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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was completed in 2000 according to an OECD guideline and in accordance with GLP. The material is well characterized.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report date:
2002

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
GLP compliance:
yes
Type of study:
mouse local lymph node assay (LLNA)

Test material

Constituent 1
Chemical structure
Reference substance name:
(2S)-2-{[(tert-butoxy)carbonyl]amino}-2-(3-hydroxyadamantan-1-yl)acetic acid
EC Number:
700-361-0
Cas Number:
361442-00-4
Molecular formula:
C17H27NO5
IUPAC Name:
(2S)-2-{[(tert-butoxy)carbonyl]amino}-2-(3-hydroxyadamantan-1-yl)acetic acid
Constituent 2
Reference substance name:
1-Hydroxyadamantanyl-3-(S)-Boc-glycine
IUPAC Name:
1-Hydroxyadamantanyl-3-(S)-Boc-glycine
Details on test material:
white powder; stored at room temperature in the dark; received at testing laboratory on May 7, 2002.

In vivo test system

Test animals

Species:
mouse
Strain:
other: Crl: CBA/Ca Cru,BR
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd
- Age at study initiation: eight to nine weeks old on day 1
- Weight at study initiation: weight range of 15 to 19 grams
- Housing: Up to four female mice were accommodated in suspended polypropylene cages with solid floors and wire mesh lids. Fresh wood flakes were provided as floor litter and this was changed twice weekly.
- Diet (e.g. ad libitum): Free access to mains drinking water and food was allowed throughout the study.
- Water (e.g. ad libitum):Free access to mains drinking water and food was allowed throughout the study.
- Acclimation period: 8 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25 C
- Humidity (%): 40 to 70%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light):12 hours/12 hours


IN-LIFE DATES: From: July 16th To:July 23rd, 2002

Study design: in vivo (LLNA)

Vehicle:
dimethylformamide
Concentration:
Treated with 0.025 ml per ear (pinna) (0.05 mL both ears) of the test solution in dimethyl formamide at concentrations of 10%, 25% and 50% w/w.
No. of animals per dose:
Three groups of four animals each were treated at each concentration group and a further group of four animals were treated as a control group with 1 % m/v Dimethylformamide alone.

Details on study design:
The 3 groups were treated for three days with the test formulation by using a glass syringe to the dorsal surface of each ear. All animals were observed on Day 0, 1, and 2. On Day 6 all mice were placed under an infrared lamp to improve IV dosing and were injected with 0.250 ml of a phosphate buffered saline solution containing tritiated thymidine to the tail vein giving a total of 20 uCi to each mouse. Any signs of toxicity or ill health were noted. Body weights of each mouse were recorded before dosing and before termination. Five hours after administration of tritiated thymidine all mice were killed by halothane asphyxiation and their auricular lymph nodes were excised and pooled for each experimental group. A 5 ml of phosphate buffered saline was added to each set of lymph nodes.The lymph nodes collected into each petri dish were cut open and disaggregated by squashing the fragments with a sharp blade. The resultant liquor was collected into a code-identified conical tube. The petri dish was rinsed with an additional 5 mL phosphate buffered saline and the second liquor was added to the first liquor. At each washing, debris such as connective tissue or fragments of capsule were retained in the petri dish if possible. The pooled liquor was taken off any sediment five minutes after the two liquors had been mixed, placed in a syringe and pushed through a fine mesh in to a conical centrifuge tube to further remove any debris. The liquor was centrifuged at 190 G for 10 minutes. After disposal of the supernatant the pellet was resuspended in 5 mL phosphate buffered saline. This liquor was also centrifuged at 190 G for ten minutes prior to disposal of the supernatant and re-suspension of the pellets in 5% m/v aqueous trichloroacetic acid. These suspensions were stored overnight at approximately 5°C.

On the following day the suspensions were re-centrifuged at 190 G for 10 minutes and the supernatant was drawn off. The pellets were resuspended in 1 mL 5% m/v aqueous trichloroacetic acid. This preparation was subject to ultrasonic dispersion for 25 minutes to ensure a thoroughly dispersed suspension. Prepared suspensions (1 mL) were added to code-identified scintillation pots containing 9 mL Optiphase "Hisafe" scintillation fluid and were mixed by agitation with a Pasteur pipette. Once prepared the scintillation pots were placed in the appropriate carrier racks with an Optiphase blank sample and an Optiphase/5% m/v aqueous trichloroacetic acid (9:1 by volume) blank. The carrier rack was passed into the scintillation counter (Tricarb 2100TR). This operated on a program specific to tritiated samples. The counting period was ten minutes.

Determination of the tritiated thymidine incorporation was completed by measuring radioactive disintegration for each experimental group lymph node cells by using a beta-scintillation counter.
Positive control substance(s):
not specified

Results and discussion

Positive control results:
no data indicated

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: see Remark
Remarks:
Results reported as proliferation index (PI) which is equivalent to a simulation index (SI). Results reported as proliferation index (PI). Concentration of test material the vehicle at 10 % resulted in a proliferation index (PI) of 2.0 which is a negative result. Concentration of test material in the vehicle at 25 % resulted in a proliferation index (PI) of 1.1 which is a negative result. Concentration of test material in the vehicle at 50 % resulted in a proliferation index (PI) of 2.4 which is a negative result.
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: see Remark
Remarks:
Results reported as counts per minute (cpm) which is equivalent to a disintegration per minute (DPM) Results reported as . per minute (cpm) Concentration of test material in the vehicle at 10 % resulted in a mean disintegration of 1405.08 cpm. Concentration of test material in the vehicle at 25 % resulted in a mean disintegration of 806.49 cpm. Concentration of test material in the vehicle at 50 % resulted in a mean disintegration of 1703.35 cpm.

Any other information on results incl. tables

Sample identity

Number of sites yielding lymph nodes

Counts per minute* (CPM)

Counts per minute per node (CLM)

Proliferation Index (PI)

Scintillation fluid (blank)

 

17.34

 

 

Scintillation fluid with 5% m/v trichloroacetic acid

 

104.98

 

 

Vehicle control (dimethylformamide)

8

714.4

89.30

 

BMS 528233-01, 10% m/v

8

1405.08

175.64

2.0

BMS 528233-01, 25% m/v

8

806.49

100.81

1.1

BMS 528233-01, 50% m/v

8

1703.35

212.92

2.4

* All scintillation counts corrected for the blank (except for Scintillation Fluid sample itself)

PI =Test group CLM value

Control group CLM value

TABLE 2 Body weights

Group

Animal

Body weights (g)

Number

number

Day -1

Day 6

1

1

2

3

4

16

19

18

16

17

18

19

19

2

5

6

7

8

15

17

18

17

17

18

19

18

3

9

10

11

12

17

18

18

17

18

20

19

19

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
The Local Lymph Node Assay demonstrated that BMS 528233-01 does not have the potential to cause skin sensitisation.
Executive summary:

This study was conducted to assess the potential of BMS 528233-01 to cause skin sensitisation in the mouse. The test article was prepared for administration at 10, 25 or 50% m/v in a solution dimethylformamide. Groups of four female CBA mice were subjected to topical applications of vehicle or of one of the test formulations to the outer aspect of the auditory pinnae once daily on Days 1, 2 and 3. On Day 6 a 20 µCi dose of tritiated thymidine was injected intravenously into each mouse. Five hours later the auricular lymph nodes were recovered from each animal. The nodes from mice subjected to the same treatment were pooled and suspensions of the cellular components of the lymph nodes were prepared in 5% m/v trichloroacetic acid and processed through a scintillation counter.

Test results are expressed in terms of Proliferation Indices; the ratio of the scintillation count per lymph node obtained from a test group relative to the corresponding scintillation count from controls. The threshold level for the Proliferation Index to be considered a positive indicator of moderate to severe potential to cause skin sensitisation is 3.0.

 

Concentration of test article in applied formulation (%m/v)

10%

25%

50%

Proliferation Index

2.0

1.1

2.4

 

The Local Lymph Node Assay demonstrated that BMS 528233-01 does not have the potential to cause skin sensitisation.