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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
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`of, United States provisional patent application serial
`
`no. 60/132,018, filed April 30, 1999, which application is
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`hereby incorporated by reference in its entirity.
`
`FIELD OF THE INVENTION
`
`The present
`
`invention relates to novel modified
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`exendins and exendin agonists having an exendin or exendin
`
`agonist peptide linked to one or more polyethylene glycol
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`5
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`polymers (or other molecular weight increasing agents),
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`nI.-
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`IA...-4Wait4...]!I4.-.
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`and related products and methods that are useful,
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`for
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`example,
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`in the treatment of diabetes,
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`including Type 1
`
`and 2 diabetes,
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`in the treatment of disorders which would
`
`be benefited by agents which modulate plasma glucose
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`20
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`levels, and in the treatment of disorders which would be
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`benefited by the administration of agents useful in
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`modulating glucagon or triglyceride levels, or the rate of
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`gastric emptying or food intake,
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`including obesity, eating
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`disorders, and insulin—resistance syndrome.
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`25
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`BACKGROUND
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`The following description includes information that
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`may be useful
`SD-149563.!
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`in understanding the present
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`invention.
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`It
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`SANOFI-AVENTIS Exhibit 1015 - Page 1
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`is not an admission that any of the information provided
`herein is prior art to the presently claimed invention,
`nor that any of the publications specifically or
`implicitly referenced are prior art to that invention.
`The exendins are peptides that are found in the
`salivary secretions of the Gila monster and the Mexican
`Bearded Lizard, reptiles that are endogenous to Arizona
`and Northern Mexico. Exendin-3 [SEQ. ID. NO. 1] is
`present in the salivary secretions of Heloderma horridum
`(Mexican Beaded Lizard), and exendin-4 [SEQ. ID. NO. 2] is
`present in the salivary secretions of Heloderm suspectum
`(Gila monster) (Eng, J., et al., J. Biol. Chem., 265:20259-
`62, 1990; Eng, J., et al., J. Biol. Chem., 267:7402-05,
`1992). The amino acid sequence of exendin-3 is shown in
`Figure 1. The amino acid sequence of exendin-4 is shown
`in Figure 2. Exendin-4 was first thought to be a
`(potentially toxic) component of the venom.
`It now
`appears that exendin-4 is devoid of toxicity, and that it
`instead is made in salivary glands in the Gila monster.
`The exendins have some sequence similarity to several
`members of the glucagon-like peptide family, with the
`highest homology, 53%, being to GLP-1[7-36]NH2 [SEQ. ID.
`NO. 3]
`(Goke, et 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", has an
`insulinotropic effect, stimulating insulin secretion from
`pancreatic beta-cells; GLP-1 has also been reported to
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`SD-149563.1
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`SANOFI-AVENTIS Exhibit 1015 - Page 2
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`inhibit glucagon secretion from pancreatic a1pha—cells
`
`(flrsov, et al., Diabetes, 42:658-61, 1993; D’Alessio, et
`
`al., J. Clin. Invest., 972133-38, 1996).
`
`GLP—1 has been
`
`reported to inhibit gastric emptying (willms B, et al., J.
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`5 Clin. Endocrinol. Metab. 81 (1): 327-32, 1996; Wettergren
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`A, et al., Dig. Dis. Sci. 38
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`(4): 665-73, 1993), and
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`gastric acid secretion (Schjoldager BT, et al., Dig. Dis.
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`Sci. 34
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`(5): 703-8, 1989; O'Halloran DJ, et al., J.
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`Endocrinol. 126 (1): 169-73, 1990; Wettergren A, et al.,
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`10 Dig. Dis. Sci. 38
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`(4): 665-73, 1993)). GLP—1[7—37], which
`
`has an additional glycine residue at its carboxy terminus,
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`is reported to stimulate insulin secretion in humans
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`(flrsov, et al., Diabetes, 42:658-61, 1993). Other reports
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`relate to the inhibition of glucagon secretion
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`I5
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`(Creutzfeldt WOC, et al., Glucagonostatic actions and
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`reduction of fasting hyperglycemia by exogenous glucagon-
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`like peptide I(7-36) amide in Type 1 diabetic patients,
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`Diabetes Care 1996;19(6):580—6), and a purported role in
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`appetite control
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`(Turton MD, et al., A role for glucagon-
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`20
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`like peptide-1 in the central regulation of feeding,
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`Nature 1996 Jan;379(6560):69—72).A transmembrane G—protein
`
`adenylate-cyclase-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,
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`25
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`Proc. Natl. Acad. Sci. USA 89:8641-45, 1992). GLP-1 has
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`been the focus of significant investigation in recent
`
`years due to its reported action on the amplification of
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`SD-l49563.l
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`SANOFI-AVENTIS Exhibit 1015 - Page 3
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`vllilhIL’?'.Z‘.nr.‘.,,IV"):i‘..nl'.-muIlu-Vll-nix
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`stimulated insulin production (Byrne MM, Goke B. Lessons
`
`from human studies with glucagon—like_peptide~1: Potential
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`of the gut hormone for clinical use.
`
`In:
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`Fehmann HC, Goke
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`B.
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`Insulinotropic Gut Hormone Glucagon-Like Peptide 1.
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`Basel, Switzerland: Karger, 1997:2l9—33).
`
`GLP-1 has also been reported to restore islet glucose
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`sensitivity in aging rats,
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`restoring their glucose
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`tolerance to that of younger rats (Egan JM, et al.,
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`Diabetologia 1997 Jun;40(Suppl l):Al30). However,
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`the
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`short duration of biological action of GLP-1 in vivo is
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`one feature of the peptide that has hampered its
`
`development as a therapeutic agent. Various methods have
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`been tried to prolong the half~life of GLP—l 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)).
`
`Pharmacological studies have led to reports that
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`exendin—4 can act at GLP~l receptors 33 vitro on certain
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`insulin-secreting cells, at dispersed acinar cells from
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`guinea pig pancreas, and at parietal cells from stomach;
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`the peptide is also reported to stimulate somatostatin
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`release and inhibit gastrin release in isolated stomachs
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`(Goke, et al., J. Biol. Chem. 268:l9650-S5, 1993; Schepp,
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`SD-149563. I
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`et al., Eur. J. Pharmacol., 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, 4l:l49-56, 1992: Raufman,
`
`et al., J. Biol. Chem. 267:2l432-37, 1992; Singh, et a1.,
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`Regul. Pept. 53:47-59, 1994). Exendin-4 has a
`
`significantly longer duration of action than GLP-1.
`
`For
`
`example,
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`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.
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`Prolonged effect of exendin—4 on hyperglycemia of db/db
`
`mice, Diabetes 1996 May; 45(Suppl 2):152A (abstract 554)).
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`Based on their insulinotropic activities,
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`the use of
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`exendin-3 and exendin-4 for the treatment of diabetes
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`mellitus and the prevention of hyperglycemia has been
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`proposed (Eng, U.S. Patent No. 5,424,286).
`
`The results of an investigation which showed that
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`exendins are not
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`the species homolog of mammalian GLP-1
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`was reported by Chen and Drucker who cloned the exendin
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`gene from the Gila monster
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`(J. Biol. Chem. 272(7):4l08-15
`
`(1997)).
`
`The observation that the Gila monster also has
`
`separate genes for proglucagons (from which GLP-1 is
`
`processed),
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`that are more similar to mammalian proglucagon
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`than exendin,
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`indicated that exendins are not merely
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`species homologs of GLP—1.
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`Methods for regulating gastrointestinal motility
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`using exendin agonists are described in commonly owned
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`U.S. Patent Application Serial No. 08/908,867, filed
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`August 8, 1997 entitled "Methods for Regulating
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`5 Gastrointestinal Motility," which application is a
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`continuation-in-part of U.S. Patent Application Serial
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`No. 08/694,954, filed August 8, 1996.
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`Methods for reducing food intake using exendin
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`agonists are described in commonly owned U.S. Patent
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`10 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
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`U.S. Provisional Application Nos. 60/034,905 filed January
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`7, 1997 1 60/055,404 filed August 7 1 1997 1 60/065 1 442 filed
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`15 November 14 1 1997 and 60/066,029 filed November 14 1 1997.
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`Novel exendin agonist compounds are described in
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`commonly owned PCT Application Serial No. PCT/US98/16387
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`filed August 6 1 1998, entitled "Novel Exendin Agonist
`
`Compounds," which claims the benefit of U.S. Patent
`
`20 Application Serial No. 60/055 1 404 1 filed August 8, 1997.
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`Other novel exendin agonists are described in
`
`commonly owned PCT Application Serial No. PCT/US98/24210,
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`filed November 13, 1998 1 entitled "Novel Exendin Agonist
`
`Compounds," which claims the benefit of U.S. Provisional
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`25 Application No. 60/065,442 filed November 14, 1997.
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`Still other novel exendin agonists are described in
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`commonly owned PCT Application Serial No. PCT/US98/24273,
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`SD-149563.1
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`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 Diuretic Effects of Exendin and GLP-1" and
`in U.S. Provisional Patent Application Serial
`No. 60/116,380, filed January 14, 1998, entitled "Novel
`10 Exendin Agonist Formulations and Methods of Administration
`Thereof".
`Polyethylene glycol (PEG) modification of therapeutic
`peptides and proteins may yield both advantages and
`disadvantages. While PEG modification may lead to
`improved circulation time, reduced antigenicity and
`immunogenicity, improved solubility, resistance to
`proteolysis, 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 addition,
`Id.
`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
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`SD-149563.1
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`therapeutically effective amounts of exendins and exendin
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`agonists are described and claimed herein.
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`The contents of the above-identified articles,
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`patents, and patent applications, and all other documents
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`mentioned or cited herein, are hereby incorporated by
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`reference in their entirety.
`
`The inventors reserve the
`
`right to physically incorporate into this application any
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`and all materials and information from any such articles,
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`patents, patent applications, or other documents mentioned
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`or cited herein.
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`SUMMARY OF THE INVENTION
`
`The present
`
`invention relates to novel modified
`
`exendins and exendin agonists having an exendin or exendin
`
`agonist linked to one or more molecular weight
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`increasing
`
`compounds, of which polyethylene glycol polymers (or other
`
`molecular weight
`
`increasing agents), and related products
`
`and methods.
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`Such products and methods that are useful
`
`for many applications,
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`including,
`
`for example,
`
`in the
`
`treatment of diabetes,
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`including Type 1 and 2 diabetes,
`
`gestational diabetes (see U.S. patent application serial
`
`no. 09/323,867, entitled, “Use of Exendins and 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
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`disorders which would be benefited by the administration
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`of agents useful
`SD-M95611
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`in modulating the rate of gastric
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`emptying or food intake, including obesity, eating
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`disorders, and insulin-resistance syndrome, and to
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`modulate triglyceride levels and to treat subjects
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`suffering from dyslipidemia (i.e., increased LDL
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`5
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`cholesterol, increased VLDL cholesterol, and/or decreased
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`HDL cholesterol) (see U.S. provisonal patent application
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`serial no. 60/175,365, entitled, "Use of Exendins and
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`Agonists Thereof for Modulation of Triglyceride Levels and
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`Treatment of Dyslipidemia," filed January 10, 2000). The
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`10 methods are also useful for lowering plasma lipid levels,
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`reducing cardiac risk, reducing the appetite, and reducing
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`the weight of subjects. Still other embodiments concern
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`methods for suppressing glucagon secretion (see U.S.
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`provisonal patent application serial no. 60/132,017,
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`15
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`entitled, "Methods for Glucagon Suppression," filed April
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`30, 1999, which is commonly owned). Pharmaceutical
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`compositions for use in the methods of the invention are
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`also disclosed.
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`The present invention is related to the surprising
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`20
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`discovery that exendin is cleared from the plasma almost
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`entirely by renal filtration, and not primarily by
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`proteolytic degradation, as occurs for many other
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`biologically active peptides, for example, GLP-1. This
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`surprising discovery supports the determination that
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`25
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`PEGylation or other modification of exendin or exendin
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`agonists to increase molecular size, will have
`
`pharmaceutical benefit.
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`SD-149563.1
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`5
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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.,
`10 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 inactive proteins (e.g.,
`albumin; see Gombotz, et al. (1995) and the references
`cited therein), gelatin (see Gombotz, et al. (1995) and
`the references cited therein), succinyl-gelatin (see
`20 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
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`SD·I49563.1
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`modified exendin or exendin agonist has a negative charge
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`that is greater than the negative charge of the exendin or
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`exendin agonist (preferably about 10%, 50% or 90%
`
`greater), the modified exendin or exendin agonist has a
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`5
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`kidney clearance that is less than the kidney clearance of
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`the exendin 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 (preferably about 10%, 50% or
`
`10
`
`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
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`15
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`exendin or exendin agonist (preferably about 10%, 50% 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%,
`
`50% or 90% less), the modified exendin or exendin agonist
`
`20
`
`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
`
`25
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`permeability/biological function that is greater or less
`
`than the permeability/biological function of the exendin
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`SD-149563.1
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`or exendin agonist (preferably about 10%, 50% or 90%
`
`greater or less).
`
`The exendin or exendin agonist may be linked to one,
`
`two or three polyethylene glycol polymers or other
`
`5 molecular weight increasing agents. The polyethylene
`
`glycol polymers (or other molecular weight increasing
`
`agents) may preferably have molecular weights between 500
`
`and 20,000.
`
`In a preferred embodiment, the modified
`
`exendin or exendin agonist is one of compounds 201-230,
`
`10 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
`
`15
`
`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
`
`20
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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.
`
`25·
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`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
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`SD-149563.1
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`or other molecular weight increasing agents to an exendin
`
`or exendin agonist.
`
`In 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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`SD-149563.!
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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
`
`5
`
`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
`
`10
`
`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
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`subject, as well as methods for treating gestational
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`15
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`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
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`20
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`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.
`
`25
`
`Modified exendins and exendin agonists are useful,
`
`for example, as inhibitors of gastric emptying for the
`
`treatment of, for example, diabetes mellitus, and obesity.
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`SD-149563.1
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`SANOFI-AVENTIS Exhibit 1015 - Page 14
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`Thus, the present invention is also directed to novel
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`methods for reducing gastric motility and slowing gastric
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`emptying. The methods involve the administration of a
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`modified exendin or exendin agonist, for example one or
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`5 more PEG polymers linked to exendin-3 [SEQ ID NO. 1],
`
`exendin-4 [SEQ ID NO. 2], or other compounds which
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`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
`
`10
`
`example, pos
`
`1 hyperglycemia, a complication
`
`associated with type 1 (insulin dependent) and type 2
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`(non-insulin dependent) diabetes mellitus, as well as
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`gestational diabetes, dyslipidemia, to modulate
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`triglyceride levels, and to suppress glucagon secretion.
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`15
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`By "exendin agonistn is meant a compound which mimics
`
`the effects of exendins, e.g., on gastric motility and
`
`gastric emptying (namely, a compound which effectively
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`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.,
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`20
`
`that mimics the effects of exendin on the reduction of
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`food intake by binding to the receptor or receptors where
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`exendin causes this effect. Preferred exendin agonist
`
`compounds include those described in United States Patent
`
`25 Application Serial No. 90/003,869, entitled, "Use of
`
`Exendin And Agonists Thereof For The Reduction of Food
`
`Intaken, filed January 7, 1998,
`
`(and the priority
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`SD-149563.1
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`SANOFI-AVENTIS Exhibit 1015 - Page 15
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`applications thereto) which enjoys common ownership with
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`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
`
`5
`
`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
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`10
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`of an amylin agonist is also administered to the subject.
`
`In a preferred aspect, the amylin agonist is an amylin or
`28
`an amylin agonist analog such as 25
`29 Pro-human-amylin.
`
`'
`
`'
`
`The use of amylin agonists to treat post-prandial
`
`hyperglycemia, as well as to beneficially regulate
`
`15 gastrointestinal motility, is described in International
`
`Application No. PCT/US94/10225, published March 16, 1995
`
`which has been incorporated by reference herein.
`
`In yet another aspect, a therapeutically effective
`
`amount of an insulin or insulin analog is also
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`20
`
`administered, separately or together with a modified
`
`exendin or exendin agonist, to the subject.
`
`Preferably, the subject is a vertebrate, more
`
`preferably a mammal, and most preferably a human.
`
`In
`
`preferred aspects, the modified exendin or exendin agonist
`
`25
`
`of the invention is administered parenterally, more
`
`preferably by injection.
`
`In a most preferred aspect, the
`
`injection is a peripheral injection. Preferably, about 1
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`SD-149563.1
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`SANOFI-AVENTIS Exhibit 1015 - Page 16
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`~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
`
`5
`
`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:
`
`10
`
`exendin-4 (1-30)
`
`[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];
`
`exendin-4 (1 30) amide [SEQ ID NO 5: His Gly Glu Gly
`
`Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu Glu Ala
`
`15 Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly-NH 2 ];
`
`exendin-4 (1-28) amide [SEQ ID NO 6: 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 7: His Gly Glu
`
`20 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 ];
`14Leu, 25 Phe exendin-4 (1-28) amide [SEQ ID NO 8: His
`
`Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu
`
`25 Glu Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn-NH 2 ];
`and
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`SD- I 49563.1
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`SANOFI-AVENTIS Exhibit 1015 - Page 17
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`14 Leu, 22Ala, 25 Phe exendin-4 (1-28) amide [SEQ ID NO 9:
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`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(cid:173)
`
`NH2] .
`
`5
`
`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
`
`10
`
`compounds and compositions that include an amylin agonist,
`
`cholecystokinin (CCK), or a leptin (ob protein). Suitable
`28
`[ 25
`29 Pro-] -human
`
`amylin agonists include, for example,
`
`'
`
`'
`
`amylin (also known as "pramlintide," and previously
`
`referred to as "AC-137") as described in "Amylin Agonist
`
`15
`
`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.
`
`20
`
`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
`
`25
`
`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
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`SD-149563.1
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`19
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`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
`
`5
`
`eclampsia of pregnancy.
`
`Definitions
`
`In accordance with the present invention and as used
`
`herein, the following terms are defined to have the
`
`10
`
`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
`
`15
`
`(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),
`
`20
`
`threonine (Thr), typtophan (Trp), 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
`
`25
`
`acid, 2-aminoheptanoic acid, 2-aminoisobutyric acid, 3-
`
`aminoisbutyric acid, 2-aminopimelic acid, tertiary(cid:173)
`
`butylglycine, 2,4-diaminoisobutyric acid, desmosine, 2,2'-
`
`SD-149563.1
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`SANOFI-AVENTIS Exhibit 1015 - Page 19
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`diaminopimelic acid, 2,3-diaminopropionic acid, N(cid:173)
`
`ethylglycine, N-ethylasparagine, homoproline,
`
`hydroxylysine, allo-hydroxylysine, 3-hydroxyproline, 4-
`
`hydroxyproline, isodesmosine, allo-isoleucine, N-
`
`5 methylalanine, N-methylglycine, N-methylisoleucine, N(cid:173)
`
`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
`
`10
`
`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(cid:173)
`
`(carboxymethyl)-cysteine, S-(carboxymethyl)-cysteine
`
`sulfoxide and S-(carboxymethyl)-cysteine sulfone.
`
`15
`
`The term "amino acid analogu refers to an amino acid
`
`wherein either the C-terminal carboxy group, the N(cid:173)
`
`terminal amino group or side chain functional group has
`
`been chemically codified to another functional group. For
`
`example, aspartic acid-(beta-methyl ester) is an amino
`
`20
`
`acid analog of aspartic acid; N-ethylglycine is·an amino
`
`acid analog of glycine; or alanine carboxamide is an amino
`
`acid analog of alanine.
`
`The term "amino acid residueu refers to radicals
`
`having the structure:
`
`(1) -C(O)-R-NH-, wherein R typically
`
`25
`
`is -CH(R' )-, wherein R' is an amino acid side chain,
`
`typically H or a carbon containing substitutent;
`
`SD-149563.1
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`SANOFI-AVENTIS Exhibit 1015 - Page 20
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`or (2)
`
`(CH)p
`
`C(~O)
`
`o--
`
`N
`
`I
`
`5
`
`, wherein p is 1, 2, or 3 representing the
`
`azetidinecarboxylic acid, proline, or pipecolic acid
`
`residues,
`
`ively.
`
`The term "lower" referred to herein in connection
`
`10 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.
`
`15
`
`"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
`
`20
`
`a