dermal areas treated on Day 1 were treated by topical application of 0.6 ml 30% w/v M-3175 in distilled water in test animals while controls received 0.6 ml distilled water. Each dose was absorbed onto a 4 x 2.5 cm absorbent patch (Whatman No. 3 filter paper) which was
applied to the skin and covered by an occlusive dressing (Blenderm, 3-M Co. and Elastoplast, T. &J. Smith and Nephew Ltd) for 48 hours.

- Test groups: 10 males 10 females
- Control group: 10 males 10 females
- Site: back
- Frequency of applications: see above
- Duration: see above
- Concentrations: see above

B. CHALLENGE EXPOSURE
Both flanks of all animals were shaven to expose areas (5 x 5 cm) on either side of the trunk. On Day 22 the left site was treated by topical application of 0.03 ml distilled water while the right side received 0.03 ml 10% w/v M-3175 in distilled water to one site and 3% w/v M-3175 in distilled water to a second site.The doses were absorbed on to 1 cm diameter absorbent patches and covered by an occlusive dressing for 24 hours
Positive control substance(s):
no

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
distilled water
No. with + reactions:
2
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: distilled water. No with. + reactions: 2.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
distilled water
No. with + reactions:
0
Total no. in group:
19
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: distilled water. No with. + reactions: 0.0. Total no. in groups: 19.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
distilled water
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: distilled water. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
distilled water
No. with + reactions:
0
Total no. in group:
19
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: distilled water. No with. + reactions: 0.0. Total no. in groups: 19.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
10% w/v M-3175 in distilled water
No. with + reactions:
1
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 10% w/v M-3175 in distilled water. No with. + reactions: 1.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
10% w/v M-3175 in distilled water
No. with + reactions:
0
Total no. in group:
19
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 10% w/v M-3175 in distilled water. No with. + reactions: 0.0. Total no. in groups: 19.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
10% w/v M-3175 in distilled water
No. with + reactions:
1
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 10% w/v M-3175 in distilled water. No with. + reactions: 1.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
10% w/v M-3175 in distilled water
No. with + reactions:
0
Total no. in group:
19
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 10% w/v M-3175 in distilled water. No with. + reactions: 0.0. Total no. in groups: 19.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
3% w/v M-3175 in distilled water
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 3% w/v M-3175 in distilled water. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
3% w/v M-3175 in distilled water
No. with + reactions:
0
Total no. in group:
19
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 3% w/v M-3175 in distilled water. No with. + reactions: 0.0. Total no. in groups: 19.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
3% w/v M-3175 in distilled water
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 3% w/v M-3175 in distilled water. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
3% w/v M-3175 in distilled water
No. with + reactions:
1
Total no. in group:
19
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 3% w/v M-3175 in distilled water. No with. + reactions: 1.0. Total no. in groups: 19.0.

Any other information on results incl. tables

Induction

Intradermal injection of concertration encorporating FCA caused moderate erythema and occasional cases of eschar formation and pallor at the injection site. Similar injection of 5% w/v M-3175 in distilled water or distilled water alone caused barely perceptible to moderate erythema

Occluded topical application of distilled water to control animals caused no reaction. Similar application of 30% w/vM-3175 in distilled water to test animals caused barely perceptible or slight erythema and exfoliation.

Challenge

Challenge application of distilled water caused slight erythema in two control animals. Exfoliation was also observed in one of these animals. No reaction was observed in test animals.

Challenge application of 10% w/v M-3175 in distilled water caused slight erythema in two control animals. Agreyish/brown discolouration of the test site was also observed in the majority of test and control animals.

Challenge application of 3% w/v M-3175 in distilled water caused slight erythema in one test animal only. Agreyish/brown discolouration of the test site was also observed in one control and four test animals

Bodyweight and general health

All surviving guinea-pigs remained in overt good health and generally made expected overall bodyweight gains during the study. One male test animal (No 323) was killed on human grounds on Day 16. Ante Mortem signs were confined to thin body conformation and bodyweight loss of 61g since Day 1 Necropsy revealed on emaciated build and green/brown staining of the muzzle. This death of this animal was considered incidental to treatment.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
The incidence of significant reactions was similar among test and control group animals challenged with M-3175. Such reactions that were observed were considered to be a result of slight irritation. It was therefore, concluded that, under the conditions of this study, repeated administration of M-3175 did not cause delayed contact hypersensitivity in guinea-pigs.
Executive summary:

The potential of M-3175 to cause delayed contact hypersensitivity in guinea-pigs was assessed by the Magnusson-Kligman Maximisation Test.

The shaven dorsum of each of ten male and ten female Dunkin-Hartley guinea-pigs was subjected to intradermal injections of Freunds Complete Adjuvant, 5% w/v M-3175 in distilled water and 5% w/v M-3175 in the adjuvant on Day 1. Seven days later the same area of skin was treated by topical application of 30% w/v M-3175 in distilled water and the test site was covered by an occlusive dressing for 48 hours. The same induction procedures were carried out on a contemporaneous control group, except that the test material was replaced by vehicle in all doses.

On Day 22 all animals were challenged by occluded topical application of distilled water to the left flank and 10% and 3%w/v M-3175 in distilled water to two sites on the right flank. The occlusive dressings were removed on the following day and the condition of the test sites was assessed approximately 24 -and 48 hours later.

Challenge application of distilled water caused a significant reaction (slight erythema or more marked response) in two control animals. No reaction was observed in test animals.

Challenge application of 10% w/v M-3175 in distilled water caused a significant reaction in two control animals. No reaction was observed in test animals.

Challenge application of 3% w/v M-3175 in distilled water caused a significant reaction in one test animal. No reaction was observed in control animals.

The incidence of significant reactions was similar among test and control group animals challenged with M-3175. Such reactions that were observed were considered to be a result of slight irritation. It was therefore, concluded that, under the conditions of this study, repeated administration of M-3175 did not cause delayed contact hypersensitivity in guinea-pigs.