Abstract
The 6-AH family [d-Nle-X-Ile-NH-(CH2)5-CONH2; where X = various amino acids] of angiotensin IV (Ang IV) analogs binds directly to hepatocyte growth factor (HGF) and inhibit HGF's ability to form functional dimers. The metabolically stabilized 6-AH family member, d-Nle-Tyr-Ile-NH-(CH2)5-CONH2, had a t1/2 in blood of 80 min compared with the parent compound norleual [Nle-Tyr-Leu-Ψ-(CH2-NH2)3-4-His-Pro-Phe], which had a t1/2 in blood of <5 min. 6-AH family members were found to act as mimics of the dimerization domain of HGF (hinge region) and inhibited the interaction of an HGF molecule with a 3H-hinge region peptide resulting in an attenuated capacity of HGF to activate its receptor Met. This interference translated into inhibition of HGF-dependent signaling, proliferation, and scattering in multiple cell types at concentrations down into the low picomolar range. We also noted a significant correlation between the ability of the 6-AH family members to block HGF dimerization and inhibition of the cellular activity. Furthermore, a member of the 6-AH family with cysteine at position 2, was a particularly effective antagonist of HGF-dependent cellular activities. This compound suppressed pulmonary colonization by B16-F10 murine melanoma cells, which are characterized by an overactive HGF/Met system. Together, these data indicate that the 6-AH family of Ang IV analogs exerts its biological activity by modifying the activity of the HGF/Met system and offers the potential as therapeutic agents in disorders that are dependent on or possess an overactivation of the HGF/Met system.
Footnotes
This work was supported by a grant from the Adler Foundation (to J.W.H.).
J.W.W. and J.W.H. are founders and shareholders in M3 Biotechnology, LLC, which is developing pharmaceuticals based on this technology.
Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
-
ABBREVIATIONS:
- HGF
- hepatocyte growth factor
- ANG
- angiotensin
- Fmoc
- 9-fluorenylmethoxycarbonyl
- HPLC
- high-performance liquid chromatography
- HEK293
- human embryonic kidney 293
- LC-MS
- liquid chromatography-mass spectrometry
- MDCK
- Madin-Darby canine kidney
- DMEM
- Dulbecco's modified Eagle's medium
- FBS
- fetal bovine serum
- Bis-Tris
- 2-[bis(2 hydroxyethyl)amino]-2-(hydroxymethyl)propane-1,3-diol
- ψ-(CH2-NH2)
- reduced peptide bond
- Vd
- volume of distribution
- PBS
- phosphate-buffered saline
- PAGE
- polyacrylamide gel electrophoresis
- BS3
- bissulfosuccinimidyl suberate
- Nle1-Ang IV
- Nle-Tyr-Ile-His-Pro-Phe
- MTT
- 1-(4,5-dimethylthiazol-2-yl)3,5-diphenylformazan reagent
- ANOVA
- analysis of variance.
- Received September 16, 2011.
- Accepted November 29, 2011.
- Copyright © 2012 by The American Society for Pharmacology and Experimental Therapeutics
JPET articles become freely available 12 months after publication, and remain freely available for 5 years.Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page.
|