Sample_Task_Order__1_-_Protocol_REV_12-21-15.pdf

SAMPLE PROTOCOL

TASK ORDER #1

PHARMACOKINETICS AND ORAL BIOAVAILABILITY OF [NSC TBD] IN MALE MICE

SPONSOR: Toxicology and Pharmacology Branch

Developmental Therapeutics Program Division of Cancer Treatment and Diagnosis National Cancer Institute National Institutes of Health Bethesda, Maryland 20892

COR: TBD

CONTRACT NUMBER: TBD

CONTRACTOR: TBD

PROJECT MANAGER: TBD

PROJECT DIRECTOR: TBD

PROPOSED IN-LIFE PHASE:

Start: TBD

Finish: TBD hudakad Typewritten Text hudakad Typewritten Text RFP Number: N02CM67000-11 hudakad Typewritten Text hudakad Typewritten Text hudakad Typewritten Text Attachment 19-1P

I. OBJECTIVE

The objective of this protocol is to characterize the plasma pharmacokinetics and oral bioavailability of NSC TBD in male mice.

II. MATERIALS AND METHODS

A. Test Article and Vehicle:

1. Name of Test Article:

NSC TBD

2. Name of Vehicle:

TBD

3. Characterization and Documentation of Methods of Synthesis, Fabrication or Derivation:

a. Test Article:

Compound identity, strength, quality, stability and purity as well as documentation of methods of synthesis, fabrication or derivation are the responsibility of the NCI. Sufficient quantity of drug shall be reserved for archiving from each lot and shipment used.

b. Vehicle:

The vehicles are stable throughout the date of expiration on the container labels when stored according to the manufacturer’s instructions.

B. Test System:

1. Species, Strain Supplier and Test System Justification:

CD-1 mice or other acceptable strain obtained from an approved commercial supplier will be used in this evaluation. This is an accepted species to support studies of compounds used or intended for use in humans.

2. Initial Age, Sex and Weight:

On the first day of dosing, the weight ranges of mice will be approximately 18 to 22 grams and the mice will be approximately 5 to 7 weeks of age. Only males will be used.

3. Care and Housing:

General procedures for animal care and housing will be in accordance with The Guide for the Care and Use of Laboratory Animals, National Research Council, 2011 and the U.S.

Department of Agriculture through the Animal Welfare Act (7 USC

2131), 1985 and Animal Welfare; Standards incorporated in 9 CFR Part 3, 1991. Appropriate caging and bedding or cage board (not cedar or pine chips) will be used. No contaminants shall be present in the bedding which could interfere and affect the results of the evaluation. Environmental parameters will be set to maintain conditions specified in the facility SOPs. Environmental conditions will be within specified limits of at least 90 percent of scheduled observations.

4. Diet and Water Supply:

Diet is to be certified, commercial, dry rodent chow provided ad libitum. Water source will be the public supply given ad libitum. No contaminants will be present in the feed or water which could interfere and affect the results of the evaluation.

5. Quarantine:

All mice will be quarantined for a minimum of 5 days prior to baseline measurements. No prophylactic or therapeutic treatment will be administered during the quarantine period. Only healthy animals will be placed on test.

6. Animal Identification:

All mice will be given a unique identification number for this evaluation by ear tag or other approved method.

C. Experimental Design

1. Randomization:

In order to obtain groups that are comparable by weight, all mice will be randomly assigned from the colony to each treatment group using a computer-based body weight stratification procedure.

2. Group Assignments:

After randomization, 12 male mice will be assigned to each of two dose groups as follows:

Group Cpd. Route Dose

(mg/kg)

Conc. in Formulation

(mg/mL)

Dose Volume (mL/kg) Sub-group No. of

Animals

Blood (Plasma)

Collection Timepoint

(min)

1 NSC TBD IV TBD TBD 5

A 3 5, 120 B 3 15, 240 C 3 30, 480 D 3 60, 1440

2 NSC TBD PO TBD TBD 5

A 3 15, 240 B 3 30, 360 C 3 60, 480 D 3 120, 1440

3. Route of Administration and Reason for Choice:

The test compound will be given as a single intravenous or oral dose in order to characterize its pharmacokinetics and bioavailability in mice.

4. Formulation Preparation:

To be specified for iv and oral administration to mice.

5. Dosing Procedure:

On Day 1, each mouse in a given dose group will receive formulated test article either by iv injection (tail vein) or oral gavage.

The dose volume will be 5 mL/kg of body weight. The amount of drug administered to each mouse will be based on each animal’s most recent individual body weight.

6. Measurements:

a. Clinical signs:

The mice will be observed during the blood sampling period and any adverse clinical signs recorded.

b. Body Weight:

Individually weigh all mice prior to dosing on Day 1.

c. Sample Collection for Plasma Drug Levels:

Whole blood samples (target volume of 200 µL) will be collected from 4 cohorts of 3 mice each from each group via the retro-orbital sinus (or other proposed method). Mice in each cohort will be stagger-sampled twice to obtain the eight timepoints indicated below and in the table above. Mice will be anesthetized and blood will be collected according to established laboratory SOPs. Blood will be placed into tubes containing the anticoagulant EDTA.

Target blood collection time points:

Group 1 (iv): 5, 15, 30, 60, 120, 240, 480, and 1440 minutes after dosing.

Group 2 (oral): 15, 30, 60, 120, 240, 360, 480, and 1440 minutes after dosing.

Actual collection times will be recorded.

Blood samples will be placed on wet ice until centrifuged to obtain plasma. Plasma samples will be transferred into appropriately labeled tubes and placed on dry ice until stored in a freezer set to maintain at least -70 ºC.

e. Necropsy Procedure:

Necropsy is not required in this protocol.

d. Bioanalytical and Pharmacokinetic Analysis:

A bioanalytical method for the compound in mouse plasma will be implemented using standard LC-MS/MS approaches.

Analytical sensitivity shall be as great as possible (ideally < 10 ng/mL). Full validation to FDA guidelines is not required for this analysis.

Total number of samples for quantification of test article in plasma:

8 timepoints x 3 mice/timepoint x 2 groups = 48 samples

Pharmacokinetic data analysis shall include (at a minimum) standard non-compartmental modeling to determine Cmax(po), C0(iv), Tmax, AUC0∞

, AUClast, CL, and t½, Vss, and %F.

III. QUALITY ASSURANCE

A. Type of Evaluation This evaluation will not require compliance with the FDA Good Laboratory Practice Regulations.

B. Standard Operating Procedures

All operations pertaining to this evaluation, unless specifically defined in this protocol, will be performed according to the standard operating procedures of the laboratory and any deviations will be documented.

C. Protocol Amendments

All changes in or revisions of an approved protocol and the reasons therefore will be documented, signed, and dated by the Project Director/Project Manager and the NCI COR. Amendments will be maintained with the protocol. Verbal approval for changes in the protocol may be granted by the NCI COR, but a written amendment will follow.

IV. REPORTING AND DISCUSSION OF DATA

A. Interim Tabulated Data Concentration-time data and derived PK parameters for the test compound shall be provided to the COR in a spreadsheet format, along with suitable c vs. t plots within two weeks of receipt of the compound.

B. Final Report

The results obtained under this protocol will be submitted as a draft report, due two weeks after completion of the evaluation. This report will accurately and completely describe the experimental design, procedures and findings, present an analysis and summary of the data followed by the conclusions derived from the analyses.

The report will also include: (a) a cover page which will include the title, contract number, authors, laboratory address, dates of initiation and completion, and sponsor; (b) an abstract to be placed at the beginning of the final report; (c) a comprehensive summary to be placed after the abstract; (d) the signature of the Project Director/Project Manager and any others deemed necessary; (e) a table of contents. The draft report will be reviewed by the COR and any requests for revisions will be sent to the Project Director/Project Manager. The final report will be due two weeks after return of the draft report for revision.

Protocol Approvals:

Project Director:

(Date)

Project Manager:

(Date)

NCI COR:

(Date)