JP2006520747A - Methods and compositions for treating polycystic ovary syndrome - Google Patents

Abstract

The present invention relates to a method for treating polycystic ovary syndrome (PCOS), characterized by administering exendin or an analog or agonist thereof to a subject suffering from polycystic ovary syndrome.

Description

Detailed Description of the Invention

This application is a continuation-in-part of U.S. Application No. 10 / 317,126, filed on December 11, 2002, and U.S. Application No. 10 / 317,126, filed on Dec. 11, 2002. Is a continuation-in-part of International Application No. PCT / US03 / 01109 and claims priority to US Provisional Application No. 60 / 350,395 filed on January 22, 2002 .

The present invention relates to endocrinology and pharmacology. In particular, it relates to methods and compositions for treating subjects suffering from polycystic ovary syndrome (PCOS).

BACKGROUND OF THE INVENTION Polycystic ovary syndrome (PCOS), also known as polycystic ovary or Stein-Lebental syndrome, affects an estimated 6-10% of women in the United States. PCOS is characterized by anovulation (irregular or menstrual missing) and hyperandrogen (high serum testosterone and androstenedione). Further etiological and clinical symptoms of this disease are abnormal uterine bleeding, enlarged polyfollicular ovary, infertility, obesity, insulin resistance, hyperinsulinemia, hypertension, hyperlipidemia, type 2 diabetes, excessive May include facial hair growth, hair loss and acne.

  Insulin resistance and hyperinsulinemia have a high prevalence in patients with PCOS and may clearly indicate the pathophysiology of this disease (Udoff, L., et al., Curr. Opin. Obstret. Gynecol 7: 340-343 (1995); Barbieri, RL, Am. J. Obstet.Gynecol.183: 1412-8 (2000); Kim, LH et al., Fertility and Sterility 73: 1097-1098 (2000); Iuorno, MJ et al., Obstet. Gynecol. Clin. North Am. 28: 153-164 (2001); Zacur, HZ, Obstet. Gynecol. Clin. North Am. 28: 21-33 (2001)). Recent studies suggest that androgen hypertrophy that occurs with PCOS is caused by increased ovarian androgen production (eg, testosterone and androstenedione) and decreased serum androgen-binding globulin levels due to hyperinsulinemia . Insulin has been shown to directly stimulate androgen production by the ovary by increasing the activity of P450c17α, an enzyme responsible for the production of testosterone in ovarian capsular cells (Iuorno, MJ et al., Supra). At the level of the pituitary axis, androgen hypertrophy suppresses follicle stimulating hormone (FSH) secretion, modifies gonadotropin-releasing hormone (GnRH) release, and increases luteinizing hormone (LH) secretion. Together with the local effects of androgens on the ovaries, these abnormalities lead to follicular retraction, anovulation, and infertility. Similarly, undermenstrual and amenorrhea are caused and severe vaginal bleeding occurs frequently. Hyperinsulinemia also leads to increased hypertension and thrombus formation and is considered to be associated with the development of heart disease, cerebral infarction and type 2 diabetes (Iuorno, MJ et al., Supra; Zacur, HA, supra).

  Traditionally, treatment of PCOS has been mainly directed at correcting the underlying symptoms. For example, hirsutism and menstrual irregularities have been treated with antiandrogenic drugs including oral contraceptives, spironolactone, flutamide, and finasteride. Infertility treatments included weight loss diets, ovulatory medicines (eg, clomiphene, follistim, and gonal-F), so-called “ovarian perforation” surgery, and in vitro fertilization. More recent treatments for PCOS are aimed at lowering insulin levels. Metformin, D-chiro-inositol, diazoxide, and PPAR gamma inhibitors (eg, troglitazone (Rizulin), rosiglitazone (Avandia)), And insulin sensitizers such as pioglitazone (Actos) have been shown to restore fertility and reverse PCOS-related endocrine abnormalities: metformin and PPAR gamma inhibitors Does not interfere with pregnancy, but is generally interrupted during pregnancy due to concerns about fetal safety and effects, and 30% to 50% of women with pregnant PCOS Experience a spontaneous abortion in the first three months (Iuorno, MJ et al., Supra; Zacur, HA, supra; Phipps, WR Clin. North Am. 28: 165-182 (2001), a need exists for new and better compositions and methods for treating PCOS.

SUMMARY OF THE INVENTION Applicants discover that glucagon-like peptide-1 (GLP-1), exendins, and agonists and analogs of these compounds can reduce insulin resistance or increase insulin sensitivity. The above problem was solved. The present invention relates to a method of treating PCOS using GLP-1, GLP-1 agonists, exendins, or exendin agonists. In one embodiment, the method of the invention is characterized by administering to a patient a therapeutically effective amount of a GLP-1, GLP-1 agonist, exendin, or exendin agonist. In another embodiment, the method is characterized by reducing or preventing insulin resistance in a subject afflicted with PCOS. In yet another embodiment, the method is characterized by preventing the onset of type 2 diabetes in a subject afflicted with PCOS. In a further embodiment, the method is characterized by restoring regular menstruation, ovulation, or fertility in a subject afflicted with PCOS.

  In a preferred embodiment, the GLP-1 agonist is a GLP-1 analog having agonist activity and the exendin agonist is an exendin analog having agonist activity. In another preferred embodiment, the subject is a human.

  A GLP-1 agonist means a compound that mimics the effect of GLP-1 on PCOS by binding to the receptor or receptors that cause this effect. Specific GLP-1 analogs with agonist activity are described in Chen et al., U.S. Pat.No. 5,512,549, issued Apr. 30, 1996, entitled Glucagon-Like Insulinotropic Peptide Analogs, Compositions and Methods of Use. ing. Other GLP-1 analogs with agonist activity are described in US Pat. No. 5,574,008, published in Johnson et al., November 12, 1996, entitled Biologically Active Fragments of Glucagon-Like Insulinotropic Peptides. Still other GLP-1 analogs with agonist activity are described in US Pat. No. 5,545,618 entitled GLP-1 Analogs Useful for Diabetes Treatment, issued August 13, 1996, Buckley et al. . All three referenced US patents are hereby incorporated by this reference.

  An exendin agonist means a compound that mimics the effect of exendin on PCOS by binding to the receptor or receptors that cause this effect. Certain exendin analogs with agonist activity are described in U.S. Provisional Patent Application No. 60 / 055,404, filed Aug. 8, 1997, which is in common with the present invention and incorporated herein by reference. Has been. Certain other exendin analogs with agonist activity are shared with the present invention and are incorporated herein by reference, US Provisional Patent Application No. 25 application number 60 filed November 14, 1997. / 066,029 and 60 / 065,44. Preferred exendin analogs having agonist activity include those described in US Provisional Patent Application Nos. 60 / 055,404 and 60 / 065,442.

DETAILED DESCRIPTION OF THE INVENTION Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present application, including definitions, will control. All publications, patents and other references cited herein are incorporated by reference.

  Although methods and materials similar or equivalent to those described herein are used in the practice of the present invention, suitable methods and materials are described below. The materials, methods, and examples are for illustrative purposes only and are not intended to be limiting. Other features and advantages of the invention will be apparent from the detailed description, and from the claims.

  In order to further define the invention, the following terms and definitions are provided herein.

  As used herein, the term “hair loss” refers to a condition in which a patient experiences hair loss due to a specific disease such as a scalp or skin infection, nervousness, or PCOS. The hair may fall out in some places, or instead of a full baldness in one area, it may be a loss of hair extensiveness.

  The term “exendin” refers to the naturally occurring exendin peptide found in salivary secretion of the Gila-monster. The term exendin includes exendin-3 [SEQ ID NO: 7] present during the salivary secretion of Heloderma harridum, and exendin-4 [SEQ ID NO: 9], a peptide present during the salivary secretion 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). Exendin-4, which occurs during saliva secretion in the Gila monster, is an amidated peptide. However, it is to be understood that the terms “exendin”, “exendin-3”, and “exendin-4” refer to both the amidated form of the peptide and the acid form of the peptide.

Exendin-4 was initially thought to be a (potentially toxic) poison component. Currently, exendin-4 lacks toxicity and appears to be made in the salivary glands of the Gila Monster instead. Exendin has some sequence similarity with several members of the glucagon-like peptide family, the highest homology being 53% to GLP-1 [7-36] NH ₂ (Goke et al., J. Biol Chem., 268: 19650-55, 1993). “Exendin agonist” refers to a compound that mimics the effects of exendin by binding to the receptor or receptors that cause this effect. An exendin “acting activity” in this context means having the biological activity of exendin, but the activity of an analog can be either more potent or less potent than natural exendin. Understood. Other exendin agonists include, for example, chemical compounds specifically designed to activate a receptor or receptors that have an effect on PCOS.

The terms “GLP-1” and “glucagon-like peptide-1” according to the present invention refer to the acid form of GLP-1, also known as GLP-1 amide [SEQ ID NO: 4], as well as proglucagon [78-107]. Point to. “GLP-1 agonist” refers to a compound that mimics the effect of GLP-1 on PCOS by binding to the receptor or receptors that GLP-1 causes this effect. A preferred GLP-1 agonist is a GLP-1 analog having agonist activity. GLP-1 “agonist activity” in this context means having the biological activity of GLP-1 (7-36) amide (GLP-1 (7-36) NH ₂ ), but the activity of the analog is It is understood that it can be either less potent or more potent than natural GLP-1 (7-36) amide. Other GLP-1 agonists include, for example, chemical compounds specifically designed to activate a receptor or receptors that GLP-1 affects PCOS.

  As used herein, the term “hirutism” refers to a condition in which a patient exhibits abnormal hair depth.

  As used herein, the term “androgen excess” refers to a condition in which a patient exhibits elevated levels of androgens (eg, testosterone, androstenedione) in serum.

  As used herein, the term “hyperinsulinemia” refers to a condition in which a patient exhibits elevated plasma insulin levels.

  As used herein, the term “hyperlipidemia” refers to a condition in which a patient exhibits elevated concentrations of any or all lipids in plasma.

  As used herein, the term “hypertension” means that a patient has a persistently high blood pressure (ie, a diastolic pressure equal to or greater than 140 mm Hg and greater than or equal to 90 mm Hg). Pressure).

  As used herein, the term “insulinotropic” refers to the ability to stimulate the release of insulin into the circulation.

  As used herein, the term “insulin resistance” refers to a subnormal biological response to a given concentration of insulin (ie, a decrease in glucose transport across the cell membrane in response to insulin). .

  As used herein, the term “pharmaceutically acceptable carrier or adjuvant” may be administered to a patient together with a compound of the present invention and does not destroy its pharmacological activity. Or refers to an adjuvant.

  As used herein, the terms “polycystic ovary syndrome”, “PCOS”, “polycystic ovary” or “Stein-Lebental syndrome” refer to a disease that affects women. A woman diagnosed with PCOS may have one or more of the following symptoms: anovulation (irregular or amenorrhea), hyperandrogenism (elevated serum testosterone and / or androstenedione), abnormal Uterine bleeding, enlarged polyfollicular ovary, infertility, obesity, insulin resistance, hyperinsulinemia, hypertension, hyperlipidemia, type 2 diabetes, excessive facial hair growth, hair loss, and acne.

  The term “therapeutically or pharmaceutically effective” or “therapeutically or pharmaceutically effective amount” refers to PCOS or its symptoms (eg, insulin resistance for some period of time, hyperinsulinemia, type 2 diabetes, obesity, hypertension, high Reduce or alleviate the severity of any of (lipidemia, anovulation or irregular ovulation, infertility, hyperandrogenism, hirsutism, hair loss, acne, enlarged polyfollicular ovary and abnormal uterine bleeding) Refers to the amount of the compound of the invention required to Also, a therapeutically or pharmaceutically effective amount means an amount necessary to improve a patient's clinical symptoms.

  As used herein, the term “type 2 diabetes” is also known as non-insulin dependent diabetes mellitus (NIDDM) or adult-onset diabetes (AODM) in which the patient has an elevated concentration of blood glucose levels. Refers to illness.

  The present invention relates to a method of treating PCOS in a patient. The method includes administering to the subject a therapeutically effective amount of GLP-1, exendin, and agonists and analogs of these compounds. Furthermore, the methods of the invention relate to reducing insulin resistance using GLP-1, exendins, and agonists and analogs of these compounds. Many of the symptoms associated with PCOS are due to underlying insulin resistance.

Glucagon-like peptide-1 (GLP-1)
GLP-1 plays an important role in the control of plasma glucose homeostasis. It is involved in stimulating insulin secretion, inhibiting glucagon release by the pancreas, inhibiting gastric acid secretion and motility, and suppressing appetite and food intake. GLP-1 is a member of the incretin group of secretagogue hormones released from enteroendocrine cells in response to food intake. GLP-1 binds to the GLP-1 receptor, which is expressed on pancreatic β cells. Binding of GLP-1 to its receptor triggers an intracellular signaling pathway that, in conjunction with inhibition of glucagon production, results in stimulation of insulin secretion. This in turn leads to inhibition of glucose production in the liver which lowers blood glucose levels. Although the role of GLP-1 in maintaining blood glucose levels was established prior to the present invention, it was not known that GLP-1 could increase insulin sensitivity.

Mammalian GLP peptides and glucagon are encoded by the same gene. In the ileum, precursors are processed into two main classes of GLP peptide hormones, GLP-1 and GLP-2. GLP-1 (1-37) has the sequence: His-Asp-Glu-Phe-Glu-Arg-His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr- It has Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-Gly [SEQ ID NO: 1]. GLP-1 (1-37) is post-translationally amidated and has the sequence: His-Asp-Glu-Phe-Glu-Arg-His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp -Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg (NH ₂ ) [SEQ ID NO: or obtaining a GLP-1 (1-36) NH ₂ having a 2 ', or is enzymatically processed sequence: His-Ala-Glu-Gly -Thr-Phe-Thr-Ser-Asp-Val-Ser GLP- with -Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-Gly [SEQ ID NO: 3] Obtain 1 (7-37). In addition, GLP-1 (7-37) is amidated and has the sequence: His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly GLP-1 (7-36) amide having -Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg (NH ₂ ) [SEQ ID NO: 4] May be obtained. Similarly, GLP-1 (1-36) amide is processed to GLP-1 (7-36) amide.

Intestinal L cells secrete GLP-1 (7-37) and GLP-1 (7-36) NH ₂ at a ratio of about 1: 5. These incomplete forms of GLP-1 have a short half-life in vivo (less than 10 minutes), which are inactivated by aminodipeptidase (DPP IV), each with the sequence: Glu-Gly-Thr -Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg GLP-1 (9-37) with -Gly [SEQ ID NO: 5] and the sequence: Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly- Obtain GLP-1 (9-36) amide with Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg (NH ₂ ) [SEQ ID NO: 6] . It has been speculated that the peptides GLP-1 (9-37) and GLP-1 (9-36) amide may have an effect on hepatic glucose production, but obviously they are from the pancreas Does not stimulate the production or release of insulin.

As used herein, the term “GLP-1 analog” refers to GLP-1 (1-37), GLP-1 (1-36) NH ₂ , GLP-1 (7-37), GLP- 1 (9-37), GLP-1 (9-36) NH ₂ (“GLP-1 (9-36) amide”). The invention includes the use of recombinant human GLP-1 analogs and GLP-1 analogs derived from other species, whether recombinant or synthetic.

  As used herein, the term “GLP-1 agonist” refers to GLP-1 (in which GLP-1 acts on PCOS, regardless of peptide, peptidomimetic, or other chemical compound). 7-36) Any molecule that binds to or activates the GLP-1 receptor or receptors, such as the amide receptor and its second messenger cascade. For example, GLP-1 agonists have insulinotropic activity and include, inter alia, molecules that are agonists of (ie, activate) GLP-1 receptor molecules and their second messenger activity against insulin-producing β-cells.

  A “GLP-1 analog” also includes a peptide encoded by a polynucleotide that expresses a biologically active GLP-1 analog having agonist activity, as defined herein. For example, a GLP-1 analog may be a peptide containing one or more amino acid substitutions, additions or deletions compared to GLP-1 (7-36) amide. In one embodiment, 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 Any integer less than or between these quantities. In one embodiment of the invention, the substitution comprises one or more conservative substitutions. A “conservative” substitution refers to the replacement of an amino acid residue with another biologically active similar residue. Examples of conservative substitutions are substitution of one hydrophobic residue such as isoleucine, valine, leucine, or methionine, or arginine to lysine, glutamic acid to aspartic acid, glutamine to asparagine Substitution of one polar residue with the other, such as The following table lists descriptive but non-limiting conservative amino acid substitutions.

  In addition, GLP-1 analogs have the above peptides chemically derivatized or modified, eg, unnatural amino acid residues (eg, taurine, β- and γ-amino acid residues and D-amino acid residues) Cyclization with C-terminal functional modifications such as amide, ester, and C-terminal ketone modifications, and N-terminal functional modifications such as acylated amines, Schiff bases, or found for example with the amino acid pyroglutamic acid It is understood to include peptides having

(1) for SEQ ID NO: 1, 2, 3, and 4; and (2) a peptide having 50% or more sequence identity, preferably 90% or more sequence identity to its incomplete sequence Sequences are also included in the present invention. As used herein, sequence identity refers to a comparison made between two molecules using standard algorithms well known in the art. A preferred algorithm for calculating sequence identity for the present invention uses SEQ ID NO: 1 [ie GLP-1 (1-37)] as a reference sequence to define the percentage identity of homologs over its length Smith-Waterman algorithm. Although some parameter values have been found to give more biologically realistic results than others, the choice of parameter values for matches, mismatches, and insertions or deletions is arbitrary. One preferred set of parameter values for the Smith-Waterman algorithm is 1 for matched residues and -1 / for mismatched residues (residues are either single nucleotides or single amino acids). It is described in the “maximum similarity segment” approach using a value of 3. Waterman, Bull. Math. Biol. 46; 473 (1984). Insertions and deletions (indels), x is x _k = 1 + 1 / 3k, where k is the residue in a given insert or deletion. Is a number of]. Ibid.

によって与えられるパーセント同一性を有するだろう。 For example, except for 18 amino acid substitutions and 3 amino acid insertions, a sequence that is identical to the 37-amino acid residue sequence of SEQ ID NO: 1 is:

Will have the percent identity given by.

Glucagon-like peptide agonists that exhibit activity via the GLP-1 (7-36) amide receptor have been described. EP 0708179 A2; Hjorth et al., J. Biol. Chem. 269; 30121 (1994); Siegel et al., Amer. Diabetes Assoc. 57th Scientific Session, Boston (1997); Hareter et al., Amer. Diabetes Assoc 57th Scientific Session, Boston (1997); Adelhorst et al., J. Biol. Chem. 269, 6275 (1994); Deacon et al., 16th International Diabetes Federation Congress Abstracts, Diabetologia Supplement (1997); Irwin et al. , Proc. Natl. Acad. Sci. USA 94; 7915 (1997); Mojsov, Int. J. Peptide Protein Res. 40; 333 (1992). Goke & Byrne, Diabetic Medicine 13; 854 (1996). Recent publications disclose black widow spider GLP-1 and Ser ² GLP-1. See Holz & Hakner, Comp. Biochem. Physiol., Part B 121; 177 (1998) and Ritzel et al., J. Endocrinol 159; 93 (1998).

  The GLP-1 receptor is a cell surface protein found, for example, on insulin-producing pancreatic β-cells; the GLP-1 (7-36) receptor has been characterized in the art. Methods of determining whether a chemical or peptide binds to or activates the GLP-1 receptor are known to those skilled in the art.

  The biological activity of GLP-1 agonists can be determined by in vitro and in vivo animal models and human studies well known to those skilled in the art. GLP-1 biological activity is determined by standard methods, generally suitable cells that express the GLP-1 receptor on their surface, for example insulinoma cell lines such as RINmSF cells or INS-1 cells. Can be determined by a receptor binding activity screening procedure. See Mojsov, Int. J. Peptide Protein Res. 40; 333 (1992) and EP 0708179 A2. Alternatively, cells created to express the GLP-1 receptor can be used. In addition to measuring specific binding of the tracer to the membrane using radioimmunoassay methods, cAMP activity or glucose dependent insulin production can also be measured. In one method, a polynucleotide encoding a GLP-1 receptor is used to transfect the cell so that the cell expresses the GLP-1 receptor protein. Thus, for example, these methods contact such cells with a compound to be screened and determine whether such a compound generates a signal (ie, activates a receptor), It can be used in screening for receptor agonists. Other screening techniques include the use of cells that express the GLP-1 receptor, such as transfected CHO cells, in systems that measure extracellular pH or ionic changes caused by receptor activation. For example, a potential agonist may be contacted with a cell that expresses a GLP-1 protein receptor, and a second messenger response (eg, signal transduction or ion or pH change) is measured to determine the potential agonist. Can be measured.

  Polyclonal and monoclonal antibodies can be used to detect, purify, and identify GLP-1-like peptides for use in the methods described herein. Antibodies such as ABGA1178 detect intact GLP-1 (1-37) or N-terminally truncated GLP-1 (7-37) or GLP-1 (7-36) amide. Other antibodies detect the C-terminal end of the precursor molecule, and the procedure is subtracted by biologically active truncated peptide (i.e. GLP-1 (7-37) amide) Makes it possible to calculate the amount of. Orskov et al., Diabetes 42; 658 (1993); Orskov et al., J. Clin. Invest. 1991, 87; 415 (1991).

  The GLP-1 and GLP-1 agonists of the present invention are peptides that can be made by solid state chemical peptide synthesis. Such peptides can also be made by conventional recombinant techniques using standard procedures as described, for example, in Sambrook & Maniatis, Molecular Cloning, A Laboratory Manual. As used herein, “recombinant” is a set in which a gene has been genetically modified to contain a polynucleotide encoding a GLP-1 peptide, as described herein. It is derived from the alternative (microbe or mammal) expression system.

  The GLP-1 and GLP-1 agonists of the present invention are naturally purified products, or products of synthetic chemical procedures, or prokaryotic or eukaryotic hosts (eg, bacteria, yeast, higher in culture or in vivo) A peptide produced by recombinant technology from plants, insects or mammalian cells). Depending on the host used in the recombinant production procedure, the polypeptides of the invention may generally be non-glycosylated or glycosylated. GLP-1 peptides include ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography, and lectin chromatography. , And can be recovered and purified from the recombinant cell culture by methods not limited to these. High performance liquid chromatography (HPLC) can be used for final purification steps.

Exendin Exendin-4, 39-amino acid polypeptide is a naturally occurring peptide isolated from the salivary secretion of Gila Monster. Animal studies with exendin-4 show that its ability to lower blood glucose persists for several hours. Exendin-4, which occurs in the saliva secretion of Gila Monster, is an amidated peptide. However, it is to be understood that the terms “exendin” and “exendin-4” specifically refer to both the amidated form of the peptide and the acid form of the peptide.

  Specific exendin sequences are compared in Table 1 to the sequence of GLP-1.

  As described herein, the nonclinical pharmacology of exendin-4 has been studied. In the brain, exendin-4 binds mainly to the last area of the hindbrain and the solitary nucleus region and to the frontal subfornical organ. Exendin-4 binding has been observed in rat and mouse brain and kidney. The structure of exendin-4 binding in the kidney is unknown.

Various experiments compare the biological effects of exendin-4 and GLP-1 and show a more favorable spectrum of properties for exendin-4. A single subcutaneous dose of exendin-4 reduced plasma glucose in db / db (diabetic) and ob / ob (diabetic and obese) mice by up to 40%. In Diabetic Fatty Zucker (ZDF) rats, 5 weeks of treatment with exendin-4 reduced HbA _1c (measurement of glycosylated hemoglobin used to assess blood glucose levels) by up to 41%. Insulin sensitivity was also improved by 76% in obese ZDF rats after 5 weeks of treatment. A dose-dependent decrease in plasma glucose was also observed in glucose intolerant primates.

  Also, the insulinotropic effect of exendin-4 was observed in rodents, more than 100% in unfasted Harlan Sprague Dawley (HSD) rats, up to 10 times in unfasted db / db mice Improved insulin response to glucose. Higher pretreatment plasma glucose concentrations were associated with greater glucose lowering effects. That is, the observed glucose-lowering effect of exendin-4 appears to be glucose dependent, minimal if the animal is already normoglycemic.

  Exendin-4 dose slowed gastric emptying in a dose-dependent manner in HSD rats and was -90 times more potent than GLP-1 for this effect. Exendin-4 was also shown to reduce food intake in NIH / Sw (Swiss) mice following terminal administration, which was at least 1000 times more potent than GLP-1 for this effect . Exendin-4 decreased plasma glucagon levels by approximately 40% in anesthetized ZDF rats during hyperinsulinic and hyperglycemic fixation, but plasma glucagon in normal rats during normoglycemia Concentration was not affected. Exendin-4 has been shown to reduce body weight in a dose-dependent manner in obese ZDF rats, but in lean ZDF rats, the observed decrease in body weight appears to be transient.

  The toxicology of exendin-4 has been investigated in single dose studies in mice, rats and monkeys, repeated dose (up to 28 consecutive daily doses) studies in rats and monkeys and in vitro tests of mutagenicity and chromosomal alterations . To date, no deaths have occurred, and no treatment-related changes have been found in hematology, clinical chemistry, or global or microscopic tissue changes. Exendin-4 was shown to be non-mutagenic and did not cause chromosomal abnormalities at the concentrations tested (up to 5000 μg / mL).

In addition to investigating nonclinical pharmacokinetics and metabolism of exendin-4, a number of immunoassays have been developed. A radioimmunoassay with limited sensitivity (˜100 pM) was used in the initial pharmacokinetic study. Subsequently, a two-site IRMA assay for exendin-4 was confirmed with a lower quantification limit of 15 pM. The bioavailability of exendin-4 given subcutaneously was found to be about 50-80% using radioimmunoassay. This was similar to that seen after intraperitoneal administration (48-60%). Peak plasma concentrations (C _max ) occurred between 30 and 43 minutes (T _max ). Both C _max and AUC values were monotonically related to dose. The apparent final half-life of exendin-4 given subcutaneously was approximately 90-110 minutes. This was significantly longer than the 14-41 minutes seen after the intravenous dose. Similar results were obtained using the IRMA assay. The degradation test of exendin-4 compared to GLP-1 shows that exendin-4 is relatively resistant to degradation.

  As used herein, the term “exendin agonist” refers to the effect of exendin on PCOS, such as the GLP-1 (7-36) amide receptor, and its second messenger cascade. Any molecule is included, whether it is a peptide, peptidomimetic, or other chemical compound that binds to or activates a receptor or receptors at a position that exerts. For example, exendin agonists include molecules that have insulinotropic activity and GLP-1 receptor molecules and especially those that are agonists (ie, activate) of their second messenger activity against insulin-producing β cells.

  Exendin structure-activity relationship (SAR) has been investigated for structures that may be related to exendin's activity in terms of stability to metabolism and improvement of physical characteristics, in particular it is peptide stability and submission to alternative delivery systems. Therefore, various exendin agonist peptide compounds have been invented. Exendin agonists include exendin analogs having agonist activity in which one or more naturally occurring amino acids are inserted, eliminated or replaced by another amino acid (s). A preferred exendin analog is a peptide analog of exendin-4.

  Exendin analogs include peptides encoded by a polynucleotide that expresses a biologically active exendin analog having agonist activity, as defined herein. For example, an exendin analog may be a peptide containing one or more amino acid substitutions, additions or deletions compared to exendin-4. In one embodiment, 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 Any integer less than or between these quantities. In one embodiment of the invention, the substitution comprises one or more conservative substitutions. A “conservative” substitution refers to the replacement of an amino acid residue with another biologically active similar residue. Examples of conservative substitutions are substitution of one hydrophobic residue such as isoleucine, valine, leucine, or methionine, or arginine to lysine, glutamic acid to aspartic acid, glutamine to asparagine Substitution of one polar residue with the other, such as The following table lists descriptive but non-limiting conservative amino acid substitutions.

さらに、エキセンディンアナログは、化学的に誘導体化または改変された上記のペプチド、例えば、非天然アミノ酸残基(例えば、タウリン、β-およびγ-アミノ酸残基およびD-アミノ酸残基)を有する、アミド、エステル、およびC-末端ケトン修飾のようなC-末端官能基修飾、およびアシル化アミン、シッフ塩基のようなN-末端官能基修飾を有する、または例えばアミノ酸ピログルタミン酸で見出される環化を有するペプチドを含むと理解される。

In addition, exendin analogs have the above peptides chemically derivatized or modified, for example, unnatural amino acid residues (e.g., taurine, β- and γ-amino acid residues and D-amino acid residues), Cyclization with C-terminal functional group modifications such as amide, ester, and C-terminal ketone modifications, and N-terminal functional group modifications such as acylated amines, Schiff bases, or found for example with the amino acid pyroglutamic acid It is understood to include peptides having.

Peptide sequences having (1) SEQ ID NOs: 7 and 9; and (2) 50% or more sequence identity, preferably 90% or more sequence identity to the incomplete sequence include. As used herein, sequence identity refers to a comparison made between two molecules using standard algorithms well known in the art. A preferred algorithm for calculating sequence identity for the present invention is the Smith-Waterman algorithm that uses SEQ ID NO: 9 [ie, exendin-4] as a reference sequence to define the percentage identity of homologs over its length. It is. Although some parameter values have been found to give more biologically realistic results than others, the choice of parameter values for matches, mismatches, and insertions or deletions is arbitrary. One preferred set of parameter values for the Smith-Waterman algorithm is 1 for matched residues and -1 / for mismatched residues (residues are either single nucleotides or single amino acids). It is described in the “maximum similarity segment” approach using a value of 3. Waterman, Bull. Math. Biol. 46; 473 (1984). Insertions and deletions (indels), x is x _k = 1 + 1 / 3k, where k is the residue in a given insert or deletion. Is a number of]. Ibid.

  A novel exendin analog with agonist activity is disclosed in "Novel Exendin Agonist" filed on August 6, 1998, claiming the benefit of US Patent Application No. 60 / 055,404, filed August 8, 1997. Commonly owned PCT application number PCT / US98 / 16387 entitled “Compounds”, both of which are incorporated herein by reference.

  Other novel exendin analogs with agonist activity were identified in November 1998 entitled “Novel Exendin Agonist Compounds” claiming the benefit of US Provisional Application No. 60 / 065,442, filed November 14, 1997. Commonly-owned PCT application No. PCT / US98 / 24210 filed on the 13th, both of which are incorporated herein.

  Still other novel exendin analogs with agonist activity are listed in November 1998 entitled “Novel Exendin Agonist Compounds” claiming the benefit of US Provisional Application No. 60 / 066,029, filed November 14, 1997. Commonly-owned PCT application number PCT / US98 / 24273 filed on the 13th of the month, both of which are incorporated herein by reference.

  Yet another exendin analog having agonist activity was entitled “Methods for Regulating Gastrointestinal Activity”, a continuation-in-part of US patent application Ser. No. 08 / 694,954 filed Aug. 8, 1996. No. PCT / US97 / 14199 filed Aug. 8, 1997, commonly owned PCT Application No. PCT / US97 / 14199, both of which are incorporated herein by reference.

  Still other exendin analogs with agonist activity are claimed in US Provisional Application No. 60 / 034,90 filed Jan. 7, 1997, "Use of Exendins and Agonist Thereof for The commonly owned PCT application number PCT / US98 / 00449, filed January 7, 1998, entitled “the Reduction of Food Intake”, both of which are incorporated herein by reference. Incorporated inside.

  Still other exendin analogs with agonist activity are claimed in US Provisional Application No. 60 / 175,365, filed January 10, 2000, "Use of Exendins and Agonist Thereof for Modulation of The commonly owned PCT application number PCT / US01 / 00719, filed Jan. 9, 2001, entitled “Triglyceride Levels and Treatment of Dyslipidemia”, both of which are described herein. Incorporated inside.

  Still other exendin analogs with agonist activity are claimed in US Provisional Application No. 60 / 116,380, filed January 14, 1999, "Novel Exendin Agonist Formulations and Methods of Administration Thereof No. PCT / US00 / 00902, filed Jan. 14, 2000, entitled “", both of which are incorporated herein by reference.

  Still other exendin analogs with agonist activity are claimed as "Novel Exendin Agonist Formulations and Methods of Administration Thereof", claiming priority to US Application No. 10 / 157,224, filed May 28, 2002. In commonly owned PCT application number PCT / US03 / 16699, filed May 28, 2003, both of which are incorporated herein by reference.

  Yet another exendin analog with agonist activity was entitled "Methods of Glucagon Suppression", claiming priority to US Provisional Application No. 60 / 132,017, filed April 30, 1999 No. PCT / US00 / 00942 filed Jan. 14, 2000, commonly owned PCT application number PCT / US00 / 00942, both of which are incorporated herein by reference.

  Still other exendin analogs with agonist activity are claimed in US Application No. 09 / 323,867, filed June 1, 1999, "Use of Exendins and Agonist Thereof for the Treatment of Gestational Diabetes Mellitus "is listed in commonly owned PCT Application No. PCT / US00 / 14231 filed May 23, 2000, both of which are incorporated herein by reference. Is done.

  Yet another exendin analog having agonist activity claims priority to US Provisional Application No. 60 / 075,122, filed February 13, 1998, “Inotropic and Diuretic Effects of Exendin and GLP-1 In commonly owned PCT Application No. PCT / US99 / 02554, filed February 5, 1999, both of which are incorporated herein by reference.

  The activity as exendin agonists and exendin analogs having agonist activity can be shown, for example, by activity in the following assay. The effects of multiple exendins or exendin agonists on PCOS can be identified, evaluated, or using methods described herein, or methods for determining effects on PCOS known or equivalent in other fields, or Can be screened. Screening for exendin agonist compounds or candidate exendin agonist compounds such as negative receptor assay or GLP-1 receptor assay / screening above, amylin receptor assay / content incorporated herein by reference 1993 Screening with amylin receptor preparations, as described in US Pat. No. 5,264,372 issued on Jan. 23, one or more calcitonin receptor assays / eg calcium reception coupled to stimulation of adenyl cyclase activity Screening with T47D and MCF7 breast cancer cells containing the body, and / or CGRP receptor assay / eg screening with SK-N-MC cells.

Certain preferred exendin analogs with agonist activity are:
Exendin-4 (1-30) [SEQ ID NO: 14: His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly];
Exendin-4 (1-30) amide [SEQ ID NO: 15: His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly-NH ₂ ];
Exendin-4 (1-28) amide [SEQ ID NO: 16: His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn-NH ₂ ];
¹⁴ Leu, ²⁵ Phe exendin-4 amide [SEQ ID NO: 17: His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser-NH ₂ ];
¹⁴ Leu, ²⁵ Phe exendin-4 (1-28) amide [SEQ ID NO: 18: His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn- NH ₂ ]; and
¹⁴ Leu, ²² Ala, ²⁵ Phe exendin-4 (1-28) amide [SEQ ID NO: 19: His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu Glu Ala Val Arg Leu Ala Ile Glu Phe Leu Lys Asn-NH ₂ ].

  Also included within the scope of the invention are pharmaceutically acceptable salts of the compounds of formula (I-VIII) and pharmaceutical compositions comprising the compounds and salts thereof.

Formula I
An exendin analog having agonist activity is represented by the formula (I) [SEQ ID NO: 20]:
Xaa ₁ Xaa ₂ Xaa ₃ Gly Xaa ₅ Xaa ₆ Xaa ₇ Xaa ₈ Xaa ₉ Xaa ₁₀
Xaa ₁₁ Xaa ₁₂ Xaa ₁₃ Xaa ₁₄ Xaa ₁₅ Xaa ₁₆ Xaa ₁₇ Ala Xaa ₁₉ Xaa ₂₀
Xaa ₂₁ Xaa ₂₂ Xaa ₂₃ Xaa ₂₄ Xaa ₂₅ Xaa ₂₆ Xaa ₂₇ Xaa ₂₈ -Z ₁ ;
[Where:
Xaa ₁ is His, Arg or Tyr;
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;
Xaa ₁₀ is Ala, Leu, Ile, Val, pentylglycine or Met;
Xaa ₁₁ is Ala or Ser;
Xaa ₁₂ is Ala or Lys;
Xaa ₁₃ is Ala or Gln;
Xaa ₁₄ is Ala, Leu, Ile, pentylglycine, Val or Met;
Xaa ₁₅ is Ala or Glu;
Xaa ₁₆ is Ala or Glu;
Xaa ₁₇ is Ala or Glu;
Xaa ₁₉ is Ala or Val;
Xaa ₂₀ is Ala or Arg;
Xaa ₂₁ is Ala or Leu;
Xaa ₂₂ is Ala, Phe, Tyr or naphthylalanine;
Xaa ₂₃ is Ile, 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;
Xaa ₂₇ is Ala or Lys;
Xaa ₂₈ is Ala or Asn;
Z ₁ is -OH,
-NH ₂
Gly-Z ₂ ,
Gly Gly-Z ₂ ,
Gly Gly Xaa ₃₁ -Z ₂ ,
Gly Gly Xaa ₃₁ Ser-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ -Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ -Z ₂ or
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ Xaa ₃₈ -Z ₂ ;
Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ 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 ;
_{However, Xaa 3, Xaa 5, Xaa} 6, Xaa 8, Xaa 10, Xaa 11, Xaa 12, Xaa 13, Xaa 14, Xaa 15, Xaa 16, Xaa 17, Xaa 19, Xaa 20, Xaa 21, Xaa 24, Xaa ₂₅ , Xaa ₂₆ , Xaa ₂₇ and Xaa ₂₈ are Ala.], And those described in US Provisional Application No. 60 / 065,442.

  Preferred N-alkyl groups for N-alkylglycine, N-alkylpentylglycine and N-alkylalanine preferably comprise a lower alkyl group of 1 to about 6 carbon atoms, more preferably 1 to 4 carbon atoms.

Preferred exendin analogs include those where Xaa ₁ is His or Tyr. More preferably, Xaa ₁ is His.

A compound in which Xaa ₂ is Gly is preferred.

Compounds in which Xaa ₁₄ is Leu, pentylglycine or Met are preferred.

Preferred compounds are those where Xaa ₂₅ is Trp or Phe.

Preferred compounds are those in which Xaa ₆ is Phe or naphthylalanine; Xaa ₂₂ is Phe or naphthylalanine and Xaa ₂₃ is Ile or Val.

Preference is given to compounds in which Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ and Xaa ₃₈ are independently selected from Pro, homoproline, thioproline and N-alkylalanine.

Preferably Z ₁ is —NH ₂ .

Preferably Z ₂ is —NH ₂ .

According to one embodiment, formula (I) [wherein Xaa ₁ is His or Tyr, more preferably His; Xaa ₂ is Gly; Xaa ₆ is Phe or naphthylalanine; Xaa ₁₄ is Leu, pentylglycine or Met; Xaa ₂₂ is Phe or naphthylalanine; Xaa ₂₃ is Ile or Val; Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ and Xaa ₃₈ are independently , Pro, homoproline, thioproline or N-alkylalanine]. More preferably, Z ₁ is —NH ₂ .

According to a particularly preferred embodiment, a particularly preferred compound is of the formula (I) [wherein Xaa ₁ 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 neftylalanine; Xaa ₇ is Thr or Ser; Xaa ₈ is Ala, Ser or Thr; Xaa ₉ is Asp Xaa ₁₀ is Ala, Leu or pentylglycine; Xaa ₁₁ is Ala or Ser; Xaa ₁₂ is Ala or Lys; Xaa ₁₃ is Ala or Gln; Xaa ₁₄ Is Ala, Leu or pentylglycine; Xaa ₁₅ is Ala or Glu; Xaa ₁₆ is Ala or Glu; Xaa ₁₇ is Ala or Glu; Xaa ₁₉ is Ala or Val Xaa ₂₀ is Ala or Arg; Xaa ₂₁ is Ala or Leu; Xaa ₂₂ is Phe or naphthylalanine; Xaa ₂₃ is Ile, Val or tert-butylglycol. 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; Z ₁ is -OH, -NH ₂ , Gly-Z ₂ , Gly Gly-Z ₂ , Gly Gly Xaa ₃₁ -Z ₂ , Gly Gly Xaa ₃₁ Ser-Z ₂ , Gly Gly Xaa ₃₁ Ser Ser-Z ₂ , Gly Gly Xaa ₃₁ Ser Ser Gly-Z ₂ , Gly Gly Xaa ₃₁ Ser Ser Gly Ala-Z ₂ , Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ -Z ₂ , Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ -Z ₂ , Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ Xaa ₃₈ -Z ₂ ; Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ and Xaa ₃₈ are independently Pro homoproline, Thioproline or N-methylalanine; and Z ₂ is —OH or —NH ₂ ; provided that Xaa ₃ , Xaa ₅ , Xaa ₆ , Xaa ₈ , Xaa ₁₀ , Xaa ₁₁ , Xaa ₁₂ , Xaa ₁₃ , Xaa ₁₄ , Xaa ₁₅ , Xaa ₁₆ , Xaa ₁₇ , Xaa ₁₉ , Xaa ₂₀ , Xaa ₂₁ , Xaa ₂₄ , Xaa ₂₅ , Xaa ₂₆ , Xaa ₂₇ and Xaa ₂₈ Less than one is Ala]. A particularly preferred compound is described in PCT Application No. PCT / US98 / 24210 filed November 13, 1998 entitled "Novel Exendin Agonist Compounds", among which identified as Compound 2-23 Including things.

According to a particularly preferred embodiment, Xaa ₁₄ is Leu, Ile, Val or pentylglycine, more preferably Leu or pentylglycine, and Xaa ₂₅ is Phe, Tyr or naphthylalanine, more preferably Phe or naphthylalanine. A compound is provided. These compounds will be less sensitive to oxidative degradation reactions both in vitro and in vivo, and during compound synthesis.

Formula II
An exendin analog having agonist activity is represented by the formula (II) [SEQ ID NO: 21]:
Xaa ₁ Xaa ₂ Xaa ₃ Xaa ₄ Xaa ₅ Xaa ₆ Xaa ₇ Xaa ₈ Xaa ₉ Xaa ₁₀
Xaa ₁₁ Xaa ₁₂ Xaa ₁₃ Xaa ₁₄ Xaa ₁₅ Xaa ₁₆ Xaa ₁₇ Ala Xaa ₁₉ Xaa ₂₀
Xaa ₂₁ Xaa ₂₂ Xaa ₂₃ Xaa ₂₄ Xaa ₂₅ Xaa ₂₆ Xaa ₂₇ Xaa ₂₈ -Z ₁ ;
[Where:
Xaa ₁ 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 ₄ 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;
Xaa ₁₀ is Ala, Leu, Ile, Val, pentylglycine or Met;
Xaa ₁₁ is Ala or Ser;
Xaa ₁₂ is Ala or Lys;
Xaa ₁₃ is Ala or Gln;
Xaa ₁₄ is Ala, Leu, Ile, pentylglycine, Val or Met;
Xaa ₁₅ is Ala or Glu;
Xaa ₁₆ is Ala or Glu;
Xaa ₁₇ is Ala or Glu;
Xaa ₁₉ is Ala or Val;
Xaa ₂₀ is Ala or Arg;
Xaa ₂₁ is Ala or Leu;
Xaa ₂₂ is Phe, Tyr or naphthylalanine;
Xaa ₂₃ is Ile, 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;
Xaa ₂₇ is Ala or Lys;
Xaa ₂₈ is Ala or Asn;
Z ₁ is -OH,
-NH ₂ ,
Gly-Z ₂ ,
Gly Gly-Z ₂ ,
Gly Gly Xaa ₃₁ -Z ₂ ,
Gly Gly Xaa ₃₁ Ser-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ -Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ -Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ Xaa ₃₈ -Z ₂ or
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ Xaa ₃₈ Xaa ₃₉ -Z ₂ ;
Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ and Xaa ₃₈ are independently selected from Pro, homoproline, 3Hyp, 4Hyp, thioproline, N-alkylglycine, N-alkylpentylglycine or N-alkylalanine; and
Z ₂ is -OH or -NH ₂ ;
However, Xaa ₃ , Xaa ₄ , Xaa ₅ , Xaa ₆ , Xaa ₈ , Xaa ₉ , Xaa ₁₀ , Xaa ₁₁ , Xaa ₁₂ , Xaa ₁₃ , Xaa ₁₄ , Xaa ₁₅ , Xaa ₁₆ , Xaa ₁₇ , Xaa ₁₉ , Xaa ₂₀ , Within 3 of Xaa ₂₁ , Xaa ₂₄ , Xaa ₂₅ , Xaa ₂₆ , Xaa ₂₇ and Xaa ₂₈ are Ala; provided that Xaa ₁ is His, Arg or Tyr, then Xaa ₃ , Xaa ₄ and at least one of Xaa ₉ is Ala], including those described in US Provisional Application No. 60 / 066,029.

  Preferred N-alkyl groups for N-alkylglycine, N-alkylpentylglycine and N-alkylalanine preferably include lower alkyl groups having 1 to about 6 carbon atoms, more preferably 1 to 4 carbon atoms. Suitable compounds of the formula (II) are identified in Example 1-89 (respectively “Compound 1-89”) and filed on November 13, 1998 entitled “Novel Exendin Agonist Compounds” No. PCT / US98 / 24273 and the corresponding compounds identified in Examples 104 and 105.

Preferred such exendin analogs include those where Xaa ₁ is His, Ala or Norval. More preferably, Xaa ₁ is His or Ala. Most preferably Xaa ₁ is His.

Compounds of formula (II) [wherein Xaa ₂ is Gly] are preferred.

Compounds of formula (II) [wherein Xaa ₃ is Ala] are preferred.

Preference is given to compounds of the formula (II) [wherein Xaa ₄ is Ala].

Compounds of formula (II) [wherein Xaa ₉ is Ala] are preferred.

Compounds of formula (II) [wherein Xaa ₁₄ is Leu, pentylglycine or Met] are preferred.

Preferred compounds of formula (II) are those wherein Xaa ₂₅ is Trp or Phe.

Preferred compounds of formula (II) are those wherein Xaa ₆ is Ala, Phe or naphthylalanine; Xaa ₂₂ is Phe or naphthylalanine; and Xaa ₂₃ is Ile or Val.

Preference is given to compounds of the formula (II) in which Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ and Xaa ₃₈ are independently selected from Pro, homoproline, thioproline and N-alkylalanine.

Preferably Z ₁ is —NH ₂ .

Preferably Z ₂ is —NH ₂ .

According to one embodiment, formula (II) wherein Xaa ₁ is Ala, His or Tyr, more preferably Ala or His; Xaa ₂ is Ala or Gly; Xaa ₆ is Phe or naphthylalanine Xaa ₁₄ is Ala, Leu, pentylglycine or Met; Xaa ₂₂ is Phe or naphthylalanine; Xaa ₂₃ is Ile or Val; Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ and Xaa ₃₈ are independently , Pro, homoproline, thioproline or N-alkylalanine; and Xaa ₃₉ is Ser or Tyr, more preferably Ser]. More preferably, Z ₁ is —NH ₂ .

According to a particularly preferred embodiment, a particularly preferred compound is of the formula (II) wherein Xaa ₁ 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 Xaa ₁₀ is Ala, Leu or pentylglycine; Xaa ₁₁ is Ala or Ser; Xaa ₁₂ is Ala or Lys; Xaa ₁₃ is Ala or Gln; Xaa ₁₄ is Ala Xaa ₁₅ is Ala or Glu; Xaa ₁₆ is Ala or Glu; Xaa ₁₇ is Ala or Glu; Xaa ₁₉ is Ala or Val; Xaa ₂₀ is Ala Or Arg; Xaa ₂₁ is Ala or Leu; Xaa ₂₂ is Phe or naphthylalanine; Xaa ₂₃ is Ile, Val or tert-butylglycine; X aa 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; Z ₁ is -OH, -NH ₂ , Gly-Z ₂ , Gly Gly-Z ₂ , Gly Gly Xaa ₃₁ -Z ₂ , Gly Gly Xaa ₃₁ Ser-Z ₂ , Gly Gly Xaa ₃₁ Ser Ser-Z ₂ , Gly Gly Xaa ₃₁ Ser Ser Gly-Z ₂ , Gly Gly Xaa ₃₁ Ser Ser Gly Ala-Z ₂ , Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ -Z ₂ , Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ -Z ₂ , Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ Xaa ₃₈ -Z ₂ or Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ Xaa ₃₈ Xaa ₃₉ -Z ₂ ; Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ and Xaa ₃₈ is independently Pro homoproline, thioproline or N-methylalanine; and Z ₂ is —OH or —NH ₂ ; provided that Xaa ₃ , Xaa ₅ , Xaa ₆ , Xaa ₈ , Xaa ₁₀ , Xaa ₁₁ , Xaa ₁₂ , Xaa ₁₃ , Xaa ₁₄ , Xaa ₁₅ , Xaa ₁₆ , Xaa ₁₇ , Xaa ₁₉ , Xaa ₂₀ , Xaa ₂₁ , Xaa ₂₄ , Xaa ₂₅ , Xaa ₂₆ , Xaa ₂₇ and Xaa ₂₈ are Ala; or provided that if Xaa ₁ is His, Arg or Tyr, then at least one of Xaa ₃ , Xaa ₄ and Xaa ₉ is Ala. Some are included. A particularly preferred compound of formula (II) is the application number PCT / US98 / 24273 filed November 13, 1998 entitled "Novel Exendin Agonist Compounds" having the amino acid sequence of SEQ ID NO: 5-93. Including those described in.

According to a particularly preferred embodiment, formula (II) wherein Xaa ₁₄ is Ala, Leu, Ile, Val or pentylglycine, more preferably Leu or pentylglycine, and Xaa ₂₅ is Ala, Phe, Tyr. Or naphthylalanine, more preferably Phe or naphthylalanine]. These compounds will be less sensitive to oxidative degradation reactions both in vitro and in vivo, and during the synthesis of the compounds.

Formula III
Also within the scope of the invention are the narrower genus concepts of compounds having peptides of various lengths, such as, for example, the genus concept of compounds that do not include peptides having a length of 28, 29 or 30 amino acid residues, respectively. is there. In addition, the present invention provides, for example, formula (III) [SEQ ID NO: 22]:
Xaa ₁ Xaa ₂ Xaa ₃ Gly Xaa ₅ Xaa ₆ Xaa ₇ Xaa ₈ Xaa ₉ Xaa ₁₀
Xaa ₁₁ Xaa ₁₂ Xaa ₁₃ Xaa ₁₄ Xaa ₁₅ Xaa ₁₆ Xaa ₁₇ Ala Xaa ₁₉
Xaa ₂₀ Xaa ₂₁ Xaa ₂₂ Xaa ₂₃ Xaa ₂₄ Xaa ₂₅ Xaa ₂₆ Xaa ₂₇ Xaa ₂₈ -Z ₁ ;
[Where:
Xaa ₁ 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;
Xaa ₁₀ is Ala, Leu or pentylglycine;
Xaa ₁₁ is Ala or Ser;
Xaa ₁₂ is Ala or Lys;
Xaa ₁₃ is Ala or Gln;
Xaa ₁₄ is Ala, Leu or pentylglycine;
Xaa ₁₅ is Ala or Glu;
Xaa ₁₆ is Ala or Glu;
Xaa ₁₇ is Ala or Glu;
Xaa ₁₉ is Ala or Val;
Xaa ₂₀ is Ala or Arg;
Xaa ₂₁ is Ala or Leu;
Xaa ₂₂ is Phe or naphthylalanine;
Xaa ₂₃ is Ile, 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;
Z ₁ is -OH,
-NH ₂ ,
Gly-Z ₂ ,
Gly Gly -Z ₂ ,
Gly Gly Xaa ₃₁ -Z ₂ ,
Gly Gly Xaa ₃₁ Ser-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ -Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ -Z ₂ or
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ Xaa ₃₈ -Z ₂ ;
Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ and Xaa ₃₈ are independently selected from Pro, homoproline, thioproline and N-methylylalanine; and
Z ₂ is -OH or -NH ₂ ;
However, Xaa ₃ , Xaa ₅ , Xaa ₆ , Xaa ₈ , Xaa ₁₀ , Xaa ₁₁ , Xaa ₁₂ , Xaa ₁₃ , Xaa ₁₄ , Xaa ₁₅ , Xaa ₁₆ , Xaa ₁₇ , Xaa ₁₉ , Xaa ₂₀ , Xaa ₂₁ , Xaa ₂₄ , Xaa ₂₅ , Xaa ₂₆ , Xaa ₂₇ and Xaa ₂₈ , within 3 are Ala]; and pharmaceutically acceptable salts thereof, entitled “Novel Exendin Agonist Compounds” having a specific amino acid sequence It includes a narrower genus concept for the compounds described in PCT Application No. PCT / US98 / 24210 filed November 13, 1998.

Formula IV
In addition, the present invention provides, for example, formula [IV] [SEQ ID NO: 23]:
Xaa ₁ Xaa ₂ Xaa ₃ Xaa ₅ Xaa ₅ Xaa ₆ Xaa ₇ Xaa ₈ Xaa ₉ Xaa ₁₀
Xaa ₁₁ Xaa ₁₂ Xaa ₁₃ Xaa ₁₄ Xaa ₁₅ Xaa ₁₆ Xaa ₁₇ Ala Xaa ₁₉
Xaa ₂₀ Xaa ₂₁ Xaa ₂₂ Xaa ₂₃ Xaa ₂₄ Xaa ₂₅ Xaa ₂₆ Xaa ₂₇ Xaa ₂₈ -Z ₁ ;
[Where:
Xaa ₁ 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;
Xaa ₁₀ is Ala, Leu or pentylglycine;
Xaa ₁₁ is Ala or Ser;
Xaa ₁₂ is Ala or Lys;
Xaa ₁₃ is Ala or Gln;
Xaa ₁₄ is Ala, Leu, Met or pentylglycine;
Xaa ₁₅ is Ala or Glu;
Xaa ₁₆ is Ala or Glu;
Xaa ₁₇ is Ala or Glu;
Xaa ₁₉ is Ala or Val;
Xaa ₂₀ is Ala or Arg;
Xaa ₂₁ is Ala or Leu;
Xaa ₂₂ is Phe or naphthylalanine;
Xaa ₂₃ is Ile, 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;
Z ₁ is -OH,
-NH ₂ ,
Gly-Z ₂ ,
Gly Gly-Z ₂
Gly Gly Xaa ₃₁ -Z ₂ ,
Gly Gly Xaa ₃₁ Ser-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ -Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ -Z ₂
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ Xaa ₃₈ -Z ₂
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ Xaa ₃₈ Ser-Z ₂ ;
Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ and Xaa ₃₈ are independently Pro, homoproline, thioproline, or N-methylylalanine; and
Z ₂ is -OH or -NH ₂ ;
However, Xaa ₃ , Xaa ₅ , Xaa ₆ , Xaa ₈ , Xaa ₁₀ , Xaa ₁₁ , Xaa ₁₂ , Xaa ₁₃ , Xaa ₁₄ , Xaa ₁₅ , Xaa ₁₆ , Xaa ₁₇ , Xaa ₁₉ , Xaa ₂₀ , Xaa ₂₁ , Xaa ₂₄ , And no more than three of Xaa ₂₅ , Xaa ₂₆ , Xaa ₂₇ , and Xaa ₂₈ are Ala; and if Xaa ₁ is His, Arg or Tyr, then at least one of Xaa ₃ , Xaa ₄ and Xaa ₉ One is Ala]; and a PCT application number filed on November 13, 1998 entitled "Novel Exendin Agonist Compounds" having a specific amino acid sequence such as: Includes a narrower genus concept of peptide compounds described in PCT / US98 / 24273.

Preferred compounds of formula (IV) include those where Xaa ₁ is His, Ala, Norval or 4-imidazopropionyl. Preferably Xaa ₁ is His, or 4-imidazopropionyl or Ala, more preferably His or 4-imidazopropionyl.

  Preferred compounds of formula (IV) include those where Xaa2 is Gly.

  Preferred compounds of formula (IV) include those where Xaa4 is Ala.

  Preferred compounds of formula (IV) include those where Xaa9 is Ala.

  Preferred compounds of formula (IV) include those where Xaa14 is Leu, pentylglycine or Met.

  Preferred compounds of formula (IV) include those where Xaa25 is Trp or Phe.

  Preferred compounds of formula (IV) include those where Xaa6 is Ala, Phe or naphthylalanine; Xaa22 is Phe or naphthylalanine; and Xaa23 is Ile or Val.

  Preferred compounds of formula (IV) include those wherein Z1 is -NH2.

  Preferred compounds of formula (IV) 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 (IV) include those wherein Xaa39 is Ser or Tyr, preferably Ser.

  Preferred compounds of formula (IV) include those where Z2 is -NH2.

  Preferred compounds of formula (IV) include those wherein Z1 is -NH2.

  Preferred compounds of formula (IV) include those where Xaa21 is Lys-NH □ -R, wherein R is Lys, Arg, C1-C10 linear or branched alkanoyl.

  Preferred compounds of formula (IV) are those in which X1 is Lys Asn, Lys-NHε-R Asn, or Lys-NHε-R Ala, where R is Lys, Arg, C1-C10 linear or branched alkanoyl. Including some. Preferred compounds of formula (IV) are those described in PCT Application No. PCT / US98 / 24273 filed November 13, 1998 entitled "Novel Exendin Agonist Compounds" selected from SEQ ID NOs: 95-110. Including those having an amino acid sequence.

Formula V
Also, the formula (V) [SEQ ID NO: 24]:
Xaa ₁ Xaa ₂ Xaa ₃ Gly Xaa ₅ Xaa ₆ Xaa ₇ Xaa ₈ Xaa ₉ Xaa ₁₀
Xaa ₁₁ Xaa ₁₂ Xaa ₁₃ Xaa ₁₄ Xaa ₁₅ Xaa ₁₆ Xaa ₁₇ Ala Xaa ₁₉ Xaa ₂₀
Xaa ₂₁ Xaa ₂₂ Xaa ₂₃ Xaa ₂₄ Xaa ₂₅ Xaa ₂₆ X ₁ -Z ₁ ;
[Where:
Xaa ₁ 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;
Xaa ₁₀ is Ala, Leu, Ile, Val, pentylglycine or Met;
Xaa ₁₁ is Ala or Ser;
Xaa ₁₂ is Ala or Lys;
Xaa ₁₃ is Ala or Gln;
Xaa ₁₄ is Ala, Leu, Ile, pentylglycine, Val or Met;
Xaa ₁₅ is Ala or Glu;
Xaa ₁₆ is Ala or Glu;
Xaa ₁₇ is Ala or Glu;
Xaa ₁₉ is Ala or Val;
Xaa ₂₀ is Ala or Arg;
Xaa ₂₁ is Ala, Leu or Lys-NH ^ε -R, wherein R is Lys, Arg, C ₁ -C ₁₀ linear or branched alkanoyl or cycloalkylalkanoyl;
Xaa ₂₂ is Phe, Tyr or naphthylalanine;
Xaa ₂₃ is Ile, 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;
X ₁ is Lys Asn, Asn Lys, Lys-NH ^ε -R Asn, Asn Lys-NH ^ε -R, Lys-NH ^ε -R Ala, Ala Lys-NH ^ε -R (where R is Lys, Arg C ₁ -C _{10 are} straight-chain or branched alkanoyl or cycloalkylalkanoyl),
Z ₁ is -OH,
-NH ₂ ,
Gly-Z ₂ ,
Gly Gly-Z ₂ ,
Gly Gly Xaa ₃₁ -Z ₂ ,
Gly Gly Xaa ₃₁ Ser-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ -Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ -Z ₂ or
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ Xaa ₃₈ -Z ₂ ;
Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ and Xaa ₃₈ are independently selected from the group consisting of Pro, homoproline, 3Hyp, 4Hyp, thioproline, N-alkyl glycine, N-alkylpentyl glycine and N-alkyl alanine ; And
Z ₂ is -OH or -NH ₂ ;
However, Xaa ₃ , Xaa ₅ , Xaa ₆ , Xaa ₈ , Xaa ₁₀ , Xaa ₁₁ , Xaa ₁₂ , Xaa ₁₃ , Xaa ₁₄ , Xaa ₁₅ , Xaa ₁₆ , Xaa ₁₇ , Xaa ₁₉ , Xaa ₂₀ , Xaa ₂₁ , Xaa ₂₄ , Description of PCT Application No. PCT / US98 / 24210 filed on November 13, 1998 entitled "Novel Exendin Agonist Compounds", including Xaa ₂₅ and Xaa ₂₆ within 3 of Ala] Are provided. Also within the scope of the invention are pharmaceutically acceptable salts and pharmaceutical compositions of the compound of formula (V) containing the compound and its salts.

Preferred exendin analogs of formula (V) include those where Xaa ₁ is His, Tyr or 4-imidazopropionyl. More preferably, Xaa ₁ is His.

Preference is given to compounds of the formula (V) in which Xaa ₁ is 4-imidazopropionyl.

Compounds of formula (V) [wherein Xaa ₂ is Gly] are preferred.

Preferred compounds of formula (V) are those wherein Xaa ₁₄ is Leu, pentylglycine or Met.

Preferred compounds of formula (V) are those wherein Xaa ₂₅ is Trp or Phe.

According to one embodiment, compounds of formula (V) wherein Xaa ₆ is Phe or naphthylalanine; and Xaa ₂₂ is Phe or naphthylalanine; and Xaa ₂₃ is Ile or Val are preferred. More preferably, Z ₁ is —NH ₂ . According to one embodiment, of formula (V), wherein Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ and Xaa ₃₈ are independently selected from the group consisting of Pro, homoproline, thioproline and N-alkylalanine Such compounds are particularly preferred. More preferably, Z ₂ is —NH ₂ .

Preferred compounds of formula (V) are those wherein X ₁ is Lys Asn, Lys-NH ^ε -R Asn, or Lys-NH ^ε -R Ala [wherein R is Lys, Arg, C ₁ -C ₁₀ linear or It is a branched alkanoyl]. Preferred compounds of formula (V) are described in PCT Application No. PCT / US98 / 24210, filed November 13, 1998, identified as Compound No. 62-69 and entitled “Novel Exendin Agonist Compounds” Of the compound.

Preferred such exendin analogs include those where Xaa ₁ is His, Ala or Norval. More preferably, Xaa ₁ is His or Ala. Most preferably, Xaa ₁ is His.

Compounds of formula (V) [wherein Xaa ₂ is Gly] are preferred.

Compounds of formula (V) [wherein Xaa ₃ is Ala] are preferred.

Preference is given to compounds of the formula (V) [wherein Xaa ₄ is Ala].

Compounds of formula (V) wherein Xaa ₉ is Ala are preferred.

Compounds of formula (V) wherein Xaa ₁₄ is Leu, pentylglycine or Met are preferred.

Preferred compounds of formula (V) are those wherein Xaa ₂₅ is Trp or Phe.

Preferred compounds of formula (V) are those wherein Xaa ₆ is Ala, Phe or naphthylalanine; Xaa ₂₂ is Phe or naphthylalanine; and Xaa ₂₃ is Ile or Val.

Preference is given to compounds of the formula (V) in which Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ and Xaa ₃₈ are independently selected from Pro, homoproline, thioproline and N-alkylalanine.

Preferably Z ₁ is —NH ₂ .

Preferably Z ₂ is —NH ₂ .

According to one embodiment, the formula (V) wherein Xaa ₁ is Ala, His or Tyr, more preferably Ala or His; Xaa ₂ is Ala or Gly; Xaa ₆ is Phe or naphthylalanine Xaa ₁₄ is Ala, Leu, pentylglycine or Met; Xaa ₂₂ is Phe or naphthylalanine; Xaa ₂₃ is Ile or Val; Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ and Xaa ₃₈ Are independently selected from Pro, homoproline, thioproline or N-alkylalanine; and Xaa ₃₉ is Ser or Tyr, more preferably Ser]. More preferably, Z ₁ is —NH ₂ .

According to a particularly preferred embodiment, a particularly preferred compound is of formula (V) wherein Xaa ₁ 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 Xr ₉ is Ala, Asp or Glu; Xaa ₁₀ is Ala, Leu or pentylglycine; Xaa ₁₁ is Ala or Ser; Xaa ₁₂ is Ala or Lys; Xaa ₁₃ is Ala or Gln; Xaa ₁₄ is Ala, Leu, Met or pentylglycine; Xaa ₁₅ is Ala or Glu; Xaa ₁₆ is Ala or Glu; Xaa ₁₇ is Ala or Xlu ₁₉ is Ala or Val; Xaa ₂₀ is Ala or Arg; Xaa ₂₁ is Ala or Leu; Xaa ₂₂ is Phe or naphthylalanine; Xa a ₂₃ is Ile, 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; Z ₁ is -OH, -NH ₂ , Gly-Z ₂ , Gly Gly-Z ₂ , Gly Gly Xaa ₃₁ -Z ₂ , Gly Gly Xaa ₃₁ Ser-Z ₂ , Gly Gly Xaa ₃₁ Ser Ser-Z ₂ , Gly Gly Xaa ₃₁ Ser Ser Gly-Z ₂ , Gly Gly Xaa ₃₁ Ser Ser Gly Ala-Z ₂ , Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ -Z ₂ , Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ -Z ₂ , Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ Xaa ₃₈ -Z ₂ or Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ Xaa ₃₈ Xaa ₃₉ -Z ₂ ; Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ and Xaa ₃₈ are independently Pro homoproline, thioproline or N-methylalanine; and Z ₂ is --OH or --NH ₂ ; however, Xaa ₃ , Xaa ₅ , Xaa ₆ , Xaa ₈ , Xaa ₁₀ , Xaa ₁₁ , Xaa ₁₂ , Xaa ₁₃ , Xaa ₁₄ , Xa Within 3 of a ₁₅ , Xaa ₁₆ , Xaa ₁₇ , Xaa ₁₉ , Xaa ₂₀ , Xaa ₂₁ , Xaa ₂₄ , Xaa ₂₅ , Xaa ₂₆ , Xaa ₂₇ and Xaa ₂₈ is Ala; and Xaa ₁ is If His, Arg or Tyr, then at least one of Xaa ₃ , Xaa ₄ and Xaa ₉ is Ala]. A particularly preferred compound of formula (V) is PCT Application No. PCT / US98 filed November 13, 1998 entitled "Novel Exendin Agonist Compounds" having the amino acid sequence identified as SEQ ID NO: 5-93. Includes those described in / 24210.

  According to a particularly preferred embodiment, the formula (V) wherein Xaa14 is Ala, Leu, Ile, Val or pentylglycine, more preferably Leu or pentylglycine, and Xaa25 is Ala, Phe, Tyr or naphthylalanine, more preferably Is Phe or naphthylalanine]. These compounds will be less sensitive to oxidative degradation reactions both in vitro and in vivo and during the synthesis of the compounds.

Formula VI
Also, the formula (VI) [SEQ ID NO: 25]:
Xaa ₁ Xaa ₂ Xaa ₃ Xaa ₄ Xaa ₅ Xaa ₆ Xaa ₇ Xaa ₈ Xaa ₉ Xaa ₁₀
Xaa ₁₁ Xaa ₁₂ Xaa ₁₃ Xaa ₁₄ Xaa ₁₅ Xaa ₁₆ Xaa ₁₇ Ala Xaa ₁₉ Xaa ₂₀
Xaa ₂₁ Xaa ₂₂ Xaa ₂₃ Xaa ₂₄ Xaa ₂₅ Xaa ₂₆ X ₁ -Z ₁ ;
[Where:
Xaa ₁ is His, Arg, Tyr, Ala, Norval, Val, Norleu or 4-imidazopropionyl;
Xaa ₂ is Ser, Gly, Ala or Thr;
Xaa ₃ is Ala, Asp or Glu;
Xaa ₄ 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;
Xaa ₁₀ is Ala, Leu, Ile, Val, pentylglycine or Met;
Xaa ₁₁ is Ala or Ser;
Xaa ₁₂ is Ala or Lys;
Xaa ₁₃ is Ala or Gln;
Xaa ₁₄ is Ala, Leu, Ile, pentylglycine, Val or Met;
Xaa ₁₅ is Ala or Glu;
Xaa ₁₆ is Ala or Glu;
Xaa ₁₇ is Ala or Glu;
Xaa ₁₉ is Ala or Val;
Xaa ₂₀ is Ala or Arg;
Xaa ₂₁ is Ala, Leu or Lys-NH ^ε -R where R is Lys, Arg, C ^1-10 linear or branched alkanoyl or cycloarray-alkanoyl;
Xaa ₂₂ is Phe, Tyr or naphthylalanine;
Xaa ₂₃ is Ile, 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;
X ₁ is Lys Asn, Asn Lys, Lys-NH ^ε -R Asn, Asn Lys-NH ^ε -R, Lys-NH ^ε -R Ala, Ala Lys-NH ^ε -R, wherein R is Lys , Arg, C ₁ -C _{10 are} straight or branched alkanoyl or cycloalkylalkanoyl)
Z ₁ is -OH,
-NH ₂ ,
Gly-Z ₂ ,
Gly Gly-Z ₂ ,
Gly Gly Xaa ₃₁ -Z ₂ ,
Gly Gly Xaa ₃₁ Ser-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala-Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ -Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ -Z ₂ ,
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ Xaa ₃₈ -Z ₂ or
Gly Gly Xaa ₃₁ Ser Ser Gly Ala Xaa ₃₆ Xaa ₃₇ Xaa ₃₈ Xaa ₃₉ -Z ₂ ;
Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ and Xaa ₃₈ 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 ;
However, Xaa ₃ , Xaa ₄ , Xaa ₅ , Xaa ₆ , Xaa ₈ , Xaa ₉ , Xaa ₁₀ , Xaa ₁₁ , Xaa ₁₂ , Xaa ₁₃ , Xaa ₁₄ , Xaa ₁₅ , Xaa ₁₆ , Xaa ₁₇ , Xaa ₁₉ , Xaa ₂₀ , Within 3 of Xaa ₂₁ , Xaa ₂₄ , Xaa ₂₅ , Xaa ₂₆ are Ala; and if Xaa ₁ is His, Arg, Tyr, or 4-imidazopropionyl, then Xaa ₃ , Xaa ₄ and Peptide described in PCT Application No. PCT / US98 / 24273 filed on November 13, 1998 entitled “Novel Exendin Agonist Compounds” comprising a compound of at least one of Xaa ₉ is Ala] A compound is also provided.

Preferred compounds of formula (VI) include those wherein Xaa ₁ is His, Ala, Norval or 4-imidazopropionyl. Preferably Xaa ₁ is His, or 4-imidazopropionyl or Ala, more preferably His or 4-imidazopropionyl.

Preferred compounds of formula (VI) include those where Xaa ₂ is Gly.

Preferred compounds of formula (VI) include those where Xaa ₄ is Ala.

Preferred compounds of formula (VI) include those where Xaa ₉ is Ala.

Preferred compounds of formula (VI) include those where Xaa ₁₄ is Leu, pentylglycine or Met.

Preferred compounds of formula (VI) include those where Xaa ₂₅ is Trp or Phe.

Preferred compounds of formula (VI) include those where Xaa ₆ is Ala, Phe or naphthylalanine; Xaa ₂₂ is Phe or naphthylalanine; and Xaa ₂₃ is Ile or Val.

Preferred compounds of formula (VI) include those where Z ₁ is —NH ₂ .

Preferred compounds of formula (VI) include those wherein Xaa ₃₁ , Xaa ₃₆ , Xaa ₃₇ and Xaa ₃₈ are independently selected from the group consisting of Pro, homoproline, thioproline and N-alkylalanine.

Preferred compounds of formula (VI) include those where Xaa ₃₉ is Ser or Tyr, preferably Ser.

Preferred compounds of formula (VI) include those where Z ₂ is —NH ₂ .

Preferred compounds of formula (VI) include those where Z ₁ is —NH ₂ .

Preferred compounds of formula (VI) include those where Xaa ₂₁ is Lys-NH ^ε -R, wherein R is Lys, Arg, C ₁ -C ₁₀ linear or branched alkanoyl.

Preferred compounds of formula (VI) are those wherein X ₁ is Lys Asn, Lys-NH ^ε -R Asn, or Lys-NH ^ε -R Ala, wherein R is Lys, Arg, C ₁ -C ₁₀ linear or branched That is alkanoyl].

  Preferred compounds of formula (VI) are PCT application numbers filed November 13, 1998 entitled "Novel Exendin Agonist Compounds" having an amino acid sequence selected from those identified as SEQ ID NOs: 95-110 Includes those described in PCT / US98 / 24273.

Formula VII
According to the invention, a particularly useful compound is of formula (VII) [SEQ ID NO: 26]:
Xaa ₁ Xaa ₂ Xaa ₃ Gly Thr Xaa ₄ Xaa ₅ Xaa ₆ Xaa ₇ Xaa ₈
Ser Lys Gln Xaa ₉ Glu Glu Glu Ala Val Arg Leu
Xaa ₁₀ Xaa ₁₁ Xaa ₁₂ Xaa ₁₃ Leu Lys Asn Gly Gly Xaa ₁₄
Ser Ser Gly Ala Xaa ₁₅ Xaa ₁₆ Xaa ₁₇ Xaa ₁₈ -Z
[Where:
Xaa ₁ is His, Arg or Tyr;
Xaa ₂ is Ser, Gly, Ala or Thr;
Xaa ₃ is Asp or Glu;
Xaa ₄ 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, Ile, Val, pentylglycine or Met;
Xaa ₉ is Leu, Ile, pentylglycine, Val or Met;
Xaa ₁₀ is Phe, Tyr or naphthalanine;
Xaa ₁₁ is Ile, Val, Leu, pentyl glycine, tert-butyl glycine or Met;
Xaa ₁₂ is Glu or Asp; Xaa ₁₃ is Trp, Phe, Tyr, or naphthylalanine;
Xaa ₁₄ , Xaa ₁₅ , Xaa ₁₆ and Xaa ₁₇ are independently Pro, homoproline, 3Hyp, 4Hyp, thioproline, N-alkyl glycine, N-alkylpentyl glycine or N-alkyl alanine;
Xaa ₁₈ is Ser, Thr or Tyr; and Z is —OH or —NH ₂ ;
However, the compound does not have any formula of SEQ ID NO: 7 or 9.] filed on January 7, 1998 entitled “Use of Exendins And Agonists There of For The Reduction of Food Intake” Exendin analogs having agonist activity described in US patent application Ser. No. 09 / 003,869. Preferred N-alkyl groups for N-alkylglycine, N-alkylpentylglycine and N-alkylalanine preferably include lower alkyl groups having 1 to about 6 carbon atoms, more preferably 1 to 4 carbon atoms. Also useful in the present invention are pharmaceutically acceptable salts of compounds of formula (VII).

Preferred exendin analogs include those where Xaa ₁ is His or Tyr. More preferably, Xaa ₁ is His.

A compound in which Xaa ₂ is Gly is preferred.

A compound in which Xaa ₉ is Leu, pentylglycine or Met is preferred.

Preferred compounds include those where Xaa ₁₃ is Trp or Phe.

Xaa ₄ is Phe or naphthalanine; Xaa ₁₁ is Il or Val, and Xaa ₁₄ , Xaa ₁₅ , Xaa ₁₆ and Xaa ₁₇ are independently selected from Pro, homoproline, thioproline or N-alkylalanine Is preferred. Preferably, the N-alkylalanine has an N-alkyl group of 1 to about 6 carbon atoms.

According to a particularly preferred embodiment, Xaa ₁₅ , Xaa ₁₆ and Xaa ₁₇ are the same amino acid residue.

Preferred are compounds wherein Xaa ₁₈ is Ser or Tyr, more preferably Ser.

Preferably Z is —NH ₂ .

According to one embodiment, formula (VII) wherein Xaa ₁ is His or Tyr, more preferably His; Xaa ₂ is Gly; Xaa ₄ is Phe or naphthalanine; Xaa ₉ is Xaa ₁₀ is Phe or naphthalanine; Xaa ₁₁ is Ile or Val; Xaa ₁₄ , Xaa ₁₅ , Xaa ₁₆ and Xaa ₁₇ are independently Pro, homoproline, and Leu, pentylglycine or Met; And thioproline or N-alkylalanine; and Xaa ₁₈ is Ser or Tyr, more preferably Ser]. More preferably Z is —NH ₂ .

According to a particularly preferred embodiment, a particularly preferred compound is of the formula (VII) wherein Xaa ₁ is His or Arg; Xaa ₂ is Gly; Xaa ₃ is Asp or Glu; Xaa ₄ is 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; Xaa ₁₀ is Phe or naphthylalanine; Xaa ₁₁ is Ile, Val or t-butyltylglycine; Xaa ₁₂ is Glu or Asp; Xaa ₁₃ Xaa ₁₄ , Xaa ₁₅ , Xaa ₁₆ , and Xaa ₁₇ are independently Pro, homoproline, thioproline, or N-methylalanine; Xaa ₁₈ is Ser or Tyr: and Z is —OH or —NH ₂ ; provided that the compound does not have any formula of SEQ ID NO: 7 or 9. I]. More preferably, Z is —NH ₂ .

According to a particularly preferred embodiment, there is provided a compound wherein Xaa ₉ is Leu, Ile, Val or pentylglycine, more preferably Leu or pentylglycine and Xaa ₁₃ is Phe, Tyr or naphthylalanine, more preferably Phe or naphthylalanine Is done. These compounds exhibit an advantageous duration of action and are believed to be less susceptible to oxidative degradation reactions both in vitro and in vivo, and during compound synthesis.

Formula VIII
Formula (VIII) [SEQ ID NO: 27]:
Xaa ₁ Xaa ₂ Xaa ₃ Gly Thr Xaa ₄ Xaa ₅ Xaa ₆ Xaa ₇ Xaa ₈
Ser Lys Gln Xaa ₉ Glu Glu Glu Ala Val Arg Leu
Xaa ₁₀ Xaa ₁₁ Xaa ₁₂ Xaa ₁₃ Leu X ₁ Gly Gly Xaa ₁₄
Ser Ser Gly Ala Xaa ₁₅ Xaa ₁₆ Xaa ₁₇ Xaa ₁₈ -Z
[Where:
Xaa ₁ 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 ₇ is Asp or Glu;
Xaa ₈ is Leu, Ile, Val, pentylglycine or Met;
Xaa ₉ is Leu, Ile, pentylglycine, Val or Met;
Xaa ₁₀ is Phe, Tyr or naphthylalanine;
Xaa ₁₁ is Ile, Val, Leu, pentyl glycine, tert-butyl glycine or Met;
Xaa ₁₂ is Glu or Asp;
Xaa ₁₃ is Trp, Phe, Tyr, or naphthylalanine; X ₁ is Lys Asn, Asn Lys, Lys-NH ^ε -R Asn, Asn Lys-NH ^ε -R (wherein R is Lys, Arg, which is a C ₁ -C ₁₀ linear or branched alkanoyl or cycloalkylalkanoyl);
Xaa ₁₄ , Xaa ₁₅ , Xaa ₁₆ and Xaa ₁₇ are independently Pro, homoproline, 3Hyp, 4Hyp, thioproline, N-alkyl glycine, N-alkylpentyl glycine or N-alkyl alanine;
Xaa ₁₈ is Ser, Thr or Tyr; Z is —OH or —NH ₂ ;
However, the compound does not have any formula of SEQ ID NO: 7 or 9.] PCT Application No. PCT / US98 / filed on August 6, 1998 entitled “Novel Exendin Agonist Compounds” The compounds described in 16387 are also provided. A suitable compound of formula (VIII) is PCT Application No. PCT / US98 / 16387 filed August 6, 1998 entitled "Novel Exendin Agonist Compounds" having the amino acid sequence of SEQ ID NOs: 37-40. Including the compounds described in 1.

Preferred exendin analogs of formula (VIII) include those where Xaa ₁ is His, Tyr or 4-imidazopropionyl. More preferably, Xaa ₁ is His or 4-imidazopropionyl.

Compounds of formula (VIII) [wherein Xaa ₂ is Gly] are preferred.

Preference is given to compounds of the formula (VIII) in which Xaa ₉ is Leu, pentylglycine or Met.

Preference is given to compounds of the formula (VIII) in which Xaa ₁₃ is Trp or Phe.

Formula (VIII) [wherein X ₁ is Lys Asn, or Lys-NH ^ε -R Asn (wherein R is Lys, Arg, C ₁ -C ₁₀ linear or branched alkanoyl)] Are preferred.

Further, the formula (VIII) [wherein, Xaa ₄ is Phe or naphthylalanine; Xaa ₁₀ is Phe or naphthylalanine; Xaa ₁₁ is Ile or _{Val, Xaa 14, Xaa 15,} Xaa 16 and Xaa ₁₇ Are independently selected from Pro, homoproline, thioproline or N-alkylalanine]. According to a particularly preferred embodiment, Xaa ₁₈ is Ser or Tyr. A compound in which Xaa ₁₈ is Ser is preferred. Preferably Z is —NH ₂ .

According to one preferred embodiment, formula (VIII): wherein Xaa ₄ is Phe or naphthylalanine; Xaa ₁₀ is Phe or naphthylalanine; Xaa ₁₁ is Ile or Val, X ₁ is Lys Asn, or Lys-NH ^ε -R Asn (wherein R is Lys, Arg, C ₁ -C ₁₀ linear or branched alkanoyl) and Xaa ₁₄ , Xaa ₁₅ , Xaa ₁₆ and Xaa ₁₇ are Are independently selected from Pro, homoproline, thioproline or N-alkylalanine].

  In another embodiment, exendins and exendin analogs of the invention do not include the peptide of SEQ ID NOs: 7-13. In this embodiment, preferred exendin analogs include an analog of formula (I-VIII) provided that the analog does not include the peptide of SEQ ID NOs: 7-13.

  Exendins and exendin agonists, such as the exendin analogs described herein, are described in, for example, Eng, et al., J. Biol. Chem. 265: 2025962, 1990; Peptides that can be prepared via peptide purification as described in Eng, et al., J. Biol. Chem. 267: 740205, 1992. Alternatively, exendins and exendin agonists are described in Raufman, et al., J. Biol. Chem incorporated herein, for example, using standard stationary phase peptide synthesis techniques and automated or semi-automated peptide synthesizers. 267: 2143237, 1992), which can be prepared by methods known to those skilled in the art. The compounds making up the active ingredients in the formulations and dosages of the invention can be prepared using standard stationary phase peptide synthesis techniques and preferably using automated or semi-automated peptide synthesizers. Typically, using such techniques, the α-N-carbamoyl protected amino acid and the amino acid attached to the growing peptide chain on the resin are dicyclohexylcarbodiimide and 1-hydroxy in the presence of a base such as diisopropylethylamine. Coupling at room temperature in an inert solvent such as dimethylformamide, N-methylpyrrolidinone or methylene chloride in the presence of a binder such as benzotriazole. The α-N-carbamoyl protecting group is removed from the resulting peptide-resin using a reagent such as trifluoroacetic acid or piperidine and coupled with the next desired N-protected amino acid to be added to the peptide chain. Repeat the ring reaction. Suitable N-protecting groups are well known in the art, with t-butyloxycarbonyl (tBoc) and fluorenylmethoxycarbonyl (Fmoc) being preferred herein.

  Solvents, amino acid derivatives and 4-methylbenzhydryl-amine resins used in peptide synthesizers can be purchased from Applied Biosystems Inc. (Foster City, CA). The following side chain protected amino acids can be purchased from Applied Biosystems, Inc .: BSD-112344.1-Arg (Pmc), Boc-Thr (Bzl), Fmoc-Thr (t-Bu), Boc-Ser (Bzl), Fmoc -Ser (t-Bu), Boc-Tyr (BrZ), Fmoc-Tyr (t-Bu), Boc-Lys (Cl-Z), Fmoc-Lys (Boc), Boc-Glu (Bzl), Fmoc-Glu (t-Bu), Fmoc-His (Trt), Fmoc-Asn (Trt), and Fmoc-Gln (Trt). Boc-His (BOM) can be purchased from Applied Biosystems, Inc. or Bachem Inc. (Torrance, CA). Anisole, dimethyl sulfide, phenol, ethanedithiol, and thioanisole are available from Aldrich Chemical Company (Milwaukee, Wis.). Air Products and Chemicals (Allentown, PA) supplies HF. Ethyl ether, acetic acid and methanol can be purchased from Fisher Scientific (Pittsburgh, PA).

  Stationary phase peptide synthesis is performed using the NMP / HOBt (Option 1) system and tBoc or Fmoc chemistry (Applied Biosystems User's Manual for the ABI 430A Peptide Synthesizer, Version 1.3B July 1, 1988, section 6, pp. 49-70, Applied Biosystems , Inc., Foster City, CA) and can be performed on an automated peptide synthesizer. Boc-peptide-resins can be cleaved with HF (-50 ° C. to 0 ° C., 1 hour). The peptide can be extracted from the resin alternately with water and acetic acid and the filtrate can be lyophilized. Fmoc-peptide resins can be cleaved by standard methods (Introduction to Cleavage Techniques, Applied Biosystems, Inc., 1990, pp. 6-12). Peptides can also be assembled using Advanced Chem Tech Synthesizer (Model MPS 350, Louisville, Kentucky).

Peptides can be purified by RP-HPLC (preparative and analytical) using a Waters Delta Prep 3000 system. C4, C8 or C18 preparative columns (10μ, 2.2 x 25 cm; Vydac, Hesperia, CA) can be used to isolate the peptides, and the purity is determined by C4, C8 or C18 analytical columns (5μ, 0.46 x 25 cm; Vydac). Solvents (A = 0.1% TFA / water and B = 0.1% TFA / CH ₃ CN) can be delivered to an analytical column with a flow rate of 1.0 ml / min and a preparation column of 15 ml / min. Amino acid analysis can be performed on the Waters Pico Tag system and processed using the Maxima program. Peptides can be hydrolyzed by gas phase acid hydrolysis (115 ° C., 20-24 hours). The hydrolyzate can be derivatized and analyzed by standard methods (Cohen, et al., The Pico Tag Method: A Manual of Advanced Techniques for Amino Acid Analysis, pp. 11-52, Millipore Corporation, Milford, MA (1989)). Fast atom bombardment analysis can be performed by M-Scan, Incorporated (West Chester, PA). Mass calibration can be performed using cesium iodide or cesium iodide / glycerol. Plasma desorption ionization analysis using time of mass detection can be performed on an Applied Biosystems Bio-Ion 20 mass spectrometer. Electrospray mass spectroscopy can be performed on a VG-Trio machine.

  Exendins and exendin agonists are peptides that can also be prepared using recombinant DNA technology, methods currently known in the art. See, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, 2d Ed., Cold Spring Harbor (1989). Alternatively, such compounds can be prepared by homogeneous phase peptide synthesis methods. Non-peptide compounds useful in the present invention can be prepared by methods known in the art. For example, phosphate-containing amino acids and peptides containing such amino acids can be prepared by methods known in the art. See, for example, Bartlett and Landen, Biorg. Chem. 14: 356-377 (1986).

Methods of the Invention The methods and compositions of the invention can be used to treat PCOS. Many of the symptoms associated with PCOS are due to underlying insulin resistance. PCOS related symptoms include insulin resistance, hyperinsulinemia, hyperandrogenism, type 2 diabetes, irregular menstruation, anovulation and infertility. Accordingly, the present invention provides a method of treating insulin resistance in a subject suffering from PCOS comprising the step of administering GLP-1.

  Insulin resistance can be an abnormal receptor precursor (e.g., abnormal ligand or competition), receptor (e.g., abnormal structure, affinity of ligand for receptor, or number of receptors), or post-receptor (e.g., It can be the result of any one or more events including (abnormal signaling) events. Insulin resistance can be determined by a number of methods known in the art. For example, insulin resistance can be diagnosed using normoglycemic hyperinsulinic clamp technology (Rao, G., Am. Fam. Physician (2001) 63: 1159-63). This technique involves administering an insulin dose intravenously by also administering glucose while simultaneously maintaining the glucose at a preset level within the normal range. At equilibrium, the amount of glucose uptake by a particular tissue in the presence of a particular dose of insulin can be calculated. Other methods used to detect insulin resistance include insulin suppression test, intravenous glucose tolerance test, and continuous infusion of glucose by model evaluation (Rao, G., above).

  In another embodiment, the present invention provides a method for preventing the onset of type 2 diabetes in a subject suffering from PCOS comprising administering GLP-1, exendin, or an agonist or analog of these compounds To do. Type 2 diabetes is often the result of hyperinsulinemia caused by insulin resistance. Thus, treating insulin resistance in these patients will prevent the development of type 2 diabetes. Methods for diagnosing type 2 diabetes are well known in the art.

  In yet another embodiment, the present invention restores regular menstruation in a subject suffering from PCOS, comprising administering GLP-1, exendin, or agonists or analogs of these compounds. Methods are provided to restore ovulation and / or restore fertility. PCOS patients often present with androgen hypertrophy that is thought to be caused by hyperinsulinemia. Androgen hypertrophy results in follicular retraction, anovulation, and infertility. Thus, by administering GLP-1, exendin or agonists or analogs of these compounds, reducing insulin resistance ameliorates hyperinsulinemia, thereby providing regular menstruation, ovulation, and / or Or fertility can be restored.

  In another embodiment, the present invention relates to GLP-1, exendin or agonists or analogs of these compounds, and drugs that induce ovulation (clomiphene, follistim, and gonal-F (GonalF)). Together, a method of treating PCOS is provided, characterized by co-administration to a patient in need thereof.

  In yet another embodiment, the present invention relates to GLP-1, exendin or agonists or analogs of these compounds, oral contraceptives (eg, progestogens and estrogens), spironolactone, flutamide, and finasteride. A method of treating PCOS, characterized in that it is co-administered to a patient in need thereof with an antiandrogen, including but not limited to (finasteride).

  In yet another embodiment, the present invention relates to GLP-1, exendin or agonists or analogs of these compounds, metformin, D-chiro-inositol diazoxide, and PPAR inhibitors. (Eg, but not limited to insulin, including, but not limited to, troglitazone (Rizulin), rosiglitazone (Avandia), and pioglitazone (Actos)) Provided is a method of treating PCOS characterized by co-administration to a patient in need thereof with a sensitizer.

  In another embodiment, the present invention treats PCOS characterized by co-administering GLP-1, exendin or an agonist or analog of these compounds together with glucose to a patient in need thereof Provide a way to do it. In a more preferred embodiment, glucose is administered intravenously.

  When the compounds of the invention are administered in combination therapy, as described above, they may be administered to the patient sequentially or simultaneously. Alternatively, the pharmaceutical composition of the present invention may comprise a combination of these compound molecules and another agent GLP-1, exendin, or agonist or analog as described above.

  In a preferred embodiment, the subject afflicted with PCOS is a mammal such as a dog, cat, rodent. In a more preferred embodiment, the subject afflicted with PCOS is a human.

Pharmaceutical composition
GLP-1, exendin, and agonists or analogs of these compounds can be formulated into pharmaceutical compositions for administration to subjects, including humans. These pharmaceutical compositions are preferably GLP-1 effective in treating a subject suffering from PCOS, for example to treat insulin resistance, prevent type 2 diabetes, and treat regular menstruation and / or infertility. , Exendins or agonists or analogs of these compounds, and a pharmaceutically acceptable carrier.

  Pharmaceutically acceptable carriers useful in these pharmaceutical compositions include, for example, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphate, glycine, Sorbic acid, potassium sorbate, partial glyceride mixture of saturated vegetable fatty acids, electrolytes such as water, salt or protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, colloidal silica, three Magnesium silicate, polyvinyl pyrrolidone, cellulosic material, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylate, wax, polyethylene-polyoxypropylene-block polymer, polyethylene glycol and wool fat.

  The compositions of the invention can be administered parenterally, orally, by suction spray, topically, rectally, nasally, buccally, vaginally or via an implanted container. Can be administered. As used herein, the term “parenteral” refers to subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intradermal, intrahepatic, intralesional and intracranial injection or infusion techniques. including. Preferably, the composition is administered by infusion pump or subcutaneous injection of a sustained release formulation.

  Sterile injectable forms of the compositions of this invention may be aqueous or oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be used are water, Ringer's solution and physiological saline. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. Fatty acids such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, especially their polyoxyethylated versions, like natural pharmaceutically acceptable oils such as olive oil or castor oil . These oily solutions or suspensions can also be diluted with long chain alcohols such as carboxymethylcellulose or similar dispersants commonly used in the formulation of pharmaceutically acceptable dosage forms including emulsions and suspensions. Agents or dispersants can be included. Other commonly used surfactants such as Tweens, Spans and bioavailability enhancers commonly used in the manufacture of pharmaceutically acceptable solids, liquids, other emulsifiers or other dosage forms Agents can also be used for formulation purposes.

  A parenteral formulation can be a single bolus dose, an infusion or loading bolus dose, followed by a maintenance dose. These compositions may be administered once a day or “as needed”.

  The pharmaceutical compositions of the invention can be administered orally in any orally acceptable dosage form including capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, carriers that are commonly used include lactose and corn starch. Lubricants such as magnesium stearate are also commonly added. For oral administration in a capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring, or coloring agents may be added.

  Alternatively, the pharmaceutical compositions of the present invention can be administered in the form of suppositories for rectal administration. These can be prepared by mixing the agent with a suitable nonirritating excipient that is solid at room temperature but liquid at rectal temperature and therefore melts in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.

  The pharmaceutical composition of the present invention can also be administered locally. Topical application can be carried out in a rectal suppository formulation (see above) or in a suitable enema formulation.

  For topical application, the pharmaceutical compositions may be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Carriers for topical application of the compounds of the invention include mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutical composition can be formulated in a suitable lotion or cream containing the active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-oxyldodecanol, benzyl alcohol, and water.

  For ophthalmic use, the pharmaceutical composition is as a finely divided suspension in isotonic, pH-adjusted sterile saline, with or without preservatives such as benzalkonium chloride, or Preferably, it can be formulated as an isosmotic, pH-adjusted sterile physiological saline. Alternatively, for ophthalmic use, the pharmaceutical composition may be formulated in an ointment such as petrolatum.

  The pharmaceutical composition of the present invention can also be administered by nasal aerosol or inhalation. Such compositions are prepared by techniques well known in the art of pharmaceutical formulation and are used to enhance bioavailability, fluorocarbons, and / or other conventional solubilizers or dispersants, such as benzyl alcohol or other Can be prepared as a solution in saline using a suitable preservative, absorption promoter.

  The amount of GLP-1, exendin, or an agonist or analog of these compounds that can be combined with a carrier material to produce a single dosage form will vary depending on the host being treated and the particular mode of administration. . The composition is administered to a patient receiving a dose of GLP-1, exendin, or an agonist or analog of these compounds at a dose between 0.1-1000 pmoles / kg body weight per minute (when administered by infusion). Can be prescribed as In some embodiments of the invention, the dosage is 1-10 pmoles / kg body weight / minute (when administered by infusion). In a preferred embodiment, when administered by intravenous infusion, the dose is 0.5-2.0 pmoles / kg / min. The composition may be administered over a single dose, multiple doses, or an established period of infusion.

  In preferred embodiments, GLP-1, exendin, or agonists or analogs of these compounds are administered to patients with chronic polycystic ovary syndrome. In another preferred embodiment, GLP-1, exendin, or an agonist or analog of these compounds is administered at least once a day or by continuous infusion via a pump. In yet another preferred embodiment, GLP-1, exendin, or an agonist or analog of these compounds is formulated by administration from subcutaneous deposits, oral administration or intermittent aspiration over a period of days to weeks. Is done.

  Specific doses and treatment plans for any particular patient are specific GLP-1, exendin, or agonists or analogs of these compounds, patient age, weight, general health, sex, and diet, and It will depend on a variety of factors including the time of administration, the rate of excretion, the drug combination, and the severity of the particular disease being treated. The determination of such factors by medical caregivers is within the ordinary skill in the art. The amount of GLP-1, exendin, or agonists or analogs of these compounds can also be determined by the individual patient being treated, the route of administration, the type of formulation, the characteristics of the compound used, the severity of the disease, and the desired Will be determined by the effect of. The amount of GLP-1, exendin, or agonist or analog of these compounds can be determined by pharmacological and pharmacokinetic principles well known in the art.

  In order that the invention described herein may be more fully understood, the following examples are set forth. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting the invention in any way.

Example
1. Treatment of PCOS patients with GLP-1
PCOS patients are women only. Typically, pre-menopausal women exhibit diseases with irregular menstruation, infertility, excessive hair growth, acne and hair loss. Postmenopausal women can have all of these illnesses except irregular menstruation. Obesity, hypertension and diabetes are disorders commonly associated with PCOS.

  The diagnosis of PCOS will be confirmed by measuring the level of serum testosterone and / or the ratio of serum LH / FSH. An elevated level of serum testosterone (> 60 ng / ml) or an abnormal serum LH / FSH ratio (<2.5) is a sign of PCOS.

  Chronic PCOS patients will be treated with GLP-1. GLP-1 will be administered by one or more injections per day or by continuous infusion through a pump that delivers a certain amount of drug. Alternatively, GLP-1 may be formulated for intermittent aspiration or oral administration from subcutaneous deposits over days to weeks.

  Regardless of the mode of administration, the total amount of GLP-1 delivered to the blood of a PCOS patient will be in the range of 720 to 2880 pmol / kg / day. This is equivalent to 0.5-2.0 pmoles / kg / min when administered by intravenous infusion.

  The effects of GLP-1 may improve or improve major illnesses, including but not limited to normalizing menstruation, restoring fertility, excessive hair loss, acne elimination and hair loss cessation. Will be established by deciding. Other indicators of GLP-1 effect may be used, including but not limited to serum testosterone levels and LH / FSH ratio. The GLP-1 effect will be determined by a decrease in serum testosterone levels and an increase in the LH / FSH ratio.

[Sequence Listing]

Claims (64)

  Administering a compound selected from the group consisting of exendin having the sequences set forth in SEQ ID NOs: 14-27, and agonists and analogs thereof, wherein the subject exhibits at least one symptom of PCOS, How to treat a subject suffering from.   The symptoms include insulin resistance, hyperinsulinemia, type 2 diabetes, obesity, hypertension, hyperlipidemia, anovulation or irregular ovulation, infertility, hyperandrogenism, hirsutism, alopecia, acne, hypertrophy The method of claim 1, wherein the method is selected from the group consisting of follicular ovary, abnormal uterine bleeding, and spontaneous abortion.   2. The method of claim 1, wherein the subject is a human.   2. The method of claim 1, wherein the exendin, or agonist or analog thereof, is administered by an infusion pump or by subcutaneous injection of a sustained release formulation.   The method according to claim 1, wherein the exendin, or an agonist or analog thereof, is administered with an agent selected from the group consisting of an ovulation inducer, an antiandrogen, an insulin sensitizer and glucose.   6. The method of claim 5, wherein the ovulation inducing agent is selected from the group consisting of clomiphene, follistim, and gonal-F.   6. The method of claim 5, wherein the antiandrogen is selected from the group consisting of oral contraceptives, spironolactone, flutamide, and finasteride.   Insulin sensitizers include metformin, D-chiro-inositol, diazoxide, and PPAR inhibitors (eg, troglitazone (Rizulin), rosiglitazone (Avantia) 6. The method of claim 5 selected from the group consisting of (Avandia)), and pioglitazone (Actos).   6. The method of claim 5, wherein glucose is administered intravenously.   6. The method of claim 5, wherein exendin, or an agonist or analog thereof, is administered either sequentially or simultaneously.   A method for reducing insulin resistance in a subject suffering from PCOS comprising the step of administering an exendin having a sequence set forth in SEQ ID NOs: 14-27 and a compound selected from the group consisting of agonists and analogs thereof.   12. The method of claim 11, wherein the subject is a human.   12. The method of claim 11, wherein exendin, or an agonist or analog thereof, is administered by an infusion pump or by subcutaneous injection of a sustained release formulation.   12. The method of claim 11, wherein the exendin, or agonist or analog thereof, is administered with an agent selected from the group consisting of an ovulation inducing agent, an antiandrogen agent, an insulin sensitizer and glucose.   15. The method of claim 14, wherein the ovulation inducing agent is selected from the group consisting of clomiphene, follistim, and gonal-F.   15. The method of claim 14, wherein the antiandrogen is selected from the group consisting of oral contraceptives, spironolactone, flutamide, and finasteride.   Insulin sensitizers include metformin, D-chiro-inositol, diazoxide, and PPAR inhibitors (eg, troglitazone (Rizulin), rosiglitazone (Avantia) 15. The method of claim 14 selected from the group consisting of (Avandia)), and pioglitazone (Actos).   15. The method of claim 14, wherein glucose is administered intravenously.   15. The method of claim 14, wherein exendin, or an agonist or analog thereof, is administered either sequentially or simultaneously.   A method for preventing the onset of type 2 diabetes in a subject suffering from PCOS, comprising a step of administering an exendin having a sequence set forth in SEQ ID NOs: 14-27 and a compound selected from the group consisting of agonists and analogs thereof.   21. The method of claim 20, wherein the subject is a human.   21. The method of claim 20, wherein the exendin, or agonist or analog thereof, is administered by an infusion pump or by subcutaneous injection of a sustained release formulation.   21. The method of claim 20, wherein the exendin, or agonist or analog thereof, is administered with an agent selected from the group consisting of an ovulation inducing agent, an antiandrogen agent, an insulin sensitizer and glucose.   24. The method of claim 23, wherein the ovulation inducing agent is selected from the group consisting of clomiphene, follistim, and Gonal-F.   24. The method of claim 23, wherein the antiandrogen is selected from the group consisting of oral contraceptives, spironolactone, flutamide, and finasteride.   Insulin sensitizers include metformin, D-chiro-inositol, diazoxide, and PPAR inhibitors (eg, troglitazone (Rizulin), rosiglitazone (Avantia) 24. The method of claim 23 selected from the group consisting of (Avandia)), and pioglitazone (Actos).   24. The method of claim 23, wherein glucose is administered intravenously.   24. The method of claim 23, wherein the exendin, or agonist or analog thereof, is administered either sequentially or simultaneously.   A method for restoring regular menstruation in a subject suffering from PCOS, comprising administering an exendin having a sequence set forth in SEQ ID NOs: 14-27 and a compound selected from the group consisting of agonists and analogs thereof.   30. The method of claim 29, wherein the subject is a human.   30. The method of claim 29, wherein the exendin, or agonist or analog thereof, is administered by an infusion pump or by subcutaneous injection of a sustained release formulation.   30. The method of claim 29, wherein the exendin, or agonist or analog thereof, is administered with an agent selected from the group consisting of an ovulation inducing agent, an antiandrogen agent, an insulin sensitizer and glucose.   35. The method of claim 32, wherein the ovulation inducing agent is selected from the group consisting of clomiphene, follistim, and Gonal-F.   35. The method of claim 32, wherein the antiandrogen is selected from the group consisting of oral contraceptives, spironolactone, flutamide, and finasteride.   Insulin sensitizers include metformin, D-chiro-inositol, diazoxide, and PPAR inhibitors (eg, troglitazone (Rizulin), rosiglitazone (Avantia) 33. The method of claim 32, wherein the method is selected from the group consisting of (Avandia)), and pioglitazone (Actos).   40. The method of claim 32, wherein glucose is administered intravenously.   35. The method of claim 32, wherein exendin, or an agonist or analog thereof, is administered either sequentially or simultaneously.   A method for recovering regular ovulation in a subject suffering from PCOS, comprising administering an exendin having a sequence set forth in SEQ ID NOs: 14-27 and a compound selected from the group consisting of agonists and analogs thereof.   40. The method of claim 38, wherein the subject is a human.   40. The method of claim 38, wherein the exendin, or agonist or analog thereof, is administered by an infusion pump or by subcutaneous injection of a sustained release formulation.   39. The method of claim 38, wherein the exendin, or agonist or analog thereof, is administered with an agent selected from the group consisting of an ovulation inducing agent, an antiandrogen agent, an insulin sensitizer and glucose.   42. The method of claim 41, wherein the ovulation inducing agent is selected from the group consisting of clomiphene, follistim, and Gonal-F.   42. The method of claim 41, wherein the antiandrogen is selected from the group consisting of oral contraceptives, spironolactone, flutamide, and finasteride.   Insulin sensitizers include metformin, D-chiro-inositol, diazoxide, and PPAR inhibitors (eg, troglitazone (Rizulin), rosiglitazone (Avantia) 42. The method of claim 41, selected from the group consisting of (Avandia)), and pioglitazone (Actos).   42. The method of claim 41, wherein the glucose is administered intravenously.   42. The method of claim 41, wherein exendin, or an agonist or analog thereof, is administered either sequentially or simultaneously.   A method for restoring fertility in a subject suffering from PCOS, comprising administering an exendin having a sequence set forth in SEQ ID NOs: 14-27 and a compound selected from the group consisting of agonists and analogs thereof.   48. The method of claim 47, wherein the subject is a human.   48. The method of claim 47, wherein the exendin, or agonist or analog thereof, is administered by an infusion pump or by subcutaneous injection of a sustained release formulation.   48. The method of claim 47, wherein the exendin, or agonist or analog thereof, is administered with an agent selected from the group consisting of an ovulation inducing agent, an antiandrogen agent, an insulin sensitizer and glucose.   51. The method of claim 50, wherein the ovulation inducing agent is selected from the group consisting of clomiphene, follistim, and Gonal-F.   51. The method of claim 50, wherein the antiandrogen is selected from the group consisting of oral contraceptives, spironolactone, flutamide, and finasteride.   Insulin sensitizers include metformin, D-chiro-inositol, diazoxide, and PPAR inhibitors (eg, troglitazone (Rizulin), rosiglitazone (Avantia) 51. The method of claim 50 selected from the group consisting of (Avandia)), and pioglitazone (Actos).   51. The method of claim 50, wherein the glucose is administered intravenously.   51. The method of claim 50, wherein exendin, or an agonist or analog thereof, is administered either sequentially or simultaneously.   A method for preventing spontaneous abortion in a subject suffering from PCOS, comprising a step of administering an exendin having a sequence set forth in SEQ ID NOs: 14-27 and a compound selected from the group consisting of agonists and analogs thereof.   57. The method of claim 56, wherein the subject is a human.   57. The method of claim 56, wherein the exendin, or agonist or analog thereof, is administered by an infusion pump or by subcutaneous injection of a sustained release formulation.   57. The method of claim 56, wherein the exendin, or agonist or analog thereof, is administered with an agent selected from the group consisting of an ovulation inducing agent, an antiandrogen agent, an insulin sensitizer and glucose.   60. The method of claim 59, wherein the ovulation inducing agent is selected from the group consisting of clomiphene, follistim, and Gonal-F.   60. The method of claim 59, wherein the antiandrogen is selected from the group consisting of oral contraceptives, spironolactone, flutamide, and finasteride.   Insulin sensitizers include metformin, D-chiro-inositol, diazoxide, and PPAR inhibitors (eg, troglitazone (Rizulin), rosiglitazone (Avantia) 60. The method of claim 59 selected from the group consisting of (Avandia)), and pioglitazone (Actos).   60. The method of claim 59, wherein the glucose is administered intravenously. 60. The method of claim 59, wherein the exendin, or agonist or analog thereof, is administered either sequentially or simultaneously.