4-(1-piperidinyl)-2-(E)-butenoicacid hydrochloride

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28th November 2011 to 6th December 2011

Reliability:

1 (reliable without restriction)

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:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

1-[(2E)-3-carboxyprop-2-en-1-yl]piperidin-1-ium chloride

EC Number:

690-933-5

Cas Number:

197892-69-6

Molecular formula:

C9H16ClNO2

IUPAC Name:

1-[(2E)-3-carboxyprop-2-en-1-yl]piperidin-1-ium chloride

Test animals

Species:

rat

Strain:

Wistar

Details on species / strain selection:

Recognized by international guidelines as the recommended test system (e.g. EPA, FDA, OECD and EC).

Sex:

male/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: Approximately 10 weeks.
- Housing: Room Number A0.03.
Group housing of 3 animals per sex in Makrolon cages (MIV type, height 18 cm) with sterilized sawdust as bedding material and paper as cage-enrichment.
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Tap water ad libitum
- Acclimatisation period: At least 5 days before the start of treatment under laboratory conditions.

DETAILS OF FOOD AND WATER QUALITY:
Food: Results of analyses for nutrients and contaminants were examined and archived.
Water: Certificates of analysis (performed quarterly) were examined and archived.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.6 – 21.2ºC
- Humidity (%): 43 - 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: 5th December 2011 To: 13th December 2011

Administration / exposure

Route of administration:

oral: gavage

Details on route of administration:

Dose Volume:
5 mL/kg body weight.
Actual dose volumes were calculated according to body weights determined on Day 1 (dosing on Days 1-4) and on Day 4 (dosing on Days 5-7).

Vehicle:

water

Remarks:

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

Details on oral exposure:

Oral gavage, using a plastic feeding tube.
Formulations were placed on a magnetic stirrer during dosing.

Duration of treatment / exposure:

7 days

Frequency of treatment:

Once daily for up to 7 consecutive days, approximately the same time each day with a maximum of 6 hours difference between the earliest and latest dose. Animals were dosed up to the day prior to necropsy.

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

150 mg/kg bw/day (actual dose received)

Doses / concentrations 2

Dose / conc.:

300 mg/kg bw/day (actual dose received)

Doses / concentrations 3

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

Three

Control animals:

yes, concurrent vehicle

Details on study design:

Observations
Mortality / Viability: At least twice daily.
Clinical signs: At least once daily from start of treatment onwards, detailed clinical observations were made in all animals. The time of onset, grade and duration of any observed signs were recorded. Signs were graded for severity and the maximum grade was predefined at 3 or 4. Grades were coded as slight (grade 1), moderate (grade 2), severe (grade 3) and very severe (grade 4). For certain signs, only its presence (grade 1) or absence (grade 0) was scored.
Body weights: On Days 1, 4 and 7.
Food consumption: Over Days 1-4 and 4-7.
Water consumption: Subjective appraisal was maintained during the study, but no quantitative investigation introduced as no effect was suspected.

Clinical laboratory investigations
Blood samples were collected under anaesthesia using isoflurane, between 7.00 and 10.30 a.m. on the day of necropsy. Animals were deprived of food overnight (with a maximum of 20 hours), but water was available. Blood samples were drawn from the retro-orbital sinus of all rats/sex/group and collected into tubes prepared with EDTA for haematological parameters (0.5 mL), with citrate for clotting tests (0.45 mL) and Li-heparin treated tubes for clinical biochemistry parameters (0.5 mL). An additional blood sample (0.25 mL) was collected into untreated tubes for determination of bile acids.

Examinations

Sacrifice and pathology:

Necroscopy
Animals were deeply anaesthetized using isoflurane and subsequently exsanguinated and subjected to a full post mortem examination. Animals were deprived of food overnight prior to scheduled necropsy. Descriptions of all macroscopic abnormalities seen at post mortem were recorded.
Samples of the following tissues and organs were collected from all animals at necropsy and fixed in 10% buffered formalin (neutral phosphate buffered 4% formaldehyde solution, adrenal glands, ovaries, brain, heart, kidneys, liver, spleen, stomach, testes, thymus and all gross lesions.

Statistics:

Observations/measurements in the study were recorded electronically using the following programs:
- REES Centron Environmental Monitoring system version SQL 2.0 (REES Scientific, Trenton, NJ, USA): Environmental monitoring.
- TOXDATA version 8.0 (NOTOX B.V., ‘s-Hertogenbosch, The Netherlands): Mortality / Clinical signs / Body weights / Food consumption / Organ weights.
- ADVIA® 120 Hematology SystemVersion V.3.1.8.0.MS (Siemens Healthcare Diagnostics B.V., Breda, The Netherlands): Haematology.
- STA Compact® version 1.06.06 (Diagnostica Stago S.A.S., Asnières, France): Clotting parameters.
- AU400® Chemistry System version 8.2A (Beckman Coulter Nederland B.V., Woerden, The Netherlands): Clinical biochemistry.
- Pathdata version 6.2D (Pathology Data Systems, Basel, Switzerland): Histopathology

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Body weights and body weight gain of treated animals remained in the same range as controls over the study period.
Temporary slight weight loss, observed for individual animals of all dose groups is more often seen in rats of this age. In addition, since the incidence of the slight weight loss was unrelated to the dose,
this finding was considered to be of no toxicological significance.

Food efficiency:

no effects observed

Haematological findings:

no effects observed

Description (incidence and severity):

No toxicologically relevant changes occurred in haematological parameters of treated rats.
Individual increases of neutrophil counts with concurrently reduced lymphocyte counts were noted among the dose groups without a treatment related distribution. This shift in type of white blood cells was considered to be a secondary non-specific response to stress and to be of no toxicological significance.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

No toxicologically relevant changes occurred in clinical biochemistry parameters of treated rats.
Higher bile acids levels among treated animals were considered to have arisen as a result of slightly low control values and in the absence of a treatment-related incidence were considered to be of no toxicological significance.

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

No toxicologically relevant changes were noted regarding organ weights and organ to body weight ratios.
The reduced size of the left testis of one control male was reflected in the relatively low weight of the testes. This finding is occasionally seen among rats used in these types of studies and is considered a change of no toxicological significance.

Gross pathological findings:

no effects observed

Description (incidence and severity):

Macroscopic observations at necropsy did not reveal any alterations that were considered to have arisen as a result of treatment.
Incidental findings included a yellowish soft nodule on the tail of the right epididymis of two control males, reduced size of the left testis and left epididymis of one control male, an enlarged spleen of one male at 300 mg/kg, a watery-clear cyst on the right kidney of one female at 150 mg/kg and presence of fluid in the uterus of two control females, one female at 150 mg/kg and one female at 300 mg/kg. These findings are occasionally seen among rats used in these types of studies and are considered changes of no toxicological significance.

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical biochemistry

clinical signs

food consumption and compound intake

gross pathology

haematology

mortality

organ weights and organ / body weight ratios

Remarks on result:

not determinable due to absence of adverse toxic effects

Target system / organ toxicity

Key result

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

From the results presented in this report, it was concluded that PF-00818977-01 has no toxic potential when administered to rats by daily oral gavage for a period of 7 consecutive days up to 1000 mg/kg.

Executive summary:

Title: 7-Day repeated dose study with PF-00818977-01 by daily gavage in the rat.
Guidelines
The study was based around the following guidelines:
- EC No 440/2008, B.7 Repeated Dose (28 days) Toxicity (oral), 2008.
- OECD 407, Repeated Dose 28-day Oral Toxicity Study in Rodents, 2008
Rationale for dose levels
Dose levels for this 7-day oral gavage study were selected to be 0, 150, 300 and 1000 mg/kg, based on the results of the acute oral toxicity study (NOTOX project 497144) and in consultation with the sponsor.
Study outline
The test substance, formulated in water, was administered daily for 7 consecutive days by oral gavage to SPF-bred Wistar rats. One control group and three treated groups were tested, each consisting of 3 males and 3 females.
Evaluated parameters
The following parameters were evaluated: clinical signs daily; body weight on Days 1, 4 and 7 and food consumption over Days 1-4 and 4-7; clinical laboratory investigations and macroscopy at termination; organ weights and histopathology on a selection of tissues.
Results
No toxicologically significant changes were noted in any of the following parameters investigated in this study: clinical appearance, body weight, food consumption, clinical laboratory investigations, macroscopic examination, organ weights, and microscopic examination.
Conclusion
From the results presented in this report, it was concluded that PF-00818977-01 has no toxic potential when administered to rats by daily oral gavage for a period of 7 consecutive days up to 1000 mg/kg.