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

Timmy Mani, Fang Wang, William Eric Knabe, Anthony L. Sinn, May Khanna, Inha Jo, George E. Sandusky, George W. Sledge, David R. Jones, Rajesh Khanna, Karen E. Pollok, Samy O. Meroueh

Research output: Contribution to journalArticle

29 Citations (Scopus)

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 5 h and peak concentration of 5 μM. Similar levels of the inhibitor were detected in tumor tissue up to 10 h. 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.

Original languageEnglish (US)
Pages (from-to)2145-2155
Number of pages11
JournalBioorganic and Medicinal Chemistry
Volume21
Issue number7
DOIs
StatePublished - Apr 1 2013
Externally publishedYes

Fingerprint

Pharmacokinetics
Breast Neoplasms
Neoplasm Metastasis
Molecules
Matrix Metalloproteinases
Tumors
Proteins
Breast
Small Molecule Libraries
Inbred NOD Mouse
Fluorescence Polarization
SCID Mice
Cell adhesion
Cell Adhesion
Cell Movement
Extracellular Matrix
Half-Life
Adipose Tissue
Assays
Neoplasms

Keywords

  • Breast cancer
  • Cancer
  • In vivo
  • Metastasis
  • Protein-protein interaction inhibitors
  • Synthesis
  • Urokinase receptor
  • Virtual screening

ASJC Scopus subject areas

  • Pharmaceutical Science
  • Drug Discovery
  • Organic Chemistry
  • Molecular Medicine
  • Molecular Biology
  • Clinical Biochemistry
  • Biochemistry

Cite this

Small-molecule inhibition of the uPAR·uPA interaction : Synthesis, biochemical, cellular, in vivo pharmacokinetics and efficacy studies in breast cancer metastasis. / Mani, Timmy; Wang, Fang; Knabe, William Eric; Sinn, Anthony L.; Khanna, May; Jo, Inha; Sandusky, George E.; Sledge, George W.; Jones, David R.; Khanna, Rajesh; Pollok, Karen E.; Meroueh, Samy O.

In: Bioorganic and Medicinal Chemistry, Vol. 21, No. 7, 01.04.2013, p. 2145-2155.

Research output: Contribution to journalArticle

Mani, Timmy ; Wang, Fang ; Knabe, William Eric ; Sinn, Anthony L. ; Khanna, May ; Jo, Inha ; Sandusky, George E. ; Sledge, George W. ; Jones, David R. ; Khanna, Rajesh ; Pollok, Karen E. ; Meroueh, Samy O. / Small-molecule inhibition of the uPAR·uPA interaction : Synthesis, biochemical, cellular, in vivo pharmacokinetics and efficacy studies in breast cancer metastasis. In: Bioorganic and Medicinal Chemistry. 2013 ; Vol. 21, No. 7. pp. 2145-2155.
@article{36a9a5d569f143b3b22a12b2b6e500bb,
title = "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 5 h and peak concentration of 5 μM. Similar levels of the inhibitor were detected in tumor tissue up to 10 h. 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.",
keywords = "Breast cancer, Cancer, In vivo, Metastasis, Protein-protein interaction inhibitors, Synthesis, Urokinase receptor, Virtual screening",
author = "Timmy Mani and Fang Wang and Knabe, {William Eric} and Sinn, {Anthony L.} and May Khanna and Inha Jo and Sandusky, {George E.} and Sledge, {George W.} and Jones, {David R.} and Rajesh Khanna and Pollok, {Karen E.} and Meroueh, {Samy O.}",
year = "2013",
month = "4",
day = "1",
doi = "10.1016/j.bmc.2012.12.047",
language = "English (US)",
volume = "21",
pages = "2145--2155",
journal = "Bioorganic and Medicinal Chemistry",
issn = "0968-0896",
publisher = "Elsevier Limited",
number = "7",

}

TY - JOUR

T1 - Small-molecule inhibition of the uPAR·uPA interaction

T2 - Synthesis, biochemical, cellular, in vivo pharmacokinetics and efficacy studies in breast cancer metastasis

AU - Mani, Timmy

AU - Wang, Fang

AU - Knabe, William Eric

AU - Sinn, Anthony L.

AU - Khanna, May

AU - Jo, Inha

AU - Sandusky, George E.

AU - Sledge, George W.

AU - Jones, David R.

AU - Khanna, Rajesh

AU - Pollok, Karen E.

AU - Meroueh, Samy O.

PY - 2013/4/1

Y1 - 2013/4/1

N2 - 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 5 h and peak concentration of 5 μM. Similar levels of the inhibitor were detected in tumor tissue up to 10 h. 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.

AB - 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 5 h and peak concentration of 5 μM. Similar levels of the inhibitor were detected in tumor tissue up to 10 h. 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.

KW - Breast cancer

KW - Cancer

KW - In vivo

KW - Metastasis

KW - Protein-protein interaction inhibitors

KW - Synthesis

KW - Urokinase receptor

KW - Virtual screening

UR - http://www.scopus.com/inward/record.url?scp=84875221885&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84875221885&partnerID=8YFLogxK

U2 - 10.1016/j.bmc.2012.12.047

DO - 10.1016/j.bmc.2012.12.047

M3 - Article

C2 - 23411397

AN - SCOPUS:84875221885

VL - 21

SP - 2145

EP - 2155

JO - Bioorganic and Medicinal Chemistry

JF - Bioorganic and Medicinal Chemistry

SN - 0968-0896

IS - 7

ER -