WO2008094905A2 - Encapsulated hdl mimetic peptides - Google Patents

Encapsulated hdl mimetic peptides Download PDF

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Publication number
WO2008094905A2
WO2008094905A2 PCT/US2008/052308 US2008052308W WO2008094905A2 WO 2008094905 A2 WO2008094905 A2 WO 2008094905A2 US 2008052308 W US2008052308 W US 2008052308W WO 2008094905 A2 WO2008094905 A2 WO 2008094905A2
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WO
WIPO (PCT)
Prior art keywords
leu
lys
giu
ala
ser
Prior art date
Application number
PCT/US2008/052308
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French (fr)
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WO2008094905A3 (en
Inventor
H. Bryan Brewer Jr.
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Lipid Sciences, Inc.
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Publication date
Priority claimed from US11/764,619 external-priority patent/US20080199398A1/en
Priority claimed from US11/861,758 external-priority patent/US20080138284A1/en
Application filed by Lipid Sciences, Inc. filed Critical Lipid Sciences, Inc.
Priority claimed from US12/021,629 external-priority patent/US20080227686A1/en
Priority claimed from US12/021,486 external-priority patent/US20080206142A1/en
Publication of WO2008094905A2 publication Critical patent/WO2008094905A2/en
Publication of WO2008094905A3 publication Critical patent/WO2008094905A3/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/10—Peptides having 12 to 20 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Liposomes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Liposomes
    • A61K9/1275—Lipoproteins; Chylomicrons; Artificial HDL, LDL, VLDL, protein-free species thereof; Precursors thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L31/16—Biologically active materials, e.g. therapeutic substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
    • A61L2300/252—Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
    • A61L2300/62—Encapsulated active agents, e.g. emulsified droplets

Definitions

  • the present invention relates to novel compositions and methods of administering these compositions to treat dyslipidemic and vascular disorders in humans and animals.
  • the compositions are vesicles encapsulating high density lipoprotein (HDL) mimetic peptides that stimulate lipid efflux and/or increase HDL levels in an animal or a human. Increased HDL levels provide numerous health benefits, particularly cardiovascular health benefits.
  • HDL high density lipoprotein
  • Cardiovascular disease is a major cause of mortality and morbidity, especially in countries of the Western world.
  • Dyslipidemic and vascular disorders affect numerous individuals throughout the world. Diseases such as hyperlipidemia, hyperlipoproteinemia, hypercholesterolemia, hypertriglyceridemia, HDL deficiency, apoAl deficiency, coronary artery disease, atherosclerosis, myocardial infarction, stroke and inflammation secondary to stroke, ischemia, ischemic stroke, thrombotic stroke, peripheral vascular disease, restenosis, thrombosis, acute coronary syndrome, and reperfusion myocardial injury cause suffering and death.
  • plasma levels of apoAl and HDL are unacceptably low, while other molecules such as low density lipoprotein (LDL) remain high. Many of these individuals are at risk for significant cardiovascular disease and heart attacks.
  • LDL low density lipoprotein
  • compositions that increase lipid efflux and HDL levels in a recipient of the composition. What is also needed are new compositions that are effective in treating these conditions or in slowing the progression of disease.
  • the present invention provides novel compositions and methods of administering these compositions to animals or humans in order to increase lipid efflux and increase HDL levels in the animal or human.
  • these compositions may be delivered orally or may be coated on stents for delivery to blood vessels.
  • the novel compositions comprise a vesicle encapsulating an HDL mimetic peptide.
  • these novel compositions are combined with a pharmaceutically acceptable carrier to make a pharmaceutical composition which may be administered to an individual to promote lipid efflux.
  • the compositions of the invention can be used to manufacture medicaments for increasing lipid efflux, increasing HDL levels, and/or treating a dyslipidemic or vascular disorder in an individual receiving the medicament.
  • compositions of the invention can be manufactured for oral administration, for coating a vascular stent, for intravenous or intramuscular administration, or for administration by any other means known in the art and/or set forth herein.
  • Administration of the compositions of the invention to individuals in need thereof can be through any appropriate means, including but not limited to oral administration or as a coating on a stent.
  • An individual who may be treated by administering the compositions of the present invention includes any individual who may benefit from an increase in lipid efflux or an increase in plasma HDL levels. Such individuals may have a dyslipidemic or vascular disease. Other individuals who may be treated have no clinical manifestation of disease but may be prone to such disease through genetic and/or environmental factors. Treatment of these individuals helps to delay or prevent the onset of dyslipidemic or vascular disease.
  • Administration of these pharmaceutical compositions may be through any means described herein and includes, but is not limited to, oral administration or as a coating on a stent.
  • Dyslipidemic and vascular disorders amenable to treatment with the compositions disclosed herein include, but are not limited to, hyperlipidemia, hyperlipoproteinemia, hypercholesterolemia, hypertriglyceridemia, HDL deficiency, apoAl deficiency, coronary artery disease, atherosclerosis, myocardial infarction, stroke and inflammation secondary to stroke, ischemia, ischemic stroke, thrombotic stroke, peripheral vascular disease, restenosis, thrombosis, acute coronary syndrome, and reperfusion myocardial injury. Accordingly, it is an object of the present invention to provide novel compositions comprising a vesicle encapsulating an HDL mimetic peptide.
  • Another object of the present invention is to provide methods of increasing HDL levels in an animal or a human comprising administration of the pharmaceutical compositions of the present invention to the animal or the human.
  • Yet another object of the present invention is to provide methods of treating dyslipidemic or vascular disease or slowing the progression of dyslipidemic or vascular disease comprising administration of the pharmaceutical compositions of the present invention to an animal or a human in need thereof. It is yet another object of the present invention to provide new methods for the treatment of dyslipidemic or vascular disease in an animal or a human by administering the pharmaceutical compositions of the present invention.
  • the present invention provides novel compositions comprising a vesicle encapsulating an
  • HDL mimetic peptide
  • compositions of the present invention may be combined with a pharmaceutically acceptable carrier to form a pharmaceutical composition and administered to an animal or a human. These compositions cause increased lipid efflux and increase HDL levels. By increasing HDL levels, this therapeutic method treats conditions in which raising HDL levels is indicated.
  • compositions of the invention can be administered by any appropriate method as described herein, including but not limited to, oral administration or administration as a coating on a vascular stent inserted into an individual's blood vessel.
  • An individual who may be treated by administering the compositions of the present invention includes any individual who may benefit from an increase in lipid efflux or an increase in plasma HDL levels. Such individuals may have a dyslipidemic or vascular disease. Other individuals who may be treated have no clinical manifestation of disease but may be prone to such disease through genetic and/or environmental factors. Treatment of these individuals helps to delay or prevent the onset of dyslipidemic or vascular disease.
  • compositions of the present invention may be through any means described herein and includes, but is not limited to, oral administration and administration as a coating on a stent. Also described herein is a method of treating dyslipidemic and vascular disorders in an animal or a human, including administering to the animal or the human a therapeutically effective amount of the compositions of the present invention.
  • Dyslipidemic and vascular disorders amenable to treatment with the compositions disclosed herein include, but are not limited to, hyperlipidemia, hyperlipoproteinemia, hypercholesterolemia, hypertriglyceridemia, HDL deficiency, apoAl deficiency, coronary artery disease, atherosclerosis, myocardial infarction, stroke and inflammation secondary to stroke, ischemia, ischemic stroke, thrombotic stroke, peripheral vascular disease, restenosis, thrombosis, acute coronary syndrome, and reperfusion myocardial injury.
  • ABCAl ATP-binding cassette transporter
  • apoAl apolipoprotein Al
  • DMPC dimyristoyl phosphatidyl choline
  • HDL high-density lipoprotein
  • HPLC high-pressure liquid chromatography
  • LDL low-density lipoprotein
  • RBC red blood cell
  • Analog, derivative or mimetic An analog is a molecule that differs in chemical structure from a parent compound, for example a homolog (differing by an increment in the chemical structure, such as a difference in the length of an alkyl chain), a molecular fragment, a structure that differs by one or more functional groups, a change in ionization. Structural analogs are often found using quantitative structure activity relationships (QSAR), with techniques such as those disclosed in Remington (The Science and Practice of Pharmacology, 19th Edition (1995), chapter 28).
  • a derivative is a biologically active molecule derived from the base structure.
  • a mimetic is a molecule that mimics the activity of another molecule, such as a biologically active molecule.
  • Biologically active molecules can include chemical structures that mimic the biological activities of a compound.
  • Animal Living multi-cellular vertebrate organisms, a category that includes, for example, mammals and birds.
  • mammal includes both human and non-human mammals.
  • subject includes both human and veterinary subjects, for example, humans, non-human primates, dogs, cats, horses, and cows.
  • Antibody A protein (or protein complex) that includes one or more polypeptides substantially encoded by immunoglobulin genes or fragments of immunoglobulin genes.
  • the recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon, and mu constant region genes, as well as the myriad immunoglobulin variable region genes.
  • Light chains are classified as either kappa or lambda.
  • Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which in turn define the immunoglobulin classes, IgG, IgM, IgA, IgD and IgE, respectively.
  • Dyslipidemic disorder A disorder associated with any altered amount of any or all of the lipids or lipoproteins in the blood.
  • Dyslipidemic disorders include, for example, hyperlipidemia, hyperlipoproteinemia, hypercholesterolemia, hypertriglyceridemia, HDL deficiency, apoAl deficiency, and cardiovascular disease (e.g., coronary artery disease, atherosclerosis and restenosis).
  • lipid efflux refers to a process whereby lipid, such as cholesterol and phospholipid, is complexed with an acceptor, such as an apolipoprotein or apolipoprotein peptide mimetic, and removed from vesicles or cells.
  • acceptor such as an apolipoprotein or apolipoprotein peptide mimetic
  • ABCAl -dependent lipid efflux refers to a process whereby apolipoproteins or synthetic peptide mimetics of apolipoproteins bind to a cell and efflux lipid from the cell by a process that is facilitated by the ABCAl transporter.
  • Inhibiting or treating a disease Inhibiting the full development of a disease, disorder or condition, for example, in a subject who is at risk for a disease such as atherosclerosis and cardiovascular disease.
  • Treatment refers to a therapeutic intervention that ameliorates a sign or symptom of a disease or pathological condition after it has begun to develop.
  • the term “ameliorating,” with reference to a disease, pathological condition or symptom refers to any observable beneficial effect of the treatment.
  • the beneficial effect can be evidenced, for example, by a delayed onset of clinical symptoms of the disease in a susceptible subject, a reduction in severity of some or all clinical symptoms of the disease, a slower progression of the disease, a reduction in the number of relapses of the disease, an improvement in the overall health or well-being of the subject, or by other parameters well known in the art that are specific to the particular disease.
  • Isolated/purified An "isolated” or “purified” biological component (such as a nucleic acid, peptide or protein) has been substantially separated, produced apart from, or purified away from other biological components in the cell of the organism in which the component naturally occurs, that is, other chromosomal and extrachromosomal DNA and RNA, and proteins.
  • Nucleic acids, peptides and proteins that have been “isolated” thus include nucleic acids and proteins purified by standard purification methods.
  • the term also embraces nucleic acids, peptides and proteins prepared by recombinant expression in a host cell as well as chemically synthesized nucleic acids or proteins.
  • an isolated biological component is one in which the biological component is more enriched than the biological component is in its natural environment within a cell.
  • a preparation is purified such that the biological component represents at least 50%, such as at least 70%, at least 90%, at least 95%, or greater of the total biological component content of the preparation.
  • Label A detectable compound or composition that is conjugated directly or indirectly to another molecule to facilitate detection of that molecule.
  • Specific, non-limiting examples of labels include fluorescent tags, colorimetric labels, dyes, beads, enzymatic linkages, radiodense materials, and radioactive isotopes.
  • Ligand A molecule that can bind to another molecule.
  • Linker A molecule that joins two other molecules, either covalently, or through ionic, van der Waals or hydrogen bonds.
  • Lipid A class of water-insoluble, or partially water insoluble, oily or greasy organic substances, that are extractable from cells and tissues by nonpolar solvents, such as chloroform or ether.
  • Types of lipids include triglycerides (e.g., natural fats and oils composed of glycerin and fatty acid chains), glycolipids, phospholipids (e.g., phosphatidylethanolamine, phosphatidylcholine, phosphatidylserine, and phosphatidylinositol), sphingolipids (e.g., sphingomyelin, cerebrosides and gangliosides), and sterols (e.g., cholesterol).
  • triglycerides e.g., natural fats and oils composed of glycerin and fatty acid chains
  • glycolipids e.g., phospholipids (e.g., phosphatidylethanolamine, phosphatidylcholine, phosphatidylserine, and phosphatidylinositol)
  • sphingolipids e.g., sphingomyelin
  • Non-cytotoxic is one that does not substantially affect the viability or growth characteristics of a cell at a dosage normally used to treat the cell or a subject. Furthermore, the percentage of cells releasing intracellular contents, such as LDH or hemoglobin, is low (e.g., about 10% or less) in cells treated with a non-cytotoxic compound. Lipid efflux from a cell that occurs by a non-cytotoxic compound results in the removal of lipid from a cell by a process that maintains the overall integrity of the cell membrane and does not lead to significant cell toxicity.
  • Non-polar A non-polar compound is one that does not have concentrations of positive or negative electric charge.
  • Non-polar compounds such as, for example, oil, are not well soluble in water.
  • Peptide A polymer in which the monomers are amino acid residues which are joined together through amide bonds.
  • the amino acids are alpha-amino acids, either the L-optical isomer or the D-optical isomer can be used.
  • the amino acid sequences disclosed herein are shown using standard single letter abbreviations as known to one of ordinary skill in the art, and in some cases as three letter codes for amino acids, as defined in 37 C.F.R. 1.822 and as commonly known to one of ordinary skill in the art.
  • the single letter designation for an amino acid for example S for serine is shown in lower case, s, the serine is a D amino acid.
  • peptide or “polypeptide” as used herein are intended to encompass any amino acid sequence and include modified sequences such as glycoproteins.
  • peptide is specifically intended to cover naturally occurring peptides, as well as those which are recombinantly or synthetically produced.
  • residue or “amino acid residue” includes reference to an amino acid that is incorporated into a peptide, polypeptide, or protein.
  • the peptides presented herein are read from the N to the C terminus i.e., from left to right. Accordingly, the N terminal amino acid in Leu GIu Ser is Leu and the C- terminal amino acid is Ser.
  • Peptides of the present invention include peptides with substitutions for amino acids in the peptide sequence. Such substitutions may be conservative substitutions, isosteric substitutions, substitutions between isosteric groups, and non-conservative substitutions as defined herein.
  • Peptides of the present invention include conservatively substituted peptides, wherein these conservative substitutions occur at 1%, 3%, 5%, 7%, 10%, 15%, 20%, 25%, 30%, 40%, or 50% of the amino acid residues.
  • Peptides of the present invention include peptides that are homologous at 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99% of the entire sequence of the peptide.
  • compositions and formulations suitable for pharmaceutical delivery of one or more therapeutic compounds or molecules, and additional pharmaceutical agents are conventional.
  • Remington 's Pharmaceutical Sciences, by E. W. Martin, Mack Publishing Co., Easton, PA, 15th Edition (1975) describes compositions and formulations suitable for pharmaceutical delivery of one or more therapeutic compounds or molecules, and additional pharmaceutical agents.
  • parenteral formulations usually comprise injectable fluids that include pharmaceutically and physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or the like as a vehicle.
  • pharmaceutically and physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or the like as a vehicle.
  • physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or the like
  • solid compositions e.g., powder, pill, tablet, or capsule forms
  • conventional non-toxic solid carriers can include, for example, pharmaceutical grades of mannitol, lactose, starch, or magnesium stearate.
  • compositions to be administered can contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, preservatives, and pH buffering agents and the like, for example sodium acetate or sorbitan monolaurate.
  • non-toxic auxiliary substances such as wetting or emulsifying agents, preservatives, and pH buffering agents and the like, for example sodium acetate or sorbitan monolaurate.
  • a phospholipid consists of a water-soluble polar head, linked to two water- insoluble non-polar tails (by a negatively charged phosphate group). Both tails consist of a fatty acid, each about 14 to about 24 carbon groups long. When placed in an aqueous environment, phospholipids form a bilayer or micelle, where the hydrophobic tails line up against each other. This forms a membrane with hydrophilic heads on both sides.
  • a phospholipid is a lipid that is a primary component of animal cell membranes.
  • Polar A polar molecule is one in which the centers of positive and negative charge distribution do not converge. Polar molecules are characterized by a dipole moment, which measures their polarity, and are soluble in other polar compounds and virtually insoluble in nonpolar compounds.
  • Recombinant nucleic acid A sequence that is not naturally occurring or has a sequence that is made by an artificial combination of two otherwise separated segments of sequence. This artificial combination is often accomplished by chemical synthesis or, more commonly, by the artificial manipulation of isolated segments of nucleic acids, for example, by genetic engineering techniques such as those described in Sambrook et al. (ed.), Molecular Cloning: A Laboratory Manual, 2 nd ed., vol. 1-3, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989. The term recombinant includes nucleic acids that have been altered solely by addition, substitution, or deletion of a portion of the nucleic acid. Therapeutically effective amount: A quantity of a specified agent sufficient to achieve a desired effect in a subject being treated with that agent.
  • this can be the amount of a composition useful in preventing, ameliorating, and/or treating a dyslipidemic disorder ⁇ e.g., atherosclerosis) in a subject.
  • a therapeutically effective amount of an agent is an amount sufficient to prevent, ameliorate, and/or treat a dyslipidemic disorder (e.g., atherosclerosis) in a subject without causing a substantial cytotoxic effect (e.g., membrane microsolubilization) in the subject.
  • the effective amount of an agent useful for preventing, ameliorating, and/or treating a dyslipidemic disorder (e.g., atherosclerosis) in a subject will be dependent on the subject being treated, the severity of the disorder, and the manner of administration of the therapeutic composition.
  • a "transformed" cell is a cell into which has been introduced a nucleic acid molecule by molecular biology techniques.
  • the term encompasses all techniques by which a nucleic acid molecule might be introduced into such a cell, including transfection with viral vectors, transformation with plasmid vectors, and introduction of naked DNA by electroporation, lipofection, and particle gun acceleration.
  • compositions of the present invention comprise a vesicle encapsulating an HDL mimetic peptide. These compositions may be combined with a pharmaceutically acceptable carrier to make a pharmaceutical composition for administration to an animal or a human.
  • a pharmaceutically acceptable carrier to make a pharmaceutical composition for administration to an animal or a human.
  • compositions comprise an HDL mimetic peptide encapsulated within a vesicle.
  • vesicle includes vesicles of various sizes, including but not limited to nanoparticles. Thus, as used herein, the term “vesicle” also refers to nanoparticles.
  • the vesicles of the present invention include, but are not limited to, lipid-containing vesicles (e.g., liposomes).
  • the vesicle can be a lipid vesicle as described in US Patent No. 5,985,852.
  • lipid vesicles containing peptides are well-known in the art, e.g., sonication methods, phase inversion methods, and oil/water emulsion methods.
  • the lipid vesicles containing peptides can be prepared as described in US Patent No. 5,916,588.
  • a stock solution of a peptide to be encapsulated is prepared at a concentration of 3 mg/ml in 50 mM NaPO 4 buffer.
  • a stock solution of a lipid such as dipalmityl phosphatidylcholine (DPPC) is prepared at a concentration of 20 mg/ml in chloroform:methanol (3:1).
  • the peptide is combined with the lipid at a molar recombination ratio range of peptide to lipid from 1 :5 up to 1 :300.
  • Appropriate aliquots of peptide and lipid are mixed and the sample is incubated at 50° C for about 30 minutes to about 3 hrs. to obtain clarity of the solution. Up to three cycles of heating and cooling may be required to obtain clarity of the sample.
  • the sample is filtered over a polyethersulfone (PES) filter to remove unincorporated lipid.
  • Lipids for incorporation into vesicles can be any suitable lipid or mixture of lipids, including, but not limited to, the electrically-neutral and electrically-charged phospholipids described hereinbelow.
  • Nanoparticles include, but are not limited to, those described in US Patent Application Serial No. 10/362,848 (US 2004/0063606), which describes biodegradable polymers, such as polyester amide polymers, polyester urethane polymers, and polyester urea polymers, that contain at least one amino acid and a non-amino acid moiety per repeat unit.
  • biodegradable polymers such as polyester amide polymers, polyester urethane polymers, and polyester urea polymers, that contain at least one amino acid and a non-amino acid moiety per repeat unit.
  • Such vesicles and nanoparticles are well-known in the art and the present peptides are encapsulated into such nanoparticles according to well-known methods, e.g., as described in the foregoing publications.
  • Hydrogel-type nanoparticles as described, e.g., in US 2006/0177416 (see, e.g., page 1, paragraph 0006), can also be used in the compositions of the invention, for example, for compositions intended for oral administration.
  • Other examples of nanoparticles suitable for oral administration include but are not limited to the nanoparticles described in Damge C. et al., Expert Opinion on Drug Delivery 5:45-68 (2008); Galindo-Rodriguez S. A., Critical Reviews in Therapeutic Drug Carrier Systems, 22:419-464 (2005); Dong Y. and Feng S.-S., Biomaterials 26:6068-6076 (2005); and Pamajula S. et al., J. Pharm. Pharmacol.
  • nanoparticles suitable for oral administration are well-known in the art and the present peptides are encapsulated into such nanoparticles according to well-known methods, e.g., as described in the foregoing publications.
  • PLGA nanoparticles are well-known in the art and the present peptides are encapsulated into such nanoparticles according to well-known methods, e.g., as described in Nam SE, et al, supra.
  • US Patent No. 7,037,552 Coating Dispensing System and Method Using a Solenoid Head for Coating Medical Devices
  • a therapeutic agent including a biological molecule, such as a protein
  • US Patent Publication No. 2004/0170685 Bioactive Stents and Methods for Use Thereof
  • US Patent Publication No. 2004/0170685 also describes methods for producing biodegradable stents to which peptides are bound (see, e.g., paragraphs 0035-0040).
  • nanoparticles for use in the methods of the invention include those described in U.S. Patent Application Serial Nos. 11/344,689 (US 2006/0177416), 11/345,021 (US 2006/0188469), 11/345,815 (US 2006/0188486), and related applications.
  • the nanoparticles are chosen from those recited in the claims of US Patent Application Serial No. 10/362,848 (US 2004/0063606).
  • Some appropriate nanoparticles for use in the compositions of the invention are disclosed in Reddy et al., Clin. Cancer Res. (2006) 12(22), 6677-6686.
  • Peptides for use in the compositions of the invention are encapsulated into vesicles or nanoparticles using methods that are well-known in the art, e.g. but not limited to any of the methods set forth in the foregoing publications.
  • the encapsulated peptides of the invention can be used for the manufacture of medicaments for use in increasing lipid efflux, increasing HDL levels, and/or treating a dyslipidemic or vascular disorder in an individual receiving the medicament.
  • Medicaments containing peptides of the invention can be manufactured for oral administration, for coating a vascular stent, for intravenous or intramuscular administration, or for administration by any other means known in the art and/or set forth herein.
  • Administration of the encapsulated peptides to individuals in need thereof can be through any appropriate means, including but not limited to oral administration or as a coating on a stent.
  • the HDL mimetic peptides for use in the compositions present invention include the peptides set forth hereinbelow and as incorporated by reference. These applications incorporated by reference in their entirety include USSN 60/898,132, filed January 29, 2007, 11/764,619 filed June 18, 2007, 11/861,758 filed September 26, 2007, 61/023,963 filed January 28, 2008, 12/021,486 filed January 29, 2008, and 12/021,629 filed January 29, 2008. HDL mimetic peptides mimic naturally occuring ApoAI by binding lipid and by facilitating lipid efflux from cells. Peptides in the present compositions can be unmodified or modified as described herein and/or as will be appreciated by one of ordinary skill in the art.
  • amino- and/or carboxy-termini of the peptides of the invention can be chemically modified by conjugation with various functional groups.
  • synthetic peptides carry free amino- and carboxy-termini, which carry, respectively, a positive and a negative charge.
  • amino-terminal acetylation and/or carboxy-terminal amidation neutralizes, respectively, the amino-terminal and/or carboxy-terminal charge, such that the synthetic peptide more closely mimics the structure and function of the natural peptide.
  • Such amino- and carboxy- terminal modification also renders peptides more resistant to enzymatic degradation, such as that resulting from exopeptidases.
  • Neutralization of the terminal charge of synthetic peptide mimetics of apolipoproteins has been shown to increase their lipid affinity (Yancey et al., Biochem. 34:7955- 7965, 1995; Venkatachalapathi et al., Protein: Structure, Function and Genetics 15:349-359, 1993).
  • acetylation of the amino-terminus of amphipathic peptides increases the lipid affinity of the peptide (Mishra et al., J. Biol. Chem. 269:7185-7191, 1994).
  • any of the peptides employed in compositions of the present invention may contain a D amino acid at the N-terminus, at the C- terminus, or at both the N- and C-termini.
  • the presence of these D amino acids can help protect against peptide degradation, hi some embodiments, all the amino acids of any peptide of the present invention may be D amino acids. This embodiment is useful for protection against degradation following oral administration of a pharmaceutical composition comprising the peptides of the present invention.
  • Some examples of peptides of the present invention containing one or more D amino acids include but are not limited to those shown hereinbelow. D amino acids are indicated in all-upper-case letters, e.g., "SER" or "PRO.
  • the peptides for use in the present compositions can be modified by any other chemical modification that protects the present peptides from exopeptidase or endopeptidase digestion.
  • the present peptides can be modified by a detectable moiety, as described herein and/or as will be appreciated by the skilled artisan.
  • any of the peptides used in compositions of the present invention can contain one or more amino acid substitutions, e.g., an isosteric amino acid substitution, a conservative amino acid substitution, and/or a non-conservative amino acid substitution as described hereinbelow.
  • SEQ ID NO: 21 Ac-Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn;
  • SEQ ID NO: 22 Ac-Ser Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn;
  • SEQ ID NO: 26 Ac-Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp; SEQ ID NO: 27 Ac-Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn;
  • SEQ ID NO: 28 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp Trp;
  • SEQ ID NO: 46 Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Tip Arg Trp Trp Trp Trp-NH 2 ; SEQ ID NO: 47 Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GkI-NH 2 ;
  • SEQ ID NO: 65 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn-NH 2 ; SEQ ID NO: 66 Ac-Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Trp Trp Trp Trp Trp Trp Trp Trp-NH 2 ;
  • SEQ ID NO: 68 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp Trp Trp Trp Trp-NH 2 ; SEQ ID NO: 69 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala
  • SEQ ID NO: 70 Ac-Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr
  • SEQ ID NO: 78 Ac-Asp Tip Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe Pro Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala AIa-NH 2 ;
  • SEQ ID NO: 102 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn; SEQ ID NO: 103 Ac-SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp Trp;
  • SEQ ID NO: 104 Ac-SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn;
  • SEQ ID NO: 105 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp Trp Trp; SEQ ID NO: 106 Ac-PRO Arg
  • Trp Trp Trp Trp-NH 2 SEQ ID NO: 119 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn-NH 2 ; SEQ ID NO: 120 SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Trp Trp Trp Trp Trp Trp Trp Trp-NH
  • SEQ ID NO: 150 ALA Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala Phe;
  • SEQ ID NO: 160 Ac-PRO VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys GIn Lys Leu LyS-NH 2 .
  • SEQ ID NO: 162 Ser Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 163 Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS;
  • SEQ ID NO: 180 Ac-Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn
  • SEQ ID NO: 200 Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP-NH 2 ;
  • SEQ ID NO: 201 Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH 2 ;
  • SEQ ID NO: 213 Ac-Ser Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH 2 ;
  • SEQ ID NO: 214 Ac-Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS-NH 2 ;
  • SEQ ID NO: 224 Ac-Leu GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys Pro Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu LYS-NH 2 ;
  • SEQ ID NO: 225 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS- NH 2 ;
  • SEQ ID NO: 226 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS-NH 2 ;
  • SEQ ID NO: 227 Ac-Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS- NH 2 ;
  • SEQ ID NO: 228 Ac-Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS-NH 2 ; SEQ ID NO: 229 Asp Trp Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe Pro Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala ALA; SEQ ID NO: 230 Ala Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala PHE;
  • SEQ ID NO: 231 Pro VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys GIn Lys Leu LYS;
  • SEQ ID NO: 232 Ac-Asp Trp Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe Pro Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala ALA; SEQ ID NO: 233 Ac-Ala Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala PHE;
  • SEQ ID NO: 240 Ac-Pro VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys GIn Lys Leu LYS-NH 2 .
  • SEQ ID NO: 242 SER Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 243 SER Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS;
  • SEQ ID NO: 244 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp TRP; SEQ ID NO: 245 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr
  • SEQ ID NO: 248 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp TRP;
  • SEQ ID NO: 249 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 250 SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Trp Trp Trp TRP;
  • SEQ ID NO: 258 Ac-SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 259 Ac-SER Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN;
  • SEQ ID NO: 264 Ac-SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 265 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Trp Trp Trp Trp TRP;
  • SEQ ID NO: 266 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN;
  • SEQ ID NO: 279 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH 2 ; SEQ ID NO: 280 SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Trp Trp Trp Trp Trp TRP-NH 2 ;
  • Trp TRP-NH 2 SEQ ID NO: 296 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met
  • Trp TRP-NH 2 Trp TRP-NH 2 ;
  • ARGTRPTRPTRPTRP-NH 2 ARGTRPTRPTRPTRP-NH 2 ;
  • GLU ALA PHE-NH 2 SEQ ID NO: 397 PRO VAL LEU ASP LEU PHE ARG GLU LEU LEU ASN GLU LEU LEU
  • the HDL mimetic peptides of the present invention may be complexed with one or more lipids, e.g., one or more electrically neutral and/or electrically charged phospholipids. Particularly for oral administration, these peptide/lipid complexes may then be encapsulated into vesicles or nanoparticles as described herein. In one example, peptides are reconstituted with one or more lipids prior to encapsulating the peptide/lipid complexes within vesicles or nanoparticles.
  • the peptides may be reconstituted with an electrically neutral phospholipid (i.e., a phospholipid having zero net electrical charge), e.g., a lipid containing phospatidylcholine (lecithin) or sphingomyelin or a mixture thereof.
  • an electrically neutral phospholipid i.e., a phospholipid having zero net electrical charge
  • a lipid containing phospatidylcholine (lecithin) or sphingomyelin a mixture thereof.
  • the peptides may also be reconstituted with an electrically charged phospholipid, e.g., but not limited to, a lipid containing phosphatidylserine, phosphatidylinositol, phosphatidylglycerol, l,2-diacylglycerol-3-phosphate (phosphatidic acid), and/or a mixture thereof (each of these phospholipids has a net electrical charge of minus one).
  • the peptide is reconstituted with a neutral phospholipid plus a charged phospholipid.
  • Such charged lipoprotein complexes containing an HDL mimetic peptide or any full-length apoplipoprotein such as but not limited to ApoA-I can be produced using well-known methods, e.g. but not limited to the methods set forth in US 20060217312 or US 7,189,411.
  • Such HDL mimetic peptide/lipid complexes or apolipoprotein/lipid complexes can then be encapsulated into vesicles or nanoparticles as described herein.
  • neutral phospholipids containing phosphatidylcholine that may be employed to produce peptide/lipid complexes include but are not limited to the following: dipalmitoylphosphatidylcholine (DPPC); dioleoylphosphatidylcholine (DOPC); l-palmitoyl-2- oleoylphosphatidylcholine (POPC); l-palmitoyl-2-linoleoylphosphatidylcholine (PLPC); 1- palmitoyl-2-arachidonylphosphatidylcholine (PAPC); l-palmitoyl-2- docosahexanoylphosphatidylcholine (PDPC); and l-palmitoyl-2-myristoylphosphatidylcholine (PMPC).
  • DPPC dipalmitoylphosphatidylcholine
  • DOPC dioleoylphosphatidylcholine
  • POPC l-palmitoy
  • the peptides of the present invention may be complexed with neutral and/or charged phospholipids or any mixture thereof in molar ratios ranging from about 1:0.5 to about 1:200.
  • the peptide to lipid ratio can be, but is not limited to, 1:0.5 to 1:10; 1:1 to 1: 5; 1:2 to 1:10; 1 : 1 to 1 :20; 1 :40 to 1 : 100; 1 : 100.1 to 1 :200. Any ratio within these ranges may be employed.
  • the molar ratio of charged phospholipid to neutral phospholipid may be (but is not limited to), e.g., about 400:1 to about 1:400 (e.g., about 1:1 to 1:10, 1:5 to 1:20, 1:5 to 1:50, 1:10 to 1:100, 1:20 to 1:200, 1:40 to 1:400, 1:1 to 10:1, 5:1 to 20:1, 5:1 to 50:1, 10:1 to 100:1, 20:1 to 200:1, 40:1 to 400:1). Any ratio within these ranges may be employed in the present compositions. See, e.g., US 20060217312 for other appropriate ratios of peptide to phospholipid and charged phospholipid to uncharged phospholipid.
  • the present invention may be used for the production of the peptides or peptide analogs of the present invention.
  • Proteins typically L-amino acids
  • oligopeptides are chains of amino acids (typically L-amino acids) whose alpha carbons are linked through peptide bonds formed by a condensation reaction between the carboxyl group of the alpha carbon of one amino acid and the amino group of the alpha carbon of another amino acid.
  • the terminal amino acid at one end of the chain i.e., the amino terminal
  • the terminal amino acid at the other end of the chain i.e., the carboxy terminal
  • amino terminus refers to the free alpha-amino group on the amino acid at the amino terminal of the protein, or to the alpha-amino group (imino group when participating in a peptide bond) of an amino acid at any other location within the protein.
  • carboxy terminus refers to the free carboxyl group on the amino acid at the carboxy terminus of a protein, or to the carboxyl group of an amino acid at any other location within the protein.
  • amino acids making up a protein are numbered in order, starting at the amino terminal and increasing in the direction toward the carboxy terminal of the protein. Thus, when one amino acid is said to "follow" another, that amino acid is positioned closer to the carboxy terminal of the protein than the preceding amino acid.
  • residue is used herein to refer to an amino acid (D or L) or an amino acid mimetic that is incorporated into a protein by an amide bond.
  • D amino acid amino acid
  • amino acid mimetic amino acid mimetic that is incorporated into a protein by an amide bond.
  • amino acid may be a naturally occurring amino acid or, unless otherwise limited, may encompass known analogs of natural amino acids that function in a manner similar to the naturally occurring amino acids (i.e., amino acid mimetics).
  • amino acid mimetics i.e., amino acid mimetics
  • an amide bond mimetic includes peptide backbone modifications well known to those skilled in the art.
  • the present invention includes HDL mimetic peptides containing isosteric amino acid substitutions at specific amino acid positions within the peptides.
  • isosteric substitution is meant that an amino acid at a particular position within a peptide of the invention can be substituted with another amino acid belonging to the same isosteric group as described hereinbelow.
  • Amino acids within a given isosteric group, as set forth hereinbelow, are amino acids having similar size, shape, polar/nonpolar properties, charge, and/or steric properties.
  • the invention provides peptides wherein substitution of an amino acid with an amino acid belonging to the same isosteric group allows the substituted peptide to retain at least about 20% of the biological activity of the unsubstituted peptide, e.g., at least about: 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 110%, 125%, 150%, 175%, 200%, 250%, 300%, or more, of the biological activity of the unsubstituted peptide.
  • biological activity of the peptide is meant the ability of the peptide to promote lipid efflux and and/or have an anti-inflammatory effect, as described hereinbelow.
  • the invention also provides peptides having an amino acid substituted with an amino acid belonging to a different isosteric group from the original amino acid, such that the substitution allows the peptide to retain at least about 20% of the biological activity (e.g., lipid efflux and/or anti- inflammatory properties) of the unsubstituted peptide, e.g., at least about: 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 110%, 125%, 150%, 175%, 200%, 250%, 300%, or more, of the biological activity of the unsubstituted peptide.
  • the biological activity e.g., lipid efflux and/or anti- inflammatory properties
  • Isosteric amino acid groupings of the present invention are as follows: a) Isosteric Group 1 : Lys, His, and Arg; b) Isosteric Group 2: Asp and GIu; c) Isosteric Group 3: Ser, Thr, Leu, He, GIy, Ala, VaI, and GABA; d) Isosteric Group 4: Phe and Tyr; and e) Isosteric Group 5: Pro.
  • An amino acid belonging to a given isosteric group can be substituted by a different amino acid within that same isosteric group.
  • an amino acid within a given isosteric group can be substituted by an amino acid from a different isosteric group as follows: a) an amino acid within Isosteric Group 1 can be substituted with an amino acid within
  • Isosteric Group 2 b) an amino acid within Isosteric Group 2 can be substituted with an amino acid within Isosteric Group 1, Isosteric Group 3, or Isosteric Group 4; c) an amino acid within Isosteric Group 3 can be substituted with an amino acid within Isosteric Group 4; moreover, an amino acid within Isosteric Group 3 that resides on a hydrophilic surface of an amphipathic helix of an HDL mimetic peptide of the invention can be substituted with an amino acid within either Isosteric Group 1 or Isosteric Group 2; d) An amino acid within Isosteric Group 4 can be substituted with an amino acid within Isosteric Group 3.
  • I Isoleucine
  • L Leucine
  • M Methionine
  • V Valine
  • F Phenylalanine
  • Y Tyrosine
  • W Tryptophan
  • a conservative substitution is a substitution in which the substituting amino acid (naturally occurring or modified) is structurally related to the amino acid being substituted, i.e., has about the same size and electronic properties as the amino acid being substituted. Thus, the substituting amino acid would have the same or a similar functional group in the side chain as the original amino acid.
  • a “conservative substitution” also refers to utilizing a substituting amino acid which is identical to the amino acid being substituted except that a functional group in the side chain is protected with a suitable protecting group.
  • Peptides of the present invention include conservatively substituted peptides, wherein these conservative substitutions occur at 1%, 3%, 5%, 7%, 10%, 15%, 20%, 25%, 30%, 40%, or 50% of the amino acid residues.
  • Peptides of the present invention include peptides that are homologous at 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99% of the entire sequence of the peptide.
  • Suitable protecting groups are described in Green and Wuts, "Protecting Groups in Organic Synthesis", John Wiley and Sons, Chapters 5 and 7, 1991, the teachings of which are incorporated herein by reference.
  • Preferred protecting groups are those which facilitate transport of the peptide through membranes, for example, by reducing the hydrophilicity and increasing the lipophilicity of the peptide, and which can be cleaved, either by hydrolysis or enzymatically (Ditter et al., 1968. J. Pharm. Sci. 57:783; Ditter et al., 1968. J. Pharm. Sci. 57:828; Ditter et al., 1969. J. Pharm. Sci.
  • Suitable hydroxyl protecting groups include ester, carbonate and carbamate protecting groups.
  • Suitable amine protecting groups include acyl groups and alkoxy or aryloxy carbonyl groups, as described above for N-terminal protecting groups.
  • Suitable carboxylic acid protecting groups include aliphatic, benzyl and aryl esters, as described below for C-terminal protecting groups.
  • the carboxylic acid group in the side chain of one or more glutamic acid or aspartic acid residues in a peptide of the present invention is protected, preferably as a methyl, ethyl, benzyl or substituted benzyl ester, more preferably as a benzyl ester.
  • Group I includes leucine, isoleucine, valine, methionine and modified amino acids having the following side chains: ethyl, n-propyl n-butyl.
  • Group I includes leucine, isoleucine, valine and methionine.
  • Group II includes glycine, alanine, valine and a modified amino acid having an ethyl side chain.
  • Group II includes glycine and alanine.
  • Group III includes phenylalanine, phenylglycine, tyrosine, tryptophan, cyclohexylmethyl glycine, and modified amino residues having substituted benzyl or phenyl side chains.
  • Preferred substituents include one or more of the following: halogen, methyl, ethyl, nitro, — NH 2 , methoxy, ethoxy and — CN.
  • Group III includes phenylalanine, tyrosine and tryptophan.
  • Group IV includes glutamic acid, aspartic acid, a substituted or unsubstituted aliphatic, aromatic or benzylic ester of glutamic or aspartic acid (e.g., methyl, ethyl, n-propyl iso-propyl, cyclohexyl, benzyl or substituted benzyl), glutamine, asparagine, — CO — NH — alkylated glutamine or asparagines (e.g., methyl, ethyl, n-propyl and iso-propyl) and modified amino acids having the side chain — (CH 2 ) 3 — COOH, an ester thereof (substituted or unsubstituted aliphatic, aromatic or benzylic ester), an amide thereof and a substituted or unsubstituted N-alkylated amide thereof.
  • glutamic acid e.g., methyl, ethyl, n-prop
  • Group IV includes glutamic acid, aspartic acid, methyl aspartate, ethyl aspartate, benzyl aspartate and methyl glutamate, ethyl glutamate and benzyl glutamate, glutamine and asparagine.
  • Group V includes histidine, lysine, ornithine, arginine, N-nitroarginine, ⁇ -cycloarginine, ⁇ - hydroxyarginine, N-amidinocitruline and 2-amino-4-guanidinobutanoic acid, homologs of lysine, homologs of arginine and homologs of ornithine.
  • Group V includes histidine, lysine, arginine and ornithine.
  • a homolog of an amino acid includes from 1 to about 3 additional or subtracted methylene units in the side chain.
  • Group VI includes serine, threonine, and modified amino acids having C1-C5 straight or branched alkyl side chains substituted with — OH or — SH, for example, — CH 2 CH 2 OH, -CH 2 CH 2 CH 2 OH or -CH 2 CH 2 OHCH 3 .
  • Group VI includes serine, or threonine.
  • suitable substitutions for amino acid residues include “severe” or “non- conservative” substitutions.
  • the terms “severe” or “non-conservative” substitution are used interchangeablt throughout this application.
  • a “severe substitution” is a substitution in which the substituting amino acid (naturally occurring or modified) has significantly different size and/or electronic properties compared with the amino acid being substituted.
  • the side chain of the substituting amino acid can be significantly larger (or smaller) than the side chain of the amino acid being substituted and/or can have functional groups with significantly different electronic properties than the amino acid being substituted.
  • severe substitutions of this type include the substitution of phenylalanine or cyclohexylmethyl glycine for alanine, isoleucine for glycine, a D amino acid for the corresponding L amino acid, or — NH — CH[( — CH 2 ) 5 — COOH] — CO — for aspartic acid.
  • a functional group may be added to the side chain, deleted from the side chain or exchanged with another functional group.
  • Examples of severe substitutions of this type include adding of valine, leucine or isoleucine, exchanging the carboxylic acid in the side chain of aspartic acid or glutamic acid with an amine, or deleting the amine group in the side chain of lysine or ornithine, hi yet another alternative, the side chain of the substituting amino acid can have significantly different steric and electronic properties that the functional group of the amino acid being substituted. Examples of such modifications include tryptophan for glycine, lysine for aspartic acid and — (CH 2 ) 4 COOH for the side chain of serine. These examples are not meant to be limiting.
  • amino acid residues in the peptides may be substituted with naturally occurring non-encoded amino acids and synthetic amino acids.
  • Certain commonly encountered amino acids which provide useful substitutions include, but are not limited to, ⁇ -alanine and other omega-amino acids, such as 3- aminopropionic acid, 2,3-diaminopropionic acid, 4-aminobutyric acid and the like; ⁇ - aminoisobutyric acid; ⁇ -aminohexanoic acid; ⁇ -aminovaleric acid; N-methylglycine or sarcosine; ornithine; citrulline; t-butylalanine; t-butylglycine; N-methylisoleucine; phenylglycine; cyclohexylalanine; norleucine; naphthylalanine; 4-chlorophenylalanine; 2-fluorophenylalanine; 3- fluorophenylalanine; 4-
  • the amino acids of the peptides will be substituted with L- amino acids, the substitutions are not limited to L-amino acids.
  • modified forms of the peptides wherein an L-amino acid is replaced with an identical D-amino acid (e.g., L-Arg— >D-Arg) or with a conservatively-substituted D-amino acid (e.g., LArg— >D-Lys), and vice versa.
  • mimetic compounds are synthetic compounds having a three-dimensional structure (of at least part of the mimetic compound) that mimics, for example, the primary, secondary, and/or tertiary structural, and/or electrochemical characteristics of a selected peptide, structural domain, active site, or binding region (e.g., a homotypic or heterotypic binding site, a catalytic active site or domain, a receptor or ligand binding interface or domain, or a structural motif) thereof.
  • a homotypic or heterotypic binding site e.g., a homotypic or heterotypic binding site, a catalytic active site or domain, a receptor or ligand binding interface or domain, or a structural motif
  • the mimetic compound will often share a desired biological activity with a native peptide, as discussed herein (e.g., the ability to interact with lipids).
  • a desired biological activity e.g., the ability to interact with lipids.
  • at least one subject biological activity of the mimetic compound is not substantially reduced in comparison to, and is often the same as or greater than, the activity of the native peptide on which the mimetic was modeled.
  • mimetic compounds of the disclosure can have other desired characteristics that enhance their therapeutic application, such as increased cell permeability, greater affinity and/or avidity for a binding partner, and/or prolonged biological half-life.
  • the mimetic compounds of the disclosure can have a backbone that is partially or completely non- peptide, but with side groups identical to the side groups of the amino acid residues that occur in the peptide on which the mimetic compound is modeled.
  • Several types of chemical bonds for example, ester, thioester, thioamide, retroamide, reduced carbonyl, dimethylene and ketomethylene bonds, are known in the art to be generally useful substitutes for peptide bonds in the construction of protease-resistant mimetic compounds.
  • peptides useful within the disclosure are modified to produce synthetic peptide mimetics by replacement of one or more naturally occurring side chains of the 20 genetically encoded amino acids (or D-amino acids) with other side chains, for example with groups such as alkyl, lower alkyl, cyclic 4-, 5-, 6-, to 7-membered alkyl, amide, amide lower alkyl, amide di(lower alkyl), lower alkoxy, hydroxy, carboxy and the lower ester derivatives thereof, and with 4-, 5-, 6-, to 7-membered heterocyclics.
  • groups such as alkyl, lower alkyl, cyclic 4-, 5-, 6-, to 7-membered alkyl, amide, amide lower alkyl, amide di(lower alkyl), lower alkoxy, hydroxy, carboxy and the lower ester derivatives thereof, and with 4-, 5-, 6-, to 7-membered heterocyclics.
  • proline analogs can be made in which the ring size of the proline residue is changed from a 5-membered ring to a 4-, 6-, or 7-membered ring.
  • Cyclic groups can be saturated or unsaturated, and if unsaturated, can be aromatic or non-aromatic.
  • Heterocyclic groups can contain one or more nitrogen, oxygen, and/or sulphur heteroatoms.
  • Examples of such groups include furazanyl, furyl, imidazolidinyl, imidazolyl, imidazolinyl, isothiazolyl, isoxazolyl, morpholinyl (e.g., morpholino), oxazolyl, piperazinyl (e.g., 1-piperazinyl), piperidyl (e.g., 1-piperidyl, piperidino), pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridyl, pyrinidinyl, pyrrolidinyl (e.g., 1- pyrrolidinyl), pyrrolinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, thiomorpholinyl (e.g., thiomorpholino), and thiazolyl groups.
  • heterocyclic groups can be substituted or unsubstituted.
  • the substituent can be alkyl, alkoxy, halogen, oxygen, or substituted or unsubstituted phenyl.
  • Peptides, as well as peptide analogs and mimetics can also be covalently bound to one or more of a variety of nonproteinaceous polymers, for example, polyethylene glycol, polypropylene glycol, or polyoxyalkenes, as described in U.S. Pat. Nos. 4,640,835; 4,496,689; 4,301,144; 4,670,417; 4,791,192; and 4,179,337.
  • peptide analogs and mimetics within the scope of the disclosure include glycosylation variants, and covalent or aggregate conjugates with other chemical moieties.
  • Covalent derivatives can be prepared by linkage of functionalities to groups which are found in amino acid side chains or at the N- or C-termini, by means which are well known in the art. These derivatives can include, without limitation, aliphatic esters or amides of the carboxyl terminus, or of residues containing carboxyl side chains, O-acyl derivatives of hydroxyl group- containing residues, and N-acyl derivatives of the amino terminal amino acid or amino-group containing residues (e.g., lysine or arginine).
  • Acyl groups are selected from the group of alky 1- moieties including C3 to Cl 8 alkyl, thereby forming alkanoyl aroyl species. Also embraced are versions of a native primary amino acid sequence which have other minor modifications, including phosphorylated amino acid residues, for example, phosphotyrosine, phosphoserine, or phosphothreonine, or other moieties, including ribosyl groups or cross-linking reagents.
  • the linkage between amino acid residues can be a peptide bond or amide linkage (e.g., -C-C(O)NH-).
  • amide linkages e.g., -C-C(O)NH-.
  • one or more amide linkages is optionally replaced with a linkage other than amide, for example, a substituted amide.
  • Substituted amides generally include, but are not limited to, groups of the formula -C(O)NR-, where R is (Cj-C 6 ) alkyl, substituted (Ci-C 6 ) alkyl, (C 1 -C 6 ) alkenyl, substituted (C 1 -C 6 ) alkenyl, (Ci-C 6 ) alkynyl, substituted (C 1 -C 6 ) alkynyl, (C 5 -C 20 ) aryl, substituted (C 5 -C 20 ) aryl, (C 6 -C 26 ) alkaryl, substituted (C 6 -C 26 ) alkaryl, 5-20 membered heteroaryl, substituted 5-20 membered heteroaryl, 6-26 membered alkheteroaryl, and substituted 6-26 membered alkheteroaryl.
  • R is (Cj-C 6 ) alkyl, substituted (Ci-C 6 ) alkyl
  • one or more amide linkages can be replaced with peptidomimetic or amide mimetic moieties which do not significantly interfere with the structure or activity of the peptides. Suitable amide mimetic moieties are described, for example, in Olson et ah, J. Med. Chem. 36:3039-3049, 1993.
  • the peptides of the present invention may optionally be acetylated at the N-terminus.
  • the peptides of the present invention may optionally have a carboxy terminal amide.
  • the peptides of the present invention may have both an acetylated N-terminus and a carboxy terminal amide. Methods of acetylating the N-terminus or adding a carboxy terminal amide are well known to one of ordinary skill in the art. E. Labeled Peptides
  • a detectable moiety can be linked to any of the compositions disclosed herein, creating a composition/detectable moiety conjugate.
  • the compositions disclosed herein may be labeled using labels and techniques known to one of ordinary skill in the art. Some of these labels are described in the "Handbook of Fluorescent Probes and Research Products", ninth edition, Richard P. Haugland (ed) Molecular Probes, Inc. Eugene, OR), which is incorporated herein in its entirety.
  • Detectable moieties suitable for such use include any composition detectable by spectroscopic, photochemical, biochemical, immunochemical, electrical, optical, magnetic or chemical means.
  • the detectable moieties contemplated for the present disclosure can include, but are not limited to, an immunofluorescent moiety (e.g., fluorescein, rhodamine, Texas red, and the like), a radioactive moiety (e.g., 3 H, 32 P, 125 1, 131 1, 35 S), an enzyme moiety (e.g., horseradish peroxidase, alkaline phosphatase), a colorimetric moiety (e.g., colloidal gold, biotin, colored glass or plastic, and the like).
  • the detectable moiety can be linked to the composition.
  • a linker can be included between the composition and the detectable moiety.
  • radiolabels may be detected using photographic film, gamma counters or scintillation counters.
  • Fluorescent markers may be detected using a photodetector to detect emitted illumination.
  • Enzymatic labels are typically detected by providing the enzyme with a substrate and detecting the reaction product produced by the action of the enzyme on the substrate, and colorimetric labels are detected by simply visualizing the colored label.
  • HDL mimetic peptides peptides or peptide analogs of the invention can be prepared using virtually any technique known to one of ordinary skill in the art for the preparation of peptides.
  • the peptides can be prepared using step-wise solution or solid phase peptide syntheses, or recombinant DNA techniques, or the equivalents thereof
  • HDL mimetic peptides of the invention containing amino acids having either the D- or L- configuration can be readily synthesized by automated solid phase procedures well known in the art. Suitable syntheses can be performed by utilizing "T-boc" or "F-moc” procedures. Techniques and procedures for solid phase synthesis are described in Solid Phase Peptide Synthesis: A Practical Approach, by E. Atherton and R. C. Sheppard, published by IRL, Oxford University Press, 1989. Alternatively, the peptides may be prepared by way of segment condensation, as described, for example, in Liu et ah, Tetrahedron Lett. 37:933-936, 1996; Baca et al, J. Am. Chem. Soc.
  • Bodanszky M. and Bodanszky, A., The Practice of Peptide Synthesis, Springer Verlag, New York, 1994; and by Jones, J., Amino Acid and Peptide Synthesis, 2nd ed., Oxford University Press, 2002.
  • the Bodanszky and Jones references detail the parameters and techniques for activating and coupling amino acids and amino acid derivatives. Moreover, the references teach how to select, use and remove various useful functional and protecting groups.
  • Peptides of the disclosure having either the D- or L-configuration can also be readily purchased from commercial suppliers of synthetic peptides. Such suppliers include, for example, Advanced ChemTech (Louisville, KY), Applied Biosystems (Foster City, CA), Anaspec (San Jose, CA), and Cell Essentials (Boston, MA).
  • the peptide or the relevant portion can also be synthesized using conventional recombinant genetic engineering techniques.
  • a polynucleotide sequence encoding the peptide is inserted into an appropriate expression vehicle, that is, a vector which contains the necessary elements for the transcription and translation of the inserted coding sequence, or in the case of an RNA viral vector, the necessary elements for replication and translation.
  • the expression vehicle is then transfected into a suitable target cell which will express the peptide.
  • the expressed peptide is then isolated by procedures well-established in the art.
  • the polynucleotide can be designed to encode multiple units of the peptide separated by enzymatic cleavage sites.
  • the resulting polypeptide can be cleaved ⁇ e.g., by treatment with the appropriate enzyme) in order to recover the peptide units.
  • This can increase the yield of peptides driven by a single promoter.
  • a polycistronic polynucleotide can be designed so that a single mRNA is transcribed which encodes multiple peptides, each coding region operatively linked to a cap-independent translation control sequence, for example, an internal ribosome entry site (IRES).
  • IRS internal ribosome entry site
  • the translation of each peptide encoded by the mRNA is directed internally in the transcript, for example, by the IRES.
  • the polycistronic construct directs the transcription of a single, large polycistronic mRNA which, in turn, directs the translation of multiple, individual peptides. This approach eliminates the production and enzymatic processing of polyproteins and can significantly increase yield of peptide driven by a single promoter.
  • a variety of host-expression vector systems may be utilized to express the peptides described herein.
  • microorganisms such as bacteria transformed with recombinant bacteriophage DNA or plasmid DNA expression vectors containing an appropriate coding sequence; yeast or filamentous fungi transformed with recombinant yeast or fungi expression vectors containing an appropriate coding sequence; insect cell systems infected with recombinant virus expression vectors (e.g., baculovirus) containing an appropriate coding sequence; plant cell systems infected with recombinant virus expression vectors (e.g., cauliflower mosaic virus (CaMV) or tobacco mosaic virus (TMV)) or transformed with recombinant plasmid expression vectors (e.g., Ti plasmid) containing an appropriate coding sequence; or animal cell systems.
  • the expression elements of the expression systems vary in their strength and specificities.
  • any of a number of suitable transcription and translation elements can be used in the expression vector.
  • inducible promoters such as pL of bacteriophage lambda, plac, ptrp, ptac (ptrp-lac hybrid promoter) and the like can be used.
  • promoters such as the baculovirus polyhedron promoter can be used.
  • promoters derived from the genome of plant cells e.g., heat shock promoters, the promoter for the small subunit of RUBISCO, the promoter for the chlorophyll a/b binding protein
  • plant viruses e.g., the 35S RNA promoter of CaMV, the coat protein promoter of TMV
  • promoters derived from the genome of mammalian cells e.g., metallothionein promoter
  • mammalian viruses e.g., the adenovirus late promoter, the vaccinia virus 7.5 K promoter
  • the HDL mimetic peptides or peptide analogs of the invention can be purified by many techniques well known in the art, such as reverse phase chromatography, high performance liquid chromatography, ion exchange chromatography, size exclusion chromatography, affinity chromatography, gel electrophoresis, and the like.
  • the actual conditions used to purify a particular peptide or peptide analog will depend, in part, on synthesis strategy and on factors such as net charge, hydrophobicity, hydrophilicity, and the like, and will be apparent to those of ordinary skill in the art.
  • any antibody which specifically binds the peptide or peptide analog may be used.
  • the peptides of the present invention may optionally be acetylated at the N-terminus.
  • the peptides of the present invention may optionally have a carboxy terminal amide.
  • the peptides of the present invention may have both an acetylated N-terminus and a carboxy terminal amide. Methods of acetylating the N-terminus or adding a carboxy terminal amide are well known to one of ordinary skill in the art.
  • compositions of the disclosure can be used, alone or in combination, together with a pharmaceutically acceptable carrier, to treat any disorder in animals, especially mammals (e.g., humans), for increasing HDL levels or promoting lipid efflux is beneficial.
  • disorders include, but are not limited to, dyslipidemic and vascular disorders.
  • the dyslipidemic and vascular disorders include hyperlipidemia (e.g., hypercholesterolemia), hyperlipoproteinemia, hypercholesterolemia, hypertriglyceridemia, HDL deficiency, apoAl deficiency, cardiovascular disease, cerebrovascular disease, coronary artery disease, atherosclerosis, myocardial infarction, stroke, thrombotic stroke, peripheral vascular disease, restenosis (e.g., atherosclerotic plaques), acute coronary syndrome, and reperfusion myocardial injury.
  • hyperlipidemia e.g., hypercholesterolemia
  • hyperlipoproteinemia e.g., hypercholesterolemia
  • hypertriglyceridemia e.g., HDL deficiency
  • apoAl deficiency e.g., apoAl deficiency
  • cardiovascular disease cerebrovascular disease
  • coronary artery disease atherosclerosis
  • myocardial infarction stroke
  • thrombotic stroke thrombotic stroke
  • compositions of the disclosure can also be used alone or in combination during the treatment of thrombotic stroke, infarcts secondary to occlusion of a vessel and during thrombolytic treatment of occluded coronary artery disease.
  • the compositions of the disclosure can be used to treat tissue following hypoxia, ischemia and infarction due to impairment of blood supply, and also following hemorrhage following rupture or trauma of a blood vessel.
  • tissue includes, without limitation, neural tissue in the central or peripheral nervous system, peripheral vascular tissue, and cardiac muscle.
  • the present compositions can be used alone or in combination therapy with other lipid- lowering compositions or drugs and/or other anti-inflammatory compositions or drugs used to treat the foregoing conditions.
  • Such therapies include, but are not limited to simultaneous or sequential administration of the drugs involved.
  • the pharmaceutical composition formulations can be administered with any one or more of the cholesterol lowering therapies currently in use, for example, bile-acid resins, niacin, statins, fat uptake inhibitors, and HDL-raising drugs.
  • the present compositions can be used in conjunction with statins or fibrates to treat hyperlipidemia, hypercholesterolemia and/or cardiovascular disease, such as atherosclerosis.
  • compositions of the disclosure can be used in combination with an anti-microbial agent and/or an anti-inflammatory agent, such as aspirin.
  • compositions of the disclosure can be used in combination with anti- hypertensive medicines known to one of ordinary skill in the art. It is to be understood that more than one additional therapy may be combined with administration of the compositions of the disclosure.
  • the present compositions can be used in conjunction with medicines used to treat patients with cerebrovascular and cardiovascular disease resulting in hypoxia, ischemia and infarction due to impairment of blood supply, and also following hemorrhage following rupture or trauma of a blood vessel.
  • medicines are commonly known to one of ordinary skill in the art and include without limitation, modulators of excitatory amino acids and modulators of platelet aggregation.
  • compositions comprising the compositions of the present invention in an acceptable carrier are administered to an animal or a human suffering from a dyslipidemic or vascular disorder, such as hyperlipidemia, hyperlipoproteinemia, hypercholesterolemia, hypertriglyceridemia, HDL deficiency, apoAl deficiency, coronary artery disease, atherosclerosis, stroke, ischemia, infarction, myocardial infarction, hemorrhage, peripheral vascular disease, restenosis, acute coronary syndrome, or reperfusion myocardial injury, in an amount sufficient to inhibit or treat the dyslipidemic or vascular disorder.
  • Amounts effective for this use will depend upon the severity of the disorder and the general state of the subject's health.
  • a therapeutically effective amount of the compound is that which provides either subjective relief of a symptom(s) or an objectively identifiable improvement as noted by the clinician or other qualified observer.
  • a pharmaceutical composition can be administered by any means known to one of skill in the art, such as by oral, intramuscular, subcutaneous, intracardiac, intraarterial or intravenous injection, but even nasal or anal administration is contemplated.
  • administration is by oral administration.
  • administration is by subcutaneous or intramuscular injection.
  • the composition can be provided as an implant, an oily injection, or as a particulate system.
  • the particulate system can be a vesicle, microparticle, a microcapsule, a microsphere, a nanoparticle, or similar particle (Banga, "Parenteral Controlled Delivery of Therapeutic Peptides and Proteins," in Therapeutic Peptides and Proteins, Technomic Publishing Co., Inc., Lancaster, PA, 1995).
  • the present compositions may also be applied to a medical device for delivery to a specific location.
  • a stent, a surgical tool, catheter, balloon, electrode, suture, or an artificial vessel or transplanted vessel may contain or be coated with the present composition.
  • vesicles, such as nanoparticles containing the HDL mimetic peptides are embedded into the stent and released when the stent is located within a desired vessel to promote lipid efflux and/or HDL release.
  • compositions of the present invention can be combined with a pharmaceutically acceptable carrier ⁇ e.g., a phospholipid or other type of lipid) or vehicle for administration to human or animal subjects.
  • a pharmaceutically acceptable carrier e.g., a phospholipid or other type of lipid
  • more than one composition can be combined to form a single preparation.
  • the compositions can be conveniently presented in unit dosage form and prepared using conventional pharmaceutical techniques. Such techniques include the step of bringing into association the active ingredient and the pharmaceutical carrier(s) or excipient(s). In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers.
  • Formulations suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents.
  • the formulations may be presented in unit- dose or multi-dose containers, for example, sealed ampules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of a sterile liquid carrier, for example, water for injections, immediately prior to use.
  • Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets commonly used by one of ordinary skill in the art.
  • unit dosage formulations are those containing a dose or unit, or an appropriate fraction thereof, of the administered ingredient. It should be understood that in addition to the ingredients particularly mentioned above, formulations encompassed herein may include other agents commonly used by one of ordinary skill in the art.
  • compositions provided herein may be administered through different routes, such as oral, including buccal and sublingual, rectal, parenteral, aerosol, nasal, intramuscular, intraperitoneal, intravascular, subcutaneous, intradermal, and topical. They may be administered in different forms, including but not limited to solutions, emulsions and suspensions, microspheres, vesicles, particles, microparticles, nanoparticles, and liposomes.
  • This maybe achieved by, for example, and not by way of limitation, local or regional infusion or perfusion during surgery, direct perfusion into the heart, particularly the left ventricle, direct perfusion into a vessel, such as the aorta, celiac trunk, common hepatic artery, right or left hepatic artery, portal vein or any of the veins leading to the portal vein, an atherosclerotic vessel, topical application (e.g., wound dressing, coated stent), injection, catheter, suppository, or implant (e.g., implants formed from porous, non- porous, or gelatinous materials, including membranes, such as silastic membranes or fibers), and the like.
  • a vessel such as the aorta, celiac trunk, common hepatic artery, right or left hepatic artery, portal vein or any of the veins leading to the portal vein, an atherosclerotic vessel
  • administration can be by direct injection at the site (or former site) of a tissue that is to be treated, such as the liver, heart or the peripheral vasculature.
  • the pharmaceutical compositions are delivered in a vesicle, in particular liposomes (see, e.g., Langer, Science 249:1527-1533, 1990; Treat et al, in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, N. Y., pp. 353-365, 1989) or nanoparticles.
  • Combinations of administration methods may also be employed such as oral, systemic and/or local infusion of a pharmaceutical composition of the present invention, before, after or during placement of a stent coated with a pharmaceutical composition of the present invention.
  • Stents containing the compositions of the present invention may be used at any location where treatment is desired, e.g. but not limited to a coronary artery or a carotid artery. Any vessel in the cardiovascular system that needs treatment may be treated.
  • the pharmaceutical compositions can be delivered in a controlled release system.
  • a pump can be used (see, e.g., Longer Science 249:1527-1533, 1990; Sefton Crit. Rev. Biomed. Eng. 14:201-240, 1987; Buchwald et al, Surgery 88:507-516, 1980; Saudek et al, N. Engl. J. Med. 321:574-579, 1989).
  • polymeric materials can be used (see, e.g., Ranger et al, Macromol ScL Rev. Macromol. Chem. 23:61-64, 1983; Levy et al, Science 228:190-192, 1985; During et al, Ann.
  • the amount of the pharmaceutical compositions that will be effective depends on the nature of the disorder or condition to be treated, as well as the stage of the disorder or condition. Effective amounts can be determined by standard clinical techniques. The precise dose to be employed in the formulation will also depend on the route of administration, and should be decided according to the judgment of the health care practitioner and each subject's circumstances. All dosage forms and therapeutically effective amounts of the compositions of the present invention may be employed. An example of such a dosage range is 0.1 to 200 mg/kg body weight in single or divided doses. Another example of a dosage range is 1.0 to 100 mg/kg body weight in single or divided doses.
  • the specific dose level and frequency of dosage for any particular subject may be varied and will depend upon a variety of factors, including the activity of the specific compound, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, and severity of the condition of the subject undergoing therapy.
  • compositions of the present disclosure can be administered at about the same dose throughout a treatment period, in an escalating dose regimen, or in a loading-dose regime (e.g., in which the loading dose is about two to five times the maintenance dose).
  • the dose is varied during the course of a treatment based on the condition of the subject being treated, the severity of the disease or condition, the apparent response to the therapy, and/or other factors as judged by one of ordinary skill in the art.
  • the volume of administration will vary depending on the route of administration.
  • oral administration may employ any orally acceptable dosage form commonly known to one of ordinary skill in the art (pill, lozenge, flash melt formulation, time release capsules, liquid, syrup, emulsion, etc.) and any therapeutically effective amount of the compositions of the present invention.
  • intramuscular injections may range from about 0.1 ml to about 1.0 ml. Those of ordinary skill in the art will know appropriate volumes for different routes of administration.
  • Example 1 Administration of the compositions of the present invention to increase lipid efflux and HDL in humans
  • An individual with low plasma levels of apoAl and HDL, relatively high levels of LDL, and high cholesterol is treated with the pharmaceutical compositions of the present invention through a daily oral administration. After a week, plasma levels of HDL are increased and total cholesterol is decreased.
  • Example 2 Intravenous administration of the compositions of the present invention to treat atherosclerosis in humans
  • IVUS intravascular ultrasound
  • Each individual is on stable hypolipidemic drug therapy and receives an acceptable dose of a pharmaceutical composition of the present invention iv weekly for a period of 5 to 8 weeks.
  • a repeat IVUS measurement is made at the end of the treatment period to assess the effect of the pharmaceutical composition infusion on coronary atherosclerosis in the target vessel.
  • Plaque is reduced in the atherosclerotic coronary artery following the treatment, demonstrating efficacy of the pharmaceutical compositions of the present invention to treat atherosclerosis.
  • Example 3 Oral administration of the compositions of the present invention to treat atherosclerosis in humans
  • IVUS intravascular ultrasound
  • Example 4 Administration of the pharmaceutical compositions of the present invention to prevent atherosclerosis in humans
  • Individuals with documented risk factors for atherosclerosis and having high plasma cholesterol levels have a ultrasound analysis of the coronary (IVUS), carotid (IMT), or popliteal arteries to establish a baseline measurement.
  • IVUS ultrasound analysis of the coronary
  • IMT carotid
  • popliteal arteries to establish a baseline measurement.
  • a portion of these individuals are daily administered individual pharmaceutical compositions of the present invention at a dose of 2 mg/kg to 50 mg/kg intravenously (iv) or intramuscularly (im) 1 to 3 times per week over a period of approximately one to six months.
  • each individual receives a suitable oral dose of such a pharmaceutical composition daily for approximately one to six months.
  • Example 5 Administration of the compositions of the present invention on stents and/or orally, to reduce inflammation and restenosis
  • One group receives a stent coated with a peptide of the present invention.
  • a second group receives an oral administration of the composition of the present invention at a dose of 2 mg/kg to 50 mg/kg, 2 to 3 times per week over a period of approximately 6 to 10 weeks.
  • a third group receives a stent coated with a peptide of the present invention and an oral administration of the composition of a peptide of the present invention at a dose of 2 mg/kg to 50 mg/kg, 2 to 3 times per week over a period of approximately 6 to 10 weeks.
  • Example 6 Administration of the pharmaceutical compositions of the present invention to prevent myocardial infarction or stroke in humans
  • a first group receives a stent coated with an encapsulated peptide of the present invention.
  • a second group receives an orally-administered encapsulated peptide of the present invention at a dose of about 2 mg/kg to 50 mg/kg, two to three times per week over a period of about six months to about thirty-six months.
  • a third group receives the coated stent plus the orally administered encapsulated peptide.
  • a fourth group receives an uncoated stent.
  • a fifth group receives an orally administered control composition (e.g., a placebo).

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Abstract

The present invention relates to novel compositions, uses of these compositions in the preparation of a medicament, and methods of administering these compositions to treat dyslipidemic and vascular disorders in humans and animals. The compositions comprise vesicles encapsulating high density lipoprotein (HDL) mimetic peptides that stimulate lipid efflux and/or increase HDL levels in an animal or a human. Increased HDL levels provide numerous health benefits, particularly cardiovascular health benefits.

Description

ENCAPSULATED HDL MIMETIC PEPTIDES
FIELD OF THE INVENTION
The present invention relates to novel compositions and methods of administering these compositions to treat dyslipidemic and vascular disorders in humans and animals. The compositions are vesicles encapsulating high density lipoprotein (HDL) mimetic peptides that stimulate lipid efflux and/or increase HDL levels in an animal or a human. Increased HDL levels provide numerous health benefits, particularly cardiovascular health benefits.
BACKGROUND OF THE INVENTION
Cardiovascular disease is a major cause of mortality and morbidity, especially in countries of the Western world. Dyslipidemic and vascular disorders affect numerous individuals throughout the world. Diseases such as hyperlipidemia, hyperlipoproteinemia, hypercholesterolemia, hypertriglyceridemia, HDL deficiency, apoAl deficiency, coronary artery disease, atherosclerosis, myocardial infarction, stroke and inflammation secondary to stroke, ischemia, ischemic stroke, thrombotic stroke, peripheral vascular disease, restenosis, thrombosis, acute coronary syndrome, and reperfusion myocardial injury cause suffering and death. In some individuals, plasma levels of apoAl and HDL are unacceptably low, while other molecules such as low density lipoprotein (LDL) remain high. Many of these individuals are at risk for significant cardiovascular disease and heart attacks.
What is needed are new compositions that increase lipid efflux and HDL levels in a recipient of the composition. What is also needed are new compositions that are effective in treating these conditions or in slowing the progression of disease.
SUMMARY OF THE INVENTION
The present invention provides novel compositions and methods of administering these compositions to animals or humans in order to increase lipid efflux and increase HDL levels in the animal or human. In preferred embodiments, these compositions may be delivered orally or may be coated on stents for delivery to blood vessels. The novel compositions comprise a vesicle encapsulating an HDL mimetic peptide. In several embodiments, these novel compositions are combined with a pharmaceutically acceptable carrier to make a pharmaceutical composition which may be administered to an individual to promote lipid efflux. The compositions of the invention can be used to manufacture medicaments for increasing lipid efflux, increasing HDL levels, and/or treating a dyslipidemic or vascular disorder in an individual receiving the medicament. Medicaments containing compositions of the invention can be manufactured for oral administration, for coating a vascular stent, for intravenous or intramuscular administration, or for administration by any other means known in the art and/or set forth herein. Administration of the compositions of the invention to individuals in need thereof can be through any appropriate means, including but not limited to oral administration or as a coating on a stent.
An individual who may be treated by administering the compositions of the present invention includes any individual who may benefit from an increase in lipid efflux or an increase in plasma HDL levels. Such individuals may have a dyslipidemic or vascular disease. Other individuals who may be treated have no clinical manifestation of disease but may be prone to such disease through genetic and/or environmental factors. Treatment of these individuals helps to delay or prevent the onset of dyslipidemic or vascular disease.
Administration of these pharmaceutical compositions may be through any means described herein and includes, but is not limited to, oral administration or as a coating on a stent.
Also described herein is a method of treating dyslipidemic and vascular disorders in an animal or a human, including administering to the animal or the human a therapeutically effective amount of the compositions of the present invention. Dyslipidemic and vascular disorders amenable to treatment with the compositions disclosed herein include, but are not limited to, hyperlipidemia, hyperlipoproteinemia, hypercholesterolemia, hypertriglyceridemia, HDL deficiency, apoAl deficiency, coronary artery disease, atherosclerosis, myocardial infarction, stroke and inflammation secondary to stroke, ischemia, ischemic stroke, thrombotic stroke, peripheral vascular disease, restenosis, thrombosis, acute coronary syndrome, and reperfusion myocardial injury. Accordingly, it is an object of the present invention to provide novel compositions comprising a vesicle encapsulating an HDL mimetic peptide.
It is another object of the present invention to provide pharmaceutical compositions comprising a vesicle encapsulating an HDL mimetic peptide, in combination with a pharmaceutically acceptable carrier. Another object of the present invention is to provide a stent coated with a novel composition comprising a vesicle encapsulating an HDL mimetic peptide.
It is another object of the present invention to provide a stent coated with a pharmaceutical composition comprising a vesicle encapsulating an HDL mimetic peptide in combination with a pharmaceutically acceptable carrier. It is another object of the present invention to provide compositions comprising encapsulated peptides for use in the manufacture of medicaments for increasing lipid efflux, increasing HDL levels, and/or treating a dyslipidemic or vascular disorder in an individual receiving such a medicament. Another object of the present invention is to provide methods of increasing lipid efflux and increasing HDL levels in an animal or a human comprising administration of the pharmaceutical compositions of the present invention to the animal or the human.
Another object of the present invention is to provide methods of increasing HDL levels in an animal or a human comprising administration of the pharmaceutical compositions of the present invention to the animal or the human.
Yet another object of the present invention is to provide methods of treating dyslipidemic or vascular disease or slowing the progression of dyslipidemic or vascular disease comprising administration of the pharmaceutical compositions of the present invention to an animal or a human in need thereof. It is yet another object of the present invention to provide new methods for the treatment of dyslipidemic or vascular disease in an animal or a human by administering the pharmaceutical compositions of the present invention.
It is yet another object of the present invention to provide new methods for the treatment of atherosclerosis, cardiovascular disease and cerebrovascular disease in an animal or a human by administering the pharmaceutical compositions of the present invention.
These and other objects, features and advantages of the present invention will become apparent after a review of the following detailed description of the disclosed embodiments and claims.
DETAILED DESCRIPTION OF THE INVENTION
The present invention provides novel compositions comprising a vesicle encapsulating an
HDL mimetic peptide.
The compositions of the present invention may be combined with a pharmaceutically acceptable carrier to form a pharmaceutical composition and administered to an animal or a human. These compositions cause increased lipid efflux and increase HDL levels. By increasing HDL levels, this therapeutic method treats conditions in which raising HDL levels is indicated.
Compositions of the invention can be administered by any appropriate method as described herein, including but not limited to, oral administration or administration as a coating on a vascular stent inserted into an individual's blood vessel. An individual who may be treated by administering the compositions of the present invention includes any individual who may benefit from an increase in lipid efflux or an increase in plasma HDL levels. Such individuals may have a dyslipidemic or vascular disease. Other individuals who may be treated have no clinical manifestation of disease but may be prone to such disease through genetic and/or environmental factors. Treatment of these individuals helps to delay or prevent the onset of dyslipidemic or vascular disease.
Administration of these pharmaceutical compositions may be through any means described herein and includes, but is not limited to, oral administration and administration as a coating on a stent. Also described herein is a method of treating dyslipidemic and vascular disorders in an animal or a human, including administering to the animal or the human a therapeutically effective amount of the compositions of the present invention. Dyslipidemic and vascular disorders amenable to treatment with the compositions disclosed herein include, but are not limited to, hyperlipidemia, hyperlipoproteinemia, hypercholesterolemia, hypertriglyceridemia, HDL deficiency, apoAl deficiency, coronary artery disease, atherosclerosis, myocardial infarction, stroke and inflammation secondary to stroke, ischemia, ischemic stroke, thrombotic stroke, peripheral vascular disease, restenosis, thrombosis, acute coronary syndrome, and reperfusion myocardial injury.
/. Abbreviations
ABCAl : ATP-binding cassette transporter Al apoAl : apolipoprotein Al
DMPC: dimyristoyl phosphatidyl choline
HDL: high-density lipoprotein HPLC: high-pressure liquid chromatography
LDL: low-density lipoprotein
RBC: red blood cell
//. Terms Unless otherwise noted, technical terms are used according to conventional usage.
Definitions of common terms in molecular biology may be found in Benjamin Lewin, Genes VII, published by Oxford University Press, 2000 (ISBN 019879276X); Kendrew et al. (eds.), The Encyclopedia of Molecular Biology, published by Blackwell Publishers, 1994 (ISBN 0632021829); and Robert A. Meyers (ed.), Molecular Biology and Biotechnology: a Comprehensive Desk Reference, published by Wiley, John & Sons, Inc., 1995 (ISBN 0471186341); and other similar references.
As used herein, the singular terms "a," "an," and "the" include plural referents unless context clearly indicates otherwise. Similarly, the word "or" is intended to include "and" unless the context clearly indicates otherwise. Also, as used herein, the term "comprises" means "includes." Hence "comprising A or B" means including A, B, or A and B.
In order to facilitate review of the various embodiments of this disclosure, the following explanations of specific terms are provided:
Analog, derivative or mimetic: An analog is a molecule that differs in chemical structure from a parent compound, for example a homolog (differing by an increment in the chemical structure, such as a difference in the length of an alkyl chain), a molecular fragment, a structure that differs by one or more functional groups, a change in ionization. Structural analogs are often found using quantitative structure activity relationships (QSAR), with techniques such as those disclosed in Remington (The Science and Practice of Pharmacology, 19th Edition (1995), chapter 28). A derivative is a biologically active molecule derived from the base structure. A mimetic is a molecule that mimics the activity of another molecule, such as a biologically active molecule.
Biologically active molecules can include chemical structures that mimic the biological activities of a compound.
Animal: Living multi-cellular vertebrate organisms, a category that includes, for example, mammals and birds. The term mammal includes both human and non-human mammals. Similarly, the term "subject" includes both human and veterinary subjects, for example, humans, non-human primates, dogs, cats, horses, and cows.
Antibody: A protein (or protein complex) that includes one or more polypeptides substantially encoded by immunoglobulin genes or fragments of immunoglobulin genes. The recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon, and mu constant region genes, as well as the myriad immunoglobulin variable region genes. Light chains are classified as either kappa or lambda. Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which in turn define the immunoglobulin classes, IgG, IgM, IgA, IgD and IgE, respectively. Domain: A domain of a protein is a part of a protein that shares common structural, physiochemical and functional features; for example hydrophobic, polar, globular, helical domains or properties, for example a DNA binding domain, an ATP binding domain, and the like. Dyslipidemic disorder: A disorder associated with any altered amount of any or all of the lipids or lipoproteins in the blood. Dyslipidemic disorders include, for example, hyperlipidemia, hyperlipoproteinemia, hypercholesterolemia, hypertriglyceridemia, HDL deficiency, apoAl deficiency, and cardiovascular disease (e.g., coronary artery disease, atherosclerosis and restenosis).
Efflux: The process of flowing out. As applied to the results described herein, lipid efflux refers to a process whereby lipid, such as cholesterol and phospholipid, is complexed with an acceptor, such as an apolipoprotein or apolipoprotein peptide mimetic, and removed from vesicles or cells. "ABCAl -dependent lipid efflux" (or lipid efflux by an "ABCAl-dependent pathway") refers to a process whereby apolipoproteins or synthetic peptide mimetics of apolipoproteins bind to a cell and efflux lipid from the cell by a process that is facilitated by the ABCAl transporter.
Inhibiting or treating a disease: Inhibiting the full development of a disease, disorder or condition, for example, in a subject who is at risk for a disease such as atherosclerosis and cardiovascular disease. "Treatment" refers to a therapeutic intervention that ameliorates a sign or symptom of a disease or pathological condition after it has begun to develop. As used herein, the term "ameliorating," with reference to a disease, pathological condition or symptom, refers to any observable beneficial effect of the treatment. The beneficial effect can be evidenced, for example, by a delayed onset of clinical symptoms of the disease in a susceptible subject, a reduction in severity of some or all clinical symptoms of the disease, a slower progression of the disease, a reduction in the number of relapses of the disease, an improvement in the overall health or well-being of the subject, or by other parameters well known in the art that are specific to the particular disease.
Isolated/purified: An "isolated" or "purified" biological component (such as a nucleic acid, peptide or protein) has been substantially separated, produced apart from, or purified away from other biological components in the cell of the organism in which the component naturally occurs, that is, other chromosomal and extrachromosomal DNA and RNA, and proteins. Nucleic acids, peptides and proteins that have been "isolated" thus include nucleic acids and proteins purified by standard purification methods. The term also embraces nucleic acids, peptides and proteins prepared by recombinant expression in a host cell as well as chemically synthesized nucleic acids or proteins. The term "isolated" or "purified" does not require absolute purity; rather, it is intended as a relative term. Thus, for example, an isolated biological component is one in which the biological component is more enriched than the biological component is in its natural environment within a cell. Preferably, a preparation is purified such that the biological component represents at least 50%, such as at least 70%, at least 90%, at least 95%, or greater of the total biological component content of the preparation.
Label: A detectable compound or composition that is conjugated directly or indirectly to another molecule to facilitate detection of that molecule. Specific, non-limiting examples of labels include fluorescent tags, colorimetric labels, dyes, beads, enzymatic linkages, radiodense materials, and radioactive isotopes.
Ligand: A molecule that can bind to another molecule.
Linker: A molecule that joins two other molecules, either covalently, or through ionic, van der Waals or hydrogen bonds. Lipid: A class of water-insoluble, or partially water insoluble, oily or greasy organic substances, that are extractable from cells and tissues by nonpolar solvents, such as chloroform or ether. Types of lipids include triglycerides (e.g., natural fats and oils composed of glycerin and fatty acid chains), glycolipids, phospholipids (e.g., phosphatidylethanolamine, phosphatidylcholine, phosphatidylserine, and phosphatidylinositol), sphingolipids (e.g., sphingomyelin, cerebrosides and gangliosides), and sterols (e.g., cholesterol).
Non-cytotoxic: A non-cytotoxic compound is one that does not substantially affect the viability or growth characteristics of a cell at a dosage normally used to treat the cell or a subject. Furthermore, the percentage of cells releasing intracellular contents, such as LDH or hemoglobin, is low (e.g., about 10% or less) in cells treated with a non-cytotoxic compound. Lipid efflux from a cell that occurs by a non-cytotoxic compound results in the removal of lipid from a cell by a process that maintains the overall integrity of the cell membrane and does not lead to significant cell toxicity.
Non-polar: A non-polar compound is one that does not have concentrations of positive or negative electric charge. Non-polar compounds, such as, for example, oil, are not well soluble in water.
Peptide: A polymer in which the monomers are amino acid residues which are joined together through amide bonds. When the amino acids are alpha-amino acids, either the L-optical isomer or the D-optical isomer can be used. The amino acid sequences disclosed herein are shown using standard single letter abbreviations as known to one of ordinary skill in the art, and in some cases as three letter codes for amino acids, as defined in 37 C.F.R. 1.822 and as commonly known to one of ordinary skill in the art. When the single letter designation for an amino acid, for example S for serine is shown in lower case, s, the serine is a D amino acid. The terms "peptide" or "polypeptide" as used herein are intended to encompass any amino acid sequence and include modified sequences such as glycoproteins. The term "peptide" is specifically intended to cover naturally occurring peptides, as well as those which are recombinantly or synthetically produced. The term "residue" or "amino acid residue" includes reference to an amino acid that is incorporated into a peptide, polypeptide, or protein. As known to one of skill in the art, the peptides presented herein are read from the N to the C terminus i.e., from left to right. Accordingly, the N terminal amino acid in Leu GIu Ser is Leu and the C- terminal amino acid is Ser.
Substitutions: Peptides of the present invention include peptides with substitutions for amino acids in the peptide sequence. Such substitutions may be conservative substitutions, isosteric substitutions, substitutions between isosteric groups, and non-conservative substitutions as defined herein.
Peptides of the present invention include conservatively substituted peptides, wherein these conservative substitutions occur at 1%, 3%, 5%, 7%, 10%, 15%, 20%, 25%, 30%, 40%, or 50% of the amino acid residues. Peptides of the present invention include peptides that are homologous at 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99% of the entire sequence of the peptide.
Pharmaceutically acceptable carriers: The pharmaceutically acceptable carriers (vehicles) useful in this disclosure are conventional. Remington 's Pharmaceutical Sciences, by E. W. Martin, Mack Publishing Co., Easton, PA, 15th Edition (1975), describes compositions and formulations suitable for pharmaceutical delivery of one or more therapeutic compounds or molecules, and additional pharmaceutical agents.
In general, the nature of the carrier will depend on the particular mode of administration being employed. For instance, parenteral formulations usually comprise injectable fluids that include pharmaceutically and physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or the like as a vehicle. For solid compositions (e.g., powder, pill, tablet, or capsule forms), conventional non-toxic solid carriers can include, for example, pharmaceutical grades of mannitol, lactose, starch, or magnesium stearate. In addition to biologically-neutral carriers, pharmaceutical compositions to be administered can contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, preservatives, and pH buffering agents and the like, for example sodium acetate or sorbitan monolaurate.
Phospholipid: A phospholipid consists of a water-soluble polar head, linked to two water- insoluble non-polar tails (by a negatively charged phosphate group). Both tails consist of a fatty acid, each about 14 to about 24 carbon groups long. When placed in an aqueous environment, phospholipids form a bilayer or micelle, where the hydrophobic tails line up against each other. This forms a membrane with hydrophilic heads on both sides. A phospholipid is a lipid that is a primary component of animal cell membranes.
Polar: A polar molecule is one in which the centers of positive and negative charge distribution do not converge. Polar molecules are characterized by a dipole moment, which measures their polarity, and are soluble in other polar compounds and virtually insoluble in nonpolar compounds.
Recombinant nucleic acid: A sequence that is not naturally occurring or has a sequence that is made by an artificial combination of two otherwise separated segments of sequence. This artificial combination is often accomplished by chemical synthesis or, more commonly, by the artificial manipulation of isolated segments of nucleic acids, for example, by genetic engineering techniques such as those described in Sambrook et al. (ed.), Molecular Cloning: A Laboratory Manual, 2nd ed., vol. 1-3, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989. The term recombinant includes nucleic acids that have been altered solely by addition, substitution, or deletion of a portion of the nucleic acid. Therapeutically effective amount: A quantity of a specified agent sufficient to achieve a desired effect in a subject being treated with that agent. For example, this can be the amount of a composition useful in preventing, ameliorating, and/or treating a dyslipidemic disorder {e.g., atherosclerosis) in a subject. Ideally, a therapeutically effective amount of an agent is an amount sufficient to prevent, ameliorate, and/or treat a dyslipidemic disorder (e.g., atherosclerosis) in a subject without causing a substantial cytotoxic effect (e.g., membrane microsolubilization) in the subject. The effective amount of an agent useful for preventing, ameliorating, and/or treating a dyslipidemic disorder (e.g., atherosclerosis) in a subject will be dependent on the subject being treated, the severity of the disorder, and the manner of administration of the therapeutic composition. Transformed: A "transformed" cell is a cell into which has been introduced a nucleic acid molecule by molecular biology techniques. The term encompasses all techniques by which a nucleic acid molecule might be introduced into such a cell, including transfection with viral vectors, transformation with plasmid vectors, and introduction of naked DNA by electroporation, lipofection, and particle gun acceleration.
///. Overview of Several Embodiments
The compositions of the present invention comprise a vesicle encapsulating an HDL mimetic peptide. These compositions may be combined with a pharmaceutically acceptable carrier to make a pharmaceutical composition for administration to an animal or a human. A. Vesicles for Use in the Compositions of the Present Invention
The present compositions comprise an HDL mimetic peptide encapsulated within a vesicle. The term "vesicle" includes vesicles of various sizes, including but not limited to nanoparticles. Thus, as used herein, the term "vesicle" also refers to nanoparticles. The vesicles of the present invention include, but are not limited to, lipid-containing vesicles (e.g., liposomes). For example, the vesicle can be a lipid vesicle as described in US Patent No. 5,985,852. Methods for preparing lipid vesicles containing peptides are well-known in the art, e.g., sonication methods, phase inversion methods, and oil/water emulsion methods. For example, the lipid vesicles containing peptides can be prepared as described in US Patent No. 5,916,588. In a brief example of a method for preparing lipid vesicles containing peptides, a stock solution of a peptide to be encapsulated is prepared at a concentration of 3 mg/ml in 50 mM NaPO4 buffer. A stock solution of a lipid such as dipalmityl phosphatidylcholine (DPPC) is prepared at a concentration of 20 mg/ml in chloroform:methanol (3:1). The peptide is combined with the lipid at a molar recombination ratio range of peptide to lipid from 1 :5 up to 1 :300. Appropriate aliquots of peptide and lipid are mixed and the sample is incubated at 50° C for about 30 minutes to about 3 hrs. to obtain clarity of the solution. Up to three cycles of heating and cooling may be required to obtain clarity of the sample. Following recombination, the sample is filtered over a polyethersulfone (PES) filter to remove unincorporated lipid. Lipids for incorporation into vesicles can be any suitable lipid or mixture of lipids, including, but not limited to, the electrically-neutral and electrically-charged phospholipids described hereinbelow.
Nanoparticles include, but are not limited to, those described in US Patent Application Serial No. 10/362,848 (US 2004/0063606), which describes biodegradable polymers, such as polyester amide polymers, polyester urethane polymers, and polyester urea polymers, that contain at least one amino acid and a non-amino acid moiety per repeat unit. Such vesicles and nanoparticles are well-known in the art and the present peptides are encapsulated into such nanoparticles according to well-known methods, e.g., as described in the foregoing publications. Hydrogel-type nanoparticles, as described, e.g., in US 2006/0177416 (see, e.g., page 1, paragraph 0006), can also be used in the compositions of the invention, for example, for compositions intended for oral administration. Other examples of nanoparticles suitable for oral administration include but are not limited to the nanoparticles described in Damge C. et al., Expert Opinion on Drug Delivery 5:45-68 (2008); Galindo-Rodriguez S. A., Critical Reviews in Therapeutic Drug Carrier Systems, 22:419-464 (2005); Dong Y. and Feng S.-S., Biomaterials 26:6068-6076 (2005); and Pamajula S. et al., J. Pharm. Pharmacol. 56:1119-1125 (2004). Such nanoparticles suitable for oral administration are well-known in the art and the present peptides are encapsulated into such nanoparticles according to well-known methods, e.g., as described in the foregoing publications.
An example of nanoparticles encapsulating an HDL mimetic peptide of the invention that are suitable for coating onto a vascular stent includes but is not limited to the poly-D,L-lactide- co-glycolic acid (PLGA) nanoparticles described in Nam SE, et al., Bull. Korean Chem. Soc. 28:397-402 (2007). Such PLGA nanoparticles are well-known in the art and the present peptides are encapsulated into such nanoparticles according to well-known methods, e.g., as described in Nam SE, et al, supra.
Methods for coating peptides onto stents are well-known in the art. For example, US Patent No. 7,037,552 (Coating Dispensing System and Method Using a Solenoid Head for Coating Medical Devices) provides methods for coating stents with liposomes or polymers containing a therapeutic agent (including a biological molecule, such as a protein). In another example, US Patent Publication No. 2004/0170685 (Bioactive Stents and Methods for Use Thereof) provides methods for coating stents with biodegradable polymers to which polypeptides are attached (see, e.g., paragraphs 0032-0034 and 0040). US Patent Publication No. 2004/0170685 also describes methods for producing biodegradable stents to which peptides are bound (see, e.g., paragraphs 0035-0040).
Other nanoparticles for use in the methods of the invention include those described in U.S. Patent Application Serial Nos. 11/344,689 (US 2006/0177416), 11/345,021 (US 2006/0188469), 11/345,815 (US 2006/0188486), and related applications. In one embodiment, the nanoparticles are chosen from those recited in the claims of US Patent Application Serial No. 10/362,848 (US 2004/0063606). Some appropriate nanoparticles for use in the compositions of the invention are disclosed in Reddy et al., Clin. Cancer Res. (2006) 12(22), 6677-6686. Peptides for use in the compositions of the invention are encapsulated into vesicles or nanoparticles using methods that are well-known in the art, e.g. but not limited to any of the methods set forth in the foregoing publications. The encapsulated peptides of the invention can be used for the manufacture of medicaments for use in increasing lipid efflux, increasing HDL levels, and/or treating a dyslipidemic or vascular disorder in an individual receiving the medicament. Medicaments containing peptides of the invention can be manufactured for oral administration, for coating a vascular stent, for intravenous or intramuscular administration, or for administration by any other means known in the art and/or set forth herein. Administration of the encapsulated peptides to individuals in need thereof can be through any appropriate means, including but not limited to oral administration or as a coating on a stent.
B. HDL Mimetic Peptides for Use in the Compositions of the Invention
The HDL mimetic peptides for use in the compositions present invention include the peptides set forth hereinbelow and as incorporated by reference. These applications incorporated by reference in their entirety include USSN 60/898,132, filed January 29, 2007, 11/764,619 filed June 18, 2007, 11/861,758 filed September 26, 2007, 61/023,963 filed January 28, 2008, 12/021,486 filed January 29, 2008, and 12/021,629 filed January 29, 2008. HDL mimetic peptides mimic naturally occuring ApoAI by binding lipid and by facilitating lipid efflux from cells. Peptides in the present compositions can be unmodified or modified as described herein and/or as will be appreciated by one of ordinary skill in the art.
The amino- and/or carboxy-termini of the peptides of the invention can be chemically modified by conjugation with various functional groups. One of skill in the art will appreciate that synthetic peptides carry free amino- and carboxy-termini, which carry, respectively, a positive and a negative charge. Particularly for (but not limited to) peptides designed to mimic an internal sequence of a protein, amino-terminal acetylation and/or carboxy-terminal amidation neutralizes, respectively, the amino-terminal and/or carboxy-terminal charge, such that the synthetic peptide more closely mimics the structure and function of the natural peptide. Such amino- and carboxy- terminal modification also renders peptides more resistant to enzymatic degradation, such as that resulting from exopeptidases. Neutralization of the terminal charge of synthetic peptide mimetics of apolipoproteins has been shown to increase their lipid affinity (Yancey et al., Biochem. 34:7955- 7965, 1995; Venkatachalapathi et al., Protein: Structure, Function and Genetics 15:349-359, 1993). For example, acetylation of the amino-terminus of amphipathic peptides increases the lipid affinity of the peptide (Mishra et al., J. Biol. Chem. 269:7185-7191, 1994). Other possible end modifications are described, for example, in Brouillette et al., Biochem. Biophys. Acta 1256:103- 129, 1995: Mishra et al., J. Biol. Chem. 269:7185-7191, 1994; and Mishra et al., J. Biol. Chem. 270:1602-1611, 1995. Non-limiting examples of the HDL mimetic peptides of the present invention modified by amino-terminal acetylation and/or carboxy-terminal amidation are set forth hereinbelow.
One of ordinary skill in the art will also understand that any of the peptides employed in compositions of the present invention may contain a D amino acid at the N-terminus, at the C- terminus, or at both the N- and C-termini. The presence of these D amino acids can help protect against peptide degradation, hi some embodiments, all the amino acids of any peptide of the present invention may be D amino acids. This embodiment is useful for protection against degradation following oral administration of a pharmaceutical composition comprising the peptides of the present invention. Some examples of peptides of the present invention containing one or more D amino acids include but are not limited to those shown hereinbelow. D amino acids are indicated in all-upper-case letters, e.g., "SER" or "PRO.
One of ordinary skill in the art will understand that the peptides for use in the present compositions can be modified by any other chemical modification that protects the present peptides from exopeptidase or endopeptidase digestion. In addition, the present peptides can be modified by a detectable moiety, as described herein and/or as will be appreciated by the skilled artisan.
One of ordinary skill in the art will further understand that any of the peptides used in compositions of the present invention can contain one or more amino acid substitutions, e.g., an isosteric amino acid substitution, a conservative amino acid substitution, and/or a non-conservative amino acid substitution as described hereinbelow.
i- Unmodified Peptides
SEQ ID NO: 1 Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn; SEQ ID NO: 2 Ser Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn;
SEQ ID NO: 3 Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys; SEQ ID NO: 4 Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp; SEQ ID NO: 5 Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn;
SEQ ID NO: 6 Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp;
SEQ ED NO: 7 Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn; SEQ ID NO: 8 Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp; SEQ ID NO: 9 Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn;
SEQ ID NO: 10 Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp; SEQ ID NO: 11 Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu Lys;
SEQ ID NO: 12 Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu Lys; SEQ ID NO: 13 Leu GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys Pro Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu Lys; SEQ ID NO: 14 Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu Lys; SEQ ID NO: 15 Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu Lys; SEQ ID NO: 16 Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys;
SEQ ID NO: 17 Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys; SEQ ID NO: 18 Asp Tip Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe Pro Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala Ala; SEQ ID NO: 19 Ala Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala Phe; SEQ ID NO: 20 Pro VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys GIn Lys Leu Lys.
ii. Peptides Containing an Amino-Terminal Acetylation
SEQ ID NO: 21 Ac-Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn; SEQ ID NO: 22 Ac-Ser Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn;
SEQ ID NO: 23 Ac-Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys; SEQ ID NO: 24 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Tip Trp Trp Trp;
SEQ ID NO: 25 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn;
SEQ ID NO: 26 Ac-Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp; SEQ ID NO: 27 Ac-Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn;
SEQ ID NO: 28 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp;
SEQ ID NO: 29 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn; SEQ ID NO: 30 Ac-Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Tip Arg Trp Trp Tip Tip;
SEQ ED NO: 31 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu Lys; SEQ ID NO: 32 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu Lys; SEQ ID NO: 33 Ac-Leu GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys Pro Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu Lys; SEQ ID NO: 34 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu Lys; SEQ ID NO: 35 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu Lys; SEQ ID NO: 36 Ac-Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys; SEQ ID NO: 37 Ac-Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys; SEQ ID NO: 38 Ac-Asp Trp Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe Pro Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala Ala; SEQ ID NO: 39 Ac-Ala Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala Phe; SEQ ID NO: 40 Ac-Pro VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys GIn Lys Leu Lys.
iii. Peptides Containing a Carboxy-Terminal Amidation SEQ ID NO: 41 Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn-NH2;
SEQ ID NO: 42 Ser Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn-NH2;
SEQ ID NO: 43 Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LyS-NH2; SEQ ID NO: 44 Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Tip Tip Trp Trp-NH2;
SEQ ID NO: 45 Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn-NH2;
SEQ ID NO: 46 Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Tip Arg Trp Trp Trp Trp-NH2; SEQ ID NO: 47 Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GkI-NH2;
SEQ ID NO: 48 Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp-NH2;
SEQ ID NO: 49 Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn-NH2; SEQ ID NO: 50 Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp-NH2;
SEQ ID NO: 51 Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LyS-NH2; SEQ ID NO: 52 Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LyS-NH2;
SEQ ID NO: 53 Leu GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys Pro Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu LyS-NH2; SEQ ID NO: 54 Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LyS-NH2; SEQ ID NO: 55 Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LyS-NH2; SEQ ID NO: 56 Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LyS-NH2; SEQ ID NO: 57 Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LyS-NH2; SEQ ID NO: 58 Asp Trp Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe Pro Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala AIa-NH2; SEQ ID NO: 59 Ala Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala Phe-NH2; SEQ ID NO: 60 Pro VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys GIn Lys Leu LyS-NH2.
iv. Peptides Containing an Amino-Terminal Acetylation and a Carboxy-Terminal Amidation SEQ ID NO: 61 Ac-Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn-NH2;
SEQ ID NO: 62 Ac-Ser Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn-NH2;
SEQ ID NO: 63 Ac-Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LyS-NH2; SEQ ID NO: 64 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp-NH2;
SEQ ID NO: 65 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn-NH2; SEQ ID NO: 66 Ac-Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp-NH2;
SEQ ID NO: 67 Ac-Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn-NH2;
SEQ ID NO: 68 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp-NH2; SEQ ID NO: 69 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala
Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe
Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn-NH2;
SEQ ID NO: 70 Ac-Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr
Lys Lys Ala Leu Ser Pro Leu Tip Arg Tip Tip Tip TIp-NH2;
SEQ ID NO: 71 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys
Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu Lys-
NH2; SEQ ID NO: 72 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys
Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LyS-NH2;
SEQ ID NO: 73 Ac-Leu GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys Pro
Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu LyS-NH2;
SEQ ID NO: 74 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu Lys-
NH2;
SEQ ID NO: 75 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys
Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LyS-NH2;
SEQ ID NO: 76 Ac-Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys-
NH2;
SEQ ID NO: 77 Ac-Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Pro
Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LyS-NH2;
SEQ ID NO: 78 Ac-Asp Tip Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe Pro Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala AIa-NH2;
SEQ ID NO: 79 Ac-Ala Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala Phe-
NH2;
SEQ ID NO: 80 Ac-Pro VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys
GIn Lys Leu LyS-NH2.
v. Peptides Containing an Amino-Terminal D Amino Acid
SEQ ID NO: 81 SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg
Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr
Lys Lys Leu Asn Thr GIn; SEQ ID NO: 82 SER Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys
GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr
Lys Lys Leu Asn Thr GIn;
SEQ ID NO: 83 SER Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys;
SEQ ID NO: 84 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg
Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys
VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp
Trp; SEQ ID NO: 85 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg
Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys
VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn;
SEQ ID NO: 86 SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg
Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp;
SEQ ID NO: 87 SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr
Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr
Lys Lys Leu Asn Thr GIn;
SEQ ID NO: 88 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys
VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp
Trp;
SEQ ID NO: 89 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys
VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn;
SEQ ID NO: 90 SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr
Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr
Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp;
SEQ ID NO: 91 LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu Lys;
SEQ ID NO: 92 LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro
Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu Lys;
SEQ ID NO: 93 LEU GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys Pro
Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu Lys; SEQ ID NO: 94 LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu Lys; SEQ ID NO: 95 LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu Lys; SEQ ID NO: 96 LEU GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys; SEQ ID NO: 97 LEU GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys; SEQ ID NO: 98 Ac-SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn;
SEQ ID NO: 99 Ac-SER Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn; SEQ ID NO: 100 Ac-SER Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys; SEQ ID NO: 101 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp;
SEQ ID NO: 102 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn; SEQ ID NO: 103 Ac-SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp;
SEQ ID NO: 104 Ac-SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn; SEQ ID NO: 105 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp; SEQ ID NO: 106 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala
Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe
Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn;
SEQ ID NO: 107 Ac-SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr
Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Tip Trp Trp Trp;
SEQ ID NO: 108 Ac-LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys
Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu Lys;
SEQ ID NO: 109 Ac-LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu Lys;
SEQ ID NO: 110 Ac-LEU GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys
Pro Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu Lys;
SEQ ID NO: 111 Ac-LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys
Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu Lys; SEQ ID NO: 112 Ac-LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys
Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu Lys;
SEQ ID NO: 113 Ac-LEU GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys
Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys;
SEQ ID NO: 114 Ac-LEU GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys;
SEQ ID NO: 115 SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu
Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr
Thr Lys Lys Leu Asn Thr GIn-NH2;
SEQ ID NO: 116 SER Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr
Thr Lys Lys Leu Asn Thr GIn-NH2;
SEQ ID NO: 117 SER Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn
Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LyS-NH2;
SEQ ID NO: 118 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe
Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp
Trp TIp-NH2; SEQ ID NO: 119 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn-NH2; SEQ ID NO: 120 SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp-NH2;
SEQ ID NO: 121 SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn-NH2; SEQ ID NO: 122 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp-NH2; SEQ ID NO: 123 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn-NH2; SEQ ID NO: 124 SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp-NH2; SEQ ID NO: 125 LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LyS-NH2; SEQ ID NO: 126 LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LyS-NH2; SEQ ID NO: 127 LEU GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys Pro Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu LyS-NH2;
SEQ ID NO: 128 LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LyS-NH2; SEQ ID NO: 129 LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LyS-NH2; SEQ ID NO: 130 LEU GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LyS-NH2; SEQ ID NO: 131 LEU GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LyS-NH2; SEQ ID NO: 132 Ac-SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala
Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu
Tyr Thr Lys Lys Leu Asn Thr GIn-NH2;
SEQ ID NO: 133 Ac-SER Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu
Tyr Thr Lys Lys Leu Asn Thr GIn-NH2;
SEQ ID NO: 134 Ac-SER Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu
Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LyS-NH2;
SEQ ID NO: 135 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe
Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Tip Arg Tip Tip
Tip Trp-NH2;
SEQ ID NO: 136 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met
Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn-NH2;
SEQ ID NO: 137 Ac-SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala
Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu
Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Tip Tip Tip Trp-NH2;
SEQ ID NO: 138 Ac-SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr
Thr Lys Lys Leu Asn Thr GIn-NH2;
SEQ ID NO: 139 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala
Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe
Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp Trp-NH2;
SEQ ID NO: 140 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala
Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe
Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GIn-NH2;
SEQ ID NO: 141 Ac-SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr
Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Tip Trp Trp Trp-NH2;
SEQ ID NO: 142 Ac-LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys
Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu Lys-
NH2; SEQ ID NO: 143 Ac-LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys
Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LyS-NH2;
SEQ ID NO: 144 Ac-LEU GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys
Pro Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu LyS-NH2; SEQ ID NO: 145 Ac-LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys
Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu Lys-
NH2;
SEQ ID NO: 146 Ac-LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys
Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LyS-NH2; SEQ ID NO: 147 Ac-LEU GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys
Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys-
NH2;
SEQ ID NO: 148 Ac-LEU GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys
Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LyS-NH2; SEQ ID NO: 149 ASP Tip Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe Pro
Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala Ala;
SEQ ID NO: 150 ALA Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala Phe;
SEQ ID NO: 151 PRO VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys
GIn Lys Leu Lys; SEQ ID NO: 152 Ac-ASP Trp Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe
Pro Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala Ala;
SEQ ID NO: 153 Ac-ALA Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala
Phe;
SEQ ID NO: 154 Ac-PRO VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys GIn Lys Leu Lys;
SEQ ID NO: 155 ASP Trp Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe Pro
Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala AIa-NH2;
SEQ ID NO: 156 ALA Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala Phe-
NH2; SEQ ID NO: 157 PRO VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys
GIn Lys Leu LyS-NH2;
SEQ ID NO: 158 Ac-ASP Trp Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe
Pro Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala AIa-NH2; SEQ ID NO: 159 Ac-ALA Tip Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala Phe-NH2;
SEQ ID NO: 160 Ac-PRO VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys GIn Lys Leu LyS-NH2.
vi. Peptides Containing a Carboxy-Terminal D Amino Acid
SEQ ID NO: 161 Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg
Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN;
SEQ ID NO: 162 Ser Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 163 Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS;
SEQ ID NO: 164 Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Tip TRP; SEQ ID NO: 165 Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 166 Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP;
SEQ ID NO: 167 Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 168 Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP; SEQ ID NO: 169 Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys
VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys
VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN;
SEQ ID NO: 170 Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr
Lys Lys Ala Leu Ser Pro Leu Trp Arg Tip Tip Tip TRP;
SEQ ID NO: 171 Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys
Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LYS;
SEQ ID NO: 172 Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LYS;
SEQ ID NO: 173 Leu GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys Pro
Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu LYS;
SEQ ID NO: 174 Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys
Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS; SEQ ID NO: 175 Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro
Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS;
SEQ ID NO: 176 Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys
Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS;
SEQ ID NO: 177 Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS;
SEQ ID NO: 178 Ac-Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu
Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr
Thr Lys Lys Leu Asn Thr GLN;
SEQ ID NO: 179 Ac-Ser Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr
Thr Lys Lys Leu Asn Thr GLN;
SEQ ID NO: 180 Ac-Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn
Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS;
SEQ ID NO: 181 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe
Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Tip Arg Tip Trp
Trp TRP; SEQ ID NO: 182 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Ly s GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 183 Ac-Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP;
SEQ ID NO: 184 Ac-Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 185 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP; SEQ ID NO: 186 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 187 Ac-Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP; SEQ ID NO: 188 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LYS; SEQ ID NO: 189 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LYS; SEQ ID NO: 190 Ac-Leu GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys Pro Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu LYS;
SEQ ID NO: 191 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS; SEQ ID NO: 192 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS; SEQ ID NO: 193 Ac-Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS; SEQ ID NO: 194 Ac-Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS; SEQ ID NO: 195 Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2;
SEQ ID NO: 196 Ser Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2;
SEQ ID NO: 197 Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS-NH2; SEQ ID NO: 198 Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Tip Trp TRP-NH2;
SEQ ID NO: 199 Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2;
SEQ ID NO: 200 Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP-NH2; SEQ ID NO: 201 Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2;
SEQ ID NO: 202 Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP-NH2;
SEQ ID NO: 203 Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2; SEQ ID NO: 204 Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP-NH2;
SEQ ID NO: 205 Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LYS-NH2; SEQ ID NO: 206 Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Ly s Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LYS-NH2; SEQ ID NO: 207 Leu GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys Pro Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu LYS-NH2; SEQ ID NO: 208 Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS-NH2; SEQ ID NO: 209 Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS-NH2; SEQ ID NO: 210 Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS-NH2; SEQ ID NO: 211 Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS-NH2; SEQ ID NO: 212 Ac-Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2;
SEQ ID NO: 213 Ac-Ser Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2; SEQ ID NO: 214 Ac-Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS-NH2;
SEQ ID NO: 215 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP-NH2; SEQ ID NO: 216 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2; SEQ ID NO: 217 Ac-Ser Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP-NH2;
SEQ ID NO: 218 Ac-Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2; SEQ ID NO: 219 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Tip Arg Tip Trp Tip TRP-NH2; SEQ ID NO: 220 Ac-Pro Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2; SEQ ID NO: 221 Ac-Ser Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP-NH2;
SEQ ID NO: 222 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys
Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LYS-
NH2;
SEQ ID NO: 223 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LYS-NH2;
SEQ ID NO: 224 Ac-Leu GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys Pro Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu LYS-NH2; SEQ ID NO: 225 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS- NH2;
SEQ ID NO: 226 Ac-Leu GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS-NH2; SEQ ID NO: 227 Ac-Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS- NH2;
SEQ ID NO: 228 Ac-Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS-NH2; SEQ ID NO: 229 Asp Trp Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe Pro Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala ALA; SEQ ID NO: 230 Ala Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala PHE;
SEQ ID NO: 231 Pro VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys GIn Lys Leu LYS;
SEQ ID NO: 232 Ac-Asp Trp Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe Pro Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala ALA; SEQ ID NO: 233 Ac-Ala Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala PHE;
SEQ ID NO: 234 Ac-Pro VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys
GIn Lys Leu LYS;
SEQ ID NO: 235 Asp Trp Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe Pro Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala ALA-NH2;
SEQ ID NO: 236 Ala Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala PHE-
NH2;
SEQ ID NO: 237 Pro VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys
GIn Lys Leu LYS-NH2. SEQ ID NO: 238 Ac-Asp Trp Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe
Pro Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala ALA-NH2;
SEQ ID NO: 239 Ac-Ala Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala PHE-
NH2;
SEQ ID NO: 240 Ac-Pro VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys GIn Lys Leu LYS-NH2.
vii. Peptides Containing Both Amino-Terminal and Carboxy-Terminal D Amino Acids
SEQ ID NO: 241 SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu
Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN;
SEQ ID NO: 242 SER Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 243 SER Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS;
SEQ ID NO: 244 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP; SEQ ID NO: 245 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 246 SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Tip TRP;
SEQ ID NO: 247 SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN;
SEQ ID NO: 248 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP;
SEQ ID NO: 249 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 250 SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP;
SEQ ID NO: 251 LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LYS; SEQ ID NO: 252 LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LYS;
SEQ ID NO: 253 LEU GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys Pro Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu LYS; SEQ ID NO: 254 LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS; SEQ ID NO: 255 LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS; SEQ ID NO: 256 LEU GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS; SEQ ID NO: 257 LEU GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS;
SEQ ID NO: 258 Ac-SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 259 Ac-SER Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN;
SEQ ID NO: 260 Ac-SER Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS;
SEQ ID NO: 261 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Tip Trp Trp TRP; SEQ ID NO: 262 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 263 Ac-SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP;
SEQ ID NO: 264 Ac-SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN; SEQ ID NO: 265 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP;
SEQ ID NO: 266 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN;
SEQ ID NO: 267 Ac-SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP; SEQ ID NO: 268 Ac-LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LYS; SEQ ID NO: 269 Ac-LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LYS; SEQ ID NO: 270 Ac-LEU GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys Pro Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu LYS; SEQ ID NO: 271 Ac-LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS; SEQ ID NO: 272 Ac-LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS; SEQ ID NO: 273 Ac-LEU GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS; SEQ ID NO: 274 Ac-LEU GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS; SEQ ID NO: 275 SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2;
SEQ ID NO: 276 SER Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2; SEQ ID NO: 277 SER Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS-NH2; SEQ ID NO: 278 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Tip Arg Trp Trp Trp TRP-NH2;
SEQ ID NO: 279 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2; SEQ ID NO: 280 SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP-NH2;
SEQ ID NO: 281 SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2; SEQ ID NO: 282 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP-NH2; SEQ ID NO: 283 PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys
VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys
VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2;
SEQ ID NO: 284 SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr
Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Tip Tip TRP-NH2;
SEQ ID NO: 285 LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys
Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LYS-NH2;
SEQ ID NO: 286 LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LYS-NH2;
SEQ ID NO: 287 LEU GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys Pro
Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu LYS-NH2;
SEQ ID NO: 288 LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Lys
Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS-NH2; SEQ ID NO: 289 LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys Pro
Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS-NH2;
SEQ ID NO: 290 LEU GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys
Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS-NH2;
SEQ ID NO: 291 LEU GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS-NH2;
SEQ ID NO: 292 Ac-SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala
Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu
Tyr Thr Lys Lys Leu Asn Thr GLN-NH2;
SEQ ID NO: 293 Ac-SER Pro Leu Ser Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu Asn Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu
Tyr Thr Lys Lys Leu Asn Thr GLN-NH2;
SEQ ID NO: 294 Ac-SER Asp GIu Leu Arg GIn Arg Leu Ala Ala Arg Leu GIu Ala Leu Lys GIu
Asn Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS-NH2;
SEQ ID NO: 295 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe
Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp
Trp TRP-NH2; SEQ ID NO: 296 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys GIy GIu GIu Met
Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe
Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2;
SEQ ID NO: 297 Ac-SER Pro Leu GIy GIu GIu Met Arg Asp Arg Ala Arg Ala His VaI Asp Ala Leu Arg Thr His Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu
Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP-NH2;
SEQ ID NO: 298 Ac-SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala
Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr
Thr Lys Lys Leu Asn Thr GLN-NH2; SEQ ID NO: 299 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala
Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe
Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp
Trp TRP-NH2;
SEQ ID NO: 300 Ac-PRO Arg GIy GIy Ser VaI Leu VaI Thr Leu Pro Ser Leu Lys Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe
Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys Lys Leu Asn Thr GLN-NH2;
SEQ ID NO: 301 Ac-SER Pro Leu Leu GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala
Thr Lys Lys Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr
Thr Lys Lys Ala Leu Ser Pro Leu Trp Arg Trp Trp Trp TRP-NH2; SEQ ID NO: 302 Ac-LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys
Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LYS-
NH2;
SEQ ID NO: 303 Ac-LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys
Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Ala Ala GIu LYS-NH2; SEQ ID NO: 304 Ac-LEU GIu Ser Ala Phe VaI Ser Leu Lys Ser Phe Tyr GIu GIu VaI Ala GIu Lys
Pro Ala GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Tyr GIu GIu Ala Ala GIu LYS-NH2;
SEQ ID NO: 305 Ac-LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys
Lys Leu Ser Pro Leu Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS-
NH2; SEQ ID NO: 306 Ac-LEU GIu Ser Phe Leu VaI Ser Phe Lys Ser Ala Leu GIu GIu Tyr Phe GIu Lys
Pro Leu GIu Ser Ala Ala VaI Ser Ala Lys Ser Ala Leu GIu GIu Tyr Ala GIu LYS-NH2;
SEQ ID NO: 307 Ac-LEU GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys
Lys Leu Ser Pro Leu Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS-
NH2; SEQ ID NO: 308 Ac-LEU GIu Ser Ala Lys VaI Ser Ala Leu Ser Ala Leu GIu GIu Ala Thr Lys Lys
Pro Leu GIu Ser Phe Lys VaI Ser Phe Leu Ser Ala Leu GIu GIu Tyr Thr Lys LYS-NH2;
SEQ ID NO: 309 ASP Trp Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe Pro
Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala ALA; SEQ ID NO: 310 ALA Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala PHE;
SEQ ID NO: 311 PRO VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys
GIn Lys Leu LYS;
SEQ ID NO: 312 Ac-ASP Trp Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe
Pro Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala ALA; SEQ ID NO: 313 Ac-ALA Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala
PHE;
SEQ ID NO: 314 Ac-PRO VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu
Lys GIn Lys Leu LYS;
SEQ ID NO: 315 ASP Trp Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe Pro Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala ALA-NH2;
SEQ ID NO: 316 ALA Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala PHE-
NH2;
SEQ ID NO: 317 PRO VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys
GIn Lys Leu LYS-NH2. SEQ ID NO: 318 Ac-ASP Trp Leu Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Leu Lys GIu Ala Phe
Pro Asp Trp Ala Lys Ala Ala Tyr Asp Lys Ala Ala GIu Lys Ala Lys GIu Ala ALA-NH2;
SEQ ID NO: 319 Ac-ALA Trp Phe Lys Ala Phe Tyr Asp Lys VaI Ala GIu Lys Phe Lys GIu Ala
PHE-NH2;
SEQ ID NO: 320 Ac-PRO VaI Leu Asp Leu Phe Arg GIu Leu Leu Asn GIu Leu Leu GIu Ala Leu Lys GIn Lys Leu LYS-NH2.
viii. Peptides Containing AU D Amino Acids
SEQ ID NO: 321 SER PRO LEU GLY GLU GLU MET ARG ASP ARG ALA ARG ALA HIS VAL ASP ALA LEU ARG THRHIS LYS LEU SERPRO LEU LEU GLU SERPHE LYS VAL SERPHELEU SERALALEUGLU GLUTYRTHRLYS LYS LEUASNTHRGLN;
SEQ ID NO: 322 SER PRO LEU SER ASP GLU LEU ARG GLN ARG LEU ALA ALA ARG LEU GLU ALALEU LYS GLU ASN LYS LEU SERPRO LEU LEU GLU SERPHE LYS VAL SERPHELEU SERALALEUGLUGLUTYRTHRLYS LYS LEUASNTHRGLN; SEQ ID NO: 323 SER ASP GLU LEU ARG GLN ARG LEU ALA ALA ARG LEU GLU ALA LEU LYS GLU ASN PRO LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS;
SEQ ID NO: 324 PRO ARG GLY GLY SER VAL LEU VAL THR LEU PRO SER LEU LYS GLY GLU GLU MET ARG ASP ARG ALA ARG ALA HIS VAL ASP ALA LEU ARG THR HIS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS ALA LEU SER PRO LEU TRP ARG TRP TRP TRP TRP; SEQ ID NO: 325 PRO ARG GLY GLY SER VAL LEU VAL THR LEU PRO SER LEU LYS GLY GLU GLU MET ARG ASP ARG ALA ARG ALA HIS VAL ASP ALA LEU ARG THR HIS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN;
SEQ ID NO: 326 SER PRO LEU GLY GLU GLU MET ARG ASP ARG ALA ARG ALA HIS VAL ASP ALA LEU ARG THR HIS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS ALA LEU SER PRO LEU TRP ARG TRP TRP TRP TRP;
SEQ ID NO: 327 SER PRO LEU LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN; SEQ ID NO: 328 PRO ARG GLY GLY SER VAL LEU VAL THR LEU PRO SER LEU LYS LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS ALA LEU SER PRO LEU TRP ARG TRP TRP TRP TRP; SEQ ID NO: 329 PRO ARG GLY GLY SER VAL LEU VAL THR LEU PRO SER LEU LYS LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN;
SEQ ID NO: 330 SER PRO LEU LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS ALA LEU SER PRO LEU TRP ARG TRP TRP TRP TRP;
SEQIDNO: 331 LEUGLU SERPHELEUVAL SERPHELYS SERALALEU GLU GLUTYR PHE GLU LYS LYS LEU SERPRO LEU LEU GLU SERALA ALAVAL SERALA LYS SER ALALEUGLUGLUALAALAGLULYS; SEQIDNO: 332 LEUGLUSERPHELEUVAL SERPHELYS SERALALEUGLU GLUTYR
PHEGLULYSPROLEUGLU SERALAALAVAL SERALALYS SERALALEUGLUGLU
ALAALAGLULYS;
SEQIDNO: 333 LEUGLUSERALAPHEVAL SERLEULYS SERPHETYRGLUGLUVAL ALAGLULYS PROALAGLU SERALAALAVAL SERALALYS SERALATYRGLUGLU
ALAALAGLULYS;
SEQIDNO: 334LEUGLUSERPHELEUVALSERPHELYS SERALALEUGLUGLUTYR
PHE GLU LYS LYS LEU SERPRO LEU LEU GLU SERALA ALAVAL SERALA LYS SER
ALALEUGLUGLUTYRALAGLULYS; SEQIDNO: 335 LEUGLU SERPHELEUVAL SERPHELYS SERALALEUGLU GLUTYR
PHE GLULYS PRO LEU GLU SERALAALAVAL SERALALYS SERALALEU GLUGLU
TYRALAGLULYS;
SEQ ID NO: 336 LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU
ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SERPHE LEU SERALALEUGLUGLUTYRTHRLYS LYS;
SEQ ID NO: 337 LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU
ALA THRLYS LYS PRO LEU GLU SERPHELYS VAL SERPHELEU SERALALEU GLU
GLUTYRTHRLYS LYS;
SEQ ID NO: 338 Ac-SER PRO LEU GLY GLU GLU MET ARG ASP ARG ALA ARG ALA HIS VAL ASP ALA LEU ARG THR HIS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL
SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN;
SEQ ID NO: 339 Ac-SER PRO LEU SER ASP GLU LEU ARG GLN ARG LEU ALA ALA ARG
LEU GLU ALA LEU LYS GLU ASN LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL
SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN; SEQ ID NO: 340 Ac-SER ASP GLU LEU ARG GLN ARG LEU ALA ALA ARG LEU GLU ALA
LEU LYS GLU ASN PRO LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU
GLU TYR THR LYS LYS;
SEQ ID NO: 341 AC-PRO ARG GLY GLY SER VAL LEU VAL THR LEU PRO SER LEU LYS
GLY GLU GLU MET ARG ASP ARG ALA ARG ALA HIS VAL ASP ALA LEU ARG THR HIS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU
GLU TYR THR LYS LYS ALA LEU SER PRO LEU TRP ARG TRP TRP TRP TRP;
SEQ ID NO: 342 Ac-PRO ARG GLY GLY SER VAL LEU VAL THR LEU PRO SER LEU LYS
GLY GLU GLU MET ARG ASP ARG ALA ARG ALA HIS VAL ASP ALA LEU ARG THR HIS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN;
SEQ ID NO: 343 Ac-SER PRO LEU GLY GLU GLU MET ARG ASP ARG ALA ARG ALA HIS VAL ASP ALA LEU ARG THR HIS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS ALA LEU SER PRO LEU TRP ARG TRP TRP TRP TRP;
SEQ ID NO: 344 Ac-SER PRO LEU LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN; SEQ ID NO: 345 Ac-PRO ARG GLY GLY SER VAL LEU VAL THR LEU PRO SER LEU LYS LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS ALA LEU SER PRO LEU TRP ARG TRP TRP TRP TRP; SEQ ID NO: 346 Ac-PRO ARG GLY GLY SER VAL LEU VAL THR LEU PRO SER LEU LYS LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN;
SEQ ID NO: 347 Ac-SER PRO LEU LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS ALA LEU SER PRO LEU TRP ARG TRP TRP TRP TRP;
SEQ ID NO: 348 Ac-LEU GLU SER PHE LEU VAL SER PHE LYS SER ALA LEU GLU GLU TYR PHE GLU LYS LYS LEU SER PRO LEU LEU GLU SER ALA ALA VAL SER ALA LYS SER ALA LEU GLU GLU ALA ALA GLU LYS; SEQ ID NO: 349 Ac-LEU GLU SER PHE LEU VAL SER PHE LYS SER ALA LEU GLU GLU TYR PHE GLU LYS PRO LEU GLU SER ALA ALA VAL SER ALA LYS SER ALA LEU GLU GLU ALA ALA GLU LYS;
SEQ ID NO: 350 Ac-LEU GLU SER ALA PHE VAL SER LEU LYS SER PHE TYR GLU GLU VAL ALA GLU LYS PRO ALA GLU SER ALA ALA VAL SER ALA LYS SER ALA TYR GLU GLU ALA ALA GLU LYS;
SEQ ID NO: 351 Ac-LEU GLU SER PHE LEU VAL SER PHE LYS SER ALA LEU GLU GLU TYR PHE GLU LYS LYS LEU SER PRO LEU LEU GLU SER ALA ALA VAL SER ALA LYS SER ALA LEU GLU GLU TYR ALA GLU LYS; SEQ ID NO: 352 Ac-LEU GLU SER PHE LEU VAL SER PHE LYS SER ALA LEU GLU GLU TYR PHE GLU LYS PRO LEU GLU SER ALA ALA VAL SER ALA LYS SER ALA LEU GLU GLU TYR ALA GLU LYS;
SEQ ID NO: 353 Ac-LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS;
SEQ ID NO: 354 Ac-LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS PRO LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS; SEQ ID NO: 355 SER PRO LEU GLY GLU GLU MET ARG ASP ARG ALA ARG ALA HIS VAL ASP ALA LEU ARG THR HIS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN-NH2; SEQ ID NO: 356 SER PRO LEU SER ASP GLU LEU ARG GLN ARG LEU ALA ALA ARG LEU GLU ALA LEU LYS GLU ASN LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN-NH2;
SEQ ID NO: 357 SER ASP GLU LEU ARG GLN ARG LEU ALA ALA ARG LEU GLU ALA LEU LYS GLU ASN PRO LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS-NH2; SEQ ID NO: 358 PRO ARG GLY GLY SER VAL LEU VAL THR LEU PRO SER LEU LYS GLY GLU GLU MET ARG ASP ARG ALA ARG ALA HIS VAL ASP ALA LEU ARG THR HIS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS ALA LEU SER PRO LEU TRP ARG TRP TRP TRP TRP-NH2; SEQ ID NO: 359 PRO ARG GLY GLY SER VAL LEU VAL THR LEU PRO SER LEU LYS GLY GLU GLU MET ARG ASP ARG ALA ARG ALA HIS VAL ASP ALA LEU ARG THR HIS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN-NH2;
SEQ ID NO: 360 SER PRO LEU GLY GLU GLU MET ARG ASP ARG ALA ARG ALA HIS VAL ASP ALA LEU ARG THR HIS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS ALA LEU SER PRO LEU TRP ARG TRP TRP TRP TRP-NH2;
SEQ ID NO: 361 SER PRO LEU LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN-NH2; SEQ ID NO: 362 PRO ARG GLY GLY SER VAL LEU VAL THR LEU PRO SER LEU LYS LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS ALA LEU SER PRO LEU TRP ARG TRP TRP TRP TRP-NH2; SEQ ID NO: 363 PRO ARG GLY GLY SER VAL LEU VAL THR LEU PRO SER LEU LYS LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN-NH2; SEQ ID NO: 364 SER PRO LEU LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS ALA LEU SER PRO LEU TRP ARG TRP TRP TRP TRP-NH2;
SEQ ID NO: 365 LEU GLU SER PHE LEU VAL SER PHE LYS SER ALA LEU GLU GLU TYR PHE GLU LYS LYS LEU SER PRO LEU LEU GLU SER ALA ALA VAL SER ALA LYS SER ALA LEU GLU GLU ALA ALA GLU LYS-NH2;
SEQ ID NO: 366 LEU GLU SER PHE LEU VAL SER PHE LYS SER ALA LEU GLU GLU TYR PHE GLU LYS PRO LEU GLU SER ALA ALA VAL SER ALA LYS SER ALA LEU GLU GLU ALA ALA GLU LYS-NH2; SEQ ID NO: 367 LEU GLU SER ALA PHE VAL SER LEU LYS SER PHE TYR GLU GLU VAL ALA GLU LYS PRO ALA GLU SER ALA ALA VAL SER ALA LYS SER ALA TYR GLU GLU ALA ALA GLU LYS-NH2;
SEQ ID NO: 368 LEU GLU SER PHE LEU VAL SER PHE LYS SER ALA LEU GLU GLU TYR PHE GLU LYS LYS LEU SER PRO LEU LEU GLU SER ALA ALA VAL SER ALA LYS SER ALA LEU GLU GLU TYR ALA GLU LYS-NH2; SEQ ID NO: 369 LEU GLU SER PHE LEU VAL SER PHE LYS SER ALA LEU GLU GLU TYR PHE GLU LYS PRO LEU GLU SER ALA ALA VAL SER ALA LYS SER ALA LEU GLU GLU TYR ALA GLU LYS-NH2;
SEQ ID NO: 370 LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS-NH2;
SEQ ID NO: 371 LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS PRO LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS-NH2; SEQ ID NO: 372 Ac-SER PRO LEU GLY GLU GLU MET ARG ASP ARG ALA ARG ALA HIS VAL ASP ALA LEU ARG THR HIS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN-NH2; SEQ ID NO: 373 Ac-SER PRO LEU SER ASP GLU LEU ARG GLN ARG LEU ALA ALA ARG LEU GLU ALA LEU LYS GLU ASN LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN-NH2; SEQ ID NO: 374 Ac-SER ASP GLU LEU ARG GLN ARG LEU ALA ALA ARG LEU GLU ALA LEU LYS GLU ASN PRO LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS-NH2; SEQ ID NO: 375 Ac-PRO ARG GLY GLY SER VAL LEU VAL THR LEU PRO SER LEU LYS GLY GLU GLU MET ARG ASP ARG ALA ARG ALA HIS VAL ASP ALA LEU ARG THR HIS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS ALA LEU SER PRO LEU TRP ARG TRP TRP TRP TRP-NH2; SEQ ID NO: 376 Ac-PRO ARG GLY GLY SER VAL LEU VAL THR LEU PRO SER LEU LYS GLY GLU GLU MET ARG ASP ARG ALA ARG ALA HIS VAL ASP ALA LEU ARG THR HIS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN-NH2;
SEQ ID NO: 377 Ac-SER PRO LEU GLY GLU GLU MET ARG ASP ARG ALA ARG ALA HIS VAL ASP ALA LEU ARG THR HIS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS ALA LEU SER PRO LEU TRP ARG TRP TRP TRP TRP-NH2;
SEQ ID NO: 378 Ac-SER PRO LEU LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN-NH2; SEQ ID NO: 379 Ac-PRO ARG GLY GLY SER VAL LEU VAL THR LEU PRO SER LEU LYS LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS ALA LEU SER PRO LEU TRP ARG TRP TRP TRP TRP-NH2; SEQ ID NO: 380 Ac-PRO ARG GLY GLY SER VAL LEU VAL THR LEU PRO SER LEU LYS LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS LEU ASN THR GLN-NH2;
SEQ ID NO: 381 Ac-SER PRO LEU LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SERPHE LEU SERALALEU GLU GLU TYRTHRLYS LYS ALA LEU SERPRO LEU TRP
ARGTRPTRPTRPTRP-NH2;
SEQ ID NO: 382 Ac-LEU GLU SER PHE LEU VAL SER PHE LYS SER ALA LEU GLU GLU
TYR PHE GLU LYS LYS LEU SER PRO LEU LEU GLU SER ALA ALA VAL SER ALA LYS SER ALA LEU GLU GLU ALA ALA GLU LYS-NH2;
SEQ ID NO: 383 Ac-LEU GLU SER PHE LEU VAL SER PHE LYS SER ALA LEU GLU GLU
TYR PHE GLU LYS PRO LEU GLU SER ALA ALA VAL SER ALA LYS SER ALA LEU GLU
GLU ALA ALA GLU LYS-NH2;
SEQ ID NO: 384 Ac-LEU GLU SER ALA PHE VAL SER LEU LYS SER PHE TYR GLU GLU VAL ALA GLU LYS PRO ALA GLU SER ALA ALA VAL SER ALA LYS SER ALA TYR GLU
GLU ALA ALA GLU LYS-NH2;
SEQ ID NO: 385 Ac-LEU GLU SER PHE LEU VAL SER PHE LYS SER ALA LEU GLU GLU
TYR PHE GLU LYS LYS LEU SER PRO LEU LEU GLU SER ALA ALA VAL SER ALA LYS
SER ALA LEU GLU GLU TYR ALA GLU LYS-NH2; SEQ ID NO: 386 Ac-LEU GLU SER PHE LEU VAL SER PHE LYS SER ALA LEU GLU GLU
TYR PHE GLU LYS PRO LEU GLU SER ALA ALA VAL SER ALA LYS SER ALA LEU GLU
GLU TYR ALA GLU LYS-NH2;
SEQ ID NO: 387 Ac-LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU
ALA THR LYS LYS LYS LEU SER PRO LEU LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU GLU TYR THR LYS LYS-NH2;
SEQ ID NO: 388 Ac-LEU GLU SER ALA LYS VAL SER ALA LEU SER ALA LEU GLU GLU
ALA THR LYS LYS PRO LEU GLU SER PHE LYS VAL SER PHE LEU SER ALA LEU GLU
GLU TYR THR LYS LYS-NH2;
SEQIDNO: 389ASP TRPLEU LYS ALAPHETYRASP LYS VALALA GLU LYS LEU LYS GLUALAPHEPROASP TRP ALALYS ALAALATYRASP LYS ALAALAGLU LYS ALA
LYSGLUALAALA;
SEQIDNO: 390ALATRPPHELYSALAPHETYRASPLYS VALALAGLULYSPHELYS
GLUALAPHE;
SEQ ID NO: 391 PRO VAL LEU ASP LEU PHE ARG GLU LEU LEU ASN GLU LEU LEU GLU ALA LEU LYS GLN LYS LEU LYS;
SEQ ID NO: 392 Ac-ASP TRP LEU LYS ALA PHE TYR ASP LYS VAL ALA GLU LYS LEU
LYS GLU ALA PHE PRO ASP TRP ALA LYS ALA ALA TYR ASP LYS ALA ALA GLU LYS
ALA LYS GLU ALA ALA; SEQ ID NO: 393 Ac-ALA TRP PHE LYS ALA PHE TYR ASP LYS VAL ALA GLU LYS PHE
LYS GLU ALA PHE;
SEQ ID NO: 394 Ac-PRO VAL LEU ASP LEU PHE ARG GLU LEU LEU ASN GLU LEU LEU
GLU ALA LEU LYS GLN LYS LEU LYS; SEQ ID NO: 395 ASP TRP LEU LYS ALA PHE TYR ASP LYS VAL ALA GLU LYS LEU LYS
GLU ALA PHE PRO ASP TRP ALA LYS ALA ALA TYR ASP LYS ALA ALA GLU LYS ALA
LYS GLU ALA ALA-NH2;
SEQ ID NO: 396 ALA TRP PHE LYS ALA PHE TYR ASP LYS VAL ALA GLU LYS PHE LYS
GLU ALA PHE-NH2; SEQ ID NO: 397 PRO VAL LEU ASP LEU PHE ARG GLU LEU LEU ASN GLU LEU LEU
GLU ALA LEU LYS GLN LYS LEU LYS-NH2.
SEQ ID NO: 398 Ac-ASP TRP LEU LYS ALA PHE TYR ASP LYS VAL ALA GLU LYS LEU
LYS GLU ALA PHE PRO ASP TRP ALA LYS ALA ALA TYR ASP LYS ALA ALA GLU LYS
ALA LYS GLU ALA ALA-NH2; SEQ ID NO: 399 Ac-ALA TRP PHE LYS ALA PHE TYR ASP LYS VAL ALA GLU LYS PHE
LYS GLU ALA PHE-NH2;
SEQ IDNO: 400 Ac-PRO VAL LEU ASP LEU PHEARG GLU LEU LEUASN GLU LEU LEU
GLUALALEULYS GLNLYS LEULYS-NH2.
C. Peptide/Lipid Complexes
The HDL mimetic peptides of the present invention may be complexed with one or more lipids, e.g., one or more electrically neutral and/or electrically charged phospholipids. Particularly for oral administration, these peptide/lipid complexes may then be encapsulated into vesicles or nanoparticles as described herein. In one example, peptides are reconstituted with one or more lipids prior to encapsulating the peptide/lipid complexes within vesicles or nanoparticles. The peptides may be reconstituted with an electrically neutral phospholipid (i.e., a phospholipid having zero net electrical charge), e.g., a lipid containing phospatidylcholine (lecithin) or sphingomyelin or a mixture thereof. The peptides may also be reconstituted with an electrically charged phospholipid, e.g., but not limited to, a lipid containing phosphatidylserine, phosphatidylinositol, phosphatidylglycerol, l,2-diacylglycerol-3-phosphate (phosphatidic acid), and/or a mixture thereof (each of these phospholipids has a net electrical charge of minus one). In some embodiments the peptide is reconstituted with a neutral phospholipid plus a charged phospholipid. Such charged lipoprotein complexes containing an HDL mimetic peptide or any full-length apoplipoprotein such as but not limited to ApoA-I (see e.g., US 20060217312) can be produced using well-known methods, e.g. but not limited to the methods set forth in US 20060217312 or US 7,189,411. Such HDL mimetic peptide/lipid complexes or apolipoprotein/lipid complexes can then be encapsulated into vesicles or nanoparticles as described herein.
Some examples of neutral phospholipids containing phosphatidylcholine that may be employed to produce peptide/lipid complexes include but are not limited to the following: dipalmitoylphosphatidylcholine (DPPC); dioleoylphosphatidylcholine (DOPC); l-palmitoyl-2- oleoylphosphatidylcholine (POPC); l-palmitoyl-2-linoleoylphosphatidylcholine (PLPC); 1- palmitoyl-2-arachidonylphosphatidylcholine (PAPC); l-palmitoyl-2- docosahexanoylphosphatidylcholine (PDPC); and l-palmitoyl-2-myristoylphosphatidylcholine (PMPC). See, e.g., Shah et al. (Circulation (2001), 103(25):3047-50), in which DPPC was used to reconstitute peptides. One of ordinary skill in the art will understand that phospholipids containing different types of fatty acids, e.g. but not limited to palmitic acid, stearic acid, myristic acid, oleic acid, linoleic acid, linolenic acid, and/or any other type of fatty acid (see, e.g., Chapter 5 of Zubay G., Biochemistry 2nd ed., MacMillan Publishing Co., New York, 1988 or any other biochemistry textbook), can be used in the peptide/lipid complexes and peptide/apolipoprotein complexes of the present invention.
The peptides of the present invention may be complexed with neutral and/or charged phospholipids or any mixture thereof in molar ratios ranging from about 1:0.5 to about 1:200. For example the peptide to lipid ratio can be, but is not limited to, 1:0.5 to 1:10; 1:1 to 1: 5; 1:2 to 1:10; 1 : 1 to 1 :20; 1 :40 to 1 : 100; 1 : 100.1 to 1 :200. Any ratio within these ranges may be employed. When charged phospholipids are mixed with neutral phospholipids within the present peptide/lipid complexes, the molar ratio of charged phospholipid to neutral phospholipid may be (but is not limited to), e.g., about 400:1 to about 1:400 (e.g., about 1:1 to 1:10, 1:5 to 1:20, 1:5 to 1:50, 1:10 to 1:100, 1:20 to 1:200, 1:40 to 1:400, 1:1 to 10:1, 5:1 to 20:1, 5:1 to 50:1, 10:1 to 100:1, 20:1 to 200:1, 40:1 to 400:1). Any ratio within these ranges may be employed in the present compositions. See, e.g., US 20060217312 for other appropriate ratios of peptide to phospholipid and charged phospholipid to uncharged phospholipid.
D. Modified HDL Mimetic Peptides
The present invention may be used for the production of the peptides or peptide analogs of the present invention. "Proteins", "peptides," "polypeptides" and "oligopeptides" are chains of amino acids (typically L-amino acids) whose alpha carbons are linked through peptide bonds formed by a condensation reaction between the carboxyl group of the alpha carbon of one amino acid and the amino group of the alpha carbon of another amino acid. The terminal amino acid at one end of the chain (i.e., the amino terminal) has a free amino group, while the terminal amino acid at the other end of the chain (i.e., the carboxy terminal) has a free carboxyl group. As such, the term "amino terminus" (abbreviated N-terminus) refers to the free alpha-amino group on the amino acid at the amino terminal of the protein, or to the alpha-amino group (imino group when participating in a peptide bond) of an amino acid at any other location within the protein. Similarly, the term "carboxy terminus" (abbreviated C-terminus) refers to the free carboxyl group on the amino acid at the carboxy terminus of a protein, or to the carboxyl group of an amino acid at any other location within the protein.
Typically, the amino acids making up a protein are numbered in order, starting at the amino terminal and increasing in the direction toward the carboxy terminal of the protein. Thus, when one amino acid is said to "follow" another, that amino acid is positioned closer to the carboxy terminal of the protein than the preceding amino acid.
The term "residue" is used herein to refer to an amino acid (D or L) or an amino acid mimetic that is incorporated into a protein by an amide bond. When a D amino acid is present in the peptides of the present invention, the three letter designation for the amino acid appears in upper case instead of a capital letter. For example the amino acid serine, represented as Ser indicates an L amino acid. The D amino acid form is represented as the upper case letters SER. This is not to be confused with letters appearing as subscripts used in generic formula and defined as variables herein. As such, the amino acid may be a naturally occurring amino acid or, unless otherwise limited, may encompass known analogs of natural amino acids that function in a manner similar to the naturally occurring amino acids (i.e., amino acid mimetics). Moreover, an amide bond mimetic includes peptide backbone modifications well known to those skilled in the art.
i. HDL Mimetic Peptides with Isosteric Amino Acid Substitutions and Substitutions Between Isosteric Amino Acid Groups
The present invention includes HDL mimetic peptides containing isosteric amino acid substitutions at specific amino acid positions within the peptides. By "isosteric substitution" is meant that an amino acid at a particular position within a peptide of the invention can be substituted with another amino acid belonging to the same isosteric group as described hereinbelow. Amino acids within a given isosteric group, as set forth hereinbelow, are amino acids having similar size, shape, polar/nonpolar properties, charge, and/or steric properties. The invention provides peptides wherein substitution of an amino acid with an amino acid belonging to the same isosteric group allows the substituted peptide to retain at least about 20% of the biological activity of the unsubstituted peptide, e.g., at least about: 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 110%, 125%, 150%, 175%, 200%, 250%, 300%, or more, of the biological activity of the unsubstituted peptide. By "biological activity" of the peptide is meant the ability of the peptide to promote lipid efflux and and/or have an anti-inflammatory effect, as described hereinbelow. The invention also provides peptides having an amino acid substituted with an amino acid belonging to a different isosteric group from the original amino acid, such that the substitution allows the peptide to retain at least about 20% of the biological activity (e.g., lipid efflux and/or anti- inflammatory properties) of the unsubstituted peptide, e.g., at least about: 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 110%, 125%, 150%, 175%, 200%, 250%, 300%, or more, of the biological activity of the unsubstituted peptide.
Peptides with an amino acid substitution that retain at least about 20% or more of the biological activity of an unsubstituted peptide will be readily recognized by the skilled artisan using well-known approaches, e.g., the lipid efflux and/or other biological assays of the invention. Isosteric amino acid groupings of the present invention are as follows: a) Isosteric Group 1 : Lys, His, and Arg; b) Isosteric Group 2: Asp and GIu; c) Isosteric Group 3: Ser, Thr, Leu, He, GIy, Ala, VaI, and GABA; d) Isosteric Group 4: Phe and Tyr; and e) Isosteric Group 5: Pro. An amino acid belonging to a given isosteric group can be substituted by a different amino acid within that same isosteric group.
In addition, an amino acid within a given isosteric group can be substituted by an amino acid from a different isosteric group as follows: a) an amino acid within Isosteric Group 1 can be substituted with an amino acid within
Isosteric Group 2; b) an amino acid within Isosteric Group 2 can be substituted with an amino acid within Isosteric Group 1, Isosteric Group 3, or Isosteric Group 4; c) an amino acid within Isosteric Group 3 can be substituted with an amino acid within Isosteric Group 4; moreover, an amino acid within Isosteric Group 3 that resides on a hydrophilic surface of an amphipathic helix of an HDL mimetic peptide of the invention can be substituted with an amino acid within either Isosteric Group 1 or Isosteric Group 2; d) An amino acid within Isosteric Group 4 can be substituted with an amino acid within Isosteric Group 3.
ii. HDL Mimetic Peptides with Conservative and Non-Conservative ("Severe"') Amino Acid Substitutions
One of skill will recognize that individual substitutions, deletions, or additions that alter, add, or delete a single amino acid or a small percentage of amino acids (typically fewer than about 5%, or typically fewer than about 1%) in a sequence are conservatively modified variations where the alterations result in the substitution of an amino acid with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art. The following six groups each contain amino acids that are conservative substitutions for one another: 1) Alanine (A), Serine (S), Threonine (T);
2) Aspartic acid (D), Glutamic acid (E);
3) Asparagine (N), Glutamine (Q), Histidine (H);
4) Arginine (R), Lysine (K);
5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V); and 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W).
A conservative substitution is a substitution in which the substituting amino acid (naturally occurring or modified) is structurally related to the amino acid being substituted, i.e., has about the same size and electronic properties as the amino acid being substituted. Thus, the substituting amino acid would have the same or a similar functional group in the side chain as the original amino acid. A "conservative substitution" also refers to utilizing a substituting amino acid which is identical to the amino acid being substituted except that a functional group in the side chain is protected with a suitable protecting group. Peptides of the present invention include conservatively substituted peptides, wherein these conservative substitutions occur at 1%, 3%, 5%, 7%, 10%, 15%, 20%, 25%, 30%, 40%, or 50% of the amino acid residues. Peptides of the present invention include peptides that are homologous at 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99% of the entire sequence of the peptide.
Suitable protecting groups are described in Green and Wuts, "Protecting Groups in Organic Synthesis", John Wiley and Sons, Chapters 5 and 7, 1991, the teachings of which are incorporated herein by reference. Preferred protecting groups are those which facilitate transport of the peptide through membranes, for example, by reducing the hydrophilicity and increasing the lipophilicity of the peptide, and which can be cleaved, either by hydrolysis or enzymatically (Ditter et al., 1968. J. Pharm. Sci. 57:783; Ditter et al., 1968. J. Pharm. Sci. 57:828; Ditter et al., 1969. J. Pharm. Sci. 58:557; King et al., 1987. Biochemistry 26:2294; Lindberg et al., 1989. Drug Metabolism and Disposition 17:311; Tunek et al., 1988. Biochem. Pharm. 37:3867; Anderson et al., 1985 Arch. Biochem. Biophys. 239:538; and Singhal et al., 1987. FASEB J. 1:220). Suitable hydroxyl protecting groups include ester, carbonate and carbamate protecting groups. Suitable amine protecting groups include acyl groups and alkoxy or aryloxy carbonyl groups, as described above for N-terminal protecting groups. Suitable carboxylic acid protecting groups include aliphatic, benzyl and aryl esters, as described below for C-terminal protecting groups. In one embodiment, the carboxylic acid group in the side chain of one or more glutamic acid or aspartic acid residues in a peptide of the present invention is protected, preferably as a methyl, ethyl, benzyl or substituted benzyl ester, more preferably as a benzyl ester.
Provided below are groups of naturally occurring and modified amino acids in which each amino acid in a group has similar electronic and steric properties. Thus, a conservative substitution can be made by substituting an amino acid with another amino acid from the same group. A "nonconservative substitution" can be made by substituting an amino acid with another amino acid from a different group. It is to be understood that these groups are non-limiting, i.e. that there are additional modified amino acids which could be included in each group. Group I includes leucine, isoleucine, valine, methionine and modified amino acids having the following side chains: ethyl, n-propyl n-butyl. Preferably, Group I includes leucine, isoleucine, valine and methionine.
Group II includes glycine, alanine, valine and a modified amino acid having an ethyl side chain. Preferably, Group II includes glycine and alanine. Group III includes phenylalanine, phenylglycine, tyrosine, tryptophan, cyclohexylmethyl glycine, and modified amino residues having substituted benzyl or phenyl side chains. Preferred substituents include one or more of the following: halogen, methyl, ethyl, nitro, — NH2, methoxy, ethoxy and — CN. Preferably, Group III includes phenylalanine, tyrosine and tryptophan.
Group IV includes glutamic acid, aspartic acid, a substituted or unsubstituted aliphatic, aromatic or benzylic ester of glutamic or aspartic acid (e.g., methyl, ethyl, n-propyl iso-propyl, cyclohexyl, benzyl or substituted benzyl), glutamine, asparagine, — CO — NH — alkylated glutamine or asparagines (e.g., methyl, ethyl, n-propyl and iso-propyl) and modified amino acids having the side chain — (CH2)3 — COOH, an ester thereof (substituted or unsubstituted aliphatic, aromatic or benzylic ester), an amide thereof and a substituted or unsubstituted N-alkylated amide thereof. Preferably, Group IV includes glutamic acid, aspartic acid, methyl aspartate, ethyl aspartate, benzyl aspartate and methyl glutamate, ethyl glutamate and benzyl glutamate, glutamine and asparagine. Group V includes histidine, lysine, ornithine, arginine, N-nitroarginine, β-cycloarginine, γ- hydroxyarginine, N-amidinocitruline and 2-amino-4-guanidinobutanoic acid, homologs of lysine, homologs of arginine and homologs of ornithine. Preferably, Group V includes histidine, lysine, arginine and ornithine. A homolog of an amino acid includes from 1 to about 3 additional or subtracted methylene units in the side chain. Group VI includes serine, threonine, and modified amino acids having C1-C5 straight or branched alkyl side chains substituted with — OH or — SH, for example, — CH2CH2OH, -CH2CH2CH2OH or -CH2CH2OHCH3. Preferably, Group VI includes serine, or threonine.
In another aspect, suitable substitutions for amino acid residues include "severe" or "non- conservative" substitutions. The terms "severe" or "non-conservative" substitution are used interchangeablt throughout this application. A "severe substitution" is a substitution in which the substituting amino acid (naturally occurring or modified) has significantly different size and/or electronic properties compared with the amino acid being substituted. Thus, the side chain of the substituting amino acid can be significantly larger (or smaller) than the side chain of the amino acid being substituted and/or can have functional groups with significantly different electronic properties than the amino acid being substituted. Examples of severe substitutions of this type include the substitution of phenylalanine or cyclohexylmethyl glycine for alanine, isoleucine for glycine, a D amino acid for the corresponding L amino acid, or — NH — CH[( — CH2)5 — COOH] — CO — for aspartic acid. Alternatively, a functional group may be added to the side chain, deleted from the side chain or exchanged with another functional group. Examples of severe substitutions of this type include adding of valine, leucine or isoleucine, exchanging the carboxylic acid in the side chain of aspartic acid or glutamic acid with an amine, or deleting the amine group in the side chain of lysine or ornithine, hi yet another alternative, the side chain of the substituting amino acid can have significantly different steric and electronic properties that the functional group of the amino acid being substituted. Examples of such modifications include tryptophan for glycine, lysine for aspartic acid and — (CH2)4COOH for the side chain of serine. These examples are not meant to be limiting.
In addition to the naturally occurring genetically encoded amino acids, amino acid residues in the peptides may be substituted with naturally occurring non-encoded amino acids and synthetic amino acids. Certain commonly encountered amino acids which provide useful substitutions include, but are not limited to, β-alanine and other omega-amino acids, such as 3- aminopropionic acid, 2,3-diaminopropionic acid, 4-aminobutyric acid and the like; α- aminoisobutyric acid; ε-aminohexanoic acid; δ-aminovaleric acid; N-methylglycine or sarcosine; ornithine; citrulline; t-butylalanine; t-butylglycine; N-methylisoleucine; phenylglycine; cyclohexylalanine; norleucine; naphthylalanine; 4-chlorophenylalanine; 2-fluorophenylalanine; 3- fluorophenylalanine; 4-fluorophenylalanine; penicillamine; l,2,3,4-tetrahydroisoquinoline-3- carboxylic acid; β 2-thienylalanine; methionine sulfoxide; homoarginine; N-acetyl lysine; 2,4- diaminobutyric acid; 2,3-diaminobutyric acid; p-aminophenylalanine; N-methyl valine; homocysteine; homophenylalanine; homoserine; hydroxyproline; homoproline; N-methylated amino acids; and peptoids (N-substituted glycines).
While in certain embodiments, the amino acids of the peptides will be substituted with L- amino acids, the substitutions are not limited to L-amino acids. Thus, also encompassed by the present disclosure are modified forms of the peptides, wherein an L-amino acid is replaced with an identical D-amino acid (e.g., L-Arg— >D-Arg) or with a conservatively-substituted D-amino acid (e.g., LArg— >D-Lys), and vice versa.
Additional aspects of the disclosure include analogs, variants, derivatives, and mimetics based on the amino acid sequence of the peptides disclosed herein. Typically, mimetic compounds are synthetic compounds having a three-dimensional structure (of at least part of the mimetic compound) that mimics, for example, the primary, secondary, and/or tertiary structural, and/or electrochemical characteristics of a selected peptide, structural domain, active site, or binding region (e.g., a homotypic or heterotypic binding site, a catalytic active site or domain, a receptor or ligand binding interface or domain, or a structural motif) thereof. The mimetic compound will often share a desired biological activity with a native peptide, as discussed herein (e.g., the ability to interact with lipids). Typically, at least one subject biological activity of the mimetic compound is not substantially reduced in comparison to, and is often the same as or greater than, the activity of the native peptide on which the mimetic was modeled.
A variety of techniques well known to one of skill in the art are available for constructing synthetic peptide mimetics with the same, similar, increased, or reduced biological activity as the corresponding native peptide. Often these analogs, variants, derivatives and mimetics will exhibit one or more desired activities that are distinct or improved from the corresponding native peptide, for example, improved characteristics of solubility, stability, lipid interaction, and/or susceptibility to hydrolysis or proteolysis (see, e.g., Morgan and Gainor, Ann. Rep. Med. Chem. 24:243-252, 1989). In addition, mimetic compounds of the disclosure can have other desired characteristics that enhance their therapeutic application, such as increased cell permeability, greater affinity and/or avidity for a binding partner, and/or prolonged biological half-life. The mimetic compounds of the disclosure can have a backbone that is partially or completely non- peptide, but with side groups identical to the side groups of the amino acid residues that occur in the peptide on which the mimetic compound is modeled. Several types of chemical bonds, for example, ester, thioester, thioamide, retroamide, reduced carbonyl, dimethylene and ketomethylene bonds, are known in the art to be generally useful substitutes for peptide bonds in the construction of protease-resistant mimetic compounds.
In one embodiment, peptides useful within the disclosure are modified to produce synthetic peptide mimetics by replacement of one or more naturally occurring side chains of the 20 genetically encoded amino acids (or D-amino acids) with other side chains, for example with groups such as alkyl, lower alkyl, cyclic 4-, 5-, 6-, to 7-membered alkyl, amide, amide lower alkyl, amide di(lower alkyl), lower alkoxy, hydroxy, carboxy and the lower ester derivatives thereof, and with 4-, 5-, 6-, to 7-membered heterocyclics. For example, proline analogs can be made in which the ring size of the proline residue is changed from a 5-membered ring to a 4-, 6-, or 7-membered ring. Cyclic groups can be saturated or unsaturated, and if unsaturated, can be aromatic or non-aromatic. Heterocyclic groups can contain one or more nitrogen, oxygen, and/or sulphur heteroatoms. Examples of such groups include furazanyl, furyl, imidazolidinyl, imidazolyl, imidazolinyl, isothiazolyl, isoxazolyl, morpholinyl (e.g., morpholino), oxazolyl, piperazinyl (e.g., 1-piperazinyl), piperidyl (e.g., 1-piperidyl, piperidino), pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridyl, pyrinidinyl, pyrrolidinyl (e.g., 1- pyrrolidinyl), pyrrolinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, thiomorpholinyl (e.g., thiomorpholino), and thiazolyl groups. These heterocyclic groups can be substituted or unsubstituted. Where a group is substituted, the substituent can be alkyl, alkoxy, halogen, oxygen, or substituted or unsubstituted phenyl. Peptides, as well as peptide analogs and mimetics, can also be covalently bound to one or more of a variety of nonproteinaceous polymers, for example, polyethylene glycol, polypropylene glycol, or polyoxyalkenes, as described in U.S. Pat. Nos. 4,640,835; 4,496,689; 4,301,144; 4,670,417; 4,791,192; and 4,179,337.
Other peptide analogs and mimetics within the scope of the disclosure include glycosylation variants, and covalent or aggregate conjugates with other chemical moieties. Covalent derivatives can be prepared by linkage of functionalities to groups which are found in amino acid side chains or at the N- or C-termini, by means which are well known in the art. These derivatives can include, without limitation, aliphatic esters or amides of the carboxyl terminus, or of residues containing carboxyl side chains, O-acyl derivatives of hydroxyl group- containing residues, and N-acyl derivatives of the amino terminal amino acid or amino-group containing residues (e.g., lysine or arginine). Acyl groups are selected from the group of alky 1- moieties including C3 to Cl 8 alkyl, thereby forming alkanoyl aroyl species. Also embraced are versions of a native primary amino acid sequence which have other minor modifications, including phosphorylated amino acid residues, for example, phosphotyrosine, phosphoserine, or phosphothreonine, or other moieties, including ribosyl groups or cross-linking reagents.
In the peptides disclosed herein, the linkage between amino acid residues can be a peptide bond or amide linkage (e.g., -C-C(O)NH-). Alternatively, one or more amide linkages is optionally replaced with a linkage other than amide, for example, a substituted amide. Substituted amides generally include, but are not limited to, groups of the formula -C(O)NR-, where R is (Cj-C6) alkyl, substituted (Ci-C6) alkyl, (C1-C6) alkenyl, substituted (C1-C6) alkenyl, (Ci-C6) alkynyl, substituted (C1-C6) alkynyl, (C5-C20) aryl, substituted (C5-C20) aryl, (C6-C26) alkaryl, substituted (C6-C26) alkaryl, 5-20 membered heteroaryl, substituted 5-20 membered heteroaryl, 6-26 membered alkheteroaryl, and substituted 6-26 membered alkheteroaryl. Additionally, one or more amide linkages can be replaced with peptidomimetic or amide mimetic moieties which do not significantly interfere with the structure or activity of the peptides. Suitable amide mimetic moieties are described, for example, in Olson et ah, J. Med. Chem. 36:3039-3049, 1993. The peptides of the present invention may optionally be acetylated at the N-terminus. The peptides of the present invention may optionally have a carboxy terminal amide. In some embodiments, the peptides of the present invention may have both an acetylated N-terminus and a carboxy terminal amide. Methods of acetylating the N-terminus or adding a carboxy terminal amide are well known to one of ordinary skill in the art. E. Labeled Peptides
In another embodiment, a detectable moiety can be linked to any of the compositions disclosed herein, creating a composition/detectable moiety conjugate. The compositions disclosed herein may be labeled using labels and techniques known to one of ordinary skill in the art. Some of these labels are described in the "Handbook of Fluorescent Probes and Research Products", ninth edition, Richard P. Haugland (ed) Molecular Probes, Inc. Eugene, OR), which is incorporated herein in its entirety. Detectable moieties suitable for such use include any composition detectable by spectroscopic, photochemical, biochemical, immunochemical, electrical, optical, magnetic or chemical means. The detectable moieties contemplated for the present disclosure can include, but are not limited to, an immunofluorescent moiety (e.g., fluorescein, rhodamine, Texas red, and the like), a radioactive moiety (e.g., 3H, 32P, 1251, 1311, 35S), an enzyme moiety (e.g., horseradish peroxidase, alkaline phosphatase), a colorimetric moiety (e.g., colloidal gold, biotin, colored glass or plastic, and the like). The detectable moiety can be linked to the composition. Optionally, a linker can be included between the composition and the detectable moiety.
Means of detecting such moieties are well known to those of skill in the art. Thus, for example, radiolabels may be detected using photographic film, gamma counters or scintillation counters. Fluorescent markers may be detected using a photodetector to detect emitted illumination. Enzymatic labels are typically detected by providing the enzyme with a substrate and detecting the reaction product produced by the action of the enzyme on the substrate, and colorimetric labels are detected by simply visualizing the colored label.
IV. Synthesis and Purification of HDL Mimetic Peptides The HDL mimetic peptides peptides or peptide analogs of the invention can be prepared using virtually any technique known to one of ordinary skill in the art for the preparation of peptides. For example, the peptides can be prepared using step-wise solution or solid phase peptide syntheses, or recombinant DNA techniques, or the equivalents thereof
A. Chemical Synthesis
HDL mimetic peptides of the invention containing amino acids having either the D- or L- configuration can be readily synthesized by automated solid phase procedures well known in the art. Suitable syntheses can be performed by utilizing "T-boc" or "F-moc" procedures. Techniques and procedures for solid phase synthesis are described in Solid Phase Peptide Synthesis: A Practical Approach, by E. Atherton and R. C. Sheppard, published by IRL, Oxford University Press, 1989. Alternatively, the peptides may be prepared by way of segment condensation, as described, for example, in Liu et ah, Tetrahedron Lett. 37:933-936, 1996; Baca et al, J. Am. Chem. Soc. 117:1881-1887, 1995; Tarn et al, Int. J. Peptide Protein Res. 45:209- 216, 1995; Schnolzer and Kent, Science 256:221-225, 1992; Liu and Tarn, J Am. Chem. Soc. 116:4149-4153, 1994; Liu and Tarn, Proc. Natl. Acad. ScL USA 91:6584-6588, 1994; and Yamashiro and Li, Int. J. Peptide Protein Res. 31:322-334, 1988). This is particularly the case with glycine containing peptides. Other methods useful for synthesizing the peptides of the disclosure are described in Nakagawa et al., J. Am. Chem. Soc. 107:7087-7092, 1985. Additional exemplary techniques known to those of ordinary skill in the art of peptide and peptide analog synthesis are taught by Bodanszky, M. and Bodanszky, A., The Practice of Peptide Synthesis, Springer Verlag, New York, 1994; and by Jones, J., Amino Acid and Peptide Synthesis, 2nd ed., Oxford University Press, 2002. The Bodanszky and Jones references detail the parameters and techniques for activating and coupling amino acids and amino acid derivatives. Moreover, the references teach how to select, use and remove various useful functional and protecting groups.
Peptides of the disclosure having either the D- or L-configuration can also be readily purchased from commercial suppliers of synthetic peptides. Such suppliers include, for example, Advanced ChemTech (Louisville, KY), Applied Biosystems (Foster City, CA), Anaspec (San Jose, CA), and Cell Essentials (Boston, MA).
B. Recombinant Synthesis
If the peptide is composed entirely of gene-encoded amino acids, or a portion of it is so composed, the peptide or the relevant portion can also be synthesized using conventional recombinant genetic engineering techniques. For recombinant production, a polynucleotide sequence encoding the peptide is inserted into an appropriate expression vehicle, that is, a vector which contains the necessary elements for the transcription and translation of the inserted coding sequence, or in the case of an RNA viral vector, the necessary elements for replication and translation. The expression vehicle is then transfected into a suitable target cell which will express the peptide. Depending on the expression system used, the expressed peptide is then isolated by procedures well-established in the art. Methods for recombinant protein and peptide production are well known in the art (see, e.g., Sambrook et al. (ed.), Molecular Cloning: A Laboratory Manual, 2nd ed., vol. 1-3, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989, Ch. 17 and Ausubel et al. Short Protocols in Molecular Biology, 4th ed., John Wiley & Sons, Inc., 1999).
To increase efficiency of production, the polynucleotide can be designed to encode multiple units of the peptide separated by enzymatic cleavage sites. The resulting polypeptide can be cleaved {e.g., by treatment with the appropriate enzyme) in order to recover the peptide units. This can increase the yield of peptides driven by a single promoter. In one embodiment, a polycistronic polynucleotide can be designed so that a single mRNA is transcribed which encodes multiple peptides, each coding region operatively linked to a cap-independent translation control sequence, for example, an internal ribosome entry site (IRES). When used in appropriate viral expression systems, the translation of each peptide encoded by the mRNA is directed internally in the transcript, for example, by the IRES. Thus, the polycistronic construct directs the transcription of a single, large polycistronic mRNA which, in turn, directs the translation of multiple, individual peptides. This approach eliminates the production and enzymatic processing of polyproteins and can significantly increase yield of peptide driven by a single promoter. A variety of host-expression vector systems may be utilized to express the peptides described herein. These include, but are not limited to, microorganisms such as bacteria transformed with recombinant bacteriophage DNA or plasmid DNA expression vectors containing an appropriate coding sequence; yeast or filamentous fungi transformed with recombinant yeast or fungi expression vectors containing an appropriate coding sequence; insect cell systems infected with recombinant virus expression vectors (e.g., baculovirus) containing an appropriate coding sequence; plant cell systems infected with recombinant virus expression vectors (e.g., cauliflower mosaic virus (CaMV) or tobacco mosaic virus (TMV)) or transformed with recombinant plasmid expression vectors (e.g., Ti plasmid) containing an appropriate coding sequence; or animal cell systems. The expression elements of the expression systems vary in their strength and specificities.
Depending on the host/vector system utilized, any of a number of suitable transcription and translation elements, including constitutive and inducible promoters, can be used in the expression vector. For example, when cloning in bacterial systems, inducible promoters such as pL of bacteriophage lambda, plac, ptrp, ptac (ptrp-lac hybrid promoter) and the like can be used. When cloning in insect cell systems, promoters such as the baculovirus polyhedron promoter can be used. When cloning in plant cell systems, promoters derived from the genome of plant cells (e.g., heat shock promoters, the promoter for the small subunit of RUBISCO, the promoter for the chlorophyll a/b binding protein) or from plant viruses (e.g., the 35S RNA promoter of CaMV, the coat protein promoter of TMV) can be used. When cloning in mammalian cell systems, promoters derived from the genome of mammalian cells (e.g., metallothionein promoter) or from mammalian viruses (e.g., the adenovirus late promoter, the vaccinia virus 7.5 K promoter) can be used.
C. Purification of HDL Mimetic Peptides
The HDL mimetic peptides or peptide analogs of the invention can be purified by many techniques well known in the art, such as reverse phase chromatography, high performance liquid chromatography, ion exchange chromatography, size exclusion chromatography, affinity chromatography, gel electrophoresis, and the like. The actual conditions used to purify a particular peptide or peptide analog will depend, in part, on synthesis strategy and on factors such as net charge, hydrophobicity, hydrophilicity, and the like, and will be apparent to those of ordinary skill in the art.
For affinity chromatography purification, any antibody which specifically binds the peptide or peptide analog may be used. The peptides of the present invention may optionally be acetylated at the N-terminus. The peptides of the present invention may optionally have a carboxy terminal amide. In some embodiments, the peptides of the present invention may have both an acetylated N-terminus and a carboxy terminal amide. Methods of acetylating the N-terminus or adding a carboxy terminal amide are well known to one of ordinary skill in the art.
V. Pharmaceutical Compositions and Uses Thereof
The compositions of the disclosure can be used, alone or in combination, together with a pharmaceutically acceptable carrier, to treat any disorder in animals, especially mammals (e.g., humans), for increasing HDL levels or promoting lipid efflux is beneficial. Such disorders include, but are not limited to, dyslipidemic and vascular disorders.
This method includes administering to the animal or the human a therapeutically effective amount of a pharmaceutical composition of the present invention. In specific, non-limiting examples, the dyslipidemic and vascular disorders include hyperlipidemia (e.g., hypercholesterolemia), hyperlipoproteinemia, hypercholesterolemia, hypertriglyceridemia, HDL deficiency, apoAl deficiency, cardiovascular disease, cerebrovascular disease, coronary artery disease, atherosclerosis, myocardial infarction, stroke, thrombotic stroke, peripheral vascular disease, restenosis (e.g., atherosclerotic plaques), acute coronary syndrome, and reperfusion myocardial injury. The compositions of the disclosure can also be used alone or in combination during the treatment of thrombotic stroke, infarcts secondary to occlusion of a vessel and during thrombolytic treatment of occluded coronary artery disease. The compositions of the disclosure can be used to treat tissue following hypoxia, ischemia and infarction due to impairment of blood supply, and also following hemorrhage following rupture or trauma of a blood vessel. Such tissue includes, without limitation, neural tissue in the central or peripheral nervous system, peripheral vascular tissue, and cardiac muscle.
The present compositions can be used alone or in combination therapy with other lipid- lowering compositions or drugs and/or other anti-inflammatory compositions or drugs used to treat the foregoing conditions. Such therapies include, but are not limited to simultaneous or sequential administration of the drugs involved. For example, in the treatment of hypercholesterolemia or atherosclerosis, the pharmaceutical composition formulations can be administered with any one or more of the cholesterol lowering therapies currently in use, for example, bile-acid resins, niacin, statins, fat uptake inhibitors, and HDL-raising drugs. In another embodiment, the present compositions can be used in conjunction with statins or fibrates to treat hyperlipidemia, hypercholesterolemia and/or cardiovascular disease, such as atherosclerosis. In yet another embodiment, the compositions of the disclosure can be used in combination with an anti-microbial agent and/or an anti-inflammatory agent, such as aspirin. In another embodiment compositions of the disclosure can be used in combination with anti- hypertensive medicines known to one of ordinary skill in the art. It is to be understood that more than one additional therapy may be combined with administration of the compositions of the disclosure.
In yet another embodiment, the present compositions can be used in conjunction with medicines used to treat patients with cerebrovascular and cardiovascular disease resulting in hypoxia, ischemia and infarction due to impairment of blood supply, and also following hemorrhage following rupture or trauma of a blood vessel. Such medicines are commonly known to one of ordinary skill in the art and include without limitation, modulators of excitatory amino acids and modulators of platelet aggregation.
A. Administration of the Compositions of the Present Invention
In exemplary applications, pharmaceutical compositions comprising the compositions of the present invention in an acceptable carrier are administered to an animal or a human suffering from a dyslipidemic or vascular disorder, such as hyperlipidemia, hyperlipoproteinemia, hypercholesterolemia, hypertriglyceridemia, HDL deficiency, apoAl deficiency, coronary artery disease, atherosclerosis, stroke, ischemia, infarction, myocardial infarction, hemorrhage, peripheral vascular disease, restenosis, acute coronary syndrome, or reperfusion myocardial injury, in an amount sufficient to inhibit or treat the dyslipidemic or vascular disorder. Amounts effective for this use will depend upon the severity of the disorder and the general state of the subject's health. A therapeutically effective amount of the compound is that which provides either subjective relief of a symptom(s) or an objectively identifiable improvement as noted by the clinician or other qualified observer.
A pharmaceutical composition can be administered by any means known to one of skill in the art, such as by oral, intramuscular, subcutaneous, intracardiac, intraarterial or intravenous injection, but even nasal or anal administration is contemplated. In one embodiment, administration is by oral administration. In one embodiment, administration is by subcutaneous or intramuscular injection. To extend the time during which the composition is available to inhibit or treat a dyslipidemic or vascular disorder, the composition can be provided as an implant, an oily injection, or as a particulate system. The particulate system can be a vesicle, microparticle, a microcapsule, a microsphere, a nanoparticle, or similar particle (Banga, "Parenteral Controlled Delivery of Therapeutic Peptides and Proteins," in Therapeutic Peptides and Proteins, Technomic Publishing Co., Inc., Lancaster, PA, 1995). The present compositions may also be applied to a medical device for delivery to a specific location. For example, a stent, a surgical tool, catheter, balloon, electrode, suture, or an artificial vessel or transplanted vessel may contain or be coated with the present composition. In one embodiment, vesicles, such as nanoparticles containing the HDL mimetic peptides are embedded into the stent and released when the stent is located within a desired vessel to promote lipid efflux and/or HDL release.
B. Representative Methods of Administration, Formulations and Dosage The compositions of the present invention can be combined with a pharmaceutically acceptable carrier {e.g., a phospholipid or other type of lipid) or vehicle for administration to human or animal subjects. In some embodiments, more than one composition can be combined to form a single preparation. The compositions can be conveniently presented in unit dosage form and prepared using conventional pharmaceutical techniques. Such techniques include the step of bringing into association the active ingredient and the pharmaceutical carrier(s) or excipient(s). In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers. Formulations suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The formulations may be presented in unit- dose or multi-dose containers, for example, sealed ampules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of a sterile liquid carrier, for example, water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets commonly used by one of ordinary skill in the art.
In certain embodiments, unit dosage formulations are those containing a dose or unit, or an appropriate fraction thereof, of the administered ingredient. It should be understood that in addition to the ingredients particularly mentioned above, formulations encompassed herein may include other agents commonly used by one of ordinary skill in the art.
The pharmaceutical compositions provided herein, including those for use in treating dyslipidemic and vascular disorders, may be administered through different routes, such as oral, including buccal and sublingual, rectal, parenteral, aerosol, nasal, intramuscular, intraperitoneal, intravascular, subcutaneous, intradermal, and topical. They may be administered in different forms, including but not limited to solutions, emulsions and suspensions, microspheres, vesicles, particles, microparticles, nanoparticles, and liposomes.
In another embodiment, it may be desirable to administer the pharmaceutical compositions locally to the area in need of treatment. This maybe achieved by, for example, and not by way of limitation, local or regional infusion or perfusion during surgery, direct perfusion into the heart, particularly the left ventricle, direct perfusion into a vessel, such as the aorta, celiac trunk, common hepatic artery, right or left hepatic artery, portal vein or any of the veins leading to the portal vein, an atherosclerotic vessel, topical application (e.g., wound dressing, coated stent), injection, catheter, suppository, or implant (e.g., implants formed from porous, non- porous, or gelatinous materials, including membranes, such as silastic membranes or fibers), and the like. In one embodiment, administration can be by direct injection at the site (or former site) of a tissue that is to be treated, such as the liver, heart or the peripheral vasculature. In another embodiment, the pharmaceutical compositions are delivered in a vesicle, in particular liposomes (see, e.g., Langer, Science 249:1527-1533, 1990; Treat et al, in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, N. Y., pp. 353-365, 1989) or nanoparticles. Combinations of administration methods may also be employed such as oral, systemic and/or local infusion of a pharmaceutical composition of the present invention, before, after or during placement of a stent coated with a pharmaceutical composition of the present invention. Stents containing the compositions of the present invention may be used at any location where treatment is desired, e.g. but not limited to a coronary artery or a carotid artery. Any vessel in the cardiovascular system that needs treatment may be treated.
In yet another embodiment, the pharmaceutical compositions can be delivered in a controlled release system. In one embodiment, a pump can be used (see, e.g., Longer Science 249:1527-1533, 1990; Sefton Crit. Rev. Biomed. Eng. 14:201-240, 1987; Buchwald et al, Surgery 88:507-516, 1980; Saudek et al, N. Engl. J. Med. 321:574-579, 1989). In another embodiment, polymeric materials can be used (see, e.g., Ranger et al, Macromol ScL Rev. Macromol. Chem. 23:61-64, 1983; Levy et al, Science 228:190-192, 1985; During et al, Ann. Neurol. 25:351-356, 1989; and Howard et al, J. Neurosurg. 71:105-112, 1989). Other controlled release systems, such as those discussed in the review by Langer (Science 249:1527-1533, 1990), can also be used.
The amount of the pharmaceutical compositions that will be effective depends on the nature of the disorder or condition to be treated, as well as the stage of the disorder or condition. Effective amounts can be determined by standard clinical techniques. The precise dose to be employed in the formulation will also depend on the route of administration, and should be decided according to the judgment of the health care practitioner and each subject's circumstances. All dosage forms and therapeutically effective amounts of the compositions of the present invention may be employed. An example of such a dosage range is 0.1 to 200 mg/kg body weight in single or divided doses. Another example of a dosage range is 1.0 to 100 mg/kg body weight in single or divided doses.
The specific dose level and frequency of dosage for any particular subject may be varied and will depend upon a variety of factors, including the activity of the specific compound, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, and severity of the condition of the subject undergoing therapy.
The pharmaceutical compositions of the present disclosure can be administered at about the same dose throughout a treatment period, in an escalating dose regimen, or in a loading-dose regime (e.g., in which the loading dose is about two to five times the maintenance dose). In some embodiments, the dose is varied during the course of a treatment based on the condition of the subject being treated, the severity of the disease or condition, the apparent response to the therapy, and/or other factors as judged by one of ordinary skill in the art. The volume of administration will vary depending on the route of administration. By way of example, oral administration may employ any orally acceptable dosage form commonly known to one of ordinary skill in the art (pill, lozenge, flash melt formulation, time release capsules, liquid, syrup, emulsion, etc.) and any therapeutically effective amount of the compositions of the present invention. By way of example, intramuscular injections may range from about 0.1 ml to about 1.0 ml. Those of ordinary skill in the art will know appropriate volumes for different routes of administration.
The following examples will serve to further illustrate the present invention without, at the same time, however, constituting any limitation thereof. On the contrary, it is to be clearly understood that resort may be had to various embodiments, modifications, and equivalents thereof which, after reading the description herein, may suggest themselves to those skilled in the art without departing from the spirit of the invention.
The subject matter of the present disclosure is further illustrated by the following non- limiting Examples.
Example 1: Administration of the compositions of the present invention to increase lipid efflux and HDL in humans
An individual with low plasma levels of apoAl and HDL, relatively high levels of LDL, and high cholesterol is treated with the pharmaceutical compositions of the present invention through a daily oral administration. After a week, plasma levels of HDL are increased and total cholesterol is decreased.
Example 2: Intravenous administration of the compositions of the present invention to treat atherosclerosis in humans
Individuals with acute coronary syndrome and documented atherosclerosis have a cardiac catherization with intravascular ultrasound (IVUS) to document coronary atherosclerosis of 20 to 50% obstruction in the target artery . Each individual is on stable hypolipidemic drug therapy and receives an acceptable dose of a pharmaceutical composition of the present invention iv weekly for a period of 5 to 8 weeks. A repeat IVUS measurement is made at the end of the treatment period to assess the effect of the pharmaceutical composition infusion on coronary atherosclerosis in the target vessel. Plaque is reduced in the atherosclerotic coronary artery following the treatment, demonstrating efficacy of the pharmaceutical compositions of the present invention to treat atherosclerosis. Example 3: Oral administration of the compositions of the present invention to treat atherosclerosis in humans
Individuals with acute coronary syndrome and documented atherosclerosis have a cardiac catheterization with intravascular ultrasound (IVUS) to document coronary atherosclerosis of 20 to 50% obstruction in the target artery. Each individual is on stable hypolipidemic drug therapy and receives an acceptable dose of a pharmaceutical composition of the present invention by mouth weekly for a period of 5 to 8 weeks. A repeat IVUS measurement is made at the end of the treatment period to assess the effect of the oral administration of the pharmaceutical composition on coronary atherosclerosis in the target vessel. Plaque is reduced in the atherosclerotic coronary artery following the treatment demonstrating efficacy of the pharmaceutical compositions of the present invention to treat atherosclerosis.
Example 4: Administration of the pharmaceutical compositions of the present invention to prevent atherosclerosis in humans Individuals with documented risk factors for atherosclerosis and having high plasma cholesterol levels have a ultrasound analysis of the coronary (IVUS), carotid (IMT), or popliteal arteries to establish a baseline measurement. A portion of these individuals are daily administered individual pharmaceutical compositions of the present invention at a dose of 2 mg/kg to 50 mg/kg intravenously (iv) or intramuscularly (im) 1 to 3 times per week over a period of approximately one to six months. Alternatively, each individual receives a suitable oral dose of such a pharmaceutical composition daily for approximately one to six months.
The other individuals receive a control composition. A new ultrasound analysis at the end of the treatment period indicates higher levels of plaque in the vessels of individuals receiving the control composition, relative to individuals receiving a pharmaceutical composition of the present invention. This example indicates that the individual compositions of the present invention are effective in preventing or reducing atherosclerosis in individuals at risk for developing atherosclerosis and in reducing plaque accumulation in coronary, carotid, or popliteal arteries. Example 5: Administration of the compositions of the present invention on stents and/or orally, to reduce inflammation and restenosis
Individuals with acute coronary syndrome and having plaque in coronary vessels that requires a stent to reduce the obstruction receive an IVUS procedure to document the coronary obstruction. These individuals are divided into three groups. One group receives a stent coated with a peptide of the present invention. A second group receives an oral administration of the composition of the present invention at a dose of 2 mg/kg to 50 mg/kg, 2 to 3 times per week over a period of approximately 6 to 10 weeks. A third group receives a stent coated with a peptide of the present invention and an oral administration of the composition of a peptide of the present invention at a dose of 2 mg/kg to 50 mg/kg, 2 to 3 times per week over a period of approximately 6 to 10 weeks.
All individuals receive a second IVUS procedure at the end of 6 or 10 weeks. The results demonstrate that individuals receiving either a coated stent, a coated stent plus oral administration, or oral administration alone, all display reduced inflammation and restenosis when compared to their condition at the time of the first IVUS procedure.
Example 6: Administration of the pharmaceutical compositions of the present invention to prevent myocardial infarction or stroke in humans
Individuals with documented risk factors for myocardial infarction or stroke (including those individuals who have already had a myocardial infarction or stroke) are divided into five groups. A first group receives a stent coated with an encapsulated peptide of the present invention. A second group receives an orally-administered encapsulated peptide of the present invention at a dose of about 2 mg/kg to 50 mg/kg, two to three times per week over a period of about six months to about thirty-six months. A third group receives the coated stent plus the orally administered encapsulated peptide. A fourth group receives an uncoated stent. A fifth group receives an orally administered control composition (e.g., a placebo).
After about six months to thirty-six months, there are fewer occurrences of myocardial infarction or stroke in the first, second, and third groups of individuals (who had been treated with the peptide-coated stent, oral peptide composition, or both), compared to the fourth and fifth groups of individuals (who had been treated with the uncoated stent or the oral control composition). This result demonstrates that treatment with an encapsulated peptide of the present invention: 1) delays or prevents onset of an initial myocardial infarction or stroke and/or 2) delays or prevents recurrence of myocardial infarction or stroke. All patents, publications and abstracts cited above are incorporated herein by reference in their entirety. It should be understood that the foregoing relates only to preferred embodiments of the present invention and that numerous modifications or alterations may be made therein without departing from the spirit and the scope of the present invention as defined in the following claims. It will be apparent that the precise details of the constructs, compositions, and methods described herein may be varied or modified without departing from the spirit of the described invention. We claim all such modifications and variations that fall within the scope and spirit of the claims below.

Claims

I. A composition comprising a high density lipoprotein (HDL) mimetic peptide encapsulated within a vesicle.
2. The composition of claim 1, wherein the vesicle comprises lipid.
3. The composition of claim 1 , wherein the vesicle comprises a nanoparticle.
4. The compositions of claim 1, wherein the HDL mimetic peptide comprises at least one of SEQ ID NOs: 1 through 400, or an amino acid substitution of at least one of SEQ ID
NOs: 1 through 400.
5. The composition of claim 4, wherein the HDL mimetic peptide comprises at least one of SEQ ID NOs: 1-17, 21-37, 41-57, 61-77, 81-148, 161-228, 241-308, and 321-388, or an amino acid substitution of at least one of SEQ ID NOs: 1-17, 21-37, 41-57, 61-77, 81- 148, 161-228, 241-308, and 321-388.
6. The composition of claim 4, wherein the HDL mimetic peptide comprises at least one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ED NO:
I 1, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, and SEQ ID NO: 20.
7. The composition of claim 6, wherein the HDL mimetic peptide comprises at least one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ DD NO: 5, SEQ
DD NO: 6, SEQ ID NO: 7, SEQ DD NO: 8, SEQ ID NO: 9, SEQ DD NO: 10, SEQ DD NO: 11, SEQ DD NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ DD NO: 16, and SEQ DD NO: 17.
8. The composition of claim 4, wherein the HDL mimetic peptide comprises at least one of SEQ DD NO: 61, SEQ DD NO: 62, SEQ DD NO: 63, SEQ DD NO: 64, SEQ DD NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, and SEQ ID NO: 80.
9. The composition of claim 8, wherein the HDL mimetic peptide comprises at least one of SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ED NO: 68, SEQ ID NO: 69, SEQ ED NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, and SEQ ID NO: 77.
10. The composition of claim 4, wherein the HDL mimetic peptide comprises at least one of SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, SEQ ED NO: 244, SEQ ED NO: 245, SEQ ED NO: 246, SEQ ED NO: 247, SEQ ED NO: 248, SEQ ED NO: 249, SEQ ID NO: 250, SEQ ED NO: 251, SEQ ED NO: 252, SEQ ED NO: 253, SEQ ED NO: 254, SEQ ED NO: 255, SEQ ED NO: 256, SEQ ED NO: 257, SEQ ID NO: 309, SEQ ED NO: 310, and SEQ ED NO: 311.
11. The composition of claim 10, wherein the HDL mimetic peptide comprises at least one of SEQ ED NO: 241, SEQ ED NO: 242, SEQ ID NO: 243, SEQ ED NO: 244, SEQ ID NO: 245, SEQ ED NO: 246, SEQ ED NO: 247, SEQ ED NO: 248, SEQ ED NO: 249, SEQ ED NO: 250, SEQ ED NO: 251, SEQ LD NO: 252, SEQ ED NO: 253, SEQ ED NO: 254, SEQ ED NO: 255, SEQ ID NO: 256, and SEQ ID NO: 257.
12. A pharmaceutical composition comprising the composition of any of the preceding claims and a pharmaceutically acceptable carrier.
13. Use of the composition of any of claims 1 through 11 in the manufacture of a medicament for increasing lipid efflux or increasing HDL levels in an animal or human recipient of the medicament.
14. Use of the composition as in claim 13, wherein the medicament is manufactured for oral administration.
15. Use of the composition as in claim 13, wherein the medicament is manufactured for coating a vascular stent.
16. Use of the composition of any one of claims 1 through 11 in the manufacture of a medicament for treating a dyslipidemic or vascular disorder in an animal or human recipient of the medicament.
17. Use of the composition as in claim 16, wherein the medicament is manufactured for oral administration.
18. Use of the composition as in claim 16, wherein the medicament is manufactured for coating a vascular stent.
19. A method for increasing lipid efflux or increasing HDL levels in an animal or a human, comprising administering an amount of the composition of claim 12 to the animal or the human, wherein the amount is effective to increase lipid efflux or to increase HDL levels in the animal or the human, thereby increasing lipid efflux or increasing HDL levels in the animal or the human.
20. The method of claim 19, wherein the composition is orally administered to the animal or the human.
21. The method of claim 19, wherein the composition is administered as a coating on a vascular stent inserted into a blood vessel of the animal or the human.
22. A method for treating or ameliorating a dyslipidemic or vascular disorder in an animal or a human comprising administering an amount of the composition of claim 12 to the animal or the human, wherein the amount is effective to treat or ameliorate the dyslipidemic or vascular disorder in the animal or the human, thereby treating or ameliorating the dyslipidemic or vascular disorder in the animal or the human.
23. The method of claim 22, wherein the composition is orally administered to the animal or the human.
24. The method of claim 22, wherein the composition is administered as a coating on a vascular stent inserted into a blood vessel of the animal or the human.
25. The method of claim 22, wherein the dyslipidemic disorder or the vascular disorder comprises hyperlipidemia, hyperlipoproteinemia, hypercholesterolemia, hypertriglyceridemia, HDL deficiency, apoA-I deficiency, coronary artery disease, atherosclerosis, thrombotic stroke, peripheral vascular disease, restenosis, acute coronary syndrome, post-perfusion myocardial injury, stroke, ischemia, myocardial infarction, or a combination thereof.
26. The method of claim 22, further comprising administering an additional lipid- lowering composition, an HDL-raising composition, or an inhibitor of fat uptake to the animal or the human.
27. The method of claim 22, further comprising administering an additional antiinflammatory composition to the animal or the human.
28. A vascular stent coated with the composition of any one of claims 1 through 11.
29. A method for increasing lipid efflux or increasing HDL levels in an animal or a human, comprising inserting the vascular stent of claim 28 into a blood vessel of the animal or the human, wherein insertion of the vascular stent is effective to increase lipid efflux or to increase HDL levels in the animal or the human, thereby increasing lipid efflux or increasing HDL levels in the animal or the human.
30. A method for treating or ameliorating a dyslipidemic or vascular disorder in an animal or a human, comprising inserting the stent of claim 28 into a vessel of the animal or the human, wherein insertion of the stent is effective to treat or ameliorate the dyslipidemic or vascular disorder in the animal or the human, thereby treating or ameliorating a dyslipidemic or vascular disorder in the animal or the human.
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US11/764,619 2007-06-18
US11/861,758 2007-09-26
US11/861,758 US20080138284A1 (en) 2006-09-26 2007-09-26 Novel Peptides That Promote Lipid Efflux
US2396308P 2008-01-28 2008-01-28
US61/023,963 2008-01-28
US12/021,629 US20080227686A1 (en) 2006-06-16 2008-01-29 Novel Peptides that Promote Lipid Efflux
US12/021,629 2008-01-29
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