Small-molecule inhibition of the uPAR·uPA interaction: synthesis, biochemical, cellular, in vivo pharmacokinetics and efficacy studies in breast cancer metastasis.

Abstract

The uPAR·uPA protein-protein interaction (PPI) is involved in signaling and proteolytic events that promote tumor invasion and metastasis. A previous study had identified 4 (IPR-803) from computational screening of a commercial chemical library and shown that the compound inhibited uPAR·uPA PPI in competition biochemical assays and invasion cellular studies. Here, we synthesize 4 to evaluate in vivo pharmacokinetic (PK) and efficacy studies in a murine breast cancer metastasis model. First, we show, using fluorescence polarization and saturation transfer difference (STD) NMR, that 4 binds directly to uPAR with sub-micromolar affinity of 0.2 μM. We show that 4 blocks invasion of breast MDA-MB-231, and inhibits matrix metalloproteinase (MMP) breakdown of the extracellular matrix (ECM). Derivatives of 4 also inhibited MMP activity and blocked invasion in a concentration-dependent manner. Compound 4 also impaired MDA-MB-231 cell adhesion and migration. Extensive in vivo PK studies in NOD-SCID mice revealed a half-life of nearly 5h and peak concentration of 5 μM. Similar levels of the inhibitor were detected in tumor tissue up to 10h. Female NSG mice inoculated with highly malignant TMD-MDA-MB-231 in their mammary fat pads showed that 4 impaired metastasis to the lungs with only four of the treated mice showing severe or marked metastasis compared to ten for the untreated mice. Compound 4 is a promising template for the development of compounds with enhanced PK parameters and greater efficacy.

• Authors: Timmy Mani, Fang Wang, William Eric Knabe, Anthony L Sinn, May Khanna, Inha Jo, George E Sandusky, George W Sledge Jr, David R Jones, Rajesh Khanna, Karen E Pollok, Samy O Meroueh
• Journal: Bioorganic & medicinal chemistry (Bioorg Med Chem) Vol. 21 Issue 7 Pg. 2145-55 (Apr 01 2013) ISSN: 1464-3391 [Electronic] England
• PMID: 23411397 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2013 Elsevier Ltd. All rights reserved.

Chemical References

• Antineoplastic Agents
• Receptors, Urokinase Plasminogen Activator
• Small Molecule Libraries
• Urokinase-Type Plasminogen Activator

Topics

• Animals
• Antineoplastic Agents (chemistry, pharmacokinetics, pharmacology, therapeutic use)
• Breast (drug effects, metabolism, pathology)
• Breast Neoplasms (drug therapy, metabolism, pathology)
• Cell Line, Tumor
• Cell Movement (drug effects)
• Female
• Humans
• Mice
• Mice, Inbred NOD
• Mice, SCID
• Neoplasm Invasiveness (pathology)
• Neoplasm Metastasis (drug therapy, pathology)
• Protein Interaction Maps (drug effects)
• Receptors, Urokinase Plasminogen Activator (antagonists & inhibitors, metabolism)
• Small Molecule Libraries (chemistry, pharmacokinetics, pharmacology, therapeutic use)
• Urokinase-Type Plasminogen Activator (antagonists & inhibitors, metabolism)