1.5369134_SOW.pdf

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

STATEMENT OF WORK (SERVICES)

(SOW)

GENERAL INFORMATION

Title of Project:

1. ADME services (Solubility, PAMPA, and Microsomal Stability)

2. ADME services (Express LC-MS/MS Method Development, Hepatocyte Stability (Mouse + Rat))

3. ADME services (Express LC-MS/MS Method Development, Caco-2 Permeability (Bi-directional))

Statement of Need and Purpose:

1. The purpose of this acquisition is to obtain high quality, multiple replicate, aqueous solubility, PAMPA permeability and microsomal stability assessments for a series of ten (10) compounds.

2. This order provides for testing of twenty (20) small molecule compounds for stability in mouse and rat hepatocytes. These compounds are drug molecules being evaluated for the dual IRAK4-FLT3 kinase program. Testing of the compounds for their stability in mouse and rat hepatocytes will provide critical information on their clearance properties, and will inform subsequent in vivo pharmacokinetic studies in mouse and rat.

3. This order provides for testing of twenty (20) small molecule compounds for permeability in Caco-2 cells.

These compounds are drug molecules being evaluated for the dual IRAK4-FLT3 kinase program. Testing of the compounds for permeability and active transport in Caco-2 cells will provide critical information on their transport properties, and will allow us to prioritize the development of compounds with optimal transport properties.

Background Information and Objective:

The National Institutes of Health (NIH) is the nation’s leading medical research agency and the primary Federal agency whose mission is to seek fundamental knowledge about the nature and behavior of living systems and the application of that knowledge to enhance health, lengthen life, and reduce illness and disability, conducting, supporting and making medical discoveries that improve people’s health and save lives. The National Center for Advancing Translational Sciences (NCATS) is a translational science center that prides itself on its tremendously productive pipeline and is innovative in a number of ways. NCATS brings together a diverse range of scientists, including medicinal chemists, biologists, toxicologists, and engineers, in order to ultimately translate basic science into real products and services that help improve people’s lives.

(1) NCATS is currently interested select compound assessments, specifically aqueous solubility, PAMPA permeability, and rat microsomal stability. Due to the nature of these measurements, it is required they have high precision and accuracy.

(2) Prior work at NCATS has produced a series of small molecule IRAK4 / FLT3 kinase dual inhibitors, including NCGC00371481 and its structural analogs. Compounds such as ‘1481 have shown promising results in cellular and mouse models of myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML).

Further optimization of the biochemical, physiochemical and pharmacokinetic properties of these compounds is required, and may eventually produce a clinical candidate that could be used to treat cancers such as MDS and / or AML.

In order for a candidate compound to advance into clinical trials, it must display a wide range of acceptable properties. For this program, a potential clinical candidate must display potent inhibition of the desired targets (IRAK4 and FLT3). It must also display suitable pharmacokinetic properties in a range of preclinical species, including at least one rodent species (mouse or rat), as well as at least one higher preclinical species (dog or monkey). Good pharmacokinetic profiles in these species are required 1) in order to build models that predict good pharmacokinetic properties in humans, and 2) so that compounds can be given at high doses and high plasma exposure levels in preclinical safety studies.

Of the pharmacokinetic properties that must be optimized, drug clearance is perhaps the most important one. Clearance can be measured in vivo, in pharmacokinetic studies run in a preclinical species (mouse, rat, dog, or monkey), but such studies are expensive. A more cost-effective way to measure clearance involves in vitro studies, by incubating the test compound with either liver microsomes or hepatocytes. Of those two, incubation with hepatocytes is preferred, as it is typically more predictive of in vivo outcomes.

For a given compound, clearance in hepatocytes may vary considerably across species. That is, clearance in mouse, rat, dog, monkey or human hepatocytes may vary. For that reason, preclinical drug discovery programs that run experiments in a variety of preclinical species, and that want to advance compounds into human clinical trials, will ultimately need to test for clearance in mouse, rat, dog, monkey and human hepatocytes.

Testing of a set of 20 small molecule compounds for clearance in mouse and rat hepatocytes will allow a determination of the compounds’ clearance rates. This will allow us to identify compounds with optimal clearance rates, and to preferentially select those for subsequent, more expensive pharmacokinetic studies in mouse and rat.

(3) Prior work at NCATS has produced a series of small molecule IRAK4 / FLT3 kinase dual inhibitors, including NCGC00371481 and its structural analogs. Compounds such as ‘1481 have shown promising results in cellular and mouse models of myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML).

Further optimization of the biochemical, physiochemical and pharmacokinetic properties of these compounds is required, and may eventually produce a clinical candidate that could be used to treat cancers such as MDS and / or AML.

In order for a candidate compound to advance into clinical trials, it must display a wide range of acceptable properties. For this program, a potential clinical candidate must display potent inhibition of the desired targets (IRAK4 and FLT3). It must also display good cell permeability, so that it can effectively absorbed across cell membranes into systemic circulation, and ultimately be delivered to the target tissue of interest.

Caco-2 cells are widely used as a model cell line for determining a compound’s cell permeability. In these experiments, a test compound can be applied to either the apical (A) or basolateral (B) side of the cell membrane. If a compound is applied to the apical (A) side, one then takes samples from the basolateral (B) side to see how well the compound permeates the cell membrane. The experiment can also be run in reverse, wherein compound is applied first to the basolateral (B) side, with samples then being taken from the apical (A) side. If the experiment is run in two parts – first applying compound to side A and sampling from side B, and then vice versa – one can determine if a compound is being preferentially pumped into or out of the cell. This bidirectional Caco-2 cell experiment provides information not only on passive cell permeability, but also on whether the test compound is a substrate for active transport into or out of the cell (uptake or efflux transport).

Testing of a set of 20 small molecule compounds for permeability in bidirectional Caco-2 cell assays will allow a determination of the compounds’ passive cell permeability, as well as a determination of whether they are substrates for active uptake or efflux transporters. This will allow us to identify compounds with optimal transport properties, and to preferentially select those for subsequent, more advanced experiments.

Period of Performance:

1. Period of performance is 3 months from the award date or compound receipt.

2. Period of performance is within 2 months from the contract award and receipt of first batch of material

3. Period of performance is within 2 months from the contract award and receipt of first batch of material

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:

1. ADME Services (Solubility, PAMPA, and Microsomal Stability)

a. Ten different compounds will be assessed in each of these assays (Solubility, PAMPA and

Metabolic stability)

b. Contractor will perform aqueous kinetic solubility measurements on the target compounds

c. Contractor will provide PAMPA permeability assessment for the target compounds

d. Contractor will provide a metabolic stability assessment for the target compounds, utilizing rat liver microsomes

e. Contractor will present a report with the above results.

2. ADME services (Express LC-MS/MS Method Development, Hepatocyte Stability (Mouse + Rat))

1. Test each of 20 small molecule compounds for clearance in mouse and rat hepatocytes.

a. Provide a comprehensive report with the results of the experiments of each of the 20 compounds tested for clearance in mouse and rat hepatocytes.

3. ADME services (Express LC-MS/MS Method Development, Caco-2 Permeability (Bi-directional))

1. Test each of 20 small molecule compounds for permeability and active transport in bidirectional Caco-2 cell assays.

a. Provide a comprehensive report with the results of the experiments of each of the 20 compounds tested for permeability and active transport in bidirectional Caco-2 cell assays.

Specific Requirements:

Technical Requirements:

a. Ten different compounds will be assessed in each of these assays (Solubility, PAMPA and

Metabolic stability)

b. Contractor will perform aqueous kinetic solubility measurements on the target compounds

i. Compounds will be provided as a solution in DMSO

ii. 7 replicates will be used.

iii. A range of final concentrations for the target compound will be used

iv. A range of <0.5 ug/ml to >20 ug/ml must be able to be determined from this assay

v. A high solubility and low solubility control of the contractor’s choice will also be included.

c. Contractor will provide PAMPA permeability assessment for the target compounds

i. Four replicates for each compound will be determined

ii. A range of 0.1 to >60 10-6 cm/s must be able to be determined by the assay

iii. A high permeability control and low permeability control of the contractor’s choice will also be included.

d. Contractor will provide a metabolic stability assessment for the target compounds, utilizing rat liver microsomes

i. Four different non-zero time points and a zero time point will be used for each compound.

ii. Two different control compounds, of the contractor’s choice, will also be utilized

iii. Percent compound remaining will be determined and reported at each time point

iv. Intrinsic Clearance will be calculated and reported for each compound

v. T1/2 will be calculated and reported for each compound

e. Contractor will present a report with the above results.

2. ADME Services (Express LC-MS/MS Method Development, Hepatocyte Stability (Mouse + Rat))

a. Contractor will run the assays using scientists that have significant experience (>5 years) in the testing of small molecule compounds for clearance in mouse and rat hepatocytes.

b. Contractor will have significant experience (>5 years) in running assays that test for binding to mouse and human plasma protein.

3. ADME Services (Express LC-MS/MS Method Development, Caco-2 Permeability (Bi-directional))

a. Contractor will run the assays using scientists that have significant experience (>5 years) in the testing of small molecule compounds for permeability and active transport in bidirectional Caco-2 cell assays.

b. Contractor will have significant experience (>5 years) in running assays that test for permeability and active transport in bidirectional Caco-2 cell assays.

LEVEL OF EFFORT:

- One Ph.D. level manager scientist, 2 hours

- Three Bachelor’s level biochemists, 4 hours

2. ADME Services (Express LC-MS/MS Method Development, Hepatocyte Stability (Mouse + Rat))

- Estimated personnel: 1 PhD level-Manager. 1 Ph.D. level scientist. 1-2 months total.

3. ADME Services (Express LC-MS/MS Method Development, Caco-2 Permeability (Bi-directional))

- Estimated personnel: 1 PhD level-Manager. 1 Ph.D. level scientist. 1-2 months total.

GOVERNMENT RESPONSIBILITIES

Government has the responsibility to review the data, reports, and products generated. Government’s responsibility includes final approval on reports

Government will not provide telephones, workspace, or desktop computers. Contractor will not require access to government facilities.

Government will not furnish property, facilities, workspace, computers, or other equipment. Contractor will not use federal facilities. Government will have the responsibility to review and approve the final report. Government will have the responsibility to provide sufficient quantities of the lead compound in order to contractor to accomplish the necessary work

DELIVERY OR DELIVERABLES

a. One report for the given data, as requested

2. ADME Services (Express LC-MS/MS Method Development, Hepatocyte Stability (Mouse + Rat))

a. Contractor will provide the data analysis and report for the contents of this statement of work.

3. ADME Services (Express LC-MS/MS Method Development, Caco-2 Permeability (Bi-directional))

a. Contractor will provide the data analysis and report for the contents of this statement of work.

REPORTING REQUIREMENTS

1. Contractor will be expected to stay in e-mail contact with the government representative, as needed

2. Contractor will provide monthly updates on the progress of the project, or more often as appropriate

OTHER CONSIDERATIONS

Travel:

N/A

Key Personnel:

Information System Security Plan:

Data Rights:

Data may be produced as part of this contract. The government retains rights to this data.

Section 508—Electronic and Information Technology Standards:

Contractor is not required to develop, procure, maintain or use EIT

Publications and Publicity:

Contractor will not write, nor publish articles, except with the express written permission of the government.

Confidentiality of Information:

Contractor will keep data generated strictly confidential. Any information the contractor is to release must be authorized by the government before release.