D_Attch 1. Statement of Work.pdf

Version 11-30-2016-1 FAR Part 11—Describing Agency Needs

STATEMENT OF WORK (SERVICES)

(SOW)

GENERAL INFORMATION

Title of Project:

In vivo testing of a lead CoQ10 biosynthesis inhibitor in an SK-BR-3 xenograft mouse model

Statement of Need and Purpose:

NCATS TDB is requesting this service to assess our lead small molecule ColQ10 biosynthesis inhibitor in an SK-BR-3 xenograft mouse model.

Background Information and Objective:

NCATS TDB has been collaborating with a team at Columbia Univ. to develop small molecules inhibitors of CoQ10 biosynthesis for the treatment of cancer. Currently, we have a series of inhibitors that show good potency and selectivity in vitro. We would like to test our lead inhibitor in a mouse model xenografted with SK-BR-3 human breast cancer cells. NCATS does not have in vivo efficacy testing facility nor capacity to run in vivo efficacy models, and therefore must get this service through a contractor.

Period of Performance:

12 months

SCOPE OF WORK

General Requirements:

Independently and not as an agent of the Government, the Contractor shall furnish all the necessary services, qualified personnel, material, equipment, and facilities, not otherwise provided by the Government as needed to perform the Statement of Work below:

Specific Requirements:

The in vivo studies consist of a series of 4 studies. Study 1 will assess the PK properties of the lead inhibitor in SKBR3 xenografted mice, Study 2 will assess the 14 day toxicology of the lead inhibitor in NCG mice, Study 3 will assess acute efficacy in SKBR3 xenografted mice, and Study 4 will assess 21 day efficacy of lead inhibitor in xenogragted mice. Studies 2-4 are listed as optional, and study initiation will be triggered by successful completion and promising results from previous studies.

Study #1: Pharmacokinetic (PK) properties of lead inhibitor in SKBR3 tumor bearing NCG mice.

Timeline: 2-3 weeks

Mice per group: 4

Total mice: 6timepoints X 1modes of administration X 2doses X 4mice per group + 4 micetime 0 = 52 (+ 13 extra) = 65

SKBR3 cells will be xenografted into NCG mice by injecting 10 million cells in 100% Matrigel to 2 sites: the mammary gland area and flank area. When tumors size reaches an average of 80mm3, mice will be divided into groups (keeping average tumor size even between groups and compound will be administered at 20 and 40 mg/kg via

IP.

Plasma and tumor samples will be collected at 6 timepoints after administration: 0.5, 1, 2, 4, 8 and 24 hours. Additional samples will be collected from 4 untreated mice (time 0). Samples will be stored at -80 oC and shipped to study sponsors.

Study #2 (Optional): Toxicity study for lead inhibitor in NCG mice.

Timeline: 2 weeks

Mice per group: 3

Total mice: 6

Lead inhibitor will be tested for general toxicity by daily IP injection at a dose determined based on study 1 results.

Treated mice will be compared to a control group that will receive vehicle injections only.

Clinical signs of toxicity will be monitored and recorded daily: weight loss, reduced food/drink consumption, labored breathing, and death.

One day after final injection (day 15, see Fig 2), the mice will be euthanized. Tissues (brain, lung, heart, liver, spleen, pancreas, intestine and kidney) will be collected, frozen under liquid nitrogen, stored at -80 oC and shipped to study sponsor.

Study 3 (Optional): Acute efficacy study to quantify CoQ10 levels in SKBR3 tumor-xenografts.

Figure 1

Day 1

SKBR3

cells injection

~Day 14

Tumors reach ~ 80 mm3:

Divide into groups and administer compound

30 min 1 h

2 h 4 h

Tumors and plasma collection

Timeline

8 h 24 h

Figure 2

To ensure that the lead inhibitor induced a reduction in CoQ10 abundance in tumors in vivo, xenografted tumors will be harvested to quantify CoQ10 (reduced and oxidized) levels after short treatment with our lead inhibitor.

Timeline: 3-4 weeks

Mice per group: 4

Total mice: 8 (+ 2 extra)

SKBR3 cells will be injected as in Study 1. Xenografts will be allowed to grow until tumors reach 100 mm3 in diameter and then the mice will be randomly distributed into 2 groups [4 mice /group].

Group 1: daily IP injection with 100 microliters of the vehicle Group 2: daily IP injection with lead inhibitor compound at xx mg/kg

On the 4th day, 6 hours after the 4th injection, the mice will be euthanized and tumors will be collected, flash frozen and transferred to study sponsor.

Study #4 (Optional): Efficacy study for lead inhibitor in SKBR3 tumor-xenografts.

Timeline: 4-5 weeks.

Mice per group: 10

Total mice: 40 (+ 10 extra)

SKBR cells will be xenografted into NCG mice as in studies 1 and 3.

Once tumors reached 100 mm3 in diameter, the mice will be randomly distributed into 4 groups [10 mice /group].

Group 1: mice will receive daily IP injection with 100 microliters of the vehicle.

Group 2: mice will receive daily IP injection with 100 microliters of lead inhibitor at xx mg/k.

Group 3: mice will receive daily IP injection with 100 microliters of lead inhibitor at xx mg/kg.

Group 4: mice will receive daily IP injection with 100 microliters of positive control at xx mg/kg.

Tumor size and body weight will be monitored for the duration of the 21 day injection period.

Figure 3

SKBR3

cells injection

Day 1

Tumors reach ~ 100 mm3:

Divide into groups

Tumors collection

Timeline

6h

2-3 weeks

Day 2 Day 3 Day 4 inject compound

Figure 4

SKBR3

cells injection

Day 0

Tumors reach ~ 100 mm3:

Divide into groups

Tumors collection

Timeline

2-3 weeks inject compound daily

Day 21

On day 21 (Fig 4), the mice will be euthanized. Tumors, and tissues (brain, lung, heart, liver, spleen, pancreas, intestine and kidney) will be collected, frozen under liquid nitrogen, stored at -80 oC and shipped to study sponsor.

LEVEL OF EFFORT:

PhD level scientists with knowledge and training in in vivo animal model testing shall conduct/direct experiments.

GOVERNMENT RESPONSIBILITIES

NCATS will ship lead inhibitor at sufficient quantities to CRO for testing.

DELIVERY OR DELIVERABLES

1. The final report shall contain raw data, data tables and graphs.

DELIVERABLES DELIVERY DATE

Draft Study Report with raw data, data tables and graphs

6 weeks after completion of each study

Final Study Report with raw data, data tables and graphs

8 weeks after completion of each study

2. Tumors or tissues collected as listed in each study.

REPORTING REQUIREMENTS

The vendor shall send a full report with raw data. Specifically, the following shall be included: Title page, brief introduction/goals of the study, materials and methods, results summary, discussion, appendix, including data/graphs.

OTHER CONSIDERATIONS

Travel:

N/A

Key Personnel:

Information System Security Plan:

Data Rights:

Section 508—Electronic and Information Technology Standards:

Publications and Publicity:

Confidentiality of Information: