PF-05201372

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23rd August 2011 to 26th January 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nohsan, Notification No. 8147, April 2011; including the most recent partial revisions.

GLP compliance:

yes

Test type:

acute toxic class method

Test material

Test material information

Constituent 1

Reference substance name:

(2E)-N-(5-Cyano-4-{(E)-[(dimethylamino)methylene]amino}-2-methoxyphenyl)-4-(piperidin-1-yl)but-2-enamide

Molecular formula:

C20 H27 N5 O2

IUPAC Name:

(2E)-N-(5-Cyano-4-{(E)-[(dimethylamino)methylene]amino}-2-methoxyphenyl)-4-(piperidin-1-yl)but-2-enamide

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Females: nulliparous and non-pregnant
- Age at study initiation: approx. 9-12 weeks old
- Weight at study initiation: Body weight variation did not exceed +/- 20% of the sex mean.
- Fasting period before study:
- Housing: Group housing of 3 animals per cage in labeled Makrolon cages (MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water
- Acclimation period: at least 5 days before start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.3 – 21.9ºC
- Humidity (%): 40 - 59%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day.

IN-LIFE DATES: From: 09 November 2011 To: 30 November 2011

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Elix, Millipore S.A.S., Molsheim, France

Details on oral exposure:

The formulations (w/w) were prepared within 4 hours prior to dosing.
Homogeneity was accomplished to a visually acceptable level. No correction was made for purity of the test substance. The concentration of the test substance in vehicle was varied to allow constant dosage volume in terms of mL/kg body weight.

Doses:

2000 mg/kg (10 mL/kg) body weight.
300 mg/kg (10 mL/kg) body weight.

No. of animals per sex per dose:

stepwise treatment of groups of 3 females

Details on study design:

Observations
Mortality/Viability: Twice daily.
Body weights: Days 1 (pre-administration), 8 and 15.
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15. The symptoms were graded according to fixed scales and the time of onset, degree and duration were recorded:
Maximum grade 4: grading slight (1) to very severe (4)
Maximum grade 3: grading slight (1) to severe (3)
Maximum grade 1: presence is scored (1).
Necropsy: The moribund animal and animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities recorded.

Statistics:

Observations/measurements in the study were recorded electronically using the following programmes: REES Centron Environmental Monitoring system version SQL 2.0 (REES scientific, Trenton, NJ, USA); TOXDATA version 8.0 (NOTOX B.V., ‘s-Hertogenbosch, The Netherlands): Clinical signs, Body weights.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

500 mg/kg bw

Based on:

test mat.

Mortality:

All animals at 2000 mg/kg were found dead or sacrificed in moribund condition on Day 1. No mortality occurred among the animals at 300 mg/kg.

Clinical signs:

irregular respiration

lethargy (hypoactivity)

observations of tremors

other:

Body weight:

other body weight observations

Remarks:

The body weight gain shown by the animals at 300 mg/kg over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.

Gross pathology:

No toxicologically significant abnormalities were found at macroscopic post mortem examination of the animals.
One animal at 300 mg/kg showed an enlarged mandibular lymph node, which is occasionally seen among rats of this age and strain and was not observed for the animals at 2000 mg/kg. Therefore, this finding was considered not toxicologically significant.

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The oral LD50 value of PF-05201372 in Wistar rats was established to be within the range of 300- 2000 mg/kg body weight.
According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 500 mg/kg body weight.

Executive summary:

Assessment of acute oral toxicity with PF-05201372 in the rat (Acute Toxic Class Method).
The study was carried out based on the guidelines described in:
OECD No.423 (2001) "Acute Oral Toxicity, Acute Toxic Class Method" Commission Regulation (EC) No 440/2008, B1 tris: "Acute Oral Toxicity, Acute Toxic Class Method" EPA,

OPPTS 870.1100 (2002), "Acute Oral Toxicity"
JMAFF guidelines (2011) including the most recent partial revisions.
Initially, PF-05201372 was administered by oral gavage to three female Wistar rats at 2000 mg/kg body weight. In a stepwise procedure two additional groups of females were dosed at 300 mg/kg body weight. All animals were subjected to daily observations and weekly determination of body weight.
Macroscopic examination was performed on the day of death or after terminal sacrifice (Day 15).
All animals at 2000 mg/kg were found dead or sacrificed in moribund condition on Day 1. No mortality occurred among the animals at 300 mg/kg. All animals at 2000 mg/kg showed the following symptoms prior to death on Day 1: lethargy, general tremors, flat and/or hunched posture and/or shallow respiration.
All animals at 300 mg/kg showed hunched posture and piloerection on Day 1. In addition, hunched posture was also observed for all animals on Day 2 and for three animals on Day 3. The body weight gain shown by the animals at 300 mg/kg over the study period was considered to be normal.
No toxicologically significant abnormalities were found at macroscopic post mortem examination of the animals.
The oral LD50 value of PF-05201372 in Wistar rats was established to be within the range of 300-2000 mg/kg body weight.
According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 500 mg/kg body weight.
Based on these results:
- according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2011), PF-05201372 should be classified as: harmful if swallowed (Category 4) for acute toxicity by the oral route.
- according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures, PF-05201372 should be classified as Category 4 and should be labeled as H302: Harmful if swallowed.