U.S. patent application number 15/997965 was filed with the patent office on 2019-05-09 for method of restoring the incretin effect.
The applicant listed for this patent is Neurendo Pharma LLC. Invention is credited to Anton H. Clemens.
Application Number | 20190134023 15/997965 |
Document ID | / |
Family ID | 39325299 |
Filed Date | 2019-05-09 |
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United States Patent
Application |
20190134023 |
Kind Code |
A1 |
Clemens; Anton H. |
May 9, 2019 |
Method of Restoring the Incretin Effect
Abstract
The present invention relates to methods of treating metabolic
syndrome, Type 2 diabetes mellitus, atherogenic dyslipidemia and/or
obesity. The present invention a so relates to methods of restoring
the incretin effect, to restoring physiologic control of glucagon
levels, to restoring first-phase insulin secretion, and to
restoring the physiologic glucose-dependent insulin secretion. The
methods of the present invention comprise administration of a
selective .kappa.-receptor antagonist, such as guanidinylated
naltrindole (GNTI), or pharmaceutically acceptable derivatives
thereof to a subject in need thereof.
Inventors: |
Clemens; Anton H.; (Madison,
WI) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Neurendo Pharma LLC |
Madison |
WI |
US |
|
|
Family ID: |
39325299 |
Appl. No.: |
15/997965 |
Filed: |
June 5, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14479828 |
Sep 8, 2014 |
9987268 |
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15997965 |
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13859940 |
Apr 10, 2013 |
8829018 |
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14479828 |
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12906735 |
Oct 18, 2010 |
8445508 |
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13859940 |
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11876279 |
Oct 22, 2007 |
7893080 |
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12906735 |
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60862227 |
Oct 20, 2006 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 3/10 20180101; A61K
38/45 20130101; A61P 3/00 20180101; A61K 31/4748 20130101; A61P
3/04 20180101; A61K 31/485 20130101; A61K 31/437 20130101; Y10S
514/866 20130101; A61K 45/06 20130101; A61P 3/08 20180101 |
International
Class: |
A61K 31/485 20060101
A61K031/485; A61K 45/06 20060101 A61K045/06; A61K 38/45 20060101
A61K038/45; A61K 31/4748 20060101 A61K031/4748; A61K 31/437
20060101 A61K031/437 |
Claims
1. A method of treating type 2 diabetes mellitus comprising
administering to a subject a therapeutically effective amount of a
selective .kappa.-receptor antagonist, or a pharmaceutically
acceptable derivative thereof.
2. The method of claim 1, wherein the selective .kappa.-receptor
antagonist is GNTI.
3. The method of claim 1, wherein the selective .kappa.-receptor
antagonist is administered weekly or daily.
4. The method of claim 3, wherein the selective .kappa.-receptor
antagonist is administered weekly in an amount from about 30 ng to
about 300 ng per kg of body weight weekly.
5. The method of claim 3, wherein the selective .kappa.-receptor
antagonist is administered daily in an amount from about 8 ng to
about 80 ng per kg of body weight daily.
6. The method of claim 1, wherein the selective .kappa.-receptor
antagonist is administered sublingually, orally, enterally,
parenterally, topically, systemically or injected intravascularly,
subcutaneously, peritoneally.
7. The method of claim 1, further comprising co-administration of
an effective amount of an insulinogenic agent.
8. The method of claim 7, wherein the insulinogenic agent is an
extended release composition.
9. The method of claim 1, wherein a .mu.-agonist is not
co-administered.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional patent application of U.S.
patent application Ser. No. 14/479,828 filed Sep. 8, 2014, which is
a continuation of U.S. patent application Ser. No. 13/859,940 filed
Apr. 10, 2013 and issued as U.S. Pat. No. 8,829,018, which is a
continuation patent application of U.S. patent application Ser. No.
12/906,735 filed Oct. 18, 2010 and issued as U.S. Pat. No.
8,445,508 on May 21, 2013, which is a divisional patent application
of application Ser. No. 11/876,279 filed Oct. 22, 2007 and issued
as U.S. Pat. No. 7,893,080 on Feb. 22, 2011, which is a
non-provisional application of U.S. Provisional Application No.
60/862,227 filed Oct. 22, 2006. The Ser. No. 11/876,279 application
claims priority to U.S. Provisional Application No. 60/862,227
filed Oct. 22, 2006. The above-referenced applications are
incorporated herein in their entireties.
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH
[0002] Not applicable.
INTRODUCTION
[0003] Insulin secretion is stimulated to greater extent by oral
intake of glucose, than by intravenous intake of glucose. This
effect, which is called the incretin effect, is estimated to be
responsible for more than half of the insulin response to glucose.
The incretin effect is caused mainly by the two intestinal
insulin-stimulating hormones, glucagon-like peptide-1 (GLP)-1) and
glucose-dependent insulinotropic polypeptide (GIP). In patients
with Type 2 diabetes mellitus, and other components of the
metabolic syndrome, such as impaired glucose tolerance, atherogenic
dyslipidemia, overweight and obesity, the incretin effect is either
greatly impaired or absent.
[0004] The rapid and sizable increase in insulin release initiated
by the incretin effect begins within two minutes of nutrient
ingestion and continues for up to 15 minutes. This post-meal
increase in insulin is referred to as first phase insulin
secretion. A second phase of insulin secretion follows and is
sustained until normal blood glucose levels are restored. Clinical
observations reveal that patients with metabolic syndrome disorders
such as Type 2 diabetes mellitus, impaired glucose tolerance, and
obesity are characterized by progressive reductions in the
magnitude of first-phase, insulin secretion, insulin resistance,
and beta-cell dysfunction, creating a new pathogenic platform
shared by all components of the metabolic syndrome. Beta-cell
dysfunction is, in turn, characterized by its two contributing
components: (1) the progressive impairment of insulin production,
and (2) the progressive impairment of physiologic control of
insulin release. It follows, that the metabolic disorders should be
correctable by the treatment, or restoration, of the failing
components of the underlying pathogenic platform of insulin
resistance and beta-cell dysfunction.
[0005] Glucose intolerance includes a continuous range of
impairments in carbohydrate metabolism. Type 2 diabetes mellitus,
impaired glucose tolerance, and impaired fasting glucose, among
other conditions associated with glucose intolerance, have been
implicated as risk factors contributing to heart disease, stroke,
overweight, obesity, hypertension, and atherogentic
dyslipidemia.
[0006] The pathogenesis of obesity is associated with other
components of the metabolic syndrome, like atherogenic
dyslipidemia, and glucose intolerance, the magnitude of which may
progress over time. Threshold levels for pharmacological treatment
have, therefore, been revised downward on several occasions in
order to intervene at an earlier stage in the epidemic expansion of
the affected population, and the corresponding health care
expenditures. Non-insulin therapies are available to reduce
endogenous gluconeogenesis or improve peripheral insulin
sensitivity, e.g., metformin, sulfonylureas or thiazolidinediones
(TZD). However, these therapies fail to restore first-phase insulin
release or the incretin effect. Importantly, enhanced early insulin
release is associated with improved overall glucose tolerance.
There is, currently, no treatment available to restore or
reactivate the natural physiology of the native incretin
system.
SUMMARY
[0007] The present invention relates to methods of treating
metabolic syndrome, Type 2 diabetes atherogenic dyslipidemia and/or
obesity. The present invention also relates to methods of restoring
the incretin effect, to restoring physiologic control of glucagon
levels, to restoring first-phase insulin secretion, and to
restoring the physiologic glucose-dependent secretion.
[0008] The methods of the present invention comprise administration
of a selective .kappa.-receptor antagonist such as guanidinylated
naltrindole (GNTI), or a pharmaceutically acceptable derivative
thereof, to a subject in need thereof. The selective
.kappa.-receptor antagonist may be administered daily, weekly or at
any suitable time interval. Administration may be sublingually,
orally, enterally, parenterally, topically or systemically. The
selective .kappa.-receptor antagonist may be co-administered with
an insulinotropic agent. The insulinotropic agent may be an
extended release formulation.
BRIEF DESCRIPTION OF FIGURES
[0009] FIG. 1 illustrates the blood glucose readings of a male
subject treated with GNTI over an extended period of time as
described in Example 1.
[0010] FIG. 2 illustrates the blood glucose readings of a male
subject treated with GNTI over a modal week as described in Example
1.
[0011] FIG. 3 illustrates the blood glucose readings of a male
subject treated with GNTI over a modal day as described in Example
1.
DETAILED DESCRIPTION
[0012] It has surprisingly been discovered that the administration
of a selective .kappa.-receptor antagonist, such as GNTI, is useful
in the treatment of metabolic syndrome by targeting the incretin
effect. The term "selective .kappa.-receptor antagonist" means a
.kappa.-receptor antagonist characterized by its .mu./.kappa.
antagonism K.sub.c selectivity ratio, as defined in J. Med. Chem
43, 2759-2769 (2000), the ratio required for this method being
<35. Suitably, the selective .kappa.-receptor antagonist acts
peripherally. That is, it is substantially free of CNS
activity.
[0013] The term "metabolic syndrome" may include, but is not
limited to, atherogenic dyslipidemia, pre-diabetes,
overweight/obesity, Type 2 diabetes mellitus and essential
hypertension. The pathogenesis of obesity is associated with other
components of the metabolic syndrome, e.g., atherogenic
dyslipidemia, and glucose intolerance, the magnitude of which may
progress, from its initial stages characterized by impaired fasting
glucose, followed by impaired glucose tolerance and culminating in
Type 2 diabetes mellitus. Administration of a selective
.kappa.-receptor antagonist, or a pharmaceutically acceptable
derivative thereof has been found to restore the incretin effect,
restore physiological control of glucagon levels in response to
ingested nutrition, restore first-phase insulin secretion, restore
glucose-dependent insulin secretion, reduce weight gain and/or
lower weight in a subject without co-administration of a
.mu.-agonist.
[0014] As will be appreciated, the methods described herein may be
useful in both research and clinical settings, suitably wherein
treatment of certain disease states are implicated, including, but
not limited to, impaired glucose tolerance, Type 2 diabetes
mellitus, diminished or absent first-phase insulin secretion, and
obesity.
[0015] The administration of a selective .kappa.-receptor
antagonist to a subject in need thereof may treat metabolic
syndrome. Administration of a selective .kappa.-receptor antagonist
may treat metabolic syndrome by restoring the incretin effect, by
restoring physiologic control of glucagon levels, by restoring the
physiologic glucose dependent insulin secretion, and/or by
restoring first-phase insulin secretion. Suitably, the
administration of a selective .kappa.-receptor antagonist may also
treat overweight, atherogenic dyslipidemia obesity or Type 2
diabetes mellitus by restoring the incretin effect, by restoring
physiologic control of glucagon levels, by restoring the
physiologic glucose dependent insulin secretion, and/or by
restoring first-phase insulin secretion.
[0016] The administration of a selective .kappa.-receptor
antagonist to a subject in need thereof restores the incretin
effect. In a subject having a normal response to oral nutrient
administration, the release of the insulinotropic hormones, GIP and
GLP-1, results in an increase in insulin secretion. This is called
the "incretin effect". As used herein, to "restore," for example,
with respect to the incretin effect, suitably includes enhancing,
potentiating, increasing, reestablishing, re-activating, or
improving the physiological state. For example, a subject having
Type 2 diabetes mellitus may exhibit diminished or even zero
incretin effect, i.e., diminished or no activity of GIP or GLP-1,
or diminished or no increase in insulin secretion upon nutrient
administration. Consequently, to "restore" the incretin effect
suitably increases, though does not necessarily normalize, GIP or
GLP-1 activity or insulin secretion upon nutrient administration in
a subject. The subject is suitably a mammal, such as a human, dog,
cat, primate, etc.
[0017] The administration of a selective .kappa.-receptor
antagonist may restore physiologic control of glucagon levels in a
subject in need thereof. As used herein, to "restore," for example
with respect to physiologic control of glucagon levels, suitably
includes, decreasing, lowering, regulating, reestablishing, or
improving the physiologic state. In a subject having a `normal`
physiologic response to nutrient administration, physiologic
control of glucagon primarily responds to blood glucose levels,
i.e., as blood glucose levels decline, glucagon is released from
the a cells of the islets of Langerhans in the pancreas, and act on
the liver to induce gluconeogenesis, i.e., endogenous glucose
production, and/or glycogenolysis. Conversely, glucagon release
decreases in response to increasing blood glucose levels.
Additionally, glucagon levels decrease in response to release of
insulin by pancreatic .beta.-cells. Consequently, in a subject
having abnormal insulin production or release in response to
increasing blood glucose levels, glucagon release may remain
abnormally high and result hyperglucagonemia which further
exacerbates conditions such as Type 2 diabetes mellitus and
impaired glucose tolerance.
[0018] Normal insulin secretion from the pancreatic .beta. cells is
biphasic. The initial release of insulin that acts on the
pancreatic .alpha. cells to decrease glucagon is referred to as the
first-phase of insulin secretion. First-phase insulin secretion is
characterized by a rapid and sizable increase in insulin, beginning
within two minutes of nutrient ingestion, and continuing for 10-15
minutes. For example, in Examples 3 and 4, first-phase insulin
secretion can be seen at the 5-minute time point coupled with a
corresponding drop in glucagon levels. Two minutes later, blood
glucose levels show an initial decrease. The second phase of
insulin secretion follows and insulin secretion peaks approximately
1-2 hours following nutrient ingestion. Insulin secretion continues
until normal blood glucose levels are restored. Often, in subjects
having impaired glucose tolerance, first-phase insulin secretion is
reduced and it is believed that the reduction in first-phase
insulin secretion may be a preliminary sign in the progression of
Type 2 diabetes mellitus.
[0019] The administration of a selective .kappa.-receptor
antagonist restores first-phase insulin secretion in a subject in
need thereof. The administration of a selective .kappa.-receptor
antagonist may restore the physiologic glucose dependent insulin
secretion in a subject in need thereof, and a selective
.kappa.-receptor antagonist may restore the physiologic control of
glucagon release.
[0020] The methods described herein restore the incretin effect,
first-phase insulin secretion and/or physiologic insulin secretion
through administration a selective .kappa.-receptor antagonist,
Suitably, the present invention may also provide a method of
treating Type 2 diabetes mellitus, atherogenic dyslipidemia,
obesity/overweight or metabolic syndrome through administration of
a selective .kappa.-receptor antagonist. The administration of an
effective amount of a selective .kappa.-receptor antagonist may
reduce weight gain or lower weight in a subject in need
thereof.
[0021] Suitably, an insulinogenic agent may be used in combination
with a selective .kappa.-receptor antagonist. An "insulinogenic
agent" stimulates, participates in the stimulation of, or
potentiates the biosynthesis of insulin by the pancreatic
.beta.-cells. Examples of insulinogenic agents include
sulfonylureas, repaglinide, nateglinide, mitiginide and BTS-67-582.
Suitably, the insulinogenic agent is provided in an extended
release composition, i.e., the insulinogenic agent is formulated
such that it is released over a period of time. An extended release
insulinogenic agent acts to potentiate the synthesis of
insulin.
[0022] Pharmacologically equivalent derivatives of a selective
.kappa.-receptor antagonist include any pharmaceutically acceptable
salts, hydrates, esters, ethers, amides, or any other derivative
which is not biologically or otherwise undesirable and induces the
desired pharmacological and/or physiological effect.
[0023] The selective .kappa.-receptor antagonist and the
insulinogenic agent (together referred to as "active agent") are
suitably administered in a pharmaceutical composition, which
include the active agent(s) and one or more pharmaceutically
acceptable excipients such as stabilizers, anti-oxidants, binders,
coloring agents, emulsifiers. The pharmaceutical composition may be
administered as a solution, an emulsion, a suspension, a
dispersion, a transdermal patch, a pill, a tablet or a capsule. One
of ordinary skill in the art would be able to formulate the
pharmaceutical composition using the appropriate solid, liquid or
gel carriers. The selective .kappa.-receptor antagonist and the
insulinogenic agent may be formulated separately or together.
[0024] Various methods for administration of the active agent(s)
may be employed. For example, the active agent(s) may be given
sublingually, orally, enterally, parenterally, topically,
systemically or may be injected intravascularly, subcutaneously,
peritoneally, and so forth. The active agent(s) may be administered
weekly, semi-weekly, daily, or multiple times a day, such as twice
a day or three times a day. The selective .kappa.-receptor
antagonist and the insulinogenic agent may be administered
concurrently. Alternatively, the selective .kappa.-receptor
antagonist may be administered before or after administration of
the insulinogenic agent.
[0025] The dosage of a selective .kappa.-receptor antagonist will
vary widely, depending upon the frequency of administration, the
manner of administration, and the clearance of a selective
.kappa.-receptor antagonist from the subject. It will be
appreciated that the specific dosage administered in any given case
will be adjusted in accordance with the condition of the subject
and other relevant medical factors that may modify the activity of
the selective .kappa.-receptor antagonist. For example, the
specific dose for a particular patient depends on age, body weight,
general state of health, diet, the timing and mode of
administration, the rate of excretion and medicaments used in
combination. For example, a suitable weekly dose of a selective
.kappa.-receptor antagonist may be less than about 300 ng per kg of
body weight. Alternatively the weekly dose of a selective
.kappa.-receptor antagonist may be less than about 200 ng per kg of
body weight, less than about 150 ng per kg of body weight or less
than about 100 ng per kg of body weight. The initial dose may be
larger, followed by smaller maintenance doses. The dose may be
administered as infrequently as weekly or biweekly, or fractionated
into smaller doses and administered daily, semi-weekly, etc, to
maintain an effective dosage level. A suitable daily dosage of a
selective .kappa.-receptor antagonist is less than about 80 ng per
kg of body weight, Alternatively the daily dosage of a selective
.kappa.-receptor antagonist may be less than about 50 ng per kg of
body weight, less than about 25 ng per kg of body weight, or less
than about 20 ng per kg of body weight.
[0026] The dosage of the insulinogenic agent will vary depending on
the condition of the subject or other relevant medical factors that
modify the activity of the insulinogenic agent or the response of
the subject. For example, the specific dose for a particular
patient depends on the severity of glucose intolerance, age, body
weight, general state of health, diet, the timing and mode of
administration, the rate of excretion, and medicaments used in
combination. The initial dose may be larger, followed by smaller
maintenance doses. The dose may be formulated for extended release,
and administered as infrequently as weekly or biweekly, or
fractionated into smaller doses and administered daily,
semi-weekly, weekly, etc. to maintain an effective dosage level.
One of ordinary skill in the art would be able to determine the
appropriate dose of the insulinogenic agent.
[0027] As used in this specification and the appended claims, the
singular forms "a," "an," and "the" include plural referents unless
the content clearly dictates otherwise. It should also be noted
that the term "or" is generally employed in its sense including
"and/or" unless the content clearly dictates otherwise. All
publications, patents and patent applications referenced in this
specification are indicative of the level of ordinary skill in the
art to which this invention pertains. All publications, patents and
patent applications are herein expressly incorporated by reference
to the same extent as if each individual publication or patent
application was specifically and individually indicated by
reference. In case of conflict between the present disclosure and
the incorporated patents, publications and references, the present
disclosure should control.
[0028] It also is specifically understood that any numerical range
recited herein includes all values from the lower value to the
upper value, i.e., all possible combinations of numerical values
between the lowest value and the highest value enumerated are to be
considered to he expressly stated in this application. For example,
if a concentration range is stated as 1% to 50%, it is intended
that values such as 2% to 40%, 10% to 30%, or 1% to 3%, etc., are
expressly enumerated in this specification. For further example, if
a dosage is stated as less than about 250 ng/kg of body weight, it
is intended that values such as 50 to 200 ng per kg of body weight,
and 100 to 200 ng per kg of body weight are expressly enumerated in
this specification. These are only examples of What is specifically
intended.
[0029] The present invention is further explained by the following
examples, which should not be construed by way of limiting the
scope of the present invention.
EXAMPLE 1
Evaluation of Blood Glucose in a Human
[0030] The daily blood glucose profile of a male subject with early
stage, [non-insulin dependent] Type 2 diabetes was monitored during
extended periods of administration of guanidinylated naltrindole
(GNTI). A summary of the blood glucose reading is shown in Table
1.
TABLE-US-00001 TABLE 1 Summary Average, mg/dL 92 Highest Blood
Glucose, mg/dL 127 Lowest Blood Glucose, mg/dL 61 Standard
Deviation, mg/dL 14 Number of Glucose Readings 227 Days Covered 15
Number of Days Without Tests 0 Average Readings Per Day 15.1
Deleted Glucose Readings 0 Control Readings 13 Deleted Control
Readings 0 Average of Recorded Daily Insulin Shots with All Days
Covered: 0.0 with Days with Insulin Records: 0.0
[0031] A weekly dose of about 70 ng/kg was administered. The
subject's blood glucose (BG) levels were measured in mg/dL at time
intervals as shown in Table 2.
TABLE-US-00002 TABLE 2 Early Overall AM Morning Midday Evening
Night Time Range 00:00 05:00 11:00 15:00 19:00 04:59 10:59 14:59
18:59 23:59 Average, mg/dL 92 79 90 92 91 95 Std. Dev. mg/dL 14 3
12 14 13 15 Number of 227 4 65 56 44 58 Readings Very Highs,
>300 Highs. 156-300 In Target, 65-155 98% 100% 97% 98% 100% 97%
Lows, 51-64 2% 3% 2% 3% Very Lows, <51
[0032] FIGS. 1-3 illustrate the blood glucose readings over an
extended period of time, a modal week and a modal day. The target
blood glucose range was 65-155 mg/dL. Table 3 illustrates that 98%
of the readings were within the target range. The other 2% of the
readings fell below the target range and fell between 51-64 mg/dL.
Note that none of these readings were in the hypoglycemic
range.
TABLE-US-00003 TABLE 3 Reading/Range Glucose Ranges, mg/dL Number
Percent Very High (301-601) 0 0% High (156-300) 0 0% Target
(65-155) 222 98% Lows (51-64) 5 2% Very Low (0-50) 0 0%
[0033] FIGS. 1-3 illustrate the subject's blood glucose readings
over an extended period of time, a modal week, and a modal day.
Accordingly, 227 blood glucose readings were taken, averaging 92
mg/dL over a 15 day period. 98% of all readings were between 65 and
127 mg/dL, and 2% between 61 and 65, representing a very narrow
overall spread. These data exemplify glucose dependent insulin
secretion, one of the characteristics of GNTI mediated incretin
targeted treatment of Type 2 diabetes mellitus subjects.
[0034] Table 4 illustrates the blood glucose statistics by day of
week. These data illustrate that once weekly administration of GNTI
aided in maintaining blood glucose levels within the target
range.
TABLE-US-00004 TABLE 4 Blood Glucose Statistics by Day of Week,
mg/dL Weekdays Weekends Mon Tue Wed Thur Fri Sat Sun Average, mg/dL
92 92 92 87 88 94 96 92 92 Std. Dev, mg/dL 15 12 11 17 15 14 15 10
13 Number of 148 79 30 23 28 36 31 32 47 Readings Very Highs,
>300 Highs, 156-300 In Target, 65-155 97 100 100 87 93 100 100
100 100 (as %) Lows, 51-64 3 13 7 (as %) Very Lows, <51
[0035] The weight of the male subject with early stage Type 2
diabetes mellitus was also monitored during extended periods of
administration on GNTI, decreasing from a baseline body mass index
(BMI) of 25 to an average BMI of 22.8, corresponding to a reduction
by about 9%.
EXAMPLE 2
Evaluation of Blood Glucose in Rhesus Monkeys
[0036] A cohort of rhesus monkeys having progressively increasing
degrees of impaired glucose tolerance were monitored following an
oral dose of GNTI of 86 ng/kg of body weight. Baseline readings of
blood glucose (BG), high density lipoprotein C (HDL-C) and
triglycerides (TG) were taken on Day 0. Table 5 shows the baseline
readings on Day 0 and the results of the second reading on Day 8 of
BG, HDL-C and TG.
TABLE-US-00005 TABLE 5 Oral Dose BG HDL-C TG Animal Sequence GNTI
mg/dL mg/dL mg/dL r89163 Baseline 65 64 59 Day 0 86 ng/kg Day 8 63
84 60 Change (-3%) (+)31% (--) r98068 Baseline 71 81 <45 Day 0
86 ng/kg Day 8 51 85 <45 Change (-)29% (+)5% (--) r96027
Baseline 74 44 112 Day 0 86 ng/kg Day 8 53 46 89 Change (-)28%
(+)10% (-)10% r00072 Baseline 80 48 <45 Day 0 86 ng/kg Day 8 67
61 <45 Change (-)16% (+)27% (--) r01078 Baseline 81 51 <45
Day 0 86 ng/kg Day 8 56 63 <45 Change (-)30% (+)24% (--)
[0037] The effect of GNTI was most readily seen in the change of
blood glucose and blood lipid levels. Subject r89163 had a baseline
blood glucose level of 65, i.e., a "normal" fasting blood glucose
level for a rhesus monkey. The other 4 subjects of the cohort,
i.e., r98060, r96027, r00072, r01078, had baseline blood glucose
levels ranging from 71-81 and were considered to have impaired
glucose tolerance. On day 8, the subjects having impaired glucose
tolerance showed a decrease in blood glucose ranging from 16%-30%
with an average of 26%. Comparatively, the glucose "normal" subject
(r89163), receiving the same GNTI dose, exhibited a decrease in
blood glucose levels of 3% only, and in a glucose dependent manner,
a characteristic feature of physiological incretin action. The
lipid "normal" subject (r98068) exhibited an increase in HDL-C
lipid levels of only 5%, where the four dyslipidemic subjects
(r89163, r96027, r00072, r01078) exhibited HDL-C increase in the
range of 10-31%, with an average of 23%.
EXAMPLE 3
Development of a Meal Tolerance Test (MTT)
[0038] To test for the effect of physiological first phase insulin
secretion, a meal tolerance test (MTT) was developed. The meal
tolerance test involved oral administration of meal based nutrients
within suitable ratios of carbohydrates, proteins and fats. The
ratios used for the given example were 70% carbohydrates, 8%
protein and 22% fats. Prior to administration of a MTT to subject
r89163, baseline blood glucose, insulin and glucagon readings were
taken as seen in Table 6. Following MTT administration, the same
readings were taken at time points of 3-9 minutes to test for
first-phase insulin secretion and the subsequent reduction in blood
glucose. It was determined that a first-phase insulin secretion
dependent slight increase in insulin concentration appeared,
typically, at 5 minutes post MTT administration, followed by a
corresponding blood glucose decrease at 7 minutes. A slight
decrease in glucagon levels was also coupled with first-phase
insulin secretion at 5 minutes. The MTT serves as a corollary to
the intravenous glucose tolerance test (IVGTT) and better measures
first-phase insulin secretion because oral administration of
nutrients has a more direct effect on the incretin effect.
TABLE-US-00006 TABLE 6 Test Teste Glucagon Points Meal Cal BG
Insulin GIP GLP-1 Glucagon Change T0- Animal Min Drug C/P/F (%)
Mg/dL microU/mL Pg/mL Pmol/L Pg/mL T60/120 R89163 None 70/8/22 T-0
69.8 24.43 506.96 Test Meal Administration T-3 73.3 15.17 506.86
T-4 73.3 28.85 517.64 TP5/Ins. 76.7 34.63 480.54 T-6 76.1 32.8
456.43 TP7/BG 74.3 32.02 430.75 T-8 75.5 24.81 448.98 T-9 75.6
24.73 481.24
EXAMPLE 4
Application of the Meal Tolerance Test
[0039] Table 7 illustrates administration of the MTT to two rhesus
monkeys categorized as metabolically "normal" based on "normal"
fasting blood glucose values. The monkeys were given a diet of
55-60% carbohydrates, 15-25% protein, and 15-30% fat. Readings of
blood glucose, insulin, and glucagon were taken at time points 0,
5, 7, 60 and 120 minutes. Additionally, at the same time points,
the incretin hormones, GIP and GLP-1 were measured to more directly
measure the incretin effect. In subject r98038, a significant
increase in insulin and concurrent drop in glucagon were seen at
time point 5, as would be expected. One hour after MTT
administration very significant increases in insulin, GIP and GLP-1
were seen in subject r98038, again coupled with consecutive
decreases in glucagon. Comparison of blood glucose levels at time
points 0 and 120 minutes illustrates that normal metabolic
functioning has returned the subject's blood glucose levels to
normal within 120 minutes of MTT administration.
[0040] By contrast, subject r91081 appears from its baseline blood
glucose level to be a "normal" metabolic subject. However, the
characteristic first-phase insulin secretion at time point 5
following MTT administration is lacking as is the time point 7
blood glucose decrease. Further analysis showed that at 120 minutes
following MTT administration the blood glucose and glucagon values
were markedly higher than the baseline readings. These results
contradict the results for "normal" subject r98038, and strongly
indicate impaired metabolic control which was not discernable from
the fasting blood glucose value.
TABLE-US-00007 TABLE 7 Test Glucagon Points TestMealCal BG Insulin
GIP GLP-1 Glucagon Change T0- Animal Min Drug C/P/F (%) Mg/dL
microU/mL Pg/mL pmol/L Pg/mL T60/120 r98038 None 74/6/20 TP 0 min
58.15 15.68 <8 12.8 180.79 TP 5 min 70 25.52 <8 13.6 145.5 TP
7 min 71.15 17.16 <8 11.3 243.26 TP 60 min 53.7 108.56 640 26.6
182.7 (+) 1% TP120 min 56.3 43.97 560 34.9 138.22 .sup. (-) 23%
r91081 None 70/8/22 TP 0 min 56.15 24.71 175 196 99.8 TP 5 min
63.05 32.44 160 178 95.4 TP 7 min 64.5 59.48 150 146 90.51 TP 60
min 63.85 111.1 950 141 114.49 (+) 14.7% TP120 min 78.15 202.29 950
124 145.32 (+) 45.6%
EXAMPLE 5
[0041] Table 8 illustrates the effect of GNTI on physiologic
control of glucagon levels. Baseline readings were taken on Day 1
of blood glucose, insulin, glucagon, GIP, and GLP-1 in
metabolically impaired rhesus monkeys as determined through
administration of the MTT. Cumulative in-situ doses of 45 ng/kg and
95 ng/kg were active on day 5 and day 7, respectively, during
administration of the MTT. Readings of blood glucose, insulin,
glucagon, GIP, and GLP-1 were taken at time points 0, 60 minutes
and 120 minutes. As Table 8 illustrates, both dosages resulted in a
marked decrease in glucagon at both time points 60 and 120 in all
subjects tested. Comparison to untreated subject r91081 in Table 7
further demonstrates the marked decrease in glucagon.
TABLE-US-00008 TABLE 8 Test Points Glucagon 0/60/120 GNTI
TestMealCal BG Insulin GIP GLP-1 Glucagon Change T0- Animal Minutes
ng/kg C/P/F (%) mg/dL microU/mL pg/mL pmol/L pg/mL T60/120 r96022
Day 1 Baseline No Drug 64 16 25 13 394 Day 5 TP 0 min 45 ng/kg
73/9/18 51 14 8 11 411 TP 60 min 64 122 650 31 267 -35% TP120 min
67 200 n/a 31 261 -36% Day 7 TP 0 min 95 ng/kg 74/7/19 56 14 30 28
472 TP 60 min 70 100 675 35 240 -49% TP120 min 63 54 675 n/a 271
-43% rh2251 Day 1 Baseline No Drug 76 11 140 13 377 Day 5 TP 0 min
45 ng/kg 73/9/18 83 57 50 34 411 TP 60 min 98 396 640 83 366 -11%
TP120 min 78 249 760 82 249 -39% Day 7 TP 0 min 95 ng/kg 75/6/19 82
71 25 60 487 TP 60 min 106 401 675 n/a 351 -28% TP120 min 95 307
775 223 267 -45% rh2258 Day 1 Baseline No Drug 74 15 n/a 10 316 Day
5 TP 0 min 45 ng/kg 74/8/18 69 23 n/a 19 302 TP 60 min 70 145 320
49 172 -43% TP120 min 76 137 775 79 122 -60% Day 7 TP 0 min 95
ng/kg 74/7/19 73 30 20 67 293 TP 60 min 69 54 75 60 107 -63% TP120
min 86 71 240 77 108 -63%
[0042] While the present invention has now been described and
exemplified with some specificity, those skilled in the art will
appreciate the various modifications, including variations,
additions, and omissions that may be made in what has been
described. Accordingly, it is intended that these modifications
also be encompassed by the present invention and that the
scope.
* * * * *