EP1610811A2 - Prevention et traitement d'arythmies cardiaques - Google Patents

Prevention et traitement d'arythmies cardiaques

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Publication number
EP1610811A2
EP1610811A2 EP03813770A EP03813770A EP1610811A2 EP 1610811 A2 EP1610811 A2 EP 1610811A2 EP 03813770 A EP03813770 A EP 03813770A EP 03813770 A EP03813770 A EP 03813770A EP 1610811 A2 EP1610811 A2 EP 1610811A2
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EP
European Patent Office
Prior art keywords
xaa
ala
gly
ser
glp
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Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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EP03813770A
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German (de)
English (en)
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EP1610811A4 (fr
Inventor
David R. Hathaway
Alain D. Baron
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Amylin Pharmaceuticals LLC
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Amylin Pharmaceuticals LLC
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Publication of EP1610811A2 publication Critical patent/EP1610811A2/fr
Publication of EP1610811A4 publication Critical patent/EP1610811A4/fr
Withdrawn legal-status Critical Current

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    • 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
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/26—Glucagons
    • 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
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/2278—Vasoactive intestinal peptide [VIP]; Related peptides (e.g. Exendin)
    • 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
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
    • A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin
    • 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
    • A61P9/06—Antiarrhythmics
    • 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

Definitions

  • This invention relates to compositions and methods for preventing cardiac arrhythmias using a compound that binds to a receptor for a glucagon-like peptide-1, an incretin, a glucagon- like ⁇ eptide-1 (GLP-1), an exendin, or an agonist, an analog (preferably an agonist analog), a derivative, or a variant of any of aforementioned compounds and fragments thereof.
  • GLP-1 glucagon-like peptide-1
  • exendin or an agonist, an analog (preferably an agonist analog), a derivative, or a variant of any of aforementioned compounds and fragments thereof.
  • Cardiac arrhythmias and ischemic heart disease afflict an estimated 20 million Americans, and possibly ten times as many people worldwide. If left undetected and untreated, they often result in heart attacks and deaths.
  • An arrhythmia is an irregular heartbeat.
  • the heart beats on its own due to its natural pacemaker, a small cluster of specialized cells called the sinoatrial node (S-A node).
  • S-A node is located in the right atrium and produces electrical signals at regular intervals that are sent through a pathway in the heart muscle.
  • the S-A node signals follow a natural electrical pathway that helps the heart beat efficiently.
  • An electrical impulse travels from the S-A node through the atrioventricular node (A-V node), a second cluster of cells located near the center of the heart. The A-V node then sends the signals out to the walls of the ventricles.
  • A-V node atrioventricular node
  • the two ventricles contract a fraction of a second after they have been filled with blood from an atrial contraction. This timing sequence is called atrio-ventricular synchrony.
  • An arrhythmia can occur when: (1) the S-A node develops an abnormal rate or rhythm; (2) the normal electrical pathway is interrupted, or (3) another part of the heart tries to take over as the pacemaker. Though there are several types of arrhythmias, they all have the commonality of preventing the heart from pumping blood efficiently. Fast, abnormal heart rhythms, usually over 100 beats per minute, are called tachyarrhythmias.
  • ventricular tachycardia a a ⁇ hythmia known as ventricular tachycardia (NT).
  • NT ventricular tachycardia
  • the brain and body may not receive enough blood and oxygen, causing fainting spells, blackouts, temporary blind spots or dizziness.
  • the patient may become unconscious and in extreme cases the heart may stop (cardiac arrest).
  • the most common cause of arrhythmias is heart disease, particularly coronary artery disease, abnormal heart valve function, and heart failure.
  • VT is a frequent precursor to another type of arrhythmia, ventricular fibrillation (VF).
  • VF ventricular fibrillation
  • VF the heart beats much faster than normal, sometimes over 300 beats a minute.
  • the ventricles “quiver” during VF and do not carry out coordinated contractions. Because little blood is pumped from the heart, VF is a form of cardiac arrest and is fatal unless treated immediately.
  • Ventricular tachycardia and fibrillation occur commonly in the setting of ischemic heart disease and congestive heart failure (CHF).
  • CHF congestive heart failure
  • ventricular arrhythmias may develop secondarily to ischemia or reperfusion.
  • Reperfusion occurs subsequent to therapies that reestablish flow in an artery that is obstructed by a blood clot, i.e. thrombolytic agents or following an intervention, such as angioplasty, coronary bypass grafting or placement of an intracoronary stent.
  • a major problem in congestive heart failure is stress hyperglycemia and insulin resistance.
  • stress hyperglycemia and insulin resistance As a result of the combination of high circulating levels of free fatty acids and reduced glucose uptake, there is a shift toward fatty acid oxidation, depletion of Krebs cycle intermediates and diminished glucose oxidation. These changes ultimately lead to reduced levels of CrP and loss of energy reserve.
  • Pacemakers are electronic devices that act in place of the heart's own pacemaker and are programmed to imitate the normal conduction sequence of the heart. Usually they are implanted surgically beneath the skin of the chest and have wires running to the heart. There are several disadvantages associated with the use of pacemakers, including the need to replace the units every 8-10 years and their potential to be interfered with by certain types of equipment, such as magnetic resonance imaging machines (MRIs).
  • MRIs magnetic resonance imaging machines
  • Therapy for arrhythmias can also include devices that deliver a shock to the heart to stop an abnormal rhythm and restore a normal one.
  • Using an electric shock for this purpose is called cardioversion, electroversion, or defibrillation.
  • a large machine that delivers a shock (a defibrillator) is used by a team of doctors and nurses to stop a life-threatening arrhythmia.
  • a defibrillator about the size of a pack of cards can be implanted surgically in the patient.
  • These small devices which automatically sense life-threatening arrhythmias and deliver a shock, are used in people who would otherwise die when their heart suddenly stops. Because these defibrillators do not prevent arrhythmias, the patient usually must also take drugs as well.
  • compositions of the invention include a compound that binds to a receptor for a glucagon-like peptide-1, an incretin, a glucagon-like peptide-1 (GLP-1), an exendin, or an agonist, an analog (preferably an agonist analog), a derivative, or a variant of any of them, as well as biologically fragments thereof.
  • GLP-1 glucagon-like peptide-1
  • exendin or an agonist, an analog (preferably an agonist analog), a derivative, or a variant of any of them, as well as biologically fragments thereof.
  • compositions of the invention including GLP-1 and exendins
  • GLP-1 has been found to reduce cardiac injury and enhance recovery in patients with these disorders.
  • Incretins including GLP-1, are glucose-dependent insulinotropic hormones.
  • GLP-1 and exendin effectively enhance peripheral glucose uptake without inducing dangerous hypoglycemia. They also strongly suppress glucagon secretion, independent of its insulinotropic action, and thereby powerfully reduce plasma free fatty acid (FFA) levels substantially more than can be accomplished with insulin. High FFA levels have been implicated as a major toxic mechanism during myocardial ischemia.
  • FFA plasma free fatty acid
  • compositions and methods for preventing and treating cardiac arrhythmias Accordingly, it is a primary objective of the present invention to provide compositions and methods for preventing and treating cardiac arrhythmias.
  • the compounds of the invention may be administered by any conventional means, including subcutaneously, intravenously, orally, transmucosally, intraperitoneally, or other means known in the art.
  • the compositions are particularly useful in treating arrhythmias resulting from ischemic heart disease and congestive heart failure.
  • the invention contemplates a method for preventing and treating arrhythmias comprising administering to an individual an effective amount of a composition which includes a compound which binds to a receptor for glucagon-like peptide-1, an incretin, a glucagon-like peptide-1 (GLP-1), an exendin, or an agonist, an analog (preferably an agonist analog), a derivative, or a variant of any of aforementioned compounds, and biologically active fragments thereof.
  • a composition which includes a compound which binds to a receptor for glucagon-like peptide-1, an incretin, a glucagon-like peptide-1 (GLP-1), an exendin, or an agonist, an analog (preferably an agonist analog), a derivative, or a variant of any of aforementioned compounds, and biologically active fragments thereof.
  • methods of the invention include administering compositions of the invention at a dose from about 0.1 pmol/kg/min. up to about 10 pmol/kg/min. Other dose ranges may be from about 0.01 pmol/kg to 20 nmol/kg. Further contemplated are a single or multiple mjection(s) in a dose from about 0.005 nmol/kg to 20 nmol/kg.
  • methods of the invention include a concurrent administration with any one or more of a glucose, a potassium, a free radical scavenger or an anti-oxidant.
  • compositions of the invention are administered within four hours of an ischemic event and may be continued following the ischemic event.
  • the composition may be administered concurrently or as soon as possible following therapies that reestablish flow in an artery that has been obstructed, such as angioplasty, coronary bypass grafting, and placement of an intracoronary stent.
  • compositions of the invention may be administered to treat ventricular arrhythmias.
  • the ventricular arrhythmia may be caused by a condition selected from the group consisting of cardiac ischemia, cardiac ischemia-reperfusion, and congestive heart failure.
  • methods of the invention includemetabolic intervention with a composition that includes a compound which binds to a receptor for glucagon-like peptide-1, an incretin, a glucagon-like peptide-1 (GLP-1), an exendin, or an agonist, an analog (preferably an agonist analog), a derivative, or a variant of any of aforementioned compounds, and fragments thereof to prevent or treat cardiac arrhythmias, said method comprising administering to an individual in need of such treatment an effective amount of a composition which includes a compound which binds to a receptor for glucagon-like peptide-1, an incretin, a glucagon-like peptide-1 (GLP-1), an exendin, or an agonist, an analog (preferably an agonist analog), a derivative, or a variant of any of aforementioned compounds, and biologically active fragments thereof.
  • the present invention relates to the development of compositions for the prevention and treatment of cardiac arrhythmias using an incretin, a glucagon-like peptide-1 (GLP-1), an exendin, a compound that binds to a receptor for glucagon-like peptide-1, or an agonist, an analog (preferably an agonist analog), a derivative, or a variant of any of the aforementioned compounds and biologically active fragments thereof.
  • GLP-1 glucagon-like peptide-1
  • exendin a compound that binds to a receptor for glucagon-like peptide-1
  • an agonist an analog (preferably an agonist analog)
  • a derivative preferably an agonist analog
  • Cardiac arrhythmias can develop due to a variety of factors. For instance, arrhythmias may develop secondary to ischemia or reperfusion. Heart muscle is largely dependent on uninterrupted blood flow, which guarantees delivery of oxygen and substrates to cells while washing out harmful metabolic products. Ischemia, e.g. resulting from decrease or cessation of myocardial blood flow, leads to rapid changes in myocardial metabolism. The degree of these changes is highly dependent upon the severity of the ischemia. For anatomical and physiological reasons, contractile myocytes in endocardium are the most vulnerable cells. Ischemia is a dynamic process. With rapid reperfusion, full recovery of myocardial metabolism occurs; but continuation of ischemia leads to total tissue necrosis in a few hours. Reperfusion, although generally considered beneficial, can cause tissue injury by several mechanisms, including oxidative stress, and thus affect the final recovery of the contractibility.
  • systolic and mitochondrial Ca 2+ levels increase during ischemia and reperfusion.
  • An increase in cystolic Ca + activates a number of channels, carriers, and enzymes and modulates others, which results in delayed afterdepolarizations and arrhythmias.
  • amphiphiles and fatty acids accumulate in the plasma membrane, the gap junction, and the intracellular membranes of the SR and the mitochondria.
  • Amphiphiles and fatty acids may interact directly with channel proteins, with the phospholipids surrounding the channel proteins, or changing the membrane fluidity.
  • Amphiphiles increase inward current at the resting potential with simultaneous reduction of outward current through K + channels.
  • Fatty acids activate outward currents and stimulate the K + /Ca 2+ exchanger. The simultaneous activation of inward and outward currents favors K loss and Ca overload, creating conditions that generate arrhythmias. (Cameliet, 1999).
  • reperfusion injury may manifest clinically as reperfusion arrhythmias.
  • Early reperfusion is an absolute prerequisite for the survival of ischemic tissue. Although ultimately necessary for recovery, reperfusion is often considered a double-edged sword, and can actually lead to worsening of tissue injury by various mechanisms.
  • reperfusion is associated with Ca 2+ overload through activation of the K + /Ca 2+ exchanger, thereby creating conditions favorable to cardiac arrhythmias.
  • Complications associated with congestive heart failure include stress hyperglycemia and insulin resistance.
  • stress hyperglycemia As a result of the combination of high circulating levels of free fatty acids and reduced glucose uptake, there is a shift toward fatty acid oxidation.
  • these fatty acids can activate outward currents through K + channels, and stimulate the K + /Ca 2+ exchanger.
  • the simultaneous activation of inward and outward currents favors K loss and Ca 2+ overload, thereby creating conditions favorable to the generation of arrhythmias.
  • GLP-1 and exendin are glucose-dependent insulinotropic peptides that effectively enhance peripheral glucose uptake without inducing dangerous hypoglycemia. Further, they strongly suppress glucagon secretion, independent of their insulinotropic action, and thereby powerfully reduce plasma free fatty acid (FFA) levels substantially more potently than can be accomplished with insulin (i.e., greater FFA suppression at equivalent prevailing insulin concentrations that are submaximally effective).
  • FFA plasma free fatty acid
  • GLP- 1 exendins, and other compositions of the invention can be effective in the prevention and treatment of cardiac arrhythmias. It has now been found that the dual capacity of GLP-1 to powerfully stimulate insulin release and reduce insulin resistance provides this molecule with the unique ability to prevent and treat cardiac arrhythmias by enhancing glucose uptake and metabolism, at the expense of reduced FAA metabolism, into cardiac muscle.
  • incretins GLP-1 , exendins, compounds that bind a GLP-1 receptor, and agonists, analogs, derivatives, and variants thereof, as well as their active fragments can be especially effective in preventing and treating arrhythmias in patients with cardiac ischemia, cardiac ischemia-reperfusion, and/or congestive heart failure.
  • Treatment with GLP-1 and other compositions of the invention may enhance glycolysis in patients and shift the balance from fatty acid towards glucose oxidation. These effects prevent loss of potassium and calcium overload, and reduce the risk of cardiac arrhythmias.
  • compositions of the invention may also stimulate the secretion of endogenous insulin and therefore can be used to achieve all of the beneficial actions attributed to an insulin infusion in the metabolic treatment of arrhythmias.
  • high-dose GIK infusions typically contain 25-33% glucose and 50-100 U insulin/L, the requirement for introduction of hyperglycemia per se to achieve therapeutic efficacy, versus only providing a metabolic milieu for the safe administration of high doses of insulin, is unclear. It is likely that adequate blood glucose levels are required to enable substrate delivery, but this does not necessarily imply a need for hyperglycemia and should not detract from the fact that insulin exerts important effects other than glucose uptake.
  • a therapeutic infusion of a composition of the invention may require a modest (e.g., 5%) glucose administration in order to maintain blood glucose at slightly above physiological levels in order to trigger insulin release.
  • Glucose is not required as a safety measure, since blood levels of ⁇ 3.5 mM abrogate the insulin-stimulating activity of GLP-1 and exendin, thereby completely protecting against the dangers of hypoglycemia.
  • Insulin resistance has been recognized increasingly as a major pathogenic factor for multiple systemic diseases, and not only in individuals having Type-2 diabetes. Although many patients with Type-2 diabetes manifest insulin resistance, many individuals with IR do not have diabetes. An important recent insight has been the realization that IR is an independent risk factor for the development and severity of cardiovascular diseases, including ischemia- reperfusion injury and left ventricle dysfunction. IR is strongly associated with severe heart disease, both acutely and chronically, which leads to the enhanced and potentially damaging use by the heart of fatty acids as a fuel source in preference to glucose.
  • GLP-1, exendin and other compositions of the invention can reverse the use of fatty acids as fuel to glucose, thereby reducing free fatty acids and preventing the development of conditions favorable to the development of cardiac arrhythmias.
  • the administration of GLP-1, exendin and other compositions of the invention may be especially effective in the treatment of ventricular arrhythmias.
  • GLP-1, exendin, and other compositions of the invention should be effective in a majority of patients without requiring concurrent glucose administration. However, a small proportion of subjects may require glucose to elicit an adequate insulin response. In addition, it also may be necessary to administer potassium to correct excess shifts of potassium in the intracellular compartment when glucose is co-administered with compositions of the invention.
  • the methods of the invention can include use of free radical scavengers or anti-oxidants such as glutathione, melatonin, Vitamin E, and superoxide dismutase (SOD). In such combinations, reperfusion damage risk can be lessened even further.
  • free radical scavengers or anti-oxidants such as glutathione, melatonin, Vitamin E, and superoxide dismutase (SOD).
  • SOD superoxide dismutase
  • compositions of the invention include a compound that binds to a receptor for a glucagon like peptide-1, an incretin, a glucagon-like peptide-1 (GLP-1), an exendm, or an agonist, an analog (preferably an agonist analog), a derivative, or a variant of any of the aforementioned compounds, as well as biologically active fragments thereof.
  • An "agonist” includes any compound that mimics at least one of the actions of an incretin, a GLP-1, or an exendin, as described herein.
  • an "analog” includes any peptide whose sequence was derived from that of the base receptor-binding compound, incretin, GLP-1, or exendin, whether or not including insertions, substitutions, extensions, or deletions, preferably having at least 50 or 55% amino acid sequence identity with the base molecule, more preferably having at least 70%, 80%, 90%, or 95% amino acid sequence identity with the base molecule.
  • Such analogs may comprise conservative or non- conservative amino acid substitutions (including non-natural amino acids or as well as D forms), and if it is an "agonist analog,” exhibits at least one characteristic of the base molecule, preferably having a potency better than the base molecule, or within five orders of magnitude of the base molecule, more preferably 4, 3, 2, or 1 order of magnitude when evaluated by art- known measures.
  • a “derivative” includes any base molecule or analog having a chemical modification within, attached or linked to, or associated with the molecule.
  • Such chemical modifications can include internal linkers (e.g., spacing or structure-inducing) or appended molecules, such as molecular weight-enhancing molecules (e.g., polyethylene glycol (PEG)), or tissue targeting molecules.
  • PEG polyethylene glycol
  • Examples of such molecules are known in the art, for example, insulinotropic peptides, including GLP-1 and exendin, modified with a maleimide group are described in U.S. Patent No. 6,593,295, incorporated herein by reference.
  • variants includes any modification to the base molecule, analog or variant not encompassed in the terms “analog” and “derivative,” as would be known to a person of ordinary skill in the art.
  • variants may include proforms or chimeras of a selected molecule.
  • Small molecules are included in the compounds useful in the invention to the extent that they bind to a receptor for GLP-1 or exendm. Not all of the peptide molecules described as incretins, glucagon-like peptide-1 (GLP-1), exendins, or analogs, derivatives, or variants may bind to a receptor for GLP-1 , although they are still useful in the invention by virtue of a pharmacology not dependent on a known GLP-1 receptor.
  • GLP-1 also known as glucagon-like peptide-1 [7-36] amide (also referred to as GLP-1 [7-36]NH 2 ), a product of the proglucagon gene having the amino acid sequence His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly Gin Ala Ala Lys Glu Phe He Ala Tip Leu Val Lys Gly Arg-NH 2 (SEQ ID NO: 1). It is secreted into plasma mainly from the gut and produces a variety of biological effects related to pancreatic and gastrointestinal function.
  • GLP-1 [7-36]NH 2 , "GLP-1,” as used herein, are known (e.g., Orskov, et al., Diabetes. 42:658-61, 1993; D'Alessio, et al., J. Clin. Invest..
  • compositions of the invention include GLP-1 agonist analogs.
  • agonist analog is meant a compound that mimics at least one effect of GLP-1 as described above. This definition of agonist analog could include compounds that bind to a receptor or receptors where GLP- 1 causes the particular effect.
  • Certain GLP-1 analogs with agonist activity are described in Chen et al, U.S. Patent No. 5,512,549, issued April 30, 1996, entitled Glucagon-Like Insulinotropic Peptide Analogs, Compositions and Methods of Use.
  • Other GLP- 1 analogs with agonist activity are described in Johnson et al, U.S. Patent No.
  • the GLP-1 agonist analogs used in the methods of the present invention can be GLP-l(7-34) and GLP-l(7-35), as disclosed in U.S. Pat. No. 5,118,666, herein incorporated by reference, as well as GLP-l(7-37) as disclosed in U.S. Pat. No: 5,120,712, herein incorporated by reference. Also included are GLP-1 analogs having a reduced tendency to aggregate such as those described in WO 01/98331; GLP-1 analogs that have N-terminal truncation, US Patent No. 5,574,008; GLP-1 analogs with attached acyl groups, US Patent No.
  • analogs include those of formula (XI), Ri -X-Glu-Gly-Thr- Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Y-Gly-Gln-Ala-Ala-Lys-Z -Phe-Ile-Ala-Trp-Leu-Val- Lys-Gly-Arg-R 2 (SEQ IP NO:33) or a pharmacuetically accetable salt thereof, wherein: Ri is selected from the group consisting of His, P-histidine, desamino-histidine, 2-amino- histidine, .beta.-hydroxy-histidine, homohistidine, alpha-fluoromethyl-histidine, and alpha- methyl-histidine; X is selected from the group consisting of Met, Asp, Lys, Thr, Leu, Asn, Gin, Phe, Val, and Tyr
  • Y and Z are independently selected from the group consisting of Glu, Gin, Ala, Thr, Ser, and Gly, and;
  • R 2 is selected from the group consisting of NH 2 , and Gly-OH; provided that, if Ri is His, X is Val, Y is Glu, and Z is Glu, then R 2 is NH 2 .
  • V8-GLP-1 and other position 8 analogs can be found in US Patent No. 5,705,483, incorporated by reference.
  • analogs include those of formula (XII), Ri -X-Glu-Gly-Thr- Phe-Tl r-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Y-Gly-Gln-Ala-Ala-Lys-Z -Phe-Ile-Ala-Trp-Leu-Val- Lys-Gly-Arg-R 2 (SEQ IP NO: 34) wherein:
  • Ri is selected from the group consisting of L-histidine, P-histidine, desamino-histidine, 2- amino-histidine, .beta.-hydroxy-histidine, homohistidine, alpha-fluoromethyl-histidine, and alpha-methyl-histidine;
  • X is selected from the group consisting of Ala, Gly, Val, Thr, He, and alpha-methyl- Ala;
  • Y is selected from the group consisting of Glu, Gin, Ala, Thr, Ser, and Gly
  • Z is selected from the group consisting of Glu, Gin, Ala, Thr, Ser, and Gly;
  • R 2 is selected from the group consisting of NH 2 , and Gly-OH; providing that the compound has an isoelectric point in the range from about 6.0 to about 9.0 and further providing that when Ri is His, X is Ala, Y is Glu, and Z is Glu, R 2 must be NH 2 .
  • the GLP-1 agonist analogs are variants or analogs of GLP-1 known in the art, such as, for example, Gin 9 -GLP-l(7-37), P-Gln 9 -GLP-l(7-37), acetyl-Lys 9 -GLP-1(7- 37), Thr 16 -Lys ls -GLP-1 (7-37), and Lys 18 -GLP-1 (7-37).
  • Perivatives of GLP-1 are also contemplated in the present invention and include, for example, acid addition salts, carboxylate salts, lower alkyl esters, and amides (see, e.g., WO91/11457).
  • GLP-1 GLP-1 agonists of the general formula (I):
  • Ri is selected from the group consisting of 4-imidazopropionyl (des-amino- histidyl), 4-imidazoacetyl, or 4-imidazo-alpha, alpha dimethyl-acetyl;
  • R 2 is selected from the group consisting of C 6 -Cio unbranched acyl, or is absent;
  • R 3 is selected from the group consisting of Gly-OH or NH 2 ;
  • Xaa 0 is Lys or Arg.
  • the GLP-1 agonists are naturally-occurring GLP-1 (7-37) that arise from adding various R groups via a peptide bond to the amino terminus of the peptide portion of Formula I (SEQ IP NO:2).
  • further compounds of the invention are made by acylating the epsilon amino group of the Lys34 residue and by making limited amino acid substitutions at position 26 or by altering the carboxy terminus.
  • the present invention provides biologically-active GLP-1 fragments of formula (II):
  • Ri is selected from the group consisting of: a) H 2 N; b) H 2 N-Ser; c) H 2 N-Val-Ser; d) H 2 N-Asp-Val-Ser; e) H 2 N-Ser-Asp-Val-Ser (SEQ IP NO:4); f) H 2 N-Thr-Ser-Asp-Val-Ser (SEQ IP NO:5); g) H 2 N-Phe-Thr-Ser-Asp-Val-Ser (SEQ IP ⁇ O:6); h) H 2 N-Thr-Phe-Thr-Ser-Asp-Val-Ser (SEQ IP NO:7); i) H 2 N-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser (SEQ IP NO: 8); j) H 2 N-Glu-Gly-Thr-Phe-Thr-Ser-AspNal-Ser (SEQ IP NO:3) wherein Ri is selected from
  • Xaa 41 is selected from the group consisting of Lys or Arg; and wherein R 5 is selected from the group consisting of NH 2 , OH, Gly-NH 2 , or Gly-OH.
  • the invention provides modified forms of the GLP- 1(7-34); (7-35); (7-36) or (7-37) human peptide or the C-terminal amidated forms thereof.
  • the native peptides have the amino acid sequence (SEQ ID NO: 11):
  • modified forms contain one or more alterations of the native structure and are of improved ability for therapeutic use. Either the modified forms have greater potency than glucagon to potentiate insulin secretion or enhanced stability in plasma or both.
  • analogs of the invention which show enhanced insulin stimulating properties may have the foregoing sequence, or a C-terminal amide thereof, with at least one modification of SEQ ID NO: 11, selected from the group consisting of:
  • the substituted amino acids may be in the P form, as indicated by a superscript f , e.g., C .
  • the amino acids substituted at position 7 can also be in the N-acylated or N-alkylated forms.
  • the invention is directed to peptides which show enhanced degradation resistance in plasma as compared to GLP-l(7-37) wherein this enhanced resistance to degradation is defined as set forth below.
  • any of the above-mentioned truncated forms of GLP-1 (7-34) to GLP- 1(7-37) or their C-terminal amidated form is modified by
  • analogs of the invention which are resistant to degradation include (N-acyl (1-6C) AA) 7 GLP-l(7-37) and (N-alkyl (1-6C) AA) 7 GLP-l(7-37) wherein when AA is a lysyl residue, one or both nitrogens may be alkylated or acylated.
  • AA symbolizes any amino acid consistent with retention of insulin stimulating activity.
  • the P residue of any acidic or neutral amino acid can be used at position 7 and of any amino acid at position 8, again consistent with insulin stimulating activity.
  • Either or both of position 7 and 8 can be substituted by a P-amino acid; the P-amino acid at position 7 can also be acylated or alkylated as set forth above.
  • These modified forms are applicable not only to GLP- 1(7-37) but also the shorter truncated analogs as set forth above.
  • Other modified GLP- Is, as well as exendins, useful in the practice of the claimed invention can be found in U.S. Patent No. 6,528,486, which is incorporated by reference.
  • GLP-1 analogs may be peptides containing one or more amino acid substitutions, additions, extensions, or deletions, compared with GLP-1 (7-36)amide, exendin-4 or exendin-3.
  • the number of substitutions, deletions, or additions is 30 amino acids or less, 25 amino acids or less, 20 amino acids or less, 15 amino acids or less, 10 amino acids or less, 5 amino acids or less or any integer in between these amounts.
  • the substitutions include one or more conservative substitutions.
  • a "conservative" substitution denotes the replacement of an amino acid residue by another, biologically active similar residue as is well known in the art.
  • conservative substitutions include the substitution of one hydrophobic residue, such as isoleucine, valine, leucine, or methionine for another, or the substitution of one polar residue for another, such as the substitution of arginine for lysine, glutamic for aspartic acids, or glutamine for asparagine, and the like.
  • GLP-1 analogs include the above described peptides which have been chemically derivatized or altered, for example, peptides with non-natural amino acid residues (e.g., taurine, ⁇ - and ⁇ -amino acid residues and D-amino acid residues), C-terminal functional group modifications, such as amides, esters, and C-terminal ketone modifications and N-terminal functional group modifications, such as acylated amines, Schiff bases, or cyclization, as found, for example, in the amino acid pyroglutamic acid.
  • Exendin analogs may have similar modifications.
  • sequence identity refers to a comparison made between two molecules using standard algorithms well known in the art.
  • the preferred algorithm for calculating sequence identity for the present invention is the Smith- Waterman algorithm, for example, SEQ IP NO: 1 [i.e., GLP- 1(1-37)], SEQ IP NO: 12 or 14 [exendin-3 and 4, respectively] can be used as the reference sequences to define the percentage identity of homology over their length.
  • IP NO: 1 except for 18 amino acid substitutions and an insertion of 3 amino acids, would have a percent identity given by:
  • GLP-1 receptors are cell-surface proteins found, for example, on insulin-producing pancreatic ⁇ -cells; the GLP-l(7-36) receptor has been characterised in the art. . Additional receptors at which GLP-1 and exendins act are also thought to exist, and may mediate effects by which the instant invention is operative. Methods of determining whether a chemical or peptide binds to or activates a particular GLP-1 receptor are known to the skilled artisan. For example, U.S. Patent Nos. 6,051,689, 5,846,747, and 5,670,360 describe GLP-1 receptors, as well as methods for using them. The contents of the patents are incorporated by reference.
  • GLP-1 biological activity can be determined by in vitro and in vivo animal models and human studies, as is well known to the skilled artisan.
  • GLP-1 biological activity can be determined by standard methods, in general, by receptor binding activity screening procedures, which involve providing appropriate cells that express the GLP-1 receptor on their surface, for example, insulinoma cell lines such as RTNmSF cells or INS-1 cells. See Mojsov, Int. J. Peptide Protein Res. 40; 333 (1992) and EP 0708179 A2. Cells that are engineered to express a GLP-1 receptor also can be used.
  • cAMP activity or glucose dependent insulin production can also be measured.
  • a polynucleotide encoding a GLP-1 receptor is employed to tiansfect cells so that they express the GLP-1 receptor protein.
  • these methods may be employed for screening for a receptor agonist by contacting such cells with compounds to be screened and determining whether such compounds generate a signal (i.e., activate the receptor).
  • Other screening techniques include the use of cells that express the GLP-1 receptor, for example, transfected CHO cells, in a system to measure extracellular pH or ionic changes caused by receptor activation.
  • potential agonists may be contacted with a cell that expresses the GLP-1 protein receptor and a second messenger response (e.g., signal transduction or ionic or pH changes), may be measured to determine whether the potential agonist is effective.
  • Polyclonal and monoclonal antibodies can be utilized to detect, purify, and identify GLP-
  • Antibodies such as ABGA1178 detect intact GLP-l(l-37) or N-terminally-truncated GLP-1 (7-37) or GLP-1 (7-36)amide. Other antibodies detect the end of the C-terminus of the precursor molecule, a procedure that allows one — by subtraction — to calculate the amount of biologically active, truncated peptide (i.e., GLP-1 (7-37)amide). Orskov et al, Diabetes 42; 658 (1993); Orskov et al, J. Clin. Invest. 1991, 87; 415 (1991).
  • GLP-1 its agonists, analogs, derivatives, variants, and biologically active fragments, that are peptides can be made by solid-state chemical peptide synthesis. Such peptides can also be made by conventional recombinant techniques using standard procedures described in, for example, Sambrook & Maniatis, Molecular Cloning, A Laboratory Manual. "Recombinant,” as used herein, means that a gene is derived from a recombinant (e.g., microbial or mammalian) expression system that has been genetically modified to contain a polynucleotide encoding a GLP-1 peptide as described herein.
  • a recombinant e.g., microbial or mammalian
  • GLP-1 its agonists, analogs, derivatives, variants, and biologically active fragments, that are peptides may be a naturally purified product, or a product of synthetic chemical procedures, or produced by recombinant techniques from prokaryotic or eukaryotic hosts (for example, by bacteria, yeast, higher plant, insect, or mammalian cells in culture or in vivo). Pepending on the host employed in a recombinant production procedure, the polypeptides of the present invention are generally non-glycosylated, but may be glycosylated.
  • the GLP-1 peptides can be recovered and purified from recombinant cell cultures by methods including, but not limited to, ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography, and lectin chromatography.
  • High- performance liquid chromatography (HPLC) can be employed for final purification steps.
  • compositions of the invention include exendins, which refer to naturally occurring exendin peptides that are found in Gila-monster.
  • exendins include exendin-3 (SEQ IP NO:12), which is present in the salivary secretions of Heloderma harridum, exendin-4 (SEQ IP NO: 14), which is a peptide present in the salivary secretions of Heloderma suspectum (Eng, J., et al, J. Biol. Chem., 265:20259-62, 1990; Eng., J., et al, J. Biol. Chem., 267:7402-05, 1992), and agonists, analogs, derivative, variants of either of them as well as biologically active fragments thereof.
  • Exendin-4 as it occurs in the salivary secretions of the Gila monster, is an amidated peptide.
  • exendin As it occurs in the salivary secretions of the Gila monster, is an amidated peptide.
  • exendin refers to both the amidated form of the peptide and the acid form of the peptide.
  • GLP-1 generally refers to the amidated 7-36 molecule, but it is also intended to include non-amidated molecules.
  • Exendin agonist refers to compounds that mimic any effect of an exendin by binding to the receptor or receptors where a naturally occurring exendin exerts an effect.
  • agonist activity in this context means having a biological activity of an exendin, such as those described herein; but it is understood that the activity of the agonist can be either less potent or more potent than the native exendin.
  • Exendin-4 is a 39-amino acid polypeptide. Certain sequences are compared in Table 1. TABLE 1 a. HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR(NH 2 ) b. HSDGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS(NH 2 ) c. DLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS(NH 2 ) d. HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS(NH 2 ) e. HSDATFTAEYSKLLAKLALQKYLESILGSSTSPRPPSS f. HSDATFTAEYSKLLAKLALQKYLESILGSSTSPRPPS g.
  • HSDAIFTQQYSKLLAKLALQKYLASILGSRTSPPP(NH 2 ) a GLP-1 (7-36) (NH 2 ) [SEQ IP NO: 1].
  • b exendin 3 (NH 2 ) [SEQ IP NO: 12].
  • c exendin 4 (9-39)(NH 2 ) [SEQ IP NO: 13].
  • d exendin 4 (NH 2 ) [SEQ IP NO: 14].
  • e helospectin I [SEQ IP NO: 15].
  • f helospectin II [SEQ IP NO: 16].
  • g helodermin (NH 2 ) [SEQ IP NO: 17].
  • h Q 8 , Q 9 helodermin (NH 2 ) [SEQ IP NO: 18].
  • exendin-4 An insulinotropic action of exendin-4 has also been observed in rodents, improving insulin response to glucose by over 100% in non-fasted Harlan Sprague Pawley (HSP) rats, and by up to ⁇ 10-fold in non-fasted db/db mice. Higher pretreatment plasma glucose concentrations were associated with greater glucose-lowering effects. Thus the observed glucose lowering effect of exendin-4 appears to be glucose-dependent, and minimal if animals are already euglycemic. Pegradation studies with exendin-4 compared to GLP-1 indicate that exendin-4 is relatively resistant to degradation.
  • exendin agonist includes any molecules, whether they be peptides, peptide mimetics, or other chemical compounds, that bind to or activate a receptor or receptors at which exendin exerts an effect, as described above.
  • exendin agonists may include molecules having insulinotropic activity and that may bind a GLP- 1 receptor molecule in in vitro assays and induce second messenger activity on, inter alia, insulin producing ⁇ -cells.
  • exendin The structure activity relationship (SAR) of exendin was investigated for structures that may relate to the activity of exendin, for its stability to metabolism, and for improvement of its physical characteristics, especially as it pertains to peptide stability and to amenability to alternative delivery systems, and various exendin agonist peptide compounds have been invented.
  • Exendm agonists include exendin analogs with agonist activity in which one or more naturally or non-naturally occurring amino acids are added, inserted, eliminated or replaced with another amino acid(s).
  • Preferred exendin analogs are peptide analogs of exendin-4.
  • Exendin analogs include peptides that are encoded by polynucleotides that express biologically active exendin analogs with agonist activity, as defined herein.
  • exendin analogs may be peptides containing one or more amino acid substitutions, extensions, additions or deletions, compared with exendin-4 or exendin-3.
  • the number of substitutions, extension, deletions, or additions is 30 amino acids or less, 25 amino acids or less, 20 amino acids or less, 15 amino acids or less, 10 amino acids or less, 5 amino acids or less or any integer in between these amounts.
  • the substitutions include one or more conservative substitutions.
  • Exendin analogs which include chemically derivatized or altered compounds and peptides having a preferred amino acid homology to SEQ IP NOs:12 and 14 have been previously described and are contemplated to be within the scope of the claimed invention.
  • exendin analogs with agonist activity are described in PCT Application Serial No. PCT US98/00449, filed January 7, 1998, entitled “Use of Exendins and Agonists Thereof for the Reduction of Food Intake," which claims priority to U.S. Provisional Application No. 60/034,905 filed January 7, 1997, both of which are hereby incorporated by reference.
  • Activity as exendin agonists and exendin analogs with agonist activity can be indicated, for example, by activity in the assays incorporated by reference in the referenced applications. Effects of exendins or exendin agonists can be identified, evaluated, or screened for, using the methods described herein, or other art-known or equivalent methods for determining the effects of exendin.
  • Screening assays for potential exendin agonist compounds or candidate exendin agonist compounds may include an in vitro GLP-1 receptor assay/screen described above, an amylin receptor assay/screen using an amylin receptor preparation as described in U.S. Patent No. 5,264,372, issued November 23, 1993, the contents of which are incorporated herein by reference, one or more calcitonin receptor assays/screens using, for example, T47P and MCF7 breast carcinoma cells, which contain calcium receptors coupled to the stimulation of adenyl cyclase activity, and/or a CGRP receptor assay/screen using, for example, SK-N-MC cells.
  • exendin-4 (1-30) [SEQ IP NO:19: His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gin Met Glu Glu Glu Ala Val Arg Leu Phe He Glu Trp Leu Lys Asn Gly Gly]; exendin-4 (1-30) amide [SEQ IP NO:20: His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser
  • compositions including said compounds and salts thereof are also included within the scope of the present invention.
  • Exendin analogs with agonist activity also include those described in U.S. Provisional Application No. 60/065,442, including compounds of the formula (III) [SEQ IP NO:25]: Xaai Xaa 2 Xaa 3 Gly Xaa 5 Xaa 6 Xaa 7 Xaa 8 Xaa 9 aaio Xaa Xaa 12 Xaa 13 Xaa 1 Xaa 15 Xaa ⁇ 6 Xaa 1 Ala Xaa ⁇ 9 Xaa 20 Xaa 2 j Xaa 22 Xaa 23 Xaa 2 Xaa 25 Xaa 6 Xaa 27 Xaa 28 -Z 1 ;
  • Xaai is His, Arg or Tyr
  • Xaa 2 is Ser, Gly, Ala or Thr;
  • Xaa is Asp or Glu
  • Xaa 5 is Ala or Thr
  • Xaa 6 is Ala, Phe, Tyr or naphthylalanine
  • Xaa 7 is Thr or Ser
  • Xaa 8 is Ala, Ser or Thr;
  • Xaa 9 is Asp or Glu
  • Xaaio is Ala, Leu, He, Val, pentylglycine or Met;
  • Xaa ⁇ 2 is Ala or Lys
  • Xaa ⁇ 3 is Ala or Gin
  • Xaa ⁇ is Ala, Leu, He, pentylglycine, Val or Met;
  • Xaa 15 is Ala or Glu
  • Xaai ⁇ is Ala or Glu
  • Xaa 17 is Ala or Glu
  • Xaa ⁇ is Ala or Val
  • Xaa 20 is Ala or Arg
  • Xaa 2 ⁇ is Ala or Leu
  • Xaa 22 is Ala, Phe, Tyr or naphthylalanine
  • Xaa 23 is He, Val, Leu, pentylglycine, tert-butylglycine or Met;
  • Xaa 2 is Ala, Glu or Asp
  • Xaa 25 is Ala, Trp, Phe, Tyr or naphthylalanine
  • Xaa 26 is Ala or Leu
  • Xaa 27 is Ala or Lys
  • Xaa 28 is Ala or Asn
  • N-alkyl groups for N-alkylglycine, N-alkylpentylglycine and N-alkylalanine include lower alkyl groups preferably of 1 to about 6 carbon atoms, more preferably of 1 to 4 carbon atoms.
  • Preferred exendin analogs include those wherein Xaai is His or Tyr. More preferably
  • Preferred compounds are those where Xaa 6 is Phe or naphthylalanine; Xaa 22 is Phe or naphthylalanine and Xaa 23 is He or Val.
  • Xaa 3 ⁇ , Xaa 36 , Xaa 37 and Xaa 38 are independently selected from Pro, homoproline, thioproline and N-alkylalanine.
  • Z x is -NH 2 .
  • Z 2 is -NH 2 .
  • Xaai is His or Tyr, more preferably His;
  • Xaa 2 is Gly;
  • Xaa 6 is Phe or naphthylalanine;
  • Xaan is Leu, pentylglycine or Met;
  • Xaa 22 is Phe or naphthylalanine;
  • Xaa 23 is He or Val;
  • Xaa 3 j, Xaa 36 , Xaa 37 and Xaa 38 are independently selected from Pro, homoproline, thioproline or N-alkylalanine. More preferably Zj is -NH 2 .
  • especially preferred compounds include those of formula (III) wherein: Xaai is His or Arg; Xaa 2 is Gly or Ala; Xaa 3 is Asp or Glu; Xaas is Ala or Thr; Xaa 6 is Ala, Phe or nephthylalaine; Xaa 7 is Thr or Ser; Xaa 8 is Ala, Ser or Thr; Xaa is Asp or Glu; Xaaio is Ala, Leu or pentylglycine; Xaan is Ala or Ser; Xaa J2 is Ala or Lys; Xaaj 3 is Ala or Gin; Xaa ] is Ala, Leu or pentylglycine; Xaais is Ala or Glu; Xaa ⁇ 6 is Ala or Glu; Xaan is Ala or Glu; Xaa ⁇ 9 is Ala or Val; Xaaa
  • Xaa ! is Leu, He, Val or pentylglycine, more preferably Leu or pentylglycine
  • Xaa 2 s is Phe, Tyr or naphthylalanine, more preferably Phe or naphthylalanine.
  • Exendin analogs with agonist activity also include those described in U.S. Provisional Application No. 60/066,029, including compounds of the formula (IV)[SEQ IP NO:26]:
  • Xaai is His, Arg, Tyr, Ala, Norval, Val or Norleu
  • Xaa 2 is Ser, Gly, Ala or Thr
  • Xaa 3 is Ala, Asp or Glu
  • Xaa is Ala, Norval, Val, Norleu or Gly;
  • Xaa 5 is Ala or Thr
  • Xaa 6 is Phe, Tyr or naphthylalanine
  • Xaa 7 is Thr or Ser
  • Xaa 8 is Ala, Ser or Thr;
  • Xaa 9 is Ala, Norval, Val, Norleu, Asp or Glu;
  • Xaaio is Ala, Leu, He, Val, pentylglycine or Met;
  • Xaan is Ala or Ser
  • Xaa ⁇ 2 is Ala or Lys
  • Xaau is Ala, Leu, He, pentylglycine, Val or Met;
  • Xaais is Ala or Glu
  • Xaai ⁇ is Ala or Glu
  • Xaan is Ala or Glu
  • Xaa ⁇ is Ala or Val
  • Xaa 2 o is Ala or Arg
  • Xaa 22 is Phe, Tyr or naphthylalanine
  • Xaa 23 is He, Val, Leu, pentylglycine, tert-butylglycine or Met;
  • Xaa 2 is Ala, Glu or Asp
  • Xaa 25 is Ala, Trp, Phe, Tyr or naphthylalanine
  • Xaa 26 is Ala or Leu
  • Xaa 27 is Ala or Lys
  • Xaa 28 is Ala or Asn
  • Xaa ⁇ , Xaa 36 , Xaa 3 and Xaa 38 are independently Pro,homoproline, 3Hyp, 4Hyp, thioproline, N-alkylglycine, N-alkylpentylglycine or N-alkylalanine; and
  • Z 2 is -OH or -NH 2 ; provided that no more than three of Xaa 3 , Xaa
  • N-alkyl groups for N-alkylglycine, N-alkylpentylglycine and N-alkylalanine include lower alkyl groups preferably of 1 to about 6 carbon atoms, more preferably of 1 to 4 carbon atoms.
  • Suitable compounds of formula (II) include those described in application Serial No. PCT/US98/24273, filed November 13, 1998, entitled “Novel Exendin Agonist Compounds", identified therein in Examples 1-89 ("Compounds 1-89,” respectively), as well as those corresponding compounds identified therein in Examples 104 and 105.
  • Preferred such exendin analogs include those wherein Xaai is His, Ala or Norval. More preferably Xaai is His or Ala. Most preferably Xaai is His.
  • Preferred compounds of formula (IN) are those where Xaa 6 is Ala, Phe or naphthylalanine; Xaa 2 is Phe or naphthylalanine; and Xaa 23 is He or Val.
  • Xaa 3 ⁇ , Xaa 36 , Xaa 7 and Xaa 38 are independently selected from Pro, homoproline, thioproline and ⁇ -alkylalanine.
  • Zi is - ⁇ H 2 .
  • Z 2 is -NH 2 .
  • Xaai is Ala, His or Tyr, more preferably Ala or His
  • Xaa 2 is Ala or Gly
  • Xaa 6 is Phe or naphthylalanine
  • Xaaj 4 is Ala, Leu, pentylglycine or Met
  • Xaa 22 is Phe or naphthylalanine
  • Xaa 23 is He or Val
  • Xaa 3 ⁇ , Xaa 36 , Xaa 37 and Xaa 38 are independently selected from Pro, homoproline, thioproline or N-alkylalanine
  • Xaa 39 is Ser or Tyr, more preferably Ser. More preferably Z-* is -NH 2 .
  • especially preferred compounds include those of formula (IN) wherein: Xaai is His or Ala; Xaa 2 is Gly or Ala; Xaa 3 is Ala, Asp or Glu; Xaa is Ala or Gly; Xaa 5 is Ala or Thr; Xaa 6 is Phe or naphthylalanine; Xaa 7 is Thr or Ser; Xaa 8 is Ala, Ser or Thr; Xaa 9 is Ala, Asp or Glu; Xaaio is Ala, Leu or pentylglycine; Xaan is Ala or Ser; Xaaj 2 is Ala or Lys; Xaa ⁇ 3 is Ala or Gin; Xaa ⁇ 4 is Ala, Leu, Met or pentylglycine; Xaais is Ala or Glu; Xaa ⁇ 6 is Ala or Glu; Xaan is Ala or Glu; Xaan is Ala or
  • Xaa ⁇ is Ala, Leu, He, Val or pentylglycine, more preferably Leu or pentylglycine
  • Xaa 25 is Ala, Phe, Tyr or naphthylalanine, more preferably Phe or naphthylalanine.
  • narrower genera of compounds having peptides of various lengths for example genera of compounds which do not include peptides having a length of 28, 29 or 30 amino acid residues, respectively.
  • present invention includes narrower genera of compounds described in PCT application Serial No. PCT/US98/24210, filed November 13, 1998, entitled “Novel Exendin Agonist Compounds" and having particular amino acid sequences, for example, compounds of the formula (V) [SEQ. IP. NO:27]:
  • Xaai Xaa 2 Xaa 3 Gly Xaa 5 Xaa 6 Xaa 7 Xaa 8 Xaa 9 Xaaio Xaan Xaa ⁇ 2 Xaa ⁇ 3 Xaaj Xaa ⁇ Xaa ⁇ 6 Xaan Ala Xaa ⁇ 9 Xaa 20 Xaa 2 ⁇ Xaa 22 Xaa 23 Xaa 2 Xaa 2 s Xaa 26 Xaa 2 Xaa 28 -Z ⁇ ;
  • Xaai is His or Arg
  • Xaa 2 is Gly or Ala
  • Xaa 3 is Asp or Glu
  • Xaa 5 is Ala or Thr
  • Xaa 6 is Ala, Phe or naphthylalanine
  • Xaa 7 is Thr or Ser
  • Xaa 8 is Ala, Ser or Thr
  • Xaa 9 is Asp or Glu
  • Xaaio is Ala, Leu or pentylglycine
  • Xaan is Ala or Ser
  • Xaa ⁇ 2 is Ala or Lys; Xaa ] is Ala or Gin;
  • Xaa ⁇ is Ala, Leu or pentylglycine
  • Xaai 5 is Ala or Glu
  • Xaan is Ala or Glu; Xaa ⁇ 9 is Ala or Val;
  • Xaa 2 o is Ala or Arg
  • Xaa 2 ⁇ is Ala r Leu
  • Xaa 22 is Phe or naphthylalanine
  • Xaa 23 is He, Val or tert-butylglycine;
  • Xaa 2 is Ala, Glu or Asp;
  • Xaa 25 is Ala, Trp, or Phe;
  • Xaa 26 is Ala or Leu
  • Xaa 27 is Ala or Lys
  • Xaa 28 is Ala or Asn; Zi is -OH,
  • Xaa ⁇ , Xaa 6 , Xaa 3 and Xaa 38 are independently selected from the group consisting of Pro, homoproline, thioproline and N-methylylalanine; and Z 2 is -OH or -NH 2 ; provided that no more than three of Xaa 3 , Xaa 5 , Xaag, Xaa 8 , Xaaio, Xaan, Xaa ⁇ 2 , Xaaj 3 , Xaa ] , Xaais, Xaa ⁇ 6 , Xaan, X a ⁇ , Xaa 2 o, Xaa 2 ⁇ , Xaa 2 , Xaa 2 s, Xaa 26 , Xaa 27 and Xaa 28 are Ala; and pharmaceutically acceptable salts thereof.
  • the present invention includes narrower genera of peptide compounds described in PCT Application Serial No. PCT/US98/24273, filed November 13, 1998, entitled "Novel Exendin Agonist Compounds" as having particular amino acid sequences, for example, compounds of the formula [VT] [SEQ. IP. NO:28]:
  • Xaan Xaa ⁇ 2 Xaa ⁇ 3 Xaan Xaa ⁇ 5 Xaa ⁇ 6
  • Xaa 7 is Thr or Ser
  • Xaa 8 is Ala, Ser or Thr;
  • Xaa 9 is Ala, Asp or Glu
  • Xaaio is Ala, Leu or pentylglycine
  • Xaan is Ala or Ser
  • Xaa ⁇ 2 is Ala or Lys
  • Xaa ⁇ 3 is Ala or Gin
  • Xaa ⁇ 4 is Ala, Leu, Met or pentylglycine
  • Xaai 6 is Ala or Glu
  • Xaan is Ala or Glu
  • Xaa ⁇ 9 is Ala or Val
  • Xaa 20 is Ala or Arg
  • Xaa 2 ⁇ is Ala or Leu
  • Xaa 22 is Phe or naphthylalanine
  • Xaa 23 is He, Val or tert-butylglycine
  • Xaa 24 is Ala, Glu or Asp
  • Xaa 25 is Ala, Trp or Phe;
  • Xaa 26 is Ala or Leu
  • Xaa 27 is Ala or Lys
  • Xaa 28 is Ala or Asn
  • Xaa 3 ⁇ , Xaa 36 , Xaa 37 and Xaa 38 are independently Pro,homoproline, thioproline, or N-methylylalanine;
  • Z 2 is -OH or -NH 2 ; provided that no more than three of Xaa 3 , Xaa 5 , Xaa 6 , Xaa 8 , Xaaio, Xaan, aa ⁇ 2 , Xaa ⁇ 3 ,
  • Xaa ⁇ 4 , Xaais, Xaa 16 , Xaan, Xaa ⁇ , Xaa 2 o, Xaa 2 ⁇ , Xaa 2 , Xaa 2 s, Xaa 26 , Xaa 27 , and Xaa 28 are Ala; and provided that, if Xaai is His, Arg or Tyr, then at least one of Xaa 3 , Xaa and Xaa 9 is Ala; and pharmaceutically acceptable salts thereof.
  • Preferred compounds of formula (VI) include those wherein Xaai is His, Ala, Norval or 4-imidazopropionyl.
  • Xaai is His, or 4-imidazopropionyl or Ala, more preferably His or 4-imidazopropionyl.
  • Preferred compounds of formula (VI) include those wherein Xaa2 is Gly.
  • Preferred compounds of formula (VT) include those wherein Xaa4 is Ala.
  • Preferred compounds of formula (VT) include those wherein Xaa9 is Ala.
  • Preferred compounds of formula (VI) include those wherein Xaal4 is Leu, pentylglycine or Met.
  • Preferred compounds of formula (VT) include those wherein Xaa25 is Trp or Phe.
  • Preferred compounds of formula (VI) include those wherein Xaa6 is Ala, Phe or naphthylalanine; Xaa22 is Phe or naphthylalanine; and Xaa23 is He or Val.
  • Preferred compounds of formula (VI) include those wherein Zl is -NH2.
  • Preferred compounds of formula (VI) include those wherein Xaa31, Xaa36, Xaa37 and Xaa38 are independently selected from the group consisting of Pro, homoproline, thioproline and N-alkylalanine.
  • Preferred compounds of formula (VI) include those wherein Xaa39 is Ser or Tyr, preferably Ser.
  • Preferred compounds of formula (VI) include those wherein Z2 is -NH2.
  • Preferred compounds of formula (VI) include those 42 wherein Zl is -NH2.
  • Preferred compounds of formula (VI) include those wherein Xaa21 is Lys-NH2-R where R is Lys, Arg, C1-C10 straight chain or branched alkanoyl.
  • Preferred compounds of formula (VI) include those wherein XI is Lys Asn, Lys-NH ⁇ -R
  • Xaai is His, Arg or Tyr or 4-imidazopropionyl
  • Xaa 2 is Ser, Gly, Ala or Thr;
  • Xaa 3 is Asp or Glu
  • Xaa 5 is Ala or Thr
  • Xaa 6 is Ala, Phe, Tyr or naphthylalanine
  • Xaa 7 is Thr or Ser
  • Xaa 8 is Ala, Ser or Thr;
  • Xaa 9 is Asp or Glu
  • Xaaio is Ala, Leu, He, Val, pentylglycine or Met; Xaai i is Ala or Ser;
  • Xaa ⁇ 2 is Ala or Lys
  • Xaa ⁇ 3 is Ala or Gin
  • Xaa ⁇ 4 is Ala, Leu, He, pentylglycine, Val or Met;
  • Xaan is Ala or Glu
  • Xaa 2 o is Ala or Arg
  • Xaa 2 is Ala, Leu or Lys-NH ⁇ -R where R is Lys, Arg, Ci-Cio straight chain or branched alkanoyl or cycloalkylalkanoyl;
  • Xaa 22 is Phe, Tyr or naphthylalanine
  • Xaa 23 is He, Val, Leu, pentylglycine, tert-butylglycine or Met;
  • Xaa 24 is Ala, Glu or Asp
  • Xaa 25 is Ala, Trp, Phe, Tyr or naphthylalanine;
  • Xaa 26 is Ala or Leu;
  • Xi is Lys Asn, Asn Lys, Lys-NH ⁇ -R Asn, Asn Lys-NH ⁇ -R, Lys-NH ⁇ -R Ala, Ala Lys- NH ⁇ -R where R is Lys, Arg, Ci-Cio straight chain or branched alkanoyl or cycloalkylalkanoyl Zi is -OH, -NH 2 ,
  • Gly-Z 2 Gly Gly-Z 2 , Gly Gly-Z 2 , Gly Gly Xaa 3 ⁇ -Z 2 , Gly Gly Xaa 3 ⁇ Ser-Z 2 , Gly Gly Xaa 3 ⁇ Ser Ser-Z 2 ,
  • Xaa 3 ⁇ , Xaa 36 , Xaa 37 and Xaa 8 are independently selected from the group consisting of Pro, homoproline, 3Hyp, 4Hyp, thioproline, N-alkylglycine, N-alkylpentylglycine and N-alkylalanine; and
  • Z 2 is -OH or -NH 2 ; provided that no more than three of Xaa 3 , Xaa 5 , Xaa 6 , Xaa 8 , Xaaio, Xaan, Xaa ⁇ 2 , Xaa ⁇ 3 ,
  • Xaa ⁇ , Xaais, Xaa ⁇ 6 , Xaan, Xaa ⁇ 9 , Xaa 2 o, Xaa 2 ⁇ , Xaa 24 , Xaa 25 , and Xaa 26 are Ala.
  • pharmaceutically acceptable salts of the compound of formula (VII) and pharmaceutical compositions including said compounds and salts thereof are also within the scope of the present invention.
  • Preferred exendin analogs of formula (VII) include those wherein Xaai is His, Tyr or 4- imidazopropionyl. More preferably Xaai is His.
  • Xaa 6 is Phe or naphthylalanine
  • Xaa 22 is Phe or naphthylalanine
  • Xaa 23 is He or Val.
  • Zi is -NH 2 .
  • especially preferred are such compounds of formula (VII) wherein Xaa 3 ⁇ , Xaa 36 , Xaa 37 and Xaa 38 are independently selected from the group consisting of Pro, homoproline, thioproline and N-alkylalanine. More preferds, Z 2 is -NH 2 .
  • Preferred compounds of formula (VII) include those wherein Xi is Lys Asn, Lys-NH ⁇ -R Asn, or Lys-NH ⁇ -R Ala where R is Lys, Arg, Ci-Cio straight chain or branched alkanoyl.
  • Preferred compounds of formula (VII) include compounds described in PCT application Serial No. PCT/US98/24210, filed November 13, 1998, entitled “Novel Exendin Agonist Compounds" and identified therein as Compound Nos. 62-69.
  • exendin analogs include those wherein Xaai is His, Ala or Norval. More preferably Xaai is His or Ala. Most preferably Xaai is His. Preferred are those compounds of formula (VII) wherein Xaa 2 is Gly.
  • Preferred compounds of formula (VII) are those wherein Xaa 2 s is Trp or Phe.
  • Preferred compounds of formula (VII) are those where Xaa 6 is Ala, Phe or naphthylalanine; Xaa 22 is Phe or naphthylalanine; and Xaa 23 is He or Val.
  • Zi is -NH 2 .
  • Z 2 is -NH 2 .
  • Xaai is Ala, His or Tyr, more preferably Ala or His
  • Xaa 2 is Ala or Gly
  • Xaa 6 is Phe or naphthylalanine
  • Xaa J is Ala, Leu, pentylglycine or Met
  • Xaa 22 is Phe or naphthylalanine
  • Xaa 23 is He or Val
  • Xaa 3 ⁇ , Xaa 36 , Xaa 7 and Xaa 38 are independently selected from Pro, homoproline, thioproline or N-alkylalanine
  • Xaa 3 is Ser or Tyr, more preferably Ser. More preferably Zi. is -NH 2 .
  • especially preferred compounds include those of formula (VII) wherein: Xaai is His or Ala; Xaa 2 is Gly or Ala; Xaa 3 is Ala, Asp or Glu; Xaa 4 is Ala or Gly; Xaa 5 is Ala or Thr; Xaa 6 is Phe or naphthylalanine; Xaa 7 is Thr or Ser; Xaa 8 is Ala, Ser or Thr; Xaa 9 is Ala, Asp or Glu; Xaaio i Ala, Leu or pentylglycine; Xaan is Ala or Ser; Xaa ⁇ 2 is Ala or Lys; Xaa ⁇ 3 is Ala or Gin; Xaa ⁇ is Ala, Leu, Met or pentylglycine; Xaa ⁇ is Ala or Glu; Xaa i6 is Ala or Glu; Xaan is Ala or Glu
  • Especially preferred compounds of formula (VII) include those described in PCT application Serial No. PCT/US98/24210, filed November 13, 1998, entitled “Novel Exendm Agonist Compounds” and having the amino acid sequences identified therein as SEQ. IP. NOS. 5-93.
  • Xaai 4 is Ala, Leu, He, Val or pentylglycine, more preferably Leu or pentylglycine
  • Xaa25 is Ala, Phe, Tyr or naphthylalanine, more preferably Phe or naphthylalanine.
  • Xaai is His, Arg, Tyr, Ala, Norval, Val, Norleu or 4-imidazopropionyl
  • Xaa 2 is Ser, Gly, Ala or Thr;
  • Xaa 8 is Ala, Ser or Thr;
  • Xaa 9 is Ala, Norval, Val, Norleu, Asp or Glu;
  • Xaaio is Ala, Leu, He, Val, pentylglycine or Met;
  • Xaan is Ala or Ser
  • Xaa ⁇ 2 is Ala or Lys
  • Xaa ⁇ 3 is Ala or Gin
  • Xaa ⁇ 4 is Ala, Leu, He, pentylglycine, Val or Met;
  • Xaai 6 is Ala or Glu:
  • Xaan is Ala or Glu
  • Xaa ⁇ is Ala or Val
  • Xaa 2 o is Ala or Arg
  • Xaa 2 ⁇ is Ala, Leu or Lys-NH -R where R is Lys, Arg, C ⁇ ,1- " 10 straight chain or branched alkanoyl or cycloalleyl-alkanoyl;
  • Xaa 22 is Phe, Tyr or naphthylalanine
  • Xaa 23 is He, Val, Leu, pentylglycine, tert-butylglycine or Met;
  • Xaa 24 is Ala, Glu or Asp;
  • Xaa 25 is Ala, Trp, Phe, Tyr or naphthylalanine;
  • Xaa 26 is Ala or Leu;
  • Xi is Lys Asn, Asn Lys, Lys-NH ⁇ -R Asn, Asn Lys-NH ⁇ -R, Lys-NH ⁇ -R Ala, Ala Lys- NH ⁇ -R where R is Lys, Arg, Ci-Cio straight chain or branched alkanoyl or cycloalkylalkanoyl
  • Xaa 31 , Xaa 36 , Xaa 37 and Xaa 8 are independently selected from the group consisting of Pro, homoproline, 3Hyp, 4Hyp, thioproline, N-alkylglycine, N-alkylpentylglycine and N-alkylalanine; and
  • Z 2 is -OH or-NH 2 ; provided that no more than three of Xaa 3 , Xaa , Xaas, Xaa 6 , Xaa 8 , Xaa 9 , Xaaio, X an, Xaa ⁇ 2 , Xaa ⁇ 3
  • Xaai is His, or 4-imidazo ⁇ ropionyl or Ala, more preferably His or 4-imidazopropionyl.
  • Preferred compounds of formula (VIII) include those wherein Xaa 2 is Gly.
  • Preferred compounds of formula (NIII) include those wherein Xaa 4 is Ala.
  • Preferred compounds of formula (VIII) include those wherein Xaa 9 is Ala.
  • Preferred compounds of formula (VIII) include those wherein Xaa ⁇ is Leu, pentylglycine or Met.
  • Preferred compounds of formula (VIII) include those wherein Xaa 25 is Trp or Phe.
  • Preferred compounds of formula (VTJI) include those wherein Xaa 6 is Ala, Phe or naphthylalanine; Xaa 22 is Phe or naphthylalanine; and Xaa 23 is He or Val.
  • Preferred compounds of formula (VIII) include those wherein Zi is - ⁇ H 2 .
  • Preferred compounds of formula (VIII) include those wherein Xaa 3 ⁇ , Xaa 6 , Xaa 3 and Xaa 38 are independently selected from the group consisting of Pro, homoproline, thioproline and N-alkylalanine.
  • Preferred compounds of formula (VIII) include those wherein Xaa 39 is Ser or Tyr, preferably Ser.
  • Preferred compounds of formula (VIII) include those wherein Z 2 is -NH 2 .
  • Preferred compounds of formula (VIII) include those 42 wherein Zj is -NH 2 .
  • Preferred compounds of formula (VIII) include those wherein Xaa 2 ⁇ is Lys-NH ⁇ -R where R is Lys, Arg, -Cio straight chain or branched alkanoyl.
  • Preferred compounds of formula (VIII) include those wherein Xi is Lys Asn, Lys-NH ⁇ -R Asn, or Lys-NH ⁇ -R Ala where R is Lys, Arg, Ci-Cio straight chain or branched alkanoyl.
  • Preferred compounds of formula (VTII) include those described in PCT Application Serial No. PCT/US98/24273, filed November 13, 1998, entitled “Novel Exendin Agonist Compounds" as having an amino acid sequence selected from those identified therein as SEQ. IP. NOS. 95-110.
  • Xaa 2 is Ser, Gly, Ala or Thr;
  • Xaa 3 is Asp or Glu
  • Xaa is Phe, Tyr or naphthalanine
  • Xaa 5 is Thr or Ser
  • Xaa 6 is Ser or Thr
  • Xaa 7 is Asp or Glu
  • Xaa 8 is Leu, He, Val, pentylglycine or Met;
  • Xaa 9 is Leu, He, pentylglycine, Val or Met;
  • Xaaio is Phe, Tyr or naphthalanine
  • Xaai l is He, Val, Leu, pentylglycine, tert-butylglycine or Met;
  • Xaa ⁇ 2 is Glu or Asp
  • Xaa ⁇ 3 is Trp, Phe, Tyr, or naphthylalanine
  • Xaa 1 , Xaa ⁇ , Xaa ⁇ 6 and Xaan are independently Pro, homoproline, 3Hyp, 4Hyp, thioproline, N-alkylglycine, N-alkylpentylglycine or N-alkylalanine;
  • Xaais is Ser, Thr or Tyr; and Z is -OH or -NH 2 ; with the proviso that the compound does not have the formula of either SEQ. IP.
  • N-alkyl groups for N-alkylglycine, N-alkylpentylglycine and N- alkylalanine include lower alkyl groups preferably of 1 to about 6 carbon atoms, more preferably of 1 to 4 carbon atoms.
  • pharmaceutically acceptable salts of the compounds of formula (IX) include those wherein Xaai is His or Tyr. More preferably
  • Xaa is Phe or naphthalanine
  • Xaan is He or Val
  • Xaa ⁇ , Xaais, Xaa ⁇ 6 and Xaan are independently selected from Pro, homoproline, thioproline or N-alkylalanine.
  • N-alkylalanine has a N-alkyl group of 1 to about 6 carbon atoms.
  • Xaais, Xaa ⁇ 6 and Xaan are the same amino acid reside.
  • Z is -NH 2 .
  • Xaai is His or Tyr, more preferably His
  • Xaa 2 is Gly
  • Xaa 4 is Phe or naphthalanine
  • Xaa 9 is Leu, pentylglycine or Met
  • Xaaio is Phe or naphthalanine
  • Xaan is He or Val
  • Xaaj , Xaais, Xaa ⁇ and Xaan are independently selected from Pro, homoproline, thioproline or N-alkylalanine
  • Xaais is Ser or Tyr, more preferably Ser. More preferably Z is -NH 2 .
  • especially preferred compounds include those of formula (LX) wherein: Xaai is His or Arg; Xaa 2 is Gly; Xaa 3 is Asp or Glu; Xaa 4 is Phe or napthylalanine; Xaa 5 is Thr or Ser; Xaa 6 is Ser or Thr; Xaa 7 is Asp or Glu; Xaa 8 is Leu or pentylglycine; Xaa 9 is Leu or pentylglycine; Xaaio is Phe or naphthylalanine; Xaan is He, Val or t-butyltylglycine; Xaa ⁇ 2 is Glu or Asp; Xaa i3 is Trp or Phe; Xaa !
  • Xaais, Xaa J6 , and Xaan are independently Pro, homoproline, thioproline, or N-methylalanine;
  • Xaais is Ser or Tyr: and
  • Z is - OH or -NH 2 ; with the proviso that the compound does not have the formula of either SEQ. IP. NOS. 7 or 9. More preferably Z is -NH 2 ..
  • Xaa 9 is Leu, He, Val or pentylglycine, more preferably Leu or pentylglycine
  • Xaa ⁇ 3 is Phe, Tyr or naphthylalanine, more preferably Phe or naphthylalanine.
  • Xaai is His, Arg, Tyr or 4-imidazopropionyl
  • Xaa 2 is Ser, Gly, Ala or Thr;
  • Xaa 3 is Asp or Glu
  • Xaa 4 is Phe, Tyr or naphthylalanine
  • Xaa 5 is Thr or Ser
  • Xaa 6 is Ser or Thr
  • Xaa is Asp or Glu
  • Xaa 8 is Leu, He, Val, pentylglycine or Met
  • Xaa is Leu, He, pentylglycine, Val or Met
  • Xaaio is Phe, Tyr or naphthylalanine
  • Xaan is He, Val, Leu, pentylglycine, tert-butylglycine or Met;
  • Xaai2 is Glu or Asp
  • Xaa ⁇ 3 is Trp, Phe, Tyr, or naphthylalanine;
  • Xj . is Lys Asn, Asn Lys, Lys-NH ⁇ -R Asn, Asn Lys-NH ⁇ -R where R is Lys, Arg, -Cio straight chain or branched alkanoyl or cycloalkylalkanoyl;
  • Xaa ⁇ 4 , Xaais, Xaa ⁇ 6 and Xaan are independently Pro, homoproline, 3Hyp, 4Hyp, thioproline, N-alkylglycine, N-alkylpentylglycine or N-alkylalanine;
  • Xaais is Ser, Thr or Tyr; and Z is -OH or -NH 2 ; with the proviso that the compound does not have the formula of either SEQ. IP. NOS. 7 or 9.
  • Suitable compounds of formula (X) include compounds described in PCT Application Serial No. PCT/US98/16387, filed August 6, 1998, entitled "Novel Exendin Agonist Compounds" having the amino acid sequences of SEQ. IP. NOS. 37-40 therein.
  • Preferred exendin analogs of formula (X) include those wherein Xaai is His, Tyr or 4- imidazopropionyl. More preferably, Xaai is His or 4-imidazopropionyl.
  • Xi is Lys Asn, or Lys-NH ⁇ -R Asn, where R is Lys, Arg, Ci-Cio straight chain or branched alkanoyl.
  • Xaa 4 is Phe or naphthylalanine
  • Xaaio is Phe or naphthylalanine; Xaan is He or Val and Xaa ⁇ , Xaa ⁇ , Xaa ⁇ 6 and Xaan are independently selected from Pro, homoproline, thioproline or N-alkylalanine.
  • Xaa ⁇ 8 is Ser or Tyr. Preferred are those such compounds wherein Xaais is Ser.
  • Z is -NH 2 .
  • Exendins and exendin agonists that are peptides, such as exendm analogs, described herein may be prepared through peptide purification as described in, for example, Eng, et al, J. Biol. Chem. 265:20259-62, 1990; and Eng, et al, J. Biol. Chem. 267:7402-05, 1992, hereby incorporated by reference herein.
  • exendins and exendin agonists that are peptides may be prepared by methods known to those skilled in the art, for example, as described in Raufman, et al, J. Biol. Chem. 267:21432-37, 1992), hereby incorporated by reference herein, using standard solid-phase peptide synthesis techniques and preferably an automated or semiautomated peptide synthesizer as previously described and is well known in the art.
  • Exendins and exendm agonists that are peptides may also be prepared using recombinant PNA techniques, using methods now known in the art. See, e.g., Sambrook et al, Molecular Cloning: A Laboratory Manual, 2d Ed., Cold Spring Harbor (1989). Alternatively, such compounds may be prepared by homogeneous phase peptide synthesis methods. Non-peptide compounds useful in the present invention may be prepared by art-known methods. For example, phosphate-containing amino acids and peptides containing such amino acids, may be prepared using methods known in the art. See, e.g., Bartlett and Landen, Biorg. Chem. 14:356- 377 (1986).
  • Methods for making and/or purifying GLP-1 and its agonists, analogs, derivatives, variants, and fragments can also be utilized to make and/or purify exendins, their agonists, analogs, derivatives, variants, and fragments thereof.
  • compositions of the present invention may be used in combination with a suitable pharmaceutical carrier.
  • Such compositions comprise a therapeutically effective amount of the polypeptide, and a pharmaceutically acceptable carrier or excipient.
  • the compositions of this invention can be administered in any effective, pharmaceutically acceptable form for warm blooded animals, including human and other animal subjects, e.g., in topical, lavage, oral, suppository, parenteral, or infusible dosage forms, as a topical, buccal, sublmgual, pulmonary, or nasal spray or in any other manner effective to deliver the agents.
  • the route of administration will preferably be designed to optimize delivery and/or localization of the agents.
  • the pharmaceutical composition may contain suitable excipients and auxiliaries that facilitate processing of the active compounds into preparations which can be used pharmaceutically.
  • Oral dosage forms encompass tablets, capsules, granules, solutions, and suspensions. Preparations that can be administered rectally include suppositories. Oilier dosage forms include suitable solutions for administration parenterally or orally, and compositions which can be administered buccally or sublmgually.
  • the pharmaceutical preparations of the present invention are manufactured in a manner which is itself well known in the art.
  • the pharmaceutical preparations may be made by means of conventional mixing, granulating, dissolving, and lyophilizing processes.
  • the processes to be used will depend ultimately on the physical properties of the active ingredient used.
  • Suitable formulations for parenteral administration include aqueous solutions of active compounds in water-soluble or water-dispersible form.
  • suspensions of the active compounds as appropriate oily injection suspensions may be administered.
  • Suitable lipophilic solvents or vehicles include fatty oils for example, sesame oil, or synthetic fatty acid esters, for example, ethyl oleate or triglycerides.
  • Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, including for example, sodium carboxymethyl cellulose, sorbitol and/or dextran.
  • Such compositions may also comprise adjuvants such as preserving, wetting, emulsifying, and dispensing agents.
  • compositions may also be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating sterilizing agents into the compositions. They can also be manufactured in the form of sterile solid compositions that can be dissolved or suspended in sterile water, saline, or other injectable medium prior to administration.
  • active ingredients may be administered by a variety of specialized delivery drug techniques that are known to those of skill in the art, such as portable infusion pumps.
  • compositions of the present invention can be administered along with a pharmaceutically acceptable carrier in an amount sufficient to prevent arrhythmias and/or treat an active arrhythmia.
  • a pharmaceutically acceptable carrier in an amount sufficient to prevent arrhythmias and/or treat an active arrhythmia.
  • the compounds of this invention have extremely low toxicity and a low degree of side effects even at high doses.
  • the dosing range of the compounds of this invention will vary depending on a number of factors, such as whether it is used for prophylaxis or treatment of arrhythmia, route of administration, desired dosing schedule, the physical health of the patient, etc.
  • exemplary dose ranges for use in the invention can include 0.001 pmol/kg to 500 nmol/kg per day depending on the composition selected.
  • a lower limit of a dosage range can be about 0.001 pmol/kg, 0.01 pmol/kg, 0.1 pmol/kg, 1 pmol/kg, 10 pmol/kg, or 100 pmol/kg.
  • An upper dosage range can be about 10 pmol/kg, 100 pmol/kg, 1 nmol/kg, 10 nmol/kg, 100 nmol/kg, 250 nmol/kg or 500 nmol/kg.
  • the desired dose will vary depending on the selected active composition.
  • the desired dose will also depend upon other factors including the route of administration and the formulation. For example, continuous infusion as well as bolus doses and sustained release formulations are contemplated. Routes of administration include intramuscular, intravenous, subcutaneous, intradermal, transdermal, intraarticular, intrathecal and the like. Mucosal delivery is also contemplated. These routes include, but are not limited to, oral, nasal, sublingual, rectal, pulmonary and buccal routes, which may include administration of the peptide in liquid, semi- solid or solid form.
  • Exemplary doses for continuous infusion by intravenous (IN.) can be about 0.1 pmol/kg/min to 10 pmol/kg/min and by subcutaneous (s.c.) about 0.1 pmol/kg/min to 75 pmol/kg/min., and for single injection (bolus) by IN. about 0.1 nmol/kg to 2.0 nmol/kg and s.c. about 0.1 nmol/kg to 100 nmol/kg.
  • the foregoing doses may be administered as a single dose or may be divided into multiple doses for administration.
  • the peptides of this invention may be administered once to several times daily.
  • an exemplary dosing rate can be within a range of from about 1 to about 10 pmol/kg per minute of GLP-1 delivered by sustained release subcutaneous, intramuscular, interperitoneal, injected depot with sustained release, deep lung insufflation, as well as by intravenous, buccal, patch or other sustained release delivery methods.
  • Pegradation-resistant GLP-1 analogs, derivatives or variants, exendins, analogs, derivatives or variants, and other molecules of the invention need not be delivered continuously, but are suitable for bolus or sustained release dosing and may be at doses much lower than those described.
  • compositions of the invention which are compatible with the carrier ingredients may also be incorporated into the pharmaceutical formulations.
  • Such drugs may be readily ascertained by those of ordinary skill in the art and may include, for instance, anti- inflammatory agents, diuretics, vasodilators, etc.
  • the present invention contemplates the use of not only the above- stated active forms of the compositions of the invention, but also includes the prodrugs (preforms) which metabolize to the compound and biologically active salt forms thereof, as well as optical isomers which provide the same pharmaceutical results.
  • compositions of the invention may also be used in combination with agents known in the art that enhance the half-life in vivo of peptide in order to enhance or prolong the biological activity of the peptide.
  • agents known in the art that enhance the half-life in vivo of peptide in order to enhance or prolong the biological activity of the peptide.
  • a molecule or chemical moiety may be covalently linked to the composition of the present invention before administration thereof.
  • the enhancing agent may be administered concurrently with the composition.
  • the agent may comprise a molecule that is known to inhibit the enzymatic degradation of the compositions of the invention that may be administered concurrently with or after administration of the composition.
  • Such a molecule may be administered, for example, orally, by injection, or any other means known in the art.
  • compositions of the invention in combination with a pharmaceutically acceptable carrier are preferably administered within the first four hours following an ischemic event in order to prevent the occurrence of cardiac arrhythmia.
  • Compositions of the invention can be co-administered with glucose (5%) if required to maintain blood glucose levels > 5 mM (to maintain efficient insulin secretion).
  • co- administration of potassium (K + ) may be considered, depending on the extent to which activation of the membrane Na + /K + ATPase leads to a shift of K into the intracellular space.
  • compositions of the invention should be commenced concurrently or as soon as possible following therapies that reestablish flow in an artery that was obstructed by a blood clot (e.g., thromolytic therapy) or other obstructive materials, or following an intervention, such as angioplasty, coronary bypass grafting, or placement of an intracoronary stent. Therapy should continue thereafter.
  • the treatment should preferably commence 12-24 hours prior to surgery, during surgery from the onset of anesthesia until aortic crossclamping, and immediately after unclamping for a period of at least 72 hours postoperatively.
  • co-administration of a free radical scavenger or antioxidants will further aid reperfusion recovery.

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Abstract

L'invention concerne des compositions renfermant des composés qui se lient à une incrétine, à un peptide-1 de type glucagone (GLP-1), à une exendine, ou bien à une variante de l'un quelconque des composés susmentionnés. Ces compositions s'utilisent pour la prévention et le traitement d'arythmies associées à une ischémie cardiaque, à une perfusion ischémique cardiaque ou à une insuffisance cardiaque congestive. L'invention porte à la fois sur la méthode et sur les compositions de traitement.
EP03813770A 2002-12-17 2003-12-17 Prevention et traitement d'arythmies cardiaques Withdrawn EP1610811A4 (fr)

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TW201609795A (zh) 2013-12-13 2016-03-16 賽諾菲公司 作為雙重ｇｌｐ－１／ｇｉｐ受體促效劑的艾塞那肽－４（ｅｘｅｎｄｉｎ－４）胜肽類似物
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CN108939052B (zh) * 2018-09-04 2021-03-16 江苏省中医药研究院 艾塞那肽在制备预防或治疗房颤的药物中的用途
CN110551203B (zh) * 2019-09-25 2023-02-10 成都奥达生物科技有限公司 一种艾塞那肽类似物
IL294521A (en) 2020-02-18 2022-09-01 Novo Nordisk As glp-1 compounds and their uses

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EP1610811A4 (fr) 2008-03-26
AU2003297356A1 (en) 2004-07-14
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AU2003297356A8 (en) 2004-07-14
WO2004056313A2 (fr) 2004-07-08

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