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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

Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vitro
Type of information:
(Q)SAR
Adequacy of study:
key study
Study period:
08 February - 22 March, 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
The objective of this study is to evaluate the ability of PF-07094402 to activate the antioxidant/ electrophile responsive element (ARE)-dependent pathway in the KeratinoSens(TM) assay.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2019

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 442D (In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase Test Method)
GLP compliance:
yes
Justification for non-LLNA method:
The objective of this study is to evaluate the ability of PF-07094402 to activate the antioxidant/ electrophile responsive element (ARE)-dependent pathway in the KeratinoSens(TM) assay.

Test material

Constituent 1
Chemical structure
Reference substance name:
(isopropyl (1s,3s)-3-(methyl(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclobutane-1-carboxylate)
EC Number:
950-860-4
Molecular formula:
C15H20N4O2
IUPAC Name:
(isopropyl (1s,3s)-3-(methyl(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)cyclobutane-1-carboxylate)
Test material form:
solid: bulk
Details on test material:
White Crystalline Solid

In vitro test system

Details on the study design:
Experimental Design

Plating of Cells
For testing, cells were 80-90% confluent. One day prior to testing cells were harvested, and distributed into 96-well plates in basic medium. For each repetition, three replicates were used for the luciferase activity measurements, and one parallel replicate used for the MTT cell viability assay. The cells were incubated overnight in the incubator.

Treatment of Cells
The medium was removed and replaced with fresh culture medium to which 50 µL of the 25-fold diluted test chemical and control items were added. Three wells per plate were left empty to assess background values. The treated plates were covered with foil and then incubated for about 48 hours ± 1 h at 37±1.0oC in the presence of 5% CO2. In total 2 valid experiments were performed.

Luciferase Activity Measurement
The Steady-Glo Luciferase Assay Buffer and Steady-Glo Luciferase Assay Substrate from Promega were mixed together. The assay plates were removed from the incubator and the medium is removed. Then 200 µL of the Steady-Glo Luciferase substrate solution was added to each well. The plates were shaken for at least 5 minutes at room temperature. Plates with the cell lysates were placed in the TECAN Infinite® M200 Pro Plate Reader to assess the quantity of luciferase.

Cytotoxicity Assessment
For the KeratinoSensTM cell viability assay, medium was replaced after the 48 hour exposure time with fresh medium containing MTT and cells were incubated for 3 - 4 hours at 37°C ± 1.0°C in the presence of 5% CO2. The MTT medium was then removed and cells were lysed overnight by adding 10% SDS solution to each well. After shaking, the absorption was measured at 570 nm with the TECAN Infinite® M200 Pro Plate Reader

Results and discussion

In vitro / in chemico

Results
Key result
Parameter:
other: Results from KeratinoSens Assay
Remarks on result:
no indication of skin sensitisation

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
In conclusion, PF-07094402 is classified as negative (no activation of the antioxidant/electrophile responsive element (ARE)-dependent pathway in keratinocytes) under the experimental conditions described in this report.