(3R,6S)-1-benzyl-6-methylpiperidin-3-amine acetyl-L-leucinate [1:1]

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21st February 2020 to 16th April 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

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:

other: EC No 440/2008, part B: "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: JMAFF Guidelines (2000), including the most recent revisions.

GLP compliance:

yes

Test type:

acute toxic class method

Test material

Test material information

Constituent 1

Reference substance name:

(3R,6S)-1-benzyl-6-methylpiperidin-3-amine acetyl-L-leucinate [1:1]

EC Number:

840-203-9

Cas Number:

2295746-51-7

Molecular formula:

C21H35N3O3

IUPAC Name:

(3R,6S)-1-benzyl-6-methylpiperidin-3-amine acetyl-L-leucinate [1:1]

Test material form:

solid: particulate/powder

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: Yes
- Age at study initiation: approximately 9-10 weeks old
- Weight at study initiation: 153 to 196 g.
- Fasting period before study: Animals were deprived of food overnight (for a maximum of 20 hours) prior to dosing and until 3-4 hours after administration of the test item. Water was available.
- Housing: On arrival and following assignment to the study, animals were group housed (up to 3 animals of the same sex and same dosing group together) in polycarbonate cages (Makrolon MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier &Söhne GmbH+ CO. KG, Rosenberg, Germany) equipped with water bottles. Animals were separated during designated procedures/activities. Each cage was clearly labeled.
- Diet: Pelleted rodent diet(SMR/M-Z from SSNIFF® Spezialdiäten GmbH,Soest, Germany) ad libitum
- Water: Municipal tap-water ad libitum
- Acclimation period: at least 5 days before the commencement of dosing.
- Method of randomisation in assigning animals to test and control groups: Animals were assigned to the study at the discretion of the coordinating biotechnician.

ENVIRONMENTAL CONDITIONS
- Temperature: 20 - 21°C
- Humidity: 36 - 52%
- Air changes: Ten or greater air changes per hour with 100% fresh air (no air recirculation) were maintained
- Photoperiod: 12hrs dark / 12 hrs light

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

propylene glycol

Details on oral exposure:

VEHICLE
- Amount of vehicle: A dose volume of 10 mL/kg body weight was used for each dose.

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg

- Rationale for the selection of the starting dose: The dose levels were based on the OECD test guidelines and were selected from the series 5 (lowest dose level), 50, 300 and 2000 (highest dose level) mg/kg body weight. The starting dose level should be the one that is likely to produce mortality in at least some of the animals and was selected based on available toxicity data of the test item.

Doses:

300 and 2000 mg/kg

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 3 times on the day of dosing and once daily thereafter. Animals were weighed individually on Day 1 (pre-dose), 8 and 15. A fasted weight was
recorded on the day of dosing. Terminal body weights were collected from animals found
dead or euthanized moribund after Day 1.
- Necropsy of survivors performed: yes
All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities were recorded.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

>= 300 - <= 2 000 mg/kg bw

Based on:

test mat.

Mortality:

At 2000 mg/kg, all animals were found dead or sacrificed in moribund condition on Days 1 or 2 post-treatment.
At 300 mg/kg, no mortality occurred.

Clinical signs:

other: At 2000 mg/kg, clinical signs consisted of hunched posture, erected fur, abnormal respiratory rate (decreased and/or increased), partly closed eyes, moderate salivation, decreased activity and non-sustained convulsions. At 300 mg/kg, hunched posture in a

Gross pathology:

At 2000 mg/kg, abnormalities of the stomach (abnormal content green fluid/distended with gas), large intestine cecum (abnormal content distended with gas), large intestine colon (abnormal content distended with gas) and small intestine ileum (abnormal content frothy dark yellow) were found in the animals at macroscopic postmortem examination. One incidental finding included an abnormality of the thymus (focus dark black multifocal). This finding is more often seen in rats of this strain and in this type of study and was therefore considered to be not toxicologically relevant.

At 300 mg/kg, macroscopic postmortem examination at termination did not reveal any
abnormalities.

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Executive summary:

The objective of this study was to assess the toxicity of the test item when administered in a single dose to female rats at one or more defined dosages. Furthermore, the results of the study allowed the test item to be ranked according to most classification systems, currently in use. This study should provide a rational basis for risk assessment in man.

Initially, PF-07202371-S7 was administered by oral gavage to three female Wistar Han rats at 300 mg/kg body weight. In a stepwise procedure two additional groups of three females were dosed at 2000 and 300 mg/kg body weight, respectively. 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).
At 2000 mg/kg, all animals were found dead or sacrificed in moribund condition on Days 1 or 2 post-treatment. At 300 mg/kg, no mortality occurred.
At 2000 mg/kg, clinical signs consisted of hunched posture, erected fur, abnormal respiratory rate (decreased and/or increased), partly closed eyes, moderate salivation, decreased activity and non-sustained convulsions. At 300 mg/kg, hunched posture in all six animals between Days 1 and 3 and erected fur in three animals on Day 1 were noted.
The body weight gain shown by the surviving animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain. At 2000 mg/kg, abnormalities of the stomach (abnormal content green fluid/filled with gas), cecum (abnormal content filled with gas), colon (abnormal content filled with gas) and ileum (abnormal content frothy dark yellow) were found in the animals at macroscopic postmortem examination. At 300 mg/kg, macroscopic postmortem examination at termination did not reveal any abnormalities.
The oral LD50 value of PF-07202371-S7 in Wistar Han 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 (2017) (including all amendments), PF07202371-S7 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 items and mixtures (including all amendments), PF-07202371-S7 should be classified as Category 4 and should be labeled as H302: Harmful if swallowed.