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Vesiculins

Inactive Publication Date: 2009-02-26
ORAYA THERAPEUTICS +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0038]The invention includes methods for treating and/or preventing, in whole or in part, various diseases, disorders, and conditions, including for example, impaired glucose tolerance; impaired fasting glucose; prediabetes; diabetes and/or its complications, including type 1 and type 2 diabetes and their complications; insulin resistance; Syndrome X; obesity and other weight related disorders; fatty liver disease, including non alcoholic and/or alcoholic fatty liver disease; glucose metabolism diseases and disorders; diseases, disorders or conditions that are treated or treatable with insulin; diseases, disorders or conditions that are treated or treatable with a hypoglycemic agent; diseases, disorders, and conditions characterized at least in part by hyperglycemia; diseases, disorders, and conditions characterized at least in part by hypoinsulinemia and/or diseases, disorders, and conditions characterized at least in part by hyperinsulinemia.
[0039]The invention includes methods for treating a subject having or suspected of having or predisposed to, or at risk for, for example, any diseases, disorders and/or conditions characterized in whole or in part by hyperglycemia, insulin resistance, impaired glucose tolerance, and/or impaired fasting glucose, comprising administering a composition comprising a vesiculin, a vesiculin A chain, a vesiculin B chain, a vesiculin variant, a vesiculin derivative and/or a vesiculin fragment or a salt thereof. Such diseases, disorders and/or conditions include but are not limited to those described or referenced herein. Such compounds may be administered in amounts, for example, that are effective to (1) lower serum glucose, (2) lower blood glucose, (3) lower urine glucose, (4) lower fructosamine, (5) lower glycosylated hemoglobin (HbA1c) levels, (6) lower postprandial glycemia, (7) ameliorate impaired glucose tolerance, (8) ameliorate impaired fasting glucose, and/or (9) lower the rate and/or severity of hypoglycemic events, including severe hypoglycemic events. Such compositions include, for example, formulations for delivery by injection, transdermal patch, inhalation, and other non-oral delivery methods.
[0040]The invention includes methods for regulating glycemia in a subj

Problems solved by technology

In type 2 diabetes these cells progressively lose function, and often fail in the late stages of the disease.
While therapeutic regimens exist to replace insulin and amylin function, diabetic individuals remain prone to complications which are a major threat to both the quality and the quantity of life.
Levels of insulin eventually decrease as a result of the loss of β-cell function and eventual β-cell failure.
Despite decades of research into diabetes and its causes, despite enormous research on β-cells themselves and the proteins they produce, and despite the existence and use of therapies for the treatment of people with type 1 and type 2 diabetes, serious problems remain.

Method used

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Examples

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example 1

[0211]In this example vesiculin was isolated using the following materials, including βTC6-F7 cells. Optiprep™ was obtained from Total Lab Systems, the New Zealand distributors for Axis-Shield products. Acetonitrile used was either HiperSolv grade from BDH or UltimAR grade from Mallinckrodt. Trifluoroacetic acid (Protein Sequencer Grade TFA) was obtained from Applied Biosystems. The 5 μm Jupiter C18 column, 250×2.0 mm was purchased from Phenomenex. Mass spectrometry standards consisted of insulin (RP-HPLC purified from Actrapid® Recombinant Human Insulin [Novo Nordisk Pharmaceuticals]), and synthetic somatostatin (H-2260) purchased from Bachem. The MS matrix used was α-cyano-4-hydroxycinnamic acid (α-CHC) and was obtained from Hewlett Packard. Guanadine HCl, iodoacetic acid, modified RPMI (R1383) and Dulbecco's modified Eagles medium (DMEM; D5523) was purchased from Sigma while fetal bovine serum was sourced from Invitrogen. Dithiothreitol was purchased from Boehringer and sequencin...

example 2

[0216]This Example relates to the identification and characterization of a vesiculin.

[0217]Identification of Granule Proteins—Peptides eluted from RP-HPLC were subjected to N-terminal sequencing on an Applied Biosystems Procise™ 492 protein sequencer using chemicals supplied by the manufacturer (Applied Biosystems).

[0218]Molecular Weight Determination—Molecular weights of purified proteins were determined by MALDI-TOF mass spectrometry (Hewlett Packard G2025A). Samples and external standards (10 μM insulin and somatostatin) were mixed 1:1 with α-CHC (33 mM in 1:2 mixture of acetonitrile:1.33% TFA) and co-crystallised after vacuum drying (Hewlett Packard G2024A Sample prep accessory). Mass / charge ratios (m / z) of unknown proteins were generated by accumulating the data from 50 laser shots (Table 1). In this manner, the presence of C-peptide I and II fragments, amylin, ubiquitin, preptin, an IGF-II-like peptide, insulins I and II, neuropeptide Y, chromogranin A, and calreticulin (FIG. ...

example 3

[0225]This Example describes experiments demonstrating vesiculin bioactivity, here the incorporation of glucose into muscle glycogen. The soleus muscle was dissected from anaesthetized fasted male Wistar rats (200 g±20 g). Each muscle was then teased longitudinally into 3 equal strips with a final radius of approximately 1.5 mm. An average of six muscle strips were transferred into flasks, containing 10 ml of carbogen-saturated nDMEM and peptide (vesiculin, insulin-II, or commercial human IGF-II (GroPep)) at concentrations ranging from 0.71 nM to 237 nM, before the addition of 10 μl of (0.5 μCi) D-[14C(U)]-Glucose. Flasks were then incubated in a shaking water bath for 120 minutes at 30° C. under a stream of carbogen.

[0226]After incubation, the muscle strips were removed and blotted dry before being frozen instantly in liquid nitrogen and freeze-dried for 24 hours. The dried weight of muscle strip was recorded before the muscle strips were solubilised in 250 μl of 60% KOH (70° C. wa...

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Abstract

The inventions relate generally to vesiculin peptides and vesiculin peptide chains, and fragments, variants and derivatives thereof, related compositions and formulations and their preparation and use, nucleic acids encoding such vesiculin peptides and vesiculin peptide chains, and fragments, variants and derivatives thereof and related vectors and host cells, hybridomas and antibodies, and methods for the prevention and treatment of conditions, diseases and disorders that would be improved, eased, or lessened by the administration of a composition of the invention, including but not limited to glucose metabolism diseases.

Description

FIELD OF THE INVENTION[0001]The inventions relate generally to peptides and proteins and related compositions and formulations and their preparation and use, nucleic acids encoding such peptides and proteins and related vectors and host cells, and methods for the prevention and treatment of conditions, diseases and disorders that would be improved, eased, or lessened by the administration of a composition of the invention, including but not limited to glucose metabolism diseases and disorders and diseases and disorders and conditions treated or treatable with insulin and other hypoglycemic agents.BACKGROUND OF THE INVENTION[0002]The following includes information that may be useful in understanding the present inventions. It is not an admission that any of the information provided herein is prior art, or relevant, to the presently described or claimed inventions, or that any publication or document that is specifically or implicitly referenced is prior art.[0003]Insulin-like growth ...

Claims

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Application Information

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IPC IPC(8): A61K38/00C07K14/65C07K14/00C12N15/11C12P21/04A61P43/00C07K1/00C12N15/00C12N5/06C12N1/20
CPCA61K38/00C07K14/461C07K14/46C07K14/47C07K14/465A61P43/00A61P5/48A61P5/50A61P3/10
Inventor BUCHANAN, CHRISTINA MAREECOOPER, GARTH JAMES SMITH
Owner ORAYA THERAPEUTICS
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