D.CSS_Attach1-SOW.pdf

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

STATEMENT OF WORK (SOW)

GENERAL INFORMATION

Title of Project:

Bioanalytical Assay Development and Concentration Determination for Remdesivir, GS-441524, GS-441524 Mono- and Tri-Phosphates in Cell Lysates

Statement of Need and Purpose:

The objective of this work is to develop bioanalytical assays for Remdesivir, GS-441524, and GS-441524 mono- and tri-phosphates, and to apply these assays to determine concentrations of Remdesivir, GS-441524, GS-441524 mono- and tri-phosphates in cell lysates collected from in vitro incubations.

Background Information and Objective:

To support drug development for COVID-19 treatment, bioanalytical assays for Remdesivir, parent drug GS-441524, and GS-441524 mono- and tri-phosphates are needed. These assays will be used to quantify the prodrug, parent drug and phosphate metabolite concentrations in cell lysates collected from in vitro incubations with Vera E6, Calu3, HEK293 cells, the cell lines commonly used for in vitro antiviral activity assessments.

Period of Performance:

One hundred and eighty (180) days from the award date.

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:

The Contractor, or Contract Research Organization (CRO), lab should have extensive experience in bioanalytical method development for nucleoside analogs and the phosphate metabolites. Phosphate metabolites of nucleoside analogs are very polar and extremely unstable; therefore, past experience with assay development and sample handling for these types of molecules is very important for the success of this work.

Specific Requirements:

The CRO lab should have solid scientific knowledge on bioanalytical method development for nucleoside analogs and the polar phosphate metabolites. Specific tasks include:

1. Method Development of a 4-in-1 LC-MS/MS assay for Remdesivir, GS-441524, and GS- 441524 mono- and tri-phosphates in cell lysate (or lysis buffer if applicable). Stability of all analytes in the cell lysate will be evaluated as part of Method Development. A solution which stabilizes all analytes will be identified to maintain the stability of analytes in the cell lysate solution

• Chromatography method k’ ≥ 2 for each analyte

• Baseline separation of all analytes and monitor GS-441524 diphosphate SRM

• The liquid chromatography conditions used must have, as a minimum, the following specifications: column inner diameter ≤ 2.1 mm, column stationary phase particle size ≤3. 5 micrometers.

2. Method Qualification of a 4-in-1 LC-MS/MS assay for Remdesivir, GS-441524, and GS- 441524 mono- and tri-phosphates in cell lysate (or lysis buffer if applicable):

• 2 day linearity, precision/accuracy (Minimum of 6 standard concentrations (n=2) and 4 QC concentrations (n=6), including the LLOQ QC)

• Absolute recovery (LQC and HQC, n=5)

• Carryover evaluation.

• Stability (On-ice- minimum of 6 hours and Freeze/thaw-4 cycles at -70°C, LQC and

HQC)

• Acceptance criteria- + 20% difference from nominal concentration at all tested concentrations

3. Sample Analysis

• Remdesivir incubation- 117 samples of cell lysates to measure Remdesivir, GS-

441524, and GS-441524 mono- and tri-phosphates

• GS-441524 incubation- 117 samples of cell lysates to measure, Remdesivir, GS-

441524, and GS-441524 mono- and tri-phosphates

• Repeat the assays on selected samples when needed.

The Contractor will need the following materials as reference and internal standards to perform the assay development and sample analysis described in the specific tasks above:

1. 2 mg stable isotope labeled remdesivir (Internal standard for the assay)

2. 2 mg stable isotope labeled GS-441524 (Internal standard for the assay)

3. 20 mg Remdesivir nucleoside monophosphate (Reference standard for the assay development. Since compound is not stable in solution, extra material is needed for each assay run)

4. 50 mg GS-443902 (triphosphate GS-441524) (Reference standard for the assay development. Since compound is not stable in solution, extra material is needed for each assay run)

LEVEL OF EFFORT:

N/A

GOVERNMENT RESPONSIBILITIES

NCATS will provide the Contractor with ~ 10 mg each of Remdesivir and GS-441524.

DELIVERY OR DELIVERABLES

The CRO lab shall deliver the following by email to xin.xu3@nih.gov:

1. Quantitative and sensitive assays for Remdesivir, GS-441524, GS-441524 mono- and tri-phosphates

2. Method qualification to evaluate the linearity, precision, accuracy, recovery, and carry over of the assays, and the stability of analytes during assay process.

3. Quantification of Remdesivir, GS-441524, GS-441524 mono- and tri-phosphates in lysates collected from in vitro incubations with three cell lines (Vera E6, Calu3, HEK293).

REPORTING REQUIREMENTS

The CRO lab shall deliver the reports on bioanalytical method development and concentration values for Remdesivir, GS-441524, GS-441524 mono- and tri-phosphates in lysates by email to xin.xu3@nih.gov.

OTHER CONSIDERATIONS

Travel:

N/A

Key Personnel:

Scientists with solid experience and scientific knowledge on bioanalytical method development for nucleoside analogs and the polar phosphate metabolites.

Information System Security Plan:

Standard CRO lab Information System Security Plan.

Data Rights:

The Government retains unlimited rights in all data generated under this contract.

Section 508—Electronic and Information Technology Standards:

SOP of CRO lab

Publications and Publicity:

There shall be no publication of the data, results, findings, or conclusions generated under this contract without prior approval from the Government.

Confidentiality of Information:

The contractor, including all its employees, consultants, and subcontractors, shall maintain confidentiality of all information generated under this project.

mailto:xin.xu3@nih.gov mailto:xin.xu3@nih.gov