U.S. patent application number 16/795174 was filed with the patent office on 2020-11-05 for fast acting inhibitor of gastric acid secretion.
The applicant listed for this patent is YALE UNIVERSITY. Invention is credited to John P. Geibel, Philipp Kirchhoff.
Application Number | 20200345767 16/795174 |
Document ID | / |
Family ID | 1000004969584 |
Filed Date | 2020-11-05 |
United States Patent
Application |
20200345767 |
Kind Code |
A1 |
Geibel; John P. ; et
al. |
November 5, 2020 |
Fast Acting Inhibitor of Gastric Acid Secretion
Abstract
The present invention relates to the use of pharmaceutically
acceptable zinc salts, preferably water soluble zinc salts alone or
optionally, in combination with one or more of a protein pump
inhibitor (PPI), H2 blocker, anti-H. pylori
antibiotic/antimicrobial, cytoprotective agent or a combination
agent as otherwise described herein for providing fast action with
optional long duration effect in reducing gastric acid secretion,
raising the pH of the stomach during resting phase as well as
decreasing the duration of stomach acid release during a
secretagogue phase and for treating conditions including
gastroesophageal reflux disease (GERD), non-erosive reflux disease
(NERD), Zollinger-Ellison syndrome (ZE disease), ulcer disease, and
gastric cancer, as well as preventing or reducing the likelihood of
ulcer disease. In addition, the present methods are useful for
treating patients who are non-responsive to proton pump inhibitors
(PPI) and as an alternative to traditional therapies or conditions
which are caused by rapid and complete inhibition of secretagogue
induced acid secretion. The present invention also relates to the
use of one or more water soluble zinc salts, administered in
combination with a therapeutic compound or agent (second
therapeutic agent) which may be delivered orally with enhanced
bioavailability (compared to compounds which are administered in
the absence of water soluble zinc salts) or other favorable
benefits. In addition, therapeutic agents which exhibit sensitivity
to low pH may be advantageously orally administered in combination
with an effective amount of at least one water soluble zinc salt.
Compositions according to the present invention exhibit greater
bioavailability of the active agent when formulated in combination
with a water soluble zinc salt in oral dosage form than when
administered with the water soluble zinc salt.
Inventors: |
Geibel; John P.; (Branford,
CT) ; Kirchhoff; Philipp; (Attendorn, DE) |
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Applicant: |
Name |
City |
State |
Country |
Type |
YALE UNIVERSITY |
New Haven |
CT |
US |
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|
Family ID: |
1000004969584 |
Appl. No.: |
16/795174 |
Filed: |
February 19, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16137714 |
Sep 21, 2018 |
10603339 |
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16795174 |
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13899872 |
May 22, 2013 |
10278989 |
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16137714 |
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11881176 |
Jul 26, 2007 |
8512761 |
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13899872 |
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PCT/US07/01950 |
Jan 25, 2007 |
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11881176 |
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60762595 |
Jan 27, 2006 |
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60764834 |
Feb 3, 2006 |
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60850891 |
Oct 11, 2006 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 45/06 20130101;
A61K 33/30 20130101; A61K 31/4439 20130101; A61K 31/315
20130101 |
International
Class: |
A61K 33/30 20060101
A61K033/30; A61K 31/315 20060101 A61K031/315; A61K 45/06 20060101
A61K045/06; A61K 31/4439 20060101 A61K031/4439 |
Claims
1.-114. (canceled)
115. A method of increasing the pH of the gastric juices of the
stomach of a human patient in need of a rapid increase in stomach
pH, said method comprising orally administering to said patient an
effective amount of a composition consisting essentially of zinc
camosine, wherein said pH of said gastric juices in said patient
increases to at least about 3.0 within a period no greater than
about one hour after administration of said composition.
116. The method according to claim 115 wherein said composition
further includes at least one additional zinc salt selected from
the group consisting of zinc acetate, zinc chloride, zinc lactate,
zinc picolinate and zinc tartrate.
117. The method according to claim 115 wherein said pH of said
gastric juices in said patient increases to at least about 3.5
within a period no greater than about 30 minutes after
administration of said composition.
118. The method according to claim 116 wherein said pH of said
gastric juices in said patient increases to at least about 3.5
within a period no greater than about 30 minutes after
administration of said composition.
119. The method according to claim 115 wherein said pH of said
gastric juices in said patient increases to at least 4.0 within a
period no greater than about 20 minutes after administration of
said composition.
120. The method according to claim 119 wherein said composition
further includes at least one zinc salt selected from the group
consisting of zinc acetate, zinc chloride, zinc lactate, zinc
picolinate and zinc tartrate.
121. The method according to claim 115 wherein said zinc camosine
is coadministered with at least one proton pump inhibitor.
122. The method according to claim 116 wherein said zinc camosine
is coadministered with at least one proton pump inhibitor.
123. The method according to claim 121 wherein said proton pump
inhibitor is selected from the group consisting of omeprazole,
esomeprazole, lansoprazole, pantoprazole and rabeprazole.
124. The method according to claim 122 wherein said proton pump
inhibitor is selected from the group consisting of omeprazole,
esomeprazole, lansoprazole, pantoprazole and rabeprazole.
125. A method of reducing the likelihood of an ulcer developing in
a human patient at risk for an ulcer because of elevated acid
release in the stomach of said patient by rapidly increasing pH of
the gastric juices of the stomach in said patient in response to
said acid release comprising orally administering to said patient
at risk an effective amount of a composition consisting essentially
of zinc camosine, wherein the administration of said composition
increases the pH of gastric juices in the stomach of said patient
to at least about 3.0 within a period no greater than about one
hour after administration of said composition.
126. The method according to claim 125 wherein said pH of said
gastric juices in said patient increases to at least about 3.5
within a period no greater than about 30 minutes after
administration of said composition.
127. The method according to claim 125 wherein said pH of said
gastric juices in said patient increases to at least 4.0 within a
period no greater than about 20 minutes after administration of
said composition.
128. The method according to claim 125 wherein said composition
further includes at least one additional zinc salt selected from
the group consisting of zinc acetate, zinc chloride, zinc lactate,
zinc picolinate and zinc tartrate.
129. The method according to claim 125 wherein said zinc salt is
combined with an effective amount of at least one proton pump
inhibitor.
130. The method according to claim 129 wherein said proton pump
inhibitor is selected from the group consisting of omeprazole,
esomeprazole, lansoprazole, pantoprazole, rabeprazole and mixtures
thereof.
131. The method according to claim 128 wherein said zinc salt(s) is
combined with an effective amount of at least one proton pump
inhibitor.
132. The method according to claim 131 wherein said proton pump
inhibitor is selected from the group consisting of omeprazole,
esomeprazole, lansoprazole, pantoprazole, rabeprazole and mixtures
thereof.
133. A method of treating a human patient in need for a disease
state or condition in which elevated release of acid in the stomach
of said patient occurs selected from the group consisting of
gastroesophageal reflux disease, (GERD), non-erosive reflux disease
(NERD), Zollinger-Ellison syndrome (ZE syndrome), ulcer disease and
gastric cancer comprising orally administering to said patient an
effective amount of a composition to rapidly increase pH of the
gastric juices of the stomach in said patient in response to said
release of acid consisting essentially of zinc carnosine, wherein
the administration of said zinc camosine increases the pH of
gastric juices in the stomach of said patient to at least about 3.0
within a period no greater than about one hour after administration
of said composition.
134. The method according to claim 133 wherein said composition
further includes at least one additional zinc salt selected from
the group consisting of zinc acetate, zinc chloride, zinc lactate,
zinc picolinate and zinc tartrate.
135. The method according to claim 133 wherein said zinc carnosine
is coadministered with at least one agent selected from the group
consisting of a proton pump inhibitor, an H2 blocker, a
cytoprotective agent or a mixture of two or more of these
agents.
136. The method according to claim 134 wherein said zinc camosine
and said zinc salt(s) are coadministered with at least one agent
selected from the group consisting of a proton pump inhibitor, an
H2 blocker, a cytoprotective agent or a mixture of two or more of
these agents.
137. The method according to claim 133 wherein said disease state
or condition is GERD, NERD or ZE syndrome.
138. The method according to claim 134 wherein said disease state
or condition is GERD, NERD or ZE syndrome.
139. The method according to claim 133 wherein said pH of said
gastric juices in said patient increases to at least about 3.5
within a period no greater than about 30 minutes after
administration of said zinc salt(s).
140. The method according to claim 136 wherein said H2 blocker is
cimetidine, famotidine, nizatidine, ranitidine or mixtures
thereof.
141. The method according to claim 136 wherein said cytoprotective
agent is bismuth subsalicylate, sucralfate or a mixture
thereof.
142. The method according to claim 136 wherein said mixture of
agents is prevpac.
143. A method of inhibiting vacuolar H.sup.+-ATPase, H.sup.+,
K.sup.+-ATPase or both H.sup.+-ATPase and H.sup.+, K.about.-ATPase
in the stomach of a human patient in need in which elevated release
of acid in the stomach of said patient occurs comprising
administering to said patient an effective amount of a composition
consisting essentially of zinc camosine to rapidly increase pH of
the gastric juices of the stomach in said patient in response to
said release of acid, wherein the administration of said
composition increases the pH of gastric juices in the stomach of
said patient to at least about 3.0 within a period no greater than
about one hour after administration of said composition.
144. The method according to claim 143 wherein said composition
further includes at least one additional zinc salt selected from
the group consisting of zinc acetate, zinc chloride, zinc lactate,
zinc picolinate and zinc tartrate.
145. The method according to claim 143 wherein said pH of said
gastric juices in said patient increases to at least about 3.5
within a period no greater than about 30 minutes after
administration of said zinc gluconate.
146. The method according to claim 143 wherein said zinc gluconate
is coadministered with at least one proton pump inhibitor.
147. The method according to claim 143 wherein said patient does
not effectively respond to proton pump inhibitor therapy.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to the use of pharmaceutically
acceptable zinc salts, preferably water soluble zinc salts alone or
optionally, in combination with one or more of a protein pump
inhibitor (PPI), H2 blocker, anti-H. pylori
antibiotic/antimicrobial, cytoprotective agent or a combination
agent as otherwise described herein for providing fast action with
optional long duration effect in reducing gastric acid secretion,
including acid secretion in the fundus (by inhibiting vacuolar
H.sup.+-ATPase or H.sup.+/K.sup.+-ATPase) and upper body region of
the stomach (by inhibiting H.sup.+/K.sup.+-ATPase), thus raising
the pH of the stomach during resting phase as well as decreasing
the duration of stomach acid release during a secretagogue phase
and for treating conditions including gastroesophogeal reflux
disease (GERD), non-erosive reflux disease (NERD),
Zollinger-Ellison syndrome (ZE disease), ulcer disease, and gastric
cancer, as well as preventing or reducing the likelihood of ulcer
disease. In addition, the present methods are useful for treating
patients who are non-responsive to proton pump inhibitors (PPI) and
as an alternative to traditional therapies or conditions which are
caused by rapid and complete inhibition of secretagogue induced
acid secretion.
[0002] The present invention also relates to the use of one or more
water soluble zinc salts, administered in combination with a
therapeutic compound or agent (second therapeutic agent) which may
be delivered orally with enhanced bioavailability (compared to
compounds which are administered in the absence of water soluble
zinc salts) or other favorable benefits. In addition, therapeutic
agents which exhibit sensitivity to low pH may be advantageously
orally administered in combination with an effective amount of at
least one water soluble zinc salt. Compositions according to the
present invention exhibit greater bioavailability of the active
agent when formulated in combination with a water soluble zinc salt
in oral dosage form than when administered with the water soluble
zinc salt.
RELATED APPLICATIONS
[0003] This application is a divisional application of U.S. patent
application Ser. No. 16/137,714 filed Sep. 21, 2018, which is a
divisional application of U.S. patent application Ser. No.
13/899,782 filed May 22, 2013, which is a continuation application
of U.S. patent application Ser. No. 11/881,176, filed Jul. 26,
2007, which is a continuation in part application of patent
application PCT/US07/01950, entitled "Fast Acting Inhibitor of
Gastric Acid Secretions", filed Jan. 25, 2007, which claims the
benefit of priority of U.S. provisional application no. U.S.
60/762,595, filed Jan. 27, 2006, U.S. 60/764,834, filed Feb. 3,
2006 and U.S. 60/850,891, filed Oct. 11, 2006, each of said
applications being incorporated by reference herein in their
entirety.
BACKGROUND OF THE INVENTION
[0004] The generation of concentrated 0.16N hydrochloric acid by
the mammalian parietal cell involves a complex combination of
neuronal and hormonal regulatory feedback loops.sup.1-3. Following
activation of the cell there is a complex cellular transfer of ions
that allows for the formation of acid.sup.4-7. A disruption in any
of these components (secretory receptors, or ion transporters) can
lead to either a cessation in the secretion of acid, or in the
hypersecretion of acid. In the latter over 30 million patients per
year suffer from symptoms of acid related diseases with the numbers
increasing yearly.sup.8-11. Clinically the uncontrolled release or
the continued hypersecretion of acid can lead to changes in both
gastric and intestinal epithelium, but can in more serious cases
lead to erosions of the esophagus that can result in metaplasia and
death.sup.12-14. Recent evidence has also emerged that prolonged
recurrent periods of hypersecretory states can lead to gastric
carcinoid formation.sup.15.
[0005] In an attempt to design therapies to prevent hyperacid
secretion a variety of approaches have been employed in recent
years with two of the most successful being: a) inhibition of the
Histamine receptor on the basolateral membrane of the parietal
cell, b) proton pump specific drugs targeted against the
H.sup.+,K.sup.+-ATPase (the so called proton pump inhibitors;
PPI).sup.16-18. Both of these therapies have greatly improved the
quality of life for patients suffering from this disease, however
there is an ever increasing number of patients that have
experienced recurrent disease while still taking the
drugs.sup.19,20. Despite their high degree of efficacy and
worldwide clinical use, failure in the treatment of acid related
diseases has been reported and the degree and speed of onset of
symptom relief are important to patients.sup.21. It has been
estimated that about 30% of GERD patients remain symptomatic on
standard dose of PPI.sup.22. Furthermore PPI's have a short plasma
half life which often leads to nocturnal acid breakthrough.sup.23.
Therapeutic oral doses of PPIs reach steady state and thus achieve
their maximal effective levels after 4-5 days with typical dosing
regimens.sup.24. This slow and cumulative onset of effect of PPIs
relates to their ability to inhibit only those pumps which are
active when the PPI drug is available. After PPI administration,
there is a return of acid secretion that is partly due to de novo
synthesis of the enzyme.sup.25.
[0006] Zinc is an essential part of the diet that all cells require
in order to maintain membrane integrity and function. Deficiency in
intracellular zinc leads to apoptotic events, and cell
death.sup.26-30. Previous studies have investigated the potential
role of zinc in the proliferation and generation of the protective
barrier, namely the mucous gel layer at the surface of the
stomach.sup.31-34. These studies falsely attributed the reduction
in acid secretion to an increase in the thickness of the gel
layer..sup.33-35.
[0007] Gastric acid aids protein digestion; facilitates the
absorption of iron, calcium, and vitamin B12; and prevents
bacterial overgrowth. When levels of acid and proteolytic enzymes
overwhelm the mucosal defense mechanisms, ulcers occur. To avoid
damage that is associated with these harsh conditions, gastric acid
must be finely regulated by overlapping neural (e.g.
acetylcholine), hormonal (e.g. gastrin and ghrelin), and paracrine
(e.g. histamine and somatostatin) pathways, and more recently via
the Calcium Sensing Receptor. Any long term alterations in any of
these regulatory pathways leads to cell and tissue destruction and
clinical manifestations such as peptic ulcer diseases, or
gastroesophageal reflux disease (GERD). Two methods are commonly
employed to treat the overproduction of acid: a) surgically, by
elimination of the neuronal element (vagotomy) or b)
pharmacologically, either through histamine 2 receptor antagonists
or proton pump inhibitors (PPI's) or a combination of both.
[0008] PPI's such as omeprazole are irreversible inhibitors of the
gastric H.sup.+,K.sup.+-ATPase, recently various derivatives of the
parent compound omeprazole that bind to multiple cysteine residues
on the exofacial surface of the H.sup.+,K.sup.+-ATPase have been
developed in hopes of having a tighter molecular binding, and
longer action have been employed. Both rabeprazole, and
lansoprazole are examples of these multiple binding drugs and are
activated in the acidic lumen of the gastric gland and modify the
cysteine residues located on the luminal surface of the
H.sup.+,K.sup.+-ATPase. In the resting cell the acid secreting
pumps are internalized in a system of tubular vesicles, and are in
such a conformational state that the PPIs can only inhibit the
H.sup.+,K.sup.+-ATPases which have already been activated and
transferred to the apical surface of the parietal cell.
[0009] Although optimizing pharmacological profiles within the PPI
class may provide some clinical benefit, other areas of research
may prove to be more fruitful and furthermore the fine tuning of
the acid secretory process is still not completely understood and
remains an important target for therapies to modulate gastric acid
secretion.
[0010] Zinc is required for a large number of biological processes
including gene expression, replication, membrane stability,
hormonal storage and release and as a catalytic component for
enzymes. There has been no investigation of the actions of zinc at
the cellular level relating to effects on acid secretion.
[0011] Helicobacter pylorus resides within the mucous layer of the
human gastric mucosa. Due to extremely low pH, the stomach is a
hostile environment to most other microorganisms. The ability of H.
pylori to flourish in the stomach has been attributed to protective
mechanisms such as its production of urease, protecting the
bacterium from gastric acidity by creating a basic
microenvironment, See. Taylor and Blaser, Epidemiol Rev, 13:42-59,
(1991).
[0012] The stomach is a large organ that can be divided into 3 main
zones that are involved in the process of digestion of foodstuff
and the sterilization of liquids and water. When defining the
functional process of the stomach it has been commonly divided into
two zones: Upper Stomach, and Lower Stomach. The upper stomach, is
thought to be composed of the fundus and upper body, and shows low
frequency, sustained contractions that are responsible for
generating a basal pressure within the stomach. Of note is that
these tonic contractions also generate a pressure gradient from the
stomach to small intestine and are responsible for gastric
emptying. Interestingly, when swallowing food and the consequent
gastric distention that occurs acts to inhibits contraction of this
region of the stomach, allowing it to balloon out forming a large
reservoir without a significant increase in pressure. The lower
stomach is thought to be involved in the grinding and liquefaction
of the foodstuffs by the secretion of HCl from the parietal cells
found in this section of the stomach.
BRIEF DESCRIPTION OF THE FIGURES
[0013] FIG. 1 shows the original tracing of basal acid secretion,
histamine induced acid secretion and inhibition by ZnCl.sub.2.
Single human and rat gastric glands were isolated, loaded with the
pH-sensitive dye BCECF to measure intracellular pH over single
parietal cells and the pH.sub.i recovery rate was calculated from
the slope after an acid load using the NH.sub.4Cl prepulse
technique. (A, C) Intracellular alkalinization stimulated by
histamine (100 .mu.M) in the absence of extracellular Na.sup.+ as a
function of H.sup.+/K.sup.+-ATPase in gastric glands. (B, D)
Histamine induced proton efflux from gastric glands can be blocked
by 300 umol ZnCl.sub.2. (E) Bar graph summarizing data as means SE
(control: n=32 cells, 3 gland, 3 animals; histamine: n=120 cells,
15 glands, 8 animals; histamine+ZnCl.sub.2: n=60 cells, 6 gland, 4
animals).
[0014] FIG. 2 shows that ZnCl.sub.2 inhibits acid secretion in a
dose dependent manner. ZnCl.sub.2 concentration dependence of
H.sup.+/K.sup.+-ATPase activity (intracellular alkalinization
expressed as .DELTA.pH/min) in the presence of 100 .mu.mol
histamine in comparison to basal and histamine induced acid
secretion. (n=40 cells, 3-4 glands, 3-4 animals for each ZnCl.sub.2
concentration).
[0015] FIG. 3 show the fast onset inhibitory effect and
reversibility with ZnCl.sub.2. (A) original tracing shows the fast
inhibitory effect of ZnCl.sub.2 on histamine induced acid
secretion. Histamine (100 .mu.M) was added through the whole
experiment. When the intracellular alkalinization (protonefflux)
was observed, ZnCl.sub.2 (300 .mu.M) was added to the superfusion
bath. The acid secretion was abolished after a few seconds (flat
middle part). After the removal of ZnCl.sub.2 out of the perfusion
bath the drug was washed out and the increase of the intracellular
pH continued. (B) Original tracing shows the reversibility after
the cells where incubated and superfused over 20 min with
ZnCl.sub.2 (300 .mu.M) and histamine (100 .mu.M). After removal of
ZnCl.sub.2 out of the superfusion bath the intracellular
alkalinization (proton extrusion) occur.
[0016] FIG. 4 shows acid secretion after oral ZnCl.sub.2
application. 300 .mu.mol ZnCl.sub.2 was added to the drinking
water. Animals ate and drank as much as control animals. Prior the
experiment they were fasted for 12-18 hours. The histamine induced
acid secretion was measured as described before. The cells of the
ZnCl.sub.2 treated animals showed a lower rate of proton efflux.
150 mg/kg/d: 0.022.+-.0.0045; (n=60 cells, 10 glands, 3 animals),
0.05 mg/kg/d: 0.034.+-.0.0036; (n=60 cells, 6 glands, 4
animals).
[0017] FIG. 5 shows that ZnCl.sub.2 inhibits gastric acid secretion
in freshly isolated rat whole stomach preparation. Ex vivo rat
whole stomach preparations were incubated in HEPES-buffered Ringer
solution (control: n=9), HEPES-buffered Ringer solution plus 100
.mu.M histamine (n=8), or HEPES-buffered Ringer solution plus 100
.mu.mol histamine and 300 .mu.mol ZnCl.sub.2 (n=8). Stomach
preparations incubated with histamine and ZnCl.sub.2 had a higher
pH than those in HEPES-buffered Ringer solution and histamine and
their pH was similar to the pH of the control stomach.
[0018] FIG. 6 shows measurements of whole stomach intraluminal pH
using a number of zinc salts according to the present invention.
Isolated whole stomach preparations from rats were cannulated at
the esophageal and duodenal junction and perfused in vitro with
37.degree. C. pH 7.4 Ringers solution. The blood perfusate was then
exposed to 100 .mu.M Histamine to induce acid secretion. The lumen
of the stomach was infused with 0.5 cc of non-buffered isotonic
saline. In some studies one of the following zinc salts was added
to the lumen perfusate at a final concentration of 300 .mu.M (zinc
chloride, zinc sulfate, zinc acetate, zinc citrate). The data are
the sum of 5 separate stomachs from 5 separate animals for each of
the columns. Data are the mean of all studies with the standard
error of the mean displayed.
[0019] FIG. 7 show the immunohistochemistry in rat stomach fundus.
(A) Immunolocalization of the gastric H.sup.+, K.sup.+-ATPase a
subunit in rat fundic gland parietal cells (40.times.). (B) Fundic
parietal cell Electron Microscopy of Gold Tagged H.sup.+,
K.sup.+-ATPase protein. Here the nucleus, apical membrane and
canaliculus like structure can be seen (8,000.times.). (C) Higher
magnification (25,000.times.) of the same cell. Here the gold
tagged H.sup.+, K.sup.+-ATPase protein can be seen distributed at
the borders of the canaliculus like structure (arrows). (In this
figure: n=nucleus, c=canaliculus like structure, am=apical
membrane)
[0020] FIG. 8 show the original tracing of basal acid secretion and
histamine induced acid secretion in the gastric fundus and corpus.
Single rat gastric glands were isolated, loaded with pH sensitive
dye BCECF to measure intracellular pH of single parietal cells and
the pHi recovery rate was calculated from the slope after an acid
load using the NH.sub.4CL prepulse technique. (A) Original tracing
of an F1 gland alkalinization (proton efflux) after removing
Na.sup.+ out of the perfusion bath. (B) Intracellular
alkalinization of an F1 gland stimulated by histamine (100 .mu.M)
in the absence of extracellular Na.sup.+ as a function of H.sup.+,
K.sup.+-ATPase. (C) Tracing of a corpus gland alkalinization under
resting condition. (D) Intracellular alkalinization of an corpus
gland stimulated by histamine (100 .mu.M) in the absence of
extracellular Na.sup.+ as a function of H.sup.+,
K.sup.+-ATPase.
[0021] FIG. 9 shows a secretagogue series of F1 glands. F1 gland
under basal condition with no stimulation shows alkalinization
rates of 0.039 .DELTA. pH/min 0.009 (n=52 cells/8 glands/5
animals). In the presence of 100 .mu.M histamine recovery rates
were 0.042.+-.0.007 .DELTA. 10 pH.sub.i/min (n=64 cells/8 glands/6
animals). In the presence of 100 .mu.M acetylcholine F1 glands
alkalinized at a rate of 0.075.+-.0.0015 .DELTA. pH.sub.i/min (n=86
cells/10 glands/6 animals). In the presence of 100 .mu.M
pentagastrin F1 glands show alkalinization rates of 0.062.+-.0.007
.DELTA. pH.sub.i/min (n=49 cells/6 glands/5 animals).
[0022] FIG. 10 shows original tracing of acid secretion comparing
F1 glands and Corpus glands with Omeprazole and AZD0865. Single rat
gastric glands were isolated, loaded with the pH sensitive dye
BCECF to measure intracellular pH over single parietal cells and
the pHi recovery rate was calculated from the slope after an acid
load using NH.sub.4Cl prepulse technique as described previously.
(A) Original tracing of an intracellular pH measurement
demonstrating a F1 gland alkalinization after stimulation by
histamine (100 .mu.M). This tracing shows that omeprazole (200
.mu.M) does not inhibit acid secretion in F1 glands. (B) Corpus
gland tracing of intracellular alkalinization after stimulation
with histamine (100 .mu.M). This tracing shows that Omeprazole (200
.mu.molar) inhibits acid secretion in the corpus with a
intracellular alkalinization rate of (0.014.+-.0.002
.DELTA.pH.sub.i/min. (C) Intracellular tracing of pH measurements
demonstrating that AZD0865 does not completely inhibit proton
extrusion in the fundus as it does in the corpus. In fundic glands
which have been exposed to 10 .mu.M of AZD0865 the intracellular
recovery is 0.031.+-.0.006 .DELTA.pH.sub.i/min. (D) In the corpus
AZD0865 shows strong inhibition of potassium dependant recovery
with intracellular alkalinization rates of 0.021.+-.0.008
.DELTA.pH.sub.i/min.
BRIEF DESCRIPTION OF THE INVENTION
[0023] The present invention relates to novel compositions and
methods for the rapid inhibition of acid secretion that has little
to no potential for side effects. In a first aspect, the present
invention relates to zinc compositions comprising at least one
pharmaceutically compatible zinc salt (preferably a water soluble
salt) in an effective amount which produces a rapid decrease (i.e.,
within a period of no greater than about 5 minutes, no greater than
about 10 minutes, no greater than about 20 minutes, no greater than
about 30 minutes, no greater than one hour) of acid secretion in a
patient's stomach with a resulting increase (elevation) in stomach
pH to an intragastric pH level of at least about 3.0-3.5, at least
about 4.0, about 4.0 to about 5.0. In this aspect of the invention,
a patient who is in need of an increase of stomach pH is treated
with an effective amount of a pharmaceutically compatible zinc salt
such that rapid onset of elevated pH within the stomach occurs.
This method invention relies on the administration (preferably by,
but not limited to, ingestion) of an effective amount of at least
one pharmaceutically compatible, preferably water-soluble zinc salt
and in which a substantial portion dissolves in the gastric juices
at low pH (generally less than about 2.0) and preferably within a
range pH from low pH (about 1.0 to about 2.0) to higher pH (about
5.5 to about 7.5 or higher) such that effective amounts of zinc
salt may be administered to provide an initial rapid inhibition of
acid release and a subsequent maintenance of inhibition of acid
release in the stomach. In the present invention, inhibition of
gastric acid is inhibited preferably within a rapid period of about
20 minutes to about 1 hour (generally, within a period no greater
than about 5 minutes, within a period no greater than about 10
minutes or within a period no greater than about 20 minutes, within
a period no greater than about 30 minutes, within a period no
greater than about one hour).
[0024] The rapid decrease of acid secretion in the patient's
stomach occurs throughout the stomach (in both the upper stomach
and lower stomach through inhibition of H.sup.+,K.sup.+-ATPase),
although localized effects of compounds according to the present
invention in the upper stomach, especially in the fundic region of
the stomach (through inhibition of a second distinguishable protein
H.sup.+-ATPase) and/or the upper body of the upper stomach (through
inhibition of H.sup.+,K.sup.+-ATPase). Thus, an additional aspect
of the invention is directed to the use of effective amounts of
pharmaceutically acceptable zinc compounds for the inhibition of
H.sup.+,K.sup.+-ATPase (generally throughout the stomach,
H.sup.+-ATPase (primarily in the fundic region of the stomach) and
preferably both. The finding that the present compounds may be used
to inhibit H.sup.+-ATPase in the fundic region has important
clinical ramifications for the following reasons:
1) The erosion of the esophagus by exposure to acid has life
threatening consequences due to either internal bleeding,
ulceration, and or gastric carcinoid formation by the prolonged
exposure to acid. Pursuant to the present invention, as is now
demonstrated-glands in the fundus are in direct proximity to the
esophageal juncture, that they will secrete acid and can be
inhibited by compounds according to the present invention, thus
making the present compounds particularly effective in treating
GERD, NERD and related conditions. 2) There is an ever increasing
number of patients that are becoming insensitive to PPI (proton
pump inhibitors) and have recurrent symptoms of acid reflux
disease. The protein that we identified in the fundic glands is not
sensitive to PPI's and could be the reason that these patients do
not respond to classical therapy. 3) Patients on PPI's for long
periods of time appear to show some "rebound" acid secretion. This
result could again be linked to the fundic H.sup.+-ATPase, which we
show is sensitive to Histamine and to the levels of protons within
the cell.
[0025] In preferred embodiments of this invention aspect, a single
zinc salt which is water-soluble regardless of pH (i.e., within a
range of pH from about 1.0 to about 7.5 or above) is preferred.
Zinc chloride is the preferred salt for use in the present
invention. In alternative embodiments, a mixture of a low pH
soluble zinc salt with a high pH soluble zinc salt or a zinc salt
which may be readily absorbed through the small intestine (such as
a zinc amino acid chelate compound), optionally in combination with
a pharmaceutically acceptable buffer is provided. In this aspect of
the invention, an effective amount of a zinc salt selected from the
group consisting of zinc chloride (ZnCl.sub.2), zinc acetate, zinc
ascorbate, zinc succinate, zinc tartrate, zinc malate, zinc
maleate, a zinc amino acid chelate (mono- or bis-chelate) and
mixtures thereof, preferably a mixture of zinc chloride and at
least one of zinc acetate, zinc gluconate, zinc succinate, zinc
ascorbate, and a zinc amino acid chelate is provided alone or in
combination with a pharmaceutically acceptable carrier, additive or
excipient.
[0026] In various aspects, the present invention relates to the use
of at least one water-soluble zinc salt alone or in combination
with at least one compound/composition (within the context of the
disease state or condition to be treated) selected from the group
consisting of a traditional proton pump inhibitor
compound/composition, an H2 blocker, an antibiotic/antimicrobial
agent (effective against H. pylori), a cytoprotective agent or a
mixture of these agents (Helidac, Prevpac) to provide fast action
in reducing gastric acid secretion, to lower the pH of the stomach,
to prevent or reduce the likelihood of ulcer disease, to treat
ulcer disease, to treat gastric cancer, to treat a disease or
condition selected from the group consisting of gastroesophageal
reflux disease (GERD), non-erosive reflux disease (NERD),
Zollinger-Ellison syndrome (ZE disease), ulcer disease, and gastric
cancer, as well as preventing or reducing the likelihood of ulcer
disease.
[0027] Pharmaceutical compositions comprising a mixture of zinc
salts which maximize both immediate and extended release
characteristics of the present invention, optionally in combination
with a pharmaceutically acceptable carrier, additive or excipient
and further optionally an effective amount of additional agent
selected from the group consisting of a proton pump inhibitor, an
H2 blocker, an anti-H. pylori antibiotic/antimicrobial, a
cytoprotective agent and a combination of agents, are additional
aspects of the present invention. Any one or more of these
compositions may be used within context to treat the various
conditions/disease states as otherwise disclosed herein.
[0028] In an additional aspect of the present invention,
pharmaceutical compositions comprise at least one water soluble
zinc salt as otherwise described herein in combination with at
least one (additional) therapeutic agent wherein the oral
administration of said agent is favorably affected by elevated pH
levels in the stomach, optionally in combination with a
pharmaceutically acceptable carrier, additive or excipient. It has
unexpectedly been discovered that the inclusion of a water soluble
zinc salt as otherwise described herein to raise the pH of the
gastric juices, in combination with a therapeutic agent which is
favorably responsive to elevated pH levels because of the tendency
of the agent to produce/increase undesired acidity in the stomach,
because of acid sensitivity of the therapeutic agent, because of
enhanced solubility at higher acid pH's (less acid) of about 3.54.0
or higher, and/or because of the tendency of the therapeutic agent
to create GI tract distress or ulcerations at lower pH's, which are
substantially reduced or alleviated at higher pH's represents a
general approach for enhancing the oral administration of
therapeutic agents by increasing bioavailability and/or decreasing
the side effects from the therapeutic agent.
DETAILED DESCRIPTION OF THE INVENTION
[0029] The following terms are used throughout the specification to
describe the present invention.
[0030] The term "patient" or "subject" refers to an animal,
preferably a mammal, even more preferably a human, in need of
treatment or therapy to which compounds according to the present
invention are administered in order to treat a condition or disease
state treatable using compounds according to the present invention.
Depending upon the disease or condition treated the term patient
refers to the animal treated for that disease within context.
[0031] The term "effective" is used to describe a treatment,
compound, composition, component or a related aspect of the present
invention, which, when used in context, produces an intended result
which may include the increase in pH in the stomach, the reduction
of symptoms associated with excess acid release, the enhanced
bioavailability of an administered compound, or the favorable
treatment of a disease state or condition. The term effective
subsumes both an amount or concentration of one or more active
agent(s) as described herein and a period of time which is
consistent with producing an intended effect.
[0032] The term "pharmaceutically acceptable zinc salt" or zinc
salt" used in context, refers to a salt or salt combination which
contains zinc, dissolves in the gastric juices at reduced pH and is
absorbed to some extent in the gastric mucosa at a low pH of about
2 or less, at a higher pH of about 4.0 to 5.0 or above of the
stomach and at the high pH's of the small intestine to reach and
maintain effective concentrations of zinc in the blood stream over
a period of therapy. Exemplary pharmaceutically compatible zinc
salts include both inorganic and organic zinc salts, for example,
zinc acetate, zinc ascorbate, zinc benzoate, zinc bromide, zinc
butyrate, zinc caprylate, zinc carbonate (soluble in dilute acid at
low pH of the stomach), zinc camosine, zinc citrate, zinc chloride,
zinc fluoride, zinc formate, zinc fumarate, zinc fumaric acid
monoethyl ester, zinc gallate, zinc gluconate, zinc glutarate, zinc
glycerate, zinc glycerophosphate, zinc glycolate, zinc hydroxide,
zinc iodide, zinc iodate, zinc lactate, zinc malate, zinc maleate,
zinc myristate, zinc nitrate, zinc oratate, zinc oxide, zinc phenol
sulfonate, zinc phosphate, zinc picolinate, zinc picrate, zinc
propionate, zinc salicylate, zinc selenate, zinc stearate, zinc
succinate, zinc sulfate, zinc tannate, zinc tartrate, zinc
undecylenate, zinc valerate, and zinc chelates, including zinc
amino acid chelates (including, depending on concentration, mono-
and bis-chelates of L- or D-amino acids (preferably, the naturally
occurring L-amino acid which may be more readily absorbed from the
gastrointestinal tract) which complex or chelate with zinc
including preferably. L-arginine (zinc arginate), L-cysteine,
L-cysteine, L-N-acetylcysteine, L-histidine (also D-histidine as
zinc histidinate), L-taurine, L-glycinate, L-aspartate (zinc
aspartate) and L-methionine (zinc methionine), among others. Note
that for purposes of the present invention, zinc chelates,
including zinc nicotinamide complex and zinc amino acid chelates
are considered zinc salts. Preferred zinc salts for use in the
present invention include zinc acetate, zinc arginate, zinc
butyrate, zinc chloride, zinc citrate, zinc formate, zinc fumarate,
zinc gluconate, zinc glutarate, zinc glycerate, zinc glycolate,
zinc histidinate, zinc lactate, zinc malate, zinc maleate, zinc
picolinate, zinc propionate, zinc salicylate, zinc succinate, zinc
sulfate, zinc undecylenate, zinc salt of 1,6 fluctose diphosphate
and mixtures thereof.
[0033] Preferably, the pharmaceutically acceptable zinc salt is
"water soluble". The term "water soluble" is used to describe a
zinc salt (and zinc chelates which fall under this term) according
to the present invention which has a water solubility of at least
about 0.01 moles/Liter, preferably at least about 0.05
moles/Liter.
[0034] One of ordinary skill will recognize favorable zinc salts to
use in the present invention. In aspects of the invention, at least
one pharmaceutically compatible, water-soluble zinc salt is
administered to a patient in order to provide a rapid inhibition of
acid release in the stomach, resulting in an increase in stomach pH
to above 4 (generally between about 4.0 and 5.0, in some cases
above 5.0) for an extended period of time, preferably at least 2
hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 16 hours, 20
hours or more. It is noted that in certain preferred aspects of the
invention, the zinc salt or combination of salts chosen to be
administered to the patient may be adjusted to provide an initial
bolus concentration of zinc in the stomach at low pH in order to
produce the rapid inhibition of acid release and rise in pH in the
stomach to a level above about 4. In addition, a preferred zinc
salt or salt combination inhibits acid release in the stomach at
varying levels of acidity and pH--i.e., at a level which is quit
acidic (pH, less than about 2.0) to a pH of about 4.0 or
higher.
[0035] The term "providing fast action in reducing gastric acid
secretion" is used to describe the fact that the method according
to the present invention results in an increase in pH to a level of
at least about 4.0, more preferably about 4.0 to about 5.0 or
slightly above, in a period of no greater than about 30 minutes,
preferably in less than about 20-30 minutes, even more preferably
in less than about 10-20 minutes, in about 15 minutes or less or
alternatively, in less than about 5 minutes.
[0036] The term "secretagogue" refers to the period during which
time the pariental cells of the stomach secrete acid into the
gastric juices to lower pH. Often the secretagogue period occurs
just after a meal, but the secretion of acid may occur at other
times. The secretagogue phase can be of short duration or longer
duration.
[0037] The term gastroesophageal reflux disease or "GERD" or "acid
reflux" is a condition in which the liquid content of the stomach
regurgitates (backs up, or refluxes) into the esophagus. The liquid
can inflame and damage the lining of the esophagus although this
occurs in a minority of patients. The regurgitated liquid usually
contains acid and pepsin that are produced by the stomach. The
refluxed liquid also may contain bile that has backed-up into the
stomach from the duodenum. Acid is believed to be the most
injurious component of the refluxed liquid. Pepsin and bile also
may injure the esophagus, but their role in the production of
esophageal inflammation and damage (esophagitis) is not as clear as
is the role of acid.
[0038] GERD is a chronic condition. Once it begins, it usually is
life-long. If there is injury to the lining of the esophagus
(esophagitis), this also is a chronic condition. Moreover, after
the esophagus has healed with treatment and treatment is stopped,
the injury will return in most patients within a few months. Once
treatment for GERD is begun, therefore, it may be necessary to
continue the treatment continually, generally for short periods of
time.
[0039] Actually, the reflux of the stomach's liquid contents into
the esophagus occurs in most normal individuals. In fact, one study
found that reflux occurs as frequently in normal individuals as in
patients with GERD. In patients with GERD, however, the refluxed
liquid contains acid more often, and the acid remains in the
esophagus longer.
[0040] Gravity, swallowing, and saliva are important protective
mechanisms for the esophagus, but they are effective only when
individuals are in the upright position. At night while sleeping,
gravity is not in effect, swallowing stops, and the secretion of
saliva is reduced. Therefore, reflux that occurs at night is more
likely to result in acid remaining in the esophagus longer and
causing greater damage to the esophagus.
[0041] Certain conditions make a person susceptible to GERD. For
example, reflux can be a serious problem during pregnancy. The
elevated hormone levels of pregnancy probably cause reflux by
lowering the pressure in the lower esophageal sphincter (see
below). At the same time, the growing fetus increases the pressure
in the abdomen. Both of these effects would be expected to increase
reflux. Also, patients with diseases that weaken the esophageal
muscles (see below), such as scleroderma or mixed connective tissue
diseases, are more prone to develop reflux.
[0042] The cause of GERD is complex. There probably are multiple
causes, and different causes may be operative in different
individuals or even in the same individual at various times. A
number of patients with GERD produce abnormally large amounts of
acid, but this is uncommon and not a contributing factor in the
vast majority of patients. The factors that contribute to causing
GERD are the lower esophageal sphincter, hiatal hernias, esophageal
contractions, and emptying of the stomach. Notwithstanding the
cause of GERD, the present invention may reduce the tendency of
having injurious acid reflux into the esophagus, causing
damage.
[0043] When the wave of contraction in the esophagus is defective,
refluxed acid is not pushed back into the stomach. In patients with
GERD, several abnormalities of contraction have been described. For
example, waves of contraction may not begin after each swallow or
the waves of contraction may die out before they reach the stomach.
Also, the pressure generated by the contractions may be too weak to
push the acid back into the stomach. Such abnormalities of
contraction, which reduce the clearance of acid from the esophagus,
are found frequently in patients with GERD. In fact, they are found
most frequently in those patients with the most severe GERD. The
effects of abnormal esophageal contractions would be expected to be
worse at night when gravity is not helping to return refluxed acid
to the stomach. Note that smoking also substantially reduces the
clearance of acid from the esophagus. This effect continues for at
least 6 hours after the last cigarette.
[0044] Most reflux during the day occurs after meals. This reflux
probably is due to transient LES relaxations that are caused by
distention of the stomach with food. A minority of patients with
GERD, about 20%, has been found to have stomachs that empty
abnormally slowly after a meal. The slower emptying of the stomach
prolongs the distention of the stomach with food after meals.
Therefore, the slower emptying prolongs the period of time during
which reflux is more likely to occur.
[0045] The term "non-erosive reflux disease" or "NERD" is used
describe a specific form of GERD, described above. In some cases,
GERD erodes the esophageal lining, creating a condition called
esophagitis. NERD is GERD that does not cause esophagitis. Because
most GERD sufferers do not have esophagitis, NERD is the most
common form of GERD. Because its name contains the word
"nonerosive," it may appear that NERD is the least severe form of
GERD, but this is not necessarily so. NERD is actually more likely
to produce extra-esophageal complications, and is also less likely
to respond to fundoplication surgery. In one study, only 56% of
NERD patients (compared with 90% of patients with erosive reflux)
reported that their symptoms were completely eliminated with
fundoplication. NERD was also twice as likely to cause swallowing
difficulties.
[0046] Heartburn is the chief symptom of NERD. It has a number of
potential causes, including hiatal hernia, lifestyle behaviors, and
diet. Many people deal with heartburn by simply adjusting their
behavior. In some cases, medication or surgery may be required.
Traditional antacids have also been used to treat NERD.
[0047] The term "Zollinger-Ellison syndrome" or "ZE syndrome" is
used throughout the specification to describe a condition caused by
abnormal production of the hormone gastrin. In ZE syndrome, small
tumor (gastinoma) in the pancreas or small intestine produces the
high levels of gastrin in the blood. ZE syndrome is caused by
tumors usually found in the head of the pancreas and the upper
small bowel. These tumors produce the hormone gastrin and are
called gastrinomas. High levels of gastrin cause overproduction of
stomach acid. High stomach acid levels lead to multiple ulcers in
the stomach and small bowel. Patients with ZE syndrome may
experience abdominal pain and diarrhea. The diagnosis is also
suspected in patients without symptoms who have severe ulceration
of the stomach and small bowel.
[0048] The agents of choice for treating ZE syndrome are the proton
pump inhibitors (PPI) as described hereinabove. These drugs
dramatically reduce acid production by the stomach, and promote
healing of ulcers in the stomach and small bowel. They also provide
relief of abdominal pain and diarrhea.
[0049] Surgical removal of a single gastrinoma may be attempted if
there is no evidence that it has spread to other organs (such as
lymph nodes or the liver). Surgery on the stomach (gastrectomy) to
control acid production is rarely necessary today. Early diagnosis
and surgical removal of the tumor is associated with a cure rate of
only 20% to 25%. However, gastrinomas grow slowly, and patients may
live for many years after the tumor is discovered. Acid-suppressing
medications are very effective at controlling the symptoms of acid
overproduction.
[0050] The term "ulcer" is used throughout the specification to
describe an area of tissue erosion, for example, especially of the
lining of the gastrointestinal (GI) tract, especially of the
stomach (peptic ulcer), esophagus or small intestine (duodenal
ulcer). Due to the erosion, an ulcer is concave. It is always
depressed below the level of the surrounding tissue. Ulcers can
have diverse causes, but in the GI tract, they are believed to be
primarily due to infection with the bacteria H. pyloridus (H.
pylori). GI ulcers, however, may be made worse by stress, smoking
and other noninfectious factors, especially including excessive
stomach acid because a lower pH tends to be a better growth
environment for H. Pyloridus.
[0051] Traditional treatments for H. pyloridus infections include
antimicrobials/antibiotics, such as amoxicillin, clarithromycin
(biaxin), metronidazole (flagyl) and tetracycline ("an anti-H.
pylori agent"); H2-blockers, such as cimetidine (tagamet),
famotidine (pepcid), nizatidine (axid), ranitidine (zantac); proton
pump inhibitors (PPI), such as esomeprazole (nexium), lansoprazole
(prevacid), omeprazole (prilosec), pantoprazole (protonix) and
rabeprazole (aciphex); cytoprotective agents, such as bismuth
subsalicylate, sucralfate; and combination agents, such as Helidac
(bismuth subsalicylate, metronidazole, and tetracycline
combination), Prevpac (lansoprazole, clarithromycin and
amoxicillin).
[0052] The present invention may be used to treat an H. pyloridus
infection in a patient by administering an effective amount of at
least one pharmaceutically acceptable water-soluble zinc salt,
either alone or in combination (preferably, by coadministration)
with at least one other of the traditional treatment modalities, as
described above.
[0053] The term "coadministration" or "combination therapy" is used
to describe a therapy in which at least two active compounds in
effective amounts are used to treat one or more of the disease
states or conditions as otherwise described herein at the same
time. Although the term coadministration preferably includes the
administration of two active compounds to the patient at the same
time, it is not necessary that the compounds be administered to the
patient at the same time, although effective amounts of the
individual compounds will be present in the patient at the same
time. The active compositions may include one or more zinc salts
and/or additional compounds/compositions such as proton pump
inhibitors, H blockers, antibiotics/antimicrobial agents,
cytoprotective agents or combination agents as otherwise described
herein in effective amounts for the disease or condition for which
the compounds are typically used. In addition, coadministration
also contemplates combinations of water soluble zinc salts as
otherwise described herein in combination with at least one
therapeutic agent wherein elevated pH levels provide a favorably
response to the administration of said therapeutic agent.
[0054] The term "favorably responsive to elevated pH levels" or
"favorably orally administered" is used to describe therapeutic
agents which, in orally administered compositions, provide a
favorable response to an elevated pH in the stomach produced by a
water soluble zinc salt according to the present invention, whether
that favorable response is a reduction in gastric irritation from
the therapeutic agent, a reduction in acid generation/production in
the stomach by the therapeutic agent, to increase bioavailability
which is negatively impacted by the sensitivity and/or inactivation
of the therapeutic agent to an acidic environment or because of
increased solubility of the therapeutic agent in gastric juices at
high pH levels. It has unexpectedly been discovered that the
inclusion of a water soluble zinc salt as otherwise described
herein, in combination with a therapeutic agent which is favorably
responsive to elevated pH levels because of the tendency of the
agent to increase acid release and a lowering of pH in the stomach,
because of increased acid sensitivity of the therapeutic agent,
because of enhanced solubility of the agent (with concombinant
increased bioavailability of the therapeutic agent) at higher (less
acid) pH's of about 3.5-4.0 or higher, and/or because of the
tendency of the therapeutic agent to create GI tract distress or
ulcerations at lower pH's, which are substantially reduced or
alleviated at higher pH's results in greater activity and/or fewer
side effects from the therapeutic agent.
[0055] In general, the weight ratio of water soluble zinc salt to
therapeutic agent which is included in combination therapeutic
compositions according to the present invention ranges from about
1:20 to about 20:1, about 1:10 to about 10:1, about 1:5 to about
5:1, about 1:3 to about 3:1, about 1:2 to about 2:1, about 1:1.5 to
about 1.5 to 1, about 1:1. Of course, the weight ratio used in a
particular combination pharmaceutical composition will depend upon
the water solubility of the zinc salt and the activity of the
therapeutic agent in producing a side effect (such as increasing
stomach acid or increasing gastrointestinal distress (GI tract
distress) or the tendency to increase the environment for
ulceration in the gastrointestinal tract), or to be inactivated,
rendered insoluble or have its bioavailability negatively impacted
by stomach acid, etc.
[0056] Therapeutic compounds which may be favorably administered
orally (for the reasons which are outlined above) in combination
with a water soluble zinc salt in the present invention include the
following:
[0057] Chemotherapeutic Agents
[0058] Zn plus chemolitic agents for use in treating intestinal
cancer
[0059] e.g., Cysplatine;
[0060] Zn plus chemolitic agents used to treat whole tissue cancer
with secondary complications in the intestinal track (GI tract
distress) as follows:
[0061] 13-cis-Retinoic Acid;
[0062] 2-CdA (2-Chlorodeoxyadenosine);
[0063] 5-Azacitidine;
[0064] 5-Fluorouracil (5-FU);
[0065] 6-Mercaptopurine (6-MP);
[0066] 6-TG (6-Thioguanine);
[0067] Abraxane;
[0068] Accutane.RTM. (Isotretinoin);
[0069] Actinomycin-D;
[0070] Adriamycin.RTM. (Doxorubicin Hydrochloride);
[0071] Adrucil.RTM. (Fluorouracil);
[0072] Agrylin.RTM. (Anagrelide;
[0073] Ala-Cort.RTM. (Hydrocortisone);
[0074] Aldesleukin;
[0075] Alemtuzumab;
[0076] ALIMTA (Pemetrexed);
[0077] Alitretinoin;
[0078] Alkaban-AQ.RTM. (Vinblastine);
[0079] Alkeran.RTM. (Melphalan);
[0080] All-transretinoic Acid;
[0081] Alpha Interferon;
[0082] Altretamine;
[0083] Amethopterin;
[0084] Amifostine;
[0085] Aminoglutethimide;
[0086] Anagrelide;
[0087] Kidrolase.RTM. (Asparaginase);
[0088] Lanacort.RTM. (Hydrocortone Phosphate);
[0089] L-asparaginase;
[0090] LCR (Leurocristine);
[0091] Lenalidomide;
[0092] Letrozole;
[0093] Leucovorin;
[0094] Leukeran;
[0095] Leukine.TM. (Sargramostim);
[0096] Leuprolide;
[0097] Leurocristine;
[0098] Leustatin.TM. (Cladribin);
[0099] Liposomal Ara-C;
[0100] Liquid Pred.RTM. (Deltasone);
[0101] Lomustine;
[0102] L-PAM (L-phenylalanine mustard, phenylalanine mustard);
[0103] L-Sarcolysin;
[0104] Lupron.RTM. (Leuprolide Acetate Inj);
[0105] Lupron Depot.RTM. (Leuploride Acetate);
[0106] Matulane.RTM. (Procarbazine);
[0107] Maxidex;
[0108] Mechlorethamine;
[0109] Mechlorethamine Hydrochloride;
[0110] Medralone.RTM. (Methylprednisolone);
[0111] Medrol.RTM. (Methylprednisolone);
[0112] Megace.RTM. (Megestrol Acetate);
[0113] Megestrol;
[0114] Megestrol Acetate;
[0115] Melphalan;
[0116] Mercaptopurine;
[0117] Mesna;
[0118] Mesnex.TM. (Mesna);
[0119] Methotrexate;
[0120] Anandron.RTM. (Nilutamide);
[0121] Anastrozole
[0122] Arabinosylcytosine;
[0123] Ara-C;
[0124] Aranesp.RTM. (Darbepoetin Alfa);
[0125] Aredia.RTM. (Pamidronate);
[0126] Arimidex.RTM. (Anastrozole);
[0127] Aromasin.RTM. (Exemestane);
[0128] Arranon.RTM. (Nelarabine);
[0129] Arsenic Trioxide;
[0130] Asparaginase;
[0131] ATRA (Atragen);
[0132] Avastin.RTM. (Bevacizumab);
[0133] Azacitidine;
[0134] BCG (Bacillus Calmette Guerin);
[0135] BCNU (Carmustine);
[0136] Bevacizumab;
[0137] Bexarotene;
[0138] BEXXAR.RTM. (Tositumomab and Iodine 1131 Tositumomab);
[0139] Bicalutamide;
[0140] BiCNU (CARMUSTINE);
[0141] Blenoxane.RTM. (Bleomycin Sulfate);
[0142] Bleomycin;
[0143] Bortezomib;
[0144] Busulfan;
[0145] Busulfex (Busuflan);
[0146] C225 (Eribitux);
[0147] Calcium Leucovorin;
[0148] Campath.RTM. (Alemtuzumab;
[0149] Camptosar.RTM. (Irinotecan hydrochloride);
[0150] Camptothecin-11;
[0151] Capecitabine;
[0152] Carac.TM. (Fluorouracil);
[0153] Carboplatin;
[0154] Carmustine;
[0155] Carmustine Wafer
[0156] Casodex.RTM. (Bicalutamide);
[0157] CC-5013 (Revlimid);
[0158] CCNU (lomustine);
[0159] CDDP (Cisplatin);
[0160] CeeNU;
[0161] Cerubidine 1 (Daunorubicin);
[0162] Cetuximab;
[0163] Chlorambucil;
[0164] Cisplatin;
[0165] Methotrexate Sodium;
[0166] Methylprednisolone;
[0167] Meticorten.RTM. (prednisone);
[0168] Mitomycin;
[0169] Mitomycin-C;
[0170] Mitoxantrone;
[0171] M-Prednisol.RTM. (Methlyprednisolone);
[0172] MTC (Mitomycin);
[0173] MTX (Methotrexate);
[0174] Mustargen.RTM. (Mechlorethamine HCl);
[0175] Mustine;
[0176] Mutamycin.RTM. (Mitomycin);
[0177] Myleran.RTM. (Busulfan);
[0178] Mylocel.TM. (Hydroxyurea);
[0179] Mylotarg.RTM. (Gemtuzumab Ozogamicin);
[0180] Navelbine.RTM. (Vinorelbine Tartrate);
[0181] Nelarabine
[0182] Neosar.RTM. (Cyclophosphamide);
[0183] Neulasta.RTM. (Pegfilgrastim);
[0184] Neumega.RTM. (Oprelvekin);
[0185] Neupogen.RTM. (Filgrastim);
[0186] Nexavar.RTM. (Sorafenib);
[0187] Nilandron.RTM. (Nilutamide);
[0188] Nilutamide;
[0189] Nipent.RTM. (Pentostatin);
[0190] Nitrogen Mustard;
[0191] Novaldex.RTM. (Genox);
[0192] Novantrone.RTM. (Mitoxantrone);
[0193] Octreotide;
[0194] Octreotide acetate;
[0195] Oncospar.RTM. (Pegylated asparaginase);
[0196] Oncovin.RTM. (Vincristine Sulfate);
[0197] Ontak.RTM. (Denileukin Diftitox);
[0198] Onxal.TM. (Paclitaxel);
[0199] Oprevelkin;
[0200] Orapred.RTM. (Prednisolone Sodium Phosphate);
[0201] Orasone.RTM. (prednisone);
[0202] Oxaliplatin;
[0203] Paclitaxel;
[0204] Paclitaxel Protein-bound;
[0205] Pamidronate;
[0206] Panitumumab;
[0207] Panretin.RTM. (Alitretinoin);
[0208] Paraplatin.RTM. (Paraplatin);
[0209] Citrovorum Factor;
[0210] Cladribine;
[0211] Cortisone;
[0212] Cosmegen.RTM. (Dactinomycin);
[0213] CPT-11 (Topotecan);
[0214] Cyclophosphamide;
[0215] Cytadren @(Aminoglutethimide);
[0216] Cytarabine;
[0217] Cytarabine Liposomal;
[0218] Cytosar-U.RTM. (Cytarabine);
[0219] Cytoxan.RTM. (Cyclophosphamide);
[0220] Dacarbazine;
[0221] Dacogen;
[0222] Dactinomycin;
[0223] Darbepoetin Alfa;
[0224] Dasatinib;
[0225] Daunomycin;
[0226] Daunorubicin;
[0227] Daunorubicin Hydrochloride;
[0228] Daunorubicin Liposomal;
[0229] DaunoXome.RTM. (Daunorubicin Liposoma);
[0230] Decadron;
[0231] Decitabine;
[0232] Delta-Cortef.RTM. (Prednisolone);
[0233] Deltasone.RTM. (Prednisone);
[0234] Denileukin diftitox;
[0235] DepoCyt.TM. (Cytarabine liposome injection);
[0236] Dexamethasone;
[0237] Dexamethasone acetate;
[0238] Dexamethasone Sodium Phosphate;
[0239] Dexasone;
[0240] Dexrazoxane;
[0241] DHAD (Novantrone);
[0242] DIC (Disseminated intravascular coagulation);
[0243] Diodex;
[0244] Docetaxel;
[0245] Doxil.RTM. (Doxorubicin HCl liposome);
[0246] Doxorubicin;
[0247] Doxorubicin liposomal;
[0248] Droxia.TM. (Hydroxyurea);
[0249] DTIC (Dacarbazine);
[0250] DTIC-Dome.RTM. (dacarbazine);
[0251] Duralone.RTM.;
[0252] Efudex.RTM. (fluorouracil topical);
[0253] Eligard.TM. (Leuprolide Acetate);
[0254] Pediapred.RTM. (Prednisolone Sodium);
[0255] PEG Interferon;
[0256] Pegaspargase;
[0257] Pegfilgrastim;
[0258] PEG-INTRON.TM. (Peginterferon alfa-2b);
[0259] PEG-L-asparaginase;
[0260] PEMETREXED;
[0261] Pentostatin;
[0262] Phenylalanine Mustard;
[0263] Platinol.RTM. (Cisplatin);
[0264] Platinol-AQ.RTM. (Cisplatin);
[0265] Prednisolone;
[0266] Prednisone;
[0267] Prelone.RTM. (Prednisolone);
[0268] Procarbazine;
[0269] PROCRIT.RTM. (Epoetin Alfa);
[0270] Proleukin.RTM. (Aldesleukin);
[0271] Prolifeprospan 20 with Carmustine;
[0272] Implant;
[0273] Purinethol.RTM. (Mercaptopurine);
[0274] Raloxifene;
[0275] Revlimid.RTM. (Lenalidomide);
[0276] Rheumatrex.RTM. (Trexall);
[0277] Rituxan.RTM. (Rituximab);
[0278] Rituximab;
[0279] Roferon-A.RTM. (Interferon Alfa-2a);
[0280] Rubex.RTM. (adriamycin)
[0281] Rubidomycin hydrochloride;
[0282] Sandostatin.RTM. (Octreotide Acetate);
[0283] Sandostatin LAR.RTM. (Octreotide Acetate inj);
[0284] Sargramostim;
[0285] Solu-Cortef.RTM. (Hydrocortisone Sodium Succinate);
[0286] Solu-Medrol.RTM. (Methylprednisolone sodium succinate);
[0287] Sorafenib;
[0288] SPRYCEL.TM. (Dasatinib);
[0289] STI-571 (Gleevec);
[0290] Streptozocin;
[0291] SUl1248;
[0292] Sunitinib;
[0293] Sutent.RTM. (Sunitinib Malate);
[0294] Tamoxifen;
[0295] Tarceva.RTM. (Erlotinib);
[0296] Targretin.RTM. (Bexarotene);
[0297] Taxol.RTM. (Paclitaxel);
[0298] Ellence.TM. (Epirubicin hydrochloride);
[0299] Eloxatin.TM. (Oxaliplatin Inj);
[0300] Elspar.RTM. (Asparaginase);
[0301] Emcvtg @(Estramustine);
[0302] Epirubicin;
[0303] Epoetin alfa;
[0304] Erbitux.TM. (Cetuximab);
[0305] Erlotinib;
[0306] Erwinia L-asparaginase;
[0307] Estramustine;
[0308] Ethyol;
[0309] Etopophos.RTM. (Etoposide Phosphate);
[0310] Etoposide;
[0311] Etoposide Phosphate;
[0312] Eulexin.RTM. (Flutamide);
[0313] Evista.RTM. (Raloxifene);
[0314] Exemestane;
[0315] Fareston.RTM. (Toremifene);
[0316] Faslodex.RTM. (Fulvestrant);
[0317] Femara.RTM. (Letrozole);
[0318] Filgrastim;
[0319] Floxuridine;
[0320] Fludara.RTM. (Fludarabine);
[0321] Fludarabine;
[0322] Fluoroplex.RTM. (Fluorouracil topical);
[0323] Fluorouracil;
[0324] Fluorouracil (cream);
[0325] Fluoxymesterone;
[0326] Flutamide;
[0327] Folinic Acid;
[0328] FUDR.RTM. (Floxuridine);
[0329] Fulvestrant;
[0330] G-CSF (Neupogen);
[0331] Gefitinib;
[0332] Gemcitabine;
[0333] Gemtuzumab ozogamicin;
[0334] Gemzar.RTM. (Gemcitabine);
[0335] Gleevec.TM.;
[0336] Gliadel.RTM. (Carmustine Wafer);
[0337] GM-CSF;
[0338] Goserelin;
[0339] Granulocyte--Colony Stimulating Factor;
[0340] Granulocyte Macrophage Colony Stimulating Factor;
[0341] Taxotere.RTM. (Docetaxel);
[0342] Temodar.RTM. (Temozolomide);
[0343] Temozolomide;
[0344] Teniposide;
[0345] TESPA (Thiotepa);
[0346] Thalidomide;
[0347] Thalomid.RTM. (Thalidomide);
[0348] TheraCys.RTM. (Intravesical);
[0349] Thioguanine;
[0350] Thioguanine Tabloid @(Thioguanine);
[0351] Thiophosphoamide;
[0352] Thioplex.RTM. (Thiotepa);
[0353] Thiotepa;
[0354] TICE.RTM. (Bacillus of Calmette and Guerin);
[0355] Toposar.RTM. (Etoposide);
[0356] Topotecan;
[0357] Toremifene;
[0358] Tositumomab;
[0359] Trastuzumab;
[0360] Tretinoin;
[0361] Trexall.TM. (Methotrexate);
[0362] Trisenox.RTM. (Arsenic);
[0363] TSPA (Thiotepa);
[0364] VCR;
[0365] Vectibix.TM. (Panitumumab);
[0366] Velban.RTM. (Vinblastine Sulfate);
[0367] Velcade.RTM. (Bortezomib);
[0368] VePesid.RTM. (Etoposide);
[0369] Vesanoid @(Tretinoin);
[0370] Viadur.TM. (Leuprolide Acetate Implant);
[0371] Vidaza.RTM. (Azacitidine);
[0372] Vinblastine;
[0373] Vinblastine Sulfate;
[0374] Vincasar Pfs.RTM. (Vincristine Sulfate Injection);
[0375] Vincristine;
[0376] Vinorelbine;
[0377] Vinorelbine tartrate;
[0378] VLB (Vinblastine Sulfate);
[0379] VM-26;
[0380] Vorinostat;
[0381] VP-16 (Etoposide);
[0382] Vumon.RTM. (Teniposide);
[0383] Xeloda.RTM. (Capecitabine);
[0384] Zanosar.RTM. (Streptozocin);
[0385] Halotestin.RTM. (Fluoxymesterone);
[0386] Herceptin.RTM. (Trastuzumab);
[0387] Hexadrol;
[0388] Hexalen.RTM. (Altretamin);
[0389] Hexamethylmelamine;
[0390] HMM (antineoplastic or cytotoxic);
[0391] Hycamtin.RTM. (Hycamtin);
[0392] Hydrea.RTM. (Hydroxyurea);
[0393] Hydrocort Acetate.RTM. (Hydrocortisone);
[0394] Hydrocortisone;
[0395] Hydrocortisone Sodium Phosphate;
[0396] Hydrocortisone Sodium Succinate;
[0397] Hydrocortone Phosphate;
[0398] Hydroxyurea;
[0399] Ibritumomab;
[0400] Ibritumomab Tiuxetan;
[0401] Idamycin.RTM. (Idarubicin);
[0402] Idarubicin;
[0403] Ifex.RTM. (Ifosfamide);
[0404] IFN-alpha;
[0405] Ifosfamide;
[0406] IL-11;
[0407] IL-2;
[0408] Imatinib mesylate;
[0409] Imidazole Carboxamide;
[0410] Interferon alfa;
[0411] Interferon Alfa-2b (PEG Conjugate);
[0412] Interleukin-2;
[0413] Interleukin-11;
[0414] Intron A.RTM. (interferon alfa-2b);
[0415] Iressa.RTM. (Getfitinib);
[0416] Irinotecan;
[0417] Isotretinoin;
[0418] Zevalin.TM.;
[0419] Zinecard.RTM. (Dexrazoxane);
[0420] Zoladex.RTM. (Goserelin);
[0421] Zoledronic acid;
[0422] Zolinza;
[0423] Zometa.RTM. (Zoledronic Acid for Inj).
[0424] Immunosuppression Agents
[0425] Zn plus agents used as immunosuppressive agents following
organ transplantation such as cyclosporine and its derivatives,
azathioprine, 6-mercaptopurine Prednisone, infliximab (Remicade),
and tetracycline, among others.
[0426] Assorted Medications
[0427] Zn given in combination with asthma related drugs:
Theophylline and cortisteroids, including betamethasone, cortisone,
dexamethasone, hydrocortisone, methylprednisolone, prednisolone and
budenoise. Each of these agents can induce stomach lining erosion
and increased acid secretion.
[0428] NSAIDS:
[0429] Over the counter NSAIDS and associated compounds cause GERD
and GERD symptoms including ulcer disease:
TABLE-US-00001 OTC Name Generic Name Actron .RTM. ketoprofen Advil
.RTM. ibuprofen Aleve .RTM. naproxen sodium Bayer .RTM. aspirin
Ecotrin .RTM. aspirin Excedrin .RTM. aspirin, acetaminophen and
caffeine Motrin IB .RTM. ibuprofen Nuprin .RTM. ibuprofen Orudis KT
.RTM. ketoprofen
[0430] Antidepressive Agents
[0431] Selective serotonin reuptake inhibitors (SSRIs): Citalopram
(Celexa), Escitalopram (Lexapro), Fluoxetine (Prozac); Parozxetine
(Paxel) and Sertraline (Zoloft.)
[0432] Zn taken in combination with the following drugs could
potentially increase bioavailability of molecule due to the fact
zinc does not cause the effects on inhibiting Cytochromes P450 2C9
& 3A4 as omeprazole or esoprazole. [0433] Carbamazepine; [0434]
Cyclosporine; [0435] Diazepam, other benzodiazepines; [0436]
Diltiazem, Nifedipine, Verapamil; [0437] Erythromycin,
Clarithromycin; [0438] Lidocaine; [0439] Lovastatin, other statins
including Atorvastatin, [0440] Phenytoin; [0441] Quinidine; [0442]
Terfenadine
[0443] The term "proton pump inhibitor" is used throughout the
specification to describe Proton pump inhibitors as drugs that help
control the painful discomfort of heartburn and gastroesophageal
reflux disease (GERD), and promote the healing of stomach and
duodenal ulcers. Proton pump inhibitors are only available by
prescription. They come as tablets, capsules, injections, or
powders that are made into a suspension.
[0444] Proton inhibitors work by blocking the production of stomach
acid. They inhibit a system in the stomach known as the proton
pump, which is another name for the "hydrogen-potassium adenosine
triphosphate enzyme" system. Proton pump inhibitors are rather
versatile. They are used to heal stomach and duodenal ulcers,
including stomach ulcers caused by taking nonsteroidal
anti-inflammatory drugs. They are also used to relieve symptoms of
oesophagitis (inflammation of the oesophagus or gullet) and severe
gastroesophageal reflux (GERD) as discussed above.
[0445] Combined with certain antibiotics (such as amoxycillin and
clarithromycin) or with zinc salts according to the present
invention, proton pump inhibitors are effective for treating
Helicobacter pylori infections (a bacterial infection of the
stomach). The H. pylori bacterium is a chief suspect in the cause
of recurring stomach ulcers. PPIs are also a first-choice treatment
for the rare condition called Zollinger-Ellison syndrome, discussed
above.
[0446] Proton Pump Inhibitors exhibit side effects, although they
tend to be manageable, including diarrhea, feeling or being sick,
constipation, flatulence, abdominal pain, headaches and more
rarely, allergic reactions, itching, dizziness, swollen ankles,
muscle and joint pain, blurred vision, depression and dry mouth,
among others. Long-term use of proton pump inhibitors can result in
stomach infections. Because proton pump inhibitors completely stop
acid production--and stomach acid helps kill microbes such as
bacteria in the stomach--using PPIs can lead to growth of
potentially harmful microbes in the stomach.
[0447] Proton pump inhibitors exhibit significant, sometimes
deleterious drug interactions, including reactions with phenytoin
as an epilepsy agent and warfarin to prevent blood clots, to
increase their effects, with ketoconazole and itraconazole to
reduce their absorptivity, with diazepam (valium) to decrease its
metabolism.
[0448] Proton pump inhibitors are usually taken for 1-2 months, but
in some cases may be taken longer. Symptoms may return when a
person stops taking a proton pump inhibitor. Proton pump inhibitors
may cause internal bleeding, signs of which include vomiting blood,
detecting a substance-like coffee grounds in your vomit, or pass
black tarry stools, see your doctor immediately.
[0449] Common proton pump inhibitors include omeprazole (Prilosec),
esomeprazole (Nexium), lansoprazole (Prevacid), pantoprazole
(Protonix) and rabeprazole sodium (Aciphex).
[0450] The present invention relates to a method for providing fast
action with optional long duration effect in reducing gastric acid
secretion, raising the pH of the stomach during a resting phase,
decreasing the duration of stomach acid release during a
secretagogue phase and for treating conditions including
gastroesophageal reflux disease (GERD), non-erosive reflux disease
(NERD), Zollinger-Ellison syndrome (ZE disease), ulcer disease, and
gastric cancer where the reduction in gastric acid secretion is
beneficial, as well as preventing or reducing the likelihood of
ulcer disease by reducing gastric acid section. In addition, the
present methods are useful for treating patients who are
non-responsive to proton pump inhibitors (PPI) and as an
alternative to traditional therapies or conditions which are caused
by rapid and complete inhibition of secretagogue induced acid
secretion.
[0451] The method comprises administering an effective amount of at
least one pharmaceutically acceptable water-soluble zinc salt to
alleviate or treat the condition or disease state. The methods may
involve the administration of a water-soluble zinc salt alone or in
combination with other agents as disclosed herein a single time, or
preferably for longer duration, usually about 2-3 days to about 2-3
months, with varying intervals in between, depending upon the
prognosis and outcome of the treatment.
[0452] Zinc salts according to the present invention may be
administered alone or in combination with other compounds,
compositions or therapies, depending upon the condition or disease
state to be treated, including an effective amount of a proton pump
inhibitor or other agent as otherwise described herein which may be
used to treat H. pylori infections. These agents include proton
pump inhibitors such as esomeprazole, lansoprazole, omeprazole,
pantoprazole or rabeprazole, H2 blockers such as cimetidine,
famotidine, nizatidine or ranitidine, anti-H. pylori agents, such
amoxicillin, clarithromycin (biaxin), metronidazole (flagyl) or
tetracycline, cytoprotective agents such as bismuth subsalicylate
or sucralfate, or a combination agent such as Helidac or
Prevpac.
[0453] In a preferred aspect of the invention, at least one
water-soluble zinc salt is used wherein the zinc salt or
combination is characterized as being soluble and absorbable
(through the gastrointestinal mucosa) at both low pH (i.e., a pH of
about 1-2, which occurs in an acidic condition in the stomach) and
higher pH (i.e., a pH of about 4-5 or slightly above after acid
secretion in the stomach is inhibited or even higher--i.e., a pH of
about 5.5-6.0 in the duodenum to about 6.5-7.5 in the jejunum and
ileum--the pH is slightly higher in the ileum than in the jejunum).
By providing for compositions which are both water-soluble and
absorbable throughout the gastrointestinal mucosa (i.e. in the
stomach and through the various sections of the small intestine),
the bioavailability of the zinc salt will be maximized as will
favorable therapy of the conditions or disease states to be
treated. In this aspect, a preferred combination of effective
amounts of zinc chloride and at least one zinc salt preferably
selected from the group consisting of zinc acetate, zinc arginate,
zinc butyrate, zinc citrate, zinc formate, zinc fumarate, zinc
gluconate, zinc glutarate, zinc glycerate, zinc glycolate, zinc
histidinate, zinc lactate, zinc malate, zinc maleate, zinc
picolinate, zinc propionate, zinc salicylate, zinc succinate, zinc
sulfate, zinc undecylenate, zinc salt of 1,6 fluctose diphosphate
and mixtures thereof, more preferably, zinc acetate, zinc
gluconate, zinc ascorbate, zincx succinate and a zinc amino acid
chelate (as mono- or bis-amino acid chelate) is preferred, although
numerous other zinc acid compounds may be combined to produce
favorable results.
[0454] Preferred zinc salts include those salts in which the
anionic counterion in protonated form has a pKa of at least about 4
to about 5.5 or higher. Mixtures of zinc salts wherein all of the
zinc salts are soluble within a range of pH from 1-2 to about 7.5
are preferred. Zinc acetate, zinc gluconate, zinc glycolate, zinc
succinate and zinc ascorbate alone or in combination with another
zinc salt, especially zinc chloride, are particularly useful for
use in the present invention. Zinc chelates, especially zinc amino
acid chelates (mono- or bis-amino acid chelates) may also be
preferably used wherein a combination of zinc chloride and a zinc
amino acid chelate selected from the group consisting of zinc
chelates (mono- or bis-chelates) of L-cysteine, L-cystine,
L-N-acetylcysteine, L-histidine, D-histidine, L-taurine,
L-glycinate, L-aspartate, L-methionine, and mixtures thereof.
[0455] Note that the following zinc salts have solubilities which
tend to be reduced at pH values above about 7.0, so approaches to
formulation should accommodate such information where absorptivity
from the small intestine is featured (duodenum, jejunum and ileum),
especially at the distil end (jejunum, ileum) where the pH of the
small intestine may rise to between 7-8.0. Note that the use of
such salts may favorably influence release characteristics of
formulations and provide a means of delivering therapeutic agents,
especially those which are administered in combination therapy
according to the present invention. Zinc salts that become
insoluble above a pH of 7 include zinc acetate, zinc chloride, zinc
bromide, zinc fluoride, zinc iodide, zinc sulfate, zinc citrate,
zinc lactate, zinc nitrate, zinc propionate, zinc salicylate, zinc
tartrate, zinc valerate, zinc gluconate, zinc selenate, zinc
benzoate, zinc formate, zinc glycerophosphate, zinc picrate, zinc
butyrate, and the like, and combinations thereof.
[0456] Preferred zinc salts according to the invention include zinc
chloride (where pKa of the counterion is not important because of
its interaction with chloride channels) and organic acids including
zinc acetate (pka 4.75), zinc gluconate, zinc succinate, zinc
tartrate, zinc malate, zinc maleate, zinc zinc ascorbate (pka of
4.2 and 11.6). Other zinc salts of organic acids may also be
preferred, depending on context of use. In addition, zinc glycolate
and zinc lactate may also be used preferably, zinc glycolate being
preferred. Other preferred salts include, for example, zinc
acetate, zinc arginate, zinc butyrate, zinc chloride, zinc citrate,
zinc formate, zinc fumarate, zinc gluconate, zinc glutarate, zinc
glycerate, zinc glycolate, zinc histidinate, zinc lactate, zinc
malate, zinc maleate, zinc picolinate, zinc propionate, zinc
salicylate, zinc succinate, zinc sulfate, zinc undecylenate, zinc
salt of 1,6 fluctose diphosphate and mixtures thereof. In aspects
of the invention, it is preferred that when a combination of zinc
salts is used that at least one zinc salt which is effective at low
pH in the stomach (for immediate inhibition of acid secretion) be
combined with an agent which may exhibit a heightened effect in the
stomach at a pH of 4.0-5.0 or higher, or which is preferentially
absorbed in the small intestine (a zinc mono- or bis-amino acid
chelate or other chelate).
[0457] While not being limited by way of theory, it is believed
that a combination of a zinc salt which is effective at low pH in
the stomach (such as zinc chloride and also zinc sulfate) and one
or more of the organic acid zinc salts as otherwise disclosed
herein which are effective at a higher pH, will maximize delivery
of zinc to the stomach mucosa to obtain a favorable effect, at
first by being dissolved in acid gastric juice in the stomach where
an initial inhibition of acid occurs and the pH rises, and
subsequently, through absorption of zinc (from a zinc salt) at a
higher pH in the stomach or in the small intestine where blood
levels of zinc will increase to therapeutic levels. The absorption
and effect of a zinc salt at higher pH levels in the stomach or at
the higher pH of the small intestine (5.5-7.5 or higher) is
advantageous because this delayed absorption of zinc will reduce
gastric acid secretion at a later time (than an initial effect at a
low pH) over an extended period of time. Compositions according to
the present invention may be administered a single time, but
usually are administered preferably once or twice daily orally for
a period ranging from about 2-3 days to several months or
longer.
[0458] Compositions according to the present invention also relate
to sustained or extended release formulations which comprise a
first component which allows or facilitates fast dissolution in the
gastric juices at low pH so that a rapid inhibition of acid
secretion is effected (with concombinant increase in pH to a level
of about 4.0 to about 5.0 or higher) and a second component which
releases zinc salt at the higher pH level in the stomach or more
preferably, further in the small intestine on a sustained release
basis in order to maintain an effective level of zinc in the blood
stream to inhibit gastric acid secretion in the stomach for
extended periods. The first fast-acting component may be readily
formulated using a zinc salt which dissolves in gastric juice at
low pH (e.g., zinc chloride or zinc sulfate at a pH about 1.0 to
about 2.0) using standard excipients such as lactose,
confectioner's sugar in powered form, various stearate salts, etc,
which dissolves rapidly in the stomach and a second sustained or
extended release formulation which makes us of any number of
polymeric binders, matrices (polymeric and/or erodible), granules,
or enteric coatings to allow release of zinc salt on an extended or
sustained release basis in the small intestine. Many of these
techniques are well known in the art. Exemplary patents such as
U.S. Pat. No. 4,863,741 to Becker, U.S. Pat. No. 4,938,967 to
Newton, et al., U.S. Pat. No. 4,940,556 to MacFarlane, et al., and
U.S. Pat. No. 5,202,128 to Morella, et al., among numerous others,
may be useful for providing teachings, all well known in the art,
for formulating fast release/sustained or extended release
formulations useful in the present invention.
[0459] The above formulations may be useful for providing enhanced
bioavailability of one or more zinc salts and optionally, other
agents which may be useful in treating or reducing the likelihood
of one or more of gastric ulcers, GERD, NERD, Zollinger-Ellison
syndrome, gastric cancer and reducing/inhibiting the secretion of
acid in the stomach and raising the pH of the stomach to about 4.0
to about 5.0 or more, as otherwise disclosed in the present
invention. It is noted that in inhibiting acid secretion in the
stomach, blood concentrations of zinc salt of about 100 micromolar
(.mu.mol) produce inhibition of about 70%. With 300 .mu.mol
concentration of zinc salt, the inhibition approaches 100%. The
time of action of inhibition from the blood delivery side at 100
.mu.mol or 300 .mu.mol is immediate (i.e., as soon as the zinc salt
comes into contact with the cell membrane, inhibition occurs. It
may be shown that inhibition occurs within about 10-15 minutes to 1
about hour in the presence of secretagogue. The zinc salts may be
administered orally (preferably no more than once or twice a day)
or intravenously, alone or in combination with optional PPI
drugs.
[0460] The use of zinc chloride alone, or in combination with at
least one additional zinc salt as otherwise described herein is
preferred. Additional preferred zinc salts include zinc acetate,
zinc gluconate, zinc ascorbate, zinc succinate, and zinc amino acid
chelates (mono- and bis-amino acid chelates). These zinc salts and
combinations may be used alone or in combination with additional
agents such as a proton pump inhibitor (esomeprazole, lansoprazole,
omeprazole, pantoprazole or rabeprazole), an H2 blocker
(cimetidine, famotidine, nizatidine or ranitidine), an anti-H.
pylori agent (amoxicillin, clarithromycin, metronidazole or
tetracycline), a cytoprotective agent such as bismuth subsalicylate
or sucralfate, or a combination agent such as Helidac or
Prevpac.
[0461] Pharmaceutical compositions comprising an effective amount
of a pharmaceutically acceptable zinc salt alone, or preferably in
combination with at least one other zinc salt or an effective
amount of a traditional proton pump inhibitor such as such as
esomeprazole, lansoprazole, omeprazole, pantoprazole or
rabeprazole, an H2 blocker such as cimetidine, famotidine,
nizatidine or ranitidine, an anti-H. pylori agent, such
amoxicillin, clarithromycin (biaxin), metronidazole (flagyl) or
tetracycline, a cytoprotective agent such as bismuth subsalicylate
or sucralfate, or a combination agent such as Helidac or Prevpac,
optionally in combination with a pharmaceutically acceptable
carrier, additive or excipient.
[0462] Pharmaceutical formulations include those suitable for oral,
rectal, nasal, topical (including buccal and sub-lingual), vaginal
or parenteral (including intramuscular, sub-cutaneous and
intravenous) administration. Oral compositions or parenteral
compositions (especially those for IV administration) are
preferred. Compositions according to the present invention may also
be presented as a bolus, electuary or paste. Tablets and capsules
for oral administration may contain conventional excipients such as
binding agents, fillers, lubricants, disintegrants, or wetting
agents. The tablets may be coated according to methods well known
in the art. Oral liquid preparations may be in the form of, for
example, aqueous or oily suspensions, solutions, emulsions, syrups
or elixirs, or may be presented as a dry product for constitution
with water or other suitable vehicle before use. Such liquid
preparations may contain conventional additives such as suspending
agents, emulsifying agents, non-aqueous vehicles (which may include
edible oils), or preservatives. When desired as discussed
hereinabove, the present formulations may be adapted to provide
sustained release characteristics of the active ingredient(s) in
the composition using standard methods well-known in the art. A
composition which provides an effective amount of initial dose of
zinc salt in the gastric juice at low pH followed by extended
release effects of zinc over a longer duration may be
preferred.
[0463] In the case of combination pharmaceutical compositions,
i.e., a composition which comprises at least one water soluble salt
in combination with a therapeutic agent (i.e., other than the zinc
salt), compositions may be formulated in admixture or they may be
compartmentalized in the dosage form. Pharmaceutical formulations
may be formulated in admixture by mixing the actives together along
with the pharmaceutically acceptable carriers, additives and/or
excipients in powder or liquid form and then using directly or
presenting the mixture in tablet or capsule form. The compositions
may be immediate release, sustained or controlled release or
intermediate sustained or controlled release, depending upon the
results desired. Formulations may also be presented which
compartmentalize the water soluble zinc salt and the therapeutic
agent into more than one portion of a tablet or capsule to take
advantage of differential solubilities in order to enhance the
bioavailability of the zinc salt and/or the additional therapeutic
agents, using methods which are readily available in the art to
those of ordinary skill.
[0464] In the pharmaceutical aspect according to the present
invention, the compound(s) according to the present invention is
formulated preferably in admixture with a pharmaceutically
acceptable carrier. In general, it is preferable to administer the
pharmaceutical composition orally, but certain formulations may be
preferably administered parenterally and in particular, in
intravenous or intramuscular dosage form, as well as via other
parenteral routes, such as transdermal, buccal, subcutaneous,
suppository or other route, including via inhalation intranasally.
Oral dosage forms are preferably administered in tablet or capsule
(preferably, hard or soft gelatin) form. Intravenous and
intramuscular formulations are preferably administered in sterile
saline. Of course, one of ordinary skill in the art may modify the
formulations within the teachings of the specification to provide
numerous formulations for a particular route of administration
without rendering the compositions of the present invention
unstable or compromising their therapeutic activity.
[0465] In certain preferred embodiments, the present compositions
are preferably readily water soluble and mixtures of water-soluble
zincs may be used to effect an immediate release/sustained release
pharmaceutical profile. This may maximize immediate effect and
longer duration effect by simply choose the type of salt and
adjusting the ratio of the zinc salt mixture accordingly. Of
course, excipients can be chosen to affect the delivery and
bioequivalence of the zinc salts used. It is well within the
routineer's skill to modify the route of administration and dosage
regimen of a particular compound in order to manage the
pharmacokinetics of the present compounds for maximum beneficial
effect to the patient.
[0466] Formulations containing the compounds of the invention may
take the form of solid, semi-solid, lyophilized powder, or liquid
dosage forms, such as, for example, tablets, capsules, powders,
sustained-release formulations, solutions, suspensions, emulsions,
sup-positories, creams, ointments, lotions, aerosols or the like,
preferably in unit dosage forms suitable for simple administration
of precise dosages.
[0467] The compositions typically include a conventional
pharmaceutical carrier or excipient and may additionally include
other medicinal agents, carriers, and the like. Preferably, the
composition will be about 0.05% to about 75-80% by weight of a zinc
salt compound or compounds according to the invention, with the
remainder consisting of suitable pharmaceutical additives, carriers
and/or excipients. For oral administration, such excipients include
pharmaceutical grades of mannitol, lactose, starch, magnesium
stearate, sodium saccharine, talcum, cellulose, glucose, gelatin,
sucrose, magnesium carbonate, and the like. If desired, the
composition may also contain minor amounts of non-toxic auxiliary
substances such as wetting agents, emulsifying agents, or
buffers.
[0468] Liquid compositions can be prepared by dissolving or
dispersing the compounds (about 0.5% to about 20%), and optional
pharmaceutical additives, in a carrier, such as, for example,
aqueous saline, aqueous dextrose, glycerol, or ethanol, to form a
solution or suspension. For use in oral liquid preparation, the
composition may be prepared as a solution, suspension, emulsion, or
syrup, being supplied either in liquid form or a dried form
suitable for hydration in water or normal saline.
[0469] When the composition is employed in the form of solid
preparations for oral administration, the preparations may be
tablets, granules, powders, capsules or the like. In a tablet
formulation, the composition is typically formulated with
additives, e.g. an excipient such as a saccharide or cellulose
preparation, a binder such as starch paste or methyl cellulose, a
filler, a disintegrator, and other additives typically used in the
manufacture of medical preparations.
[0470] The present invention also contemplates a route of
administration other than an oral route. An injectable composition
for parenteral administration will typically contain the compound
in a suitable i.v. solution, such as sterile physiological salt
solution. The composition may also be formulated as a suspension in
a lipid or phospholipid, in a liposomal suspension, or in an
aqueous emulsion.
[0471] The pharmaceutical compositions of this invention may also
be administered by nasal aerosol or inhalation. Such compositions
are prepared according to techniques well-known in the art of
pharmaceutical formulation and may be prepared as solutions in
saline, employing benzyl alcohol or other suitable preservatives,
absorption promoters to enhance bioavailability, fluorocarbons,
and/or other conventional solubilizing or dispersing agents.
[0472] Methods for preparing such dosage forms are known or will be
apparent to those skilled in the art; for example, see "Remington's
Pharmaceutical Sciences" (17th Ed., Mack Pub. Co., 1985). The
person of ordinary skill will take advantage of favorable
pharmacokinetic parameters of the pro-drug forms of the present
invention, where applicable, in delivering the present compounds to
a patient suffering from a viral infection to maximize the intended
effect of the compound.
[0473] The pharmaceutical compositions according to the invention
may also contain other active ingredients such as proton pump
inhibitors, H.sub.2 blockers, antimicrobial agents, cytoprotective
agents or combination agents. In addition, compounds according to
the present invention may also contain anti-cancer agents (to treat
gastric cancer). Effective amounts or concentrations of each of the
active compounds are to be included within the pharmaceutical
compositions according to the present invention.
[0474] The individual components of such combinations may be
administered either sequentially or simultaneously in separate or
combined pharmaceutical formulations.
[0475] When one or more of the compounds according to the present
invention is used in combination with a second therapeutic agent
active the dose of each compound may be either the same as or
differ from that when the compound is used alone. Appropriate doses
will be readily appreciated by those skilled in the art.
[0476] The following examples are used to describe the present
invention. It is understood that they are merely exemplary and are
understood not to limit the breadth of the invention in any
way.
EXAMPLES
[0477] The stomach produces acid to help break down food, making it
easier to digest. In some cases, stomach acid can actually irritate
the lining of the stomach and the duodenum (top end of the small
intestine). Sometimes the acid "reluxes" upwards and irritates the
lining of the esophagus. Irritation of the lining of the stomach or
the esophagus causes acid indigestion (heartburn) and sometimes
causes ulcers or bleeding.
[0478] We show in this particular application that ZnCl.sub.2 has a
potent inhibitory effect on gastric acid secretion at the cellular
level by abolishing the activity of the gastric
H.sup.+,K.sup.+-ATPase in rat and human gastric glands. We also
demonstrate that addition of micromole concentrations of ZnCl.sub.2
can effectively prevent histamine dependent acid secretion in whole
rat stomachs and through a ZnCl.sub.2 enriched diet.
Material and Methods
[0479] Animals.
[0480] Sprague-Dawley rats 150-250 g (Charles River Laboratory))
were housed in climate- and humidity-controlled, light-cycled
rooms, fed standard chow with free access to water, and handled
according to the humane practices of animal care established by the
Yale Animal Care. Prior to experiments, animals were fasted for
18-24 hours with free access to water.
[0481] Isolation of Rat and Human Gastric Glands.
[0482] Following removal of the stomach, the stomach was opened
longitudinally and the corpus and antrum isolated and sliced into
0.5 cm square sections, and washed with cold Ringer solution to
remove residual food particles. The tissues were transferred to the
stage of a dissecting microscope. Individual glands were isolated
using a hand-dissection technique as described previously.sup.36.
Following isolation, individual isolated glands were allowed to
adhere to cover slips that had been pre-coated with Cell-Tak
(Collaborative Research, Bedford, Mass.) and were transferred to
the stage of an inverted microscope.
[0483] The human tissue was transferred from the OR in a
HEPES-buffered Ringer solution. The tissue was stored on ice and
immediately the isolated glands were dissected as described
above.
[0484] Digital Imaging for Intracellular pH.
[0485] Isolated gastric glands were incubated in a HEPES-buffered
Ringer's solution containing either 10 .mu.mol of the pH-sensitive
dye BCECF-AM
(2',7')-bis-(2-carboxyethyl)-5-(and-6)-carboxy-fluorescin,
aceto-methyl ester (Molecular Probes, Eugene, Oreg.) for 10 minutes
as described previously.sup.37-39. Following dye-loading the
chamber was flushed with a HEPES solution to remove all
non-de-esterfied dye. The perfusion chamber was mounted on the
stage of an inverted microscope (Olympus IX50), which was used in
the epifluorescence mode with a 40.times. objective. BCECF was
successively excited at 440 nm and 490 nm from a monochromator
light source, and the resultant fluorescent signal was monitored at
535 nm using an intensified charge-coupled device camera.
Individual regions of interest were outlined and simultaneously
monitored every 15 sec. during the course of the experiment. A
minimum of 8 cells or regions was selected per gland.
[0486] Proton extrusion by individual parietal cells was monitored
by observing recovery of pH.sub.i after acid loading the cells with
Na.sup.+ free HEPES solution containing 20 mM NH.sub.4Cl. Parietal
cells were subsequently superfused with Na.sup.+ free HEPES, which
abolished all Na.sup.+/H.sup.+ Exchanger (NHE) activity, trapping
H.sup.+ within the cytosol and initiating an immediate drop in
pH.sub.i. Under these conditions, the only potential H.sup.+
extrusion pathway is via the H.sup.+,K.sup.+-ATPase activation.
[0487] The intensity ratio data (490/440) were converted to pH
values using the high K.sup.+/nigericin calibration
technique.sup.40. Intracellular pH recovery rates were calculated
from the same initial starting pH to eliminate the potential
variation in the individual intracellular buffering power of the
cells under the different experimental conditions. All data
including the individual images for all wavelengths were recorded
to the hard disk which allowed us to return to the individual
images after the experiment for further analysis. The recovery
rates are expressed as the .DELTA.pH/min. and were calculated over
the pH range of 6.5-6.8. All chemicals were obtained from Sigma and
Molecular Probes. All data were summarized as means SE and were
analysed by grouping measurements at baseline values.
[0488] Whole Stomach pH Measurements.
[0489] Before the experiments animals were fasted for 24 h to
reduce basal acid secretion to a consistent minimum. Animals were
killed with an overdose of isoflurane and an abdominal incision was
made. The stomach was ligated at the duodenal and esophageal
junction and excised. Then 1 ml of non buffered, isotonic saline
(140 mM) was infused into the lumen of the stomach. This volume did
not distend the stomach, thus avoiding potential stimulation of
acid secretion by stretch. The stomachs were then placed in either
oxygenated HEPES-buffered Ringer solution or in the same solution
containing 100 .mu.M histamine alone or additionally 300 .mu.M
ZnCl.sub.2 (pH 7.4) and maintained at 37.degree. C. After 1 hour
the stomach contents were aspirated and the pH was recorded.
[0490] Oral Zinc Supplementation in Rats.
[0491] These studies were designed to modulate acid secretion by
increasing dietary zinc. In these studies we used an oral
ZnCl.sub.2 solution (zinc chloride in tap water). The animals had
free access to food and the zinc containing water for the duration
of the study, 150 mg/kg/d or 0.5 mg/kg/d ZnCl.sub.2 was added to
the drinking water for 5 days. Animals had free access to water
prior to the experiment and were fed with standard chow until 24
hours before the experiment, at which point they had free access to
ZnCl.sub.2 containing water only. After the 5 days exposure period
and the 24 hour fast, the animals were sacrificed and a total
gastrectomy was performed on the animals. Individual gastric glands
were isolated with the hand dissection technique described
above.
Results
[0492] Histamine Induced Acid Secretion in Human and Rat is
Inhibited by ZnCl.sub.2.
[0493] In the first series pH.sub.i measurements of single parietal
cells within freshly isolated gastric glands were used to measure
H.sup.+,K.sup.+-ATPase activity. The activity of the proton pump
was calculated from the rate of alkalinization of pH.sub.i
(.DELTA.pHV.sub.i/min) after acidification using the NH4Cl prepulse
technique in the absence of sodium and bicarbonate. H.sup.+
extrusion under these conditions depends on the activity of the
H.sup.+,K.sup.+-ATPase, as previously shown.sup.41. In the absence
of any stimulation, only a low rate of pHi recovery was observed
(0.011.+-.0.002 pH.sub.i units/min, n=32 cells from 3 glands from 3
animals; FIG. 1 E). After exposure of the rat gastric glands to
histamine (100 .mu.M) the alkalinization rate increased to
0.051.+-.0.004 pH units/min (n=60 cells from 15 glands from 8
animals; FIG. 1 A). Adding 300 .mu.M ZnCl.sub.2 to the superfusion
bath in the presence of histamine (100 .mu.M) prevented the
stimulatory effect of histamine on the Na.sup.+-independent
pH.sub.i recovery rate (0.0012.+-.0.004 pH units/min) and reduced
it to the same level as seen in the control glands not exposed to
histamine (n=60 cells from 6 glands from 4 animals); FIG. 1 B).
Human gastric glands showed also a robust proton efflux under
histamine stimulation. This effect was abolished by ZnCl.sub.2
(FIG. 1C, D); (n=26 cells, 3 glands). Thus the freshly isolated rat
and human gastric glands showed H.sup.+,K.sup.+-ATPase activity
that could be stimulated by histamine and inhibited by
ZnCl.sub.2.
[0494] ZnCl.sub.2 Inhibits Rat Acid Secretion in a Dose Dependent
Manner.
[0495] ZnCl.sub.2 inhibited H.sup.+ extrusion in a dose dependent
manner (FIG. 2). In this protocol acid secretion was stimulated by
histamine and expressed as .DELTA.pH.sub.i/min. Therefore rat
gastric glands were incubated with 10 .mu.M histamine (15 min) and
histamine was present throughout the entire experiment. To
investigate the inhibitory potency of ZnCl.sub.2 we used different
concentrations (25 .mu.M-300 .mu.M). ZnCl.sub.2 was present during
the entire experiment, including the histamine incubation period of
15 min. 300 .mu.M ZnCl.sub.2 showed a 98% inhibition of proton
extrusion compared to the Histamine induced rate and the
control.
[0496] Fast Onset and Reversible Inhibition of Gastric Acid
Secretion by ZnCl.sub.2.
[0497] There are irreversible (i.g. omeprazole) and reversible
(P-CAB's) acid blockers available.sup.42. We investigate the
reversibility of the inhibitory effect of ZnCl.sub.2 in our in
vitro setting. Thus we stimulated acid secretion with histamine
(100 .mu.M) during the entire experiment. When the intracellular
alkalinization (acid secretion) was observed, ZnCl.sub.2 (300
.mu.M) was added to the superfusion bath. The acid secretion was
abolished after a few seconds (FIG. 3 A). After the removal of
ZnCl.sub.2 in the same experiment acid secretion returns to normal
levels. We were also able to demonstrate reversibility following
incubation and superfusion of parietal cells over 20 min with
ZnCl.sub.2 (300 .mu.M) and histamine (100 .mu.M). After removal of
ZnCl.sub.2 from the superfusion bath the intracellular
alkalinization (proton extrusion) proceeded at normal uninhibited
rate (FIG. 3 B).
[0498] Oral Zinc Supplementation Reduces Basal Rat Gastric Acid
Secretion.
[0499] These studies were designed to modulate acid secretion by
increasing dietary zinc. 150 mg/kg/d or 0.5 mg/kg/d ZnCl.sub.2 were
added to the drinking water for 5 days. The H.sup.+ extrusion rate
was measured with BCECF as described before. Histamine stimulated
parietal cells showed a robust recovery rate (proton extrusion) of
0.051.+-.0.004 (n=120 cells from 15 glands from 8 animals). FIG. 4
shows that the ZnCl.sub.2 (150 mg/kg/d or 0.5 mg/kg/d) in the
drinking water decreased the histamine induced acid secretion
significantly in comparison to the control group with histamine
alone. 150 mg/kg/d: 0.022.+-.0.0045; (n=60 cells from 10 glands
from 3 animals), 0.05 mg/kg/d: 0.034.+-.0.0036; (n=60 cells from 6
glands from 4 animals).
[0500] ZnCl.sub.2 decreased gastric acid production e vivo. To
determine whether ZnCl.sub.2 could inhibit gastric acid secretion
in the whole organ, we examined luminal pH in freshly isolated rat
stomachs after incubation in HEPES or in the same solution the
presence of 100 .mu.M histamine or both, 100 .mu.M histamine and
300 .mu.M ZnCl.sub.2. As illustrated in FIG. 5, in the presence of
histamine mean luminal pH was lower than in control stomach
preparations incubated in HEPES alone (3.15.+-.0.27 vs.
4.59.+-.0.48, n=9 for each, P<0.005). In the presence of
histamine and ZnCl.sub.2 the luminal pH was nearly as high as in
the control group without stimulation although this findings were
not significant (4.54.+-.0.065 vs. 4.59.+-.0.48, n=8 each group,
P>0.005).
[0501] Different Zinc Salts Shows Different Efficacy in Raising
Intraluminal pH.
[0502] Measurements of whole stomach intraluminal pH using a number
of zinc salts according to the present invention were made to
assess effect of salt and concentration on intraluminal pH.
Isolated whole stomach preparations from rats were cannulated at
the esophageal and duodenal junction and perfused in vitro with
37.degree. C. pH 7.4 Ringers solution. The blood perfusate was then
exposed to 100 .mu.M Histamine to induce acid secretion. The lumen
of the stomach was infused with 0.5 cc of non-buffered isotonic
saline. In some studies one of the following zinc salts was added
to the lumen perfusate at a final concentration of 300 .mu.M (zinc
chloride, zinc sulfate, zinc acetate, zinc citrate). The data are
the sum of 5 separate stomachs from 5 separate animals for each of
the columns. Data are the mean of all studies with the standard
error of the mean displayed. Those results appear in attached FIG.
6.
Discussion
[0503] In this study, we examined the dose dependent inhibition of
gastric acid secretion by ZnCl.sub.2 in human and rat gastric
glands. Furthermore we tried to evaluate the onset of effect of
ZnCl.sub.2 and used whole stomach preparation as well as oral Zinc
supplementation to investigate the effect on gastric acid
secretion.
[0504] Acid secretion was induced by the classically known
secretagogue histamine, which led to a robust proton extrusion via
the H.sup.+,K.sup.+-ATPase in comparison to basal acid secretion in
the resting, unstimulated gland (FIG. 1e). In subsequent studies we
examined the inhibitory effects of ZnCl.sub.2 on secretagogue
sensitive gastric acid secretion. We confirmed the inhibitory
potency of ZnCl.sub.2 (300 .mu.M) on histamine induced acid
secretion. ZnCl.sub.2 inhibits acid secretion in the single gastric
gland in a dose dependent manner. ZnCl.sub.2 abolished proton
extrusion to a level comparable to that of the control experiments
in both, human and rat gastric glands (FIG. 1). This dose would be
equivalent to 40 mg supplementation per day in humans. The daily
recommended amount of Zinc intake is 11 mg. In the literature the
amount considered to be toxic is 10 times higher. Therefore 40 mg
of ZnCl.sub.2 as an oral acid blocker would be significantly lower
than reported toxic doses. In addition a similar amount of
ZnCl.sub.2 also prevented acid secretion in ex vivo whole stomach
preparations (FIG. 5). In these experiments ZnCl.sub.2 was applied
to the luminal side of the stomach and it can thus be concluded
that the metal ion is working directly on the
H.sup.+,K.sup.+-ATPase of the parietal cell or enters the cell to
modulate the signalling pathway of acid secretion. It remains
unclear how ZnCl.sub.2 enters the cell. Previous studies described
Zinc entry into the cell through voltage dependent
Ca.sup.2+-channels and/or the HCO.sub.3/Cl.sup.- exchanger on the
basolateral membrane. Orally applied ZnCl.sub.2 confirmed our
previous results. Proton extrusion by ZnCl.sub.2 treated rats was
significant lower than acid secretion by our control group (FIG.
4). The control refers to histamine stimulated. In the figure, zinc
treated glands are still higher than the control (non-histamine
treated) alone.
[0505] As mentioned in the introduction proton pump inhibitors have
a delayed onset of acute action and the full inhibitory effect is
slow requiring several dose cycles. For example omeprazole reaches
only 30% inhibition of acid secretion on the first day of
treatment.sup.43. Our study characterizes the rapid onset of action
ZnCl.sub.2 as well as its reversibility. Faster onset of effect and
increased duration of action would offer improvement for patients
with GERD and other acid related disorders. In fact as shown in
FIG. 5a we were able to inhibit histamine induced acid secretion
during maximal proton extrusion by addition of 300 .mu.M
ZnCl.sub.2On the other hand histamine induced acid secretion
continued after removal of ZnCl.sub.2 from the superfusion bath
demonstrating the reversible nature of ZnCl.sub.2 (FIG. 3 b).
[0506] In summary our findings indicate that ZnCl.sub.2 offer a
more rapid and prolonged inhibition of gastric acid secretion. It
is a reversible and fast acting inhibitor of acid secretion in
single rat and human gastric glands and also in whole stomach
preparations.
[0507] Such treatment may provide significant benefit to patients
with GERD. Future studies investigating the exact mechanism by
which ZnCl.sub.2 inhibits acid secretion are necessary and will
help define its future place in the treatment of acid related
diseases in the clinical setting.
Fundic Region
[0508] In the following examples, it is shown that the fundic
region of the stomach and the fundic glands contain functional acid
secretory proteins. Furthermore, it is shown that the fundic glands
have a sodium and potassium independent protein the proton ATPase
commonly referred to as the Vacuolar H.sup.+-ATPase. The evidence
consists of immunofluorescence data using a antibody directed
against the a Subunit of the H.sup.+-ATPase and functional data
(FIGS. 7-10) in which the extrusion rate of protons from these
cells in the absence of Na and K is measured. Further there is
evidence that this process is amplified in the presence of
histamine a compound that was thought to only influence the gastric
H.sup.+,K.sup.+-ATPase found in the parietal cells in the body of
the stomach. This activity is demonstrated in both the rat model
and in humans in gastric resections taken from patients undergoing
gastric reduction surgery.
Materials and Methods
Animals and Chemicals
[0509] Male Sprague Dawley rats weighing 200-300 g were housed in
climate and humidity controlled, light cycled rooms and fed
standard chow with free access to water. Prior to experiments,
animals were fasted while allowing free access to water for 18-24
hours to reduce basal acid secretion. Following isoflurane
anesthesia the animals were sacrificed, an abdominal incision was
made exposing the stomach. After isolating the esophagus and
duodenal junctures a total gastrectomy was performed with 1-2 cm of
esophagus remaining attached to the gastrectomy. We included the
esophageal juncture to have a common landmark for all fundic
isolations. While holding the esophagus with forceps approximately
3 mm of fundus was removed with 5 mm of the intact esophagus.
Fundic Gland Isolation
[0510] The removed fundic tissue was placed in ice cold
HEPES-buffered Ringer-solution (pH adjusted to 7.4 at 4.degree. C.)
and transferred to the stage of a dissection microscope. The fundic
glands were visualized under the microscope at 50.times.
magnification. Glands adjacent to the esophageal junction were hand
dissected. Following isolation, individual glands were adhered to
cover slips that had been pretreated with the biological adhesive
Cell-Tak (Cell-Tak cell adhesive, BD Biosciences; Bedford,
Mass.)
Immunohistochemistry/Immunofluorescence
[0511] Male Wistar rats (200-250 g) were anesthetized with
pentobarbital i.p. and perfused through the left ventricle with PBS
followed by paraformaldehyde-lysine-periodate (PLP) fixative as
previously described.sup.19a. Stomachs were removed, cleaned from
food residues, and fixed overnight at 4.degree. C. by immersion in
PLP. Stomachs were washed three times with PBS and sections were
cut at a thickness of 5 .mu.m after cryoprotection with 2.3 M in
PBS for at least 12 h. Immunostaining was carried out as described
previously.sup.2a. Sections were incubated with 1% SDS for 5 min.,
washed 3 times with PBS and incubated with PBS containing 1% bovine
serum albumin for 15 min prior to the primary antibody. The primary
antibodies (mouse monoclonal anti human .beta. gastric
H.sup.+,K.sup.+-ATPase (Affinity Bioreagents, CA, USA) diluted 1:50
in PBS and applied overnight at 4.degree. C. Sections were then
washed twice for 5 min with high NaCl PBS (PBS+2.7% NaCl), once
with PBS, and incubated with the secondary antibody (donkey
anti-rabbit Alexa 546, Molecular Probes, Oregon) at a dilution of
1:1000 for 1 h at room temperature. Sections were washed twice with
high NaCl PBS and once with PBS before mounting with VectaMount
(Vector Laboratories, Burlingame, Calif.). The specimens were
viewed with a Nikon E-800 microscope.
ImmunoGold Labeling
[0512] Rats were anaesthetized using 5 ml of a 10% sodium
pentobarbital given i.p. Fixation was done through a left ventricle
cardiac perfusion using PBS and then PLP. The stomach was removed
and fixed in PLP for 4 hours and then transferred to holding buffer
overnight. Frozen and Epon sections of the gastro esophageal
junction were made and slices were taken for gold labeling and
electron microscopy.
Hematoxylin and Eosin Staining
[0513] Rats were anaesthetized using 5 ml of a 10% sodium
pentobarbital given i.p. Fixation was done through a left ventricle
cardiac perfusion using PBS to flush the animal and then Karnovsky
Fixative for 2 hours and then in holding buffer overnight. A
section of the gastro esophageal junction were made and slices
taken for Electron Microscopy morphology of the
H.sup.+,K.sup.+-ATPase protein and Hematoxylin/Eosin staining of
the glands at the gastro esophageal junction.
Measurements of Intracellular pH (pH.sub.i) Measurements of
Isolated Fundic Glands
[0514] Using the same protocol that we developed for isolated
corpus gland perfusion.sup.20a,21a individual fundic glands were
loaded with a 10 .mu.M concentration of the pH sensitive dye
(BCECF, (2'7-bis(2-carbocyethal)-5-(and
6)-carboxyflurorescein-acetomethylester; Molecular Probes, OR USA))
for 15 minutes. Following the loading period, the perfusion chamber
was mounted on the stage of an inverted microscope (Olympus IX50)
attached to a digital imaging system (Universal Imaging Corp;
Dowingtown, Pa.), and perfused with HEPES buffered Ringer-solution
for 5 min at 37.degree. C. to remove any unesterified dye.
Measurements were performed in the epifluorescence mode with
60.times./0.80 and 40.times./0.90 objectives. BCECF was
successively excited at 440.+-.10 nm and 490.+-.10 nm, the
resultant intracellular fluorescent signal was monitored at 535 nm
using an intensified charge-coupled device camera. Data points were
acquired every 15 s. The resulting 490/440 intensity ratio data
were converted to intracellular pH (pH.sub.i) values using the high
K.sup.+/Nigericin calibration technique.sup.22as,22as. Acid
extrusion was monitored in the absence of bicarbonate. The rate of
intracellular alkalinization was measured after using the
NH.sub.4Cl-prepulse technique.sup.22a,23a, which resulted in a
reproducible and sustained intracellular acidification.
Intracellular pH recovery rates (H.sup.+, K.sup.+-ATPase activity)
were measured in Na.sup.+ free HEPES solutions containing: 1) 100
.mu.M histamine 2) 100 .mu.M pentagastrin. 3) 100 .mu.M
acetylcholine 4) 100 .mu.M histamine+omeprazole at 100 .mu.M and
200 .mu.M concentrations.
[0515] Intracellular pH recovery rates were calculated from the
same initial starting pH to eliminate the potential variation in
the individual intracellular buffering power of the cells under the
different experimental conditions. All data including the
individual images for all wavelengths were recorded to the hard
disk which allowed us to return to the individual images after the
experiment for further analysis. The recovery rates are expressed
as the .DELTA.pH.sub.i/min, and were calculated over the pH range
of 6.5-6.9.
[0516] Activation of acid secretion via histamine, acetylcholine or
pentagastrin was induced by preincubation of the glands for 15 min
before the experiment combined with BCECF (100 .mu.M) loading. All
data are summarized as mean.+-.S.E. Significance was determined
using the one-way ANOVA test with p<0.05 considered to be
statistically significant. All chemicals used were obtained from
Sigma and Molecular Probes.
Results
[0517] Immunohistochemical Localization of the H.sup.+,
K.sup.+-ATPase
[0518] Immunohistochemistry using specific antibodies directed
against highly conserved epitopes within either the .alpha. or
.beta. subunits of the gastric H.sup.+, K.sup.+-ATPase identified
specific staining for both subunits in the fundic glands (FIG. 7
A).
Electron Microscopy
[0519] After obtaining Epon sections of fasted rat gastro
esophageal junction Electron Microscopy was done on the gastric
glands that came right after this junction and those we named F1
and used in all of our experiments. FIG. 7 B, C shows the gold tag
localization to the H.sup.+, K.sup.+-ATPase in the parietal cell
from the Fundic gland. We noticed a higher density of staining on
the apical pole of the cell in the secretory or vacuolar
canaliculi. This may correlate with the fundic regions high basal
proton extrusion rates in comparison to the corpus due to the fact
that the protein is always at the membrane in the fundic gland
whereas in the corpus the receptor is inside the secretory
canaliculus until stimulation.
H2 Receptor Staining
[0520] H2 receptor staining was done on both the fundus and corpus
to examine the presence and density of the receptor in both the
areas of the stomach. We found clear basolateral staining in the
corpus glands and could not detect staining in the fundic glands.
These results correlate with the lack of effect by histamine in
stimulating fundic acid secretion. It was clearly seen that the H2
receptor is absent in the glands of the fundus and present in the
corpus (data not shown)
Secretagogue Induced Acid Secretion
[0521] Intracellular pH was measured using the pH sensitive dye
BCECF and monitored continuously using a real time fluorescence
imaging system to identify changes in intracellular pH. Rates of
proton efflux were calculated as .DELTA.pH.sub.i/min using a
technique that was developed in our laboratory for corpus
glands.sup.21a-25a. We measured the change in rate of efflux in the
presence and absence of secretagogues.
Histamine Effect on Fundic and Corpus H.sup.+, K.sup.+-ATPase
[0522] We incubated individual glands with 100 .mu.M histamine for
20 minutes. Histamine was present during the whole superfusion
protocol. In the corpus gland we measured a histamine stimulated
proton extrusion rate of 0.056.+-.0.008 .DELTA.pH.sub.i/min whereas
the basal acid secretion without any secretagogues was
0.011.+-.0.002 .DELTA.pH.sub.i/min (FIG. 8C, D). In comparison to
the corpus the fundus showed even under basal conditions a high
proton extrusion rate (0.039.+-.0.009 .DELTA.pH.sub.i min). This is
similar to the histamine induced acid secretion (0.040.+-.0.0079
.DELTA.pH.sub.i/min, FIG. 8 A, B) This data shows that there is no
effect of histamine on F1 zone glands in comparison to
controls.
Acetylcholine and Fundic Acid Secretion
[0523] In the next series we investigated the functional properties
of fundic glands according to the neuronal stimulation via ACH. In
contrast to histamine there was a noticeable change in proton
extrusion rates after stimulation. Although the controls were still
actively pumping out protons the glands that were stimulated with
100 .mu.M of acetylcholine for 20 minutes during the dye loading
and throughout the perfusion. We determined that acetylcholine
caused an increase in the rate of alkalinization (0.075.+-.0.0015
.DELTA.pH.sub.i/min vs. controls 0.039.+-.0.009
.DELTA.pH.sub.i/min) showing a direct effect of acetylcholine on
fundic acid extrusion (FIG. 9).
Pentagastrin Effect on the F1 Zone
[0524] To determine if gastrin could also activate the fundic
H.sup.+, K.sup.+-ATPase we conducted studies using pentagastrin, a
synthetic peptide containing the entire five terminal amino acids
of gastrin, which is know to cause robust acid secretion in corpus
glands. At a dose of 100 .mu.M pentagastrin we observed
alkalinization rates that were 0.062.+-.0.007 .DELTA.pH.sub.i/min
which was similar to acetylcholine in terms of enhancing the rate
of proton extrusion from fundic cells (FIG. 9).
Inhibitors of Gastric Acid Secretion
[0525] In the next series of studies we tried to determine if the
fundic glands had similar H.sup.+, K.sup.+-ATPase inhibitor
profiles as observed in the corpus. We chose the well characterized
inhibitor of the gastric H.sup.+, K.sup.+-ATPase omeprazole and the
P-CAB (potassium competitive acid blocker) AZD0865..sup.26,27.
Omeprazole Effect on the F1 Zone and Corpus
[0526] As shown in FIG. 10 A, omeprazole did not inhibit acid
secretion using the same concentration that completely inhibited
secretagogue induced acid secretion in the corpus (FIG. 10 B). At
even a higher dose than what normally inhibits acid secretion in
the corpus the fundus continued to extrude protons. The fundic
glands were preincubated with 200 .mu.M omeprazole and 100 .mu.M
histamine and then perfused with omeprazole and histamine
throughout the entire experiment. Alkalinization rates were
0.045.+-.0.002 .DELTA.pH.sub.i/min compared to only histamine
stimulated controls at a rate of 0.042.+-.0.007
.DELTA.pH.sub.i/min. In contrast acid secretion in the corpus
glands was abolished by 200 .mu.mol omeprazole (0.014.+-.0.002
.DELTA.pH.sub.i/min.), (FIG. 10 B).
AZD0865 Effect on the F1 Zone in Comparison to the Corpus:
[0527] Also shown in FIG. 10 (C, D), P-CAB AZD0865 effectively
inhibits acid secretion in the corpus at a 10 .mu.M concentration;
however at that same concentration the F1 zone still has potassium
dependant recovery. In the fundus the intracellular pH increased at
a rate of 0.031.+-.0.006 .DELTA.pH.sub.i/min. In the Corpus at the
same concentration of 10 .mu.M the recovery rate was 0.021.+-.0.008
.DELTA.pH.sub.i/min.
Table 1
[0528] Composition of Solutions Used for Intracellular pH
Measurements in Single Rat Gastric Glands.
[0529] All concentrations are given in Mm. NMDG is
N-Methyl-D-Glucosamine, all solutions were titrated to pH 7.4 at
37.degree. C. using either NaOH or KOH. NMDG was titrated with
HCL.
TABLE-US-00002 TABLE 1 Solution 3: Solution 1: Solution 2: Na.sup.+
- free Solution 4: Standard Na.sup.+ - free HEPES + High K.sup.+
HEPES HEPES NH4Cl calibration NaCl 125 -- -- -- NMDG -- 125 125 125
NH.sub.4Cl -- -- 20 -- KCl 3 3 3 105 MgSO.sub.4 1.2 1.2 1.2 1.2
CaCl.sub.2 1 1 1 1 Glucose 5 5 5 -- HEPES 32.2 32.2 32.2 32.2 pH
7.4 7.4 7.4 7.0
Discussion
[0530] In this study we have provided evidence that the fundic
region of the stomach contains glands that are capable of secreting
acid via the gastric H.sup.+, K.sup.+-ATPase. In our study we have
for the first time characterized the acid secretory properties of
the fundus. We provide morphological, immunohistochemical and
functional evidence for H.sup.+, K.sup.+-ATPase protein activity in
the fundus In our morphological studies we first had to delineate
where the fundus began and ended. As it is understood that this
region begins at the gastro esophageal junction we decided to take
glands from this junction point until the initiation of the greater
curve of the fundus. We took tissue sections from what we called
the F1 zone starting from the gastro esophageal junction and
continued 2 mm distally. We found these glands to be quite
different in shape and also in parietal cell like density. To
confirm that parietal cells in F1 contain H.sup.+, K.sup.+-ATPase
we stained for the .alpha. and .beta. subunit (FIG. 7 shows the
staining for the a subunit).
[0531] During our investigation of secretagogue induced fundic acid
secretion we were able to demonstrate that histamine is not the
most potent stimulator of acid secretion in the fundus as it is in
the corpus. In fact, very little difference was seen in glands that
were not stimulated compared to those stimulated with histamine
(FIGS. 8 and 9). This result was confirmed by a lack of staining
for the H2 receptor in the fundus (data not shown). Acetylcholine
was the most robust of the three secretagogues in the fundus, which
may relate to the close proximity of the vagal nerve to the fundic
region. As this section of the stomach stretches when food is
present, there is vagal stimulation.sup.28a-30a with associated
acetylcholine secretion. This finding is especially important when
considering clinical problems in obese patients who have gastro
esophageal reflux disease (GERD). This can be correlated to the
benefits patients with sever ulcer disease gain when undergoing a
vagotomy after not gaining relief from medical
management.sup.31a-33a.
[0532] Another interesting finding is the lack of inhibition by
omeprazole on fundic acid secretion although they are
immunoreactive with antibodies for the gastric H.sup.+,
K.sup.+-ATPase. We were unable to inhibit basal or secretagogue
(histamine) induced fundic acid secretion with the proton pump
inhibitor omeprazole (FIG. 10 A) at doses that were double that
which effectively eliminated all acid secretion in the corpus (FIG.
10 B). These findings are in direct contrast to our data (FIG. 8 C.
D).sup.20a, 21a, 24a, 25a and others findings in glands from the
corpus.sup.34a-38a This finding has an interesting clinical
correlate in that there is an increasing number of patients
suffering from GERD that are not effectively treated with
PPI's..sup.39a One possible explanation for the lack of omeprazole
sensitivity could be that as omeprazole needs to be acid activated,
a lack of a canaliculi like space would prevent the concentration
of acid and potentially prevent the acid activation of the drug. As
shown in FIG. 7 B, C using immunogold tag labeling we see that
there is indeed a secretory space and that the pumps appear to line
the apical surface of this space. With previous theories of acid
secretion came the many dictums for surgical and medical modalities
of treatment which focused on the corpus. Of interest is the recent
incidence of GERD like symptoms in gastric bypass patients who
following the procedure are left only with a small part of the
fundus, and little to no functional corpus. In those symptomatic
patients there has been little to no success using classical
PPI's.sup.40a which now can be possibly be explained by our recent
findings. Our findings demonstrate that the fundic region of the
stomach is much more than a holding area and in fact can secrete
acid in response to secretagogue stimulation, furthermore the
H.sup.+, K.sup.+-ATPase found in this segment appears insensitive
to omeprazole. These results can lead to important new targets for
patients that are PPI resistant or have recurrent reflux symptoms
in the presence of PPI therapy.
Results:
[0533] Fundic glands showed a distinct morphology compared to
corpus glands (elongated and lacking typical bulging parietal
cells). Immunofluorescence (.alpha. and .beta. subunit of the
H.sup.+, K.sup.+-ATPase) and immunogold labeling (B subunit) were
both positive in the fundic region. Fundic gland proton extrusion
rates were stimulated by gastrin and acetylcholine but were not
influenced by histamine. Finally acid secretion of stimulated
fundic glands could not be inhibited by the H.sup.+, K.sup.+-ATPase
inhibitor omeprazole.
Conclusion
[0534] The fundic region of the stomach secreted acid via the
H.sup.+,K.sup.+ ATPase, and was not sensitive to proton pump
inhibitors. Our findings demonstrate that the fundic region of the
stomach is much more than a holding area and in fact can secrete
acid in response to secretagogue stimulation, except histamine.
[0535] The digestion of food by the stomach requires a complex
combination of hormonal and neuronal events. Generally it has been
thought that the corpus or body of the stomach secretes acid via
the parietal cells and the antrum secretes bicarbonate to
neutralize the digestate by raising the pH of the stomach
contents.sup.1a-9a. During this process the peristaltic movements
of the stomach result in contractions that push the food upward
into the fundic section of the stomach where it transits before
exiting into the small intestine.sup.10a. In this model of
digestion the fundus acts only as a holding zone and is not
involved in acid secretion.sup.4a,11a. Classical gastric acid
secretion in the corpus occurs when the H.sup.+, K.sup.+-ATPase
gets stimulated by secretagogues and begins to secrete protons into
the secretory canaliculus after being trafficked to the apical
membrane from their cytoplasmic tubulovesicles.sup.12a. The
parietal cell has at least three activating receptors on its
basolateral membrane, i.e. histamine H2, acetylcholine M3 and
Gastrin CCK-B. It is well accepted that the H2 receptor couples to
Gs to activated adenylate cyclase producing cAMP and subsequent
activation of cAMP dependant protein kinase. The acetylcholine and
gastrin receptor couple through a non Gs system probably Gq to
activate phospholipase C producing IP3 and diacylglycerol.
Acetylcholine releases intracellular Ca.sup.2+ and gastrin
activating protein kinase C.sup.13a. After this cascade of
intracellular events the parietal cell extrudes protons via the
H.sup.+, K.sup.+-ATPase pump which exchanges intracellular H.sup.+
ions for extracellular K.sup.+ ions in an electroneutral
ratio.sup.14a.
[0536] Recent observations in patients who have undergone gastric
bypass surgery present an interesting paradigm, namely that only
left with a small fundic region postoperatively, they still have
acid secretion which in some patients leads to reflux symptoms,
ulcers and enteric content leaks.sup.15a. Of note is that many of
these patients have little success in abating the symptoms while on
proton pump inhibitor (PPI) therapy.sup.16a-18a. From these initial
clinical observations we raised the question: does the fundus play
a role in the production of acid, and how similar are its
properties to corpus secretory proteins. We also were interested in
determining fundic sensitivity to classical secretagogues and
therefore conducted studies using histamine, pentagastrin, and
acetylcholine.
[0537] In the present experiment we have investigated the acid
secretory properties of the rat fundus under resting and
secretagogue stimulated states, furthermore we elucidate fundic
response to an inhibitory drug of acid secretion. Our data
demonstrate that the fundic region is an active secretory zone in
the stomach and contains a gastric H.sup.+, K.sup.+-ATPase that can
be stimulated by secretagogues but appears to be insensitive to
omeprazole. Zinc therapy according to the present invention is a
means to regulate acid release in the fundus region.
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