Attachment_1-Statement_of_Work.pdf

STATEMENT OF WORK (SOW)

GENERAL INFORMATION

Title of Project:

Testing of compounds for off-target activity in a broad panel of safety-related targets

Project Purpose or Objectives:

This order provides for testing of 5 novel small molecule compounds in a broad panel of 174 safety-related biological targets. Such testing will inform on the activity of these compounds at critical, safety-related biological targets, and will help predict safety-related adverse events that might occur as compounds are advanced into the clinic.

Background Information:

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. The National Center for Advancing Translational Sciences (NCATS) is a translational science center that prides itself on its tremendously productive pipeline and is innovative in a number of ways. NCATS brings together a diverse range of scientists, including medicinal chemists, biologists, toxicologists, and engineers, in order to ultimately translate basic science into real products and services that help improve people’s lives. Included in this process is the development of unique small molecules as potential anticancer agents.

Prior work at NCATS has produced a series of small molecule IRAK4 / FLT3 kinase dual inhibitors, including NCGC00371481 and its structural analogs. Compounds such as ‘1481 have shown promising results in cellular and mouse models of myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML). Further optimization of the biochemical, physiochemical and pharmacokinetic properties of these compounds is required, and may eventually produce a clinical candidate that could be used to treat cancers such as MDS and / or AML.

On a separate but related program, prior work at NCATS has led to the discovery of novel small molecule inhibitors of the kinase enzymes LATS1 and LATS2. Inhibitors of these targets may be useful in wound healing, tissue regeneration, and other related areas.

In order to fully assess and optimize the biological properties of compounds on these two programs, NCATS needs to understand not only their ‘on-target activity’ (that is, their inhibition of the desired targets mentioned above), but also their ‘off-target activity’, meaning any undesired activity at other biological targets. In particular, NCATS needs to understand if they have significant activity at off-targets that are important for safety. By monitoring potential activity at these targets in vitro, NCATS can predict the occurrence of safety-related adverse events that might occur once compounds are advanced into clinical testing. This data will thus

RFQ Number 75N95019Q00111 Attachment 1 help NCATS to identify and ultimately optimize out any unwanted activity, thus allowing NCATS to prioritize the safest compounds for advancement into the clinic.

Testing of five (5) small molecule compounds against a broad panel of 174 safety-related targets will allow us to prioritize safer compounds for ultimate advancement into human clinical trials.

Period of Performance:

Period of performance is within three (3) months from the contract award and receipt of first batch of material.

SCOPE OF WORK

General 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 Statement of Work below:

Contractor shall provide the following services:

1. Perform experiments using established binding assays testing each of five (5) small molecule compounds for activity against at least 174 safety-related targets, including all of the following GPCR, enzyme and ion channel targets:

A1 Human Adenosine GPCR Binding (Antagonist Radioligand) Assay A2A Human Adenosine GPCR Binding (Agonist Radioligand) Assay A2B Human Adenosine GPCR Binding (Antagonist Radioligand) Assay A3 Human Adenosine GPCR Binding (Agonist Radioligand) Assay Adenosine Guinea Pig Transporter Binding (Antagonist Radioligand) Assay alpha1A Rat Adrenoceptor GPCR Binding (Antagonist Radioligand) Assay alpha1B Rat Adrenoceptor GPCR Binding (Antagonist Radioligand) Assay alpha1D Human Adrenoceptor GPCR Binding (Antagonist Radioligand) Assay alpha2A Human Adrenoceptor GPCR Binding (Antagonist Radioligand) Assay alpha2B Human Adrenoceptor GPCR Binding (Antagonist Radioligand) Assay alpha2C Human Adrenoceptor GPCR Binding (Antagonist Radioligand) Assay beta1 Human Adrenoceptor GPCR Binding (Antagonist Radioligand) Assay beta2 Human Adrenoceptor GPCR Binding (Antagonist Radioligand) Assay beta3 Human Adrenoceptor GPCR Binding (Antagonist Radioligand) Assay NET Human Norepinephrine Transporter Binding (Antagonist Radioligand) Assay MCR Human Aldosterone NHR Binding (Agonist Radioligand) Assay AR Human Androgen NHR Binding (Agonist Radioligand) Assay AT1 Human Angiotensin GPCR Binding (Agonist Radioligand) Assay AT2 Human Angiotensin GPCR Binding (Agonist Radioligand) Assay APJ Human Apelin GPCR Binding (Agonist Radioligand) Assay

Atrial Natriuretic Factor (ANF, Non-Selective) Guinea Pig Binding Assay BB1 Human Bombesin GPCR Binding (Agonist Radioligand) Assay BB2 Human Bombesin GPCR Binding (Agonist Radioligand) Assay BB3 Human Bombesin GPCR Binding (Agonist Radioligand) Assay B1 Human Bradykinin GPCR Binding (Agonist Radioligand) Assay B2 Human Bradykinin GPCR Binding (Agonist Radioligand) Assay CGRP (CLR/RAMP1) Human Calcitonin GPCR Binding (Agonist Radioligand) Assay Cav1.2 (L-type) Rat Calcium Ion Channel Binding (Diltiazem Site) Assay Cav1.2 (L-type) Rat Calcium Ion Channel Binding (Dihydropyridine Site) Assay Cav1.2 (L-type) Rat Calcium Ion Channel Binding (Verapamil Site) Assay Cav2.2 (N-type) Rat Calcium Ion Channel Binding (Antagonist Radioligand) Assay CB1 Human Cannabinoid GPCR Binding (Antagonist Radioligand) Assay CB2 Human Cannabinoid GPCR Binding (Agonist Radioligand) Assay CCR1 Human Chemokine GPCR Binding (Agonist Radioligand) Assay CCR2 Human Chemokine GPCR Binding (Agonist Radioligand) Assay CCR5 Rhesus Macaque Chemokine GPCR Binding (Agonist Radioligand) Assay CCK1 (CCKA) Human Cholecystokinin GPCR Binding (Antagonist Radioligand) Assay CCK2 (CCKB) Human Cholecystokinin GPCR Binding (Agonist Radioligand) Assay CHT1 Rat Choline Transporter Binding (Antagonist Radioligand) Assay Rat Tubulin [3H] Colchicine Binding Assay CRF1 Human Corticotropin-Releasing Factor GPCR Binding (Agonist Radioligand) Assay D1 Human Dopamine GPCR Binding (Antagonist Radioligand) Assay D2S Human Dopamine GPCR Binding (Antagonist Radioligand) Assay D3 Human Dopamine GPCR Binding (Antagonist Radioligand) Assay D4.2 Human Dopamine GPCR Binding (Antagonist Radioligand) Assay D5 Human Dopamine GPCR Binding (Antagonist Radioligand) Assay DAT Human Dopamine Transporter Binding (Antagonist Radioligand) Assay ETA Human Endothelin GPCR Binding (Agonist Radioligand) Assay ETB Human Endothelin GPCR Binding (Agonist Radioligand) Assay EGFR Human RTK Kinase Binding (Agonist Radioligand) Assay ERalpha Human Estrogen NHR Binding (Agonist Radioligand) Assay ERbeta Human Estrogen NHR Binding (Agonist Radioligand) Assay GABA (Non-Selective) Rat Transporter Binding (Antagonist Radioligand) Assay Non-Selective Rat GABAA Ion Channel [3H] Muscimol Binding (Agonist Radioligand) Assay Non-Selective Rat GABAA Ion Channel [3H] Flunitrazepam Binding (Agonist Radioligand) Assay Non-Selective Rat GABAA Ion Channel [3H] TBOB Binding (Antagonist Radioligand) Assay GABAB (B1a/B2) Human GABAB GPCR Binding (Antagonist Radioligand) Assay GABAB (B1b/B2) Human GABAB GPCR Binding (Antagonist Radioligand) Assay Rat Cortex [3H] Gabapentin Binding Assay GHB Rat Binding (Antagonist Radioligand) Assay Glucagon Human Glucagon GPCR Binding (Agonist Radioligand) Assay GR Human Glucocorticoid NHR Binding (Agonist Radioligand) Assay Glutamate (AMPA, Non-Selective) Rat Ion Channel [3H] AMPA Binding Assay

Glutamate, Kainate Glutamate (NMDA, Non-Selective) Rat Ion Channel [3H] CGP-39653 Binding Assay Glutamate, NMDA, Glycine [3H] MDL 105,519 Binding Assay Glutamate (Non-Selective) Rat Ion Channel [3H] TCP Binding Assay Glutamate (NMDA, Non-Selective) Rat Ion Channel [3H] Ifenprodil Binding Assay mGlu2 Human Glutamate (Metabotropic) GPCR Binding (Antagonist Radioligand) Assay mGlu5 Human Glutamate (Metabotropic) GPCR Binding (Agonist Radioligand) Assay Non-Selective Rat Glycine Ion Channel [3H] Strychnine Binding Assay GLYT1 Rat Glycine Transporter Binding (Antagonist Radioligand) Assay GnRH Human Gonadotrophin-Releasing Hormone GPCR Binding (Agonist Radioligand) Assay GHSR Human Ghrelin GPCR Binding (Agonist Radioligand) Assay H1 Human Histamine GPCR Binding (Antagonist Radioligand) Assay H2 Human Histamine GPCR Binding (Antagonist Radioligand) Assay H3 Human Histamine GPCR Binding (Agonist Radioligand) Assay Imidazoline I2, Central Rat Binding (Antagonist Radioligand) Assay IP3R1 Rat IP3 Ion Channel Binding (Agonist Radioligand) Assay IR Rat RTK Kinase Binding (Agonist Radioligand) Assay Interleukin IL-1 R1 IL-6 Human Binding (Agonist Radioligand) Assay CXCR2 (Non-Selective) Human Chemokine GPCR Binding (Agonist Radioligand) Assay CXCR2 (IL-8RB) Human Chemokine GPCR Binding (Agonist Radioligand) Assay CXCR3 Human Chemokine GPCR Binding (Agonist Radioligand) Assay CXCR4 Human Chemokine GPCR Binding (Agonist Radioligand) Assay CX3CR1 Human Chemokine GPCR Binding (Agonist Radioligand) Assay CysLT1 Human Leukotriene GPCR Binding (Agonist Radioligand) Assay BLT1 (LTB4) Human Leukotriene GPCR Binding (Agonist Radioligand) Assay MCH1 Human Melanin-Conc. Hormone GPCR Binding (Agonist Radioligand) Assay MC1 Human Melanocortin GPCR Binding (Agonist Radioligand) Assay MC3 Human Melanocortin GPCR Binding (Agonist Radioligand) Assay MC4 Human Melanocortin GPCR Binding (Agonist Radioligand) Assay MC5 Human Melanocortin GPCR Binding (Agonist Radioligand) Assay MT1 Human Melatonin GPCR Binding (Agonist Radioligand) Assay MT2 Human Melatonin GPCR Binding (Agonist Radioligand) Assay VMAT (Non-Selective) Human Vesicular Monoamine Transporter Binding Assay Motilin Human Motilin GPCR Binding (Agonist Radioligand) Assay M1 Human Acetylcholine (Muscarinic) GPCR Binding (Antagonist Radioligand) Assay M2 Human Acetylcholine (Muscarinic) GPCR Binding (Antagonist Radioligand) Assay M3 Human Acetylcholine (Muscarinic) GPCR Binding (Antagonist Radioligand) Assay M4 Human Acetylcholine (Muscarinic) GPCR Binding (Antagonist Radioligand) Assay M5 Human Acetylcholine (Muscarinic) GPCR Binding (Antagonist Radioligand) Assay Non-Selective Rat Acetylcholine (Muscarinic) GPCR Binding (Agonist Radioligand) Assay NK1 Human Tachykinin GPCR Binding (Agonist Radioligand) Assay NK2 Human Tachykinin GPCR Binding (Antagonist Radioligand) Assay

NK3 Human Tachykinin GPCR Binding (Agonist Radioligand) Assay Y1 Human Neuropeptide Y GPCR Binding (Agonist Radioligand) Assay Y2 Human Neuropeptide Y GPCR Binding (Agonist Radioligand) Assay NT1 Human Neurotensin GPCR Binding (Agonist Radioligand) Assay nAChR Human Acetylcholine (Nicotinic) Ion Channel Binding Assay nAChR (alpha7) Rat Ion Channel Binding (Antagonist Radioligand) Assay nAChR (alpha7) Human Ion Channel [3H] Methyllycaconitine Binding Assay nAChR (alpha1) Human Ion Channel Binding (Antagonist Radioligand) Assay nAChR (alpha4/beta2) Rat Ion Channel Binding (Agonist Radioligand) Assay delta (DOP) Human Opioid GPCR Binding (Antagonist Radioligand) Assay kappa (KOP) Human Opioid GPCR Binding (Antagonist Radioligand) Assay mu (MOP) Human Opioid GPCR Binding (Antagonist Radioligand) Assay NOP (ORL1) Human Opioid GPCR Binding (Agonist Radioligand) Assay OT Human Vasopressin / Oxytocin GPCR Binding (Agonist Radioligand) Assay Phorbol Ester Binding Assay PAF Human Platelet-Activating Factor GPCR Binding (Agonist Radioligand) Assay PDGFR (Non-Selective) Mouse RTK Kinase Binding (Agonist Radioligand) Assay KV (Non-Selective) Rat Potassium Ion Channel [125I] alpha-Dendrotoxin Binding Assay KATP Hamster Potassium Ion Channel Binding (Antagonist Radioligand) Assay SKCa (Non-Selective) Rat Potassium Ion Channel [125I] Apamin Binding Assay hERG Human Potassium Ion Channel [3H] Astemizole Binding (Antagonist Radioligand) Assay DP2 Human Prostanoid GPCR Binding (Agonist Radioligand) Assay DP1 Human Prostanoid GPCR Binding (Agonist Radioligand) Assay EP1 Human Prostanoid GPCR Binding (Agonist Radioligand) Assay EP2 Human Prostanoid GPCR Binding (Agonist Radioligand) Assay EP4 Human Prostanoid GPCR Binding (Agonist Radioligand) Assay FP Human Prostanoid GPCR Binding (Agonist Radioligand) Assay Non-Selective Rat P2Y GPCR Binding (Agonist Radioligand) Assay RXRalpha Human Retinoid X NHR Binding (Agonist Radioligand) Assay Non-Selective Rat PDE [3H] Rolipram Binding (Antagonist Radioligand) Assay RY3 Rat Ryanodine Ion Channel Binding (Agonist Radioligand) Assay 5-HT1A Human Serotonin GPCR Binding (Agonist Radioligand) Assay 5-HT1B Rat Serotonin GPCR Binding (Antagonist Radioligand) Assay 5-HT2B Human Serotonin GPCR [3H]LSD Binding (Agonist Radioligand) Assay 5-HT2C Human Serotonin GPCR Binding (Antagonist Radioligand) Assay 5-HT3 Human Serotonin Ion Channel Binding (Antagonist Radioligand) Assay 5-HT4 Guinea Pig Serotonin GPCR Binding (Antagonist Radioligand) Assay 5-HT5A Human Serotonin GPCR Binding (Agonist Radioligand) Assay 5-HT6 Human Serotonin GPCR Binding (Agonist Radioligand) Assay 5-HT7 Human Serotonin GPCR Binding (Agonist Radioligand) Assay SET Human Serotonin Transporter Binding (Antagonist Radioligand) Assay sigma1 Human Binding (Antagonist Radioligand) Assay sigma2 Rat [3H] Ifenprodil Binding (Agonist Radioligand) Assay

Non-Selective Rat Sodium Ion Channel [3H] Batrachotoxinin Binding (Site 2) Assay sst1 Human Somatostatin GPCR Binding (Agonist Radioligand) Assay sst3 Human Somatostatin GPCR Binding (Agonist Radioligand) Assay sst2 Human Somatostatin GPCR Binding (Agonist Radioligand) Assay sst4 Human Somatostatin GPCR Binding (Agonist Radioligand) Assay sst5 Human Somatostatin GPCR Binding (Agonist Radioligand) Assay TR (Non-Selective) Rat Thyroid Hormone NHR Binding (Agonist Radioligand) Assay TRH Rat Thyrotropin-RH GPCR Binding (Agonist Radioligand) Assay TGFbeta (Non-Selective) Mouse TKL Kinase Binding (Agonist Radioligand) Assay UT (GPR14) Human Urotensin GPCR Binding (Agonist Radioligand) Assay VIP1 (VPAC1) Human VIP and PACAP GPCR Binding (Agonist Radioligand) Assay VIP2 (VPAC2) Human VIP and PACAP GPCR Binding (Agonist Radioligand) Assay V1A Human Vasopressin / Oxytocin GPCR Binding (Antagonist Radioligand) Assay V1B Human Vasopressin / Oxytocin GPCR Binding (Agonist Radioligand) Assay V2 Human Vasopressin / Oxytocin GPCR Binding (Agonist Radioligand) Assay VDR Human NHR Binding (Agonist Radioligand) Assay OX2 Human Orexin GPCR Binding (Agonist Radioligand) Assay CysLT2 Human Leukotriene GPCR Binding (Agonist Radioligand) Assay IP Human Prostanoid GPCR Binding (Agonist Radioligand) Assay PR Human Progesterone NHR Binding (Agonist Radioligand) Assay EP3 Human Prostanoid GPCR Binding (Agonist Radioligand) Assay CT Human Calcitonin GPCR Binding (Agonist Radioligand) Assay OX1 Human Orexin GPCR Binding (Agonist Radioligand) Assay Histamine H4 Non-Selective Rat P2X Ion Channel Binding (Agonist Radioligand) Assay

2. Provide a comprehensive report with the results of the single-dose duplicate testing of each of the five (5) compounds against the 174 targets.

Specific Requirements:

1. Contractor will have significant experience (more than five years) in the testing of small molecule compounds for activity against the panel of 174 targets.

2. Contractor will have access to all critical instrumentation necessary for this statement of work.

3. Contractor will be capable of testing the aforementioned five (5) compounds in all the assays described above within three (3) months.

Level of Effort:

Estimated personnel: 1 PhD level-Manager. 1 Ph.D. level scientist. 4-5 months total.

GOVERNMENT RESPONSIBILITIES

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

Contractor shall not use federal facilities. Government shall have the responsibility to review and approve the final report. Government shall have the responsibility to provide sufficient quantities of the lead compound in order for contractor to accomplish the necessary work.

REPORTING REQUIREMENTS

Contractor shall provide monthly updates on the progress of the project, or more often as appropriate. Contractor shall provide the data analysis and report for the contents of this statement of work.

OTHER CONSIDERATIONS

Data Rights:

Any data generated from this work shall be the property of NCATS.

Publications and Publicity:

Contractor shall not publish the results of this work.

Confidentiality of Information:

Contractor shall keep the results of this work confidential.