Milestones should be composed of three basic components: 1) Desired goal, 2) Timeline or decision point by which the goal must be accomplished, and 3) Evidence that the goal has been accomplished based upon quantitative criteria.
Disclaimer: The examples shown below are for illustrative purposes ONLY. NINDS is not endorsing particular development plans or models, parameters, or cut-off values for any disease, modality, or stage of development. Therefore, NINDS expects Investigators to establish milestones that are scientifically appropriate and justified for the application.
PAR-21-122: IGNITE Neurotherapeutic Agent Characterization and In vivo Efficacy Studies (R61/R33 Clinical Trial Not Allowed).
The R61 phase is for assay (efficacy and PD) planning, preparation and refinement, whereas the R33 phase is for implementation of efficacy or PD studies. Achievement of the milestone should demonstrate that the study workflow is planned and that all assays (efficacy and PD) are ready for compound or biologic testing during the R33 phase. One or more milestones must be proposed. Some examples include:
1. Efficacy assays are ready for compound or biologic testing at the time of transition to the R33 phase, according to the following criteria:
a. Positive controls show a statistically significant and dose-dependent effect relative to the negative control on the efficacy endpoint measure, where the study design included appropriate randomization and blinding procedures, with power ≥ 80%
b. The effects of positive controls on the endpoint measure can be attenuated by ≥ 70% with a test agent (e.g., target-specific antagonist/antibody/siRNA/gene vector, etc.) in a dose-dependent manner.
2. Pharmacodynamic (PD) measures are ready for compound or biologic testing at the time of transition to the R33 phase, according to the following criteria:
a. Positive controls show a statistically significant and dose-dependent effect relative to the negative control on the PD measure, where the study design included appropriate randomization and blinding procedures, with power ≥ 80%
b. The effects of positive controls on the PD measure can be attenuated by ≥ 70% with a test agent (e.g., target-specific antagonist/antibody/siRNA/gene vector, etc.) in a dose-dependent manner
c. Positive controls alter the activity of signaling molecules immediately proximal to the therapeutic target, and an antagonist, siRNA, gene vector, etc. attenuates this effect. Antagonists, siRNA, antibodies, etc. do not attenuate non-specific activation of the target
d. The PD measure shows the required dynamic range (high-low value), precision (%) and sensitivity (lowest reliable value) for the assay in question.
3. Compounds and biologics to be tested have been characterized for biophysical or physico-chemical characteristics and demonstrate properties appropriate for the reliable interpretation of efficacy and PD testing at the time of transition to the R33 phase. These characteristics include:
a. Pharmacokinetic characteristics (t1/2 ≥ X, Cmax ≥ X, AUC ≥ X, brain:plasma ≥ X, oral bioavailability ≥ X) that would allow sufficient systemic and brain exposure to provide a concentration ≥ 2x IC50 at the site of action during the time of testing, or preliminary viral vector or cell distribution
b. Biophysical/physicochemical characteristics that would allow reliable interpretation of efficacy and PD data:
i. Purity of protein produced in E.coli > 95% via SDS-PAGE or SEC-HPLC
ii. Purity of protein 88% via RP-HPLC with no more than 12% total product-related impurities
iii. Identity of protein or gene vector established by Western blot or dot blot
iv. Purity of small molecule > 95% via LC/MS
v. Solubility ≥ Xmg/mL, formulation feasible for test species at Xml/kg.
PAR-21-123: IGNITE Development and Validation of Model Systems to Facilitate Neurotherapeutic Discovery (R61/R33 Clinical Trial Not Allowed).
The R61 phase will support initial development and optimization of the animal model or ex vivo system and will include a set of clear and quantitative milestones as indicators of feasibility. After completion of this phase, the project may advance to the R33 phase for any required scaling, along with completion of internal and external validation studies. One or more milestones must be proposed. Some examples include:
1. Gene expression verified (X-fold change, resulting in X-fold change in protein) and breeding efficiency sufficient for external validation activities in the R33 phase (assuming a requirement of N animals per study/3 months)
2. Endpoints reflecting disease and/or therapeutic intervention provide sufficient reproducibility and dynamic range for use in the drug discovery process.
a. Reliability: Mean data values are within X% of each other in two or more separate runs of the assay. Alternatively, mean data values obtained in two separate assays show significant correlation (0.7-1.0).
b. Dynamic range: The floor and ceiling of the assay are separated by at X standard deviations from the mean.
PAR-21-124: IGNITE Assay Development and Neurotherapeutic Agent Identification (R61/R33 Clinical Trial Not Allowed).
The R61 phase is for assay development/feasibility and the R33 phase is for implementation of iterative screening. Achievement of the milestone should demonstrate that the phase 1 is completed and that the project is ready for the R33 phase. One or more milestones must be proposed. Some examples include:
1. Developed assays will perform with the specificity and activity required for use as a compound or biologic screening method at the time of transition to the R33 phase. Activity will be demonstrated by the following:
a. The positive control inhibits enzyme activity by > 80%
b. The negative control, does not inhibit enzyme activity (< 5%)
c. Receptor negative cells do not respond to the positive control (< 5%).
2. Developed assays will perform with the signal-to-noise, precision and dynamic range required for use as a compound or biologic screening method at the time of transition to the R33 phase. Signal-to-noise, precision and dynamic range will be demonstrated by the following:
a. The Z’ score will be ≥ 0.5, based on values from at least half a plate of positive and negative controls
b. The blinded test-retest reliability (r2 ) will be ≤ 0.75 on at least 4 positive and 4 negative compounds
c. The positive control demonstrates a dose-response relationship.
3. Developed assays will perform with the accuracy and precision required for use as a compound or biologic screening method at the time of transition to the R33 phase. The reported accuracy and precision of the assay will be within 10% of a designated reference standard.