D1 SOW 011222.pdf

STATEMENT OF WORK

(SOW)

GENERAL INFORMATION

Title of Project:

Protein Crystallization and X-Ray Crystallography on Large tumor suppressor kinase 1 (LATS1)

Statement of Need and Purpose:

The Division of Preclinical Innovation (DPI) within the National Center for Advancing

Translational Sciences (NCATS) requires three (3) x-ray structures of the LATS1, both alone and in complex with two (2) small molecule inhibitor, in order to further the current development of small molecule inhibitors of LATS1.

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.

NCATS is one of 27 Institutes and Centers at the NIH and its mission is to catalyze the generation of innovative methods and technologies that will enhance the development, testing and implementation of diagnostics and therapeutics across a wide range of human diseases and conditions. The NCATS DPI plans, conducts and uses both internal and contract resources to advance collaborative research projects across the preclinical phases of the translational science spectrum.

The kinase LATS1 is of interest to NCATS in several different disease indications. Currently, there is no x-ray structure of LATS1. In order to further the current development of small molecule inhibitors of LATS1, a high-quality x-ray structure of the protein, both alone and in complex with small molecule inhibitors is required by NCATS.

The X-ray structure will provide in depth structural information about the biophysical conformation of the protein. This structural information can be used to better design small molecule inhibitors of the protein.

Period of Performance:

The anticipated period of performance is January 21, 2022 through January 20, 2023 (12-months).

SCOPE OF WORK

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 specific requirements of this Statement of Work.

Specific Requirements:

The Contractor shall perform the services in four (4) phases as follows:

Phase 1. CONSTRUCT DESIGN

• The contractor shall design five (5) different constructs for large tumor suppressor kinase

1 (LATS1) and at least one (1) construct for any LATS1-associated proteins and express them in E. coli for suitable expression levels evaluation.

o Even though some of the constructs may not be the full length LATS1 protein, they must all contain the LATS1 kinase domain (IE, the ATP pocket).

• A report on the results on the E.Coli expression shall be provided to NCATS for analysis and determination of constructs for phase 2.

o In case the constructs are not acceptable by NCATS, the constructs shall be revised and approved before moving to phase 2.

Phase 2. PROTEIN EXPRESSION/PURIFICATION

• NCATS shall determine one (1) construct of LATS1 to be expressed in E. coli in sufficient quantities for purification and crystallization studies.

• NCATS shall determine one (1) construct of LATS1 in combination with the LATS1-associated proteins to be expressed in E. coli in sufficient quantities for purification and crystallization studies.

• The resulting proteins shall be purified via standard chromatography methods and shall be characterized by standard methods, including but not limited to SDS-Page and Mass spectrometry.

• A report on the results of phase 2 shall be provided to NCATS for analysis and approval for phase 3.

Phase 3. CRYSTALLIZATION AND CO-CRYSTALLIZATION SCREENING

• Each of the protein constructs from phase 2 shall be screened for conditions to provide

X-ray quality crystals.

• Each of the protein constructs from phase 2 shall be screened in complexation with two

(2) small molecule inhibitors of LATS1 (to be provided by NCATS) for conditions to provide X-ray quality crystals.

• Optimal conditions shall be identified, optimized, and scaled up in order to provide enough material for X-ray crystallographic analysis.

• A report on the results of phase 3 shall be provided to NCATS for analysis and approval for phase 4.

Phase 4. SYNCHOTRON X-RAY AND X-RAY STRUCTURE DETERMINATION

• Suitable crystals shall be examined by X-ray diffraction with desired resolution of less than three (3) Angstroms.

• If the diffraction data provides sufficient quality, the full X-ray data set for the crystals shall be obtained.

• The data sets shall be indexed, processed, and scaled within common standards to the field.

• The data sets shall be refined and examined to provide a model, assuming the data and electron density maps are of sufficient quality.

• The model shall be presented to NCATS. The structure files shall be provided as needed at least on the following formats: .pdb, .mtz, and .cif.

• The model shall include the following:

o One (1) X-ray crystallographic structure containing the LATS1 protein without small molecule inhibitor;

o One (1) X-ray crystallographic structure model containing the LATS1 protein in complex with NCATS small molecule inhibitor one (1); and o One (1) X-ray crystallographic structure model containing the LATS1 protein in complex with NCATS small molecule inhibitor two (2).

i. In case the three (3) desired crystallographic structures are deemed not possible to be obtained by NCATS and the Contractor, the contractor shall provide the model, if any, with the achieved crystallographic structure(s) and a report with the results of phase 4.

Government Responsibilities:

The Government will not furnish property, facilities, workspace, computers, or other equipment.

Performance of the services shall be on-site at the Contractor facility. The Government will have the responsibility to review and approve the reports and provide two (2) small molecule inhibitors of LATS1 in order for the contractor to perform the services described herein the

SOW.

Delivery or Deliverables:

The contractor shall provide interim reports at the conclusion of each phase for review and approval by NCATS before moving to the next phase. The interim reports shall be sent electronically to [to be specified at time of award].

The contractor shall provide one (1) final report encompassing the results of all phases of the specific requirements of this Statement of Work as well as the models obtained as a result of the services specified on phase 4. The final report shall be sent electronically to [to be specified at time of award].

In addition to the final report, the contractor shall arrange a virtual meeting with [to be specified at time of award] and other members of NCATS in order to present the results of the services.

Reporting Requirements:

The Contractor shall provide by-weekly updates on the progress of the project, or more often as appropriate. Updates shall be provided electronically to [to be specified at time of award].

Other Considerations:

Data Rights:

FAR Clause 52.227-14, Rights in Data-General (May 2014) is included in the solicitation and resultant award. Under 52.227-14(c)(1), the contractor must obtain approval from the contracting officer to assert copyright in any data first produced in performance of this contract and published in academic, technical or professional journals, symposia proceedings, or similar works. Under 52.227-14(d)(2), the contractor must obtain approval from the contracting officer to use, release to others, reproduce, distribute, or publish any data first produced or specifically used by the contractor in the performance of this contract.

In addition, the Contractor shall keep the results of the acquisition confidential.