Isolated nucleic acid molecule encoding a peptide or peptide analog comprising the SEQ ID No. 3
Inventors
Remaley, Alan T. • Demosky, Stephen J. • Stonik, John A. • Amar, Marcele J. A. • Neufeld, Edward B. • Brewer, Bryan H. • Thomas, Fairwell
Assignees
US Department of Health and Human Services
Publication Number
US-8071746-B2
Publication Date
2011-12-06
Expiration Date
2025-10-14
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Abstract
Disclosed herein are peptides or peptide analogs with multiple amphipathic α-helical domains that promote lipid efflux from cells via an ABCA1-dependent pathway. Also provided herein are methods of using multi-domain amphipathic α-helical peptides or peptide analogs to treat or inhibit dyslipidemic disorders. Methods for identifying non-cytotoxic peptides that promote ABCA1-dependent lipid efflux from cells are also disclosed herein.
Core Innovation
The invention relates to isolated peptides or peptide analogs comprising multiple amphipathic α-helical domains that promote lipid efflux from cells via an ABCA1-dependent pathway. These peptides include a first amphipathic α-helical domain exhibiting higher lipid affinity relative to a second amphipathic α-helical domain within the same peptide. The invention also encompasses methods of using these multi-domain amphipathic α-helical peptides or analogs to treat or inhibit dyslipidemic and vascular disorders, such as hyperlipidemia, coronary artery disease, and atherosclerosis.
The background addresses the problem caused by cytotoxicity associated with existing synthetic peptide mimics of apolipoproteins that promote lipid efflux from cells by non-specific, ABCA1-independent pathways. These non-specific pathways can damage cells by microsolubilization of the plasma membrane and reduce the therapeutic benefits of peptides by removing cholesterol from non-ABCA1-expressing cells. Therefore, there is a need for non-cytotoxic synthetic peptide mimics that promote specific lipid efflux through the ABCA1-dependent mechanism to provide effective treatment for cardiovascular diseases without causing cellular toxicity.
The invention solves this problem by providing peptides with multiple amphipathic α-helical domains containing at least one helix with relatively high lipid affinity and another helix with relatively low lipid affinity. This composition specifically promotes lipid efflux through the ABCA1 transporter, thereby reducing cytotoxic effects. The peptides are characterized by hydrophobic moment scores on the Eisenberg scale to quantify lipid affinity, and the difference between the high and low lipid affinity helices is at least 0.01 per residue. The invention also includes pharmaceutical compositions, methods for treating dyslipidemic and vascular disorders, and methods for identifying substantially non-cytotoxic peptides with ABCA1-dependent lipid efflux activity.
Claims Coverage
The patent discloses one independent claim focusing on an isolated nucleic acid molecule encoding a peptide or peptide analog comprising a specified amino acid sequence. There are four dependent claims elaborating features related to lipid efflux activity, peptide analogs with additional domains, and medical use.
Isolated nucleic acid molecule encoding a specific peptide sequence
An isolated nucleic acid encoding a peptide or peptide analog comprising the amino acid sequence set forth in SEQ ID NO: 3.
Peptide promoting ABCA1-dependent lipid efflux and low cytotoxicity
The isolated nucleic acid molecule encodes the peptide set forth in SEQ ID NO: 3 that promotes ATP-binding cassette transporter A1 (ABCA1)-dependent lipid efflux from cells and is substantially non-cytotoxic.
Encoding peptides with additional functional domains
The nucleic acid molecule encodes a peptide or peptide analog as claimed and includes at least one additional peptide domain.
Additional peptide domains with specified sequences
The additional peptide domain comprises sequences such as a heparin binding site, an integrin binding site, a P-selectin site, TAT HIV sequence, panning sequence, penatratin sequence, serum amyloid A (SAA) C-terminus or N-terminus sequences, LDL receptor sequences, modified 18A sequence, apoA-I Milano sequence, 6×-His sequence, or lactoferrin sequence, or combinations thereof.
The claimed invention covers a nucleic acid molecule encoding a peptide with a defined amino acid sequence that promotes ABCA1-dependent lipid efflux non-cytotoxically, with optional additional peptide functional domains, and its use in treating dyslipidemic and vascular disorders via implants.
Stated Advantages
The peptides promote specific lipid efflux from cells via an ABCA1-dependent pathway, reducing cytotoxic effects.
The peptides provide therapeutic benefit in treating and preventing dyslipidemic and vascular disorders with increased specificity and reduced cell membrane damage.
Multi-domain peptides with asymmetry in lipid affinity enhance ABCA1 transporter specificity and reduce cytotoxicity compared to peptides with uniformly high lipid affinity.
Documented Applications
Treatment and prevention of dyslipidemic and vascular disorders including hyperlipidemia, hyperlipoproteinemia, hypercholesterolemia, hypertriglyceridemia, HDL deficiency, apoA-I deficiency, coronary artery disease, atherosclerosis, thrombotic stroke, peripheral vascular disease, restenosis, acute coronary syndrome, and reperfusion myocardial injury.
Use of peptides or peptide analogs in pharmaceutical compositions for administration to subjects.
Use of nucleic acid molecules encoding peptides for gene therapy or expression in vivo to treat dyslipidemic or vascular disorders.
Methodologies for identifying non-cytotoxic peptides that promote ABCA1-dependent lipid efflux involving cytotoxicity and lipid efflux assays in vitro.
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