This Application is a continuation of U.S. Ser. No. 13/464, 045 which is a divisional application of U.S. Ser. No. 12/092,433, now U.S. Pat. No. 8,202,837, which is a Â§371 of International Application No. PCT/US2006/060508, filed 3 Nov. 2006, which claims priority of U.S. Provisional Application No. 60/733,920, filed 4 Nov. 2005 and U.S. Provisional Application No. 60/742,600, filed 6 Dec. 2005.

BACKGROUND

Hypoglycemic agents may be used in the treatment of both Type I and Type II diabetes to lower glucose concentration in blood. Insulinotropic peptides have been implicated as possible therapeutic agents for the treatment of diabetes. Insulinotropic peptides include incretin hormones such as, but are not limited to, gastric inhibitory peptide (GIP) and glucagon like peptide-1 (GLP-1) as well as fragments, variants, and conjugates thereof. Insulinotropic peptides also include exendin 3 and exendin 4. GLP-1 is a 30 amino acid long incretin hormone secreted by the L-cells in the intestine. GLP-1 has been shown to stimulate insulin secretion in a physiological and glucose-dependent manner, decrease glucagon secretion, inhibit gastric emptying, decrease appetite, and stimulate proliferation of Î²-cells.

Insulin and insulinotrpoic peptides may be administered via subcutaneous injection, such as with a needle containing device, for example, a pen injector, and/or syringe. Patients may need to inject several times a day to control blood glucose, which can be burdensome as well as painful. Thus, there is a need for methods of administering hypoglycemic agents less frequently and by methods that will minimize such burdensome regimens as well as site injection pain.

SUMMARY OF THE INVENTION

In one embodiment of the present invention, methods are provided for administering a GLP-1 agonist composition comprising at least one polypeptide to a patient in need thereof, comprising the step of injecting the GLP-1 agonist composition via an injection device comprising a tube having a gauge of about 28 or greater, wherein said polypeptide is administered no more than once daily. The polypeptide may be, but is not limited to, GLP-1 or a fragment, variant, and/or conjugate thereof. Certain embodiments of a GLP-1 or a fragment, variant, or conjugate thereof comprise human serum albumin. Human serum albumin, variants and/or fragments thereof, may be conjugated to a GLP-1 or fragment or variant thereof Human serum albumin may be conjugated through a chemical linker, including but not limited to naturally occurring or engineered disulfide bonds, or by genetic fusion to GLP-1, or a fragment or variant thereof

In another aspect of the present invention, methods are provided for treating or preventing a disease in a mammal in need thereof comprising administering a composition comprising at least one polypeptide having GLP-1 agonist activity once weekly, wherein the composition comprises about 0.25 mg to about 104 mg of at least one polypeptide having GLP-1 activity. The mammal may suffer from one or more of the following diseases: Type 1 diabetes, Type II diabetes, obesity and hyperglycemia.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: A device for delivery of lyophilized hypoglycemic agent and/or GLP-1 agonist

FIG. 2: SEQ ID NO:1 which comprises two tandemly oriented GLP-1(7-36)(A8G) fused to the N-Terminus of human serum albumin.

DEFINITIONS

â€œGLP-1 agonist compositionâ€ as used herein means any composition capable of stimulating the secretion of insulin, including, but not limited to an incretin hormone.

â€œIncretin hormoneâ€ as used herein means any hormone that potentiates insulin secretion. One example of an incretin hormone is GLP-1. GLP-1 is an incretin secreted by intestinal L cells in response to ingestion of food. In a healthy individual, GLP-1 plays an important role regulating post-prandial blood glucose levels by stimulating glucose-dependent insulin secretion by the pancreas resulting in increased glucose absorption in the periphery. GLP-1 also suppresses glucagon secretion, leading to reduced hepatic glucose output. In addition, GLP-1 delays gastric emptying time and slows small bowel motility delaying food absorption. GLP-1 promotes continued beta cell competence by stimulating transcription of genes involved in glucose dependent insulin secretion and by promoting beta-cell neogenesis (Meier, et al. â€œGlucagon-Like Peptide 1 and Gastric Inhibitory Polypeptide Potential Applications in Type 2 Diabetes Mellitusâ€ Biodrugs 2003; 17 (2): 93-102). â€œGLP-1 activityâ€ as used herein means one or more of the activities of naturally occurring human GLP-1, including but not limited to, stimulating glucose-dependent insulin secretion, suppressing glucagon secretion, delaying gastric emptying, and promoting beta cell competence and neogenesis.

An â€œincretin mimeticâ€ as used herein is a compound capable of potentiating insulin secretion. An incretin mimetic may be capable of stimulating insulin secretion, increasing beta cell neogenesis, inhibiting beta cell apoptosis, inhibiting glucagon secretion, delaying gastric emptying and inducing satiety in a mammal. An incretin mimetic may include, but is not limited to, any polypeptide which has GLP-1 activity, including but not limited to, exendin 3 and exendin 4, including any fragments and/or variants and/or conjugates thereof

â€œHypoglycemic agentâ€ as used herein means any compound or composition comprising a compound capable of reducing blood glucose. A hypoglycemic agent may include, but is not limited to, any GLP-1 agonist including incretin hormones or incretin mimetics, GLP-1 or fragment, variant and/or conjugate thereof. Other hypoglycemic agents include, but are not limited to, drugs that increase insulin secretion (e.g., sulfonylureas (SU) and meglitinides), increase glucose utilization (e.g., glitazones), reduce hepatic glucose production (e.g., metformin), and delay glucose absorption (e.g., a-glucosidase inhibitors).

â€œPolypeptideâ€ refers to any peptide or protein comprising two or more amino acids joined to each other by peptide bonds or modified peptide bonds, i.e., peptide isosteres. â€œPolypeptideâ€ refers to both short chains, commonly referred to as peptides, oligopeptides or oligomers, and to longer chains, generally referred to as proteins. Polypeptides may contain amino acids other than the 20 gene-encoded amino acids. â€œPolypeptidesâ€ include amino acid sequences modified either by natural processes, such as posttranslational processing, or by chemical modification techniques that are well known in the art. Such modifications are well described in basic texts and in more detailed monographs, as well as in a voluminous research literature. Modifications can occur anywhere in a polypeptide, including the peptide backbone, the amino acid side-chains and the amino or carboxyl termini. It will be appreciated that the same type of modification may be present in the same or varying degrees at several sites in a given polypeptide. Also, a given polypeptide may contain many types of modifications. Polypeptides may be branched as a result of ubiquitination, and they may be cyclic, with or without branching. Cyclic, branched and branched cyclic polypeptides may result from posttranslation natural processes or may be made by synthetic methods. Modifications include acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteine, formation of pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA mediated addition of amino acids to proteins such as arginylation, and ubiquitination. See, for instance, PROTEINSâ€”STRUCTURE AND MOLECULAR PROPERTIES, 2nd Ed., T. E. Creighton, W. H. Freeman and Company, New York, 1993 and Wold, F., Posttranslational Protein Modifications: Perspectives and Prospects, pgs. 1-12 in POSTTRANSLATIONAL COVALENT MODIFICATION OF PROTEINS, B. C. Johnson, Ed., Academic Press, New York, 1983; Seifter, et al., â€œAnalysis for protein modifications and nonprotein cofactorsâ€ , Meth. Enzymol. (1990) 182:626-646 and Rattan, et al., â€œProtein Synthesis: Posttranslational Modifications and Agingâ€ , Ann NY Acad Sci (1992) 663:48-62.

â€œVariantâ€ as the term is used herein, is a polynucleotide or polypeptide that differs from a reference polynucleotide or polypeptide respectively, but retains essential properties. A typical variant of a polynucleotide differs in nucleotide sequence from another, reference polynucleotide. Changes in the nucleotide sequence of the variant may or may not alter the amino acid sequence of a polypeptide encoded by the reference polynucleotide. Nucleotide changes may result in amino acid substitutions, additions, deletions, fusions and truncations in the polypeptide encoded by the reference sequence, as discussed below. A typical variant of a polypeptide differs in amino acid sequence from another, reference polypeptide. Generally, differences are limited so that the sequences of the reference polypeptide and the variant are closely similar overall and, in many regions, identical. A variant and reference polypeptide may differ in amino acid sequence by one or more substitutions, additions, deletions in any combination. A substituted or inserted amino acid residue may or may not be one encoded by the genetic code. A variant of a polynucleotide or polypeptide may be a naturally occurring such as an allelic variant, or it may be a variant that is not known to occur naturally. Non-naturally occurring variants of polynucleotides and polypeptides may be made by mutagenesis techniques or by direct synthesis.

As used herein â€œconjugateâ€ or â€œconjugatedâ€ refers to two molecules that are bound to each other. For example, a first polypeptide may be covalently or non-covalently bounded to a second polypeptide. The first polypeptide may be covalently bound by a chemical linker or may be fused genetically to the second polypeptide, wherein the first and second polypeptide share a common polypeptide backbone.

As used herein â€œtandemly orientedâ€ refers to two or more polypeptides that are adjacent to one another as part of the same molecule. They may be linked either covalently or non-covalently. Two or more tandemly oriented polypeptides may form part of the same polypeptide backbone. Tandemly oriented polypeptides may have direct or inverted orientation and/or may be separated by other amino acid sequences.

As used herein â€œfragment,â€ when used in reference to a polypeptide, is a polypeptide having an amino acid sequence that is the same as part but not all of the amino acid sequence of the entire naturally occurring polypeptide. Fragments may be â€œfree-standingâ€ or comprised within a larger polypeptide of which they form a part or region as a single continuous region in a single larger polypeptide. By way of example, a fragment of naturally occurring GLP-1 would include amino acids 7 to 36 of naturally occurring amino acids 1 to 36. Furthermore, fragments of a polypeptide may also be variants of the naturally occurring partial sequence. For instance, a fragment of GLP-1 comprising amino acids 7-30 of naturally occurring GLP-1 may also be a variant having amino acid substitutions within its partial sequence.

As used herein, â€œreduceâ€ or â€œreducingâ€ blood glucose refers to a decrease in the amount of blood glucose observed in the blood of a patient after administration a hypoglycemic agent.

As used herein â€œdisease associated with elevated blood glucoseâ€ include, but are not limited to, Type I and Type II diabetes and hyperglycemia.

As used herein â€œco-administrationâ€ or â€œco-administeringâ€ as used herein refers to administration of two or more compounds to the same patient. Co-administration of such compounds may be at about the same time (e.g., within the same hour) or it may be within several hours or days of one another. For example, a first compound may be administered once weekly while a second compound is co-administered daily.

DETAILED DESCRIPTION OF THE INVENTION

In one embodiment of the present invention, methods are provided for administering a GLP-1 agonist composition comprising at least one polypeptide to a patient in need thereof, comprising the step of injecting the GLP-1 agonist composition via an injection device comprising a tube having a gauge of about 28 or greater, wherein said polypeptide is administered no more than once daily. As is understood in the art, examples of needles with gauges of 28 or greater include, but are not limited to, 28, 29 and 30 gauge needles. As is also understood in the art, the higher the needle gauge the smaller the needle aperture. An embodiment of the invention comprises a polypeptide that may be, but is not limited to, GLP-1 or a fragment, variant, or conjugate thereof. A GLP-1 or a fragment, variant, or conjugate thereof may comprise human serum albumin. Human serum albumin may be conjugated to the GLP-1 or fragment or variant thereof. Human serum albumin may be conjugated through a chemical linker or genetically fused to the GLP-1 fragment or variant. Examples of GLP-1, fragments or variants, fused with human serum albumin are provided in the following PCT applications: WO 2003/060071, WO 2003/59934, WO 2005/003296, WO 2005/077042.

A further embodiment of the invention comprises one, two, three, four, five, or more tandemly oriented molecules of GLP-1 fused to the N- or C-terminus of human serum albumin or variant thereof. GLP-1 fragments may include, but are not limited to, molecules of GLP-1 comprising, or alternatively consisting of, amino acids 7 to 36 of GLP-1 (hereinafter designated as â€œGLP-1(7-36)â€ ). Variants of GLP-1 or fragments of GLP-1 may include, but are not limited to, substitutions of an alanine residue analogous to alanine 8 of wild type GLP-1, such alanine being mutated to a glycine (hereinafter designated as â€œA8Gâ€ ) (See for Example, the mutants disclosed in U.S. Pat. No. 5,545,618, herein incorporated by reference in its entirety). Other embodiments have such A8G polypetides fused to the N- or C-terminus of albumin or variant thereof. An example of two tandemly oriented GLP-1(7-36)(A8G) fragments/variants fused to the N-terminus of human serum albumin comprises SEQ ID NO:1, which is presented in FIG. 2.

In another aspect of the present invention, the GLP-1 agonist composition further comprises one or more compounds selected from the group of: peroxisome proliferating activated receptor (PPAR) ligand, thiazolidinedione, metformin, insulin, and sulfonylurea. In another aspect, methods are provided comprising the step of co-administering at least one GLP-1 agonist with one or more compounds selected from the group of: peroxisome proliferating activated receptor (PPAR) ligand, thiazolidinedione, metformin, insulin, and sulfonylurea. The GLP-1 agonist composition may have one or more of these compounds in addition to at least one polypeptide.

In another aspect, the GLP-1 agonist composition is lyophilized. In another aspect of this invention, methods are provided that further comprise admixing said GLP-1 agonist composition with a liquid prior to administration of said GLP-1 agonist composition. In yet another aspect, the GLP-1 agonist composition is in solution form, and may be an aqueous solution.

In yet another aspect of the present invention, at least one polypeptide of a GLP-1 agonist composition may be administered to said patient once weekly, twice weekly, once every two weeks, and/or once monthly. In another aspect, the patient suffers from Type II diabetes. An injection device of the invention may be reusable and/or disposable. In one aspect, an injection device comprises a needle. In another aspect the injection a delivery device of the invention comprises a catheter.

In yet another aspect, GLP-1 agonist composition is administered via subcutaneous injection. In another aspect, the injection may be intramuscular or intravenous, intraperitoneal, intranasal, transmucosal or topical. In another aspect, GLP-1 agonist composition is self-administered, meaning a patient receiving an injection administers a GLP-1 agonist composition to himself or herself. Subcutaneous, injections may be administered, for example, at the abdomen, upper arm, and/or thigh.

In another aspect of the present invention, methods are provided for administering a hypoglycemic agent comprising at least one polypeptide to a patient in need thereof, comprising the step of injecting a hypoglycemic agent via an injection device comprising a tube having a gauge of about 28 or greater, wherein said polypeptide is administered no more than once daily. The needle may have a 29 or 30 gauge. In another aspect, the polypeptide of the invention is not insulin. P In another aspect of the present invention methods are provided for treating or preventing a disease in a mammal in need thereof comprising administering a composition comprising at least one polypeptide having GLP-1 agonist activity once weekly, wherein the composition comprises about 0.010 mg to about 104 mg of at least one polypeptide having GLP-1 activity. The disease may be selected from the group consisting of Type 1 diabetes, Type II diabetes, obesity and hyperglycemia. In one aspect, the disease is Type II diabetes.

In another aspect of the present invention, the polypeptide having GLP-1 activity comprises at least one fragment or variant of human GLP-1 genetically fused with human serum albumin. This fragment or variant of GLP-1 may comprise GLP-1(7-36(A8G)). At least one fragment or variant of GLP-1 may be genetically fused to human serum albumin. In another aspect, the polypeptide having GLP-1 activity comprises at least two GLP-1(7-36(A8G)) tandemly and genetically fused to the human serum albumin. In another aspect, the two GLP-1(7-36(A8G)) are genetically fused at the N-terminus of the human serum albumin. In another aspect, at least one polypeptide having GLP-1 activity comprises SEQ ID No.: 1. At least one polypeptide having GLP-1 activity may be administered at a dose of about 0.25 mg to about 32 mg weekly. Some examples of doses for a once weekly administration of a polypeptide having GLP-1 activity include, but are not limited to, 0.010 mg/week, 0.25 mg/week, 0.5 mg/week, 0.8 mg/week, 1.0 mg/week, 2 mg/week, 3.2 mg/week, 8 mg/week, 12.8 mg/week, 32 mg/week, 51.2 mg/week, and/or 104 mg/week.

EXAMPLES

The following examples illustrate various aspects of this invention. These examples do not limit the scope of this invention which is defined by the appended claims.

Example 1

The following devices may be used to deliver a hypoglycemic and/or a GLP-1 agonist composition:

- 1. STATdoseÂ®, GSK, Liquid Drug Product Reusable Auto-injector;
- 2. Autoject miniÂ®, Owen Mumford, Liquid Drug Product Reusable Auto-injector;
- 3. PenletÂ®, Becton Dickinson, Liquid Drug Product Disposable Auto-injector;
- 4. Tigerlily/SnapdragonÂ®, Owen Mumford, Liquid Drug Product Disposable Auto-injector;
- 5. AutoSafety InjectorÂ®, The Medical House, Liquid Drug Product Disposable Auto-injector;
- 6. Liquid Dry InjectorÂ® Becton Dickinson, Lyophilised Drug Product Disposable Pen Injector; and/or
- 7. Pre-filled syringes.

Example 2

A lyophilized hypoglycemic agent and/or GLP-1 agonist composition may be delivered by a device as shown in FIG. 1. This device system can contain a powder comprising a lyophilized hypoglycemic agent and/or GLP-1 agonist and a liquid. This device system can be held upright to mix a contained powder and liquid. The two halves of an injection pen can be pressed together firmly until the powder completely dissolves. Once dissolved the hypoglycemic agent and/or GLP-1 agonist composition can be administered to the patient.

Example 3

A hypoglycemic agent and/or GLP-1 agonist composition can comprise a polypeptide having GLP-1 activity. A hypoglycemic agent and/or GLP-1 agonist composition comprising a polypeptide having GLP-1 activity can be delivered by subcutaneous injection to a person in need thereof, wherein the agent or composition comprises polypeptide having GLP-1 activity at a dose in the range of about 0.010 mg to about 104 mg once weekly. Some examples of doses for a once weekly administration of a GLP-1 agonist composition comprising a polypeptide having GLP-1 activity include, but are not limited to, 010 mg/week, 0.25 mg/week, 0.5 mg/week, 0.8 mg/week, 1.0 mg/week, 2 mg/week, 3.2 mg/week, 8 mg/week, 12.8 mg/week, 32 mg/week, 51.2 mg/week, and/or 104 mg/week.

Example 4

A lyophilized hypoglycemic agent and/or GLP-1 agonist composition can be reconstituted with water for injection. Examples of excipients that can be included in the composition include, but are not limited to, trehalose dehydrate, mannitol, sodium phosphate (such as dibasic, anhydrous and monobasic, monohydrate), polysorbate 80, sodium hydroxide, phosphoric acid, and water for injection.

All patent applications to which this application claims priority are incorporated by reference herein in their entirety as if each application is specifically and individually being fully set forth.