Through this Notice of Funding Opportunity (NOFO), the National Cancer Institute (NCI) solicits applications for identification of small molecules that function to elucidate the biology of disease as chemical probes or function as agonists or antagonists of disease target(s) for therapy or immunotherapy. The NOFO is intended to support discovery research for the identification of validated hits relevant to health-related outcomes of participating NIH Institutes. Stages of discovery research covered by this NOFO include: 1) assay development for specific biological targets and disease mechanisms relevant to the mission of participating NIH Institutes with the intent to screen for small molecule compounds that show potential as probes for use in advancing knowledge about the known targets, identifying new targets, or as pre-therapeutic leads; 2) screen implementation high throughput target-focused approaches or moderate throughput phenotypic- and fragment-based approaches to identify initial screening hits; 3) hit validation, including implementation of secondary assays that are orthogonal to the primary assay, advanced cheminformatics analysis and initial medicinal chemistry inspection to prioritize the hit set, and follow-up assays to characterize mode and mechanism of action of the validated hits; 4) hit-to-lead optimization, including SAR to optimize target engagement, selectivity and to minimize chemical liabilities, ADME, PK and PD studies, and, if appropriate, in vivo modeling to test efficacy or biological effects.

PAR-25-153: Assay development and screening for discovery of chemical probes, drugs or immunomodulators (R01 Clinical Trial Not Allowed)

Posted Date: November 06, 2024 Open Date (Earliest Submission Date): January 05, 2025 Expiration Date: September 08, 2026

This funding opportunity was updated to align with agency priorities. Carefully reread the full funding opportunity and make any needed adjustments to your application prior to submission.

Part 1. Overview Information

Participating Organization(s): National Institutes of Health (NIH) Components of Participating Organizations: National Cancer Institute (NCI)

Funding Opportunity Title: Assay development and screening for discovery of chemical probes, drugs or immunomodulators (R01 Clinical Trial Not Allowed) Activity Code: R01 Research Project Grant Announcement Type: Reissue of PAR-23-264

Related Notices:

• March 31, 2025: This funding opportunity was updated to align with agency priorities. Carefully reread the full funding opportunity and make any needed adjustments to your application prior to submission.
• January 7, 2025: Notice of Correction to PAR-25-153 Assay development and screening for discovery of chemical probes, drugs or immunomodulators (R01 Clinical Trial Not Allowed). See Notice NOT-CA-25-030.
• April 4, 2024: Overview of Grant Application and Review Changes for Due Dates on or after January 25, 2025. See Notice NOT-OD-24-084.
• August 31, 2022: Implementation Changes for Genomic Data Sharing Plans Included with Applications Due on or after January 25, 2023. See Notice NOT-OD-22-198.
• August 5, 2022: Implementation Details for the NIH Data Management and Sharing Policy. See Notice NOT-OD-22-189.

Funding Opportunity Number (FON): PAR-25-153 Companion Funding Opportunity: None Number of Applications: See Section III. 3. Additional Information on Eligibility Assistance Listing Number(s): 93.395

Funding Opportunity Purpose: Through this Notice of Funding Opportunity (NOFO), the National Cancer Institute (NCI) intends to stimulate research in discovery and development of novel, small molecules for cancer. Molecules discovered through this NOFO may be used to probe cancer biology, to validate cancer targets, or as the basis for optimized drugs.

Stages of discovery research covered by this NOFO include:

1. Development of the primary screen assay(s) and testing in an initial pilot screen.
2. Primary screen implementation to identify initial screening hits (high throughput target-focused screens, or moderate throughput screens).
3. Hit validation using a series of assays and initial medicinal chemistry inspection to prioritize the hit set.

Key Dates:

All applications are due by 5:00 PM local time of applicant organization. Applicants are encouraged to apply early to allow adequate time to make any corrections to errors found in the application during the submission process by the due date.

Due Dates for E.O. 12372: Not Applicable

Required Application Instructions: It is critical that applicants follow the instructions in the Research (R) Instructions in the How to Apply - Application Guide, except where instructed to do otherwise (in this NOFO or in a Notice from NIH Guide for Grants and Contracts). Conformance to all requirements (both in the Application Guide and the NOFO) is required and strictly enforced. Applicants must read and follow all application instructions in the Application Guide as well as any program-specific instructions noted in Section IV. When the program-specific instructions deviate from those in the Application Guide, follow the program-specific instructions. Applications that do not comply with these instructions may be delayed or not accepted for review.

There are several options available to submit your application through Grants.gov to NIH and Department of Health and Human Services partners. You must use one of these submission options to access the application forms for this opportunity.

• Use the NIH ASSIST system to prepare, submit and track your application online.
• Use an institutional system-to-system (S2S) solution to prepare and submit your application to Grants.gov and eRA Commons to track your application. Check with your institutional officials regarding availability.
• Use Grants.gov Workspace to prepare and submit your application and eRA Commons to track your application.

Part 2. Full Text of Announcement

Section I. Notice of Funding Opportunity Description

Purpose & Background

Through this Notice of Funding Opportunity (NOFO), the National Cancer Institute (NCI) intends to support discovery research for the identification of small molecules that function to elucidate the biology of cancer as chemical probes or function as activators or inhibitors of cancer target(s) for therapy. The scientific rationale for target selection must be provided. Small molecule tools or therapeutics may target tumor cells themselves or immune cells regulating tumor growth.

Stages of discovery research covered by this NOFO include:

1. Development of the primary screen assay(s) and testing in an initial pilot screen. Assays may focus on specific biological targets or disease mechanisms relevant to cancer with the intent to screen for small molecule compounds to be used as probes for advancing knowledge about the known target(s), identifying new targets, or as pre-therapeutic leads.
2. Screen implementation of high throughput target-focused approaches or moderate throughput phenotypic- and fragment-based approaches to identify initial screening hits.
3. Hit validation, including implementation of secondary assays that are orthogonal to the primary assay, advanced cheminformatics analysis and initial medicinal chemistry inspection to prioritize the hit set, and follow-up assays to characterize mode and mechanism of action of the validated hits.

It is anticipated that applications submitted in response to this NOFO will focus on one or two of the above stages, as each stage depends on success of the prior stage.

The small molecules for targets to be pursued must be relevant to understanding cancer biology, and/or for developing strategies for cancer prevention, diagnosis, treatment or clinical monitoring of treatment. In addition to targeting tumor cells themselves, applications targeting immune cells that regulate tumor development are appropriate for this NOFO. Small molecules that affect known immune checkpoint inhibitor pathways as well as small molecules that address emerging pathways involved in immune-mediated control of cancer are of interest. NCI also encourages projects focusing on small molecules that target pediatric fusion oncoproteins or that address novel targets in small cell lung cancer or pancreatic cancer. Applicants may find the NCI Developmental Therapeutics Program resources to be helpful. These include the NCI Program for Natural Product Discovery (NPNPD) Prefractionated Library which, as of March 2023, consisted of 500,000 natural product fractions that are available for screening.

Assessment of the reasons for failures of small molecule therapeutic agents in the clinic points to several areas for improvement of the drug discovery and development process that are pertinent to this NOFO:

• First, increased rigor in target identification is necessary. For instance, whether prior studies of the selected target were adequately controlled and powered are important considerations. Were cell lines verified, plasmids sequenced, and protein reagents tested for contaminants?
• Second, reproducibility of the proposed primary assay should be carefully considered because this assay is often the basis for assessing not only initial hits but also for iteratively assessing optimized hits during structure-activity relationship (SAR) studies. Development of primary screening assays that test a key biological function of the target of interest are likely to yield hits of increased relevance. In this respect, phenotypic screens have had a resurgence.
• Third, a hit validation scheme or “critical path” that includes orthogonal assay(s) to eliminate false positives, as well as a series of assays in diverse biological systems with diverse read-outs, particularly including assays that model human disease, is likely to yield hits of increased relevance.
• Fourth, the inclusion of skilled synthetic and/or medicinal chemists to assess the validity of the hit chemotypes to eliminate PAINS (pan-assay interference compounds) or other undesirable chemotypes is likely to be beneficial.

Although outside the scope of this NOFO, extensive chemical optimization of hit molecules to generate highly selective probes or drugs is a logical next step. Databases of small-molecule probes for the study of disease-related biological processes and mechanisms in academic environments may be found in the Academic Drug Discovery Consortium or the Chemical Probes Portal. It is noted that probes may be the predecessors of drugs, but drugs with known off-target effects are seldom useful as probes of specific biological activities. Importantly, identification of chemical probe(s) for a given target opens the door to validation of the target in appropriate cellular, tissue, or animal models prior to clinical testing.

Research Scope

Projects for this NOFO may focus on one stage, or span several stages of discovery research:

1. Development of the primary screen assay(s) and testing in an initial pilot screen
2. Primary screen implementation
3. Hit validation

For applications requesting support for more than one stage, demonstration of feasibility is needed, including strong justification and supporting preliminary data for the stages proposed.

Areas to be considered for each stage are described below and given in greater detail in Section IV (Application and Submission Information, PHS 398 Research Plan).

1. Development of primary screen assay(s) and testing in an initial pilot screen

Through this NOFO, NCI seeks to apply new knowledge and screening technologies to develop assays for novel cancer targets and pathways. Projects for assay development should emphasize the design and validation of creative approaches that have the potential to be used for chemical probe or drug discovery. Assays focusing on areas and approaches that have been extensively studied should be avoided unless a strong rationale is provided for additional studies in the projected area. Targets associated with rare and neglected cancers are encouraged.

Primary screening assays may be target-, pathway-, or phenotype-based. Some examples include:

• Target-based biochemical or cellular assays that measure activities of enzymes, receptor-ligand binding, protein-protein interactions, ion channels, transporters, nuclear receptors, and other transcription factors, and other new targets emerging from genetic and proteomic research in model systems and in human cancers.
• Cell- or organism-based assays that detect phenotypic changes that may involve unidentified molecular targets.
• Non-traditional targets of interest such as nucleic acids, protein folding, polymorphic gene products, post-transcriptional editing or gene-splicing factors, and protein or RNA stabilizers.

Assay detection methods may include, but are not limited to:

• Fluorescence, luminescence, absorbance
• Fluorescence resonance energy transfer (FRET)
• Time-resolved fluorescence resonance energy transfer (TR-FRET)
• Fluorescence polarization
• Flow cytometric measurements
• Fluorescence imaging
• Bioluminescence resonance energy transfer (BRET)
• AlphaScreen, scintillation proximity assay (SPA)
• Electrophysiological assays
• Biophysical assays

In general, assays should adopt an adequate detection principle that results in the sensitive detection of even weak binders with expected low rates of false positives and false negatives. Complementary research including virtual screening may also be conducted to improve the likelihood of success and cost-effectiveness.

If achievable, pilot screening for target validation from a small compound library (e.g., LOPAC1280), should be proposed as a highly desirable validation step for translation of the assay to HTS. Instructions for the design and testing of a primary assay may be found in Section IV (Application and Submission Information, PHS 398 Research Plan).

2. Primary Screen Implementation

Projects focusing on screen implementation are encouraged to provide preliminary data demonstrating that a primary screen has been developed, fully characterized, and tested in a pilot format. See Section IV for further details on expected information prior to the Primary Screen Implementation stage.

Virtual Screening (VS)

In addition to screens of chemical libraries, NCI, through this NOFO, seeks to make use of large, publicly available databases of 3D protein structures, genomic metabolomic, and proteomic data, and virtual small molecule libraries, combined with computational approaches to accelerate the discovery of novel and potential drug candidates based on specific molecular targets. Projects proposing the use of VS, either ligand-based screening or receptor-based screening, must collaborate with hit assay validation and medicinal chemistry experts for experimental testing of the proposed in silico hits.

VS techniques may involve computational approaches that include, but are not limited to:

• Machine learning based QSAR (particularly deep learning methods)
• Innovative protein-ligand docking algorithms
• Similarity search methods
• Pharmacophore mapping methods

Applicants are encouraged to collaborate with an experienced screening facility, particularly if high throughput screening (HTS) is planned. The screening facility may provide advice such as identification and selection of commercial HTS assay reagents, and suitable HTS assay format and readout. In addition, the screening facility may be able to aid in adapting assays to an HTS format (e.g., 1536-well or 384-well microplate). Further, the researchers might seek advice from the screening laboratory about orthogonal assays to validate the screening hits. Phenotypic screening should only be proposed if target deconvolution is included in the application or if there is a strong association between the mechanisms that drive the phenotypic assay, the preclinical disease model, and the human disease.

Technical resources about assay development and screening include the online comprehensive guidebook (Assay Guidance Manual), assay protocols deposited on the PubChem BioAssay data base, ASSAY and Drug Development Technologies, a peer-reviewed bi-monthly journal, and SLAS Discovery.

3. Hit validation

For the purposes of this NOFO, a “hit” is defined as a compound that has the desired activity in a compound screen and whose activity is confirmed upon retesting in orthogonal assays.

Hits from a primary screen may be systematically assessed using a cascade of follow-up assays to remove false positives. Primary HTS assays typically generate hundreds to thousands of hits, many of which are false positives or are chemically intractable. Hits from smaller scale primary screens are also likely to generate false positives or chemically intractable molecules that require additional screens. Post-primary screening assays may include:

• An assay that is essentially identical to the primary assay but with an orthogonal detection scheme (e.g., switching light detection mode or wavelength to avoid intrinsic compound interference).
• A target-minus assay (e.g., coupling enzymes in the absence of the assay target enzyme, parental cell line without the assay target protein, etc.).
• An assay that is different in biological context and process (e.g., protein functional assay vs. protein binding assay; RT-PCR and Western assay vs. reporter gene assay; cell-based assay vs. biochemical assay).
• Cytotoxicity assay.
• Target selectivity assay(s).
• Specificity assays to distinguish biological activities of chemical entities among orthologous targets across organism species through kingdoms (e.g., yeast vs. mammalian cell targets).
• Mode of action assays (e.g., allosteric vs. orthosteric; competitive vs. non-competitive or uncompetitive).
• Target identification assays.

The assays farther downstream may also include cellular and tissue models pertaining to the relevant physiology or pathophysiology, or to mode and mechanism of action of the validated hits. In vivo assays (e.g., whole animal models) should only be proposed if they are needed to demonstrate the biological or physiological effects of lead compound(s).

In addition, investigators should conduct advanced cheminformatics analysis and medicinal chemistry inspection to prioritize the hit set. It is expected that the investigators will test powder samples of hit compounds and commercially available analog compounds during the hit validation stage. Investigators should verify the structure of hits using a combination of analytical methods and, if possible, re-synthesis of select hits.

Nonresponsive Applications

The following types of studies are beyond the scope of this NOFO. Applications proposing such studies will be considered non-responsive and will not be reviewed.

• Targets and associated screens and assays that are not relevant to cancer.
• Detailed structure-activity relationship (SAR) studies.
• In vitro or in vivo Absorption, Distribution, Metabolism, and Excretion (ADME) assays.
• In vivo animal assays, unless justified as necessary for screening of hit molecule(s).

See Section VIII. Other Information for award authorities and regulations.

Section II. Award Information

Funding Instrument: Grant: A financial assistance mechanism providing money, property, or both to an eligible entity to carry out an approved project or activity.

Application Types Allowed:

• New
• Renewal
• Resubmission
• Revision

The OER Glossary and the How to Apply Application Guide provide details on these application types. Only those application types listed here are allowed for this NOFO.

Clinical Trial? Not Allowed: Only accepting applications that do not propose clinical trials. Need help determining whether you are doing a clinical trial?

Funds Available and Anticipated Number of Awards: The number of awards is contingent upon NIH appropriations and the submission of a sufficient number of meritorious applications.

Award Budget: Application budgets are not limited but need to reflect the actual needs of the proposed project.

Award Project Period: The total project period may not exceed 3 years.

NIH grants policies as described in the NIH Grants Policy Statement will apply to the applications submitted and awards made from this NOFO.

Section III. Eligibility Information

1. Eligible Applicants

Eligible Organizations:

• Higher Education Institutions
  • Public/State Controlled Institutions of Higher Education
  • Private Institutions of Higher Education
• Nonprofits Other Than Institutions of Higher Education
  • Nonprofits with 501(c)(3) IRS Status (Other than Institutions of Higher Education)
  • Nonprofits without 501(c)(3) IRS Status (Other than Institutions of Higher Education)
• For-Profit Organizations
  • Small Businesses
  • For-Profit Organizations (Other than Small Businesses)
• Local Governments
• State Governments
• County Governments
• City or Township Governments
• Special District Governments
• Indian/Native American Tribal Governments (Federally Recognized)
• Indian/Native American Tribal Governments (Other than Federally Recognized)
• Federal Governments
  • Eligible Agencies of the Federal Government
• U.S. Territory or Possession
• Other
  • Independent School Districts
  • Public Housing Authorities/Indian Housing Authorities
  • Native American Tribal Organizations (other than Federally recognized tribal governments)
  • Faith-based or Community-based Organizations
  • Regional Organizations
• Non-domestic (non-U.S.) Entities (Foreign Organizations)
  • Foreign Organizations
  • Non-domestic (non-U.S.) Entities (Foreign Organizations) are eligible to apply.
  • Non-domestic (non-U.S.) components of U.S. Organizations are eligible to apply.
  • Foreign components, as defined in the NIH Grants Policy Statement, are allowed.

Required Registrations:

Applicant Organizations: Applicant organizations must complete and maintain the following registrations as described in the How to Apply- Application Guide to be eligible to apply for or receive an award. All registrations must be completed prior to the application being submitted. Registration can take 6 weeks or more, so applicants should begin the registration process as soon as possible. Failure to complete registrations in advance of a due date is not a valid reason for a late submission, please reference the NIH Grants Policy Statement Section 2.3.9.2 Electronically Submitted Applications for additional information.

• System for Award Management (SAM) – Applicants must complete and maintain an active registration, which requires renewal at least annually. The renewal process may require as much time as the initial registration. SAM registration includes the assignment of a Commercial and Government Entity (CAGE) Code for domestic organizations which have not already been assigned a CAGE Code.
• NATO Commercial and Government Entity (NCAGE) Code – Foreign organizations must obtain an NCAGE code (in lieu of a CAGE code) in order to register in SAM.
• Unique Entity Identifier (UEI) - A UEI is issued as part of the SAM.gov registration process. The same UEI must be used for all registrations, as well as on the grant application.
• eRA Commons - Once the unique organization identifier is established, organizations can register with eRA Commons in tandem with completing their Grants.gov registrations; all registrations must be in place by time of submission. eRA Commons requires organizations to identify at least one Signing Official (SO) and at least one Program Director/Principal Investigator (PD/PI) account in order to submit an application.
• Grants.gov – Applicants must have an active SAM registration in order to complete the Grants.gov registration.

Program Directors/Principal Investigators (PD(s)/PI(s)): All PD(s)/PI(s) must have an eRA Commons account. PD(s)/PI(s) should work with their organizational officials to either create a new account or to affiliate their existing account with the applicant organization in eRA Commons. If the PD/PI is also the organizational Signing Official, they must have two distinct eRA Commons accounts, one for each role. Obtaining an eRA Commons account can take up to 2 weeks.

Eligible Individuals (Program Director/Principal Investigator): Any individual(s) with the skills, knowledge, and resources necessary to carry out the proposed research as the Program Director(s)/Principal Investigator(s) (PD(s)/PI(s)) is invited to work with their organization to develop an application for support. For institutions/organizations proposing multiple PDs/PIs, visit the Multiple Program Director/Principal Investigator Policy and submission details in the Senior/Key Person Profile (Expanded) Component of the How to Apply-Application Guide.

2. Cost Sharing: This NOFO does not require cost sharing as defined in the NIH Grants Policy Statement Section 1.2 Definition of Terms.

3. Additional Information on Eligibility

Number of Applications: Applicant organizations may submit more than one application, provided that each application is scientifically distinct. The NIH will not accept duplicate or highly overlapping applications under review at the same time, per NIH Grants Policy Statement Section 2.3.7.4 Submission of Resubmission Application. This means that the NIH will not accept:

• A new (A0) application that is submitted before issuance of the summary statement from the review of an overlapping new (A0) or resubmission (A1) application.
• A resubmission (A1) application that is submitted before issuance of the summary statement from the review of the previous new (A0) application.
• An application that has substantial overlap with another application pending appeal of initial peer review (see NIH Grants Policy Statement 2.3.9.4 Similar, Essentially Identical, or Identical Applications).

Section IV. Application and Submission Information

1. Requesting an Application Package: The application forms package specific to this opportunity must be accessed through ASSIST, Grants.gov Workspace or an institutional system-to-system solution. Links to apply using ASSIST or Grants.gov Workspace are available in Part 1 of this NOFO. See your administrative office for instructions if you plan to use an institutional system-to-system solution.

2. Content and Form of Application Submission: It is critical that applicants follow the instructions in the Research (R) Instructions in the How to Apply - Application Guide except where instructed in this notice of funding opportunity to do otherwise. Conformance to the requirements in the Application Guide is required and strictly enforced. Applications that are out of compliance with these instructions may be delayed or not accepted for review.

Page Limitations: All page limitations described in the How to Apply- Application Guide and the Table of Page Limits must be followed.

Instructions for Application Submission: The following section supplements the instructions found in the How to Apply- Application Guide and should be used for preparing an application to this NOFO.

• SF424(R&R) Cover: All instructions in the How to Apply - Application Guide must be followed.

• SF424(R&R) Project/Performance Site Locations: All instructions in the How to Apply- Application Guide must be followed.

• SF424(R&R) Other Project Information: All instructions in the How to Apply- Application Guide must be followed.

• SF424(R&R) Senior/Key Person Profile: All instructions in the How to Apply- Application Guide must be followed.

• R&R or Modular Budget: All instructions in the How to Apply- Application Guide must be followed.

• R&R Subaward Budget: All instructions in the How to Apply-Application Guide must be followed.

• PHS 398 Cover Page Supplement: All instructions in the How to Apply- Application Guide must be followed.

• PHS 398 Research Plan: All instructions in the How to Apply- Application Guide must be followed, with the following additional instructions:

  Research Strategy: Applicants should address the following topics as they pertain to the proposed research project:

  • Importance of the Research (Significance and Innovation): Applicants should address why the chosen target or pathway is significant for the proposed chemical probe/drug discovery project, and whether the probe will provide new insight into important cancer targets and processes. Preliminary data or published information demonstrating the significance of the target or pathway should be included. Any existing modulators for the target should be mentioned, and a rationale for doing additional development of small molecules that address the target should be delineated. Innovative aspects of studying the proposed target or pathway, and of the proposed assay(s) and screening scheme should be highlighted.

  • Rigor and Feasibility (Approach): Applicants are highly encouraged to define the scope of the proposed research project via the drug discovery stages outlined in Section I and to propose a timeline and budget in keeping with the scope. Preliminary data are required to substantiate the proposed scope. For instance, if a project for HTS through hit validation is proposed (Stages 2 and 3), preliminary data should address the topics mentioned below under Assay Development, as appropriate to the project. The application should include a flow diagram to outline all critical steps in sequential and/or parallel manners with appropriate benchmarks and timelines for whichever stage(s) of discovery are proposed.

    Detailed guidelines for the three stages to be addressed in the Approach Section are as follows:

    1) Assay development: To increase the overall likelihood of a successful assay development project, applicants should include the following topics in the research plan:

    • Explanation of the scale of the proposed assay campaign (i.e., low-, moderate- or high-throughput), in keeping with the selected target. If available, include preliminary data supporting the choice of the primary screening assay for the specific target.
    • Reproducibility of the primary screening assay in a low-to-moderate throughput setting. If the goal is to perform a high-throughput screen, the research plan should include experiments to test whether the primary assay is sufficiently reproducible and robust for adaptation to an automated, high-throughput screening approach using a diverse and significant number of compounds (e.g., a collection of FDA-approved drugs or other bioactive molecules). Possible considerations when designing and characterizing a primary screening assay include effects of time, temperature and protein concentration; tolerance to the effect of DMSO at 0.1-1%; and reproducibility between plates and day-to-day experiments. Additional considerations for HTS are simplicity of the readout and ability for the primary assay to be miniaturized into formats such as 96-, 384-, or 1536-well automated screens.
    • Experiments to test predictability and reproducibility of responses to known compounds or other control conditions and a clear threshold between positive and negative responses. In general, an acceptable lower limit of Z’ factor is 0.5 for a robust single-concentration HTS that corresponds to a combination of Signal-to-Basal Ratio (S/B) of 4 with a Coefficient of Variation (CV) of 10%. Assay robustness will improve if multiple compound test concentrations or kinetic recordings are used in HTS.
    • Availability of reagents necessary to perform the screen, such as enzyme indicators, chemicals necessary for reagent readout, and the capacity to generate sufficient reaction substrates (DNA, RNA, protein, or enzyme substrates).
    • If achievable, pilot screening for target validation from a small compound library (e.g., LOPAC1280), or other bioactive molecules should be proposed as a highly desirable validation step for translation of the assay to HTS. Alternatively, dose-ranging studies for a limited set of agents having known interaction with the proposed target should be proposed. The intent of this characterization is to demonstrate the ability of the assay to pharmacologically distinguish structurally diverse compounds.

    2) Primary Screen implementation: Applications proposing to perform a screen of any scale should include preliminary data to address the topics described above in the “Assay Development” section. In addition, the topics mentioned under “Screen Implementation” in Section I should be considered when writing the Approach section. Specifically, applications are expected to include:

    • Preliminary data on the primary screening assay performance as tested in a small-scale pilot screen, including a scatter plot with a Z’ factor above 0.5 (see Assay Development section above).
    • Discussion of the following parameters: anticipated number of hits; feasibility of assessing hits (particularly large numbers); test concentrations; cut-off concentration; reagent availability; equipment used in configuring the assay; strategy to characterize initial active compounds.
    • A rationale for the size and selection of the library of compounds to be screened regardless of the scale of the screen. In certain cases, it may be advantageous to utilize focused libraries of compounds with specific consideration of assay targets, known bioactivities, privileged scaffolds, representative diversity sets, drug repurposing, and/or the ease of follow-up synthetic chemistry for SAR expansion. The compounds should be maintained under strict storage conditions and meet a set of quality control restrictions on solubility, the number of reactive groups, and compound size.

    3) Hit validation: The Approach section of the grant application is expected to include the following:

    • A clear rationale for the chosen hit validation scheme, considering the number of hits expected from the primary screen, anticipated false positives inherent in the primary assay format, and/or necessary assays to test the biology or physiology of the target.
    • Availability and suitability of secondary and counter-screen assays as appropriate for the scale of the screening campaign or plans to develop and characterize them. Preliminary data demonstrating feasibility of hit validation assays will strengthen the application.
    • Definition of the specific criteria that a compound must meet to be considered a probe or pre-therapeutic lead for the project.
    • A plan to conduct advanced cheminformatics analysis and medicinal chemistry inspection to prioritize the hit set.
    • Investigators may propose to confirm hit compounds by testing powder samples and commercially available analog compounds. If commercial analogs are available, investigators may propose preliminary SAR studies. Investigators should verify the structure of hits using a combination of analytical methods and, if possible, perform re-synthesis of select hits.
    • Critical path: The application should include a flow diagram to outline all critical steps in sequential and/or parallel manners with appropriate benchmarks and timelines for whichever stage(s) of discovery are proposed. The flow diagram may include but is not limited to: the primary assay to be implemented or previously implemented; alternative assay or approach (e.g., computational docking) to ensure success of the screening; hit selection criteria; cheminformatic analysis; chemical tractability assessment; all follow-up assays to validate screening hits; and assay responsibilities of each party involved in the project.
    • A timeline is strongly encouraged, either as a separate figure or as part of the flow diagram.

  • Resource Sharing Plan: Individuals are required to comply with the instructions for the Resource Sharing Plans as provided in the How to Apply- Application Guide.

  • Other Plan(s): All instructions in the How to Apply-Application Guide must be followed, with the following additional instructions: All applicants planning research (funded or conducted in whole or in part by NIH) that results in the generation of scientific data are required to comply with the instructions for the Data Management and Sharing Plan. All applications, regardless of the amount of direct costs requested for any one year, must address a Data Management and Sharing Plan. The following data generated or developed under this NOFO are expected to be released to PubChem: (1) primary assay data from high throughput screening (HTS), (2) data generated in all post-HTS follow-up assays, (3) protocols for assays implemented, and (4) the chemical structure of compounds tested. Applications should include a statement of willingness to deposit the aforementioned data to PubChem within the Data Management and Sharing Plan (DMS Plan) in the Other Plan(s) section of the application, consistent with achieving the goals of this program.

  • Appendix: Only limited Appendix materials are allowed. Follow all instructions for the Appendix as described in the How to Apply- Application Guide. No publications or other material, with the exception of blank questionnaires or blank surveys, may be included in the Appendix.

  • PHS Human Subjects and Clinical Trials Information: When involving human subjects research, clinical research, and/or NIH-defined clinical trials (and when applicable, clinical trials research experience) follow all instructions for the PHS Human Subjects and Clinical Trials Information form in the How to Apply- Application Guide, with the following additional instructions:

    • If you answered “Yes” to the question “Are Human Subjects Involved?” on the R&R Other Project Information form, you must include at least one human subjects study record using the Study Record: PHS Human Subjects and Clinical Trials Information form or Delayed Onset Study record.
    • Study Record: PHS Human Subjects and Clinical Trials Information: All instructions in the How to Apply- Application Guide must be followed.
    • Delayed Onset Study: Note: Delayed onset does NOT apply to a study that can be described but will not start immediately (i.e., delayed start). All instructions in the How to Apply- Application Guide must be followed.

  • PHS Assignment Request Form: All instructions in the How to Apply- Application Guide must be followed.

• Foreign Organizations: Foreign (non-U.S.) organizations must follow policies described in the NIH Grants Policy Statement, and procedures for foreign organizations described throughout the How to Apply- Application Guide.

3. Unique Entity Identifier and System for Award Management (SAM): See Part 2. Section III.1 for information regarding the requirement for obtaining a unique entity identifier and for completing and maintaining active registrations in System for Award Management (SAM), NATO Commercial and Government Entity (NCAGE) Code (if applicable), eRA Commons, and Grants.gov.

4. Submission Dates and Times: Part I. contains information about Key Dates and times. Applicants are encouraged to submit applications before the due date to ensure they have time to make any application corrections that might be necessary for successful submission. When a submission date falls on a weekend or Federal holiday, the application deadline is automatically extended to the next business day.

Organizations must submit applications to Grants.gov (the online portal to find and apply for grants across all Federal agencies). Applicants must then complete the submission process by tracking the status of the application in the eRA Commons, NIH’s electronic system for grants administration. NIH and Grants.gov systems check the application against many of the application instructions upon submission. Errors must be corrected and a changed/corrected application must be submitted to Grants.gov on or before the application due date and time. If a Changed/Corrected application is submitted after the deadline, the application will be considered late. Applications that miss the due date and time are subjected to the NIH Grants Policy Statement Section 2.3.9.2 Electronically Submitted Applications.

Applicants are responsible for viewing their application before the due date in the eRA Commons to ensure accurate and successful submission.

Information on the submission process and a definition of on-time submission are provided in the How to Apply-Application Guide.

5. Intergovernmental Review (E.O. 12372): This initiative is not subject to intergovernmental review.

6. Funding Restrictions: All NIH awards are subject to the terms and conditions, cost principles, and other considerations described in the NIH Grants Policy Statement. Pre-award costs are allowable only as described in the NIH Grants Policy Statement Section 7.9.1 Selected Items of Cost.

7. Other Submission Requirements and Information:

• Applications must be submitted electronically following the instructions described in the How to Apply Application Guide. Paper applications will not be accepted.
• Applicants must complete all required registrations before the application due date. Section III. Eligibility Information contains information about registration.
• For assistance with your electronic application or for more information on the electronic submission process, visit How to Apply – Application Guide. If you encounter a system issue beyond your control that threatens your ability to complete the submission process on-time, you must follow the Dealing with System Issues guidance. For assistance with application submission, contact the Application Submission Contacts in Section VII.

Important reminders:

• All PD(s)/PI(s) must include their eRA Commons ID in the Credential field of the Senior/Key Person Profile form.
• Failure to register in the Commons and to include a valid PD/PI Commons ID in the credential field will prevent the successful submission of an electronic application to NIH. See Section III of this NOFO for information on registration requirements.
• The applicant organization must ensure that the unique entity identifier provided on the application is the same identifier used in the organization’s profile in the eRA Commons and for the System for Award Management. Additional information may be found in the How to Apply Application Guide.
• See more tips for avoiding common errors.
• Upon receipt, applications will be evaluated for completeness and compliance with application instructions by the Center for Scientific Review and responsiveness by the NCI, NIH. Applications that are incomplete, non-compliant, and/or nonresponsive will not be reviewed.

Requests of $500,000 or more for direct costs in any year: Applicants requesting $500,000 or more in direct costs in any year (excluding consortium F&A) must contact a Scientific/Research Contact at least 6 weeks before submitting the application and follow the Policy on the Acceptance for Review of Unsolicited Applications that Request $500,000 or More in Direct Costs as described in the SF424 (R&R) Application Guide.

Mandatory Disclosure: Recipients or subrecipients must submit any information related to violations of federal criminal law involving fraud, bribery, or gratuity violations potentially affecting the federal award. See Mandatory Disclosures, 2 CFR 200.113 and NIH Grants Policy Statement Section 4.1.35. Send written disclosures to the NIH Chief Grants Management Officer listed on the Notice of Award for the IC that funded the award and to the HHS Office of Inspector Grant Self Disclosure Program at [email protected]

Post Submission Materials: Applicants are required to follow the instructions for post-submission materials, as described in the policy.

Section V. Application Review Information

1. Criteria

Only the review criteria described below will be considered in the review process. Applications submitted to the NIH in support of the NIH mission are evaluated for scientific and technical merit through the NIH peer review system.

Overall Impact: Reviewers will provide an overall impact score to reflect their assessment of the likelihood for the project to exert a sustained, powerful influence on the research field(s) involved, in consideration of the following scored review criteria and additional review criteria (as applicable for the project proposed). An application does not need to be strong in all categories to be judged likely to have a major scientific impact.

Scored Review Criteria: Reviewers will evaluate Factors 1, 2 and 3 in the determination of scientific merit, and in providing an overall impact score. In addition, Factors 1 and 2 will each receive a separate criterion score.

• Factor 1. Importance of the Research (Significance and Innovation)

  • Significance: Evaluate the importance of the proposed research in the context of current scientific challenges and opportunities, either for advancing knowledge within the field, or more broadly. Assess whether the application addresses an important gap in knowledge in the field, would solve a critical problem, or create a valuable conceptual or technical advance. Evaluate the rationale for undertaking the study, the rigor of the scientific background for the work (e.g., prior literature and/or preliminary data) and whether the scientific background justifies the proposed study.
  • Innovation: Evaluate the extent to which innovation influences the importance of undertaking the proposed research. Note that while technical or conceptual innovation can influence the importance of the proposed research, a project that is not applying novel concepts or approaches may be of critical importance for the field. Evaluate whether the proposed work applies novel concepts, methods or technologies or uses existing concepts, methods, technologies in novel ways, to enhance the overall impact of the project.
  • Specific to this NOFO: Evaluate whether the proposed work:
    • Adequately describes the novelty and importance of the biological target or pathway in cancer.
    • Delineates the rationale for developing chemical probes for the target or pathway, either as research tools to understand biology or for therapeutics development.
    • Provides a convincing rationale for identifying hits for the target or pathway, if compounds exist for the target.
    • Includes novel assays or screens for hit identification.

• Factor 2. Rigor and Feasibility (Approach)

  • Approach: Evaluate the scientific quality of the proposed work. Evaluate the likelihood that compelling, reproducible findings will result (rigor) and assess whether the proposed studies can be done well and within the timeframes proposed (feasibility).

    • Rigor: Evaluate the potential to produce unbiased, reproducible, robust data. Evaluate the rigor of experimental design and whether appropriate controls are in place. Evaluate whether the sample size is sufficient and well-justified. Assess the quality of the plans for analysis, interpretation, and reporting of results. Evaluate whether the investigators presented adequate plans to address relevant biological variables, such as sex or age, in the design, analysis, and reporting. For applications involving human subjects or vertebrate animals, also evaluate:
      • The rigor of the intervention or study manipulation (if applicable to the study design).
      • Whether outcome variables are justified.
      • Whether the results will be generalizable or, in the case of a rare disease/special group, relevant to the particular subgroup.
      • Whether the sample is appropriate and sufficiently diverse to address the proposed question(s). For applications involving human subjects, including clinical trials, assess the adequacy of inclusion plans as appropriate for the scientific goals of the research. Considerations of appropriateness may include disease/condition/behavior incidence, prevalence, or population burden, population representation, and/or current state of the science.
    • Feasibility: Evaluate whether the proposed approach is sound and achievable, including plans to address problems or new challenges that emerge in the work. For proposed studies in which feasibility may be less certain, evaluate whether the uncertainty is balanced by the potential for major advances. For applications involving human subjects, including clinical trials, evaluate the adequacy and feasibility of the plan to recruit and retain an appropriately diverse population of participants. Additionally, evaluate the likelihood of successfully achieving the proposed enrollment based on age, race, ethnicity, and sex. For clinical trial applications, evaluate whether the study timeline and milestones are feasible.

  • Specific to this NOFO: Evaluate how well the critical path diagram outlines critical steps, appropriate benchmarks and timelines for the stage(s) of discovery that are proposed. Evaluate the strength of the preliminary data in supporting the stage(s) of discovery research proposed in the aims. If animal studies are included, assess whether they are well-justified, necessary, and appropriate for early stages of drug discovery.

    Stage-specific evaluation:

    1. For development of the primary screen assay(s) and testing in an initial pilot screen, evaluate:
       • Appropriateness of the design and scale of the proposed assay campaign for the selected target.
       • Whether reproducibility and robustness of the primary assay are sufficiently addressed.
       • If a small-scale, pilot screen is proposed, assess appropriateness of the scale of the screen and the proposed library appropriate for the target.
    2. For primary screen implementation, evaluate:
       • Whether preliminary data, such as a small-scale pilot screen, support a proposed larger-scale screen.
       • The feasibility of identifying and assessing the proposed number of hits, the plan for agent availability, and the strategy to characterize the initial active compounds.
       • The size and selection of the library.
    3. For hit validation, evaluate:
       • How well justified the hit validation scheme is, considering the number of hits from the primary screen, anticipated false positives inherent in the primary assay format, and/or assays to test the biology or physiology of the target.
       • How well the criteria for a validated hit are addressed and rationalized.

• Factor 3. Expertise and Resources (Investigator(s) and Environment)

  • Investigator(s): Evaluate whether the investigator(s) have demonstrated background, training, and expertise, as appropriate for their career stage, to conduct the proposed work. For Multiple Principal Investigator (MPI) applications, assess the quality of the leadership plan to facilitate coordination and collaboration.
  • Environment: Evaluate whether the institutional resources are appropriate to ensure the successful execution of the proposed work.
  • Specific to this NOFO: Evaluate the investigators’ knowledge of the biological target, pathway, and disease area, and the adequacy of their experience in performing the proposed discovery research stages.

Additional Review Criteria: As applicable for the project proposed, reviewers will consider the following additional items while determining scientific and technical merit, but will not give criterion scores for these items, and should consider them in providing an overall impact score.

• Protections for Human Subjects: For research that involves human subjects but does not involve one of the categories of research that are exempt under 45 CFR Part 46, evaluate the justification for involvement of human subjects and the proposed protections from research risk relating to their participation according to the following five review criteria: 1) risk to subjects; 2) adequacy of protection against risks; 3) potential benefits to the subjects and others; 4) importance of the knowledge to be gained; and 5) data and safety monitoring for clinical trials. For research that involves human subjects and meets the criteria for one or more of the categories of research that are exempt under 45 CFR Part 46, evaluate: 1) the justification for the exemption; 2) human subjects involvement and characteristics; and 3) sources of materials. For additional information on review of the Human Subjects section, please refer to the Guidelines for the Review of Human Subjects.
• Vertebrate Animals: When the proposed research includes Vertebrate Animals, evaluate the involvement of live vertebrate animals according to the following criteria: (1) description of proposed procedures involving animals, including species, strains, ages, sex, and total number to be used; (2) justifications for the use of animals versus alternative models and for the appropriateness of the species proposed; (3) interventions to minimize discomfort, distress, pain and injury; and (4) justification for euthanasia method if NOT consistent with the AVMA Guidelines for the Euthanasia of Animals. For additional information on review of the Vertebrate Animals section, please refer to the Worksheet for Review of the Vertebrate Animals Section.
• Biohazards: When the proposed research includes Biohazards, evaluate whether specific materials or procedures that will be used are significantly hazardous to research personnel and/or the environment, and whether adequate protection is proposed.
• Resubmissions: As applicable, evaluate the full application as now presented.
• Renewals: As applicable, evaluate the progress made in the last funding period.
• Revisions: As applicable, evaluate the appropriateness of the proposed expansion of the scope of the project.

Additional Review Considerations: As applicable for the project proposed, reviewers will consider each of the following items, but will not give scores for these items, and should not consider them in providing an overall impact score.

• Authentication of Key Biological and/or Chemical Resources: For projects involving key biological and/or chemical resources, evaluate the brief plans proposed for identifying and ensuring the validity of those resources.
• Budget and Period of Support: Evaluate whether the budget and the requested period of support are fully justified and reasonable in relation to the proposed research.

2. Review and Selection Process

Applications will be evaluated for scientific and technical merit by (an) appropriate Scientific Review Group(s) convened by Center for Scientific Review, in accordance with NIH peer review policy and procedures, using the stated review criteria. Assignment to a Scientific Review Group will be shown in the eRA Commons.

As part of the scientific peer review, all applications will receive a written critique.

Applications may undergo a selection process in which only those applications deemed to have the highest scientific and technical merit (generally the top half of applications under review) will be discussed and assigned an overall impact score.

Applications will be assigned on the basis of established PHS referral guidelines to the appropriate NIH Institute or Center. Applications will compete for available funds with all other recommended applications submitted in response to this NOFO. Following initial peer review, recommended applications will receive a second level of review by the appropriate national Advisory Council or Board. The following will be considered in making funding decisions:

• Scientific and technical merit of the proposed project as determined by scientific peer review.
• Availability of funds.
• Relevance of the proposed project to program priorities.

If the application is under consideration for funding, NIH will request "just-in-time" information from the applicant as described in the NIH Grants Policy Statement Section 2.5.1. Just-in-Time Procedures. This request is not a Notice of Award nor should it be construed to be an indicator of possible funding.

Prior to making an award, NIH reviews an applicant’s federal award history in SAM.gov to ensure sound business practices. An applicant can review and comment on any information in the Responsibility/Qualification records available in SAM.gov. NIH will consider any comments by the applicant in the Responsibility/Qualification records in SAM.gov to ascertain the applicant’s integrity, business ethics, and performance record of managing Federal awards per 2 CFR Part 200.206 “Federal awarding agency review of risk posed by applicants.” This provision will apply to all NIH grants and cooperative agreements except fellowships.

3. Anticipated Announcement and Award Dates

After the peer review of the application is completed, the PD/PI will be able to access his or her Summary Statement (written critique) via the eRA Commons. Refer to Part 1 for dates for peer review, advisory council review, and earliest start date.

Information regarding the disposition of applications is available in the NIH Grants Policy Statement Section 2.4.4 Disposition of Applications.

Section VI. Award Administration Information

1. Award Notices

A Notice of Award (NoA) is the official authorizing document notifying the applicant that an award has been made and that funds may be requested from the designated HHS payment system or office. The NoA is signed by the Grants Management Officer and emailed to the recipient’s business official.

In accepting the award, the recipient agrees that any activities under the award are subject to all provisions currently in effect or implemented during the period of the award, due Department regulations and policies in effect at the time of the award, and applicable statutory provisions.

Recipients must comply with any funding restrictions described in Section IV.6. Funding Restrictions. Any pre-award costs incurred before receipt of the NoA are at the applicant's own risk. For more information on the Notice of Award, please refer to the NIH Grants Policy Statement Section 5. The Notice of Award and NIH Grants & Funding website, see Award Process.

Institutional Review Board or Independent Ethics Committee Approval: Recipient institutions must ensure that protocols are reviewed by their IRB or IEC. To help ensure the safety of participants enrolled in NIH-funded studies, the recipient must provide NIH copies of documents related to all major changes in the status of ongoing protocols.

2. Administrative and National Policy Requirements

The following Federal wide and HHS-specific policy requirements apply to awards funded through NIH:

• The rules listed at 2 CFR Part 200, Uniform Administrative Requirements, Cost Principles, and Audit Requirements for Federal Awards.
• All NIH grant and cooperative agreement awards include the NIH Grants Policy Statement as part of the terms and conditions in the Notice of Award (NoA). The NoA includes the requirements of this NOFO. For these terms of award, see the NIH Grants Policy Statement Part II: Terms and Conditions of NIH Grant Awards, Subpart A: General and Part II: Terms and Conditions of NIH Grant Awards, Subpart B: Terms and Conditions for Specific Types of Grants, Recipients, and Activities.
• If a recipient receives an award, the recipient must follow all applicable nondiscrimination laws. The recipient agrees to this when registering in SAM.gov. The recipient must also submit an Assurance of Compliance (HHS-690). To learn more, see the Laws and Regulations Enforced by the HHS Office for Civil Rights website. HHS recognizes that NIH research projects are often limited in scope for many reasons that are nondiscriminatory, such as the principal investigator’s scientific interest, funding limitations, recruitment requirements, and other considerations. Thus, criteria in research protocols that target or exclude certain populations are warranted where nondiscriminatory justifications establish that such criteria are appropriate with respect to the health or safety of the subjects, the scientific study design, or the purpose of the research. For additional guidance regarding how the provisions apply to NIH grant programs, please contact the Scientific/Research Contact that is identified in Section VII under Agency Contacts of this NOFO.
• All federal statutes and regulations relevant to federal financial assistance, including those highlighted in NIH Grants Policy Statement Section 4 Public Policy Requirements, Objectives and Other Appropriation Mandates.
• Recipients are responsible for ensuring that their activities comply with all applicable federal regulations. NIH may terminate awards under certain circumstances. See 2 CFR Part 200.340 Termination and NIH Grants Policy Statement Section 8.5.2 Remedies for Noncompliance or Enforcement Actions: Suspension, Termination, and Withholding of Support.

Cooperative Agreement Terms and Conditions of Award: Not Applicable

3. Data Management and Sharing

Consistent with the 2023 NIH Policy for Data Management and Sharing, when data management and sharing is applicable to the award, recipients will be required to adhere to the Data Management and Sharing requirements as outlined in the NIH Grants Policy Statement. Upon the approval of a Data Management and Sharing Plan, it is required for recipients to implement the plan as described.

4. Reporting

When multiple years are involved, recipients will be required to submit the Research Performance Progress Report (RPPR) annually and financial statements as required in the NIH Grants Policy Statement Section 8.4.1 Reporting. To learn more about post-award monitoring and reporting, see the NIH Grants & Funding website, see Post-Award Monitoring and Reporting.

A final RPPR, invention statement, and the expenditure data portion of the Federal Financial Report are required for closeout of an award, as described in the NIH Grants Policy Statement Section 8.6 Closeout. NIH NOFOs outline intended research goals and objectives. Post award, NIH will review and measure performance based on the details and outcomes that are shared within the RPPR, as described at 2 CFR Part 200.301.

Section VII. Agency Contacts

We encourage inquiries concerning this funding opportunity and welcome the opportunity to answer questions from potential applicants.

Application Submission Contacts:

• eRA Service Desk (Questions regarding ASSIST, eRA Commons, application errors and warnings, documenting system problems that threaten submission by the due date, and post-submission issues)
  • Finding Help Online: https://www.era.nih.gov/need-help (preferred method of contact)
  • Telephone: 301-402-7469 or 866-504-9552 (Toll Free)
• General Grants Information (Questions regarding application instructions, application processes, and NIH grant resources)
  • Email: [email protected] (preferred method of contact)
  • Telephone: 301-480-7075
• Grants.gov Customer Support (Questions regarding Grants.gov registration and Workspace)
  • Contact Center Telephone: 800-518-4726
  • Email: [email protected]

Scientific/Research Contact(s):

• For projects not involving cancer immunotherapy:
  • Suzanne Forry, Ph.D. National Cancer Institute (NCI) Telephone: 240-276-5922 Email: [email protected]
• For projects that involve immunotherapy:
  • Connie Sommers, Ph.D. National Cancer Institute (NCI) Telephone: 240-276-7187 Email: [email protected]

Peer Review Contact(s): Examine your eRA Commons account for review assignment and contact information (information appears two weeks after the submission due date).

Financial/Grants Management Contact(s):

• Shane Woodward National Cancer Institute (NCI) Telephone: 240-276-6303 Email: [email protected]

Section VIII. Other Information

Recently issued trans-NIH policy notices may affect your application submission. A full list of policy notices published by NIH is provided in the NIH Guide for Grants and Contracts. All awards are subject to the terms and conditions, cost principles, and other considerations described in the NIH Grants Policy Statement.

Authority and Regulations: Awards are made under the authorization of Sections 301 and 405 of the Public Health Service Act as amended (42 USC 241 and 284) and under Federal Regulations 42 CFR Part 52 and 2 CFR Part 200.

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