U.S. patent application number 12/992060 was filed with the patent office on 2011-10-27 for exendins to lower cholesterol and triglycerides.
This patent application is currently assigned to Amylin Pharmaceuticals, Inc.. Invention is credited to Mark Fineman, Leigh MacConell, Kristin Taylor.
Application Number | 20110263496 12/992060 |
Document ID | / |
Family ID | 41340868 |
Filed Date | 2011-10-27 |
United States Patent
Application |
20110263496 |
Kind Code |
A1 |
Fineman; Mark ; et
al. |
October 27, 2011 |
EXENDINS TO LOWER CHOLESTEROL AND TRIGLYCERIDES
Abstract
Provided herein are pharmaceutical formulations containing
exendins, exendin agonists, or exendin analog agonists that are
administered at therapeutic plasma concentration levels over a
sustained period of time to lower total cholesterol levels; to
lower LDL-cholesterol levels; to lower triglyceride levels; to
treat dyslipidemia; to treat and slow the progression of
atherosclerosis; and to treat, prevent, and reduce the risk of
heart attacks and strokes in patients. In the pharmaceutical
formulations and methods of the invention, the exendin may be
exendin-4, an exendin-4 agonist, or an exendin-4 analog agonist.
The pharmaceutical formulations may be polymer-based pharmaceutical
formulations that may be administered once weekly. An exemplary
pharmaceutical formulation comprises 5% (w/w) of exenatide, about
2% (w/w) of sucrose, and about 93% (w/w) of a
poly(lactide-co-glycolide) polymer, wherein the
poly(lactide-co-glycolide) polymer is in the form of microshperes
encapsulating the exenatide.
Inventors: |
Fineman; Mark; (San Diego,
CA) ; MacConell; Leigh; (San Diego, CA) ;
Taylor; Kristin; (San Diego, CA) |
Assignee: |
Amylin Pharmaceuticals,
Inc.
|
Family ID: |
41340868 |
Appl. No.: |
12/992060 |
Filed: |
May 20, 2009 |
PCT Filed: |
May 20, 2009 |
PCT NO: |
PCT/US09/44727 |
371 Date: |
March 1, 2011 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61054883 |
May 21, 2008 |
|
|
|
Current U.S.
Class: |
514/7.4 |
Current CPC
Class: |
A61K 38/26 20130101;
A61P 9/10 20180101; A61K 38/179 20130101; A61K 9/5031 20130101;
A61K 47/34 20130101; A61K 38/2278 20130101; A61P 3/06 20180101 |
Class at
Publication: |
514/7.4 |
International
Class: |
A61K 38/26 20060101
A61K038/26; A61P 9/10 20060101 A61P009/10; A61P 3/06 20060101
A61P003/06 |
Claims
1-32. (canceled)
33. A method for lowering total cholesterol levels, LDL-cholesterol
levels, or triglyceride levels in a patient in need thereof
comprising: identifying a patient in need of a reduction in total
cholesterol levels, LDL-cholesterol levels, or triglyceride levels;
and administering to the patient a pharmaceutical formulation
comprising at least one exendin, exendin agonist or exendin analog
agonist in an amount sufficient to maintain a steady state
geometric mean plasma concentration of the exendin, exendin agonist
or exendin analog agonist of 145 pg/mL to 700 pg/mL to reduce total
cholesterol levels, LDL-cholesterol levels, or triglyceride levels
in the patient.
34. The method of claim 33, wherein the exendin, exendin agonist,
or exendin analog agonist is (i) exendin-4 or (ii) a peptide having
at least 90% sequence identity to exendin-4.
35. The method of claim 33, wherein the steady state average or
minimum plasma concentration of the exendin, exendin agonist, or
exendin analog agonist at a level of 175 pg/mL to 650 pg/mL.
36. The method of claim 35, wherein the steady state average or
minimum plasma concentration of the exendin, exendin agonist, or
exendin analog agonist at a level of 200 pg/mL to 600 pg/mL.
37. The method of claim 36, wherein the steady state average or
minimum plasma concentration of the exendin, exendin agonist, or
exendin analog agonist at a level of 200 pg/mL to 400 pg/mL.
38. The method of claim 33, wherein the pharmaceutical formulation
further comprises at least one polymer.
39. The method of claim 38, wherein the polymer is
poly(lactide-co-glycolide).
40. The method of claim 33, wherein the pharmaceutical formulation
comprises about 5% (w/w) of exenatide, about 2% (w/w) of sucrose,
and about 93% (w/w) of a poly(lactide-co-glycolide) polymer.
41. The method of claim 33, wherein the average or minimum plasma
concentration of the exendin, exendin agonist, or exendin analog
agonist is at the steady state for about one week to about three
months.
42. The method of claim 33, wherein the dosage of the
pharmaceutical formulation is from about 0.5 mg to about 5 mg, and
wherein the dosage comprises from about 1% to about 20% of the
exendin, exendin agonist, or exendin analog agonist.
43. A method for treating dyslipidemia or atherosclerosis in a
patient in need thereof comprising identifying a patient in need of
treatment for dyslipidemia or atherosclerosis; and administering to
the patient a pharmaceutical formulation comprising at least one
exendin, exendin agonist or exendin analog agonist in an amount
sufficient to maintain a steady state average or minimum plasma
concentration of the exendin, exendin agonist, or exendin analog
agonist at a level of 145 pg/mL to 700 pg/mL for at least one week
to treat dyslipidemia or atherosclerosis in the patient.
44. The method of claim 43, wherein the exendin, exendin agonist,
or exendin analog agonist is (i) exendin-4 or (ii) a peptide having
at least 90% sequence identity to exendin-4.
45. The method of claim 43, wherein the average or minimum plasma
concentration of the exendin, exendin agonist, or exendin analog
agonist is at the steady state for about one week to about three
months.
46. The method of claim 43, wherein the pharmaceutical formulation
further comprises at least one polymer.
47. The method of claim 46, wherein the polymer is
poly(lactide-co-glycolide).
48. The method of claim 43, wherein the pharmaceutical formulation
comprises about 5% (w/w) of exenatide, about 2% (w/w) of sucrose,
and about 93% (w/w) of a poly(lactide-co-glycolide) polymer.
49. The method of claim 43, wherein the dosage of the
pharmaceutical formulation is from about 0.5 mg to about 5 mg, and
wherein the dosage comprises from about 1% to about 20% of the
exendin, exendin agonist, or exendin analog agonist.
50. The method of claim 43, wherein the steady state average or
minimum plasma concentration of the exendin, exendin agonist, or
exendin analog agonist at a level of 175 pg/mL to 650 pg/mL.
51. The method of claim 50, wherein the steady state average or
minimum plasma concentration of the exendin, exendin agonist, or
exendin analog agonist at a level of 200 pg/mL to 600 pg/mL.
52. The method of claim 51, wherein the steady state average or
minimum plasma concentration of the exendin, exendin agonist, or
exendin analog agonist at a level of 200 pg/mL to 400 pg/mL.
Description
FIELD OF THE INVENTION
[0001] This application claims priority to U.S. Application No.
61/054,883 filed May 21, 2008, the disclosure of which is
incorporated herein by reference.
FIELD OF THE INVENTION
[0002] The invention relates to the field of medicine, such as the
use of exendins, exendin agonists, and exendin analog agonists to
lower plasma lipid levels and improve cardiovascular health in
patients.
BACKGROUND OF THE INVENTION
[0003] Research from experimental animals, laboratory
investigations, epidemiology and genetic forms of
hypercholesterolemia indicate that elevated cholesterol is a major
cause of coronary heart disease. The Framingham Heart Study also
established that high blood cholesterol is a risk factor for
coronary heart disease. Results of the Framingham study showed that
the higher the cholesterol level, the greater the risk of suffering
from coronary heart disease. On the other end of the spectrum,
coronary heart disease is generally uncommon at total cholesterol
levels below 150 milligrams per deciliter (mg/dL). A series of more
recent trials of cholesterol lowering using statin drugs has
demonstrated that lowering total cholesterol and low density
lipoprotein (LDL) cholesterol reduces the chances of suffering a
major coronary event, such as myocardial infarction, angina, or
coronary artery procedures (e.g., angioplasty, bypass surgery).
Grundy et al, Circulation, 110:227-239 (2004).
[0004] Total serum cholesterol contains three major classes of
lipoproteins: about 60-70% LDL-cholesterol; about 20-30% high
density lipoprotein (HDL) cholesterol; and about 10-15% very low
density lipoprotein (VLDL) cholesterol. While LDL is the primary
target for cholesterol lowering therapy, HDL and VLDL play a role
in atherosclerosis.
[0005] The American Heart Association endorses the National
Cholesterol Education Program (NCEP) guidelines for the detection
of high cholesterol and the guidelines for healthy fasting
lipoproteins. Total cholesterol levels of less than 200 mg/dL are
desirable, while 240 mg/dL and above are high. HDL cholesterol
levels of 60 mg/dL and above are considered protective against
heart disease, while levels less than 40 mg/dL are a major risk
factor for heart disease. LDL cholesterol levels of 129 mg/dL or
less are near or at optimal, while 160 mg/dL or more are high.
Triglyceride levels of less than 150 mg/dL are normal, while 200
mg/dL and above are high. Third Report of the National Cholesterol
Education Program (NCEP) Expert Panel on Detection, Evaluation and
Treatment of High Blood Cholesterol in Adults (Adult Treatment
Panel III) Final Report, National Institutes of Health, NIH
Publication No. 02-5215 (2002).
[0006] Current treatments to reduce elevated triglycerides, total
cholesterol, and LDL-cholesterol in patients include small molecule
oral monotherapy and combination therapy. The first drug of choice
is generally an HMG CoA reductase inhibitor, i.e., statin.
Exemplary statins include atorvastatin, fluvastatin, lovastatin,
pravastatin, rosuvastatin, and simvastatin. Another class of drugs
is the bile ace sequestrants, such as colesevelam, cholestyramine,
and colestipol. Yet another class of drugs is the fibrates, such as
fenofibrate, clofibrate, and gemfibrozil. Other drugs used to treat
high cholesterol include ezetimibe, nicotinic acid, and probucol.
Several of these drugs are used in combination therapy, such as
ADVICOR.RTM. (lovastatin and niacin by Abbott Laboratories);
CADUET.RTM. (atorvastatin and amlodipine by Pfizer, Inc.); and
VYTORIN.TM. (simvastatin and ezetimibe by Merck and Schering
Plough).
[0007] Exendins are peptides that are found in the saliva of the
Gila monster, a lizard endogenous to Arizona, and the Mexican
Beaded Lizard. Exendin-3 is present in the saliva of Heloderma
horridum, and exendin-4 is present in the saliva of Heloderma
suspectum. Eng et al, J. Biol. Chem., 265:20259-62 (1990); Eng et
al, J. Biol. Chem., 267:7402-05 (1992). Exendins have some amino
acid sequence similarity to several members of the glucagon-like
peptide (GLP) family. For example, exendin-4 as 53% sequence
identity with glucagon-like peptide 1 (GLP-1). Goke et al, J. Biol.
Chem., 268:19650-55 (1993). However, exendin-4 is transcribed from
a distinct gene, not the Gila monster homolog of the mammalian
proglucagon gene from which GLP-1 is expressed. Additionally,
exendin-4 is not an analog of GLP-1. The structure of the synthetic
exendin-4 peptide was not created by sequential modification of the
structure of GLP-1. Nielsen et al, Current Opinion in
Investigational Drugs, 4(4):401-405 (2003); Nielsen et al,
Regulatory Peptides, 117:77-88 (2004).
[0008] Exendin-4 is a potent GLP-1 receptor agonist in vitro. The
peptide also stimulates somatostatin release and inhibits gastrin
release in isolated stomachs. Goke et al, J. Biol. Chem.,
268:19650-55 (1993); Schepp et al, Eur. J. Pharmacol., 69:183-91
(1994); Eissele et al, Life Sci., 55:629-34 (1994). Exendin-3 and
exendin-4 were found to be GLP-1 receptor agonists in stimulating
cAMP production in, and amylase release from, pancreatic acinar
cells. Malhotra et al, Regulatory Peptides, 41:149-56 (1992);
Raufman et al, J. Biol. Chem., 267:21432-37 (1992); Singh et al,
Regulatory Peptides, 53:47-59 (1994). The use of the insulinotropic
activities of exendin-3 and exendin-4 for the treatment of diabetes
mellitus and the prevention of hyperglycemia has been proposed.
U.S. Pat. No. 5,424,286. Synthetic exendin-4, generically known as
exenatide, is commercially available as BYETTA.RTM. (Amylin
Pharmaceuticals, Inc. and Eli Lilly & Company). Sustained
administration of exendins has been proposed. US Publication No.
2004/0053819.
[0009] The need exists for new and improved pharmaceutical
compositions and formulations to reduce total cholesterol levels,
LDL cholesterol levels, and triglyceride levels in patients in need
thereof, to treat dyslipidemia, and atherosclerosis in patients in
need thereof, and to reduce the risk of atherosclerosis, heart
attacks and strokes in patients in need thereof. Pharmaceutical
compositions and formulations that meet the needs of these patients
are described herein.
SUMMARY OF THE INVENTION
[0010] Provided herein are methods for lowering total cholesterol
levels, LDL-cholesterol levels, and triglyceride levels (e.g.,
fasting triglyceride levels) in patients in need thereof by
administering to the patients formulations comprising sufficient
amounts of at least one exendin, exendin agonist, or exendin analog
agonist to maintain steady state average, geometric mean, or
minimum plasma concentrations of the exendins, exendin agonists, or
exendin analog agonists effective to reduce the total cholesterol
levels, LDL-cholesterols levels, or triglyceride levels (e.g.,
fasting triglyceride levels) in the patients.
[0011] Provided herein are methods for treating dyslipidemia and
atherosclerosis in patients in need thereof by administering to the
patients formulations comprising sufficient amounts of at least one
exendin, exendin agonist or exendin analog agonist to maintain
steady state average, geometric mean, or minimum plasma
concentrations of the exendins, exendin agonists, or exendin analog
agonists effective to treat dyslipidemia or atherosclerosis in the
patients.
[0012] Provided herein are methods for preventing, treating, and
reducing the risk of heart attacks and strokes in patients in need
thereof by administering to the patients formulations comprising
sufficient amounts of at least one exendin, exendin agonist, or
exendin analog agonist to maintain steady state average, geometric
mean, or minimum plasma concentrations of the exendins, exendin
agonists, or exendin analog agonists effective to treat prevent,
treat, or reduce the risk of heart attacks and strokes in the
patients.
[0013] Provided herein are the use of exendins, exendin agonists,
and exendin analog agonists in the manufacture of medicaments for
use in lowering total cholesterol levels, LDL-cholesterol levels,
and triglyceride levels (e.g., fasting triglyceride levels). Also
provided herein are the uses of exendins, exendin agonists, and
exendin analog agonists in the manufacture of medicaments in
treating dyslipidemia and atherosclerosis, and in reducing the risk
of heart attacks and strokes.
[0014] Provided herein are methods of treating diabetes; methods of
treating obesity; methods of reducing hemoglobin A.sub.1C; and
methods of reducing postprandial blood glucose in patients in need
thereof by administering to the patients a pharmaceutical
formulation comprising at least one exendin, exendin agonist or
exendin analog agonist in an amount sufficient to maintain a steady
state geometric mean plasma concentration of the exendin, exendin
agonist, or exendin analog agonist at a level of about 145 pg/mL to
about 700 pg/mL. In other embodiments, the steady state geometric
mean plasma concentration of the exendin, exendin agonist, or
exendin analog agonist is about 175 pg/mL to about 650 pg/mL; about
200 pg/mL to about 600 pg/mL; about 200 pg/mL to about 400 pg/mL;
or about 300 pg/mL. In other embodiments, the formulation contains
the exendin, exendin agonist or exendin analog agonist in an amount
of about 0.01 mg to about 1 mg; or about 0.1 mg.
[0015] In one embodiment of the pharmaceutical formulations and
methods described herein, the exendin, exendin agonist or exendin
analog agonist is exendin-4, an exendin-4 agonist or an exendin-4
analog agonist.
[0016] In one embodiment, the pharmaceutical formulations comprise
poly(lactide-co-glycolide) polymeric microspheres encapsulating
exenatide, where such pharmaceutical formulations comprise about 5%
(w/w) exenatide, about 2% (w/w) sucrose, and about 93% (w/w) of the
poly(lactide-co-glycolide) polymer.
BRIEF DESCRIPTION OF THE DRAWINGS
[0017] FIG. 1. Plasma exenatide concentrations (mean+SD) over a
period of 30 weeks in 128 patients receiving exenatide once weekly.
At week 30, the average plasma concentration was 388 pg/mL, and the
geometric mean plasma concentration was 300.2 pg/mL.
[0018] FIG. 2. Fasting Triglycerides. (2A) Fasting triglycerides
(mg/dL) decreased by an average of 42 mg/dL from baseline to week
30 in patients receiving exenatide once weekly. (2B) Fasting
triglycerides (mg/dL) decreased by an average of 22 mg/dL from
baseline to week 30 in patients receiving exenatide twice
daily.
[0019] FIG. 3. Cholesterol. (3A) Total cholesterol decreased by an
average of 11.9 mg/dL from baseline to week 30 in patients
receiving exenatide once weekly. (3B) Total cholesterol decreased
by an average of 3.8 mg/dL from baseline to week 30 in patients
receiving exenatide twice daily. (3C) LDL-cholesterol decreased by
an average of 4.9 mg/dL from baseline to week 30 in patients
receiving exenatide once weekly. (3D) LDL-cholesterol increased by
an average of 1.2 mg/dL from baseline to week 30 in patients
receiving exenatide twice daily. (3E) HDL-cholesterol decreased by
an average of 0.9 mg/dL from baseline to week 30 in patients
receiving exenatide once weekly. (3F) HDL-cholesterol decreased by
an average of 1.3 mg/dL from baseline to week 30 in patients
receiving exenatide twice daily.
[0020] FIG. 4. Weight loss. (4A) After 30 weeks, the body weight
for patients receiving exenatide once weekly decreased by an
average of 3.7 kg. (4B) After 30 weeks, the body weight for
patients receiving exenatide twice daily decreased by an average of
3.6 kg.
[0021] FIG. 5. Hemoglobin A.sub.1C (HbA.sub.1C). (A) HbA.sub.1C
decreased by an average of 1.9% from baseline to week 30 in
patients receiving exenatide once weekly. (B) HbA.sub.1C decreased
by an average of 1.5% from baseline to week 30 in patients
receiving exenatide BID.
[0022] FIG. 6. Fasting Postprandial Glucose. (A) Fasting
postprandial blood glucose decreased by an average of 42 mg/dL from
baseline to week 30 in patient receiving exenatide once weekly. (B)
Fasting postprandial blood glucose decreased by an average of 25
mg/dL from baseline to week 30 in patient receiving exenatide
BID.
DETAILED DESCRIPTION OF THE INVENTION
[0023] Provided herein are compositions, medicaments for
manufacture, and methods for reducing total cholesterol levels,
reducing LDL-cholesterol levels, reducing triglyceride levels,
treating dyslipidemia, treating atherosclerosis, and reducing the
risk of heart attacks and strokes in patients in need or desirous
thereof by administering at least one exendin, exendin agonist, or
exendin analog agonist. The methods include the chronic (i.e.,
sustained) administration of an effective amount of at least one
exendin, exendin agonist, or exendin analog agonist to a patient to
achieve the desired results as described herein.
[0024] Provided herein are methods for lowering total cholesterol
levels in patients in need thereof by identifying patients in need
of a reduction in total cholesterol levels and administering to the
patients a pharmaceutical formulation comprising a sufficient
amount of at least one exendin, exendin agonist or exendin analog
agonist to maintain a steady state average, geometric mean, or
minimum plasma concentration level of the exendin, exendin agonist,
or exendin analog agonist effective to reduce the total cholesterol
levels in the patient. The total cholesterol levels may be reduced
by at least 1 mg/dL; at least 2 mg/dL; at least 3 mg/dL; at least 4
mg/dL; at least 5 mg/dL; at least 6 mg/dL; at least 7 mg/dL; at
least 8 mg/dL; or at least 9 mg/dL. In other embodiments, the total
cholesterol levels may be reduced by at least 10 mg/dL. In still
other embodiments, total cholesterol levels may be reduced by 1
mg/dL to 25 mg/dL; from 5 mg/dL to 25 mg/dL; or from 10 mg/dL to 25
mg/dL. In one embodiment, the patient has a total blood cholesterol
level of 200 mg/dL or more. In another embodiment, the patient has
a total blood cholesterol level of 240 mg/dL or more. In one
embodiment, total blood cholesterol levels are fasting total blood
cholesterol levels that are measured after an 8-12 hour fast
without oral intake of food or liquids.
[0025] Provided herein are methods for lowering LDL-cholesterol
levels in a patients in need thereof by identifying patients in
need of a reduction in LDL-cholesterol levels and administering to
the patients a pharmaceutical formulation comprising a sufficient
amount of at least one exendin, exendin agonist or exendin analog
agonist to maintain a steady state average, geometric mean, or
minimum plasma concentration level of the exendin, exendin agonist,
or exendin analog agonist effective to reduce the LDL-cholesterol
levels in the patient. The LDL-cholesterol levels may be reduced by
at least 1 mg/dL; at least 2 mg/dL; at least 3 mg/dL; at least 4
mg/dL; at least 5 mg/dL; at least 6 mg/dL; at least 7 mg/dL; at
least 8 mg/dL; at least 9 mg/dL; or at least 10 mg/dL. In other
embodiments, LDL-cholesterol levels may be reduced by 1 mg/dL to 20
mg/dL; from 1 mg/dL to 15 mg/dL; or from 1 mg/dL to 10 mg/dL. In
one embodiment, the patient has LDL cholesterol levels of 130 mg/dL
or more. In another embodiment, the patient has LDL cholesterol
levels of 160 mg/dL or more. In yet another embodiment, the patient
has LDL cholesterol levels of 190 mg/dL or more. In one embodiment,
LDL cholesterol levels are fasting LDL cholesterol levels that are
measured after an 8-12 hour fast without oral intake of food or
liquids.
[0026] Provided herein are methods for reducing triglyceride levels
in patients in need thereof by identifying patients in need of a
reduction in triglyceride levels and administering to the patients
a pharmaceutical formulation comprising a sufficient amount of at
least one exendin, exendin agonist or exendin analog agonist to
maintain a steady state average, geometric mean, or minimum plasma
concentration level of the exendin, exendin agonist, or exendin
analog agonist effective to reduce the triglyceride levels in the
patient. In one embodiment, the triglyceride levels are fasting
triglyceride levels. Fasting triglyceride levels may be measured
after an 8-12 hour fast without oral intake of food or liquids. In
one embodiment, the triglyceride levels are reduced by at least 5
mg/dL; at least 10 mg/dL; at least 15 mg/dL; at least 20 mg/dL; at
least 25 mg/dL; at least 30 mg/dL; at least 35 mg/dL; at least 40
mg/dL; at least 45 mg/dL; or at least 50 mg/dL. In other
embodiments, the triglyceride levels are reduced from 10 mg/dL to
70 mg/dL; from 20 mg/dL to 60 mg/dL; or from 30 mg/dL to 50 mg/dL.
In one embodiment, the patient has triglyceride levels of 150 mg/dL
or higher. In another embodiment, the patient has triglyceride
levels of 200 mg/dL or higher. In yet another embodiment, the
patient has triglyceride levels of 500 mg/dL or higher.
[0027] Provided herein are methods for treating dyslipidemia in
patients in need thereof by identifying patients in need of
treatment for dyslipidemia and administering to the patients a
pharmaceutical formulation comprising a sufficient amount of at
least one exendin, exendin agonist or exendin analog agonist to
maintain a steady state average, geometric mean, or minimum plasma
concentration level of the exendin, exendin agonist, or exendin
analog agonist effective to treat dyslipidemia in the patient.
"Dyslipidemia" can include one or more of the following:
hyperlipidemia (e.g., elevated lipids), hyperglyceridemia (e.g.,
elevated glycerides), hypertriglyceridemia (e.g., elevated
triglycerides), hypercholesterolemia (e.g., elevated cholesterol),
hyperlipoproteinemia (e.g., elevated LDL-cholesterol),
hyperchylomicronemia (e.g., elevated chylomicrons), and combined
hyperlipidemia (e.g., elevated triglycerides and
LDL-cholesterol).
[0028] Provided herein are methods for treating or slowing the
progression of atherosclerosis in patients in need thereof by
identifying patients in need of treatment for atherosclerosis and
administering to the patients a pharmaceutical formulation
comprising a sufficient amount of at least one exendin, exendin
agonist or exendin analog agonist to maintain a steady state
average, geometric mean, or minimum plasma concentration level of
the exendin, exendin agonist, or exendin analog agonist effective
to treat or slow the progression of atherosclerosis in the patient.
Atherosclerosis is a disease in which plaque builds up on the
insides of arteries. The plaque is made up of fat, cholesterol,
calcium, and other substances found in blood. Over time, plaque
hardens, narrows the arteries, and reduces the flow of oxygen-rich
blood to organs and other body parts, which can lead to heart
attacks and stroke.
[0029] Provided herein are methods for treating, preventing, or
reducing the risk of heart attacks and strokes in patients in need
thereof by identifying patients in need of treatment, prevention,
or risk reduction for heart attacks or strokes and administering to
the patients a pharmaceutical formulation comprising a sufficient
amount of at least one exendin, exendin agonist or exendin analog
agonist to maintain a steady state average, geometric mean, or
minimum plasma concentration level of the exendin, exendin agonist,
or exendin analog agonist effective to treat, prevent, or reduce
the risk of heart attacks or strokes in the patient.
[0030] Provided herein are methods of treating diabetes; methods of
treating obesity; methods of reducing hemoglobin A.sub.1C; and
methods of reducing postprandial blood glucose in patients in need
thereof by administering to the patients a pharmaceutical
formulation comprising at least one exendin, exendin agonist or
exendin analog agonist in an amount sufficient to maintain a steady
state geometric mean plasma concentration of the exendin, exendin
agonist, or exendin analog agonist of about 145 pg/mL to about 700
pg/mL. In other embodiments, the steady state geometric mean plasma
concentration of the exendin, exendin agonist, or exendin analog
agonist is about 175 pg/mL to about 650 pg/mL; about 200 pg/mL to
about 600 pg/mL; about 200 pg/mL to about 400 pg/mL; or about 300
pg/mL.
[0031] In some embodiments, the exendin, exendin agonist, or
exendin analog agonist is given by chronic administration. "Chronic
administration" refers to administration of the formulations
containing the exendin, exendin agonist, or exendin analog agonist
in a continuous mode (as opposed to an acute mode) to maintain the
plasma concentration at or above the level necessary to obtain the
desired therapeutic effect for an extended period of time. In one
embodiment, the "therapeutically effective amount" is the steady
state average plasma concentration (i.e., C.sub.SS) of the exendin,
exendin agonist, or exendin analog agonist. The average plasma
concentration is the area under the plasma concentration versus
time curve during the dosing interval at steady state divided by
the dosing interval. In another embodiment, the "therapeutically
effective amount" is the steady state minimum plasma concentration
of the exendin, exendin agonist, or exendin analog agonist. In yet
another embodiment, the "therapeutically effective amount" is the
steady state geometric mean plasma concentration of the exendin,
exendin agonist, or exendin analog agonist.
[0032] The skilled artisan will recognize that the average,
geometric mean, or minimum plasma concentration needed to obtain
the desired therapeutic effect may not be reached immediately upon
administration of the exendin, exendin agonist, or exendin analog
agonist, but may take anywhere from hours to days to weeks to be
reached. Accordingly, the average, minimum, or geometric mean
plasma concentration must be measured at steady state. Once
reached, the steady state average, minimum, or geometric mean
plasma concentration is maintained for the desired period of time
to have its therapeutic effect.
[0033] "Chronic administration" also refers to administration of
the exendin, exendin agonist, or exendin analog agonist in any
manner that achieves the therapeutically effective amount.
Exemplary modes of administration include continuous infusion
(either intravenously or subcutaneously); the use of a pump or
metering system (either implanted or external, for continuous or
intermittent delivery); and extended release, slow release,
sustained release or long acting formulations. In one exemplary,
the formulation for "chronic administration" is a polymer-based
sustained release formulation. Exemplary polymer-based sustained
release formulations are described in U.S. Pat. No. 6,824,822; US
Publication No. 2006/0099271; US Publication No. 2004/0228833; US
Publication No. 2004/0208929; US Publication No. 2005/0271702; US
Publication No. 2006/0110423; WO 2004/035754; WO 2004/035762; WO
2004/036186; WO 2005/102293; WO 2005/110425; and WO 2007/081321;
the disclosures of which are incorporated by reference herein in
their entirety. Other exemplary modes of administration include
intravenous, transmucosal, intranasal, oral, intramuscular,
subcutaneous, transdermal, by inhalation, or by pulmonary
administration.
[0034] In one embodiment of the formulations and methods described
herein, the steady state minimum plasma concentration of the
exendin, exendin agonist or exendin analog agonist is at a level of
at least about 100 pg/ml, at least about 125 pg/ml, or at least
about 145 pg/mL for at least about 1 day, at least about 2 days, at
least about 3 days, at least about 1 week, at least about 2 weeks,
at least about 3 weeks, at least about 1 month, at least about 3
months, or at least about 6 months.
[0035] In other embodiments of the formulations and methods
described herein, the steady state average or minimum plasma
concentration of the exendin, exendin agonist, or exendin analog
agonist is at a level from about 100 pg/ml to about 1,000 pg/ml;
from about 100 pg/ml to about 750 pg/ml; from about 145 pg/ml to
about 700 pg/ml; from about 150 pg/ml to about 700 pg/ml; from
about 150 pg/ml to about 650 pg/ml; from about 200 pg/ml to about
650 pg/ml; from about 200 pg/mL to about 600 pg/mL; or from about
200 pg/mL to about 550 pg/mL. In one embodiment, the plasma
concentration is the steady state average plasma concentration.
[0036] In still other embodiments of the formulations and methods
described herein, the steady state average or minimum plasma
concentration of the exendin, exendin agonist, or exendin analog
agonist is at a level of least about 75 pg/ml, at least about 100
pg/ml, at least about 150 pg/ml, at least about 175 pg/ml, at least
about 200 pg/ml, at least about 225 pg/ml, or at least about 250
pg/ml. In one embodiment, the plasma concentration is the steady
state average plasma concentration.
[0037] In still other embodiments of the formulations and methods
described herein, the steady state geometric mean plasma
concentration of the exendin, exendin agonist, or exendin analog
agonist is at a level from about 145 pg/mL to about 700 pg/mL. In
other embodiments, the steady state geometric mean plasma
concentration of the exendin, exendin agonist, or exendin analog
agonist is about 175 pg/mL to about 650 pg/mL; about 200 pg/mL to
about 600 pg/mL; about 200 pg/mL to about 400 pg/mL; or about 300
pg/mL.
[0038] In still other embodiments of the formulations and methods
described herein, the steady state average or minimum plasma
concentration of the exendin, exendin agonist or exendin analog
agonist is greater than 50 pmoles/liter, greater than 60
pmoles/liter, greater than 70 pmoles/liter, greater than 80
pmoles/liter, greater than 90 pmoles/liter, greater than 100
pmoles/liter, greater than 110 pmoles/liter, greater than 120
pmoles/liter, greater than 130 pmoles/liter, greater than 140
pmoles/liter, or greater than 150 pmoles/liter. In one embodiment,
the plasma concentration is the steady state average plasma
concentration.
[0039] In further embodiments of the formulations and methods
described herein, any of the above steady state average, geometric
mean, or minimum plasma concentrations of at least one exendin,
exendin agonist, or exendin analog agonist is maintained for at
least one day, at least one week, at least one month, at least
three months, or at least one year. In other embodiments of the
formulations and methods described herein, any of the above steady
state average, geometric mean, or minimum plasma concentrations are
maintained for at least about 3 days, at least about 5 days, at
least about 1 week, at least about 2 weeks, at least about 3 weeks,
at least about one month, at least about 2 months, at least about 3
months, at least about 4 months, at least about 5 months, at least
about 6 months, at least about 7 months, at least about 8 months,
at least about 9 months, at least about 10 months, at least about
11 months or at least about one year. In one embodiment of the
formulations and methods described herein, the steady state
average, geometric mean or minimum plasma concentration is
maintained for at least about one week to about twelve weeks; from
about one week to about eight weeks; or from about one week to
about four weeks.
[0040] In any one of the embodiments of the formulations and
methods described herein, the circulating plasma exendin, exendin
agonist or exendin analog agonist concentrations may be maintained
at the steady state average, geometric mean, or minimum plasma
concentration or within about 5%, about 10%, about 20%, about 25%,
or about 50% of the average, geometric mean, or minimum plasma
concentration. In other embodiments, the circulating plasma
concentrations are maintained at the average, geometric mean, or
minimum given concentration or at about 98%, about 97%, about 96%,
about 95%, about 90%, about 80%, about 70%, or about 60% of the
average, geometric mean, or minimum concentration.
[0041] In one embodiment, the steady state average, geometric mean,
or minimum plasma concentration of the exendin, exendin agonist or
exendin analog agonist is the steady state average, geometric mean,
or minimum plasma concentration of an exendin, exendin agonist or
exendin analog agonist that results in a therapeutic effect (e.g.,
reduced total cholesterol, LDL-cholesterol, triglycerides)
substantially equivalent to that observed with a given steady state
average, geometric mean, or minimum plasma concentration of
exendin-4. Substantially equivalent may be .+-.50%, .+-.25%,
.+-.20%, .+-.10%, or .+-.5% to the steady state average, minimum,
or geometric mean plasma concentration observed with a given steady
state average, minimum, or geometric mean plasma concentration of
exendin-4.
[0042] Any formulation for sustained release of the exendin,
exendin agonist or exendin analog agonist can be used. Exemplary
formulations are described in U.S. Pat. No. 6,828,303; US
Publication Nos. 2006/0084604, 2006/0034923, 2006/0034889 and
2005/0171503; European Patent Application No. 1512395 A1; and WO
2006/041538, WO 2006/017852, WO 2005/041873, WO 2005/112633 and WO
2005/040195, the disclosures of which are incorporated by reference
herein in their entirety.
[0043] In another embodiment, the exendin, exendin agonist, or
exendin analog agonist may be administered in a polymer-based
sustained release formulation. Exemplary polymer-based sustained
release formulations include the formulations described in U.S.
Pat. No. 6,824,822; US Publication No. 2006/0099271; US Publication
No. 2004/0228833; US Publication No. 2004/0208929; US Publication
No. 2005/0271702; US Publication No. 2006/0110423; WO 2004/035754;
WO 2004/035762; WO 2004/036186; WO 2005/102293; WO 2005/110425; and
WO 2007/081321, the disclosures of which are incorporated by
reference herein in their entirety.
[0044] In some embodiments, the formulations and methods described
herein further provide that the exendin, exendin analog or exendin
analog agonist is co-administered with one or more cholesterol
and/or triglyceride lowering agents. Exemplary agents include HMG
CoA reductase inhibitors (e.g., atorvastatin, fluvastatin,
lovastatin, pravastatin, rosuvastatin, simvastatin); bile ace
sequestrants (e.g., colesevelam, cholestyramine, colestipol);
fibrates (e.g., fenofibrate, clofibrate, gemfibrozil); ezetimibe,
nicotinic acid, probucol, a lovastatin/niacin combination; an
atorvastatin/amlodipine combination; and a simvastatin/ezetimibe
combination. Co-administration can be achieved by separately
administering the exendin, exendin agonist, or exendin analog
agonist with the cholesterol and/or triglyceride lowering agent, or
be administering a single pharmaceutical formulation comprising the
exendin, exendin agonist, or exendin analog agonist and the
cholesterol and/or triglyceride lowering agent. Appropriate dosage
regimens for the cholesterol and/or triglyceride lowering agents
are generally known in the art.
[0045] The term "exendin" includes naturally occurring (or
synthetic versions of naturally occurring) exendin peptides that
are found in the salivary secretions of the Gila monster. Exendins
of particular interest include exendin-3 (SEQ ID NO:2) which is
present in the salivary secretions of Heloderma horridum, and
exendin-4 (SEQ ID NO:1) a 39 amino acid peptide which is naturally
present in the salivary secretions of Heloderma suspectum. Eng et
al, J. Biol. Chem., 265:20259-62 (1990); Eng et al, J. Biol. Chem.,
267:7402-05 (1992). Exendin-4 as it occurs in the salivary
secretions of the Gila monster is an amidated peptide. It should be
appreciated, however, that the exendin agonists and exendin analog
agonists for use in the methods described herein are not limited to
the amidated forms, but include that acid form, pharmaceutically
acceptable salt form, or any other physiologically active form of
the molecule. In one embodiment, the term exendin, as it refers to
native exendin peptides or synthetic versions of native exendin
peptides (e.g., exendin-4, exendin-3), can be used interchangeably
with the term "exendin agonist."
[0046] "Exendin agonist" and "exendin analog agonist" refer to
peptides or other compounds which elicit a biological activity of
an exendin reference peptide, preferably having a potency equal to
or better than the exendin reference peptide, or within five orders
of magnitude (plus or minus) of potency compared to the exendin
reference peptide, for example, 4, 3, 2, or 1 orders of magnitude,
when evaluated by art-known measures such as receptor binding
and/or competition studies as described, e.g., by Hargrove et al,
Regulatory Peptides, 141:113-119 (2007), the disclosure of which is
incorporated by reference herein. In one embodiment, the term
refers to a peptide or compound which elicits a biological effect
similar to that of the exendin reference peptide, for example a
compound (1) having activity in cholesterol lowering assays similar
to the exendin reference peptide, and (2) which optionally binds
specifically in a reference receptor assay or in a competitive
binding assay with labeled exendin reference peptide. Preferably,
the exendin agonists will bind in such assays with an affinity of
less than 1 .mu.M, and more preferably with an affinity of less
than 1-5 nM. Such exendin agonists may comprise a polypeptide
comprising an active fragment of a reference peptide or a small
chemical molecule. In one embodiment, the exendin agonist is a
peptide, preferably exendin-4. Exendin agonists and exendin analog
agonists do not include GLP-1 and variants, analogs and derivatives
thereof.
[0047] The term "pharmaceutically acceptable carrier" refers to a
non-toxic compound that may be administered to a patient together
with an exendin agonist or exendin analog agonist of the invention,
and which does not destroy the pharmacological activity thereof.
Pharmaceutically acceptable carriers include any and all solvents,
dispersion media, coatings, antibacterial and antifungal agents,
isotonic and absorption delaying agents and the like. The use of
such media and agents for pharmaceutically active substances is
well known in the art. Except insofar as any conventional media or
agent is incompatible with the active ingredient, its use in
therapeutic compositions is contemplated. Supplementary active
ingredients also can be incorporated into the compositions.
[0048] Exendin sequences compared to the sequence of GLP-1 are: H A
E G T F T S D V S S Y L E G Q A A K E F I A W L V K G R--NH.sub.2
(GLP-1(7-36)-NH.sub.2 (SEQ ID NO:3); H S D G T F T S D L S K Q M E
E E A V R L F I E W L K N G G P S S G A P P P S--NH.sub.2
(exendin-3-NH.sub.2 (SEQ ID NO:2)); D L S K Q M E E E A V R L F I E
W L K N G G P S S G A P P P S--NH.sub.2 (exendin-4(9-39)-NH.sub.2
(SEQ ID NO:4)); H G E G T F T S D L S K Q M E E E A V R L F I E W L
K N G G P S S G A P P P S--NH.sub.2(exendin-4-NH.sub.2 (SEQ ID
NO:1)); H S D A T F T A E Y S K L L A K L A L Q K Y L E S I L G S S
T S P R P P S S (helospectin I (SEQ ID NO:5)); H S D A T F T A E Y
S K L L A K L A L Q K Y L E S I L G S S T S P R P P S (helospectin
II (SEQ ID NO:6)); H S D A I F T E E Y S K L L A K L A L Q K Y L A
S I L G S R T S P P P--NH.sub.2 (helodermin-NH.sub.2 (SEQ ID
NO:7)): and H S D A I F T Q Q Y S K L L A K L A L Q K Y L A S I L G
S R T S P P P--NH.sub.2 (Q.sup.8,Q.sup.9 helodermin-NH.sub.2 (SEQ
ID NO:8)).
[0049] The structure activity relationship of exendin was
investigated for structures that may relate to the activity of
exendin, for its stability to metabolism, and for improvement of
its physical characteristics, especially as it pertains to peptide
stability and to amenability to alternative delivery systems, and
various exendin analog agonist peptide compounds have been
developed. Exendin analog agonists include exendin analogs with
agonist activity in which one or more naturally occurring amino
acids are inserted, eliminated or replaced with another amino
acid(s). Exemplary exendin analog agonists are peptide analogs of
exendin-4. In one aspect, the methods of reducing total cholesterol
levels, reducing LDL-cholesterol levels, reducing triglyceride
levels, and treating dyslipidemia comprise the chronic
administration of an exendin analog agonist to a patient in need
thereof.
[0050] Exendin analog agonists include peptides that are encoded by
polynucleotides that express biologically active exendin analogs
with agonist activity, as defined herein. For instance, exendin
analog agonists may be peptides containing one or more amino acid
substitutions, additions or deletions, compared with reference
exendin, for example, exendin-4. In one embodiment, the number of
substitutions, deletions, or additions is 15 amino acids or less,
10 amino acids or less, 5 amino acids or less, 3 amino acids or
less, 2 amino acids or less, or 1 amino acid or less. In one aspect
of the invention, the substitutions include one or more
conservative substitutions. A "conservative" substitution denotes
the replacement of an amino acid residue by another, biologically
active, similar residue. Examples of conservative substitutions
include the substitution of one hydrophobic residue, such as
isoleucine, valine, leucine, or methionine for another, or the
substitution of one polar residue for another, such as the
substitution of arginine for lysine, glutamic for aspartic acids,
or glutamine for asparagine, and the like. The following table
lists illustrative, but non-limiting, conservative amino acid
substitutions.
TABLE-US-00001 TABLE ORIGINAL RESIDUE EXEMPLARY SUBSTITUTIONS ALA
SER, THR ARG LYS ASN HIS, SER ASP GLU, ASN CYS SER GLN ASN, HIS GLU
ASP GLY ALA, SER HIS ASN, GLN ILE LEU, VAL, THR LEU ILE, VAL LYS
ARG, GLN, GLU, THR MET LEU, ILE, VAL PHE LEU, TYR SER THR, ALA, ASN
THR SER, ALA TRP ARG, SER TYR PHE VAL ILE, LEU, ALA PRO ALA
[0051] It is further understood that exendin analog agonists
include the peptides described herein which have been chemically
derivatized or altered, for example, peptides with non-natural
amino acid residues (e.g., taurine, .beta.-amino acid residues,
.gamma.-amino acid residues, and D-amino acid residues), C-terminal
functional group modifications, such as amides, esters, and
C-terminal ketone modifications and N-terminal functional group
modifications, such as acylated amines, Schiff bases, or
cyclization, as found, for example, in the amino acid pyroglutamic
acid.
[0052] Such derivatized peptides include exendins, exendin agonists
and exendin analog agonists conjugated (with or without a linking
group) to one or more polymer molecules, such as polyethylene
glycol ("PEG"), albumin, fatty acid chains of various lengths
(e.g., stearyl, palmitoyl, octanoyl, etc.), or by the addition of
polyamino acids, such as poly-his, poly-arg, poly-lys, and
poly-ala. Modifications to the exendins, exendin agonists and
exendin analog agonists can also include small molecule
substituents, such as short alkyls and constrained alkyls (e.g.,
branched, cyclic, fused, adamantyl), and aromatic groups. The
polymer molecules will typically have a molecular weight ranging
from about 500 to about 20,000 Daltons. Albumin has a molecular
weight of about 67,000 Daltons. Such peptides are described, for
example, in U.S. Pat. No. 6,924,264, and 6,593,295, the disclosure
of which are incorporated by reference herein in their entirety.
Exemplary modified peptides include
(His).sub.x-[Exendin-4(1-39)]-(Lys).sub.y where x is 0, 1 or 2; and
y is an integer from 2-8; and [Exendin-4(1-39)]-(linking
group)-albumin, where the linking group comprises (i) one or more
Lys amino acids adjacent the C-terminal end of the exendin 4, (ii)
a maleimide group, (iii) a --(OCH.sub.2CH.sub.2).sub.z-- group
where z is 1, 2, 3, or 4; or (iv) a combination thereof.
[0053] Such polymer-conjugations and small molecule substituent
modifications may occur singularly at the N- or C-terminus or at
the side chains of amino acid residues within the sequence of the
polypeptides. Alternatively, there may be multiple sites of
derivatization along the hybrid polypeptide. Substitution of one or
more amino acids with lysine, aspartic acid, glutamic acid, or
cysteine may provide additional sites for derivatization. See,
e.g., U.S. Pat. Nos. 5,824,784 and 5,824,778. In one embodiment,
the polypeptides may be conjugated to one, two, or three polymer
molecules.
[0054] The polymer molecules may be linked to an amino, carboxyl,
or thiol group, and may be linked by N or C terminus, or at the
side chains of lysine, aspartic acid, glutamic acid, or cysteine.
Alternatively, the polymer molecules may be linked with diamine and
dicarboxylic groups. In one embodiment, the polypeptides are
conjugated to one, two, or three PEG molecules through an epsilon
amino group on a lysine amino acid.
[0055] Also included in the invention are exendin analog sequences
having 80% sequence identity, 90% sequence identity, 95% sequence
identity, or 99% sequence identity to a reference exendin peptide,
for example, (1) SEQ ID NO:1; (2) SEQ ID NO:2; or (2) truncated
sequences of SEQ ID NO:1 or SEQ ID NO:2, wherein the truncated
sequences contain at least 10 amino acids, at least 20 amino acids,
at least 25 amino acids, at least 30 amino acids, at least 35 amino
acids, at least 38 amino acids or N-1 amino acids where N equals
the number of amino acids in the full length or reference peptide
or protein. As used herein, sequence identity refers to a
comparison made between two molecules using standard algorithms
well known in the art. In one embodiment, the algorithm for
calculating sequence identity for the invention is the
Smith-Waterman algorithm, where an exendin, for example SEQ ID NO:1
(i.e., exendin-4), is used as the reference sequence to define the
percentage identity of a comparison peptide over its length. The
choice of parameter values for matches, mismatches, and insertions
or deletions is discretionary, although some parameter values have
been found to yield more biologically realistic results than
others. In one embodiment, the set of parameter values for the
Smith-Waterman algorithm is set forth in the "maximum similarity
segments" approach, which uses values of 1 for a matched residue
and -1/3 for a mismatched residue (a residue being either a single
nucleotide or single amino acid). Waterman, Bull. Math. Biol. 46;
473 (1984). Insertions and deletions (indels), x, are weighted as
x.sub.k=1+1/3k, where k is the number of residues in a given
insertion or deletion. Id.
[0056] Other exendins, exendin agonists, and exendin analog
agonists useful in the invention include those described in WO
98/05351; WO 99/07404; WO 99/25727; WO 99/25728; WO 99/40788; WO
00/41546; WO 00/41548; WO 00/73331; WO 01/51078; WO 03/099314; U.S.
Pat. No. 6,956,026; U.S. Pat. No. 6,506,724; U.S. Pat. No.
6,703,359; U.S. Pat. No. 6,858,576; U.S. Pat. No. 6,872,700; U.S.
Pat. No. 6,902,744; U.S. Pat. No. 7,157,555; U.S. Pat. No.
7,223,725; U.S. Pat. No. 7,220,721; US Publication No.
2003/0036504; and US Publication No. 2006/0094652, the disclosures
of which are incorporated by reference herein in their
entirety.
[0057] Certain exemplary exendins, exendin agonists, and exendin
analog agonists include: exendin-4 (1-30) (SEQ ID NO:9: 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:10: His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Met Glu Glu Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly
Gly-NH.sub.2); exendin-4 (1-28) amide (SEQ ID NO:11: 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.sub.2); .sup.14Leu,.sup.25Phe
exendin-4 amide (SEQ ID NO:12: His Gly Glu Gly Thr Phe Thr Ser Asp
Leu Ser Lys Gln Leu Glu Glu Glu Ala Val Arg Leu Phe Ile Glu Phe Leu
Lys Asn Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser-NH.sub.2);
.sup.14Leu,.sup.25Phe exendin-4 (1-28) amide (SEQ ID NO:13: His Gly
Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu Glu Ala Val
Arg Leu Phe Ile Glu Phe Leu Lys Asn-NH.sub.2); and
.sup.14Leu,.sup.22Ala,.sup.25Phe exendin-4 (1-28) amide (SEQ ID
NO:14: His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu
Glu Glu Ala Val Arg Leu Ala Ile Glu Phe Leu Lys Asn-NH.sub.2).
[0058] Exendin analog agonists for use in the formulations and
methods described herein also include those described in U.S. Pat.
No. 7,223,725, such as compounds of formula (I) (SEQ ID NO:15) or a
pharmaceutically acceptable salt thereof: Xaa.sub.1 Xaa.sub.2
Xaa.sub.3 Gly Xaa.sub.5 Xaa.sub.6 Xaa.sub.7 Xaa.sub.8 Xaa.sub.9
Xaa.sub.10 Xaa.sub.11 Xaa.sub.12 Xaa.sub.13 Xaa.sub.14 Xaa.sub.15
Xaa.sub.16 Xaa.sub.17 Ala Xaa.sub.19 Xaa.sub.20 Xaa.sub.21
Xaa.sub.22 Xaa.sub.23 Xaa.sub.24 Xaa.sub.25 Xaa.sub.26 Xaa.sub.27
Xaa.sub.28-Z.sub.1; wherein Xaa.sub.1 is His, Arg or Tyr; Xaa.sub.2
is Ser, Gly, Ala or Thr; Xaa.sub.3 is Ala, Asp or Glu; Xaa.sub.5 is
Ala or Thr; Xaa.sub.6 is Ala, Phe, Tyr or naphthylalanine;
Xaa.sub.7 is Thr or Ser; Xaa.sub.8 is Ala, Ser or Thr; Xaa.sub.9 is
Asp or Glu; Xaa.sub.10 is Ala, Leu, Ile, Val, pentylglycine or Met;
Xaa.sub.11 is Ala or Ser; Xaa.sub.12 is Ala or Lys; Xaa.sub.13 is
Ala or Gln; Xaa.sub.14 is Ala, Leu, Ile, pentylglycine, Val or Met;
Xaa.sub.15 is Ala or Glu; Xaa.sub.16 is Ala or Glu; Xaa.sub.17 is
Ala or Glu; Xaa.sub.19 is Ala or Val; Xaa.sub.20 is Ala or Arg;
Xaa.sub.21 is Ala or Leu; Xaa.sub.22 is Ala, Phe, Tyr or
naphthylalanine; Xaa.sub.23 is Ile, Val, Leu, pentylglycine,
tert-butylglycine or Met; Xaa.sub.24 is Ala, Glu or Asp; Xaa.sub.25
is Ala, Trp, Phe, Tyr or naphthylalanine; Xaa.sub.26 is Ala or Leu;
Xaa.sub.27 is Ala or Lys; Xaa.sub.28 is Ala or Asn; Z.sub.1 is
--OH, --NH.sub.2, Gly-Z.sub.2, Gly Gly-Z.sub.2, Gly Gly
Xaa.sub.31-Z.sub.2, Gly Gly Xaa.sub.31 Ser-Z.sub.2, Gly Gly
Xaa.sub.31 Ser Ser-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser Gly-Z.sub.2,
Gly Gly Xaa.sub.31 Ser Ser Gly Ala-Z.sub.2, Gly Gly Xaa.sub.31 Ser
Ser Gly Ala Xaa.sub.36-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser Gly Ala
Xaa.sub.36 Xaa.sub.37-Z.sub.2 or Gly Gly Xaa.sub.31 Ser Ser Gly Ala
Xaa.sub.36 Xaa.sub.37 Xaa.sub.38-Z.sub.2; Xaa.sub.31, Xaa.sub.36,
Xaa.sub.37 and Xaa.sub.38 are independently Pro, homoproline, 3Hyp,
4Hyp, thioproline, N-alkylglycine, N-alkylpentylglycine or
N-alkylalanine; and Z.sub.2 is --OH or --NH.sub.2; provided that no
more than three of Xaa.sub.3, Xaa.sub.5, Xaa.sub.6, Xaa.sub.8,
Xaa.sub.10, Xaa.sub.11, Xaa.sub.12, Xaa.sub.13, Xaa.sub.14,
Xaa.sub.15, Xaa.sub.16, Xaa.sub.17, Xaa.sub.19, Xaa.sub.20,
Xaa.sub.21, Xaa.sub.24, Xaa.sub.25, Xaa.sub.26, Xaa.sub.27 and
Xaa.sub.28 are Ala.
[0059] Exemplary N-alkyl groups for N-alkylglycine,
N-alkylpentylglycine and N-alkylalanine include lower alkyl groups,
for example, of 1 to about 6 carbon atoms, or of 1 to 4 carbon
atoms. Exemplary exendin analogs include those wherein Xaa.sub.1 is
His or Tyr. More preferably Xaa.sub.1 is His. Provided are those
compounds wherein Xaa.sub.2 is Gly. Provided are those compounds
wherein Xaa.sub.14 is Leu, pentylglycine or Met. Exemplary
compounds are those wherein Xaa.sub.25 is Trp or Phe. Exemplary
compounds are those where Xaa.sub.6 is Phe or naphthylalanine;
Xaa.sub.22 is Phe or naphthylalanine and Xaa.sub.23 is Ile or Val.
Provided are compounds wherein Xaa.sub.31, Xaa.sub.36, Xaa.sub.37
and Xaa.sub.38 are independently selected from Pro, homoproline,
thioproline and N-alkylalanine. Preferably Z.sub.1 is --NH.sub.2.
Preferably Z.sub.2 is --NH.sub.2.
[0060] According to one embodiment, provided are compounds of
formula (I) wherein Xaa.sub.1 is His or Tyr, more preferably His;
Xaa.sub.2 is Gly; Xaa.sub.6 is Phe or naphthylalanine; Xaa.sub.14
is Leu, pentylglycine or Met; Xaa.sub.22 is Phe or naphthylalanine;
Xaa.sub.23 is Ile or Val; Xaa.sub.31, Xaa.sub.36, Xaa.sub.37 and
Xaa.sub.38 are independently selected from Pro, homoproline,
thioproline or N-alkylalanine. More preferably Z.sub.1 is
--NH.sub.2.
[0061] According to one embodiment, exemplary compounds include
those of formula (I) wherein: Xaa.sub.1 is His or Arg; Xaa.sub.2 is
Gly or Ala; Xaa.sub.3 is Asp or Glu; Xaa.sub.5 is Ala or Thr;
Xaa.sub.6 is Ala, Phe or naphthylalanine; Xaa.sub.7 is Thr or Ser;
Xaa.sub.8 is Ala, Ser or Thr; Xaa.sub.9 is Asp or Glu; Xaa.sub.10
is Ala, Leu or pentylglycine; Xaa.sub.11 is Ala or Ser; Xaa.sub.12
is Ala or Lys; Xaa.sub.13 is Ala or Gln; Xaa.sub.14 is Ala, Leu or
pentylglycine; Xaa.sub.15 is Ala or Glu; Xaa.sub.16 is Ala or Glu;
Xaa.sub.17 is Ala or Glu; Xaa.sub.19 is Ala or Val; Xaa.sub.20 is
Ala or Arg; Xaa.sub.21 is Ala or Leu; Xaa.sub.22 is Phe or
naphthylalanine; Xaa.sub.23 is Ile, Val or tert-butylglycine;
Xaa.sub.24 is Ala, Glu or Asp; Xaa.sub.25 is Ala, Trp or Phe;
Xaa.sub.26 is Ala or Leu; Xaa.sub.27 is Ala or Lys; Xaa.sub.28 is
Ala or Asn; Z.sub.1 is --OH, --NH.sub.2, Gly-Z.sub.2, Gly
Gly-Z.sub.2, Gly Gly Xaa.sub.31-Z.sub.2, Gly Gly Xaa.sub.31
Ser-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser-Z.sub.2, Gly Gly Xaa.sub.31
Ser Ser Gly-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser Gly Ala-Z.sub.2,
Gly Gly Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36-Z.sub.2, Gly Gly
Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37-Z.sub.2, Gly Gly
Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37
Xaa.sub.38-Z.sub.2; Xaa.sub.31, Xaa.sub.36, Xaa.sub.37 and
Xaa.sub.38 being independently Pro homoproline, thioproline or
N-methylalanine; and Z.sub.2 being --OH or --NH.sub.2; provided
that no more than three of Xaa.sub.3, Xaa.sub.5, Xaa.sub.6,
Xaa.sub.8, Xaa.sub.10, Xaa.sub.11, Xaa.sub.12, Xaa.sub.13,
Xaa.sub.14, Xaa.sub.15, Xaa.sub.16, Xaa.sub.17, Xaa.sub.19,
Xaa.sub.20, Xaa.sub.21, Xaa.sub.24, Xaa.sub.25, Xaa.sub.26,
Xaa.sub.27 and Xaa.sub.28 are Ala. Other exemplary compounds
include those set forth in WO 99/25727 identified therein as
compounds 2-23.
[0062] According to another embodiment, provided are compounds
where Xaa.sub.14 is Leu, Ile, Val or pentylglycine, more preferably
Leu or pentylglycine, and Xaa.sub.25 is Phe, Tyr or
naphthylalanine, more preferably Phe or naphthylalanine. These
compounds will be less susceptive to oxidative degradation, both in
vitro and in vivo, as well as during synthesis of the compound.
[0063] Exendin analog agonists for use in the formulations and
methods described herein also include those described in U.S. Pat.
No. 7,220,721, such as compounds of formula (II) (SEQ ID NO:16) or
a pharmaceutically acceptable salt thereof: Xaa.sub.1 Xaa.sub.2
Xaa.sub.3 Xaa.sub.4 Xaa.sub.5 Xaa.sub.6 Xaa.sub.7 Xaa.sub.8
Xaa.sub.9 Xaa.sub.10 Xaa.sub.11 Xaa.sub.12 Xaa.sub.13 Xaa.sub.14
Xaa.sub.15 Xaa.sub.16 Xaa.sub.17 Ala Xaa.sub.19 Xaa.sub.20
Xaa.sub.21 Xaa.sub.22 Xaa.sub.23 Xaa.sub.24 Xaa.sub.25 Xaa.sub.26
Xaa.sub.27 Xaa.sub.28-Z.sub.1; wherein: Xaa.sub.1 is His, Arg, Tyr,
Ala, Norval, Val or Norleu; Xaa.sub.2 is Ser, Gly, Ala or Thr;
Xaa.sub.3 is Ala, Asp or Glu; Xaa.sub.4 is Ala, Norval, Val, Norleu
or Gly; Xaa.sub.5 is Ala or Thr; Xaa.sub.6 is Ala, Phe, Tyr or
naphthylalanine; Xaa.sub.7 is Thr or Ser; Xaa.sub.8 is Ala, Ser or
Thr; Xaa.sub.9 is Ala, Norval, Val, Norleu, Asp or Glu; Xaa.sub.10
is Ala, Leu, Ile, Val, pentylglycine or Met; Xaa.sub.11 is Ala or
Ser; Xaa.sub.12 is Ala or Lys; Xaa.sub.13 is Ala or Gln; Xaa.sub.14
is Ala, Leu, Ile, pentylglycine, Val or Met; Xaa.sub.15 is Ala or
Glu; Xaa.sub.16 is Ala or Glu; Xaa.sub.17 is Ala or Glu; Xaa.sub.19
is Ala or Val; Xaa.sub.20 is Ala or Arg; Xaa.sub.21 is Ala or Leu;
Xaa.sub.22 is Phe, Tyr or naphthyl-alanine; Xaa.sub.23 is Ile, Val,
Leu, pentylglycine, tert-butylglycine or Met; Xaa.sub.24 is Ala,
Glu or Asp; Xaa.sub.25 is Ala, Trp, Phe, Tyr or naphthylalanine;
Xaa.sub.26 is Ala or Leu; Xaa.sub.27 is Ala or Lys; Xaa.sub.28 is
Ala or Asn; Z.sub.1 is --OH, --NH.sub.2, Gly-Z.sub.2, Gly
Gly-Z.sub.2, Gly Gly Xaa.sub.31-Z.sub.2, Gly Gly Xaa.sub.31
Ser-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser-Z.sub.2, Gly Gly Xaa.sub.31
Ser Ser Gly-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser Gly Ala-Z.sub.2,
Gly Gly Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36-Z.sub.2, Gly Gly
Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37-Z.sub.2, Gly Gly
Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37 Xaa.sub.38-Z.sub.2
or Gly Gly Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37
Xaa.sub.38 Xaa.sub.39-Z.sub.2; Xaa.sub.31, Xaa.sub.36, Xaa.sub.37
and Xaa.sub.38 are independently Pro, homoproline, 3Hyp, 4Hyp,
thioproline, N-alkylglycine, N-alkylpentylglycine or
N-alkylalanine; Xaa.sub.39 is Ser or Tyr; and Z.sub.2 is --OH or
--NH.sub.2; provided that no more than three of Xaa.sub.3,
Xaa.sub.4, Xaa.sub.5, Xaa.sub.6, Xaa.sub.8, Xaa.sub.9, Xaa.sub.10,
Xaa.sub.11, Xaa.sub.12, Xaa.sub.13, Xaa.sub.14, Xaa.sub.15,
Xaa.sub.16, Xaa.sub.17, Xaa.sub.19, Xaa.sub.20, Xaa.sub.21,
Xaa.sub.24, Xaa.sub.25, Xaa.sub.26, Xaa.sub.27 and Xaa.sub.28 are
Ala; and provided also that, if Xaa.sub.1 is His, Arg or Tyr, then
at least one of Xaa.sub.3, Xaa.sub.4 and Xaa.sub.9 is Ala.
[0064] Exemplary N-alkyl groups for N-alkylglycine,
N-alkylpentylglycine and N-alkylalanine include lower alkyl groups,
for example, of 1 to about 6 carbon atoms, or of 1 to 4 carbon
atoms. In one embodiment, such exendin analogs include those
wherein Xaa.sub.1 is His, Ala or Norval. More preferably Xaa.sub.1
is His or Ala. Most preferably Xaa.sub.1 is His. Provided are those
compounds of formula (II) wherein Xaa.sub.2 is Gly. Provided are
those compounds of formula (II) wherein Xaa.sub.3 is Ala. Provided
are those compounds of formula (II) wherein Xaa.sub.4 is Ala.
Provided are those compounds of formula (II) wherein Xaa.sub.9 is
Ala. Provided are those compounds of formula (II) wherein
Xaa.sub.14 is Leu, pentylglycine or Met. Exemplary compounds of
formula (II) are those wherein Xaa.sub.25 is Trp or Phe. Exemplary
compounds of formula (II) are those where Xaa.sub.6 is Ala, Phe or
naphthylalanine; Xaa.sub.22 is Phe or naphthylalanine; and
Xaa.sub.23 is Ile or Val. Provided are compounds of formula (II)
wherein Xaa.sub.31, Xaa.sub.36, Xaa.sub.37 and Xaa.sub.38 are
independently selected from Pro, homoproline, thioproline and
N-alkylalanine. Preferably Z.sub.1 is --NH.sub.2. Preferably
Z.sub.2 is --NH.sub.2.
[0065] According to one embodiment, provided are compounds of
formula (II) wherein Xaa.sub.1 is Ala, His or Tyr, more preferably
Ala or His; Xaa.sub.2 is Ala or Gly; Xaa.sub.6 is Phe or
naphthylalanine; Xaa.sub.14 is Ala, Leu, pentylglycine or Met;
Xaa.sub.22 is Phe or naphthylalanine; Xaa.sub.23 is Ile or Val;
Xaa.sub.31, Xaa.sub.36, Xaa.sub.37 and Xaa.sub.38 are independently
selected from Pro, homoproline, thioproline or N-alkylalanine; and
Xaa.sub.39 is Ser or Tyr, more preferably Ser. More preferably
Z.sub.1 is --NH.sub.2.
[0066] According to another embodiment, exemplary compounds include
those of formula (II) wherein: Xaa.sub.1 is His or Ala; Xaa.sub.2
is Gly or Ala; Xaa.sub.3 is Ala, Asp or Glu; Xaa.sub.4 is Ala or
Gly; Xaa.sub.5 is Ala or Thr; Xaa.sub.6 is Phe or naphthylalanine;
Xaa.sub.7 is Thr or Ser; Xaa.sub.8 is Ala, Ser or Thr; Xaa.sub.9 is
Ala, Asp or Glu; Xaa.sub.10 is Ala, Leu or pentylglycine;
Xaa.sub.11 is Ala or Ser; Xaa.sub.12 is Ala or Lys; Xaa.sub.13 is
Ala or Gln; Xaa.sub.14 is Ala, Leu, Met or pentylglycine;
Xaa.sub.15 is Ala or Glu; Xaa.sub.16 is Ala or Glu; Xaa.sub.17 is
Ala or Glu; Xaa.sub.19 is Ala or Val; Xaa.sub.20 is Ala or Arg;
Xaa.sub.21 is Ala or Leu; Xaa.sub.22 is Phe or naphthylalanine;
Xaa.sub.23 is Ile, Val or tert-butylglycine; Xaa.sub.24 is Ala, Glu
or Asp; Xaa.sub.25 is Ala, Trp or Phe; Xaa.sub.26 is Ala or Leu;
Xaa.sub.27 is Ala or Lys; Xaa.sub.28 is Ala or Asn; Z.sub.1 is
--OH, --NH.sub.2, Gly-Z.sub.2, Gly Gly-Z.sub.2, Gly Gly
Xaa.sub.31-Z.sub.2, Gly Gly Xaa.sub.31 Ser-Z.sub.2, Gly Gly
Xaa.sub.31 Ser Ser-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser Gly-Z.sub.2,
Gly Gly Xaa.sub.31 Ser Ser Gly Ala-Z.sub.2, Gly Gly Xaa.sub.31 Ser
Ser Gly Ala Xaa.sub.36-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser Gly Ala
Xaa.sub.36 Xaa.sub.37-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser Gly Ala
Xaa.sub.36 Xaa.sub.37 Xaa.sub.38-Z.sub.2 or Gly Gly Xaa.sub.31 Ser
Ser Gly Ala Xaa.sub.36 Xaa.sub.37 Xaa.sub.38 Xaa.sub.39-Z.sub.2;
Xaa.sub.31, Xaa.sub.36, Xaa.sub.37 and Xaa.sub.38 being
independently Pro homoproline, thioproline or N-methylalanine; and
Z.sub.2 being --OH or --NH.sub.2; provided that no more than three
of Xaa.sub.3, Xaa.sub.5, Xaa.sub.6, Xaa.sub.8, Xaa.sub.10,
Xaa.sub.11, Xaa.sub.12, Xaa.sub.13, Xaa.sub.14, Xaa.sub.15,
Xaa.sub.16, Xaa.sub.17, Xaa.sub.19, Xaa.sub.20, Xaa.sub.21,
Xaa.sub.24, Xaa.sub.25, Xaa.sub.26, Xaa.sub.27 and Xaa.sub.28 are
Ala; and provided also that, if Xaa.sub.1 is His, Arg or Tyr, then
at least one of Xaa.sub.3, Xaa.sub.4 and Xaa.sub.9 is Ala.
[0067] According to still another embodiment, provided are
compounds of formula (II) where Xaa.sub.14 is Ala, Leu, Ile, Val or
pentylglycine, more preferably Leu or pentylglycine, and Xaa.sub.25
is Ala, Phe, Tyr or naphthylalanine, more preferably Phe or
naphthylalanine. These compounds will be less susceptible to
oxidative degradation, both in vitro and in vivo, as well as during
synthesis of the compound.
[0068] Exemplary compounds of formula (II) include those described
in WO 99/25728 as having the amino acid sequence of SEQ ID NOs:
5-93 therein which are hereby incorporated by reference.
[0069] In other embodiments of the formulations and methods
described herein, the exendin analog includes narrower genera of
U.S. Pat. No. 7,223,725, having formula (III) (SEQ ID NO:17) or a
pharmaceutically acceptable salt thereof: Xaa.sub.1 Xaa.sub.2
Xaa.sub.3 Gly Xaa.sub.5 Xaa.sub.6 Xaa.sub.7 Xaa.sub.8 Xaa.sub.9
Xaa.sub.10 Xaa.sub.11 Xaa.sub.12 Xaa.sub.13 Xaa.sub.14 Xaa.sub.15
Xaa.sub.16 Xaa.sub.17 Ala Xaa.sub.19 Xaa.sub.20 Xaa.sub.21
Xaa.sub.22 Xaa.sub.23 Xaa.sub.24 Xaa.sub.25 Xaa.sub.26 Xaa.sub.27
Xaa.sub.28-Z.sub.1; wherein: Xaa.sub.1 is His or Arg; Xaa.sub.2 is
Gly or Ala; Xaa.sub.3 is Ala, Asp or Glu; Xaa.sub.5 is Ala or Thr;
Xaa.sub.6 is Ala, Phe or naphthylalanine; Xaa.sub.7 is Thr or Ser;
Xaa.sub.8 is Ala, Ser or Thr; Xaa.sub.9 is Asp or Glu; Xaa.sub.10
is Ala, Leu or pentylglycine; Xaa.sub.11 is Ala or Ser; Xaa.sub.12
is Ala or Lys; Xaa.sub.13 is Ala or Gln; Xaa.sub.14 is Ala, Leu or
pentylglycine; Xaa.sub.15 is Ala or Glu; Xaa.sub.16 is Ala or Glu;
Xaa.sub.17 is Ala or Glu; Xaa.sub.19 is Ala or Val; Xaa.sub.20 is
Ala or Arg; Xaa.sub.21 is Ala or Leu; Xaa.sub.22 is Phe or
naphthylalanine; Xaa.sub.23 is Ile, Val or tert-butylglycine;
Xaa.sub.24 is Ala, Glu or Asp; Xaa.sub.25 is Ala, Trp, or Phe;
Xaa.sub.26 is Ala or Leu; Xaa.sub.27 is Ala or Lys; Xaa.sub.28 is
Ala or Asn; Z.sub.1 is --OH, --NH.sub.2, Gly-Z.sub.2, Gly
Gly-Z.sub.2, Gly Gly Xaa.sub.31-Z.sub.2, Gly Gly Xaa.sub.31
Ser-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser-Z.sub.2, Gly Gly Xaa.sub.31
Ser Ser Gly-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser Gly Ala-Z.sub.2,
Gly Gly Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36-Z.sub.2, Gly Gly
Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37-Z.sub.2 or Gly Gly
Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37
Xaa.sub.38-Z.sub.2; Xaa.sub.31, Xaa.sub.36, Xaa.sub.37 and
Xaa.sub.38 are independently selected from the group consisting of
Pro, homoproline, thioproline and N-methylalanine; and Z.sub.2 is
--OH or --NH.sub.2; provided that no more than three of Xaa.sub.3,
Xaa.sub.5, Xaa.sub.6, Xaa.sub.8, Xaa.sub.10, Xaa.sub.11,
Xaa.sub.12, Xaa.sub.13, Xaa.sub.14, Xaa.sub.15, Xaa.sub.16,
Xaa.sub.17, Xaa.sub.19, Xaa.sub.20, Xaa.sub.21, Xaa.sub.24,
Xaa.sub.25, Xaa.sub.26, Xaa.sub.27 and Xaa.sub.28 are Ala; and
pharmaceutically acceptable salts thereof.
[0070] In other embodiments of the formulations and methods
described herein, the exendin analog includes narrower genera of
compounds described in U.S. Pat. No. 7,220,721, having formula (IV)
(SEQ ID NO:18) or a pharmaceutically acceptable salt thereof:
Xaa.sub.1 Xaa.sub.2 Xaa.sub.3 Xaa.sub.4 Xaa.sub.5 Xaa.sub.6
Xaa.sub.7 Xaa.sub.8 Xaa.sub.9 Xaa.sub.10 Xaa.sub.11 Xaa.sub.12
Xaa.sub.13 Xaa.sub.14 Xaa.sub.15 Xaa.sub.16 Xaa.sub.17 Ala
Xaa.sub.19 Xaa.sub.20 Xaa.sub.21 Xaa.sub.22 Xaa.sub.23 Xaa.sub.24
Xaa.sub.25 Xaa.sub.26 Xaa.sub.27 Xaa.sub.28-Z.sub.1; wherein:
Xaa.sub.1 is His or Ala; Xaa.sub.2 is Gly or Ala; Xaa.sub.3 is Ala,
Asp or Glu; Xaa.sub.4 is Ala or Gly; Xaa.sub.5 is Ala or Thr;
Xaa.sub.6 is Ala, Phe or naphthylalanine; Xaa.sub.7 is Thr or Ser;
Xaa.sub.8 is Ala, Ser or Thr; Xaa.sub.9 is Ala, Asp or Glu;
Xaa.sub.10 is Ala, Leu or pentylglycine; Xaa.sub.11 is Ala or Ser;
Xaa.sub.12 is Ala or Lys; Xaa.sub.13 is Ala or Gln; Xaa.sub.14 is
Ala, Leu, Met or pentylglycine; Xaa.sub.15 is Ala or Glu;
Xaa.sub.16 is Ala or Glu; Xaa.sub.17 is Ala or Glu; Xaa.sub.19 is
Ala or Val; Xaa.sub.20 is Ala or Arg; Xaa.sub.21 is Ala or Leu;
Xaa.sub.22 is Phe or naphthyl-alanine; Xaa.sub.23 is Ile, Val or
tert-butylglycine; Xaa.sub.24 is Ala, Glu or Asp; Xaa.sub.25 is
Ala, Trp or Phe; Xaa.sub.26 is Ala or Leu; Xaa.sub.27 is Ala or
Lys; Xaa.sub.28 is Ala or Asn; Z.sub.1 is --OH, --NH.sub.2,
Gly-Z.sub.2, Gly Gly-Z.sub.2, Gly Gly Xaa.sub.31-Z.sub.2, Gly Gly
Xaa.sub.31 Ser-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser-Z.sub.2, Gly Gly
Xaa.sub.31 Ser Ser Gly-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser Gly
Ala-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36-Z.sub.2,
Gly Gly Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37-Z.sub.2,
Gly Gly Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37
Xaa.sub.38-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36
Xaa.sub.37 Xaa.sub.38 Ser-Z.sub.2; Xaa.sub.31, Xaa.sub.36,
Xaa.sub.37 and Xaa.sub.38 are independently Pro, homoproline,
thioproline, or N-methylalanine; and Z.sub.2 is --OH or --NH.sub.2;
provided that no more than three of Xaa.sub.3, Xaa.sub.5,
Xaa.sub.6, Xaa.sub.8, Xaa.sub.10, Xaa.sub.11, Xaa.sub.12,
Xaa.sub.13, Xaa.sub.14, Xaa.sub.15, Xaa.sub.16, Xaa.sub.17,
Xaa.sub.19, Xaa.sub.20, Xaa.sub.21, Xaa.sub.24, Xaa.sub.25,
Xaa.sub.26, Xaa.sub.27, and Xaa.sub.28 are Ala; and provided that,
if Xaa.sub.1 is His, Arg or Tyr, then at least one of Xaa.sub.3,
Xaa.sub.4 and Xaa.sub.9 is Ala; and pharmaceutically acceptable
salts thereof.
[0071] Exemplary compounds of formula (IV) include those wherein
Xaa.sub.1 is His or Ala. Preferably, Xaa.sub.1 is His. Exemplary
compounds of formula (IV) include those wherein Xaa.sub.2 is Gly.
Exemplary compounds of formula (IV) include those wherein Xaa.sub.4
is Ala. Exemplary compounds of formula (IV) include those wherein
Xaa.sub.9 is Ala. Exemplary compounds of formula (IV) include those
wherein Xaa.sub.14 is Leu, pentylglycine or Met. Exemplary
compounds of formula (IV) include those wherein Xaa.sub.25 is Trp
or Phe. Exemplary compounds of formula (IV) include those wherein
Xaa.sub.6 is Ala, Phe or naphthylalanine; Xaa.sub.22 is Phe or
naphthylalanine; and Xaa.sub.23 is Ile or Val. Exemplary compounds
of formula (IV) include those wherein Z.sub.1 is --NH.sub.2.
Exemplary compounds of formula (IV) include those wherein
Xaa.sub.31, Xaa.sub.36, Xaa.sub.37 and Xaa.sub.38 are independently
selected from the group consisting of Pro, homoproline, thioproline
and N-alkylalanine. Exemplary compounds of formula (IV) include
those wherein Z.sub.2 is --NH.sub.2. Exemplary compounds of formula
(IV) include those wherein Z.sub.1 is --NH.sub.2.
[0072] Also provided are compounds for use in the formulations and
methods described herein are those described in U.S. Pat. No.
7,223,725, including compounds of formula (V) (SEQ ID NO:19) or a
pharmaceutically acceptable salt thereof: Xaa.sub.1 Xaa.sub.2
Xaa.sub.3 Gly Xaa.sub.5 Xaa.sub.6 Xaa.sub.7 Xaa.sub.8 Xaa.sub.9
Xaa.sub.10 Xaa.sub.11 Xaa.sub.12 Xaa.sub.13 Xaa.sub.14 Xaa.sub.15
Xaa.sub.16 Xaa.sub.17 Ala Xaa.sub.19 Xaa.sub.20 Xaa.sub.21
Xaa.sub.22 Xaa.sub.23 Xaa.sub.24 Xaa.sub.25 Xaa.sub.26 Xaa.sub.27
Xaa.sub.28-Z.sub.1; wherein Xaa.sub.1 is His, Arg or Tyr or
4-imidazo-propionyl; Xaa.sub.2 is Ser, Gly, Ala or Thr; Xaa.sub.3
is Ala, Asp or Glu; Xaa.sub.5 is Ala or Thr; Xaa.sub.6 is Ala, Phe,
Tyr or naphthylalanine; Xaa.sub.7 is Thr or Ser; Xaa.sub.8 is Ala,
Ser or Thr; Xaa.sub.9 is Asp or Glu; Xaa.sub.10 is Ala, Leu, Ile,
Val, pentylglycine or Met; Xaa.sub.11 is Ala or Ser; Xaa.sub.12 is
Ala or Lys; Xaa.sub.13 is Ala or Gln; Xaa.sub.14 is Ala, Leu, Ile,
pentylglycine, Val or Met; Xaa.sub.15 is Ala or Glu; Xaa.sub.16 is
Ala or Glu; Xaa.sub.17 is Ala or Glu; Xaa.sub.19 is Ala or Val;
Xaa.sub.20 is Ala or Arg; Xaa.sub.21 is Ala, Leu or
Lys-NH.sup..epsilon.--R where R is Lys, Arg, C.sub.1-C.sub.10
straight chain or branched alkanoyl or cycloalkylalkanoyl;
Xaa.sub.22 is Phe, Tyr or naphthylalanine; Xaa.sub.23 is Ile, Val,
Leu, pentylglycine, tert-butylglycine or Met; Xaa.sub.24 is Ala,
Glu or Asp; Xaa.sub.25 is Ala, Trp, Phe, Tyr or naphthylalanine;
Xaa.sub.26 is Ala or Leu; Xaa.sub.27 is Lys, Asn, Ala or
Lys-NH.sup..epsilon.--R where R is Lys, Arg, C.sub.1-C.sub.10
straight chain or branched alkanoyl or cycloalkylalkanoyl; X.sub.28
is Lys, Asn, Ala or Lys-NH.sup..epsilon.--R where R is Lys, Arg,
C.sub.1-C.sub.10 straight chain or branched alkanoyl or
cycloalkylalkanoyl; Z.sub.1 is --OH, --NH.sub.2, Gly-Z.sub.2, Gly
Gly-Z.sub.2, Gly Gly Xaa.sub.31-Z.sub.2, Gly Gly Xaa.sub.31
Ser-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser-Z.sub.2, Gly Gly Xaa.sub.31
Ser Ser Gly-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser Gly Ala-Z.sub.2,
Gly Gly Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36-Z.sub.2, Gly Gly
Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37-Z.sub.2 or Gly Gly
Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37
Xaa.sub.38-Z.sub.2; Xaa.sub.31, Xaa.sub.36, Xaa.sub.37 and
Xaa.sub.38 are independently selected from the group consisting of
Pro, homoproline, 3Hyp, 4Hyp, thioproline, N-alkylglycine,
N-alkylpentylglycine and N-alkylalanine; and Z.sub.2 is --OH or
--NH.sub.2; provided that no more than three of Xaa.sub.3,
Xaa.sub.5, Xaa.sub.6, Xaa.sub.8, Xaa.sub.10, Xaa.sub.11,
Xaa.sub.12, Xaa.sub.13, Xaa.sub.14, Xaa.sub.15, Xaa.sub.16,
Xaa.sub.17, Xaa.sub.19, Xaa.sub.20, Xaa.sub.21, Xaa.sub.24,
Xaa.sub.25, and Xaa.sub.26 are Ala.
[0073] Exemplary exendin analogs of formula (V) include those
wherein Xaa.sub.1 is His, Tyr or 4-imidazopropionyl. More
preferably Xaa.sub.1 is His. Provided are those compounds of
formula (V) wherein Xaa.sub.1 is 4-imidazopropionyl. Provided are
those compounds of formula (V) wherein Xaa.sub.2 is Gly. Exemplary
compounds of formula (V) are those wherein Xaa.sub.14 is Leu,
pentylglycine or Met. Exemplary compounds of formula (V) are those
wherein Xaa.sub.25 is Trp or Phe.
[0074] According to one embodiment, provided are compounds of
formula (V) wherein Xaa.sub.6 is Phe or naphthylalanine; and
Xaa.sub.22 is Phe or naphthylalanine; and Xaa.sub.23 is Ile or Val.
More preferably, Z.sub.1 is --NH.sub.2. According to one
embodiment, provided are compounds of formula (V) wherein
Xaa.sub.31, Xaa.sub.36, Xaa.sub.37 and Xaa.sub.38 are independently
selected from the group consisting of Pro, homoproline, thioproline
and N-alkylalanine. Preferably, Z.sub.2 is --NH.sub.2. Exemplary
compounds of formula (V) include those wherein X.sub.27 is Lys or
Lys-NH.sup..epsilon.--R where R is Lys, Arg, C.sub.1-C.sub.10
straight chain or branched alkanoyl and Xaa.sub.28 is Asn or
Ala.
[0075] Preferred compounds of formula (V) include compounds
described in WO 99/25727 and identified therein as Compound Nos.
62-69.
[0076] Provided herein are exendin analogs wherein Xaa.sub.1 is
His. Provided are those compounds of formula (V) wherein Xaa.sub.2
is Gly. Provided are those compounds of formula (V) wherein
Xaa.sub.3 is Ala. Provided are those compounds of formula (V)
wherein Xaa.sub.14 is Leu, pentylglycine or Met. Provided compounds
of formula (V) are those wherein Xaa.sub.25 is Trp or Phe.
Exemplary compounds of formula (V) are those where Xaa.sub.6 is
Ala, Phe or naphthylalanine; Xaa.sub.22 is Phe or naphthylalanine;
and Xaa.sub.23 is Ile or Val. Provided are compounds of formula (V)
wherein Xaa.sub.31, Xaa.sub.36, Xaa.sub.37 and Xaa.sub.38 are
independently selected from Pro, homoproline, thioproline and
N-alkylalanine. Preferably Z.sub.1 is --NH.sub.2. Preferably
Z.sub.2 is --NH.sub.2.
[0077] According to one embodiment, provided are compounds of
formula (V) wherein Xaa.sub.1 is His or Tyr, more preferably His;
Xaa.sub.2 is Ala or Gly; Xaa.sub.6 is Phe or naphthylalanine;
Xaa.sub.14 is Ala, Leu, pentylglycine or Met; Xaa.sub.22 is Phe or
naphthylalanine; Xaa.sub.23 is Ile or Val; Xaa.sub.31, Xaa.sub.36,
Xaa.sub.37 and Xaa.sub.38 are independently selected from Pro,
homoproline, thioproline or N-alkylalanine; and Xaa.sub.39 is Ser
or Tyr, more preferably Ser. More preferably Z.sub.1 is
--NH.sub.2.
[0078] According to one embodiment, provided compounds include
those of formula (V) wherein: Xaa.sub.1 is His; Xaa.sub.2 is Gly or
Ala; Xaa.sub.3 is Ala, Asp or Glu; Xaa.sub.4 is Gly; Xaa.sub.5 is
Ala or Thr; Xaa.sub.6 is Phe or naphthylalanine; Xaa.sub.7 is Thr
or Ser; Xaa.sub.8 is Ala, Ser or Thr; Xaa.sub.9 is Asp or Glu;
Xaa.sub.10 is Ala, Leu or pentylglycine; Xaa.sub.11 is Ala or Ser;
Xaa.sub.12 is Ala or Lys; Xaa.sub.13 is Ala or Gln; Xaa.sub.14 is
Ala, Leu, Met or pentylglycine; Xaa.sub.15 is Ala or Glu;
Xaa.sub.16 is Ala or Glu; Xaa.sub.17 is Ala or Glu; Xaa.sub.19 is
Ala or Val; Xaa.sub.20 is Ala or Arg; Xaa.sub.21 is Ala or Leu;
Xaa.sub.22 is Phe or naphthylalanine; Xaa.sub.23 is Ile, Val or
tert-butylglycine; Xaa.sub.24 is Ala, Glu or Asp; Xaa.sub.25 is
Ala, Trp or Phe; Xaa.sub.26 is Ala or Leu; Xaa.sub.27 is Ala or
Lys; Xaa.sub.28 is Ala or Asn; Z.sub.1 is --OH, --NH.sub.2,
Gly-Z.sub.2, Gly Gly-Z.sub.2, Gly Gly Xaa.sub.31-Z.sub.2, Gly Gly
Xaa.sub.31 Ser-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser-Z.sub.2, Gly Gly
Xaa.sub.31 Ser Ser Gly-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser Gly
Ala-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36-Z.sub.2,
Gly Gly Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37-Z.sub.2,
Gly Gly Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37
Xaa.sub.38-Z.sub.2 or Gly Gly Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36
Xaa.sub.37 Xaa.sub.38 Xaa.sub.39-Z.sub.2; Xaa.sub.31, Xaa.sub.36,
Xaa.sub.37 and Xaa.sub.38 being independently Pro homoproline,
thioproline or N-methylalanine; and Z.sub.2 being --OH or
--NH.sub.2; provided that no more than three of Xaa.sub.3,
Xaa.sub.5, Xaa.sub.6, Xaa.sub.8, Xaa.sub.10, Xaa.sub.11,
Xaa.sub.12, Xaa.sub.13, Xaa.sub.14, Xaa.sub.15, Xaa.sub.16,
Xaa.sub.17, Xaa.sub.19, Xaa.sub.20, Xaa.sub.21, Xaa.sub.24,
Xaa.sub.25, Xaa.sub.26, Xaa.sub.27 and Xaa.sub.28 are Ala; and
provided also that, if Xaa.sub.1 is His, Arg or Tyr, then at least
one of Xaa.sub.3 and Xaa.sub.4 is Ala.
[0079] According to one embodiment, provided are compounds of
formula (V) where Xaa.sub.14 is Ala, Leu, Ile, Val or
pentylglycine, more preferably Leu or pentylglycine, and Xaa.sub.25
is Ala, Phe, Tyr or naphthylalanine, more preferably Phe or
naphthylalanine. These compounds will be less susceptible to
oxidative degradation, both in vitro and in vivo, as well as during
synthesis of the compound.
[0080] Particular compounds of formula (V) include those described
in WO 99/25727 and having the amino acid sequences identified
therein as SEQ ID NOS: 5-65 and 67-74, herein SEQ ID NOs:
23-91.
[0081] Also provided for use in the formulation and methods
described herein are peptides described in U.S. Pat. No. 7,220,721,
including compounds of formula (VI) (SEQ ID NO:20) or a
pharmaceutically acceptable salt thereof: Xaa.sub.1 Xaa.sub.2
Xaa.sub.3 Xaa.sub.4 Xaa.sub.5 Xaa.sub.6 Xaa.sub.7 Xaa.sub.8
Xaa.sub.9 Xaa.sub.10 Xaa.sub.11 Xaa.sub.12 Xaa.sub.13 Xaa.sub.14
Xaa.sub.15 Xaa.sub.16 Xaa.sub.17 Ala Xaa.sub.19 Xaa.sub.20
Xaa.sub.21 Xaa.sub.22 Xaa.sub.23 Xaa.sub.24 Xaa.sub.25 Xaa.sub.26
Xaa.sub.27 Xaa.sub.28-Z.sub.1; wherein Xaa.sub.1 is His, Arg, Tyr,
Ala, Norval, Val, Norleu or 4-imidazopropionyl; Xaa.sub.2 is Ser,
Gly, Ala or Thr; Xaa.sub.3 is Ala, Asp or Glu; Xaa.sub.4 is Ala,
Norval, Val, Norleu or Gly; Xaa.sub.5 is Ala or Thr; Xaa.sub.6 is
Ala, Phe, Tyr or naphthylalanine; Xaa.sub.7 is Thr or Ser;
Xaa.sub.8 is Ala, Ser or Thr; Xaa.sub.9 is Ala, Norval, Val,
Norleu, Asp or Glu; Xaa.sub.10 is Ala, Leu, Ile, Val, pentylglycine
or Met; Xaa.sub.11 is Ala or Ser; Xaa.sub.12 is Ala or Lys;
Xaa.sub.13 is Ala or Gln; Xaa.sub.14 is Ala, Leu, Ile,
pentylglycine, Val or Met; Xaa.sub.15 is Ala or Glu; Xaa.sub.16 is
Ala or Glu; Xaa.sub.17 is Ala or Glu; Xaa.sub.19 is Ala or Val;
Xaa.sub.20 is Ala or Arg; Xaa.sub.21 is Ala, Leu or
Lys-NH.sup..epsilon.--R where R is Lys, Arg, C.sub.1-10 straight
chain or branched alkanoyl or cycloalkyl-alkanoyl; Xaa.sub.22 is
Phe, Tyr or naphthylalanine; Xaa.sub.23 is Ile, Val, Leu,
pentylglycine, tert-butylglycine or Met; Xaa.sub.24 is Ala, Glu or
Asp; Xaa.sub.25 is Ala, Trp, Phe, Tyr or naphthylalanine;
Xaa.sub.26 is Ala or Leu; Xaa.sub.27 is Lys, Asn,
Lys-NH.sup..epsilon.--R or Ala where R is Lys, Arg,
C.sub.1-C.sub.10 straight chain or branched alkanoyl or
cyclo-alkylalkanoyl; Xaa.sub.28 is Lys, Asn,
Lys-NH.sup..epsilon.--R or Ala where R is Lys, Arg,
C.sub.1-C.sub.10 straight chain or branched alkanoyl or
cycloalkylalkanoyl; Z.sub.1 is --OH, --NH.sub.2, Gly-Z.sub.2, Gly
Gly-Z.sub.2, Gly Gly Xaa.sub.31-Z.sub.2, Gly Gly Xaa.sub.31
Ser-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser-Z.sub.2, Gly Gly Xaa.sub.31
Ser Ser Gly-Z.sub.2, Gly Gly Xaa.sub.31 Ser Ser Gly Ala-Z.sub.2,
Gly Gly Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36-Z.sub.2, Gly Gly
Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37-Z.sub.2, Gly Gly
Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37 Xaa.sub.38-Z.sub.2
or Gly Gly Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37
Xaa.sub.38 Xaa.sub.39-Z.sub.2; Xaa.sub.31, Xaa.sub.36, Xaa.sub.37
and Xaa.sub.38 are independently selected from the group consisting
of Pro, homoproline, 3Hyp, 4Hyp, thioproline, N-alkylglycine,
N-alkylpentylglycine and N-alkylalanine; Xaa.sub.39 is Ser or Tyr;
and Z.sub.2 is --OH or --NH.sub.2; provided that no more than three
of Xaa.sub.3, Xaa.sub.4, Xaa.sub.5, Xaa.sub.6, Xaa.sub.8,
Xaa.sub.9, Xaa.sub.10, Xaa.sub.11, Xaa.sub.12, Xaa.sub.13,
Xaa.sub.14, Xaa.sub.15, Xaa.sub.16, Xaa.sub.17, Xaa.sub.19,
Xaa.sub.20, Xaa.sub.21, Xaa.sub.24, Xaa.sub.25, Xaa.sub.26, are
Ala; and provided also that, if Xaa.sub.1 is His, Arg, Tyr, or
4-imidazopropionyl then at least one of Xaa.sub.3, Xaa.sub.4 and
Xaa.sub.9 is Ala.
[0082] Exemplary compounds of formula (VI) include those wherein
Xaa.sub.1 is His, Ala, Norval or 4-imidazopropionyl. Preferably,
Xaa.sub.1 is His, or 4-imidazopropionyl or Ala, more preferably His
or 4-imidazopropionyl. Exemplary compounds of formula (VI) include
those wherein Xaa.sub.2 is Gly. Exemplary compounds of formula (VI)
include those wherein Xaa.sub.4 is Ala. Exemplary compounds of
formula (VI) include those wherein Xaa.sub.9 is Ala. Exemplary
compounds of formula (VI) include those wherein Xaa.sub.14 is Leu,
pentylglycine or Met. Exemplary compounds of formula (VI) include
those wherein Xaa.sub.25 is Trp or Phe. Exemplary compounds of
formula (VI) include those wherein Xaa.sub.6 is Ala, Phe or
naphthylalanine; Xaa.sub.22 is Phe or naphthylalanine; and
Xaa.sub.23 is Ile or Val. Exemplary compounds of formula (VI)
include those wherein Z.sub.1 is --NH.sub.2. Exemplary compounds of
formula (VI) include those wherein Xaa.sub.31, Xaa.sub.36,
Xaa.sub.37 and Xaa.sub.38 are independently selected from the group
consisting of Pro, homoproline, thioproline and N-alkylalanine.
Exemplary compounds of formula (VI) include those wherein
Xaa.sub.39 is Ser. Exemplary compounds of formula (VI) include
those wherein Z.sub.2 is --NH.sub.2. Exemplary compounds of formula
(VI) include those wherein Z.sub.1 is --NH.sub.2. Exemplary
compounds of formula (VI) include those wherein Xaa.sub.21 is
Lys-NH.sup..epsilon.--R where R is Lys, Arg, C.sub.1-C.sub.10
straight chain or branched alkanoyl. Exemplary compounds of formula
(VI) include those wherein X.sub.27 is Lys or
Lys-NH.sup..epsilon.--R, where R is Lys, Arg, C.sub.1-C.sub.10
straight chain or branched alkanoyl and X.sub.28 is Asn or Ala.
[0083] Other compounds of formula (VI) include those described in
WO 99/25728 as having an amino acid sequence selected from those
identified therein as SEQ ID NOS: 95-110, and herein identified as
SEQ ID NOS: 92-107.
[0084] Compounds useful according to the formulations and methods
described herein are exendin analog agonists described in U.S.
Patent No. 6,956,026, including compounds of formula (VII) (SEQ ID
NO:21) or a pharmaceutically acceptable salt thereof: Xaa.sub.1
Xaa.sub.2 Xaa.sub.3 Gly Thr Xaa.sub.6 Xaa.sub.7 Xaa.sub.8 Xaa.sub.9
Xaa.sub.10 Ser Lys Gln Xaa.sub.14 Glu Glu Glu Ala Val Arg Leu
Xaa.sub.22 Xaa.sub.23 Xaa.sub.24 Xaa.sub.25 Leu Lys Asn Gly Gly
Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37 Xaa.sub.38
Xaa.sub.39-Z; wherein: Xaa.sub.1 is His, Arg or Tyr; Xaa.sub.2 is
Ser, Gly, Ala or Thr; Xaa.sub.3 is Asp or Glu; Xaa.sub.6 is Phe,
Tyr or naphthylalanine; Xaa.sub.7 is Thr or Ser; Xaa.sub.8 is Ser
or Thr; Xaa.sub.9 is Asp or Glu; Xaa.sub.10 is Leu, Ile, Val,
pentylglycine or Met; Xaa.sub.14 is Leu, Ile, pentylglycine, Val or
Met; Xaa.sub.22 is Phe, Tyr or naphthylalanine; Xaa.sub.23 is Ile,
Val, Leu, pentylglycine, tert-butylglycine or Met; Xaa.sub.24 is
Glu or Asp; Xaa.sub.25 is Trp, Phe, Tyr, or naphthylalanine;
Xaa.sub.31, Xaa.sub.36, Xaa.sub.37 and Xaa.sub.38 are independently
Pro, homoproline, 3Hyp, 4Hyp, thioproline, N-alkylglycine,
N-alkylpentylglycine or N-alkyl-alanine; Xaa.sub.39 is Ser, Thr or
Tyr; and Z is --OH or --NH.sub.2; with the proviso that the
compound does not have the formula of either SEQ ID NOS:1 or 2.
[0085] Preferred N-alkyl groups for N-alkylglycine,
N-alkylpentylglycine and N-alkylalanine include lower alkyl groups
preferably of 1 to about 6 carbon atoms, more preferably of 1 to 4
carbon atoms. Exemplary exendin analogs include those wherein
Xaa.sub.1 is His or Tyr. More preferably Xaa.sub.1 is His. Provided
are those compounds wherein Xaa.sub.2 is Gly. Provided are those
compounds wherein Xaa.sub.14 is Leu, pentylglycine or Met.
Exemplary compounds include those wherein Xaa.sub.25 is Trp or Phe.
Also provided are compounds where Xaa.sub.6 is Phe or
naphthylalanine; Xaa.sub.23 is Ile or Val and Xaa.sub.31,
Xaa.sub.36, Xaa.sub.37 and Xaa.sub.38 are independently selected
from Pro, homoproline, thioproline or N-alkylalanine. Preferably
N-alkylalanine has a N-alkyl group of 1 to about 6 carbon atoms.
According to one embodiment, Xaa.sub.36, Xaa.sub.37 and Xaa.sub.38
are the same amino acid reside. Provided are compounds wherein
Xaa.sub.39 is Ser or Tyr, more preferably Ser. Preferably Z is
--NH.sub.2.
[0086] According to one embodiment, provided are compounds of
formula (VII) wherein Xaa.sub.1 is His or Tyr, preferably His;
Xaa.sub.2 is Gly; Xaa.sub.6 is Phe or naphthylalanine; Xaa.sub.14
is Leu, pentylglycine or Met; Xaa.sub.22 is Phe or naphthylalanine;
Xaa.sub.23 is Ile or Val; Xaa.sub.31, Xaa.sub.36, Xaa.sub.37 and
Xaa.sub.38 are independently selected from Pro, homoproline,
thioproline or N-alkylalanine; and Xaa.sub.39 is Ser or Tyr, more
preferably Ser. More preferably Z is --NH.sub.2.
[0087] According to another embodiment, exemplary compounds include
those of formula (VII) wherein: Xaa.sub.1 is His or Arg; Xaa.sub.2
is Gly; Xaa.sub.3 is Asp or Glu; Xaa.sub.6 is Phe or
napthylalanine; Xaa.sub.7 is Thr or Ser; Xaa.sub.8 is Ser or Thr;
Xaa.sub.9 is Asp or Glu; Xaa.sub.10 is Leu or pentylglycine;
Xaa.sub.14 is Leu or pentylglycine; Xaa.sub.22 is Phe or
naphthylalanine; Xaa.sub.23 is Ile, Val or t-butylglycine;
Xaa.sub.24 is Glu or Asp; Xaa.sub.25 is Trp or Phe; Xaa.sub.31,
Xaa.sub.36, Xaa.sub.37, and Xaa.sub.38 are independently Pro,
homoproline, thioproline, or N-methylalanine; Xaa.sub.39 is Ser or
Tyr: and Z is --OH or --NH.sub.2; with the proviso that the
compound does not have the formula of either SEQ ID NOS:1 or 2.
More preferably Z is --NH.sub.2.
[0088] According to another embodiment, provided are compounds
where Xaa.sub.14 is Leu, Ile, Val or pentylglycine, more preferably
Leu or pentylglycine, and Xaa.sub.25 is Phe, Tyr or
naphthylalanine, more preferably Phe or naphthylalanine. These
compounds are believed to exhibit advantageous duration of action
and to be less patient to oxidative degradation, both in vitro and
in vivo, as well as during synthesis of the compound.
[0089] Also provided for use in the formulations and methods
described herein are compounds described in U.S. Pat. No.
7,157,555, including compounds of formula (VIII) (SEQ ID NO:22) or
a pharmaceutically acceptable salt thereof: Xaa.sub.1 Xaa.sub.2
Xaa.sub.3 Gly Thr Xaa.sub.6 Xaa.sub.7 Xaa.sub.8 Xaa.sub.9
Xaa.sub.10 Ser Lys Gln Xaa.sub.14 Glu Glu Glu Ala Val Arg Leu
Xaa.sub.22 Xaa.sub.23 Xaa.sub.24 Xaa.sub.25 Leu Xaa.sub.27
Xaa.sub.28 Gly Gly Xaa.sub.31 Ser Ser Gly Ala Xaa.sub.36 Xaa.sub.37
Xaa.sub.38 Xaa.sub.39-Z; wherein: Xaa.sub.1 is His, Arg, Tyr or
4-imidazopropionyl; Xaa.sub.2 is Ser, Gly, Ala or Thr; Xaa.sub.3 is
Asp or Glu; Xaa.sub.6 is Phe, Tyr or naphthylalanine; Xaa.sub.7 is
Thr or Ser; Xaa.sub.8 is Ser or Thr; Xaa.sub.9 is Asp or Glu;
Xaa.sub.10 is Leu, Ile, Val, pentylglycine or Met; Xaa.sub.14 is
Leu, Ile, pentylglycine, Val or Met; Xaa.sub.22 is Phe, Tyr or
naphthylalanine; Xaa.sub.23 is Ile, Val, Leu, pentylglycine,
tert-butylglycine or Met; Xaa.sub.24 is Glu or Asp; Xaa.sub.25 is
Trp, Phe, Tyr, or naphthylalanine; Xaa.sub.27 is Lys, Asn, or
Lys-NH.sup..epsilon.--R where R is Lys, Arg, C.sub.1-C.sub.10
straight chain or branched alkanoyl or cycloalkylalkanoyl;
Xaa.sub.28 is Lys, Asn, or Lys-NH.sup..epsilon.--R where R is Lys,
Arg, C.sub.1-C.sub.10 straight chain or branched alkanoyl or
cycloalkylalkanoyl; Xaa.sub.31, Xaa.sub.36, Xaa.sub.37 and
Xaa.sub.38 are independently Pro, homoproline, 3Hyp, 4Hyp,
thioproline, N-alkylglycine, N-alkylpentylglycine or
N-alkylalanine; Xaa.sub.39 is Ser, Thr or Tyr; and Z is --OH or
--NH.sub.2; with the proviso that the compound does not have the
formula of either SEQ ID NOS: 1 or 2.
[0090] Exemplary exendin analogs of formula (VIII) include those
wherein Xaa.sub.1 is His, Tyr or 4-imidazopropionyl. More
preferably, Xaa.sub.1 is His or 4-imidazopropionyl. Provided are
those compounds of formula (VIII) wherein Xaa.sub.2 is Gly.
Provided are those compounds of formula (VIII) wherein Xaa.sub.14
is Leu, pentylglycine or Met. Provided are those compounds of
formula (VIII) wherein Xaa.sub.25 is Trp or Phe. Provided are those
compounds of formula (VIII) wherein Xaa.sub.27 is Lys or
Lys-NH.sup..epsilon.--R where R is Lys, Arg, C.sub.1-C.sub.10
straight chain or branched alkanoyl and Xaa.sub.28 is Asn.
[0091] Also provided are compounds of formula (VIII) wherein
Xaa.sub.6 is Phe or naphthylalanine; Xaa.sub.22 is Phe or
naphthylalanine; Xaa.sub.23 is Ile or Val and Xaa.sub.31,
Xaa.sub.36, Xaa.sub.37 and Xaa.sub.38 are independently selected
from Pro, homoproline, thioproline or N-alkylalanine. According to
one embodiment, Xaa.sub.39 is Ser or Tyr. Provide are compounds
wherein Xaa.sub.39 is Ser. Preferably, Z is --NH.sub.2.
[0092] According to one embodiment, provided are compounds of
formula (VIII) wherein Xaa.sub.6 is Phe or naphthylalanine;
Xaa.sub.22 is Phe or naphthylalanine; Xaa.sub.23 is Ile or Val;
Xaa.sub.27 is Lys or Lys-NH.sup..epsilon.--R where R is Lys, Arg,
C.sub.1-C.sub.10 straight chain or branched alkanoyl, and
Xaa.sub.28 is Asn; and Xaa.sub.31, Xaa.sub.36, Xaa.sub.37 and
Xaa.sub.38 are independently selected from Pro, homoproline,
thioproline or N-alkylalanine.
[0093] In another embodiment, exendins and exendin analogs of the
invention do not include the peptides of SEQ ID NOS:3-14. In one
embodiment, exendin analogs include the analogs of Formulas
(I-VIII), with the proviso that the analogs do not include the
peptides of SEQ ID NOs: 1-2.
[0094] Also useful within the scope of the invention are narrower
genera of compounds of the described formulae, for example formulae
I through VIII, having peptides of various lengths, for example
genera of compounds that do not include peptides having a length of
greater than 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37 or 38 amino
acid residues.
[0095] The exendins, exendin agonists, or exendin analog agonists
described herein may be in the form of a peptide, a pro-drug, or as
a pharmaceutical salt or salts thereof. The term "prodrug" refers
to a compound that is a drug precursor that, following
administration, releases the drug in vivo via some chemical or
physiological process, for example, proteolytic cleavage, or upon
reaching an environment of a certain pH.
[0096] Exendins, exendin analog agonists and exendin agonists that
are peptides, described herein may be prepared through peptide
purification as described in, for example, Eng et al, J. Biol.
Chem., 265:20259-62 (1990); and Eng et al, J. Biol. Chem.,
267:7402-05 (1992), which are incorporated by reference herein.
Alternatively, exendins, exendin peptide agonists and exendin
analog agonists may be prepared by methods known to those skilled
in the art, for example, as described in Raufman et al, J. Biol.
Chem., 267:21432-37 (1992), which is incorporated by reference
herein, using standard solid-phase peptide synthesis techniques,
for example, using an automated or semiautomated peptide
synthesizer. Typically, using such techniques, an u-N-carbamoyl
protected amino acid and an amino acid attached to the growing
peptide chain on a resin are coupled at room temperature in an
inert solvent such as dimethylformamide, N-methylpyrrolidinone or
methylene chloride in the presence of coupling agents such as
dicyclohexylcarbodiimide and 1-hydroxybenzotriazole in the presence
of a base such as diisopropylethylamine. The .alpha.-N-carbamoyl
protecting group is removed from the resulting peptide-resin using
a reagent such as trifluoroacetic acid or piperidine, and the
coupling reaction repeated with the next desired N-protected amino
acid to be added to the peptide chain. Suitable N-protecting groups
are well known in the art, with, for example, t-butyloxycarbonyl
(tBoc) and fluorenylmethoxycarbonyl (Fmoc).
[0097] The solvents, amino acid derivatives and
4-methylbenzhydryl-amine resin used in the peptide synthesizer may
be purchased from Applied Biosystems Inc. (Foster City, Calif.).
The following side-chain protected amino acids may be purchased
from Applied Biosystems, Inc.: BSD-112344.1-Arg(Pmc), Boc-Thr(Bzl),
Fmoc-Thr(t-Bu), Boc-Ser(Bzl), Fmoc-Ser(t-Bu), Boc-Tyr(BrZ),
Fmoc-Tyr(t-Bu), Boc-Lys(Cl-Z), Fmoc-Lys(Boc), Boc-Glu(Bzl),
Fmoc-Glu(t-Bu), Fmoc-His(Trt), Fmoc-Asn(Trt), and Fmoc-Gln(Trt).
Boc-His(BOM) may be purchased from Applied Biosystems, Inc. or
Bachem Inc. (Torrance, Calif.). Anisole, dimethylsulfide, phenol,
ethanedithiol, and thioanisole may be obtained from Aldrich
Chemical Company (Milwaukee, Wis.). Air Products and Chemicals
(Allentown, Pa.) supplies HF. Ethyl ether, acetic acid and methanol
may be purchased from Fisher Scientific (Pittsburgh, Pa.).
[0098] Solid phase peptide synthesis may be carried out with an
automatic peptide synthesizer (Model 430A, Applied Biosystems Inc.,
Foster City, Calif.) using the NMP/HOBt (Option 1) system and tBoc
or Fmoc chemistry (see, Applied Biosystems User's Manual for the
ABI 430A Peptide Synthesizer, Version 1.3B Jul. 1, 1988, section 6,
pp. 49-70, Applied Biosystems, Inc., Foster City, Calif.) with
capping. Boc-peptide-resins may be cleaved with HF (-50.degree. C.
to 0.degree. C., 1 hour). The peptide may be extracted from the
resin with alternating water and acetic acid, and the filtrates
lyophilized. The Fmoc-peptide resins may be cleaved according to
standard methods (Introduction to Cleavage Techniques, Applied
Biosystems, Inc., 1990, pp. 6-12). Peptides may also be assembled
using an Advanced Chem Tech Synthesizer (Model MPS 350, Louisville,
Ky.).
[0099] Peptides may be purified by RP-HPLC (preparative and
analytical) using a Waters Delta Prep 3000 system. A C4, C8 or C18
preparative column (10.mu., 2.2.times.25 cm; Vydac, Hesperia,
Calif.) may be used to isolate peptides, and purity may be
determined using a C4, C8 or C18 analytical column (5.mu.,
0.46.times.25 cm; Vydac). Solvents (A=0.1% TFA/water and B=0.1%
TFA/CH.sub.3CN) may be delivered to the analytical column at a
flowrate of 1.0 ml/min and to the preparative column at 15 ml/min.
Amino acid analyses may be performed on the Waters Pico Tag system
and processed using the Maxima program. Peptides may be hydrolyzed
by vapor-phase acid hydrolysis (115.degree. C., 20-24 h).
Hydrolysates may be derivatized and analyzed by standard methods
(Cohen, et al., The Pico Tag Method: A Manual of Advanced
Techniques for Amino Acid Analysis, pp. 11-52, Millipore
Corporation, Milford, Mass. (1989)). Fast atom bombardment analysis
may be carried out by M-Scan, Incorporated (West Chester, Pa.).
Mass calibration may be performed using cesium iodide or cesium
iodide/glycerol. Plasma desorption ionization analysis using time
of flight detection may be carried out on an Applied Biosystems
Bio-Ion 20 mass spectrometer. Electrospray mass spectroscopy may be
carried and on a VG-Trio machine.
[0100] Exendins, exendin analog agonists and exendin agonists that
are peptides may also be prepared using recombinant DNA techniques,
using methods now known in the art. See, e.g., Sambrook et al,
Molecular Cloning: A Laboratory Manual, 2d Ed., Cold Spring Harbor
(1989). Alternatively, such compounds may be prepared by
homogeneous phase peptide synthesis methods. Non-peptide compounds
useful in the invention may be prepared by art-known methods. For
example, phosphate-containing amino acids and peptides containing
such amino acids, may be prepared using methods known in the art.
See, e.g., Bartlett and Landen, Biorg. Chem., 14:356-377
(1986).
[0101] Exendins, exendin agonists or exendin analog agonists may be
formulated into pharmaceutical compositions for administration to
patients. These pharmaceutical compositions preferably include an
amount of an exendin, an exendin agonist or exendin analog agonist
effective to lower total cholesterol, lower LDL cholesterol, lower
triglyceride levels, or treat dyslipidemia or atherosclerosis, and
a pharmaceutically acceptable carrier.
[0102] In any of the embodiments described herein, the patient may
be a mammal, including a human or an animal. The animal may be a
domestic animal, such as a companion animal (e.g., dog, cat, horse)
or livestock (e.g., sheep, cow, pig, buffalo, ostrich, chicken,
turkey).
[0103] The compositions described herein may be administered
parenterally, orally, by inhalation spray, topically, rectally,
nasally, buccally, vaginally or via an implanted reservoir. The
term "parenteral" as used herein includes subcutaneous,
intravenous, intramuscular, intra-articular, intra-synovial,
intrasternal, intrathecal, intrahepatic, intralesional and
intracranial injection or infusion techniques. In one embodiment,
the compositions are administered by an infusion pump or
subcutaneous injection of a slow release, extended release,
sustained release or long acting formulation. In one embodiment,
subcutaneous injections are administered once a day; once every
two, three, four, five, or six days; once per week; twice per
month; once a month; every other month or every third month.
[0104] Any of the exendins, exendin agonists or exendin analog
agonists may be administered in the acid or amide form.
Additionally, any of the exendins, exendin agonists or exendin
analog agonists may form salts with various inorganic and organic
acids and bases. Such salts include, without limitation, salts
prepared with organic and inorganic acids, for example, HCl, HBr,
H.sub.2SO.sub.4, H.sub.3PO.sub.4, trifluoroacetic acid, acetic
acid, formic acid, methanesulfonic acid, toluenesulfonic acid,
maleic acid, fumaric acid and camphorsulfonic acid. Salts prepared
with bases include, without limitation, ammonium salts, alkali
metal salts, e.g., sodium and potassium salts, and alkali earth
salts, e.g., calcium and magnesium salts. Acetate, hydrochloride,
and trifluoroacetate salts are particular examples. The salts may
be formed by conventional means, as by reacting the free acid or
base forms of the product with one or more equivalents of the
appropriate base or acid in a solvent or medium in which the salt
is insoluble, or in a solvent such as water which is then removed
in vacuo or by freeze-drying or by exchanging the ions of an
existing salt for another ion on a suitable ion exchange resin.
[0105] Pharmaceutically acceptable carriers useful in these
pharmaceutical compositions include, e.g., ion exchangers, alumina,
aluminum stearate, lecithin, serum proteins, such as human serum
albumin, buffer substances such as phosphates, glycine, sorbic
acid, potassium sorbate, partial glyceride mixtures of saturated
vegetable fatty acids, water, salts or electrolytes, such as
protamine sulfate, disodium hydrogen phosphate, potassium hydrogen
phosphate, sodium chloride, zinc salts, colloidal silica, magnesium
trisilicate, polyvinyl pyrrolidone, cellulose-based substances,
polyethylene glycol, sodium carboxymethylcellulose, polyacrylates,
waxes, polyethylene-polyoxypropylene-block polymers, polyethylene
glycol and wool fat.
[0106] Sterile injectable forms of the compositions of this
invention may be aqueous or oleaginous suspension. These
suspensions may be formulated according to techniques known in the
art, using suitable dispersing or wetting agents and suspending
agents. The sterile injectable preparation may also be a sterile
injectable solution or suspension in a non-toxic parenterally
acceptable diluent or solvent, for example as a solution in
1,3-butanediol. Among the acceptable vehicles and solvents that may
be employed are water, Ringer's solution and isotonic sodium
chloride solution. In addition, sterile, fixed oils are
conventionally employed as a solvent or suspending medium. For this
purpose, any bland fixed oil may be employed including synthetic
mono- or di-glycerides. Fatty acids, such as oleic acid and its
glyceride derivatives are useful in the preparation of injectables,
as are natural pharmaceutically-acceptable oils, such as olive oil
or castor oil, especially in their polyoxyethylated versions. These
oil solutions or suspensions may also contain a long-chain alcohol
diluent or dispersant, such as carboxymethyl cellulose or similar
dispersing agents that are commonly used in the formulation of
pharmaceutically acceptable dosage forms, including emulsions and
suspensions. Other commonly used surfactants, such as TWEENs, Spans
and other emulsifying agents or bioavailability enhancers which are
commonly used in the manufacture of pharmaceutically acceptable
solid, liquid, or other dosage forms may also be used for the
purposes of formulation.
[0107] Parenteral formulations may be a single bolus dose, an
infusion, or a loading bolus dose followed with a maintenance dose.
These compositions may be administered according to any dosage
schedule described herein.
[0108] The pharmaceutical compositions of this invention may be
orally administered in any orally acceptable dosage form including,
capsules, tablets, aqueous suspensions or solutions. In the case of
tablets for oral use, carriers commonly used include lactose and
corn starch. Lubricating agents, such as magnesium stearate, are
also typically added. For oral administration in a capsule form,
useful diluents include lactose and dried cornstarch. When aqueous
suspensions are required for oral use, the active ingredient is
combined with emulsifying and suspending agents. If desired,
certain sweetening, flavoring or coloring agents may also be
added.
[0109] The pharmaceutical compositions of this invention may also
be administered topically. For topical applications, the
pharmaceutical compositions may be formulated in a suitable
ointment containing the active component suspended or dissolved in
one or more carriers. Carriers for topical administration of the
compounds of this invention include, mineral oil, liquid
petrolatum, white petrolatum, propylene glycol, polyoxyethylene,
polyoxypropylene compound, emulsifying wax and water.
Alternatively, the pharmaceutical compositions can be formulated in
a suitable lotion or cream containing the active components
suspended or dissolved in one or more pharmaceutically acceptable
carriers. Suitable carriers include, but are not limited to,
mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters
wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol, and
water.
[0110] The pharmaceutical compositions of this invention may also
be administered by nasal aerosol or inhalation. Such compositions
are prepared according to techniques well-known in the art of
pharmaceutical formulation and may be prepared as solutions in
saline, employing benzyl alcohol or other suitable preservatives,
absorption promoters to enhance bioavailability, fluorocarbons,
and/or other conventional solubilizing or dispersing agents.
[0111] The amount of exendin, exendin agonist or exendin analog
agonist that may be combined with the carrier materials to produce
a single dosage form will vary depending upon the host treated and
the particular mode of administration. The compositions can be
formulated so that a dosage of between 0.1-1000 pmoles/kg body
weight/minute (when administered by infusion) of exendin, exendin
agonists or exendin analog agonist is administered to a patient
receiving these compositions. In some embodiments of the invention,
the dosage is 1-10 pmoles/kg body weight/minute (when administered
by infusion). In one embodiment the dosage is 0.5-2.0 pmoles/kg/min
when administered by intravenous infusion.
[0112] The pharmaceutical formulations comprising the exendin,
exendin agonist or exendin analog agonist may be administered as a
single dose or multiple doses. In embodiment, the pharmaceutical
formulation dosage is about 0.8 mg containing about 5% of an
exendin, exendin agonist, or exendin analog agonist, such as, for
example, exendin-4. In another embodiment, the pharmaceutical
formulation dosage is about 2.0 mg containing about 5% of an
exendin, exendin agonist, or exendin analog agonist, such as, for
example, exendin-4. In additional embodiments, the pharmaceutical
formulation dosage is about 0.5 mg, about 0.8 mg, about 1.0 mg,
about 1.25 mg, about 1.5 mg, about 1.75 mg, about 2.0 mg, about
2.25 mg, about 2.5 mg, about 2.75 mg, about 3.0 mg, about 3.25 mg,
about 3.5 mg, about 3.75 mg, about 4.0 mg, about 4.25 mg, about 4.5
mg, about 4.75 mg, about 5.0 mg, or any range between a lower
amount and a higher amount within these described dosages (e.g.,
about 0.5 mg to about 5.0 mg; about 1.75 mg to about 2.25 mg, and
the like) where the pharmaceutical formulation dosage contains from
about 1% to about 20%, from about 2% to about 10%, about 3% to
about 7%; or about 5% of an exendin, exendin agonist, or exendin
analog agonist, such as, for example, exendin-4. The skilled
artisan can make any necessary dosage adjustments in order to
obtain the effective plasma concentrations of an exendin, exendin
agonist or exendin analog agonist suitable for use in the methods
described herein.
[0113] In other embodiments, the pharmaceutical formulations
described herein may contain from about from about 20 .mu.g to
about 400 .mu.g; from about 30 .mu.g to about 300 .mu.g; or from
about 50 .mu.g to about 200 .mu.g of an exendin, exendin agonist,
or exendin analog agonist. In one embodiment, the pharmaceutical
formulation is in a form that can be administered subcutaneously.
In another embodiment, the pharmaceutical formulation is a
polymer-based formulation that can be administered subcutaneously.
The skilled artisan can make any necessary adjustments to the
amount of the exendin, exendin agonist, or exendin analog agonist
used in the pharmaceutical formulations in order to obtain the
effective plasma concentrations of the exendin, exendin agonist or
exendin analog agonist suitable for use in the methods described
herein.
[0114] In another embodiment, the exendins exendin agonists or
exendin analog agonists are formulated in a sustained release or
long acting formulation. In one embodiment, the sustained release
formulation comprises a biocompatible polymer, an exendin, or
exendin analog agonist, and a sugar. Exemplary formulations are
described in U.S. Pat. No. 6,824,822; US Publication No.
2006/0099271; US Publication No. 2004/0228833; US Publication No.
2004/0208929; US Publication No. 2005/0271702; and US Publication
No. 2006/0110423, the disclosures of which are incorporated by
reference herein in their entirety. Such formulations can be
administered, for example by subcutaneous injection, once per day,
every 2 days, 3 days, 4 days, 5 days, or 6 days; once per week,
once every 2 weeks, or once every three weeks; or once per month,
once every other month or once every 3 months.
[0115] Sustained release compositions can be prepared by a phase
separation process. The general process for producing a sustained
release or long acting formulation comprising microparticles
containing an exendin, exendin agonist or exendin analog agonist
and sucrose for a 1 kg batch size is described below.
[0116] A water-in-oil emulsion is created with the aid of a
homogenizer. Suitable homogenizers include an in-line Megatron
homogenizer MT-V 3-65 F/FF/FF, Kinematica AG, Switzerland. The
water phase of the emulsion can be prepared by dissolving an
exendin, exendin agonist or exendin analog agonist, for example,
exendin-4, and excipients such as sucrose in water. The
concentration of exendin in the resulting solution can be from
about 50 mg/g to about 100 mg/g. For example, when the drug is
exendin-4, the concentration of drug in solution can be from about
30 g to about 60 g per 600 g of water. In a particular embodiment,
50 g exendin-4 and 20 g sucrose are dissolved in 600 g water for
irrigation (WFI). The specified amounts listed above represent a
nominal load without adjustment to compensate for peptide content
strength specific to the lot of exendin-4 used. The oil phase of
the emulsion is prepared by dissolving PLGA polymer (e.g., 930 g of
purified 50:50 DL4A PLGA (Alkermes, Inc.) in methylene chloride
(14.6 kg or 6% w/w)).
[0117] The water phase is then added to the oil phase to form a
coarse emulsion with an overhead mixer for about three minutes.
Then, the coarse emulsion is homogenized at approximately 21300 rpm
at ambient temperature for three discrete periods. This should
result in an inner emulsion droplet size of less than 1 micron. It
is understood that inner emulsion formation can be achieved using
any suitable means. Suitable means of emulsion formation include,
but are not limited to, homogenization as described above and
sonication.
[0118] A coacervation step is then performed by adding silicone oil
(21.8 kg of Dimethicone, NF, 350 cs) over a time period of less
than or equal to about 5 minutes to the inner emulsion. This is
equivalent to a ratio of 1.5:1, silicone oil to methylene chloride.
The methylene chloride from the polymer solution partitions into
the silicone oil and begins to precipitate the polymer around the
water phase containing the exendin, leading to microencapsulation.
The embryonic microspheres thus formed are soft and require
hardening. Frequently, the embryonic microspheres are permitted to
stand for a short period of time, for example, less than 1 minute
or from about 1 minute to about 5 minutes prior to proceeding to
the microsphere hardening step.
[0119] The embryonic microspheres are immediately transferred into
a heptane/ethanol solvent mixture. The volume of heptane/ethanol
mixture needed can be determined based on the microsphere batch
size, typically a 16:1 ratio of heptane/ethanol solvent to
methylene chloride. For example, about 210 kg heptane and 23 kg
ethanol in a 3.degree. C. cooled, stirred tank can be used. This
solvent mixture hardens the microspheres by extracting additional
methylene chloride from the microspheres. This hardening step can
also be referred to as quenching. After being quenched for 1 hour
at 3.degree. C., the solvent mixture is either decanted and fresh
heptane (13 Kg) is added at 3.degree. C. and held for 1 hour to
rinse off residual silicone oil, ethanol and methylene chloride on
the microsphere surface or pumped directly to the collection
step.
[0120] At the end of the quench or decant/wash step, the
microspheres are transferred and collected, for example, on a 12''
Sweco Pharmasep Filter/Dryer Model PH12Y6. In this example, the
filter/dryer uses a 25 micron multilayered collection screen and is
connected to a motor that vibrates the screen during collection and
drying. A final rinse with heptane (6 Kg at 3.degree. C.) can be
performed to ensure maximum line transfer and to remove any excess
silicone oil. The microspheres can then be dried under vacuum with
or without a constant purge of nitrogen gas at controlled rate, for
example: 3 to 10 hours (e.g. 6 hours) at 3.degree. C.; 3 to 10
hours ramping to 41.degree. C. (e.g. 6 hours); and maintaining for
a long period (e.g. 80-90 hours) at 41.degree. C. After the
completion of drying, the microspheres are discharged into a
collection vessel, sieved through a 150 .mu.m sieve, and stored at
about -20.degree. C. until filling.
[0121] An alternative general process for producing a sustained
release or long acting formulation comprising microparticles
containing an exendin, exendin agonist or exendin analog agonist
and sucrose is as follows: A water-in-oil emulsion is created with
the aid of a sonicator. Suitable sonicators include Vibracell VCX
750 with model CV33 probe head, Sonics and Materials Inc., Newtown,
Conn. The water phase of the emulsion is prepared by dissolving an
exendin, for example, exendin-4, and excipients such as sucrose in
water. The concentration of drug in the resulting solution can be
from about 50 mg/ml to about 100 mg/ml. For example, when the drug
is exendin-4, the concentration of drug in solution can be from
about 3.28 g to about 6.55 g per 65.5 g of water. In a particular
embodiment, 5.46 g exendin-4 and 2.18 g sucrose are dissolved in
65.5 g water for irrigation or WFI. The specified amounts listed
above represent a 4% overage to target load in order to compensate
for losses upon filter sterilization of the components. The oil
phase of the emulsion is prepared by dissolving PLGA polymer (e.g.,
97.7 g of purified 50:50 DL4A PLGA (Alkermes, Inc.)) in methylene
chloride (1539 g or 6% w/v).
[0122] The water phase is then added to the oil phase over about a
three-minute period while sonicating at 100% amplitude at ambient
temperature. The water phase containing the sucrose/exendin-4 is
charged to the coacervation reactor. Reactor is then stirred at
1400 to 1600 rpm, with additional sonication at 100% amplitude for
2 minutes, followed by a 30 second hold, and then 1 minute more of
sonication. This results in an inner emulsion droplet size of less
than 0.5 microns. It is understood that inner emulsion formation
can be achieved using any suitable means. Suitable means of
emulsion formation include, but are not limited to, sonication as
described above and homogenization.
[0123] A coacervation step is then performed by adding silicone oil
(2294 gr of Dimethicone, NF, 350 cs) over time period of less than
five minutes to the inner emulsion. This is equivalent to a ratio
of 1.5:1, silicone oil to methylene chloride. The methylene
chloride from the polymer solution partitions into the silicone oil
and begins to precipitate the polymer around the water phase
containing exendin, leading to microencapsulation. The embryonic
microspheres thus formed are soft and require hardening.
Frequently, the embryonic microspheres are permitted to stand for a
short period of time, for example, of less than 1 minute or from
about 1 minute to about 5 minutes prior to proceeding to the
microsphere hardening step.
[0124] The embryonic microspheres are then immediately transferred
into a heptane/ethanol solvent mixture. The volume of
heptane/ethanol mixture needed can be determined based on the
microsphere batch size. In the present example, about 22 kg heptane
and 2448 g ethanol in a 3.degree. C. cooled, stirred tank (350 to
450 rpm) are used. This solvent mixture hardens the microspheres by
extracting additional methylene chloride from the microspheres.
This hardening step can also be referred to as quenching. After
being quenched for 1 hour at 3.degree. C., the solvent mixture is
decanted and fresh heptane (13 Kg) is added at 3.degree. C. and
held for 1 hour to rinse off residual silicone oil, ethanol and
methylene chloride on the microsphere surface.
[0125] At the end of the rinse step, the microspheres are
transferred and collected, for example, on a 6'' diameter, 20
micron multilayered screen inside the cone shaped drying chamber
which acts as a dead-end filter. A final rinse with heptane (6 Kg
at 4.degree. C.) is performed to ensure maximum line transfer. The
microspheres are then dried with a constant purge of nitrogen gas
at a controlled rate, for example, according to the following
schedule: 18 hours at 3.degree. C.; 24 hours at 25.degree. C.; 6
hours at 35.degree. C.; and 42 hours at 38.degree. C.
[0126] After the completion of drying, the microspheres are
discharged into a teflon/stainless steel sterilized collection
vessel attached to the drying cone. The collection vessel is
sealed, removed from the drying cone and stored at -20.+-.5.degree.
C. until filling. Material remaining in the cone upon disassembly
for cleaning is taken for drug content analysis.
[0127] Non-limiting examples of specific PLG polymers suitable for
use in the general methods described above are listed below. The
listed polymers can be obtained from Lakeshore Biomaterials of
Birmingham, Ala., or Boehringer Ingelheim Pharma GmbH & Co. KG,
Germany, although other sources may be available, and can be
described as follows: Polymer 2A: Poly(lactide-co-glycolide); 50:50
lactide:glycolide ratio; 12.3 kD Mol. Wt.; IV=0.15 (dL/g). Polymer
4A: Poly(lactide-co-glycolide); 50:50 lactide:glycolide ratio; Mol.
Wt. 45-64 kD; IV=0.45-0.47 (dL/g).
[0128] It is known in the art that proteins and peptides which are
incorporated in PLG matrices can be undesirably altered (e.g.,
degraded or chemically modified) as a result of interaction with
degradation products of the PLG or impurities remaining after
preparation of the polymer. Lucke et al., Pharmaceutical Research,
19(2):175-181 (2002). As such, the PLG polymers used in the
preparation of microparticle formulations described herein can be
purified prior to preparation of the sustained release compositions
using art recognized purification methods.
EXAMPLES
[0129] In order that the invention described herein may be more
fully understood, the following examples are set forth. It should
be understood that these examples are for illustrative purposes
only and are not to be construed as limiting this invention in any
manner.
[0130] The pharmacokinetics of a long-acting release formulation of
exenatide was evaluated in a study in patients with Type 2
diabetes. The study population consisted of individuals with type 2
diabetes treated with a stable regimen of oral diabetes medications
or managed with diet modification and exercise. Patients, male or
female, had a mean age of 55.+-.10 years with a mean body mass
index (BMI) of about 34.9 kg/m.sup.2 at screening, and a mean
HbA.sub.1c of 9.3.+-.1.0% at screening. The 30 week study compared
a 10 .mu.g formulation of exenatide administered twice daily (BID)
by subcutaneous (SC) injection and a 2 mg formulation of exenatide
administered once weekly by subcutaneous (SC) injection. The study
was also conducted, inter alia, to examine the effects of such
administration on the individual's total cholesterol,
LDL-cholesterol, HDL-cholesterol, triglyceride levels, hemoglobin
A.sub.1C, fasting postprandial blood glucose, and weight.
[0131] During the 3-day lead-in period, patients self-administered
exenatide 5 .mu.g SC, BID, within 15 minutes prior to meals in the
morning and evening. The exenatide 5 .mu.g SC, BID used in the
3-day lead-in period was a clear, colorless, sterile preserved
solution for SC injection containing exenatide (exendin-4) in
sodium acetate buffer, pH 4.5, 4.3% mannitol as an iso-osmolality
modifier and 0.22% metacresol as a preservative. The strength of
exenatide injection was 0.25 mg/mL of exendin-4. This 3-day lead-in
period was designed to expose patients to exenatide prior to
administration of exenatide 10 .mu.g BID or exenatide once weekly
to determine if a patient may exhibit an acute sensitivity to
exenatide.
[0132] During the 30-week treatment period, patients received once
weekly subcutaneous injections of a 2.0 mg dose pharmaceutical
formulation containing 5% exenatide or patient received twice daily
subcutaneous injections of exenatide at 10 .mu.g per dose.
[0133] Exenatide once weekly is a sustained-release formulation of
exenatide (exendin-4) designed to provide exenatide release over a
period ranging from 7 to 91 days. Exenatide once weekly used in
this study contained, on a w/w basis, 5% exenatide, 2% sucrose, and
93% MEDISORB.RTM. 50:50 poly D,L-lactic co-glycolic acid (also
referred to as "poly(lactide-co-glycolide)"). The vial containing
the white to off-white dry powder (2.8 mg of exenatide once weekly)
was stored frozen in a freezer with a recorded temperature at
.ltoreq.-20.+-.5.degree. C. at the study site.
[0134] Patients were monitored about every four weeks for body
weight, vital signs (including blood pressure, heart rate,
respiratory rate, and temperature), urinalysis, and adverse events.
In addition, blood was drawn to assess plasma concentrations of
exendin-4 and blood lipids.
[0135] Plasma exenatide was quantified by a validated Enzyme-Linked
Immunosorbent Assay (ELISA) at Millipore (Billerica, Mass.).
Fineman et al, Diabetes Care, 26:2370-2377 (2003). Glycosylated
hemoglobin was quantitated by Quintiles Laboratories (Smyrna, Ga.)
using high-performance liquid chromatography. Davis et al,
Diabetes, 27:102-107 (1978); Cole et al, Metabolism, 27:289-301
(1978).
[0136] FIG. 1 shows the plasma concentration of exendin-4 over a
period of 30 weeks. The average plasma concentration at week 30 was
about 388 pg/mL for a 2.0 mg dose of a pharmaceutical formulation
containing 5% exendin-4. The geometric mean plasma concentration of
exendin-4 at week 30 was about 300 pg/mL for a 2.0 mg dose of a
pharmaceutical formulation containing 5% exendin-4. The range of
the geometric mean plasma concentration of exendin-4 at week 30 was
about 145 pg/mL to about 700 pg/mL for a 2.0 mg dose of a
pharmaceutical formulation containing 5% exendin-4.
[0137] For exenatide once weekly, fasting triglycerides levels
decreased by an average of 42 mg/dL from baseline to week 30 in a
group of 148 patients, as shown in FIG. 2A. For exenatide BID,
fasting triglycerides levels decreased by an average of 22 mg/dL
from baseline to week 30 in a group of 147 patients, as shown in
FIG. 2B.
[0138] For exenatide once weekly, the total cholesterol and
LDL-cholesterol decreased by an average of 11.9 mg/dL and 4.9
mg/dL, respectively from baseline to week 30, as shown in FIGS. 3A
and 3C. For exenatide BID, the total cholesterol decreased by an
average of 3.8 mg/dL from baseline to week 30, as shown in FIG. 3B,
and the LDL-cholesterol increased by an average of 1.2 mg/dL from
baseline to week 30, as shown in FIG. 3D. As shown in FIGS. 3E and
3F, HDL-cholesterol decreased by 0.9 mg/dL and 1.3 mg/dL from
baseline to week 30 for exenatide once weekly and exenatide BID,
respectively.
[0139] FIG. 4 shows the weight loss associated with exenatide once
weekly and exenatide BID. For exenatide once weekly, FIG. 4A shows
that the weight loss from baseline to week 30 was about 3.7 kg. For
exenatide BID, FIG. 4B shows that the weight loss from baseline to
week 30 was about 3.6 kg.
[0140] FIG. 5A shows that HbA.sub.1C decreased by an average of
1.9% from baseline to week 30 in patients receiving exenatide once
weekly. FIG. 5B shows that HbA.sub.1C decreased by an average of
1.5% from baseline to week 30 in patients receiving exenatide
BID.
[0141] FIG. 6A shows that fasting postprandial blood glucose
decreased by an average of 42 mg/dL from baseline to week 30 in
patient receiving exenatide once weekly, while FIG. 6B shows that
fasting postprandial blood glucose decreased by an average of 25
mg/dL from baseline to week 30 in patient receiving exenatide
BID.
[0142] All publications, patents, patent applications, and other
references cited in this application are hereby incorporated by
reference in their entirety as if each individual publication,
patent, patent application, or other reference was specifically and
individually indicated to be incorporated by reference.
[0143] The detailed description provided herein is to aid the
skilled artisan in practicing the invention. This detailed
description should not be construed to unduly limit the invention
as modifications and variations in the embodiments discussed herein
can be made by one of ordinary skill in the art without departing
from the spirit or scope of the inventive discovery.
Sequence CWU 1
1
157139PRTHeloderma horridumMOD_RES(39)..(39)AMIDATION 1His Gly Glu
Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala
Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser
Gly Ala Pro Pro Pro Ser 35239PRTHeloderma
suspectumMOD_RES(39)..(39)AMIDATION 2His Ser Asp Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro
Pro Ser 35330PRTHomo sapiensMOD_RES(30)..(30)AMIDATION 3His Ala Glu
Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly1 5 10 15Gln Ala
Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg 20 25
30431PRTHeloderma horridumMOD_RES(31)..(31)AMIDATION 4Asp Leu Ser
Lys Gln Met Glu Glu Glu Ala Val Arg Leu Phe Ile Glu1 5 10 15Trp Leu
Lys Asn Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser 20 25
30538PRTHeloderma horridum 5His Ser Asp Ala Thr Phe Thr Ala Glu Tyr
Ser Lys Leu Leu Ala Lys1 5 10 15Leu Ala Leu Gln Lys Tyr Leu Glu Ser
Ile Leu Gly Ser Ser Thr Ser 20 25 30Pro Arg Pro Pro Ser Ser
35637PRTHeloderma horridum 6His Ser Asp Ala Thr Phe Thr Ala Glu Tyr
Ser Lys Leu Leu Ala Lys1 5 10 15Leu Ala Leu Gln Lys Tyr Leu Glu Ser
Ile Leu Gly Ser Ser Thr Ser 20 25 30Pro Arg Pro Pro Ser
35735PRTHeloderma suspectumMOD_RES(35)..(35)AMIDATION 7His Ser Asp
Ala Ile Phe Thr Glu Glu Tyr Ser Lys Leu Leu Ala Lys1 5 10 15Leu Ala
Leu Gln Lys Tyr Leu Ala Ser Ile Leu Gly Ser Arg Thr Ser 20 25 30Pro
Pro Pro 35835PRTArtificial SequenceDescription of Artificial
Sequence Synthetic exendin analog polypeptide 8His Ser Asp Ala Ile
Phe Thr Gln Gln Tyr Ser Lys Leu Leu Ala Lys1 5 10 15Leu Ala Leu Gln
Lys Tyr Leu Ala Ser Ile Leu Gly Ser Arg Thr Ser 20 25 30Pro Pro Pro
35930PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 9His Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Lys Asn Gly Gly 20 25 301030PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 10His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn
Gly Gly 20 25 301128PRTArtificial SequenceDescription of Artificial
Sequence Synthetic exendin analog peptide 11His Gly Glu Gly Thr Phe
Thr Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu
Phe Ile Glu Trp Leu Lys Asn 20 251239PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 12His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro Pro Ser
351328PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 13His Gly Glu Gly Thr Phe Thr Ser
Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile
Glu Phe Leu Lys Asn 20 251428PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog peptide 14His Gly Glu
Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala
Val Arg Leu Ala Ile Glu Phe Leu Lys Asn 20 251538PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 15Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
Xaa Xaa Xaa1 5 10 15Xaa Ala Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
Gly Gly Xaa Ser 20 25 30Ser Gly Ala Xaa Xaa Xaa 351639PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 16Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
Xaa Xaa Xaa1 5 10 15Xaa Ala Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
Gly Gly Xaa Ser 20 25 30Ser Gly Ala Xaa Xaa Xaa Xaa
351738PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 17Xaa Xaa Xaa Gly Xaa Xaa Xaa
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa1 5 10 15Xaa Ala Xaa Xaa Xaa Xaa
Xaa Xaa Xaa Xaa Xaa Xaa Gly Gly Xaa Ser 20 25 30Ser Gly Ala Xaa Xaa
Xaa 351839PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 18Xaa Xaa Xaa Xaa Xaa Xaa Xaa
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa1 5 10 15Xaa Ala Xaa Xaa Xaa Xaa
Xaa Xaa Xaa Xaa Xaa Xaa Gly Gly Xaa Ser 20 25 30Ser Gly Ala Xaa Xaa
Xaa Ser 351941PRTArtificial SequenceDescription of Artificial
Sequence Synthetic exendin analog polypeptide 19Xaa Xaa Xaa Gly Xaa
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa1 5 10 15Xaa Ala Xaa Xaa
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly 20 25 30Gly Xaa Ser
Ser Gly Ala Xaa Xaa Xaa 35 402042PRTArtificial SequenceDescription
of Artificial Sequence Synthetic exendin analog polypeptide 20Xaa
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa1 5 10
15Xaa Ala Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly
20 25 30Gly Xaa Ser Ser Gly Ala Xaa Xaa Xaa Xaa 35
402139PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 21Xaa Xaa Xaa Gly Thr Xaa Xaa
Xaa Xaa Xaa Ser Lys Gln Xaa Glu Glu1 5 10 15Glu Ala Val Arg Leu Xaa
Xaa Xaa Xaa Leu Lys Asn Gly Gly Xaa Ser 20 25 30Ser Gly Ala Xaa Xaa
Xaa Xaa 352241PRTArtificial SequenceDescription of Artificial
Sequence Synthetic exendin analog polypeptide 22Xaa Xaa Xaa Gly Thr
Xaa Xaa Xaa Xaa Xaa Ser Lys Gln Xaa Glu Glu1 5 10 15Glu Ala Val Arg
Leu Xaa Xaa Xaa Xaa Leu Xaa Xaa Xaa Xaa Gly Gly 20 25 30Xaa Ser Ser
Gly Ala Xaa Xaa Xaa Xaa 35 402330PRTArtificial SequenceDescription
of Artificial Sequence Synthetic exendin analog polypeptide 23His
Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10
15Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly 20 25
302428PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 24His Gly Glu Gly Thr Phe Thr Ser
Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile
Glu Trp Leu Lys Asn 20 252528PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog peptide 25His Gly Glu
Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala
Val Arg Leu Phe Ile Glu Phe Leu Lys Asn 20 252628PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
peptide 26His Ala Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu
Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn 20
252728PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 27His Gly Glu Gly Ala Phe Thr Ser
Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile
Glu Phe Leu Lys Asn 20 252828PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog peptide 28His Gly Glu
Gly Thr Ala Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala
Val Arg Leu Phe Ile Glu Phe Leu Lys Asn 20 252928PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
peptide 29His Gly Glu Gly Thr Phe Thr Ala Asp Leu Ser Lys Gln Leu
Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn 20
253028PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 30His Gly Glu Gly Thr Phe Thr Ser
Asp Ala Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile
Glu Phe Leu Lys Asn 20 253128PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog peptide 31His Gly Glu
Gly Thr Phe Thr Ser Asp Leu Ala Lys Gln Leu Glu Glu1 5 10 15Glu Ala
Val Arg Leu Phe Ile Glu Phe Leu Lys Asn 20 253228PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
peptide 32His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Ala Gln Leu
Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn 20
253328PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 33His Gly Glu Gly Thr Phe Thr Ser
Asp Leu Ser Lys Ala Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile
Glu Phe Leu Lys Asn 20 253428PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog peptide 34His Gly Glu
Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Ala Glu Glu1 5 10 15Glu Ala
Val Arg Leu Phe Ile Glu Phe Leu Lys Asn 20 253528PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
peptide 35His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu
Ala Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn 20
253628PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 36His Gly Glu Gly Thr Phe Thr Ser
Asp Leu Ser Lys Gln Leu Glu Ala1 5 10 15Glu Ala Val Arg Leu Phe Ile
Glu Phe Leu Lys Asn 20 253728PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog peptide 37His Gly Glu
Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Ala Ala
Val Arg Leu Phe Ile Glu Phe Leu Lys Asn 20 253828PRTArtificial
SequenceDescription of Artificial Sequence Synthetic novel exendin
agonist compound peptide 38His Gly Glu Gly Thr Phe Thr Ser Asp Leu
Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala Ala Arg Leu Phe Ile Glu Phe
Leu Lys Asn 20 253928PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog peptide 39His Gly Glu
Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala
Val Ala Leu Phe Ile Glu Phe Leu Lys Asn 20 254028PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
peptide 40His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu
Glu Glu1 5 10 15Glu Ala Val Arg Ala Phe Ile Glu Phe Leu Lys Asn 20
254128PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 41His Gly Glu Gly Thr Phe Thr Ser
Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile
Ala Phe Leu Lys Asn 20 254228PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog peptide 42His Gly Glu
Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala
Val Arg Leu Phe Ile Glu Ala Leu Lys Asn 20 254328PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
peptide 43His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu
Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Ala Lys Asn 20
254428PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 44His Gly Glu Gly Thr Phe Thr Ser
Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile
Glu Phe Leu Ala Asn 20 254528PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog peptide 45His Gly Glu
Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala
Val Arg Leu Phe Ile Glu Phe Leu Lys Ala 20 254638PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 46His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro Pro 354738PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 47His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro Pro 354837PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 48His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro 354937PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 49His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro 355036PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 50His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro 355136PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 51His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro 355235PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 52His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala 355335PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 53His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala 355434PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 54His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly5534PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 55His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Leu Glu Glu1 5
10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn Gly Gly Pro
Ser 20 25 30Ser Gly5633PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog polypeptide 56His Gly
Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu
Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25
30Ser5733PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 57His Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Phe Leu Lys Asn Gly Gly Pro Ser 20 25
30Ser5832PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 58His Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 305932PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 59His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn
Gly Gly Pro Ser 20 25 306031PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog polypeptide 60His Gly
Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu
Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro 20 25
306131PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 61His Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Phe Leu Lys Asn Gly Gly Pro 20 25 306230PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 62His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn
Gly Gly 20 25 306329PRTArtificial SequenceDescription of Artificial
Sequence Synthetic exendin analog peptide 63His Gly Glu Gly Thr Phe
Thr Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu
Phe Ile Glu Trp Leu Lys Asn Gly 20 256429PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
peptide 64His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu
Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn Gly
20 256538PRTArtificial SequenceDescription of Artificial Sequence
Synthetic polypeptide 65His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser
Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Trp Leu
Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro Pro
356638PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 66His Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro
Pro 356737PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 67His Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Lys Asn Gly Gly Ala Ser 20 25 30Ser Gly Ala Pro Pro
356837PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 68His Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Lys Asn Gly Gly Ala Ser 20 25 30Ser Gly Ala Ala Ala
356937PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 69His Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro
357036PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 70His Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro
357135PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 71Arg Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly Ala
357230PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 72His Gly Asp Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Lys Asn Gly Gly 20 25 307328PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
peptide 73His Gly Glu Gly Thr Ala Thr Ser Asp Leu Ser Lys Gln Leu
Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn 20
257428PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 74His Gly Glu Gly Thr Phe Ser Ser
Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile
Glu Trp Leu Lys Asn 20 257528PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog peptide 75His Gly Glu
Gly Thr Phe Ser Thr Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala
Val Arg Leu Phe Ile Glu Trp Leu Lys Asn 20 257628PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
peptide 76His Gly Glu Gly Thr Phe Thr Ser Glu Leu Ser Lys Gln Met
Ala Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn 20
257728PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 77His Gly Glu Gly Thr Phe Thr Ser
Asp Gly Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile
Glu Phe Leu Lys Asn 20 257828PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog peptide 78His Gly Glu
Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala
Val Arg Leu Ala Ile Glu Phe Leu Lys Asn 20 257928PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
peptide 79His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met
Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Gly Glu Trp Leu Lys Asn 20
258028PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 80His Gly Glu Gly Thr Phe Thr Ser
Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile
Asp Phe Leu Lys Asn 20 258133PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog polypeptide 81His Gly
Glu Gly Thr Phe Thr Ser Asp Ala Ser Lys Gln Leu Glu Glu1 5 10 15Glu
Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn Gly Gly Pro Ser 20 25
30Ser8229PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 82His Gly Glu Gly Thr Phe Thr Ser
Asp Ala Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile
Glu Trp Leu Lys Asn Gly 20 258337PRTArtificial SequenceDescription
of Artificial Sequence Synthetic exendin analog polypeptide 83His
Gly Glu Gly Thr Phe Thr Ser Asp Ala Ser Lys Gln Met Glu Glu1 5 10
15Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser
20 25 30Ser Gly Ala Pro Pro 358427PRTArtificial SequenceDescription
of Artificial Sequence Synthetic exendin analog peptide 84Gly Glu
Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu Glu1 5 10 15Ala
Val Arg Leu Phe Ile Glu Trp Leu Lys Asn 20 258527PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
peptide 85Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu
Glu Glu1 5 10 15Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn 20
258629PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 86Gly Glu Gly Thr Phe Thr Ser Asp
Leu Ser Lys Gln Met Glu Glu Glu1 5 10 15Ala Val Arg Leu Phe Ile Glu
Trp Leu Lys Asn Gly Gly 20 258729PRTArtificial SequenceDescription
of Artificial Sequence Synthetic exendin analog peptide 87Gly Glu
Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu Glu1 5 10 15Ala
Val Arg Leu Phe Ile Glu Phe Leu Lys Asn Gly Gly 20
258827PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 88Gly Glu Gly Thr Phe Thr Ser Asp
Leu Ser Lys Gln Met Glu Glu Glu1 5 10 15Ala Val Arg Leu Phe Ile Glu
Trp Leu Asn Lys 20 258927PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog peptide 89Gly Glu Gly
Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu Glu1 5 10 15Ala Val
Arg Leu Phe Ile Glu Phe Leu Asn Lys 20 259029PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
peptide 90Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu
Glu Glu1 5 10 15Ala Val Arg Leu Phe Ile Glu Trp Leu Asn Lys Gly Gly
20 259129PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 91Gly Glu Gly Thr Phe Thr Ser Asp
Leu Ser Lys Gln Leu Glu Glu Glu1 5 10 15Ala Val Arg Leu Phe Ile Glu
Phe Leu Asn Lys Gly Gly 20 259227PRTArtificial SequenceDescription
of Artificial Sequence Synthetic exendin analog peptide 92Gly Glu
Gly Thr Phe Thr Ser Ala Leu Ser Lys Gln Met Glu Glu Glu1 5 10 15Ala
Val Arg Leu Phe Ile Glu Trp Leu Lys Asn 20 259327PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
peptide 93Gly Glu Gly Thr Phe Thr Ser Ala Leu Ser Lys Gln Leu Glu
Glu Glu1 5 10 15Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn 20
259429PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 94Gly Glu Gly Thr Phe Thr Ser Ala
Leu Ser Lys Gln Met Glu Glu Glu1 5 10 15Ala Val Arg Leu Phe Ile Glu
Trp Leu Lys Asn Gly Gly 20 259529PRTArtificial SequenceDescription
of Artificial Sequence Synthetic exendin analog peptide 95Gly Glu
Gly Thr Phe Thr Ser Ala Leu Ser Lys Gln Leu Glu Glu Glu1 5 10 15Ala
Val Arg Leu Phe Ile Glu Phe Leu Lys Asn Gly Gly 20
259627PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 96Gly Glu Gly Thr Phe Thr Ser Ala
Leu Ser Lys Gln Met Glu Glu Glu1 5 10 15Ala Val Arg Leu Phe Ile Glu
Trp Leu Asn Lys 20 259727PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog peptide 97Gly Glu Gly
Thr Phe Thr Ser Ala Leu Ser Lys Gln Leu Glu Glu Glu1 5 10 15Ala Val
Arg Leu Phe Ile Glu Phe Leu Asn Lys 20 259829PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
peptide 98Gly Glu Gly Thr Phe Thr Ser Ala Leu Ser Lys Gln Met Glu
Glu Glu1 5 10 15Ala Val Arg Leu Phe Ile Glu Trp Leu Asn Lys Gly Gly
20 259929PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog peptide 99Gly Glu Gly Thr Phe Thr Ser Ala
Leu Ser Lys Gln Leu Glu Glu Glu1 5 10 15Ala Val Arg Leu Phe Ile Glu
Phe Leu Asn Lys Gly Gly 20 2510028PRTArtificial SequenceDescription
of Artificial Sequence Synthetic exendin analog peptide 100Ala Gly
Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu
Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn 20 2510128PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
peptide 101Ala Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu
Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn 20
2510230PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 102Ala Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Lys Asn Gly Gly 20 25 3010330PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 103Ala Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn
Gly Gly 20 25 3010428PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog peptide 104Ala Gly Glu
Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala
Val Arg Leu Phe Ile Glu Trp Leu Asn Lys 20 2510528PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
peptide 105Ala Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu
Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Asn Lys 20
2510630PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 106Ala Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Asn Lys Gly Gly 20 25 3010730PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 107Ala Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Asn Lys
Gly Gly 20 25 3010839PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog polypeptide 108His Gly
Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu
Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn Gly Gly Pro Ser 20 25
30Ser Gly Ala Pro Pro Pro Ser 3510939PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 109His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro Pro Ser
3511039PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 110His Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala
Val Arg Leu Phe Ile Glu Phe Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser
Gly Ala Pro Pro Pro Ser 3511139PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog polypeptide 111Tyr Gly
Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu
Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25
30Ser Gly Ala Pro Pro Pro Ser 3511239PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 112His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro Pro Tyr
3511339PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 113His Gly Asp Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro
Pro Ser 3511439PRTArtificial SequenceDescription of Artificial
Sequence Synthetic exendin analog polypeptide 114His Gly Glu Gly
Thr Ala Thr Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val
Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly
Ala Pro Pro Pro Ser 3511539PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog polypeptide 115His Gly
Glu Gly Thr Phe Ser Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu
Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25
30Ser Gly Ala Pro Pro Pro Ser 3511639PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 116His Gly Glu Gly Thr Phe Ser Thr Asp Leu Ser Lys Gln
Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro Pro Ser
3511739PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 117His Gly Glu Gly Thr Phe Thr
Thr Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro
Pro Ser 3511839PRTArtificial SequenceDescription of Artificial
Sequence Synthetic exendin analog polypeptide 118His Gly Glu Gly
Thr Phe Thr Ser Glu Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val
Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly
Ala Pro Pro Pro Ser 3511939PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog polypeptide 119His Gly
Glu Gly Thr Phe Thr Ser Asp Gly Ser Lys Gln Met Glu Glu1 5 10 15Glu
Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25
30Ser Gly Ala Pro Pro Pro Ser 3512039PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 120His Gly Glu Gly Thr Phe Thr Ser Asp Gly Ser Lys Gln
Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro Pro Ser
3512139PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 121His Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Gly Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro
Pro Ser 3512239PRTArtificial SequenceDescription of Artificial
Sequence Synthetic exendin analog polypeptide 122His Gly Glu Gly
Thr Phe Thr Ser Asp Leu Ser Lys Gln Gly Glu Glu1 5 10 15Glu Ala Val
Arg Leu Phe Ile Glu Phe Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly
Ala Pro Pro Pro Ser 3512339PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog polypeptide 123His Gly
Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu
Ala Val Arg Leu Ala Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25
30Ser Gly Ala Pro Pro Pro Ser 3512439PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 124His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Val Glu Trp Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro Pro Ser
3512539PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 125His Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Val Glu Phe Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro
Pro Ser 3512639PRTArtificial SequenceDescription of Artificial
Sequence Synthetic exendin analog polypeptide 126His Gly Glu Gly
Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val
Arg Leu Phe Gly Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly
Ala Pro Pro Pro Ser 3512739PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog polypeptide 127His Gly
Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu
Ala Val Arg Leu Phe Gly Glu Phe Leu Lys Asn Gly Gly Pro Ser 20 25
30Ser Gly Ala Pro Pro Pro Ser 3512839PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 128His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Asp Trp Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro Pro Ser
3512939PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 129His Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Phe Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro
Pro Ser 3513039PRTArtificial SequenceDescription of Artificial
Sequence Synthetic exendin analog polypeptide 130His Gly Glu Gly
Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val
Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly
Ala Pro Pro Pro Ser 3513139PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog polypeptide 131His Gly
Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu
Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25
30Ser Gly Ala Pro Pro Pro Ser 3513239PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 132His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro Pro Ser
3513339PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 133His Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro
Pro Ser 3513439PRTArtificial SequenceDescription of Artificial
Sequence Synthetic exendin analog polypeptide 134His Gly Glu Gly
Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala Val
Arg Leu Phe Ile Glu Phe Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly
Ala Pro Pro Pro Ser 3513539PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog polypeptide 135His Gly
Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu
Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn Gly Gly Pro Ser 20 25
30Ser Gly Ala Pro Pro Pro Ser 3513639PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 136His Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn
Gly Gly Ala Ser 20 25 30Ser Gly Ala Ala Ala Ala Ser
3513739PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 137His Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25 30Ser Gly Ala Ala Ala
Ala Ser 3513839PRTArtificial SequenceDescription of Artificial
Sequence Synthetic exendin analog polypeptide 138His Gly Glu Gly
Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala Val
Arg Leu Phe Ile Glu Phe Leu Lys Asn Gly Gly Ala Ser 20 25 30Ser Gly
Ala Ala Ala Ala Ser 3513939PRTArtificial SequenceDescription of
Artificial Sequence Synthetic exendin analog polypeptide 139Xaa Gly
Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu
Ala Val Arg Leu Phe Ile Glu Trp Leu Lys Asn Gly Gly Pro Ser 20 25
30Ser Gly Ala Pro Pro Pro Ser 3514039PRTArtificial
SequenceDescription of Artificial Sequence Synthetic exendin analog
polypeptide 140Xaa Gly Glu Gly Thr Phe Thr Ser Asp Leu Ser Lys Gln
Leu Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe Ile Glu Phe Leu Lys Asn
Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro Pro Ser
3514139PRTArtificial SequenceDescription of Artificial Sequence
Synthetic exendin analog polypeptide 141Xaa Gly Glu Gly Thr Phe Thr
Ser Asp Leu Ser Lys Gln Met Glu Glu1 5 10 15Glu Ala Val Arg Leu Phe
Ile Glu Trp Leu Asn Lys Gly Gly Pro Ser 20 25 30Ser Gly Ala Pro Pro
Pro Ser 3514239PRTArtificial SequenceDescription of Artificial
Sequence Synthetic exendin analog polypeptide 142Xaa Gly Glu Gly
Thr Phe Thr Ser Asp Leu Ser Lys Gln Leu Glu Glu1 5 10 15Glu Ala Val
Arg Leu Phe Ile Glu Phe Leu Asn Lys Gly Gly Pro Ser 20 25 30Ser Gly
Ala Pro Pro Pro Ser 351435PRTArtificial SequenceDescription of
Artificial Sequence Synthetic peptide 143Gly Gly Xaa Ser Ser1
51446PRTArtificial SequenceDescription of Artificial Sequence
Synthetic peptide 144Gly Gly Xaa Ser Ser Gly1 51457PRTArtificial
SequenceDescription of Artificial Sequence Synthetic peptide 145Gly
Gly Xaa Ser Ser Gly Ala1 51468PRTArtificial SequenceDescription of
Artificial Sequence Synthetic peptide 146Gly Gly Xaa Ser Ser Gly
Ala Xaa1 51479PRTArtificial SequenceDescription of Artificial
Sequence Synthetic peptide 147Gly Gly Xaa Ser Ser Gly Ala Xaa Xaa1
514810PRTArtificial SequenceDescription of Artificial Sequence
Synthetic peptide 148Gly Gly Xaa Ser Ser Gly Ala Xaa Xaa Xaa1 5
101495PRTArtificial SequenceDescription of Artificial Sequence
Synthetic peptide 149Gly Gly Xaa Ser Ser1 51506PRTArtificial
SequenceDescription of Artificial Sequence Synthetic peptide 150Gly
Gly Xaa Ser Ser Gly1 51517PRTArtificial SequenceDescription of
Artificial Sequence Synthetic peptide 151Gly Gly Xaa Ser Ser Gly
Ala1 51528PRTArtificial SequenceDescription of Artificial Sequence
Synthetic peptide 152Gly Gly Xaa Ser Ser Gly Ala Xaa1
51539PRTArtificial SequenceDescription of Artificial Sequence
Synthetic peptide 153Gly Gly Xaa Ser Ser Gly Ala Xaa Xaa1
515410PRTArtificial SequenceDescription of Artificial Sequence
Synthetic peptide 154Gly Gly Xaa Ser Ser Gly Ala Xaa Xaa Xaa1 5
1015511PRTArtificial SequenceDescription of Artificial Sequence
Synthetic peptide 155Gly Gly Xaa Ser Ser Gly Ala Xaa Xaa Xaa Xaa1 5
1015611PRTArtificial SequenceDescription of Artificial Sequence
Synthetic peptide 156Gly Gly Xaa Ser Ser Gly Ala Xaa Xaa Xaa Xaa1 5
1015711PRTArtificial SequenceDescription of Artificial Sequence
Synthetic peptide 157Gly Gly Xaa Ser Ser Gly Ala Xaa Xaa Xaa Ser1 5
10
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