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`DESCRIPTION
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`MODIFIED EXENDINS AND EXENDIN AGONISTS
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`RELATED APPLICATIONS
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`This application claims priority to, and the benefit of,
`United States provisional patent application serial no.
`60/132,018, filed April 30, 1999, which application is hereby
`incorporated by reference in its entirity.
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`FIELD OF THE INVENTION
`The present invention relates to novel modified exendins and
`exendin agonists having an exendin or exendin agonist peptide
`linked to one or more polyethylene glycol polymers (or other
`15 molecular weight increasing agents), and related products and
`methods that are useful, for example, in the treatment of
`diabetes, including Type 1 and 2 diabetes, in the treatment of
`disorders which would be benefited by agents which modulate
`plasma glucose levels·,· and in the treatment of disorders which
`20 would be benefited by the administration of agents useful in
`modulating glucagon or triglyceride levels, or the rate of
`gastric emptying or food intake, including obesity, eating.
`disorders, and insulin-resistance syndrome.
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`BACKGROUND
`The following description includes information that may be
`useful in understanding the present invention.
`It is not an
`admission that any of the information provided herein is prior
`art to the presently claimed invention, nor that any of the
`30 publications specifically or implicitly referenced are prior art
`to that invention.
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`The exendins are peptides that are found in the salivary
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`secretions of the Gila monster and the Mexican Bearded Lizard,
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`reptiles that are endogenous to Arizona and Northern Mexico.
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`Exendin-3
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`[SEQ. ID. NO. 1] is present in the salivary secretions
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`5 of Heloderma horridum (Mexican Beaded Lizard), and exendin-4
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`[SEQ. ID. NO. 2] is present in the salivary secretions of
`Heloder.m suspectum (Gila monster) (Eng, J., et al., J. Biol.
`Chem., 265:20259-62, 1990;. Eng, J., et al., J. Biol. Chern.,
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`267:7402-05, 1992). The amino acid sequence of exendin-3 is
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`shown in Figure 1 .. The amino acid sequence of exendin-4 is shown
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`in Figure 2. Exendin-4 was first thought to be a (potentially
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`toxic) component of the venom.
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`It now appears that exendin-4 is
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`devoid of toxicity, and that it instead is made in salivary
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`glands in the Gila monster.
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`The exendins have some sequence similarity to several
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`members of the glucagon-like peptide family, with the highest
`(Goke, et
`homology, 53%, being to GLP-1[7-36]NH2 [SEQ. ID. NO. 3]
`al., J. Biol. Chem., 268:19650-55, 1993). GLP-1[7-36]NH2, also
`sometimes referred to as proglucagon[78-107] or simply "GLP-1",
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`has an insulinotropic effect, stimulating insulin secretion from
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`pancreatic beta-cells; GLP-1 has also been reported to inhibit
`glucagon secretion from pancreatic alpha-cells (0rsov, et al.,
`Diabetes, 42:658-61, 1993; D'Alessio, et al., J. Clin. Invest.,
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`97:133-38, 1996). GLP-1 has been reported to inhibit gastric
`emptying (Willms B, et al., J. Clin. Endocrinol. Metab. 81 (1):
`327-32, 1996; Wettergren A, et al., Dig. Dis. Sci. 38 (4): 665-
`73, 1993), and gastric acid secretion (Schjoldager BT, et al.,
`Dig. Dis. Sci. 34 (5): 703-8, 1989; O'Halloran DJ, et al., J.
`Endocrinol. 126 (1): 169-73, 1990; Wettergren A, et al., Dig.
`30 Dis. Sci. 38 (4): 665-73, 1993)). GLP-1[7-37], which has an
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`additional glycine residue at -its carboxy terminus, is reported
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`to stimulate insulin secretion in humans (0rsov; et al.,
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`Diabetes, 42:658-61, 1993). Other reports relate to the
`inhibition of glucagon secretion (Creutzfeldt woe, et al.,
`Glucagonostatic actions and reduction of fasting hyperglycemia by
`exogenous glucagon-like peptide !(7-36) amide in Type 1 diabetic
`5 patients, Diabetes Care 1996;19(6) :580-6), and a purported role
`in appetite control (Turton MD, et al., A role for glucagon-like
`peptide-1 in the central regulation of feeding, Nature 1996
`Jan;379(6560) :69-72) .A transmembrane G-protein adenylate-cyclase(cid:173)
`coupled receptor, said to be responsible at least in part for the
`insulinotropic effect of GLP-1, has reportedly been cloned from a
`beta-cell line (Thorens, Proc. Natl. Acad. Sci. USA 89:8641-45,
`1992) . GLP-1 has been the focus of significant investigation in
`recent years due to its reported action on the amplification of
`stimulated insulin production (Byrne MM, Goke B. Lessons from
`human studies with glucagon-like peptide-1: Potential of the gut
`hormone for clinical use. In: Fehmann HC, Goke B. Insulinotropic
`Gut Hormone Glucagon-Like Peptide 1. Basel, Switzerland: Karger,
`1997 :219-33).
`GLP-1 has also been reported to restore islet glucose
`sensitivity in aging rats, restoring their glucose tolerance to
`that of younger rats (Egan JM, et al., Diabetologia 1997
`Jun;40(Suppl 1) :A130). However, the short duration of biological
`action of GLP-1 in vivo is one feature of the peptide that has
`hampered its development as a therapeutic agent. Various methods
`have been tried to prolong the half-life of GLP-1 or GLP-1(7-37),
`including attempts to alter their amino acid sequences and to
`deliver them using certain formulations (see, e.g., European
`Patent Application, entitled "Prolonged Delivery of Peptides," by
`Darley, et al., publication number 0 619 322 A2, regarding the
`inclusion of polyethylene glycol in formulations containing GLP-1
`(7-37)).
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`Pharmacological studies have led to reports that exendin-4
`can act at GLP-1 receptors in vitro on certain insulin-secreting
`cells, at dispersed acinar cells from guinea pig pancreas, and at
`parietal cells from stomach; the peptide is also reported to
`stimulate somatostatin release and inhibit gastrin release in
`isolated stomachs (Goke, et al., J. Biol. Chern. 268:19650-55,
`1993; Schepp, et al., Eur. J. Phar.macol., 69:183-91, 1994;
`Eissele, et al., Life Sci., 55:629-34, 1994). Exendin-3 and
`exendin-4 were reportedly found to stimulate cAMP production in,
`and amylase release from, pancreatic acinar cells (Malhotra, R.,
`et al., Regulatory Peptides, 41:149-56, 1992; Raufman, et al., J.
`Biol. Chern. 267:21432-37, 1992; Singh, et al., Regul. Pept.
`53:47-59, 1994). Exendin-4 has a significantly longer duration
`of action than GLP-1. For example, in one experiment, glucose
`lowering by exendin-4 in diabetic mice was reported to persist
`for several hours, and, depending on dose, for up to 24 hours
`(Eng, J. Prolonged effect of exendin-4 on hyperglycemia of db/db
`mice, Diabetes 1996 May; 45(Suppl 2) :152A (abstract 554)). Based
`on their insulinotropic activities, the use of exendin-3 and
`exendin-4 for the treatment of diabetes mellitus and the
`prevention of hyperglycemia has been proposed (Eng, U.S. Patent
`No. 5,424,286).
`The results of an investigation which sho~ed that exendins
`are not the species homolog of mammalian GLP-1 was reported by
`Chen and Drucker who cloned the exendin gene from the Gila
`monster (J. Biol. Chern. 272(7) :4108-15 (1997)). The observation
`that the Gila monster also has separate genes for proglucagons
`(from which GLP-1 is processed), that are more similar to
`mammalian proglucagon than exendin, indicated that exendins are
`not merely species homologs of GLP-1.
`Methods for regulating gastrointestinal motility using
`exendin agonists are described in commonly owned U.S. Patent
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`Application Serial No. 08/908,867, filed August 8, 1997 entitled
`"Methods ~or Regulating Gastrointestinal Motility," which
`application is a continuation-in-part of U.S. Patent Application
`Serial No. 08/694,954, filed August 8, 1996.
`Methods for reducing food intake using exendin. agonists are
`described in commonly owned U.S. Patent Application Serial No.
`09/003,869, filed January 7, 1998, entitled "Use of Exendin and
`Agonists Thereof for the Reduction of Food Intake," which claims
`the benefit of u.s. Provisional Application Nos. 60/034,905 filed
`January 7, 1997, 60/055,404 filed August 7, 1997, 60/065,442
`filed November 14, 1997 and 60/066,029 filed November 14, 1997.
`Novel exendin agonist compounds are described in commonly
`owned PCT Application Serial No. PCT/US98/16387 filed August 6,
`1998, entitled "Novel Exendin Agonist Compounds," which claims
`the benefit of U.S. Patent Application Serial No. 60/055,404,
`filed August 8, 1997.
`Other novel exendin agonists are described in commonly owned
`PCT Application Serial No. PCT/US98/24210, filed November 13,
`1998, entitled "Novel Exendin Agonist Compounds," which claims
`the benefit of U.S. Provisional Application No. 60/065,442 filed
`November 14, 1.997.
`Still other novel exendin agonists are described in commonly
`owned PCT Application Serial No. PCT/US98/24273, filed November
`13, 1998, entitled "Novel Exendin Agonist Compounds," which
`claims the benefit of U.S. Provisional Application No. 60/066,029
`filed November 14, 1997.
`Other recent advances in exendin related technology are
`described in U.S. Provisional Patent Application Serial No.
`60/075,122, filed February 13, 1998, entitled "Inotropic and
`30 Diuretic Effects of Exendin and GLP-1" and in U.S. Provisional
`Patent Application Serial No. 60/116,380, filed January 14, 1998,
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`entitled "Novel Exendin Agonist Formulations and Methods of
`Administration Thereof".
`Polyethylene glycol (PEG) modification of therapeutic
`peptides and proteins may yield both advantages and
`5 disadvantages. While PEG modification may lead to improved
`circulation time, reduced antigenicity arid immunogenicity,
`improved solubility, resistance to proteqlysis, improved
`bioavailability, reduced toxicity, improved stability, and easier
`formulation of peptides (See, Francis et al .. , International
`Journal of Hematology, 68:1-18, 1998) problems with PEGylation in
`most cases is substantial reduction in bioactivity.
`In
`Id.
`addition, most methods involve use of linkers that have several
`types of adverse effects including immunogenicity, instability;
`toxicity, and reactivity.
`Id.
`Modified exendins and exendin agonists and related
`formulations, dosage formulations, and methods that solve these
`.problems and that are useful in the delivery .of therapeutically
`effective amounts of exendins and exendin agonists are described
`and claimed herein.
`The contents of the above-identified articles, patents, and
`patent applications, and all other documents mentioned or cited
`herein, are hereby incorporated by reference in their entirety.
`The inventors reserve the right to physically incorporate into
`this application any and all materials and information from any
`such articles, patents, patent applications, or other documents
`mentioned or cited herein.
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`SUMMARY OF THE INVENTION
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`The present invention relates to novel modified exendins and
`exendin agonists having an exendin or exendiri agonist linked to
`one or more molecular weight increasing compounds, of which
`polyethylene glycol polymers (or other molecular weight
`increasing agents), and related products and methods. Such
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`products and methods that are useful for many applications,
`including, for example, in the treatment of diabetes, including
`Type 1 and 2 diabetes, gestational diabetes (see U.S. patent
`application.serial no. 09/323,867, entitled, "Use of Exendins and
`5 Agonists Thereof For The Treatment of Gestational Diabetes
`Mellitus," filed June 1, 1999), in the treatment of disorders
`which would be benefited by agents which modulate plasma glucose
`levels, in the treatment of disorders which would be benefited by
`the administration of agents useful in modulating the rate of
`10 gastric emptying or food intake, including obesity, eating
`disorders, and insulin-resis.tance syndrome, and to modulate
`triglyceride levels and to treat subjects suffering from
`dyslipidemia (i.e., increased LDL cholesterol, increased VLDL
`cholesterol, and/or decreased HDL cholesterol) (see U.S.
`provisonal patent application serial no. 60/175,365, entitled,
`"Use of Exendins and Agonists Thereof for Modulation of
`Triglyceride Levels and Treatment of Dyslipidem~a," filed January
`10, 2000). The methods are also useful for lowering plasma lipid
`levels, reducing cardiac risk, reducing the appetite, and
`reducing the weight of subjects. Still other embodiments concern
`methods for suppressing glucagon secretion (see U.S. provisonal
`patent application serial no. 60/132,017, entitled, "Methods for
`Glucagon· Suppression," filed April 30, 1999, which is commonly
`owned) . Pharmaceutical compositions for use in the methods of
`the invention are also disclosed.
`The present invention is related to the surprising discovery
`that exendin is cleared from the plasma almost entirely by renal
`filtration, and not primarily by proteolytic degradation, as
`occurs for many other biologically active peptides, for example,
`30 GLP-1. This surprising discovery supports the determination that
`PEGylation or other modification of exendin or exendin agonists
`to increase molecular size, will have pharmaceutical benefit.
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`Thus, the present invention provides a modified exendin or
`exendin agonist having an exendin or exendin agonist linked to
`one or more polyethylene glycol polymers or other molecular
`weight increasing compounds. A "molecular weight increasing
`compound" is one that can be conjugated to an exendin or exendin
`agonist and thereby increase the molecular weight of the
`resulting conjugate. Representative examples of molecular weight
`increasing compounds, in addition to PEG, are polyamine acids
`(e.g., poly-lysine, poly-glutamic acid, and poly-aspartic acid;
`see Gombotz, et al. (1995), Bioconjugate Chern., vol. 6: 332-351;
`Hudecz, et al. (1992), Bioconjugate Chern., vol. 3, 49-57;
`Tsukada, et al. (1984), J. Natl. Cancer Inst., vol 73,: 721-729;
`Pratesi, et al. (1985), Br. J. Cancer, vol. 52: 841-848),
`particularly those of the L conformation, pharmacologically
`ina~tive proteins (e.g., albumini see Gombotz, et al. (1995) and
`the references cited therein), gelatin (see Gombotz, et al.
`(1995) and the references cited therein), succinyl-gelatin (see
`Gombotz, et al. (1995) and the references cited therein),
`(hydroxypropyl) -methacrylamide (see Gombotz, et al. (1995) .and
`the references cited therein), a fatty acid, a olysaccaride, a
`lipid amino acid, and dextran.
`In preferred embodiments, the modified exendin or exendin
`agonist has a molecular weight that is greater than the molecular
`weight of the exendin or exendin agonist (preferably about 10%~
`50% or 90% greater), the modified exendin or exendin agonist has
`a negative charge that is greater than the negative charge of the
`exendin or exendin agonist (preferably about 10%, 50% or 90%
`greater), the modified exendin or exendin agonist has a kidney
`clearance that is less than the kidney clearance of the exendin
`30 or exendin agonist (preferably about 10%, 50% or 90% less), the
`modified exendin or exendin agonist has a half-life that is
`greater than the half-life of the exendin or exendin agonist
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`(preferably about 10%, SO% or 90% greater), the modified exendin
`or exendin agonist has a immunogenicity/antigenicity that is less
`than the immunogenicity/antigenicity of the exendin or exendin
`agonist, the modified exendin or exendin agonist has a solubility
`that is greater than the solubility of the exendin or exendin
`agonist (preferably about 10%, SO% or 90% greater), the modified
`exendin or exendin agonist has a proteolysis rate that is less
`than the proteolysis rate of the exendin or exendin agonist
`(preferably about 10%, SO% or 90% less), the modified exendin or
`exendin agonist has a toxicity that is less than the toxicity of
`the exendin or exendin agonist, the modified exendin or exendin
`agonist has a stability that is greater than the stability of the
`exendin or exendin agonist, and/or the modified exendin or
`exendin agonist has a permeability/biological function that is
`greater or less than the permeability/biological function of the
`exendin or exendin agonist (preferably about 10%, SO% or 90%
`greater or less) .
`The exendin or exendin agonist may be linked to one, two or
`three polyethylene glycol polymers or other molecular weight
`increasing·agents. The polyethylene glycol polymers (or other
`molecular weight increasing agents) may preferably have molecular
`weights between SOO and 20,000.
`In a preferred embodiment, the
`modified exendin or exendin agonist is one of compounds 201-230,
`more preferably one of compounds 209, 210 and 213, or one of
`compounds 201 and 202, or one of compounds 216 and 217 (See
`Example 4 below) .
`The polyethylene glycol polymers (or other molecular weight
`increasing agents) are preferably linked to an amino, carboxyl,
`or thio group, and may be linked by N or C termini of side chains
`of lysine, aspartic acid, glutamic acid, or cysteine, or
`alternatively, the polyethylene glycol polymers or other
`molecular weight increasing agents may be linked with diamine and
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`dicarboxylic groups. The exendin or exendin agonist is
`preferably linked to the polyethylene glycol polymers or other
`molecular weight increasing agents through an epsilon amino group
`on a lysine amino acid of the exendin or exendin agonist.
`The present invention also features a method of making a
`modified exendin or exendin agonist. The method involves linking
`one or more polyethylene glycol polymers or other molecular
`In
`weight increasing agents to an exendin or exendin agonist.
`preferred embodiments, the linking is performed by solid-phase
`synthesis.
`The present invention also provides a method of treating a
`disease benefited by administration of an exendin or exendin
`agonist. The method involves providing a modified exendin or
`exendin agonist of the invention to a patient having such a
`disease and thereby treating the disease. Exemplary diseases
`include postprandial dumping syndrome, postprandial
`hyperglycemia, impaired glucose tolerance, a condition or
`disorder which can be alleviated by reducing food intake,
`obesity, an eating disorder, insulin-resistance syndrome,
`diabetes mellitus, and a hyperglycemic condition.
`In a preferred
`embodiment, the postprandial hyperglycemia is a consequence of
`Type 2 diabetes mellitus.
`In other preferred embodiments, the
`postprandial hyperglycemia is a consequence of Type 1 diabetes
`mellitus or impaired glucose tolerance.
`Also featured in the present invention is a pharmaceutical
`composition. The composition contains a modified exendin or
`exendin agonist and a pharmaceutically acceptable carrier.
`The invention also provides a kit. The kit contains a
`modified exendin or exendin agonist and instructions and/or
`packaging for use. The kit may also include a document
`indicating that the kit, its components, or the methods of using
`them, has received regulatory approval.
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`The present invention also provides a method of beneficially
`regulating gastro-intestinal motility in a subject. The method
`involves administering to the subject a therapeutically effective
`amount of a modified exendin or exendin agonist of the present
`invention.
`Also featured are methods of treatment for ingestion of a
`toxin. The methods involve:
`(a) administering an amount of a
`modified exendin or exendin agonist of the present invention
`effective to prevent or reduce the passage of stomach contents to
`the intestines; and (b) aspirating the contents of the stomach.
`The invention also provides methods for reducing the
`appetite or weight, or lowering plasma lipids, of a subject, as
`well as methods for treating gestational diabetes. The invention
`also provides methods for reducing the appetite or weight, or
`lowering plasma lipids, of a subject, as well as methods for
`treating gestational diabetes. Additional methods include
`modulating triglyceride levels, and treating subjects suffering
`from dyslipidemia, as well as suppressing glucagon levels. These
`and other methods of the invention involve administering to the
`subject a therapeutically effective amount of a modified exendin
`or exendin agonist of the present invention.
`Modified exendins and exendin agonists are useful, for
`example, as inhibitors of gastric emptying for the treatment of,
`for example, diabetes mellitus, and obesity. Thus, the present
`invention is also directed to novel methods for reducing gastric
`motility and slowing gastric emptying. The methods involve the
`administration of a modified exendin or exendin agonist, for
`example one or more PEG polymers linked to exendin-3 [SEQ ID NO.
`1], exendin-4 [SEQ ID NO. 2], or other compounds which
`effectively bind to the receptor at which exendins exert their
`action on gastric motility and gastric emptying. These methods
`will be useful in the treatment of, for example, post-prandial
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`hyperglycemia, a complication associated with type 1 (insulin
`dependent) and type 2 (non-insulin dependent) diabetes mellitus,
`as well as gestational diabetes, dyslipidemia, to modulate
`triglyceride levels, and to suppress glucagon secretion.
`By "exendin agonist" is meant a compound which mimics the
`effects of exendins, e.g., on gastric motility and gastric
`emptying (namely, a compound which effectively binds to the
`receptor at which exendins exert their action on gastric motility
`and gastric emptying, preferably an analog or derivative of an
`exendin) or a compound, e.g., that mimics the effects of exendin
`on the reduction of food intake by binding to the receptor or
`receptors where exendin causes this effect. Preferred exendin
`agonist compounds include those described in United States Patent
`Application Serial No. 90/003,869, entitled, "Use of Exendin And
`-15 Agonists Thereof For The Reduction of Food Intake", filed January
`7, 1998, (and the priority applications thereto) which enjoys
`common ownership with the present application and which is
`incorporated by this reference into the present application as
`though fully set forth herein. Effects of exendins or exendin
`agonists on reducing food intake can be identified, evaluated, or
`screened for, using the methods described herein, or other
`methods known in the art for determining exendin effects, e.g.,
`on food intake or appetite.
`In another aspect, a therapeutically effective amount of an
`amylin agonist is also administered to the subject.
`In a
`preferred aspect, the amylin agonist is an amylin or an amylin
`agonis~ analog such as 25
`29Pro-human-amylin. The use of amylin
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`agonists to treat post-prandial hyperglycemia, as well as to
`beneficially regulate gastrointestinal motility, is described in
`International Application No. PCT/US94/10225, published March 16,
`1995 which has been incorporated by reference herein.
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`In yet another aspect, a therapeutically effective amount of
`an insulin or insulin analog is also administered, separately or
`together with a modified exendin or exendin agonist, to. the
`subject.
`Preferably, the subject is a vertebrate, more preferably a
`In preferred aspects, the
`mammal, and most preferably a human.
`modified exendin or exendin agonist of the invention is
`In a
`administered parenterally, more preferably by injection.
`most preferred aspect, the injection is a peripheral injection~.
`10 Preferably, about 1 ~g-30 ~g to about 5 mg of the modified
`exendin or exendin agonist of the invention is administered per
`day. More preferably, about 1-30 ~g to about 2mg, or about 1-30
`~g to about 1mg of the modified exendin or exendin agonist of the
`invention is administered per day. Most preferably, about 3 ~g
`to about 500 ~g of the modified exendin or exendin agonist of the
`invention is administered per day.
`Preferred exendins or exendin agonists for modification and
`use include:
`[SEQ ID NO 3: His Gly Glu Gly Thr Phe Thr
`exendin-4 (1-30)
`20 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 4: 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-NH2J ;
`exendin-4 (1-28) amide [SEQ ID NO 5: 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-NH2l;
`14Leu, 25Phe exendin- 4 amide [SEQ 'ID NO 6: 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-NH2];
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`25
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`30
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`14Leu, 25 Phe exendin-4 (1-28) amide [SEQ ID NO 7: 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-NH2J ; and
`14Leu, 22Ala, 25Phe exendin-4 (1-28) amide [SEQ ID NO 8: His Gly
`5 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-NH2J .
`In the methods of the'present invention, the modified
`exendins or exendin agonists may be administered separately or
`together with one or more other compounds and compositions that
`exhibit a long term or short-term satiety action, including, but
`not limited to other compounds and compositions that include an
`amylin agonist, cholecystokinin (CCK), or a leptin (ob protein).
`28
`[25
`29Pro-] -human
`Suitable amyl in agonists include, for example,
`amylin (also known as "pramlintide," and previously referred to
`as "AC-137") as described in "Amyl in Agonist Peptides and Uses
`Therefor," U.S. Patent No. 5,686,511, issued November 11, 1997,
`and salmon calcitonin. The CCK used is preferably CCK
`octapeptide (CCK-8) . Leptin is discussed in, for example,
`Pelleymounter, M.A., et al. Science 269:540-43 (1995); Halaas,
`J.L., et al. Science 269:543-46 (1995); and Campfield, L.A., et
`al. Eur. J. Pharmac. 262:133-41 (1994).
`The invention also provides compositions and methods for
`providing therapeutically effective amounts of the modified
`exendins or exendin agonists of the invention in order to
`increase urine flow in an individual, decrease the amount of
`potassium in the urine of an individual, prevent or alleviate a
`condition or disorder associated with hypervolemia or toxic
`hypervolemia in an individual, induce rapid diuresis, prepare an
`individual for a surgical procedure, increase renal plasma flow
`and glomerular filtration rates, or treat pre-eclampsia or
`eclampsia of pregnancy.
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`Definitions
`In accordance with the present invention and as used herein,
`the following terms are defined to have the following meanings,
`unless explicitly stated otherwise.
`The term "amino acid" refers to natural amino acids,
`unnatural amino acids, and amino acid analogs, all in their D and
`L stereoisomers if their structure allow such stereoisomeric
`forms. Natural amino acids include alanine (Ala), arginine
`(Arg), asparagine (Asn), aspartic acid (Asp), cysteine (Cys),
`glutamine (Gln), glutamic acid (Glu), glycine (Gly), histidine
`(His), isoleucine (Ile), leucine (Leu), Lysine (Lys), methionine
`(Met), phenylalanine (Phe), proline (Pro), serine (Ser),
`threonine (Thr) I
`typtophan (Trp) I
`tyrosine (Tyr) and valine
`(Val) . Unnatural amino acids include, but are not limited to
`azetidinecarboxylic acid, 2-aminoadipic acid, 3-aminoadipic acid,
`beta-alanine, aminopropionic acid, 2-aminobutyric acid, 4-
`aminobutyric acid, 6-aminocaproic acid, 2-aminoheptanoic acid, 2-
`aminoisobutyric acid, 3-aminoisbutyric acid, 2-aminopimelic acid,
`tertiary-butylglycine, 2,4-diaminoisobutyric acid, desmosine,
`2,2'-diaminopimelic acid, 2,3-diaminopropionic acid, N(cid:173)
`ethylglycine, N-ethylasparagine, homoproline, hydroxylysine,
`allo-hydroxylysine, 3-hydroxyproline, 4-hydroxyproline,
`isodesmosine, allo-isoleucine, N-methylalanine, N-methylglycine,
`N-methylisoleucine, N-methylpentylglycine, N-methylvaline,
`'naphthalanine, norvaline, norleucine, ornithine, pentylglycine,
`pipecolic acid and thioproline. Amino acid analogs include the
`natural and unnatural amino acids which are chemically blocked,
`reversibly or irreversibly, or modified on their N-terminal amino
`group or their side chain groups, as for example, methionine
`sulfoxide, methionine sulfone, S-(carboxymethyl)-cysteine, S(cid:173)
`(carboxymethyl)-cysteine sulfoxide and S-(carboxymethyl)-cysteine
`sulfone.
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`The term "amino acid analog" refers to an amino acid wherein
`either the C-terminal carboxy group, the N-terminal amino group
`or side chain functional group has been chemically codified to
`another functional group. For example, aspartic acid-(beta-
`5 methyl ester) is an amino acid analog of aspartic acid; N(cid:173)
`ethylglycine is an amino acid analog of glycine; or alanine
`carboxamide is an amino acid analog of alanine.
`The term "amino acid residue" refers to radicals having the
`structure: (1) -C(O)-R-NH-, wherein R typically is -CH(R')-,
`10 wherein R' is an amino acid side chain, typically H or a carbon
`containing substitutent;
`
`or (2)
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`C(=O}-
`
`N
`I
`
`, wherein p is 1, 2, or 3 representing the azetidinecarboxylic
`acid, proline, or pipecolic acid residues, respectively.
`The term "lower" referred to herein in connection with
`organic radicals such as alkyl groups defines such groups with up
`to and including about 6, preferably up to and including 4 and
`advantageously one or two carbon atoms. Such groups may be
`straight chain or branched chain.
`"Pharmaceutically acceptable salt" includes salts of the
`compounds of the present invention derived from the combination
`of such compounds and an organic or inorganic acid.
`In practice
`.the use of the salt form amounts to use of the base form.
`.The
`compounds of the present invention are useful in both free base
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`and salt form, with both forms being considered as being within
`the scope of the present invention.
`In addition, the following abbreviations stand for the
`following:
`"ACN" or "CH3CN" refers to acetonitrile.
`"Boc", "tBoc" or "Tboc" refers to t-butoxy carbonyl.
`"DCC" refers to N,N'-dicyclohexylcarbodiimide.
`"Fmoc" refers to fluorenylmethoxycarbonyl.
`"HBTU" refers to 2-(1H-benzotriazol-l-yl)-
`1,1,3,3,-tetramethyluronium hexaflurophosphate.
`"HOBt" refers to 1-hydroxybenzotriazole monohydrate.
`"homoP" or hPro" refers to homoproline.
`"MeAla" or "Nme" refers to N-methylalanine.
`"naph" refers to naphthylalanine.
`"pG" or pGly" refers to