U.S. patent application number 13/234248 was filed with the patent office on 2012-01-12 for method of stimulating the motility of the gastrointestinal system using ipamorelin.
This patent application is currently assigned to HELSINN THERAPEUTICS (U.S.), INC.. Invention is credited to William R. Mann, Richard Nelson, William J. Polvino.
Application Number | 20120010157 13/234248 |
Document ID | / |
Family ID | 46332068 |
Filed Date | 2012-01-12 |
United States Patent
Application |
20120010157 |
Kind Code |
A1 |
Polvino; William J. ; et
al. |
January 12, 2012 |
Method of stimulating the motility of the gastrointestinal system
using ipamorelin
Abstract
The present invention provides a method of stimulating the
motility of the gastrointestinal system in a subject in need
thereof, wherein the subject suffers from maladies (i.e.,
disorders, diseases, conditions, or drug- or surgery-induced
dysfunction) of the gastrointestinal system, by administering to
the subject a ghrelin mimetic, or pharmaceutically acceptable salt
thereof. The invention also provides a method of treating a
gastrointestinal malady by co-administering a ghrelin mimetic with
a laxative, a H.sub.2 receptor antagonist, a serotonin receptor
agonist, pure or mixed, an antacid, an opioid antagonist, a proton
pump inhibitor, a motilin receptor agonist, dopamine antagonist, a
cholinergic agonist, a cholinesterase inhibitor, somatostatin,
octreotide, or any combination thereof.
Inventors: |
Polvino; William J.; (Tinton
Falls, NJ) ; Nelson; Richard; (Morristown, NJ)
; Mann; William R.; (Sparta, NJ) |
Assignee: |
HELSINN THERAPEUTICS (U.S.),
INC.
Bridgewater
NJ
|
Family ID: |
46332068 |
Appl. No.: |
13/234248 |
Filed: |
September 16, 2011 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12313956 |
Nov 26, 2008 |
8039456 |
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13234248 |
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11203639 |
Aug 12, 2005 |
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12313956 |
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60600959 |
Aug 12, 2004 |
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61008828 |
Dec 21, 2007 |
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Current U.S.
Class: |
514/21.8 |
Current CPC
Class: |
A61K 31/417 20130101;
A61K 31/33 20130101; A61P 1/00 20180101; A61K 31/485 20130101; A61K
38/08 20130101; A61K 45/06 20130101; A61K 31/417 20130101; A61K
2300/00 20130101; A61K 31/485 20130101; A61K 2300/00 20130101; A61K
38/08 20130101; A61K 2300/00 20130101 |
Class at
Publication: |
514/21.8 |
International
Class: |
A61K 38/08 20060101
A61K038/08; A61P 1/02 20060101 A61P001/02 |
Claims
1-18. (canceled)
19. A method of treating gastroparesis in a human subject in need
thereof comprising administering to said subject a therapeutically
effective amount of a compound represented by the structural
Formula I: ##STR00003## or a pharmaceutically acceptable salt
thereof.
20. The method of claim 19, wherein the subject is using an opioid
for post-surgical pain management.
21. The method of claim 19, wherein the subject is using an opioid
for chronic pain management.
22. The method of claim 21, wherein the opioid is selected from the
group consisting of morphine, codeine, oxycodone, hydromorphone,
hydrocodone, methadone, and fentanyl.
23. The method of claim 19, wherein the compound is administered in
a manner in order to achieve an effective plasma concentration.
24. A method of stimulating the motility of the gastrointestinal
system in a human subject suffering from gastroparesis comprising
administering to said subject a therapeutically effective amount of
a compound represented by the structural Formula I: ##STR00004## or
a pharmaceutically acceptable salt thereof.
25. The method of claim 24, wherein the compound is administered in
a manner to achieve an effective plasma concentration.
26. The method of claim 19, consisting essentially of administering
said compound or a pharmaceutically acceptable salt thereof.
27. The method of claim 24, consisting essentially of administering
said compound or a pharmaceutically acceptable salt thereof.
28. The method of claim 19, wherein said administration is via
injection.
29. The method of claim 24, wherein said administration is via
injection.
Description
RELATED APPLICATIONS
[0001] This application is a continuation-in-part of U.S.
application Ser. No. 11/203,639, filed Aug. 12, 2005 which claims
the benefit of U.S. Provisional Application 60/600,959, filed Aug.
12, 2004. This application also claims the benefit of U.S.
Provisional Application 61/008,828, filed Dec. 21, 2007. The entire
contents of each of the aforementioned applications is hereby
incorporated by reference herein.
INCORPORATION BY REFERENCE
[0002] All references cited herein, including patents, patent
applications, and published patent applications, are hereby
incorporated by reference in their entireties, whether or not each
is further individually incorporated by reference.
BACKGROUND OF THE INVENTION
[0003] Gastrointestinal (GI) motility is a coordinated
neuromuscular process that transports nutrients through the
digestive system. C. Scarpignato, "Pharmacological Stimulation of
Gastrointestinal Motility Where We Are And Where Are We Going?"
Dig. Dis., 15: 112 (1997). Impaired (i.e., slowed) motility of the
gastrointestinal system, which can be involved in gastroesophageal
reflux disease, gastroparesis (e.g., diabetic and postsurgical),
irritable bowel syndrome, ileus, and constipation (e.g., diet or
opioid-induced), is one of the largest health care burdens of
industrialized nations. S. D. Feighner et al., "Receptor for
Motilin Identified in the Human Gastrointestinal System," Science,
284: 2184-2188 (Jun. 25, 1999).
[0004] Growth hormone secretagogues (GHS), such as ghrelin and
mimetics thereof, have been reported to stimulate gastrointestinal
motility. However, the specific GHS compounds that have been
studied have pharmacokinetic properties that will not allow them to
be used clinically for the treatment of gastrointestinal motility.
Specifically, ghrelin is a 28-amino acid peptide that is produced
in the stomach. The biologically active form of ghrelin, i.e., the
acylated form, has a serum half-life of only 9-13 minutes (Akamizu
et al. (2004) European Journal of Endocrinology 150:447-55).
Additionally, synthetic GHS compounds such as GHRP-6 have been
evaluated for the ability to treat GI motility. Similar to ghrelin,
GHRP-6 has a short serum half life that prohibits the use of this
compound from being used to treat GI motility disorders. Bowers et
al. demonstrated that the serum half life of GHRP-6 is only 20
minutes ((1992) Journal of Clinical Endocrinology and Metabolism
74:292-8).
[0005] In view of the above, an effective, physiological way to
effectively stimulate motility of the gastrointestinal system is
highly desirable and would be an advance in the art.
SUMMARY OF THE INVENTION
[0006] The present invention relates to a method of stimulating the
motility of the gastrointestinal system in a subject in need
thereof, wherein the subject suffers from maladies (i.e.,
disorders, diseases, conditions, or drug- or surgery-induced
dysfunction) of the gastrointestinal system. The method comprises
administering to a subject in need thereof a therapeutically
effective amount of a ghrelin mimetic compound or a
pharmaceutically acceptable salt, hydrate or solvate thereof. In a
preferred embodiment, the ghrelin mimetic is ipamorelin as
represented by Formula I (see below), or a pharmaceutically
acceptable salt, hydrate or solvate thereof.
[0007] As described above, ghrelin and ghrelin mimetic compounds
have been shown to have limited serum half lives, and are therefore
not suitable for use in treating GI motility disorders. In contrast
to ghrelin and GHRP-6, the serum half life of ipamorelin in humans
has been demonstrated to be between 3 and 6.5 hours. Accordingly,
the instant invention provides methods and therapeutically
effective compositions for stimulating the motility of the
gastrointestinal system.
[0008] Stimulation of gastrointestinal motility is used in a method
of treating opioid-induced gastrointestinal dysfunction, e.g.,
morphine-induced bowel dysfunction or constipation, in a subject in
need thereof comprising administering a therapeutically effective
amount of a ghrelin mimetic compound or a pharmaceutically
acceptable salt, hydrate or solvate thereof. The subject may be
using opiates or opioids for post-surgical pain management or for
chronic pain management. Exemplary opiates and opioids include
morphine, codeine, oxycodone, hydromorphone, hydrocodone,
methadone, fentanyl, and combinations with anti-inflammatory agents
such as acetaminophen or aspirin. A preferred ghrelin mimetic is
ipamorelin as represented by Formula I, or a pharmaceutically
acceptable salt, hydrate or solvate thereof.
[0009] Stimulation of gastrointestinal motility can be used to
treat gastroparesis in a subject in need thereof by administering a
therapeutically effective amount of a ghrelin mimetic compound. A
preferred ghrelin mimetic is ipamorelin as represented by Formula
I, or a pharmaceutically acceptable salt, hydrate, derivative, acid
amide, or solvate thereof.
[0010] In a further embodiment, stimulation of gastrointestinal
motility is used in a method of treating gastroesophageal reflux
disease (GERD) in a subject in need thereof comprising
administering a therapeutically effective amount of a ghrelin
mimetic compound. In a particular embodiment, the ghrelin mimetic
is ipamorelin as represented by Formula I, or a pharmaceutically
acceptable salt, hydrate or solvate thereof. In a particular
embodiment, the gastroesophageal reflux disease is nocturnal
gastroesophageal reflux disease.
[0011] The invention also provides methods for stimulating
gastrointestinal motility for the treatment of irritable bowel
syndrome (IBS) in a subject in need thereof by administering a
therapeutically effective amount of a ghrelin mimetic compound. A
preferred ghrelin mimetic is ipamorelin as represented by Formula
I, or a pharmaceutically acceptable salt, hydrate or solvate
thereof. The irritable bowel syndrome can be
constipation-predominant irritable bowel syndrome or alternating
constipation/diarrhea irritable bowel syndrome.
[0012] The invention also provides methods for stimulating
gastrointestinal motility to treat constipation in a subject in
need thereof by administering a therapeutically effective amount of
a ghrelin mimetic compound. A preferred ghrelin mimetic is
ipamorelin as represented by Formula I, or a pharmaceutically
acceptable salt, hydrate or solvate thereof.
[0013] In one embodiment, stimulation of gastrointestinal motility
is used in a method of treating surgery-induced or related
gastrointestinal dysfunction, e.g. post-operative ileus, in a
subject in need thereof comprising administering a therapeutically
effective amount of a ghrelin mimetic compound. In a particular
embodiment, the ghrelin mimetic is ipamorelin as represented by
Formula I, or a pharmaceutically acceptable salt, hydrate or
solvate thereof.
[0014] A preferred ghrelin mimetic is ipamorelin as represented by
Formula I
(.alpha.-Methylalanine-L-histidine-D-.beta.-(2-naphthyl)-alanine-D-phen-
ylalanine-L-lysinamide or
H-Aib-His-.beta.-(2-naphthyl)-D-Ala-D-Phe-Lys-NH.sub.2):
##STR00001##
or a pharmaceutically acceptable salt, hydrate or solvate
thereof.
[0015] It is noted that in this disclosure and particularly in the
claims and/or paragraphs, terms such as "comprises", "comprised",
"comprising" and the like can have the meaning attributed to it in
U.S. Patent law; e.g., they can mean "includes", "included",
"including", and the like; and that terms such as "consisting
essentially of" and "consists essentially of" have the meaning
ascribed to them in U.S. Patent law, e.g., they allow for elements
not explicitly recited, but exclude elements that are found in the
prior art or that affect a basic or novel characteristic of the
invention.
[0016] These and other embodiments are disclosed or are obvious
from and encompassed by, the following Detailed Description.
BRIEF DESCRIPTION OF THE DRAWINGS
[0017] The following Detailed Description, given by way of example,
but not intended to limit the invention solely to the specific
embodiments described, may best be understood in conjunction with
the accompanying drawings, in which:
[0018] FIG. 1 shows the gastrokinetic efficacy of oral
administration of ipamorelin at 10 and 100 mg/kg in a rat model for
post-operative ileus.
[0019] FIG. 2 shows the efficacy of .i.v. administered ipamorelin
at 0.1, 0.25 or 1.0 mg/kg on a rat model of post-operative
ileus.
[0020] FIG. 3 shows the efficacy of i.v. administered ipamorelin at
0.01, 0.03 and 0.1 mg/kg in a rat model for post-operative
ileus.
[0021] FIG. 4 demonstrates that ghrelin did not accelerate gastric
emptying in the rats administered 4 mg/kg of morphine and that rats
administered saline or ghrelin displayed the same group mean
absorbance of phenol red.
[0022] FIG. 5 shows the gastrokinetic efficacy of intravenous
administration of ipamorelin (referred to as 26-0161 in the figure)
at 0.25, 1.0, 2.5 mg/kg as compared to 10 mg/kg RC-1139 to treat
post-operative ileus in a rat model.
[0023] FIG. 6 shows the reversal of morphine-induced slowing of
gastrointestinal motility in humans by the intravenous
administration of a single dose of ipamorelin at doses of 0.01,
0.03 and 0.06 mg/kg.
[0024] FIG. 7 shows a bar graph comparing the effect of various
ghrelin mimetics on stomach emptying.
[0025] FIG. 8 shows a bar graph comparing the effect of various
ghrelin mimetics on gastrointestinal motility through the small
intestine.
[0026] FIG. 9 shows a bar graph comparing the effect of various
ghrelin mimetics on stomach emptying.
[0027] FIG. 10 shows a bar graph comparing the effect of various
ghrelin mimetics on gastrointestinal motility through the small
intestine (distal distance from pyloric sphincter).
[0028] FIG. 11 shows a bar graph comparing the effect of various
ghrelin mimetics on gastrointestinal motility through the small
intestine (proximal distance from pyloric sphincter).
[0029] FIG. 12A-D show the effect of abdominal surgery on colonic
transit time, fecal pellet output, food intake, and body weight
gain in rat model of post-operative ileus.
[0030] FIG. 13 shows colonic transit time in rats after abdominal
surgery after single dose administration of ipamorelin at 0.1 and 1
mg/kg relative to vehicle and a control (GHRP-6).
[0031] FIG. 14 shows cumulative fecal pellet output at 12 h, 24 h
or 48 h in rats after abdominal surgery after single dose
administration of ipamorelin at 0.1 and 1 mg/kg relative to vehicle
and a standard control (GHRP-6).
[0032] FIG. 15 shows cumulative food intake at different time
points between 3-48 h in rats after abdominal surgery after single
dose administration of ipamorelin at 0.1 and 1 mg/kg relative to
vehicle and a standard control (GHRP-6).
[0033] FIG. 16 shows the effect on body weight at 24 h and 48 h in
rats after abdominal surgery after single dose administration of
ipamorelin at 0.1 and 1 mg/kg relative to vehicle and a standard
control (GHRP-6).
[0034] FIG. 17 shows colonic transit time in rats after abdominal
surgery after multiple doses of 0.01, 0.1 and 1.0 mg/kg ipamorelin
relative to vehicle.
[0035] FIG. 18 shows the effect of fecal pellet output over a 48 h
period in rats after abdominal surgery after multiple doses of
ipamorelin at 0.01, 0.1 and 1.0 mg/kg ipamorelin relative to
vehicle.
[0036] FIG. 19 shows the effect on cumulative food intake over a 48
h period in rats after abdominal surgery after multiple doses of
ipamorelin at 0.01, 0.1 and 1.0 mg/kg ipamorelin relative to
vehicle.
[0037] FIG. 20 shows the effect on body weight gain over a 48 h
period in rats after abdominal surgery after multiple doses of
ipamorelin at 0.01, 0.1 and 1.0 mg/kg ipamorelin relative to
vehicle.
DETAILED DESCRIPTION
[0038] The present invention relates to a method of stimulating the
motility of the gastrointestinal system in a subject in need
thereof, wherein the subject suffers from maladies (i.e., disorders
or diseases or drug- or surgery-induced dysfunction) of the
gastrointestinal system. In certain embodiments, the maladies
include opioid-induced gastrointestinal dysfunction, e.g.,
morphine-induced gastrointestinal dysfunction, constipation,
diabetes-related gastroparesis, gastroesophageal reflux disease
(GERD), irritable bowel syndrome (IBS), or drug- or surgery-induced
gastrointestinal dysfunction, e.g., post-operative ileus. The
method comprises administering to a subject in need thereof a
therapeutically effective amount of a ghrelin mimetic compound or a
pharmaceutically acceptable salt, hydrate or solvate thereof. The
ghrelin mimetic is preferably ipamorelin as represented by Formula
I, or a pharmaceutically acceptable salt, hydrate or solvate
thereof.
Ghrelin Mimetics
[0039] As used herein, the terms "ghrelin mimetic" or "ghrelin
mimetic compound" or "ghrelin agonist" are synonymous with the
historical terms "growth hormone secretagogue," or "growth hormone
secretagogue compound". A ghrelin mimetic or ghrelin agonist refers
to a substance (e.g., a molecule, a compound) which promotes
(induces or enhances) at least one function that is characteristic
of binding to the ghrelin receptor (GRLN). The GRLN receptor has
been previously reported in the literature as the GHS.sub.1a
receptor which reflected its first known attribute--secretion of
growth hormone. The ghrelin receptor is primarily expressed in the
hypothalamus and pituitary. Activation of these receptors in the
pituitary induces the secretion of growth hormone. In addition to
inducing the secretion of growth hormone, recent studies have shown
the ghrelin mimetics can increase appetite and body weight. In a
particular embodiment, the ghrelin mimetics are those described in
U.S. Pat. Nos. 5,767,085, 6,303,620, 6,576,648, 5,977,178,
6,566,337, 6,083,908, 6,274,584 and 6,919,315, the entire content
of all of which are incorporated herein by reference.
[0040] Subsequently the GRLN receptor was identified in locations
in the body other than the pituitary and hypothalamus, such as the
gastrointestinal tract and the vasculature. The binding of ghrelin
or ghrelin mimetics to these receptors resulted in pharmacological
activity other than, or in addition to, growth hormone release.
Specifically, this other pharmacologic activity was an increase in
gastrointestinal prokinetic activity as well as changes in cardiac
function. Thus, the growth hormone secretagogue compounds as they
were previously named are now more generally called ghrelin
mimetics or agonists to represent the wider spectrum of
physiological actions resulting from binding to its receptor
(GRLN).
[0041] Most identified ghrelin mimetics have a core peptide
backbone with differing lengths of the backbone (tri-, tetra-,
penta-, and hexapeptides, as well as macrocyclic). It is also
expected that the different molecular structures will result in
differing affinities for the ghrelin receptor and, therefore, could
produce differing pharmacological outcomes. A priori one cannot
determine which molecule might produce unusual activity or potency
relative to others in a general class. It generally emerges from
scientific investigation with an unusual result or finding.
[0042] A compound having GRLN receptor agonist activity can be
identified and activity assessed by any suitable method. For
example, the binding affinity of a GRLN receptor agonist to the
GRLN receptor can be determined employing receptor binding assays
and growth hormone stimulation can be assessed as described in U.S.
Pat. No. 6,919,315, which is incorporated herein by reference. The
ghrelin mimetics can be obtained from any source, including any
commercial source.
[0043] In the case of this invention the preferred ghrelin mimetic
is ipamorelin as represented by the structural Formula I
(.alpha.-Methylalanine-L-histidine-D-.beta.-(2-naphthyl)-alanine-D-phenyl-
alanine-L-lysinamide or
H-Aib-His-.beta.-(2-naphthyl)-D-Ala-D-Phe-Lys-NH.sub.2):
##STR00002##
[0044] or a pharmaceutically acceptable salt, hydrate, acid, amide,
crystal or solvate thereof.
[0045] The present invention is based in part on the surprising
discovery made by the present inventors that certain particular
ghrelin mimetics, in particular, ipamorelin, have a surprisingly
efficacious and potent stimulatory effect on gastrointestinal
motility. Although ipamorelin is a potent growth hormone
secretagogue, its binding affinity with the GRLN receptor is about
2-3 logs weaker than many other reported ghrelin mimetics.
Co-Administered Substances
[0046] Another aspect of the present invention relates to the
co-administration of one or more substances and a ghrelin mimetic,
e.g., ipamorelin, to treat a gastrointestinal disorder, disease, or
condition. Co-administered can mean the administration of two or
more substances together as a single pharmaceutical composition, or
the administration of two or more substances in a short period of
time, e.g., within seconds of each other to within a day of each
other.
Peripherally Acting Opioid Antagonists
[0047] It is possible to administer peripherally acting opioid
receptor antagonists, such as, for example, methylnaltrexone,
naloxone, naltrexone, nalmefene and alvimopan (ENTEREG.TM.), which
do not cross the blood-brain barrier, to treat opioid-induced side
effects without provoking opioid withdrawal symptoms or reverse
analgesia. (Holzer P., "Opioids and Opioid Receptors in the Enteric
Nervous System: From a Problem in Opioid Analgesia to a Possible
New Prokinetic Therapy in Humans," Neurosci Lett., 361(1-3):192-5
(2004), incorporated herein by reference). As used herein,
peripherally acting opioid antagonists refer to opioid antagonists
that act peripherally (i.e., not centrally, for example, do not act
on the central nervous system).
Proton Pump Inhibitors
[0048] In another aspect, the present invention provides for
co-administration of a ghrelin mimetic, e.g., ipamorelin, and a
proton pump inhibitor for the treatment of gastrointestinal
conditions or maladies. Proton pump inhibitors suppress gastric
acid secretion, the final step of acid production, by specific
inhibition of the H.sup.+K.sup.+-ATPase enzyme system at the
secretory surface of gastric parietal cells. Proton pump inhibitors
include benzimidazole compounds, for example, esomeprazole
(NEXIUM.RTM.), omeprazole (PRILOSEC.TM.), lansoprazole
(PREVACID.TM.), rabeprazole (ACIPHEX.TM.). and pantoprazole
(Protonix.TM.). These proton pump inhibitors contain a sulfinyl
group situated between substituted benzimidazole and pyridine
rings. At neutral pH, esomeprazole, omeprazole, lansoprazole, and
pantoprazole are chemically stable, lipid soluble, weak bases that
are devoid of inhibitory activity. These uncharged weak bases reach
parietal cells from the blood and diffuse into the secretory
canaliculi, where the drugs become protonated and thereby trapped.
The protonated species rearranges to form a sulfenic acid and a
sulfenamide, the latter species capable of interacting with
sulfhydryl groups of H.sup.+K.sup.+-ATPase. Full inhibition occurs
with two molecules of inhibitor per molecule of enzyme. The
specificity of the effects of proton pump inhibitors is believed to
derive from: a) the selective distribution of
H.sup.+K.sup.+-ATPase; b) the requirement for acidic conditions to
catalyze generation of the reactive inhibitor; and c) the trapping
of the protonated drug and the cationic sulfenamide within the
acidic canaliculi and adjacent to the target enzyme. Goodman &
Gilman's The Pharmacological Basis of Therapeutics, 9th Edition,
pp. 901-915 (1996), incorporated herein by reference.
H.sub.2 Receptor Antagonists
[0049] In yet another aspect, the present invention provides for
the co-administration of a ghrelin mimetic, e.g., ipamorelin, and
an H.sub.2 receptor antagonist for the treatment of
gastrointestinal conditions or maladies. H.sub.2 receptor
antagonists competitively inhibit the interaction of histamine with
H.sub.2 receptors. They are highly selective and have little or no
effect on H.sub.1 receptors. Although H.sub.2 receptors are present
in numerous tissues, including vascular and bronchial smooth
muscle, H.sub.2 receptor antagonists interfere remarkably little
with physiological functions other than gastric acid secretion.
H.sub.2 receptor antagonists include, but are not limited to,
nizatidine (AXID.TM.), ranitidine (ZANTAC.TM. and TRITEC.TM.),
famotidine (PEPCID AC.TM.), and cimetidine (TAGAMET.TM.. Goodman
& Gilman's The Pharmacological Basis of Therapeutics, 9th
Edition, pp. 901-915 (1996), incorporated herein by reference.
H.sub.2 receptor antagonists inhibit gastric acid secretion
elicited by histamine, other H.sub.2 agonists, gastrin, and, to a
lesser extent, muscarinic agonists. H.sub.2 receptor antagonists
also inhibit basal and nocturnal acid secretion.
Antacids
[0050] Another aspect of the present invention provides a method
for the co-administration of a ghrelin mimetic, e.g., ipamorelin,
and an antacid for treating a gastrointestinal condition or malady.
For example, compounds of the invention can be co-administered with
antacids to neutralize gastric acid. For instance, aluminum and
magnesium hydroxide (MAALOX.TM. and MYLANTA.TM.) neutralize gastric
acidity, resulting in an increase in pH in the stomach and duodenal
bulb.
Laxatives
[0051] The present invention further provides a method for the
co-administration of a ghrelin mimetic, e.g., ipamorelin, and a
laxative for treating a gastrointestinal condition or malady.
Laxatives come in various forms, including, for example, liquids,
tablets, suppositories, powders, granules, capsules, chewing gum,
chocolate-flavored wafers, and caramels. The basic types of
laxatives are bulk-forming laxatives, lubricant laxatives, stool
softeners (also called emollient laxatives), and stimulant
laxatives.
[0052] Bulk-forming laxatives contain materials, such as cellulose
and psyllium, that pass through the digestive tract without being
digested. In the intestines, these materials absorb liquid and
swell, making the stool soft, bulky, and easier to pass. The bulky
stool then stimulates the bowel to move. Laxatives in this group
include such brands as FIBERCON.RTM., FIBERALL.RTM., and
METAMUCIL.RTM..
[0053] Lubricant laxatives include, for example, mineral oil.
Mineral oil is the most widely used lubricant laxative. Taken by
mouth, the oil coats the stool. This keeps the stool moist and soft
and makes it easier to pass. Lubricant laxatives are often used for
patients who need to avoid straining (e.g., after abdominal
surgery).
[0054] Stool softeners (emollient laxatives) make stools softer and
easier to pass by increasing their moisture content. This type of
laxative does not really stimulate bowel movements, but it makes it
possible to have bowel movements without straining. Stool softeners
are best used to prevent constipation in people who need to avoid
straining, because of recent surgery, for example. Stool-softening
agents include, for example, docusate sodium (COLACE.RTM.,
REGUTOL.RTM., and others), docusate calcium (SURFAK.RTM., DC
SOFTGELS.RTM.) and docusate potassium (DIALOSE.RTM.,
DIOCTO-K.RTM.).
Serotonin Receptor (5-HT) Agonists (Pure or Mixed)
[0055] The present invention also provides a method for the
co-administration of a ghrelin mimetic, e.g., ipamorelin, and a
serotonin receptor agonist, such as a 5-HT.sub.4 agonist, for
treating a gastrointestinal condition or malady. The serotonin
agonists can either be a pure or mixed 5-HT receptor subtype(s), or
mixed with other central nervous system receptors such as
dopamine.
[0056] The 5-HT.sub.4 agonists speed up movement of bowel contents
through the colon and reduce sensitivity to intestinal nerve
stimulation. Suitable serotonin agonists which can be used in
combination with the compounds of the invention include, but not
restricted to, rauwolscine, yohimbine, metoclopramide, prucalopride
and tegaserod (ZELNORM.RTM.). Spiller R., "Serotonergic Modulating
Drugs for Functional Gastrointestinal Diseases," Br J Clin
Pharmacol. 54:11-20 (2002) and U.S. Pat. No. 6,413,988,
incorporated herein by reference.
Motilin Receptor Agonists
[0057] The present invention further provides a method for the
co-administration of a ghrelin mimetic, e.g., ipamorelin, and a
motilin receptor agonist for treating a gastrointestinal condition
or malady. Motilin is a peptide of 22 amino acids which is produced
in the gastrointestinal system of a number of species. Motilin
induces smooth muscle contractions in the stomach tissue of dogs,
rabbits, and humans as well as in the colon of rabbits. Apart from
local gastrointestinal intestinal tissues, motilin and its
receptors have been found in other tissues.
[0058] In addition to motilin, there are other substances which are
agonists of the motilin receptor and which elicit gastrointestinal
emptying. One of those agents is the antibiotic erythromycin.
Studies have shown that erythromycin elicits biological responses
that are comparable to motilin itself and therefore can be useful
in the treatment of diseases such as chronic idiopathic intestinal
pseudo-obstruction and gastroparesis. Weber, F. et al., The
American Journal of Gastroenterology, 88:4, 485-90 (1993),
incorporated herein by reference.
Dopamine Antagonists
[0059] Another aspect of the present invention provides a method
for the co-administration of a ghrelin mimetic, e.g., ipamorelin,
and a dopamine antagonist for treating a gastrointestinal condition
or malady.
[0060] Dopamine antagonists are drugs that bind to, but do not
activate, dopamine receptors thereby blocking the actions of
dopamine or exogenous agonists. This class of drugs includes, but
is not limited to, metoclopramide, domperidone, amisulpride,
clebopride, mosapramine, nemonapride, remoxipride, risperidone,
sulpiride, sultopride and ziprasidone.
Cholinesterase Inhibitors
[0061] The present invention also provides a method for the
co-administration of a ghrelin mimetic, e.g., ipamorelin, and a
cholinesterase inhibitor for treating a gastrointestinal condition
or malady. The term "cholinesterase inhibitor" refers to one or
more agents that prolong the action of acetylcholine by inhibiting
its destruction or hydrolysis by cholinesterase. Cholinesterase
inhibitors are also known as acetylcholinesterase inhibitors.
Examples of cholinesterase inhibitors include, but are not limited
to, edrophonium, neostigmine, neostigmine methylsulfate,
pyridostigmine, tacrine and physostigmine, ambenonium chloride
(MYTELASE.RTM.), edrophonium chloride (TENSILON.RTM.), neostigmine
(PROSTIGMINE.RTM.), piridogstimina (MESTINON.RTM.), distigmine
bromide, eptastigmine, galanthamine, axeclidine, acetylcholine
bromine, acetylcholine chloride, aclatonium napadisilate,
benzpyrinium bromide, carbachol, carponium chloride, cemecarium
bromide, dexpanthenol, diisopropyl paraoxon, echothiophate
chloride, eseridine, furtrethonium, methacholine chloride,
muscarine, oxapropanium idoide, and xanomeline.
Stereochemistry
[0062] Many of the compounds described herein can have one or more
chiral centers and therefore can exist in different enantiomeric
forms. If desired, a chiral carbon can be designated with an
asterisk (*). When bonds to the chiral carbon are depicted as
straight lines in the formulas of the invention, it is understood
that both the (R) and (S) configurations of the chiral carbon, and
hence both enantiomers and mixtures thereof are embraced within the
formula. As is used in the art, when it is desired to specify the
absolute configuration about a chiral carbon, one of the bonds to
the chiral carbon can be depicted as a wedge (bonds to atoms above
the plane) and the other can be depicted as a series or wedge of
short parallel lines is (bonds to atoms below the plane). The
Cahn-Inglod-Prelog system can be used to assign the (R) or (S)
configuration to a chiral carbon.
[0063] When a compound of the present invention has two or more
chiral carbons, it can have more than two optical isomers and can
exist in diastereoisomeric forms. For example, when there are two
chiral carbons, the compound can have up to 4 optical isomers and 2
pairs of enantiomers ((S,S)/(R,R) and (R,S)/(S,R)). The pairs of
enantiomers (e.g., (S,S)/(R,R)) are mirror image stereoisomers of
one another. The stereoisomers which are not mirror-images (e.g.,
(S,S) and (R,S)) are diastereomers. The diastereoisomeric pairs may
be separated by methods known to those skilled in the art, for
example chromatography or crystallization and the individual
enantiomers within each pair may be separated as described above.
The present invention includes each diastereoisomer of such
compounds and mixtures thereof.
Screening
[0064] It is understood that ghrelin mimetic compounds can be
identified, for example, by screening libraries or collections of
molecules using suitable methods. Another source for the compounds
of interest are combinatorial libraries which can comprise many
structurally distinct molecular species. Combinatorial libraries
can be used to identify lead compounds or to optimize a previously
identified lead. Such libraries can be manufactured by well-known
methods of combinatorial chemistry and screened by suitable
methods.
Methods of Treating Gastrointestinal Maladies
[0065] The present invention provides a method of stimulating the
motility of the gastrointestinal system in a subject in need
thereof, wherein the subject suffers from maladies (i.e.,
disorders, diseases, conditions, or drug- or surgery-induced
dysfunction) of the gastrointestinal system. The method comprises
administering to a subject in need thereof a therapeutically
effective amount of a ghrelin mimetic compound or a
pharmaceutically acceptable salt, hydrate or solvate thereof. The
ghrelin mimetic is ipamorelin as represented by Formula I, or a
pharmaceutically acceptable salt, hydrate or solvate thereof.
[0066] As used herein, the term "gastrointestinal maladies" refers
to any disease, disorder, condition, or dysfunction resulting in
impaired gastrointestinal function. For example, the
gastrointestinal malady can be opioid-induced gastrointestinal
dysfunction, e.g., morphine-induced constipation, post-operative
ileus, or gastroparesis.
Constipation
[0067] In another aspect, the invention provides a method of
treating constipation by administering a therapeutically effective
amount of a ghrelin mimetic, e.g., ipamorelin. Constipation is a
condition in which a person has uncomfortable or infrequent bowel
movements. A person with constipation produces hard stools that can
be difficult to pass. The person also can feel as though the rectum
has not been completely emptied. Acute constipation begins suddenly
and noticeably. Chronic constipation, on the other hand, can begin
insidiously and persist for months or years.
[0068] The method of treating constipation of the invention can
further comprise co-administering a ghrelin mimetic, e.g.,
ipamorelin, with a therapeutically effective amount of a laxative.
Suitable laxatives include, but are not limited to, bulk forming
laxatives, lubricant laxatives, stool softeners, or any combination
thereof.
Opioid-Induced Constipation
[0069] The invention provides a method of treating opioid-induced
constipation by administering a therapeutically effective amount of
a ghrelin mimetic, e.g., ipamorelin. Use of opioid analgesics to
relieve chronic pain can cause effects on organs outside the
targets in the central nervous system. For example, opioid action
can slow stomach emptying and inhibit bowel movement. The increased
time of fecal contents in the intestines results in excessive
absorption of water and sodium from fecal contents, resulting in
harder, drier stools and constipation. This effect afflicts
approximately 90% of individuals on analgesic pain killers. For
chronic pain patients on opioid medications, the resulting
constipation can be a dose limiting side-effect. In addition,
analgesics used for post-surgical pain management can cause
opioid-induced constipation. Suitable opioids include, but are not
limited to, morphine, codeine, oxycodone, hydromorphone,
hydrocodone, methadone, fentanyl, and combinations with
anti-inflammatory agents such as acetaminophen or aspirin or any
combination thereof.
[0070] The method of treating opioid-induced constipation can
further comprise co-administering a ghrelin mimetic compound, e.g.,
ipamorelin, with a therapeutically effective amount of a
peripherally acting opioid antagonist, a laxative, or any
combination thereof. Suitable peripherally acting opioid
antagonists include, but are not limited to, methylnaltrexone,
naltrexone, nalmefene, naloxone and alvimopan or any combination
thereof. Suitable laxatives include, but are not limited to bulk
forming laxatives, lubricant laxatives, stool softeners, or any
combination thereof.
Post-Operative Ileus
[0071] The present invention provides a method of treating
post-operative ileus by administering a therapeutically effective
amount of a ghrelin mimetic, e.g., ipamorelin. It is well
established that the motility of the gastrointestinal (GI) tract is
temporarily impaired after surgery. The effect that an abdominal
operation has on gastrointestinal motility is generally referred to
as "post-operative ileus," a term denoting disruption of the normal
coordinated movements of the gut, resulting in failure of the
propulsion of intestinal contents. Ileus has also been defined as a
functional, non-mechanical obstruction of the bowel. The term
"post-operative ileus" refers to delay in normal gastric and
colonic emptying.
[0072] The method of treating post-operative ileus can further
comprise co-administering a ghrelin mimetic, e.g., ipamorelin, with
a therapeutically effective amount of a dopamine antagonist.
Suitable dopamine antagonists include, but are not limited to,
metoclopramide, domperidone, amisulpride, clebopride, mosapramine,
nemonapride, remoxipride, risperidone, sulpiride, sultopride and
ziprasidone, or any combination thereof.
Irritable Bowel Syndrome
[0073] The present invention provides a method of treating
irritable bowel syndrome by administering a therapeutically
effective amount of a ghrelin mimetic, e.g., ipamorelin. Irritable
bowel syndrome (IBS) is a functional disorder effecting motility of
the entire gastrointestinal tract that can produce abdominal pain,
constipation, and/or diarrhea. The impaired movement of the
digestive tract in IBS is not accompanied by a change in physical
structure, such as inflammation or tumors. The symptoms of IBS are
thought to be related to abnormal muscle contractions in any part
of the intestines.
[0074] In this syndrome, the gastrointestinal tract is especially
sensitive to gastrointestinal stimuli. Stress, diet, drugs,
hormones, or minor irritants can cause the gastrointestinal tract
to contract abnormally. There are different types of IBS:
constipation-predominant, diarrhea-predominant and alternating
constipation-predominant/diarrhea-predominant IBS.
[0075] The method of treating IBS may comprise co-administering a
ghrelin mimetic compound, e.g., ipamorelin, with a therapeutically
effective amount of H.sub.2 receptor antagonist; a serotonin 5-HT
agonist; a laxative; or any combination thereof.
[0076] Suitable H.sub.2 receptor antagonists include, but are not
limited to, nizatidine, ranitidine, famotidine, and cimetidine, or
any combination thereof. Suitable central nervous system receptor
agonists include, but are not limited to, rauwolscine, yohimbine,
metoclopramide, tegaserod, or any combination thereof. Suitable
laxatives include, but are not limited to, bulk forming laxatives,
lubricant laxatives, stool softeners, or any combination
thereof.
Gastroesophageal Reflux Disorder
[0077] The invention further provides a method of treating
gastroesophageal reflux disorder by administering a therapeutically
effective amount of a ghrelin mimetic, e.g., ipamorelin.
Gastroesophageal reflux disease (GERD) is a condition in which
gastric stomach contents (e.g., bile salts) back up into the food
pipe (esophagus), causing chronic regurgitation of gastric contents
from the stomach into the lower esophagus. Commonly known as
heartburn, GERD causes esophageal irritation and inflammation.
[0078] For people with GERD, the esophageal sphincter (a
ring-shaped muscle located at the lower end of the esophagus to
prevent stomach contents from going backwards into the esophagus)
can fail to carry out its protective duties. Instead of opening
only when a person is eating or swallowing, it relaxes and allows
digestive juices to reflux into the esophagus and irritate the
esophageal lining.
[0079] Two types of GERD have been identified, upright or daytime
GERD and supine or nocturnal GERD. Nocturnal reflux episodes occur
less frequently, but acid clearance is more prolonged. Nocturnal
reflux can be associated with the complications of GERD, such as
esophageal erosions, ulceration, and respiratory symptoms. An
estimated 17 million Americans currently suffer from heartburn and
other symptoms of GERD.
[0080] The method of treating GERD comprises co-administering a
ghrelin mimetic compound, e.g., ipamorelin, with a therapeutically
effective amount of H.sub.2 receptor antagonist; an antacid; a
proton pump inhibitor; or any combination thereof.
[0081] Suitable H.sub.2 receptor antagonist include, but are not
limited to, nizatidine, ranitidine, famotidine, and cimetidine, or
any combination thereof. Suitable antacids include, but are not
limited to, aluminum and magnesium hydroxide and combinations
thereof. Suitable proton pump inhibitors include, but are not
limited to, esomeprazole (NEXIUM.RTM.), omeprazole, lansoprazole,
pantoprazole, or a combination thereof.
Gastroparesis
[0082] The present invention provides a method of treating
gastroparesis, e.g. diabetic or idiopathic, by administering a
therapeutically effective amount of a ghrelin mimetic, e.g.,
ipamorelin. Gastroparesis, also referred to as delayed gastric
emptying, is a disorder in which the stomach takes too long to
empty its contents. It often occurs in people with type 1 diabetes
mellitus or type 2 diabetes mellitus. Gastroparesis can occur when
nerves to the stomach are damaged or stop working. The vagus nerve
controls the movement of food through the digestive tract. If the
vagus nerve is damaged, the muscles of the stomach and intestines
do not work normally, and the movement of food is slowed or
stopped. Diabetes can damage the vagus nerve if blood glucose
levels remain high over a long period of time. High blood glucose
causes chemical changes in nerves and damages the blood vessels
that carry oxygen and nutrients to the nerves.
[0083] The method of treating gastroparesis can comprise
co-administering a ghrelin mimetic compound, e.g., ipamorelin, with
a therapeutically effective amount of dopamine antagonist. Suitable
dopamine antagonists include, but are not limited to,
metoclopramide, domperidone, amisulpride, clebopride, mosapramine,
nemonapride, remoxipride, risperidone, sulpiride, sultopride and
ziprasidone, or any combination thereof.
[0084] The invention further relates to pharmaceutical compositions
useful for stimulating (i.e., inducing) motility of the
gastrointestinal system. The pharmaceutical composition comprises a
ghrelin mimetic and optionally a pharmaceutically acceptable
carrier. The pharmaceutical composition can comprise a second
amount of a suitable therapeutic agent. A suitable therapeutic
agent can be determined based on the condition being treated in the
subject.
[0085] For example, the pharmaceutical composition can comprise a
first amount of a ghrelin mimetic, e.g., ipamorelin, and a second
amount of a laxative when treating constipation. The pharmaceutical
composition of the present invention can optionally contain a
pharmaceutically acceptable carrier. The ghrelin mimetic and
laxative can each be present in the pharmaceutical composition in a
therapeutically effective amount. In another aspect, said first and
second amount can together comprise a therapeutically effective
amount.
[0086] The pharmaceutical composition can comprise a first amount
of a ghrelin mimetic and a second amount of a H.sub.2 receptor
antagonist. The pharmaceutical composition of the present invention
can optionally contain a pharmaceutically acceptable carrier. The
ghrelin mimetic and H.sub.2 receptor antagonist can each be present
in the pharmaceutical composition in a therapeutically effective
amount. In another aspect, said first and second amount can
together comprise a therapeutically effective amount.
[0087] The pharmaceutical composition can comprise a first amount
of a ghrelin mimetic and a second amount of a serotonin receptor
agonist. The pharmaceutical composition can optionally contain a
pharmaceutically acceptable carrier. The ghrelin mimetic and
serotonin receptor agonist can each be present in the
pharmaceutical composition in a therapeutically effective amount.
The first and second amount can together comprise a therapeutically
effective amount.
[0088] The pharmaceutical composition can comprise a first amount
of a ghrelin mimetic, e.g., ipamorelin, and a second amount of an
antacid. The pharmaceutical composition can optionally contain a
pharmaceutically acceptable carrier. The ghrelin mimetic and
antacid can each be present in the pharmaceutical composition in a
therapeutically effective amount. In another aspect, said first and
second amount can together comprise a therapeutically effective
amount.
[0089] The pharmaceutical composition can comprise a first amount
of a ghrelin mimetic and a second amount of an opioid antagonist.
The pharmaceutical composition can optionally contain a
pharmaceutically acceptable carrier. The ghrelin mimetic and opioid
antagonist can each be present in the pharmaceutical composition in
a therapeutically effective amount. The first and second amount can
together comprise a therapeutically effective amount.
[0090] The pharmaceutical composition can comprise a first amount
of a ghrelin mimetic and a second amount of a proton pump
inhibitor. The pharmaceutical composition can optionally contain a
pharmaceutically acceptable carrier. The ghrelin mimetic and proton
pump inhibitor can each be present in the pharmaceutical
composition in a therapeutically effective amount. The first and
second amount can together comprise a therapeutically effective
amount.
[0091] The pharmaceutical composition can comprise a first amount
of a ghrelin mimetic and a second amount of a motilin receptor
agonist. The pharmaceutical composition can optionally contain a
pharmaceutically acceptable carrier. The ghrelin mimetic and
motilin receptor agonist can each be present in the pharmaceutical
composition in a therapeutically effective amount. The first and
second amount can together comprise a therapeutically effective
amount.
[0092] The pharmaceutical composition can comprise a first amount
of a ghrelin mimetic and a second amount of a dopamine antagonist.
The pharmaceutical can optionally contain a pharmaceutically
acceptable carrier. The ghrelin mimetic and dopamine antagonist can
each be present in the pharmaceutical composition in a
therapeutically effective amount. The first and second amount can
together comprise a therapeutically effective amount.
[0093] The pharmaceutical composition can comprise a first amount
of a ghrelin mimetic and a second amount of a cholinesterase
inhibitor. The pharmaceutical composition can optionally contain a
pharmaceutically acceptable carrier. The ghrelin mimetic and
cholinesterase inhibitor can each be present in the pharmaceutical
composition in a therapeutically effective amount. The first and
second amount can together comprise a therapeutically effective
amount.
[0094] The pharmaceutical composition can comprise a first amount
of a ghrelin mimetic and a second amount of somatostatin. The
pharmaceutical composition can optionally contain a
pharmaceutically acceptable carrier. The ghrelin mimetic and
somatostatin can each be present in the pharmaceutical composition
in a therapeutically effective amount. The first and second amount
can together comprise a therapeutically effective amount.
[0095] The invention further relates to use of a ghrelin mimetic
compound for the manufacture of a medicament for stimulating (i.e.,
inducing) the motility of the gastrointestinal system.
[0096] Subject, as used herein, refers to animals such as mammals,
including, but not limited to, primates (e.g., humans), cows,
sheep, goats, horses, pigs, dogs, cats, rabbits, guinea pigs, rats,
mice or other bovine, ovine, equine, canine, feline, rodent or
murine species. In a preferred embodiment, the mammal is a
human.
[0097] As used herein, treating and treatment refer to stimulating
(e.g., inducing) motility of the gastrointestinal system.
[0098] As used herein, therapeutically effective amount refers to
an amount sufficient to elicit the desired biological response. The
desired biological response is stimulating (e.g., inducing)
motility of the gastrointestinal system. The desired biological
response is stimulating (e.g., inducing) motility of the
gastrointestinal system to treat opioid induced constipation in a
subject in need thereof. The subject may be using opioids for
post-surgical pain management or for chronic pain management.
[0099] The desired biological response is stimulating (e.g.,
inducing) motility of the gastrointestinal system to treat
gastroparesis in a subject in need thereof.
[0100] The desired biological response is stimulating (e.g.,
inducing) motility of the gastrointestinal system to treat
gastroesophageal reflux disease in a subject in need thereof. The
gastroesophageal reflux disease is nocturnal gastroesophageal
reflux disease.
[0101] The desired biological response is stimulating (e.g.,
inducing) motility of the gastrointestinal system to treat
irritable bowel syndrome in a subject in need thereof. The
irritable bowel syndrome is constipation-predominant irritable
bowel syndrome. In yet another embodiment, the irritable bowel
syndrome is constipation/diarrhea irritable bowel syndrome.
[0102] The desired biological response is stimulating (e.g.,
inducing) motility of the gastrointestinal system to treat
constipation in a subject in need thereof.
[0103] The desired biological response is stimulating (e.g.,
inducing) motility of the gastrointestinal system to treat
post-operative ileus in a subject in need thereof.
Pharmaceutical Compositions
[0104] Pharmaceutical compositions suitable for use in the present
invention include compositions wherein the active ingredients are
contained in a therapeutically effective amount to achieve its
intended purpose. It will be appreciated that the unit content of
active ingredient or ingredients contained in an individual dose of
each dosage form need not in itself constitute an effective amount
since the necessary effective amount can be reached by
administration of a plurality of dosage units (such as capsules or
tablets or vials or combinations thereof). In addition, it is
understood that at some dosage levels, an effective amount may not
show any measurable effect until after a week, a month, three
months, or six months of usage. Determination of the effective
amounts is well within the capability of those skilled in the art,
especially in light of the detailed disclosure provided herein. The
specific dose level for any particular user will depend upon a
variety of factors including the age, the physical activity level,
general health, and the severity of the gastrointestinal
malady.
[0105] A therapeutically effective dose also refers to that amount
necessary to achieve the desired effect without unwanted or
intolerable side effects. Toxicity and therapeutic efficacy of a
ghrelin mimetic, e.g., ipamorelin, of the invention can be
determined by standard pharmaceutical procedures in cell cultures
or experimental animals. Using standard methods, the dosage that
shows effectiveness in about 50% of the test population, the
ED.sub.50, may be determined. Similarly, the dosage that produces
an undesirable side effect to 50% of the population, the SD.sub.50,
can be determined. The dose ratio between side effect and
therapeutic effects can be expressed as the therapeutic index and
it can be expressed as a ratio between SD.sub.50/ED.sub.50. Ghrelin
mimetics with high therapeutic indexes are preferred, e.g.,
ipamorelin, i.e., those which are effective at low dosage and which
do not have undesirable side effects, if any, until very high
doses. A preferred therapeutic index is greater than about 3, more
preferably, the therapeutic index is greater than 10, most
preferably the therapeutic index is greater than 25, such as, for
example, greater than 50. Furthermore, ghrelin mimetics that do not
have side effects at any dosage levels are more preferred. Finally,
ghrelin mimetics that are effective at low dosages and do not have
side effects at any dosage levels are most preferred. The exact
formulation, route of administration and dosage can be chosen
depending on the desired effect and can be made by those of skill
in the art.
[0106] In certain embodiments, the ghrelin mimetics are formulated
as pharmaceutically acceptable salts. As used herein, the term
pharmaceutically acceptable salt refers to a salt of a compound to
be administered prepared from pharmaceutically acceptable non-toxic
acids including inorganic acids, organic acids, solvates, hydrates,
or clathrates thereof. Examples of such inorganic acids are
hydrochloric, hydrobromic, hydroiodic, nitric, sulfuric, and
phosphoric. Appropriate organic acids may be selected, for example,
from aliphatic, aromatic, carboxylic and sulfonic classes of
organic acids, examples of which are formic, acetic, propionic,
succinic, camphorsulfonic, citric, fumaric, gluconic, isethionic,
lactic, malic, mucic, tartaric, para-toluenesulfonic, glycolic,
glucuronic, maleic, furoic, glutamic, benzoic, anthranilic,
salicylic, phenylacetic, mandelic, embonic (pamoic),
methanesulfonic, ethanesulfonic, pantothenic, benzenesulfonic
(besylate), stearic, sulfanilic, alginic, galacturonic, and the
like.
[0107] The ghrelin mimetics of the invention can be prepared in the
form of their hydrates, such as hemihydrate, monohydrate,
dihydrate, trihydrate, tetrahydrate and the like and as
solvates.
[0108] The ghrelin mimetics, e.g., ipamorelin, and derivatives
thereof and any co-administered agents can be incorporated into any
suitable pharmaceutical compositions which may be appropriate or
suitable for administration. Such compositions typically comprise
an active agent (e.g., a ghrelin mimetic of the invention) and a
pharmaceutically acceptable carrier.
[0109] As used herein, "pharmaceutically acceptable carrier" is
intended to include any and all solvents, dispersion media,
coatings, antibacterial and antifungal agents, isotonic and
absorption delaying agents, and the like, compatible with
pharmaceutical administration. 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 compound, use thereof in the compositions is contemplated.
Supplementary active compounds can also be incorporated into the
pharmaceutical compositions of the invention. Modifications can be
made to any of the pharmaceutical composition components to affect
solubility or clearance of the factors of the invention. Peptidic
molecules may also be synthesized with D-amino acids to increase
resistance to enzymatic degradation. In some cases, the composition
can be co-administered with one or more solubilizing agents,
preservatives, and permeation enhancing agents.
Administration of Ghrelin Mimetics
[0110] The therapeutically effective amount or dose will depend on
the age, sex and weight of the patient, and the current medical
condition of the patient. The skilled artisan will be able to
determine appropriate dosages depending on these and other factors
to achieve the desired biological response.
[0111] A suitable dose per day for a ghrelin mimetic of the
invention can be in the range of from about 1 ng to about 10,000
mg, about 5 ng to about 9,500 mg, about 10 ng to about 9,000 mg,
about 20 ng to about 8,500 mg, about 30 ng to about 7,500 mg, about
40 ng to about 7,000 mg, about 50 ng to about 6,500 mg, about 100
ng to about 6,000 mg, about 200 ng to about 5,500 mg, about 300 ng
to about 5,000 mg, about 400 ng to about 4,500 mg, about 500 ng to
about 4,000 mg, about 1 .mu.g to about 3,500 mg, about 5 .mu.g to
about 3,000 mg, about 10 .mu.g to about 2,600 mg, about 20 .mu.g to
about 2,575 mg, about 30 .mu.g to about 2,550 mg, about 40 .mu.g to
about 2,500 mg, about 50 .mu.g to about 2,475 mg, about 100 .mu.g
to about 2,450 mg, about 200 .mu.g to about 2,425 mg, about 300
.mu.g to about 2,000, about 400 .mu.g to about 1,175 mg, about 500
.mu.g to about 1,150 mg, about 0.5 mg to about 1,125 mg, about 1 mg
to about 1,100 mg, about 1.25 mg to about 1,075 mg, about 1.5 mg to
about 1,050 mg, about 2.0 mg to about 1,025 mg, about 2.5 mg to
about 1,000 mg, about 3.0 mg to about 975 mg, about 3.5 mg to about
950 mg, about 4.0 mg to about 925 mg, about 4.5 mg to about 900 mg,
about 5 mg to about 875 mg, about 10 mg to about 850 mg, about 20
mg to about 825 mg, about 30 mg to about 800 mg, about 40 mg to
about 775 mg, about 50 mg to about 750 mg, about 100 mg to about
725 mg, about 200 mg to about 700 mg, about 300 mg to about 675 mg,
about 400 mg to about 650 mg, about 500 mg, or about 525 mg to
about 625 mg.
[0112] Other suitable doses per day for a ghrelin mimetic of the
invention include doses of about or greater than 1 ng, about 5 ng,
about 10 ng, about 20 ng, about 30 ng, about 40 ng, about 50 ng,
about 100 ng, about 200 ng, about 300 ng, about 400 ng, about 500
ng, about 1 .mu.g, about 5 .mu.g, about 10 .mu.g, about 20 .mu.g,
about 30 .mu.g, about 40 .mu.g, about 50 .mu.g, about 100 .mu.g,
about 200 .mu.g, about 300 .mu.g, about 400 .mu.g, about 500 .mu.g
(0.5 mg), about 1 mg, about 1.25 mg, about 1.5 mg, about 2.0 mg,
about 2.5 mg, about 3.0 mg, about 3.5 mg, about 4.0 mg, about 4.5
mg, about 5 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg,
about 50 mg, about 100 mg, about 200 mg, about 300 mg, about 400
mg, about 500 mg, about 600 mg, about 625 mg, about 650 mg, about
675 mg, about 700 mg, about 725 mg, about 750 mg, about 775 mg,
about 800 mg, about 825 mg, about 850 mg, about 875 mg, about 900
mg, about 925 mg, about 950 mg, about 975 mg, about 1000 mg, about
1025 mg, about 1050 mg, about 1075 mg, about 1100 mg, about 1125
mg, about 1150 mg, about 1175 mg, about 1200 mg, about 1225 mg,
about 1250 mg, about 1275 mg, about 1300 mg, about 1325 mg, about
1350 mg, about 1375 mg, about 1400 mg, about 1425 mg, about 1450
mg, about 1475 mg, about 1500 mg, about 1525 mg, about 1550 mg,
about 1575 mg, about 1600 mg, about 1625 mg, about 1650 mg, about
1675 mg, about 1700 mg, about 1725 mg, about 1750 mg, about 1775
mg, about 1800 mg, about 1825 mg, about 1850 mg, about 1875 mg,
about 1900 mg, about 1925 mg, about 1950 mg, about 1975 mg, about
2000 mg, about 2025 mg, about 2050 mg, about 2075 mg, about 2100
mg, about 2125 mg, about 2150 mg, about 2175 mg, about 2200 mg,
about 2225 mg, about 2250 mg, about 2275 mg, about 2300 mg, about
2325 mg, about 2350 mg, about 2375 mg, about 2400 mg, about 2425
mg, about 2450 mg, about 2475 mg, about 2500 mg, about 2525 mg,
about 2550 mg, about 2575 mg, about 2600 mg, about 3,000 mg, about
3,500 mg, about 4,000 mg, about 4,500 mg, about 5,000 mg, about
5,500 mg, about 6,000 mg, about 6,500 mg, about 7,000 mg, about
7,500 mg, about 8,000 mg, about 8,500 mg, about 9,000 mg, or about
9,500 mg.
[0113] A suitable dose of the ghrelin mimetic can be in the range
of from about 0.20 mg to about 4000 mg per day, such as from about
1 mg to about 4000 mg, for example, from about 5 mg to about 3000
mg, such as about 10 mg to about 2400 mg per day. The dose can be
administered in a single dosage or in multiple dosages, for example
from 1 to 4 or more times per day. When multiple dosages are used,
the amount of each dosage can be the same or different.
[0114] A suitable dose for an additional therapeutic agent, such
as, for example, a laxative, can be in same range as described
above for the ghrelin mimetic. The dose of ghrelin mimetic and
additional agent can be the same or different. Suitable doses for
the additional agents can be found in the literature.
[0115] The compounds for use in the method of the invention can be
formulated for administration by any suitable route, such as for
oral or parenteral, for example, transdermal, transmucosal (e.g.,
sublingual, lingual, (trans)buccal), vaginal (e.g., trans- and
perivaginally), (intra)nasal and (trans)rectal), subcutaneous,
intramuscular, intradermal, intra-arterial, intravenous,
inhalation, and topical administration.
[0116] In a preferred embodiment, the compounds of the invention
are formulated for intravenous delivery. In another preferred
embodiment, the compounds of the invention are formulated for oral
delivery. Suitable compositions and dosage forms include tablets,
capsules, caplets, pills, gel caps, troches, dispersions,
suspensions, solutions, syrups, granules, beads, transdermal
patches, gels, powders, pellets, magmas, lozenges, creams, pastes,
plasters, lotions, discs, suppositories, liquid sprays, dry powders
or aerosolized formulations.
[0117] It is preferred that the compounds are orally administered.
Suitable oral dosage forms include, for example, tablets, capsules
or caplets prepared by conventional means with pharmaceutically
acceptable excipients such as binding agents (e.g.,
polyvinylpyrrolidone or hydroxypropylmethylcellulose); fillers
(e.g., lactose, microcrystalline cellulose or calcium phosphate);
lubricants (e.g., magnesium stearate, talc or silica);
disintegrates (e.g., sodium starch glycolate); or wetting agents
(e.g., sodium lauryl sulphate). If desired, the tablets can be
coated, e.g., to provide for ease of swallowing or to provide a
delayed release of active, using suitable methods. Liquid
preparation for oral administration can be in the form of
solutions, syrups or suspensions. Liquid preparations (e.g.,
solutions, suspensions and syrups) are also suitable for oral
administration and can be prepared by conventional means with
pharmaceutically acceptable additives such as suspending agents
(e.g., sorbitol syrup, methyl cellulose or hydrogenated edible
fats); emulsifying agent (e.g., lecithin or acacia); non-aqueous
vehicles (e.g., almond oil, oily esters or ethyl alcohol); and
preservatives (e.g., methyl or propyl p-hydroxy benzoates or sorbic
acid).
[0118] Preferably, a pharmaceutical composition comprising a
ghrelin mimetic of the invention is used to treat a
gastrointestinal malady (e.g., a disorder, disease, condition or
injury of the gastrointestinal tract which impairs gastrointestinal
kinetics) by stimulating gastrointestinal kinetics or motility. The
exact formulation, route of administration and dosage can be chosen
depending on the desired effect and can be made by those of skill
in the art.
[0119] Dosage intervals can be determined by experimental testing.
One or more ghrelin mimetics of the invention could be administered
using a regimen which maintains gastrointestinal motility at about
10% above or below normal, about 20% above or below normal, above
50% above or below normal.
[0120] Another suitable administration method is to provide a
ghrelin mimetic of the invention through an implant or a cell line
capable of expressing a ghrelin mimetic so that the implant or cell
line can provide the ghrelin mimetic to a cell of the
gastrointestinal system.
[0121] A pharmaceutical composition of the invention can be
formulated to be compatible with its intended route of
administration.
[0122] Oral administration refers to the administration of a
pharmaceutical composition of the invention via the mouth through
ingestion, or via any other part of the gastrointestinal system
including the esophagus or through suppository administration.
Parenteral administration refers to the delivery of a composition,
such as a composition comprising a ghrelin mimetic by a route other
than through the gastrointestinal tract (e.g., oral delivery). In
particular, parenteral administration may be via intravenous,
subcutaneous, intramuscular or intramedullary (i.e., intrathecal)
injection. The parenteral preparation can be enclosed in ampoules,
disposable syringes or multiple dose vials made of glass or
plastic. Topical administration refers to the application of a
pharmaceutical agent to the external surface of the skin or the
mucous membranes (including the surface membranes of the nose,
lungs and mouth (in which case it may also be a form of oral
administration, such that the agent crosses the external surface of
the skin or mucous membrane and enters the underlying tissues.
Topical administration of a pharmaceutical agent can result in a
limited distribution of the agent to the skin and surrounding
tissues or, when the agent is removed from the treatment area by
the bloodstream, can result in systemic distribution of the
agent.
[0123] In one form of topical administration contemplated by the
invention, the ghrelin mimetic is delivered by transdermal
delivery. Transdermal delivery refers to the diffusion of an agent
across the barrier of the skin. Absorption through intact skin can
be enhanced by placing the active agent in an oily vehicle before
application to the skin (a process known as inunction) and the use
of microneedles. Passive topical administration may consist of
applying the active agent directly to the treatment site in
combination with emollients or penetration enhancers. Another
method of enhancing delivery through the skin is to increase the
dosage of the pharmaceutical agent. The dosage for topical
administration may be increased up to ten, a hundred or a thousand
folds more than dosages administered by other routes.
[0124] Penetrants for transdermal delivery are generally known in
the art, and include, for example, for transmucosal administration,
detergents, bile salts, and fusidic acid derivatives. Transmucosal
administration can be accomplished through the use of nasal sprays
or suppositories. For administration by inhalation, the ghrelin
mimetics of the invention can be delivered in the form of an
aerosol spray from pressured container or dispenser that contains a
suitable propellant, e.g., a gas such as carbon dioxide, or a
nebulizer. For transdermal administration, the ghrelin mimetics of
the invention can be formulated into ointments, salves, gels, or
creams as generally known in the art.
[0125] In addition, the ghrelin mimetics of the invention may be
delivered by nasal or pulmonary methods. The respiratory delivery
of aerosolized medicaments is described in a number of references,
beginning with Gansslen (1925) Klin. Wochenschr. 4:71 and including
Laube et al. (1993) JAMA 269:2106-21-9; Elliott et al. (1987) Aust.
Paediatr. J. 23:293-297; Wigley et al. (1971) Diabetes 20:552-556.
Corthorpe et al. (1992) Pharm Res 9:764-768; Govinda (1959) Indian
J. Physiol. Pharmacol. 3:161-167; Hastings et al. (1992) J. Appl.
Physiol. 73:1310-1316; Liu et al. (1993) JAMA 269:2106-2109; Nagano
et al. (1985) Jikeikai Med. J. 32:503-506; Sakr (1992) Int. J.
Phar. 86:1-7; and Yoshida et al. (1987) Clin. Res. 35:160-166, each
of which are incorporated herein by reference. Pulmonary delivery
of dry powder medicaments is described in U.S. Pat. No. 5,254,330.
A metered dose inhaler is described in Lee and Sciara (1976) J.
Pharm. Sci. 65:567-572. The intrabronchial administration of
recombinant insulin is briefly described in Schlutiter et al.
(Abstract) (1984) Diabetes 33:75 A and Kohler et al. (1987) Atemw.
Lungenkrkh. 13:230-232. Intranasal and respiratory delivery of a
variety of polypeptides are described in U.S. Pat. No. 5,011,678
and Nagai et al. (1984) J. Contr. Rel. 1:15-22.
[0126] Pharmaceutical compositions suitable for injectable use are
known in the art and include, for example, sterile aqueous
solutions (where water soluble) or dispersions and sterile powders
for the extemporaneous preparation of sterile injectable solutions
or dispersion. For intravenous administration of the ghrelin
mimetics of the invention, physiologically acceptable, suitable
carriers include, for example, physiological saline, bacteriostatic
water, Cremophor EL.TM. (BASF, Parsippany, N.J.) or phosphate
buffered saline (PBS).
[0127] Physiologically acceptable carriers maybe any carrier known
in the field as suitable for pharmaceutical (i.e., topical, oral,
and parenteral) application. Suitable pharmaceutical carriers and
formulations are described, for example, in Remington's
Pharmaceutical Sciences (19th ed.) (Genarro, ed. (1995) Mack
Publishing Co., Easton, Pa.).
[0128] Oral compositions generally include a physiologically
acceptable, inert diluent or an edible carrier. They can be
enclosed in gelatin capsules or compressed into tablets. For the
purpose of oral therapeutic administration, the ghrelin mimetics of
the invention can be incorporated with physiological excipients and
used in the form of tablets, troches, or capsules.
[0129] A number of systems that alter the delivery of injectable
drugs can be used to change the pharmacodynamic and pharmacokinetic
properties of therapeutic agents (see, e.g., K. Reddy, 2000, Annals
of Pharmacotherapy 34:915-923). Drug delivery can be modified
through a change in formulation (e.g., continuous-release products,
liposomes) or an addition to the drug molecule (e.g., pegylation).
Potential advantages of these drug delivery mechanisms include an
increased or prolonged duration of pharmacologic activity, a
decrease in adverse effects, and increased patient compliance and
quality of life. Injectable continuous-release systems deliver
drugs in a controlled, predetermined fashion and are particularly
appropriate when it is important to avoid large fluctuations in
plasma drug concentrations. Encapsulating a drug within a liposome
can produce a prolonged half-life and an increased distribution to
tissues with increased capillary permeability (e.g., tumors).
Pegylation provides a method for modification of therapeutic
peptides or proteins to minimize possible limitations (e.g.,
stability, half-life, immunogenicity) associated with the ghrelin
mimetics of the invention.
[0130] In accordance with the invention, one or more ghrelin
mimetics can be formulated with lipids or lipid vehicles (e.g.,
micelles, liposomes, microspheres, protocells, protobionts,
liposomes, coacervates, and the like) to allow formation of
multimers. Similarly, ghrelin mimetics can be multimerized using
pegylation, cross-linking, disulfide bond formation, formation of
covalent cross-links, glycosylphosphatidylinositol (GPI) anchor
formation, or other established methods. The multimerized ghrelin
mimetics can be formulated into a pharmaceutical composition, and
used to increase or enhance their effects.
[0131] The ghrelin mimetics can also be prepared in the form of
suppositories (e.g., with conventional suppository bases such as
cocoa butter and other glycerides) or retention enemas for rectal
delivery.
[0132] The skilled artisan will appreciate that a variety of
techniques are known that can be used to deliver the ghrelin
mimetics of the invention more specifically to local
gastrointestinal tissues, such as, but not limited to, the stomach,
esophagus, small intestine, or colon. The delivery method will
depend on factors such as the tissue of interest, the nature of the
compound to be delivered, and the duration of the treatment.
[0133] In one aspect, the ghrelin mimetics of the invention are
prepared with carriers that will protect the ghrelin mimetics
against rapid elimination from the body, such as a controlled
release formulation, including implants and microencapsulated
delivery systems. These can be prepared according to methods known
to those skilled in the art, for example, as described in U.S. Pat.
No. 4,522,811.
[0134] Having thus described in detail preferred embodiments of the
present invention, it is to be understood that the invention
defined by the above paragraphs is not to be limited to particular
details set forth in the above description as many apparent
variations thereof are possible without departing from the spirit
or scope of the present invention.
EXAMPLES
[0135] The invention will now be further described by way of the
following non-limiting examples.
[0136] The gastrokinetic efficacy of ipamorelin was evaluated in a
rat model of post-operative ileus. In these studies ipamorelin was
administered either orally (10 mg/kg and 100 mg/kg) or via a single
intravenous injection was administered to male rats following
abdominal surgery over a dose range of 0.01 mg/kg to 1.0 mg/kg. The
effect of ipamorelin on gastric emptying relative to control
animals was determined by administration of a dose of phenol red
via oral gavage followed immediately with the dose of
ipamorelin.
Example 1
Gastrokinetic Efficacy of Ipamorelin (10 or 100 mg/kg) in a Rat
Model of Post-Operative Ileus
[0137] This study evaluated the potential gastrokinetic efficacy of
ipamorelin following a single oral administration in a rat model of
post-operative ileus at a dose of 10 or 100 mg/kg.
TABLE-US-00001 Dose Dose level concentration Dose volume No. of
Treatment Group (mg/kg) (mg/mL) (mL/kg) males 1 Vehicle/control 0 0
5 8 2 Ipamorelin 10 2 5 8 3 Ipamorelin 100 20 5 8 * Animals in this
group underwent all surgical procedures but with no cecum
manipulation.
Methods and Experimental Design
[0138] Male Sprague-Dawley CD.RTM. (Crl: CD.RTM. (SD)) rats (Rattus
norvegicus) were received from Charles River Canada Inc. St.
Constant, Quebec, Canada.
Nine days were allowed between receipt of the animals and the start
of treatment to accustom the animals to the laboratory environment.
At the start of treatment, animals were approximately 7 weeks of
age and were in the weight range of 230 g to 254 g.
[0139] Animals were housed individually in stainless steel wire
mesh-bottomed cages equipped with an automatic watering valve. Each
cage was clearly labeled with a color-coded cage card indicating
project, group, animal numbers and sex. Each animal was uniquely
identified. The targeted conditions for animal room environment and
photoperiod were as follows: Temperature 22.+-.3.degree. C.;
Humidity 50.+-.20%; Light Cycle 12 hours light and 12 hours
dark.
[0140] All animals were given free access (except during designated
procedures) to a standard certified pelleted commercial laboratory
diet (PMI Certified Rodent Chow 5002: PMI Nutrition International
Inc.). The diet was controlled and routinely analyzed by the
manufacturer for maximum allowable concentrations of contaminants
(eg, heavy metals, aflatoxins, organophosphates, chlorinated
hydrocarbons and PCBs). Municipal tap water which had been
softened, purified by reverse osmosis and exposed to ultraviolet
light was provided ad libitum (except during designated
procedures). It is considered that there were no known contaminants
in the dietary materials that could have influenced the outcome of
the study.
[0141] The ipamorelin obtained and used in this study was obtained
from Bachem AG (Bubendorf, Switzerland). The vehicle used was 0.9%
sodium chloride for injection (Baxter). The gastrointestinal marker
used to evaluate level stomach emptying was phenol red (Sigma
Aldrich).
[0142] Appropriate amounts of test article were dissolved in 0.9%
Sodium Chloride for Injection USP. The test article formulations
were adjusted between pH 7.4 to 7.5 with 0.1N/1N hydrochloric acid
or 0.1N sodium hydroxide, as required. All dosing formulations were
kept at room temperature, protected from light. The phenol red was
prepared on the day of dosing as a 5 mg/mL solution in deionized
water and was stored at room temperature, protected from light.
Catheterization Surgery
[0143] Each animal received an antibiotic injection of Benzathine
penicillin G and Procaine penicillin G (0.1 mL) intramuscularly on
the day of surgery and again 2 days following surgery. The animals
were anesthetized with isoflurane gas prior to surgery preparation,
which included shaving of the femoral and dorsal exteriorization
sites. The shaved areas were washed with Chlorhexidine gluconate 4%
followed by a liberal application of Povidone iodine 10%. Prior to
surgery and at the end of the surgical procedure, while under
anesthesia, a bland lubricating ophthalmic agent (Tears Naturale
PM) was administered to each eye. Animals were maintained under
isoflurane gas anesthesia throughout the surgical procedure.
[0144] A small incision was made in the right groin region and the
femoral vein was isolated. A small phlebotomy was made in the vein
and a medical grade silicone-based catheter was inserted and the
tip of the catheter was placed in the vena cava at approximately
the level of the kidneys. The catheter was secured in place with an
appropriate suture material and then brought subcutaneously to the
exteriorization point at the nape of the neck. The femoral site was
irrigated with warm (approximately 37.degree. C.) 0.9% Sodium
Chloride Injection, USP. The femoral site was closed with
interrupted mattress sutures and the exteriorization site with a
purse stitch, which was removed in 5-10 days or depending upon
healing results. A topical antibiotic (Polymyxin B, Bacitracin,
Neomycin) was administered to the catheter exteriorization site,
daily until termination, and the femoral site until considered
unnecessary.
[0145] A jacket was placed on the animal to hold the tether system.
The catheter, prefilled with 0.9% Sodium Chloride Injection,
U.S.P., was fed through the tether system and attached to a swivel
secured to the outside of the cage. The upper portion of the swivel
was connected to the infusion pump and all animals were
continuously infused with 0.9% Sodium Chloride Injection, U.S.P. at
a rate of 0.4 mL/h until initiation of treatment.
Surgery to Induce Post-Operative Ileus
[0146] All treatment procedures were replicated over two
consecutive days with approximately equal numbers of animals from
each group treated on each day. Food was withdrawn from animals
overnight prior to surgery. On the day of surgery animals were
anesthetized with isoflurane gas and a bland ophthalmic lubricant
(Tears Naturale PM) was applied to each eye. The animals were
prepared for surgery by shaving of the entire abdominal region. The
shaved area was cleaned and disinfected appropriately prior to
incision. Using a scalpel blade, the abdomen was opened and the
cecum localized. The cecum was exteriorized and manipulated for
approximately one minute (i.e. gently patted between hands in
saline-soaked gauze). Thereafter, the cecum was repositioned and
the surgical site closed using absorbable suture material
(interrupted sutures) and staples. Animals were then returned to
their home cage to permit recovery from the anesthesia.
[0147] Animals assigned to the sham control group (Group 1)
underwent the same treatment procedures, with the exception that
the cecum was exteriorized and repositioned without
manipulation.
Dose Administration
[0148] Dosing commenced on consecutive days with approximately
equal numbers of animals from each group being dosed on each day.
Prior to dosing, the animals were water deprived. The test/control
articles were administered by oral gavage using a syringe and
flexible gavage tube. The animals were dosed immediately following
the oral gavage dose of phenol red. The dose volume was 5 mL/kg
(for both ipamorelin and phenol red administration) and the actual
dose administered was based on the most recent body weight of each
animal.
Gastrointestinal Assessment
[0149] Approximately 30 minutes following the morphine injection,
all animals received 0.4 mL of phenol red by oral gavage.
Approximately 30 minutes following administration of control or
test article, the rats were euthanized. The stomach was then
exposed by laparotomy, quickly ligated at the pylorus and the
cardia and removed. The stomach was cut open and its contents
extracted with 100 mL of 0.1N NaOH. The phenol red content of this
extract was assayed colorimetrically at 558 nm in a
spectrophotometer. Following collection, samples were stored on wet
ice pending transfer for analysis.
Results
[0150] Orally administered following post-operative ileus,
ipamorelin accelerated stomach emptying by approximately 12.4% and
41.6% at 10 and 100 mg/kg, respectively, when compared to the
control animals, without, however, attaining statistical
significance. See FIG. 1.
CONCLUSION
[0151] Ipamorelin administered orally at doses of 10 or 100 mg/kg
accelerated emptying in a dose dependent relationship, in a rat
model of post-operative ileus.
Example 2
Gastrokinetic Efficacy of Intravenous-Administered Ipamorelin (0.1,
0.25 or 1.0 mg/kg) in a Rat Model of Post-Operative Ileus
[0152] In this study ipamorelin was administered as a slow bolus
intravenous injection via an indwelling catheter (over a period of
ca. 100 seconds).
TABLE-US-00002 Dose Dose level concentration Dose volume No. of
Treatment Group (mg/kg) (mg/mL) (mL/kg) males 1 Sham*/ 0 0 5 8
Vehicle 2 Surgery/ 0 0 5 8 Vehicle 3 Surgery/ 0.1 0.02 5 8
ipamorelin 4 Surgery/ 0.25 0.05 5 8 ipamorelin 5 Sugery/ 1.0 0.2 5
8 ipamorelin *Animals in this group underwent all surgical
procedures but with no cecum manipulation.
Methods and Experimental Design
[0153] Male Sprague-Dawley CD.RTM. (Crl: CD.RTM. (SD)) rats (Rattus
norvegicus) were received from Charles River Canada Inc. St.
Constant, Quebec, Canada. Eight days were allowed between receipt
of the animals and the surgical implantation of the catheters to
allow the animals to become acclimated to the physical and
environmental conditions. Dosing of the animals was initiated
approximately one week following surgical implantation of the
catheters to allow appropriate recovery of the animals prior to
treatment. At the start of treatment, animals were approximately
between 10 to 12 weeks of age and were in the weight range of 327 g
to 397 g.
[0154] Animals were housed individually in stainless steel wire
mesh-bottomed cages equipped with an automatic watering valve. Each
cage was clearly labeled with a color-coded cage card indicating
project, group, animal numbers and sex. Each animal was uniquely
identified. The targeted conditions for animal room environment and
photoperiod were as follows: Temperature 22.+-.3.degree. C.;
Humidity 50.+-.20%; Light Cycle 12 hours light and 12 hours
dark.
[0155] All animals were given free access (except during designated
procedures) to a standard certified pelleted commercial laboratory
diet (PMI Certified Rodent Chow 5002: PMI Nutrition International
Inc.). The diet was controlled and routinely analyzed by the
manufacturer for maximum allowable concentrations of contaminants
(eg, heavy metals, aflatoxins, organophosphates, chlorinated
hydrocarbons and PCBs). Municipal tap water which had been
softened, purified by reverse osmosis and exposed to ultraviolet
light was provided ad libitum (except during designated
procedures).
It is considered that there were no known contaminants in the
dietary materials that could have influenced the outcome of the
study.
[0156] The ipamorelin obtained and used in this study was obtained
from Bachem AG. The vehicle used was 0.9% sodium chloride for
injection (Baxter). The gastrointestinal marker used to evaluate
level stomach emptying was phenol red (Sigma Aldrich).
Catheterization Surgery
[0157] Each animal received an antibiotic injection of Benzathine
penicillin G and Procaine penicillin G (0.1 mL) intramuscularly on
the day of surgery and again 2 days following surgery. The animals
were anesthetized with isoflurane gas prior to surgery preparation,
which included shaving of the femoral and dorsal exteriorization
sites. The shaved areas were washed with Chlorhexidine gluconate 4%
followed by a liberal application of Povidone iodine 10%. Prior to
surgery and at the end of the surgical procedure, while under
anesthesia, a bland lubricating ophthalmic agent (Tears Naturale
PM) was administered to each eye. Animals were maintained under
isoflurane gas anesthesia throughout the surgical procedure.
[0158] A small incision was made in the right groin region and the
femoral vein was isolated. A small phlebotomy was made in the vein
and a medical grade silicone-based catheter was inserted and the
tip of the catheter was placed in the vena cava at approximately
the level of the kidneys. The catheter was secured in place with an
appropriate suture material and then brought subcutaneously to the
exteriorization point at the nape of the neck. The femoral site was
irrigated with warm (approximately 37.degree. C.) 0.9% Sodium
Chloride Injection, USP. The femoral site was closed with
interrupted mattress sutures and the exteriorization site with a
purse stitch, which was removed in 5-10 days or depending upon
healing results. A topical antibiotic (Polymyxin B, Bacitracin,
Neomycin) was administered to the catheter exteriorization site,
daily until termination, and the femoral site until considered
unnecessary.
[0159] A jacket was placed on the animal to hold the tether system.
The catheter, prefilled with 0.9% Sodium Chloride Injection,
U.S.P., was fed through the tether system and attached to a swivel
secured to the outside of the cage. The upper portion of the swivel
was connected to the infusion pump and all animals were
continuously infused with 0.9% Sodium Chloride Injection, U.S.P. at
a rate of 0.4 mL/h until initiation of treatment.
Surgery to Induce Post-Operative Ileus
[0160] All treatment procedures were replicated over two
consecutive days with approximately equal numbers of animals from
each group treated on each day. Food was withdrawn from animals
overnight prior to surgery. On the day of surgery animals were
anesthetized with isoflurane gas and a bland ophthalmic lubricant
(Tears Naturale PM) was applied to each eye. The animals were
prepared for surgery by shaving of the entire abdominal region. The
shaved area was cleaned and disinfected appropriately prior to
incision. Using a scalpel blade, the abdomen was opened and the
cecum localized. The cecum was exteriorized and manipulated for
approximately one minute (i.e. gently patted between hands in
saline-soaked gauze). Thereafter, the cecum was repositioned and
the surgical site closed using absorbable suture material
(interrupted sutures) and staples. Animals were then returned to
their home cage to permit recovery from the anesthesia.
[0161] Animals assigned to the sham control group (Group 1)
underwent the same treatment procedures, with the exception that
the cecum was exteriorized and repositioned without manipulation.
Note that due to technical oversight, the cecums of animal Nos.
2001, 2009, 3001 and 4001 were manipulated with sterile water for
injection USP instead of saline soaker gauze. This minor deviation
was considered to have had no impact on the study outcome or upon
the interpretation of the results.
Dose Administration
[0162] Dosing commenced on consecutive days with approximately
equal numbers of animals from each group being dosed on each day.
The test/control articles were administered as a slow bolus
intravenous injection via an indwelling catheter (over a period of
ca. 100 seconds). The animals were dosed immediately following the
oral gavage dose of phenol red. The dose volume was 5 mL/kg (for
both ipamorelin and phenol red administration) and the actual dose
administered was based on the most recent body weight of each
animal.
Gastrointestinal Assessment
[0163] Prior to dosing of the phenol red, the animals were water
deprived. At approximately 30 minutes post-surgery (ileus), animals
received 0.4 mL of phenol red by oral gavage. Animals were then
dosed and approximately 30 minutes later, were euthanized. Upon
euthanasia, the stomach was exposed by laparotomy, quickly ligated
at the pylorus and the cardia removed. The stomach was cut open and
its contents extracted with 100 mL of 0.1N NaOH. The phenol red
content of this extract was assayed colorimetrically at 558 nm in a
spectrophotometer. Following collection, samples were stored on wet
ice pending transfer for analysis.
Results
[0164] Intravenously administered ipamorelin at doses of 0.1, 0.25
and 1.0 mg/kg following surgery accelerated stomach emptying in the
male albino rats when compared to the vehicle control and sham
control animals, although a dose relationship was not observed.
Animals treated with an intravenous dose (0.1, 0.25 or 1.0 mg/kg)
of ipamorelin displayed a reduction in stomach phenol red content
relative to the vehicle control group (85.7%, 95.6% and 92.2%,
respectively). These reductions reached statistical significance at
the 0.25 and 1.0 mg/kg dose levels (p.ltoreq.0.001 and
p.ltoreq.0.01, respectively). See FIG. 2.
[0165] The average stomach phenol red content of the vehicle
control group was similar and not statistically different from that
of the sham control group and may reflect (unexpected) ileus in the
sham control group or an inability to induce detectable ileus in
the surgery control group. Consequently, when expressed relative to
the sham control group, animals treated with intravenous doses
(0.1, 0.25 or 1.0 mg/kg) of ipamorelin displayed reductions in
stomach phenol red content (87.1%, 96.0% and 93.1%, respectively)
that were statistically significant at all dose levels. See FIG.
2.
CONCLUSION
[0166] Ipamorelin administered intravenously to male albino rats at
a dose of 0.1, 0.25 and 1.0 mg/kg following surgery significantly
accelerated stomach emptying when compared to the sham and vehicle
control animals.
Example 3
Gastrokinetic Efficacy of Intravenous-Administered Ipamorelin
(0.01, 0.03 or 0.1 mg/kg) in a Rat Model of Post-Operative
Ileus
[0167] In this study ipamorelin was administered as a slow bolus
intravenous injection via an indwelling catheter (over a period of
ca. 100 seconds) at a dose of 0.01, 0.03 or 0.1 mg/kg.
TABLE-US-00003 Dose Dose level concentration Dose volume No. of
Treatment Group (mg/kg) (mg/mL) (mL/kg) males 1 Non-operated 0 0 5
8 control* 2 Surgery/ 0 0 5 8 Vehicle 3 Surgery/ 0.01 0.02 0.5 8
ipamorelin 4 Surgery/ 0.03 0.02 1.5 8 ipamorelin 5 Sugery/ 0.1 0.02
5 8 ipamorelin *Animals in this group did not undergo surgery for
ileus induction.
[0168] Male Sprague-Dawley CD.RTM. (Crl: CD.RTM. (SD)) rats (Rattus
norvegicus) were used. Seven days were allowed between receipt of
the animals and the surgical implantation of the catheters to allow
the animals to become acclimated to the physical and environmental
conditions. Dosing of the animals was initiated approximately one
week following surgical implantation of the catheters to allow
appropriate recovery of the animals prior to treatment. At the
start of treatment, animals were approximately 10 weeks of age and
were in the weight range of 334 g to 385 g.
Animals were housed individually in stainless steel wire
mesh-bottomed cages equipped with an automatic watering valve. The
targeted conditions for animal room environment and photoperiod
were as follows: Temperature: 22.+-.3.degree. C.; Humidity:
50.+-.20%; Light cycle: 12 hours light and 12 hours dark. All
animals were examined twice daily for mortality and signs of ill
health or reaction to treatment (except on the day of arrival and
necropsy when the animals were examined once). Individual body
weights were measured at randomization and on the day prior to
dosing (for dose calculation purposes only).
Catheterization Surgery
[0169] A small incision was made in the right groin region and the
femoral vein was isolated. A small phlebotomy was made in the vein
and a medical grade silicone-based catheter was inserted and the
tip of the catheter was placed in the vena cava at approximately
the level of the kidneys. The catheter was secured in place with an
appropriate suture material and then brought subcutaneously to the
exteriorization point at the nape of the neck. The femoral site was
irrigated with warm (approximately 37.degree. C.) 0.9% Sodium
Chloride Injection, USP. The femoral site was closed with
interrupted mattress sutures and the exteriorization site with a
purse stitch, which was removed in 5-10 days or depending upon
healing results. A topical antibiotic (Polymyxin B, Bacitracin,
Neomycin) was administered to the catheter exteriorization site,
daily until termination, and the femoral site until considered
unnecessary.
[0170] A jacket was placed on the animal to hold the tether system.
The catheter, prefilled with 0.9% Sodium Chloride Injection,
U.S.P., was fed through the tether system and attached to a swivel
secured to the outside of the cage. The upper portion of the swivel
was connected to the infusion pump and all animals were
continuously infused with 0.9% Sodium Chloride Injection, U.S.P. at
a rate of 0.4 mL/h until initiation of treatment.
Surgery to Induce Post-Operative Ileus
[0171] All treatment procedures were replicated over two
consecutive days with approximately equal numbers of animals from
each group treated on each day. Food was withdrawn from animals
overnight prior to surgery. On the day of surgery animals were
anesthetized with isoflurane gas and a bland ophthalmic lubricant
(Tears Naturale PM) was applied to each eye. The animals were
prepared for surgery by shaving of the entire abdominal region. The
shaved area was cleaned and disinfected appropriately prior to
incision. Using a scalpel blade, the abdomen was opened and the
cecum localized. The cecum was exteriorized and manipulated for
approximately one minute (i.e. gently patted between hands in
saline-soaked gauze). Thereafter, the cecum was repositioned and
the surgical site closed using absorbable suture material
(interrupted sutures) and staples. Animals were then returned to
their home cage to permit recovery from the anesthesia.
[0172] Animals assigned to the non-operated control group (Group 1)
did not undergo surgery to induce post-operative ileus.
Dose Administration
[0173] Dosing commenced on consecutive days with approximately
equal numbers of animals from each group being dosed on each day.
The test/control articles were administered as a slow bolus
intravenous injection via an indwelling catheter. The animals were
dosed immediately following the oral gavage dose of phenol red. The
dose volume was 5 mL/kg for Groups 1, 2 and 5; 0.5 mL/kg for Group
3 and 1.5 mL/kg for Group 4. The actual dose administered was based
on the most recent body weight of each animal.
Gastrointestinal Assessment
[0174] Prior to dosing of the phenol red, the animals were water
deprived. At approximately 30 minutes post-surgery (ileus), animals
received 0.4 mL of phenol red by oral gavage. Animals were then
dosed and approximately 30 minutes later, were euthanized. Upon
euthanasia, the stomach was exposed by laparotomy, quickly ligated
at the pylorus and the cardia removed. The stomach was cut open and
its contents extracted with 100 mL of 0.1N NaOH. The phenol red
content of this extract was assayed colorimetrically at 558 nm in a
spectrophotometer. Following collection, samples were stored on wet
ice pending transfer for analysis.
Results
[0175] Intravenously administered ipamorelin at doses of 0.01, 0.03
and 0.1 mg/kg following surgery accelerated stomach emptying in the
male albino rats when compared to the vehicle control and
non-operated control animals, although a dose relationship was not
observed. Animals treated with intravenous doses (0.01, 0.03 or 0.1
mg/kg) of ipamorelin displayed reductions in stomach phenol red
content relative to the vehicle control group. These reductions
reached statistical significance at all dose levels (p.ltoreq.0.05
at 0.01 and 0.1 mg/kg and p.ltoreq.0.0 at 0.03 mg/kg). See FIG.
3.
[0176] The average stomach phenol red content of the vehicle
control group was similar and not statistically different from that
of the non-operated control group and may reflect an inability to
induce detectable ileus in the surgery control group. Consequently,
when expressed relative to the non-operated control group, animals
treated with intravenous doses (0.01, 0.03 or 0.1 mg/kg) of
ipamorelin displayed reductions in stomach phenol red content that
were statistically significant at all dose levels.
Example 4
Efficacy of Morphine to Induce Gastric Ileus in Rats
[0177] The purpose of this study was to evaluate the potential
efficacy of morphine to induce gastric ileus in rats. Treatment
groups were set up as follows.
TABLE-US-00004 Ghrelin Morphine Ghrelin concen- Dose Dose level
Dose level tration volume No. of Treatment Group (mg/kg) (ug/kg)
(ug/mL) (mL/kg) males 1 Saline 0 0 0 1.5 8 control 2 Morphine + 1*
0 0 1.5 8 saline 3 Morphine + 1* 50 33.33 1.5 8 ghrelin 4 Morphine
+ 4** 0 0 1.5 8 saline 5 Morphine + 4** 50 33.33 1.5 8 ghrelin
*Morphine was administered at a dose volume of 0.1 mL/kg for groups
2-3 **Morphine was administered at a dose volume of 0.4 mL/kg for
groups 4-5
Methods and Experimental Design
[0178] Male Sprague-Dawley CD.RTM. (Crl: CD.RTM. (SD)) rats (Rattus
norvegicus) were received from Charles River Canada Inc. St.
Constant, Quebec, Canada. Subsequent to arrival, all animals were
subjected to a general physical examination by a qualified member
of the veterinary staff to ensure normal health status. All animals
were considered acceptable for use. Nine days were allowed between
receipt of the animals and the start of treatment to accustom the
animals to the laboratory environment. At the start of treatment,
animals were approximately between 7 weeks of age and were in the
weight range of 231 g to 267 g. Please note that the animals body
weights were slightly outside the protocol stated range. This
deviation was considered to have had no impact on the outcome of
the study or upon interpretation of the results.
[0179] Animals were housed individually in stainless steel wire
mesh-bottomed cages equipped with an automatic watering valve. The
targeted conditions for animal room environment and photoperiod
were as follows: Temperature 22.+-.3.degree. C.; Humidity
50.+-.20%; Light Cycle 12 hours light and 12 hours dark.
[0180] All animals were given free access (except during designated
procedures) to a standard certified pelleted commercial laboratory
diet (PMI Certified Rodent Chow 5002: PMI Nutrition International
Inc.). The diet was controlled and routinely analyzed by the
manufacturer for maximum allowable concentrations of contaminants
(eg, heavy metals, aflatoxins, organophosphates, chlorinated
hydrocarbons and PCBs). The results of these analyses are retained
in the scientific archives of PCS-MTL. Municipal tap water which
had been softened, purified by reverse osmosis and exposed to
ultraviolet light was provided ad libitum (except during designated
procedures). Periodic analysis of the water was subcontracted to
management authorized analytical laboratories which were audited by
the Quality Assurance department of PCS-MTL. The results of these
analyses are retained in the scientific archives of PCS-MTL. It is
considered that there were no known contaminants in the dietary
materials that could have influenced the outcome of the study.
[0181] The dose formulations were prepared on the day of dosing.
The appropriate control was dissolved in the vehicle to achieve the
desired concentration. The morphine sulfate solution was used as
supplied. The phenol red was prepared on the day of dosing as a 5
mg/mL solution and stored at room temperature, protected from light
pending use.
Dose Administration
[0182] Morphine (4 mg/kg or 1 mg/kg) was administered once to the
upper dorsum (scapular region) by subcutaneous injection using a
hypodermic needle attached to a syringe. Morphine was administered
approximately 30 minutes prior to the administration of the
control/positive control articles. The dose volume was 0.1 mL/kg
for Groups 2 and 3 and 0.4 mL/kg for Groups 4 and 5. The actual
dose administered was based on the most recent practical body
weight of each animal.
Gastrointestinal Assessment
[0183] Approximately 30 minutes following the morphine injection,
all animals received 0.4 mL of phenol red by oral gavage.
Approximately 30 minutes following administration of control or
test article, the rats were euthanized. The stomach was then
exposed by laparotomy, quickly ligated at the pylorus and the
cardia and removed. The stomach was cut open and its contents
extracted with 100 mL of 0.1N NaOH. The phenol red content of this
extract was assayed colorimetrically at 558 nm in a
spectrophotometer. Following collection, samples were stored on wet
ice pending transfer for analysis.
Results
[0184] Subcutaneously administered at a dose of 1 or 4 mg/kg,
morphine significantly reduced the stomach emptying by
approximately 42% when compared to the control animals, without
however attaining statistical significance. Ghrelin, a potent
prokinetic peptide known to reverse gastric ileus, administered
intravenously at a dose of 50 .mu.g/kg following exposure with 1
mg/kg of morphine, accelerated stomach emptying by approximately
37% when compared to the animals treated with 1 mg/kg and saline,
and reversed the ileus induced by approximately 11%. However,
ghrelin did not accelerate gastric emptying in the rats
administered 4 mg/kg of morphine and both groups (saline vs
ghrelin) displayed the same group mean absorbance of phenol red
(2.99 and 3.01, respectively). See FIG. 4.
CONCLUSION
[0185] Morphine administered subcutaneously to male albino rats at
doses of 1 or 4 mg/kg induced similar level of gastric ileus, in a
non dose-dependent manner. Ghrelin accelerated stomach emptying and
reversed the gastric ileus induced following exposure with 1 mg/kg
of morphine but had no effects on the stomach emptying of the rats
administered doses of 4 mg/kg of morphine.
Example 5
Gastrokinetic Efficacy of Intravenous Injection of Ipamorelin (1.0,
2.5, 10 mg/kg) as Compared to RC-1139 to Treat Post-Operative Ileus
in a Rat Model
[0186] Ipamorelin was evaluated in a rat model of post-operative
ileus in comparison to RC-1139, a ghrelin mimetic with known
gastrokinetic efficacy. In this study each drug was administered as
a single intravenous injection as shown in the table below.
Treatment groups were as follows.
TABLE-US-00005 Dose Dose level concentration Dose volume No. of
Treatment Group (mg/kg) (mg/mL) (mL/kg) males 1 Vehicle/ 0 0 5 8
Control 2 Ipamorelin 1 0.2 5 8 3 Ipamorelin 2.5 0.5 5 8 4
Ipamorelin 10/0.25 2.5/0.05 5 8* 5 RC-1139 10 2 5 8 (reference
article) *Lethality was observed immediately following
administration of a 10 mg/kg dose (n = 2) and consequently the dose
level was reduced to 0.25 mg/kg for all remaining animals in Group
4
[0187] All animals were examined twice daily for mortality and
signs of ill health or reaction to treatment (except on the day of
arrival and necropsy when the animals were examined once).
Individual body weights were measured at randomization and on the
day prior to dosing (for dose calculation purposes only). Two
animals died immediately following administration of ipamorelin at
a dose of 10 mg/kg and consequently all remaining animals from
Group 4 were dosed at 0.25 mg/kg.
Methods and Experimental Design
[0188] Male Sprague-Dawley CD.RTM. (Crl: CD.RTM. (SD)) rats (Rattus
norvegicus) were randomized to treatment groups At least 5 days was
allowed between receipt of the animals and the start of treatment
to accustom the animals to the laboratory environment. At the start
of treatment, animals were approximately 7 weeks of age and were in
the weight range of 205 g to 272 g.
[0189] Prior to the first dose formulation preparation, a trial
preparation was conducted at 2 mg/mL (test article solution) and at
2 mg/mL (reference article solution) to confirm the suitability of
the proposed formulation method.
[0190] For dosing, formulations were prepared by dissolving the
appropriate quantity of test or reference article in 0.9% Sodium
Chloride for Injection USP. The dosing formulations were then pH
adjusted from 7.4 to 7.5 with 0.1N/1N hydrochloric acid or 0.1N
sodium hydroxide, as required. All dosing formulations were
filtered via 0.2 .mu.m PVDF filters prior to use and were kept at
room temperature, protected from light.
[0191] The phenol red was prepared on the day of dosing as a 5
mg/mL solution in deionized water and was stored at room
temperature, protected from light.
Surgery to Induce Post-Operative Ileus
[0192] All treatment procedures were replicated over two
consecutive days with approximately equal numbers of animals from
each group treated on each day. Food was withdrawn from animals
overnight prior to surgery. On the day of surgery animals were
anesthetized with isoflurane gas and a bland ophthalmic lubricant
was applied to each eye. The animals were prepared for surgery by
shaving of the entire abdominal region. The shaved area was cleaned
and disinfected appropriately prior to incision. Using a scalpel
blade, the abdomen was opened and the cecum localized. The cecum
was exteriorized and manipulated for approximately one minute (i.e.
gently patted between hands in saline-soaked gauze). Thereafter,
the cecum was repositioned and the surgical site closed using
absorbable suture material (interrupted sutures) and staples.
Animals were then returned to their home cage to permit recovery
from the anesthesia.
Dose Administration
[0193] Animals were dosed immediately following the oral gavage
dose of phenol red. The test/control articles and reference article
were administered by intravenous injection (given as a slow bolus
injection over a period of ca. 100 seconds) into the tail vein
using a syringe and appropriate gauge needle. The dose volume was 5
mL/kg and the actual dose administered was based on the most recent
body weight of each animal.
Gastrointestinal Assessment
[0194] Prior to dosing of the phenol red, the animals were water
deprived. At approximately 30 minutes post-surgery, animals
received 0.4 mL of phenol red by oral gavage, then dosed with the
test article, and then, approximately 30 minutes later, were
euthanized. Upon euthanasia, the stomach was exposed by laparotomy,
quickly ligated at the pylorus and the cardia and removed. The
stomach was cut open and its contents extracted with 100 mL of 0.1N
NaOH. The phenol red content of this extract was assayed
colorimetrically at 558 nm in a spectrophotometer. Following
collection, samples were stored on wet ice pending transfer for
analysis.
Results
[0195] Ipamorelin administered intravenously at doses of 0.25, 1
and 2.5 mg/kg following induction of a post-operative ileus
accelerated stomach emptying in the male albino rats, when compared
to the control and RC-1139 treated animals, although a monotonic
dose relationship was not observed. See FIG. 5.
[0196] Intravenous doses of 0.25 and 2.5 mg/kg each displayed a
similar level of efficacy with approximately 50 and 60% reductions
in phenol red content, respectively, while approximately 21%
emptying was observed following the 1 mg/kg dose. The stomach
content of the phenol red marker following administration of the 10
mg/kg RC-1139 dose was approximately 16% lower than that of the
control group.
CONCLUSION
[0197] Ipamorelin administered intravenously at doses of 0.25, 1
and 2.5 mg/kg to male albino rats with post-operative ileus
accelerated stomach emptying, when compared to the control and
reference animals.
Example 6
Administration of Ipamorelin in Healthy Volunteers to Reverse a
Morphine-Induced Slowing of Gastric Emptying
INTRODUCTION
[0198] Delay in gastric emptying plays an etiologic role in several
important target indications including post-operative ileus,
opiate-induced bowel dysfunction, and gastroparesis. In addition,
delays in gastric emptying may promote or exacerbate nausea.
Consequently, an agent such as ipamorelin, with demonstrated
ability to promote gastric emptying in animal models (see above
Examples), may serve an important therapeutic role in a variety of
GI disorders categorized by reduced motility.
[0199] Ipamorelin is a ghrelin mimetic. Ghrelin, a 28-amino acid
peptide hormone, is primarily synthesized in the oxyntic gland in
the stomach and to a lesser degree in other organs of the body such
as the kidney and hypothalamus. Among the important physiologic
effects exerted by ghrelin is its ability to modulate gastric
motility and it has demonstrated a strong prokinetic effect (both
upper and lower GI) in a variety of animal species as well as
humans. See Masuda 2000, Asakawa 2001, Tack 2006. Additionally, in
a rat model ghrelin has been shown to resolve gastric postoperative
ileus. See Trudel 2002.
[0200] The prokinetic activity of ghrelin is likely mediated either
by direct effect on the gut or indirectly by the
vagal-cholinergic-muscarinic pathway. It acts locally in the
stomach to stimulate the firing of vagal afferent neurons and
stimulate gastric motility. See Peeters 2003. Efforts have been
underway for many years to exploit the positive effects of ghrelin
in a variety of disorders via the identification and development of
pharmaceutical agents that mimic ghrelin. Ghrelin has an
exceptionally short half-life (approximately 10 minutes) in humans
and consequently has a limited therapeutic potential. Ipamorelin is
a ghrelin mimetic with a half-life of approximately six hours in
humans, available as an intravenous treatment and thus, is suitable
for therapeutic use.
[0201] The present study was designed to employ a well-validated,
clinical pharmacology model (acetaminophen AUC) for assessment of
the effect of ipamorelin on gastric emptying. Ipamorelin was
demonstrated in the examples described above to have potent,
stimulating effects on gastric emptying in a rat model. This study
was intended to extend these findings into humans.
Dosages
[0202] The dose of ipamorelin selected for this study (0.06 mg/kg
IV infused over 15 minutes), was a dose that has been demonstrated
to be safe and well-tolerated in prior Phase 1 studies. Ipamorelin
was formulated as a 0.5 mg/mL sterile solution with 2 equivalents
of acetic acid in normal saline. The sterile solution was further
diluted with normal saline to an appropriate volume for
administration prior to use.
[0203] The dose of morphine selected for this study (0.05 mg/kg IV
bolus) was a dose that is both clinically relevant as an analgesic
dose and has been demonstrated previously to significantly delay
gastric emptying in normal volunteers [Yuan 1998].
[0204] The acetaminophen dose selected for this study (1000 mg oral
suspension) is a standard Over-the-Counter dose of acetaminophen.
This dose has been employed successfully in prior gastric emptying
studies and produces plasma concentrations which are readily
measured (>0.2 .mu.g/mL).
[0205] The study design was a standard, three-period, randomized,
single-dose crossover study.
[0206] This study was carried out with single-dose administrations,
which is appropriate and well-studied in the acetaminophen AUC
model of gastric emptying. All of the three drugs employed in the
present study have terminal half-lives of six hours or less,
consequently a washout interval of 5-8 days will ensure complete
elimination of the drugs from the body.
[0207] This study's objectives included assessing the ability of
intravenous ipamorelin to reverse opiate-induced delay in gastric
emptying as well as assessing the ability of intravenous ipamorelin
to reverse opiate-induced nausea. The inventors predicted that
ipamorelin will reverse opiate-induced delay in gastric emptying as
assessed by plasma acetaminophen absorption: the plasma
acetaminophen AUC.sub.0-60 following ipamorelin administration will
be 50% greater than that following placebo.
Study Design
[0208] The study was conducted as a single-center, double-blind,
randomized, single-dose, three-way crossover investigation. The
study compared the following treatments:
1. Morphine+Ipamorelin;
2. Morphine Control; and
3. Normal Control
[0209] Plasma samples were obtained over the three hours following
acetaminophen administration for determination of acetaminophen AUC
as a measure of gastric emptying. It was anticipated that morphine
would significantly delay gastric emptying [Yuan 1998]; it was
further anticipated that ipamorelin would reverse the observed
delay in emptying. The primary parameter of interest is the early
absorption of acetaminophen as reflected in the plasma AUC over the
first hour following acetaminophen administration (AUC.sub.0-60).
Parameters of additional interest include AUC.sub.0-180, C.sub.MAX,
and T.sub.MAX.
[0210] The study drug was administered via a well-calibrated
infusion pump (e.g., Harvard pump or similar) over a 15 minute
period. Each subject's dose was calculated based on body weight to
a maximum of 100 kg (6 mg). The dosing volume was then diluted to a
total volume of 15 mL using normal saline for injection as the
diluent. The syringe was drawn up with an air bubble (to facilitate
agitation) and the syringe was mixed by gently inverting six
times.
Morphine Administration
[0211] Morphine (1.0 mg/mL)/placebo was administered by slow bolus
(over 30-60 seconds). The infusion catheter was then flushed
immediately with 3-5 mL normal saline.
Acetaminophen Administration
[0212] An acetaminophen suspension (32 mg/mL) was shaken well prior
to administration. The dose to be administered was 31 mL (992 mg).
The subject was then given an additional 150 mL water to drink.
Results
[0213] Treatment with ipamorelin was well-tolerated and the results
showed a reverse in morphine-induced slowing of gastrointestinal
motility in humans. See FIG. 6.
Example 7
Examination of Lower Doses of Ipamorelin in the Morphine-Induced
Delay in Gastric Emptying in Healthy Male Volunteers
[0214] This study was the same design as that of Example 6a but
evaluated lower doses of IV ipamorelin to reverse opiate-induced
delay in gastric emptying. Data are presented for the twenty three
subjects who completed all treatments. The study treatments were:
(1) untreated (saline) control; (2) morphine 0.05 mg/kg IV; and (3)
morphine 0.05 mg/kg+ipamorelin 0.01 mg/kg IV and (4) morphine 0.05
mg/kg+ipamorelin 0.03 mg/kg IV. Acetaminophen elixir was
administered orally in each treatment cycle to permit assessment of
gastric emptying. The treatments were administered in a
single-blind, placebo-controlled, 3-way crossover study with a
washout of 5-8 days between treatments.
[0215] The results show that administration of morphine delayed
gastric emptying as determined by reduced plasma acetaminophen
levels. This effect was reversed by both doses of ipamorelin and
was comparable to that presented in Example 6. Data are shown in
FIG. 6.
Example 8
Comparison of the Effects of Various Ghrelin Mimetics, Including
Ipamorelin, on Gastrointestinal Motility in Rats
[0216] The objective of this study was to evaluate the
pharmacological effects of a series of ghrelin mimetics on
gastrointestinal motility in rats, as measured by charcoal transit,
relative to a commonly used prokinetic agent, metoclopramide, and
control, following a single intravenous infusion of the
experimental agent.
Treatment groups were as follows:
TABLE-US-00006 Dose Dose Dose Group/ Level Concentration Volume
Number of Identification (mg/kg) (mg/mL) (mL/kg) Males 1/Saline
control 0 0 5 8 2/Metoclopramide 10 1 10 8 3/GHRP-6 0.25 0.05 5 8
4/Ghrelin 0.25 0.05 5 8 5/Ipamorelin 0.25 0.05 5 8 6/RC-1139
acetate 0.25.sup.a 0.05 5 8 7/RC-1089 fumarate 0.25.sup.a 0.05 5 8
8/RC-1187 acetate 0.25.sup.a 0.05 5 8 9/RC-1141 acetate 0.25.sup.a
0.05 5 8 .sup.aDoses were corrected for the test articles salt/base
ratio
[0217] Treatment Procedure:
[0218] Each dosing formulation was prepared on the day of dosing.
For the ghrelin mimetics an aliquot of a 4 mg/mL stock solution was
diluted with an appropriate volume of vehicle to achieve the final
desired concentration. The reference article formulations were also
prepared on the day of dosing by mixing the appropriate amount of
reference article with the appropriate volume of vehicle to achieve
the desired final concentration. The saline and metoclopramide (1
mg/mL) were used as supplied.
[0219] Dosing was performed on consecutive days with approximately
equal numbers of animals being dosed on each day. The animals were
deprived of food overnight prior to treatment. Each formulation was
administered by intravenous injection into the tail vein using a
syringe and appropriate gauge needle. The dose volume was 5 mL/kg
or 10 mL/kg (in the case of metoclopramide only).
[0220] Approximately 30 minutes following dosing with the test
article, all animals received 3 mL of activated charcoal by oral
gavage, followed by a 20-minute period in which the animals were
food and water deprived.
[0221] Following the observation period the rats were euthanized,
the abdominal cavity opened, and the stomach and intestines
removed. The presence or absence of charcoal was documented. The
stomachs were weighed (with or without contents) to give an
indication of gastric emptying. The intestines were opened and
extended to their full length. The charcoal was located and the
distances from the pyloric sphincter to the most proximal and
distal traces of charcoal were measured as was the total distance
from the pyloric sphincter to the cecum. Also, the distance
traveled by the charcoal as a percentage of the total length of the
small intestine was measured.
Results
[0222] Metoclopramide significantly increased intestinal motility,
increasing the distal distance traveled by the charcoal meal from
67.3% for the control group to 86.9% for the metoclopramide-treated
group, a 29% increase. In contrast, the stomach charcoal content of
the group of animals administered metoclopramide was not
significantly different from that of the control group.
[0223] Ipamorelin significantly increased stomach emptying relative
to the control group, reducing the amount of charcoal remaining in
the stomach by 66%. Ghrelin and GHRP-6 also significantly reduced
the amount of charcoal remaining in the stomach, by 57% and 64%,
respectively, relative to the control group, but their effects did
not differ significantly from that of ipamorelin.
[0224] Ipamorelin clearly produced a highly significant
(P.ltoreq.0.001) increase in stomach emptying and a tendency
towards an increase in intestinal motility compared to
saline-treated control group animals, supporting the view that
ipamorelin is a potent gastroprokinetic agent. The results of these
experiments are set forth in FIGS. 7 and 8.
Example 9
Comparison of the Effects of Various Ghrelin Mimetics, Including
Ipamorelin, on Gastrointestinal Motility in Rats
[0225] The objective of this study was to evaluate the
pharmacological effects of a series of ghrelin mimetics on
gastrointestinal motility in rats, as measured by charcoal transit,
relative to control, following a single intravenous infusion of the
experimental agent.
Treatment groups were as follows:
TABLE-US-00007 Number Group/ Dose Level Dose Level.sup.a
Concentration Concentration Dose Volume of Identification
(.mu.moles/kg) (mg/kg) (.mu.moles/mL) (mg/mL) (mL/kg) Males
1/Saline control 0 0 0 0 5 14 2/Ghrelin 75 0.2486 15 0.0497 5 14
3/GHRP-6 75 0.0655 15 0.0131 5 14 4/Ipamorelin 75 0.0562 15 0.0113
5 14 5/RC-1139 acetate 75 0.0523 15 0.0105 5 14 .sup.aDoses of
Ipamorelin and RC-1139 were corrected for their respective purity
.sup.bBoth compounds were co-administered via a single formulation
containing the appropriate amount of each compound and dosed as a
single injection.
Treatment Procedure:
[0226] Dosing commenced on consecutive days with approximately
equal numbers of animals from each group being dosed on each day.
The animals were food deprived overnight prior to treatment. Prior
to dosing, the animals were water deprived. The
test/reference/positive control articles were administered by
intravenous injection into the tail vein using a syringe and
appropriate gauge needle. The dose volume was 5 mL/kg and the
actual dose administered was based on the most recent practical
body weight of each animal.
[0227] Approximately 30 minutes following dosing with the
test/reference or positive control article, all animals received 3
mL of activated charcoal by oral gavage, followed by a 20-minute
period during which the animals were food and water deprived.
[0228] At the end of the observation period, the rats were
euthanized and the abdominal cavity was opened and the stomach and
intestines were removed. The presence or absence of charcoal in the
stomach was documented. The stomachs were weighed (with and without
contents) and this was recorded to give an indication of gastric
emptying. The intestines were opened and extended to their full
length. The charcoal was located and the distances from the pyloric
sphincter to the most proximal and distal traces of charcoal were
measured and recorded, as well as the total distance from the
pyloric sphincter to the cecum (all distances were measured in mm).
In addition to the stomachs weights (with and without contents) the
distance traveled by the charcoal as a percentage of the total
length of the small intestine was recorded to give an indication of
the gastric emptying. Any abnormal or unusual clinical signs noted
during the 20-minute observation period were recorded.
Results
[0229] At a single intravenous dose of 75 .mu.moles/kg, ghrelin and
the three ghrelin mimetics tested in this study (ipamorelin,
GHRP-6, and RC-1139 acetate) displayed disparate effects on gastric
emptying and intestinal transit in the male albino rat when
administered independently. Only ipamorelin displayed similar
prokinetic effects to ghrelin on gastric emptying and both
measurements of intestinal transit (proximal and distal distances
traveled by the charcoal from the pyloric sphincter). Ghrelin,
GHRP-6, and ipamorelin resulted in gastric emptying by one of the
two measures (proximal distance traveled by the charcoal from the
pyloric sphincter) of intestinal transit. See FIGS. 9, 10 and
11.
Example 10
Efficacy of Ipamorelin for the Treatment of Postoperative Ileus in
Rats
[0230] The objective of this study was to investigate whether
ipamorelin would accelerate colonic transit in a rat model of POI.
To perform this assessment both single dose and multiple dose
efficacy were evaluated.
Materials and Methods
[0231] Animals: Adult male Sprague-Dawley rats with indwelling
catheters implanted in the right jugular vein were obtained from
Charles River (Wilmington, Mass.). The initial body weight of the
animals was 250-270 g. The catheters were maintained patent during
the acclimation and were used for the dosing of ipamorelin or
vehicle. An additional group of control rats, not subjected to
surgery and drug or vehicle treatment, were purchased with an
indwelling catheter implanted into the proximal colon (1-2 cm from
the cecum) used for infusion of a dye marker measuring colonic
transit. All rats were single-housed under controlled conditions
(25.degree. C., 12 h light/dark cycle) with free access to food and
water. An acclimation period of at least one week was allowed prior
to the experiments.
Induction of post-operative ileus (POI): POI was induced by a
surgical procedure described as "running of the bowel" (Kalff et
al., 1998). Specifically, rats were anesthetized with isoflurane
(2-3%) inhalation, the abdomen was shaved, disinfected and a
midline incision was made to expose the viscera. The small
intestine and the cecum were exteriorized and inspected for 5 min
using cotton applicators soaked in sterile saline. After completing
the inspection, the intestine was covered with gauze soaked in
saline and the abdomen remained open for additional 10 min. To
study colonic transit, 200 .mu.l of a non-absorbable dye marker
(trypan blue in saline) was injected into the proximal colon (1 cm
distal to the cecum). The incision was then closed with silk
sutures. The whole procedure lasted 25-30 min. Surgeries were
always performed at 6:00-8:00 AM and the animals received
ipamorelin or vehicle treatment during the light phase of the
light/dark cycle. Measurement of colonic transit time: Prior to the
experiments, the rats were fasted for 20-22 h with free access to
water. At the end of the surgical procedure the rats received
intracolonic injection of the dye marker. Following the surgery the
rats were placed in clean home cages supplied with pre-weighed food
(Purina rat chow) and water. Colonic transit time was evaluated as
the period between the end of surgery and the appearance of dye in
the fecal pellet. A naive rat not subjected to surgery and drug or
vehicle treatment was studied on each experimental day together
with the rats with POI. The naive rats were equipped with colonic
catheters used to infuse the dye marker into the colon following a
20-22 h fasting. The data collected from these animals served as a
reference to healthy controls. In previous studies rats with POI
showed a significant delay in colonic transit times compared to
naive animals (Zittel et al., 1998; Greenwood-Van Meerveld,
unpublished data). Cumulative fecal output, food intake and body
weight: Fecal pellets were counted and weighed at 3-h intervals and
12-h intervals during the first 48 post-surgery (see experimental
design). The cumulative fecal output was evaluated by adding the
number of pellets throughout a 48-h post-surgical period. Food
intake was recorded at 3-h interval or 12-h intervals according to
the experimental design and was normalized as g/100 g body weight.
The cumulative food intake was calculated for 48 h post-surgery in
both series of experiments. Body weight was measured daily at
8:00-9:00 AM before fasting the animals, on the day of experiment
before the surgery and at 24 h and 48 h post-surgery. Changes in
body weight are expressed as body weight gain compared to the
weight of the fasted animal taken before the surgery. Test and
control articles: The test compound ipamorelin (free base) was
converted to the diacetate salt by mixing with 2 molar equivalents
of glacial acetic acid. Stock solutions of 0.5 mg/ml were prepared
daily in sterile saline plus glacial acetic acid (0.1 .mu.l per ml)
to bring ipamorelin into solution (pH 3-4). Then the solution was
titrated with NaOH to pH 7.0-7.2. Additional dilutions were made in
saline. Sterile saline was used in the vehicle control experiments.
The positive control [D-Lys.sup.3]-GHRP-6 was purchased from
Sigma-Aldrich (St. Louis, Mo.) and dissolved in sterile saline.
Both the test and control articles were administered as a bolus
i.v. infusion via the jugular catheter at a volume of 0.2 ml/100 g
body weight.
[0232] Data and Statistical Analysis: The data expressed as the
mean.+-.SEM for each group. Differences between groups were
assessed for statistical significance by Student's T test, as well
as by one-way or two-way ANOVA followed by Dunnett's or
Bonferroni's test for multiple comparison where appropriate. A
level of p<0.05 was considered significant. Additionally, the
data for the effects of multiple dosing were evaluated using linear
regression analysis to determine significant differences between
the slopes of the lines between treatments.
Experimental Design:
Experimental Series 1: Determine the Acute Effects Induced by a
Single Dose of Ipamorelin in a Rat Model of POI.
[0233] rats acclimated to the facility for 7 days; patency of
catheters maintained [0234] day 0: rats weighed and fasted at
8:00-9:00 AM [0235] day 1: at 6:00-8:00 AM "running of the bowel"
surgery under isoflurane anesthesia [0236] dye infused in to the
proximal colon at end of surgery [0237] single-dose treatment with
ipamorelin or vehicle [0238] observation in home cage (time to
first fecal pellet; fecal output and food intake at 3, 6, 9, and 12
h post-surgery) [0239] day 2: 7:00-8:00 AM body weight, fecal
output, food intake at 24-h post-surgery [0240] day 3: 7:00-8:00 AM
body weight, fecal output, food intake at 48-h post-surgery [0241]
euthanized with overdose of isoflurane
[0242] Naive rats were fasted but not subjected to surgery. Body
weight, colonic transit, fecal pellet output and food intake were
measured at the same time points as in rats with POI.
TABLE-US-00008 Groups (single dose treatmant): POI + vehicle i.v.)
n = 12 POI + ipamorelin 0.1 mg/kg (i.v.) n = 9 POI + ipamorelin 1
mg/kg (i.v.) n = 10 POI + GHRP-6 20 .mu.g/kg (i.v.) n = 8 Naive (no
surgery, no drug) n = 14
Experimental Series 2: Determine the efficacy of multiple doses of
ipamorelin in a rat model of POI. [0243] rats acclimated to the
facility for 7 days; patency of catheters maintained [0244] day 0:
rats weighed and fasted at 8:00-9:00 AM [0245] day 1: at 6:00-8:00
AM "running of the bowel" surgery under isoflurane anesthesia
[0246] dye infused in to the proximal colon at end of surgery
[0247] multiple dosing of ipamorelin or vehicle (1.sup.st dose at
end of surgery, 2.sup.nd, 3.sup.rd and 4.sup.th dose at 3-h
intervals) [0248] observation in home cage (time to first fecal
pellet, fecal output, food intake at 3, 6, 9 and 12 h post-surgery)
[0249] day 2: 7:00-8:00 AM body weight at 24-h post surgery [0250]
multiple dosing of ipamorelin or vehicle (1.sup.st dose after
measuring body weight, 2.sup.nd, 3.sup.rd and 4.sup.th dose at 3-h
intervals) [0251] observation in home cage (fecal output, food
intake at 24, 27, 30, 33 and 36 h post-surgery) [0252] day 3:
7:00-8:00 AM body weight, fecal output, food intake at 48-h post
surgery euthanasia with isoflurane overdose
[0253] Naive rats were fasted but not subjected to surgery. Body
weight, colonic transit, fecal pellet output and food intake were
measured at the same time points as in rats with POI. Note that the
naive animals remained in their home cage and were not handled in
contrast to the rats with POI, which were handled multiple times
during the dosing of ipamorelin or vehicle.
TABLE-US-00009 Groups (multiple dosing): POI + vehicle (i.v.) n = 8
POI + ipamorelin 0.01 mg/kg (i.v.) n = 8 POI + ipamorelin 0.1 mg/kg
(i.v.) n = 8 POI + ipamorelin 1 mg/kg (i.v.) n = 7 Naive (no
surgery, no drug) n = 8
Results
POI in the Rat: Effect of Abdominal Surgery on Colonic Transit
Time, Fecal Pellet Output, Food Intake and Body Weight.
[0254] A comparison between rats subjected to "running of the bowel
"surgery and naive rats demonstrated that surgery results in the
development of POI characterized as a delay in colonic transit
(FIG. 12A), a decrease in fecal pellet output (FIG. 12 B),
decreased food intake (FIG. 12 C) and body weight gain (FIG. 12
D).
[0255] The rat model of POI. (A) Colonic transit time, measured as
the time to the first marked fecal pellet, is significantly delayed
following surgery compared to naive rats. (B) The cumulative fecal
pellet output at 24 h and 48 h post-surgery is reduced compared to
naive rats (C) Cumulative food intake is significantly decreased at
24 h and 48 h post surgery compared to naive rats. (D) Body weight
gain is reduced following surgery compared to naive rats. Data are
mean.+-.SEM from 12 rats with POI and 14 naive rats. Significance
of the differences between rats subjected to surgery and naive rats
was tested using Student t-test: * p<0.05, ** p<0.01, ***
p<0.001.
[0256] Experimental series 1: Determine the acute effects induced
by a single dose treatment with ipamorelin in a rat model of
POI.
[0257] Experiments were performed in rats with POI to investigate
the efficacy of a single-dose treatment with 0.1 mg/kg or 1 mg/kg
ipamorelin. Ipamorelin or the vehicle was administered via i.v.
infusion following the end of the surgery. The time to the first
fecal pellet (FIG. 13), as well as cumulative fecal pellet output
(FIG. 14), food intake (FIG. 15) and body weight gain (FIG. 16)
were measured during the first 48 h post-surgery and the efficacy
of ipamorelin was evaluated by comparing the effects of ipamorelin
to the effect of the vehicle. In addition, the effect of 20 .mu.g
GHPR-6, an agonist of GRLN, i.e., GHS-R.sub.1a was used as a
positive control (Davenport et al., 2005). The results presented in
FIG. 13 demonstrate that a single post-surgical dose of 1 mg/kg
ipamorelin or 20 .mu.g GHRP-6 significantly decreased the colonic
transit time.
[0258] However, neither ipamorelin nor GHRP-6 had a significant
effect on fecal pellet output (FIG. 14) food intake (FIG. 15) or
body weight gain (FIG. 16) during the first 48 h post-surgery.
[0259] In summary, the results obtained in Experimental series 1
demonstrate that a single-dose treatment with 1 mg/kg ipamorelin
given at the end of surgery decreased the time to the first bowel
movement, but did not induce effects on fecal output and food
intake during the 48-h course of recovery in rats with POI.
However, these results are consistent with those expected based on
the reported half-life of ipamorelin in the rat of 30-60 minutes.
(Johansen et. al., 1998).
[0260] Experimental series 2: Determine the efficacy of multiple
doses of ipamorelin in a rat model of POI.
[0261] The dose-response effect of multiple doses of ipamorelin
(0.01, 0.1 or 1 mg/kg i.v.) was investigated in rats with POI. The
results showed that the multiple dosing of 0.1 mg/kg or 1 mg/kg
ipamorelin caused a significant acceleration of colonic transit
compared to the effect of the vehicle (FIG. 17).
[0262] The effects of multiple doses of ipamorelin on fecal pellet
output are presented in FIG. 18. Multiple doses of ipamorelin
induced an increase in cumulative fecal pellet output compared to
the vehicle. The effect reached significance at doses of 0.1 mg/kg
or 1 mg/kg (FIG. 18). Fecal output increased at a higher rate
(lower 1/slope values) in the rats receiving ipamorelin at doses of
0.1 or 1 mg/kg i.v. compared to rats treated with vehicle. At a
dose of 0.01 mg/kg, ipamorelin showed no significant effect. In
addition, ipamorelin induced an increase food intake (FIG. 19). The
increase in food intake induced by the higher doses of ipamorelin
was associated with an increase in body weight. As illustrated in
FIG. 20, the rats receiving the highest dose of 1 mg/kg ipamorelin,
administered according to the multiple dosing paradigm, gained
significantly more body weight during the first 48 h after surgery
compared to the rats treated with the vehicle.
CONCLUSION
[0263] Overall, the results demonstrate that multiple i.v. dosing
of ipamorelin in rats during the first 48-h after surgery improves
colonic transit and food intake and increases body weight gain
suggesting that post-surgical i.v. infusions of ipamorelin may
ameliorate the symptoms in patients with POI.
[0264] Having now fully provided the instant disclosure, it will be
appreciated by those skilled in the art that the same can be
performed within a wide range of equivalent parameters,
concentrations, and conditions without departing from the spirit
and scope of the disclosure and without undue experimentation.
Further, while the disclosure has been described in connection with
specific embodiments thereof, it will be understood that it is
capable of further modifications. This application is intended to
cover any variations, uses, or adaptations of the disclosure
following, in general, the disclosed principles and including such
departures from the disclosure as come within known or customary
practice within the art to which the disclosure pertains and as may
be applied to the essential features hereinbefore set forth.
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