The National Institute of Neurologic Disorders and Stroke (NINDS) Preclinical Screening Platform for Pain (PSPP), a program within the NIH Helping to End Addiction Long-termSM, or NIH HEAL InitiativeSM, aims to accelerate the development of novel non-opioid, non-addictive therapeutics for pain. To support the PSPP goals, PsychoGenics Inc. was awarded a contract to screen and profile these novel therapeutics and to validate new endpoints and models. PSPP employs a tiered approach to evaluation of assets. In Tier 1, assets are screened in cell-based functional assays to assess activity at opioid receptors and other receptors associated with abuse liability. Also, in Tier 1, the pharmacokinetic (PK) profile of the asset in both plasma and brain is determined. In Tier 2, a side effect profile is assessed using an accelerating rotarod and modified Irwin test. Subsequently, assets are evaluated using evoked and non-evoked pain endpoints in two pain models: 1) the plantar incision model, representative of acute to sub-chronic pain mechanisms and 2) the L5/L6 spinal nerve ligation (SNL) model, representative of persistent pain mechanisms. Finally, in Tier 3, assets are evaluated in vivo for abuse liability and in disease specific pain models. This tiered approach to evaluation of assets will be illustrated using a representative example that has been screened in Tier 1 in the in vitro assays and PK, and has been profiled in Tier 2 on rotarod performance and in plantar incision and L5/L6 SNL models as well as in the intravenous self-administration model in Tier 3, enabling further evaluation in disease specific pain models within Tier 3. Together, these data demonstrate the merits of evaluating promising pain assets rigorously in a tiered approach and highlight efforts to enhance novelty and reproducibility within the NINDS PSPP program to support the goal of identifying novel non-opioid, non-addictive pain therapeutics.