D 75N95021Q00018_SOW.pdf

STATEMENT OF WORK

GENERAL INFORMATION

Title of Project:

Evaluation of Drug Efficacy in a Rodent Model of Neuropathic Pain

Project Purpose or Objectives:

Replication studies are needed to evaluate/confirm findings from previous research investigations. The purpose of this order is to evaluate the efficacy of a nanoparticle drug formulation compared to free drug in a rodent neuropathic pain model.

Background Information:

There is an unmet medical need for safer, less addictive, and more effective solutions for chronic pain management. Current methods involving the prescription of opioids to patients with chronic pain has led to an unsustainable and deadly epidemic of addiction. Alternative methods of treating pain are therefore required to mitigate opioid substance abuse and addiction.

In the body, pain is transmitted through the nervous system via the activity of G-protein coupled receptors (GPCRs) found on neurons in the central and peripheral nervous systems including the brain and spinal cord. GPCRs can function at the surface of these cells, or in internalized compartments called endosomes. These endosomal GPCRs (eGPCRs) have recently been implicated in the transmission of pain, or nociception, and include the substance P neurokinin 1 receptor (NK1R) and calcitonin gene related peptide (CGRP) calcitonin- like receptor (CLR), which are co-expressed by a population of spinal neurons, and protease activated receptor 2 (PAR2), which is expressed by some nociceptors. By specifically targeting eGPCRs, it is hypothesized that superior pain relief can be achieved via particular endosomal targets which have been demonstrated to be responsible for ongoing signaling in chronic pain and which, due to their compartmentalized location, may not have been optimally targeted by previous clinical agents. Selective delivery of small molecule antagonists can be achieved with encapsulation into an acid labile nanoparticle that breaks down inside the acidic compartment of the endosome to release the antagonist, specifically targeting the eGPCR signaling of the nociceptive transmission pathway. This is a non-opioid treatment for chronic pain that could potentially bring relief to patients as a substitute for potentially harmful and habit-forming opioids.

The object of this project is to establish the efficacy of a nanoparticle formulation of aprepitant by replicating previous research investigations that compared the nanoparticle formulation to a free drug formulation in a rodent model of neuropathic pain. This requirement replicates work by Prof Nigel Bunnett (NYU) et al in this model shared with NCATS.

Period of Performance:

The period of performance is 12 months beginning on or shortly after the date of award.

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 requirements described below.

The scope of this project is for the contractor to execute the neuropathic pain model in adult rodents, treat animals with the compounds, and evaluate drug efficacy at different doses by measuring evoked and spontaneous pain.

The contractor shall perform a Mouse SNS (Sural Nerve Spared) model of neuropathic pain with use of von Frey Hairs (vFH) for assessment of mechanical allodynia. The specific methods are listed below.

Methods:

• Surgery = SNS = Sural spared, Tibial transected, Peroneal transected

• Mouse strain C57BL/6

• Drug delivery intrathecally (i.t.) by direct injection and not via catheter: test article and positive control to be administered by direct i.t. injection at 10 days post-SNS (5 µL volume used in mouse) to the L4/5 region of the spinal cord

• vFH as readout of mechanical allodynia to be measured every 30 min for at least 10 hours.

• Nanoparticles (NPs) will be freshly supplied for these studies and must be used within 4 weeks of the date of manufacture (not delivery/formulation)

• First ‘gating’ study must be to demonstrate the efficacy of a high concentration of free aprepitant (e.g. 1 µM), dosed by direct intrathecal injection, prior to evaluating nanoparticles. This is considered to be an essential positive control for utility of model for NK1 pharmacology and successful i.t. delivery in line with the extensive NK1 target literature.

• Test dose of nanoparticles: 5 µL volume of a 500 nM nanoparticle solution (can also extend to 100 / 300 /1000 nM for dose-response purposes)

• Comparator dose of free aprepitant (AP): 5 µL volume of a 500 nM AP solution (can also extend for dose-response and differentiation purposes e.g. up to 1 µM)

STUDY PROTOCOL

The contractor will draft a study that will be reviewed and approved by the Government. This protocol must be approved by the Government prior to the beginning of any research activity.

The study protocol may be later amended as needed, by agreement of the contractor and Government, to allow for changes based on early data, formulation analysis, or other events.

DELIVERABLES

The contractor shall provide a final report that includes the raw data, data analyses, a summation of the work performed, and results obtained. This report shall contain, but not be limited to, the following information: Material and Methods, Animals, Doses treated, Test materials, Experimental design, Statistical analysis, Study results, Conclusions, Tables, Figures, and a Copy of all raw data. This report shall be in sufficient detail, as determined by the Government, to comprehensively describe the achieved results.

The report shall be submitted via email to the NCATS Point(s) of Contact designated on the resultant award.

REPORTING REQUIREMENTS

Contractor will provide bi-weekly updates on the progress of the project, or more often as appropriate and/or requested by the Government. Updates shall be submitted via email to the NCATS Point(s) of Contact designated on the resultant award.

GOVERNMENT RESPONSIBILITIES

Government will have the responsibility to provide sufficient quantities of the lead compound in order for the contractor to accomplish the necessary work.