U.S. patent application number 11/102630 was filed with the patent office on 2005-08-11 for monoamine oxidase (mao) inhibitors and uses thereof.
This patent application is currently assigned to Regent Court Technologies. Invention is credited to Burton, Harold R., DeLorenzo, Robert J., Williams, Jonnie R..
Application Number | 20050176777 11/102630 |
Document ID | / |
Family ID | 22209450 |
Filed Date | 2005-08-11 |
United States Patent
Application |
20050176777 |
Kind Code |
A1 |
Williams, Jonnie R. ; et
al. |
August 11, 2005 |
Monoamine oxidase (MAO) inhibitors and uses thereof
Abstract
The present invention provides a group of tobacco alkaloids,
tobacco extract, Yerbamat extract, and an extract of chewing gum
and lozenges which are modulators of monoamine oxidase (MAO)
activity (i.e., compounds and substances which inhibit MAO enzyme
and prevent its biological activity). The MAO inhibitors of the
present invention can cause an increase in the level of
norepinephrine, dopamine, and serotonin in the brain and other
tissues, and thus can cause a wide variety of pharmacological
effects mediated by their effects on these compounds. The MAO
inhibitors of the present invention are useful for a variety of
therapeutic applications, such as the treatment of depression,
disorders of attention and focus, mood and emotional disorders,
Parkinson's disease, extrapyramidal disorders, hypertension,
substance abuse, smoking substitution, anti-depression therapy,
eating disorders, withdrawal syndromes, and the cessation of
smoking.
Inventors: |
Williams, Jonnie R.;
(Manakin-Sabot, VA) ; DeLorenzo, Robert J.;
(Richmond, VA) ; Burton, Harold R.; (Lexington,
KY) |
Correspondence
Address: |
BANNER & WITCOFF
1001 G STREET N W
SUITE 1100
WASHINGTON
DC
20001
US
|
Assignee: |
Regent Court Technologies
Town and Country
MO
|
Family ID: |
22209450 |
Appl. No.: |
11/102630 |
Filed: |
April 11, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11102630 |
Apr 11, 2005 |
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10396319 |
Mar 26, 2003 |
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10396319 |
Mar 26, 2003 |
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10042164 |
Jan 11, 2002 |
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6569470 |
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10042164 |
Jan 11, 2002 |
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09325852 |
Jun 4, 1999 |
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6350479 |
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60088117 |
Jun 5, 1998 |
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Current U.S.
Class: |
514/343 |
Current CPC
Class: |
A61P 25/00 20180101;
A23G 4/068 20130101; A61P 25/34 20180101; A23L 33/105 20160801;
A61P 25/30 20180101; A61P 25/04 20180101; A61K 36/185 20130101;
A61P 25/24 20180101; A23F 3/34 20130101; A61P 25/14 20180101; A61K
31/4439 20130101; A61P 43/00 20180101; A61P 25/22 20180101; A61K
36/81 20130101; A61K 36/185 20130101; A61K 36/81 20130101; A61P
25/28 20180101; A61K 2300/00 20130101; A61P 25/16 20180101; A61K
2300/00 20130101 |
Class at
Publication: |
514/343 |
International
Class: |
A61K 031/4439 |
Claims
What is claimed is:
1. A method of treating drug withdrawal syndromes or drug
dependence disorders comprising administering to a mammal in need
thereof an effective amount of an active agent in a
pharmaceutically acceptable carrier, diluent or vehicle, wherein
said active agent is selected from the group consisting of
anabasine, anatabine, and nornicotine.
2. The method of claim 1, wherein the mammal is human.
3. The method of claim 2, wherein the active agent is administered
in an amount effective to inhibit monoamine oxidase (MAO)
activity.
4. The method of claim 3, wherein the inhibition of MAO activity is
asymptotic.
5. The method of claim 1, wherein the active agent is
anabasine.
6. The method of claim 1, wherein the active agent is
anatabine.
7. The method of claim 1, wherein the active agent is
nornicotine.
8. The method of claim 4, wherein the active agent is administered
in an amount effective to inhibit monoamine oxidase A (MAO A)
activity.
9. The method of claim 4, wherein the active agent is administered
in an amount effective to inhibit monoamine oxidase B (MAO B)
activity.
10. The method of claim 1, wherein the drug withdrawal syndrome or
drug dependence disorder involves a dependence on at least one of
alcohol, opioids, amphetamines, cocaine, tobacco, and cannabis.
11. A method of treating tobacco withdrawal syndrome or tobacco
dependence disorder comprising administering to a mammal in need
thereof an effective monoamine oxidase (MAO) inhibiting amount of
anabasine in a pharmaceutically acceptable carrier, diluent or
vehicle.
12. The method of claim 11, wherein the mammal is human.
13. The method of claim 12, wherein the inhibition of MAO is
asymptotic.
14. The method of claim 12, wherein the anabasine is administered
in an amount effective to inhibit monoamine oxidase A (MAO A)
activity.
15. The method of claim 12, wherein the anabasine is administered
in an amount effective to inhibit monoamine oxidase B (MAO B)
activity.
16. A method of treating tobacco withdrawal syndrome or tobacco
dependence disorder comprising administering to a mammal in need
thereof an effective monoamine oxidase (MAO) inhibiting amount of
anatabine in a pharmaceutically acceptable carrier, diluent or
vehicle.
17. The method of claim 16, wherein the mammal is human.
18. The method of claim 17, wherein the inhibition of MAO is
asymptotic.
19. The method of claim 17, wherein the anatabine is administered
in an amount effective to inhibit monoamine oxidase A (MAO A)
activity.
20. The method of claim 17, wherein the anatabine is administered
in an amount effective to inhibit monoamine oxidase B (MAO B)
activity.
21. A method of treating tobacco withdrawal syndrome or tobacco
dependence disorder comprising administering to a mammal in need
thereof an effective monoamine oxidase (MAO) inhibiting amount of
nomicotine in a pharmaceutically acceptable carrier, diluent or
vehicle.
22. The method of claim 21, wherein the mammal is human.
23. The method of claim 22, wherein the inhibition of MAO is
asymptotic.
24. The method of claim 22, wherein the nornicotine is administered
in an amount effective to inhibit monoamine oxidase A (MAO A)
activity.
25. The method of claim 22, wherein the nornicotine is administered
in an amount effective to inhibit monoamine oxidase B (MAO B)
activity.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a division of application Ser. No.
10/396,319, filed Mar. 26, 2003, which is a division of application
Ser. No. 10/042,164, filed Jan. 11, 2002, now U.S. Pat. No.
6,569,470, which is a division of application Ser. No. 09/325,852,
filed Jun. 4, 1999, now U.S. Pat. No. 6,350,479, which claims
priority under 35 U.S.C. .sctn. 119(e) to Provisional Application
No. 60/088,117, filed Jun. 5, 1998.
FIELD OF THE INVENTION
[0002] The present invention relates to the novel use of compounds
and substances which are capable of modulating monoamine oxidase
(MAO) activity by inhibiting the MAO enzyme. The present invention
also relates to MAO inhibitors and their therapeutic use as a drug
or dietary supplement in the treatment of various conditions or
disorders, including psychiatric and neurological illnesses. More
particularly, the present invention relates to the therapeutic use
of tobacco alkaloids, Yerbamat (Ilex paraguariensis) extract, or
tobacco extracts to inhibit MAO activity to provide a treatment for
various disorders or conditions.
BACKGROUND OF THE INVENTION
[0003] By inhibiting MAO activity, MAO inhibitors can regulate the
level of monoamines and their neurotransmitter release in different
brain regions and in the body (including dopamine, norepinephrine,
and serotonin). Thus, MAO inhibitors can affect the modulation of
neuroendocrine function, respiration, mood, motor control and
function, focus and attention, concentration, memory and cognition,
and the mechanisms of substance abuse. Inhibitors of MAO have been
demonstrated to have effects on attention, cognition, appetite,
substance abuse, memory, cardiovascular function, extrapyramidal
function, pain and gastrointestinal motility and function. The
distribution of MAO in the brain is widespread and includes the
basal ganglia, cerebral cortex, limbic system, and mid and
hind-brain nuclei. In the peripheral tissue, the distribution
includes muscle, the gastrointestinal tract, the cardiovascular
system, autonomic ganglia, the liver, and the endocrinic system.
The present invention overcomes the problems and limitations of the
prior art by providing methods and systems.
[0004] MAO inhibition by other inhibitors have been shown to
increase monoamine content in the brain and body. Regulation of
monoamine levels in the body have been shown to be effective in
numerous disease states including depression, anxiety, stress
disorders, diseases associated with memory function, neuroendocrine
problems, cardiac dysfunction, gastrointestinal disturbances,
eating disorders, hypertension, Parkinson's disease, memory
disturbances, and withdrawal symptoms.
[0005] It has been suggested that cigarette smoke may have
irreversible inhibitory effect towards monoamine oxidase (MAO). A.
A. Boulton, P. H. Yu and K. F. Tipton, "Biogenic Amine Adducts,
Monoamine Oxidase Inhibitors, and Smoking," Lancet, 1(8577):
114-155 (Jan. 16, 1988), reported that the MAO-inhibiting
properties of cigarette smoke may help to explain the protective
action of smoking against Parkinson's disease and also observed
that patients with mental disorders who smoke heavily do not
experience unusual rates of smoking-induced disorders. It was
suggested that smoking, as an MAO inhibitor, may protect against
dopaminergic neurotoxicity that leads to Parkinson's disease and
that the MAO-inhibiting properties of smoking may result in an
anti-depressive effect in mental patients.
[0006] L. A. Carr and J. K. Basham, "Effects of Tobacco Smoke
Constituents on MPTP Induced Toxicity and Monoamine Oxidase
Activity in the Mouse Brain," Life Sciences, 48:1173-1177 (Jan. 16,
1991), found that nicotine, 4-phenylpyridine and hydrazine
prevented the decrease in dopamine metabolite levels induced by
1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in mice, but
there was no significant effect on dopamine levels. Because tobacco
smoke particulate matter caused a marked inhibition of MAO A and
MAO B activity when added in vitro, it was suggested that one or
more unidentified substances in tobacco smoke are capable of
inhibiting brain MAO and perhaps altering the formation of the
active metabolite of MPTP.
[0007] J. S. Fowler, N. D. Volkow, G. J. Wang, N. Pappas, and J.
Logan, "Inhibition of Monoamine Oxidase B in the Brain of Smokers,"
Nature (Lond), 379(6567):733-736 (Feb. 22, 1996), found that the
brains of living smokers showed a 40% decrease in the level of MAO
B relative to non-smokers or former smokers. MAO inhibition was
also reported as being associated with decreased production of
hydrogen peroxide.
[0008] It has also been suggested that nicotine may not be the only
constituent of tobacco responsible for tobacco addiction. J.
Stephenson, "Clues Found to Tobacco Addiction," Journal of the
American Medical Association, 275(16): 1217-1218 (Apr. 24, 1996),
discussing the work of Fowler, et al., pointed out that the brains
of living smokers had less MAO B compared with the brains of
nonsmokers or former smokers. MAO B is an enzyme involved in the
breakdown of dopamine, which is a pleasure-enhancing
neurotransmitter. The results suggested that the inhibition of MAO
B in the brains of smokers may make nicotine more addictive by
slowing down the breakdown of dopamine, thereby boosting its
levels. The findings provided an explanation as to why cigarette
smokers were less susceptible to developing Parkinson's disease.
Further, the findings suggested that MAO inhibitors could be used
for smoking cessation.
[0009] K. R. R. Krishnan, "Monoamine Oxidase Inhibitors," The
American Psychiatric Press Textbook of Pharmacology, American
Psychiatric Press, Inc., Washington, D.C. 1995, pp. 183-193,
suggest various uses for monoamine oxidase inhibitors. The uses
include atypical depression, major depression, dysthymia,
melancholia, panic disorder, bulimia, atypical facial pain, anergic
depression, treatment-resistant depression, Parkinson's disease,
obsessive-compulsive disorder, narcolepsy, headache, chronic pain
syndrome, and generalized anxiety disorder.
[0010] D. Nutt and S. A. Montgomery, "Moclobemide in the Treatment
of Social Phobia," Int. Clin. Psychopharmacol, 11 Suppl. 3: 77-82
(Jun. 11, 1996), reported that moclobemide, a reversible MAO
inhibitor, may be effective in the treatment of social phobia.
[0011] I. Berlin, et al., "A Reversible Monoamine Oxidase A
Inhibitor (Moclobemide) Facilitates Smoking Cessation and
Abstinence in Heavy, Dependent Smokers," Clin. Pharmacol. Ther.,
58(4): 444-452 (October 1995), suggested that a reversible MAO A
inhibitor can be used to facilitate smoking cessation.
[0012] U.S. Pat. No. 3,870,794 discloses the administering of small
quantities of nicotine and nicotine derivatives to mammals,
including humans, to reduce anger and aggressiveness and to improve
task performance.
[0013] U.S. Pat. No. 5,276,043 discloses the administering of an
effective amount of certain anabasine compounds, certain
unsaturated anabasine compounds, or unsaturated nicotine compounds
to treat neurodegenerative diseases.
[0014] U.S. Pat. No. 5,516,785 disclose a method of using
anabasine, and DMAB anabasine for stimulating brain cholinergic
transmission and a method for making anabasine.
[0015] U.S. Pat. Nos. 5,594,011, 5,703,100, 5,705,512, and
5,723,477 disclose modulators of acetylcholine receptors.
[0016] Known irreversible MAO inhibitors also inhibit MAO in the
stomach and liver as well as the brain. As a result, their use has
been limited because hypertensive crisis may occur when certain
types of food (for example, fermented foods) are ingested, thereby
creating an adverse drug-food interaction. Tyramine, which has a
pressor action and which is normally broken down by the MAO
enzymes, can be present in certain foods.
[0017] Thus, it would be desirable to provide MAO inhibitors which
are effective, but less potent (i.e., those which provide an
asymptotic effect on MAO inhibition) than known MAO inhibitors, for
the treatment-of various conditions and disorders. It would also be
desirable to provide MAO inhibitors which are easily synthesized
and which could be provided to patients as an "over the counter"
medication or dietary supplement.
BRIEF SUMMARY OF THE INVENTION
[0018] The present invention relates to the discovery that certain
tobacco alkaloids or extracts, a certain tea plant extract, and a
certain extract of tobacco extract-containing chewing gum and
lozenges provide MAO-inhibiting effects. The present invention also
relates to the use of these compounds or substances in the
treatment of certain conditions and disorders in mammals, including
humans.
[0019] The compounds and substances of the present invention are
capable of inhibiting MAO activity in mammalian brain and
peripheral tissue. These compounds and substances act by increasing
the concentration of monoamine compounds (norepinephrine, dopamine,
and serotonin) in the body and brain.
[0020] The present invention provides a method of treating certain
medical, psychiatric and/or neurological conditions or disorders.
In a first embodiment of the invention, the method comprises
administering a MAO-inhibiting effective amount of anabasine,
anatabine or nornicotine to a mammal, particularly a human, for the
treatment of medical, psychiatric and/or neurological conditions
and disorders such as, but not limited to, Alzheimer's disease,
Parkinson's disease, major depression, minor depression, atypical
depression, dysthymia, attention deficit disorder, hyperactivity,
conduct disorder, narcolepsy, social phobia, obsessive-compulsive
disorder, atypical facial pain, eating disorders, drug withdrawal
syndromes and drug dependence disorders, including dependence from
alcohol, opioids, amphetamines, cocaine, tobacco, and cannabis
(marijuana), melancholia, panic disorder, bulimia, anergic
depression, treatment-resistant depression, headache, chronic pain
syndrome, generalized anxiety disorder, and other conditions in
which alteration of MAO activity could be of therapeutic value.
[0021] In a second embodiment of the invention, the method
comprises administering a MAO-inhibiting effective amount of an
extract of Yerbamat (Ilex paraguariensis) tea plant to a mammal,
particularly a human, for the treatment of medical, psychiatric
and/or neurological conditions and disorders such as, but not
limited to, Alzheimer's disease, Parkinson's disease, major
depression, minor depression, atypical depression, dysthymia,
attention deficit disorder, hyperactivity, conduct disorder,
narcolepsy, social phobia, obsessive-compulsive disorder, atypical
facial pain, eating disorders, drug withdrawal syndromes and drug
dependence disorders, including dependence from alcohol, opioids,
amphetamines, cocaine, tobacco, and cannabis (marijuana),
melancholia, panic disorder, bulimia, anergic depression,
treatment-resistant depression, headache, chronic pain syndrome,
generalized anxiety disorder, and other conditions in which
alteration of MAO activity could be of therapeutic value.
[0022] In a third embodiment of the invention, the method comprises
administering a MAO-inhibiting effective amount of a tobacco
extract to a mammal, particularly a human, for the treatment of
medical, psychiatric and/or neurological conditions and disorders
such as, but not limited to, Alzheimer's disease, Parkinson's
disease, major depression, minor depression, atypical depression,
dysthymia, attention deficit disorder, hyperactivity, conduct
disorder, narcolepsy, social phobia, obsessive-compulsive disorder,
atypical facial pain, eating disorders, drug withdrawal syndromes
and drug dependence disorders, including dependence from alcohol,
opioids, amphetamines, cocaine, tobacco, and cannabis (marijuana),
melancholia, panic disorder, bulimia, anergic depression,
treatment-resistant depression, headache, chronic pain syndrome,
generalized anxiety disorder, and other conditions in which
alteration of MAO activity could be of therapeutic value.
[0023] In a fourth embodiment of the invention, the method
comprises administering a MAO-inhibiting effective amount of an
extract of gum and lozenges formulated with tobacco extract to a
mammal, particularly a human, for the treatment of medical,
psychiatric and/or neurological conditions and disorders such as,
but not limited to, Alzheimer's disease, Parkinson's disease, major
depression, minor depression, atypical depression, dysthymia,
attention deficit disorder, hyperactivity, conduct disorder,
narcolepsy, social phobia, obsessive-compulsive disorder, atypical
facial pain, eating disorders, drug withdrawal syndromes and drug
dependence disorders, including dependence from alcohol, opioids,
amphetamines, cocaine, tobacco, and cannabis (marijuana),
melancholia, panic disorder, bulimia, anergic depression,
treatment-resistant depression, headache, chronic pain syndrome,
generalized anxiety disorder, and other conditions in which
alteration of MAO activity could be of therapeutic value.
BRIEF DESCRIPTION OF THE DRAWINGS
[0024] FIG. 1 shows a plot of MAO inhibition versus time for
anabasine.
[0025] FIG. 2 shows the inhibition of MAO A and MAO B for
anabasine.
[0026] FIG. 3 shows a plot of MAO inhibition versus time for
anatabine.
[0027] FIG. 4 shows the inhibition of MAO A and MAO B for
anatabine.
[0028] FIG. 5 shows a plot of MAO inhibition versus time for
nornicotine.
[0029] FIG. 6 shows the inhibition of MAO A and MAO B for
nornicotine.
[0030] FIG. 7 shows a plot of MAO inhibition versus time for
Yerbamat.
[0031] FIG. 8 shows the inhibition of MAO A and MAO B for
Yerbamat.
[0032] FIG. 9 shows a plot of MAO inhibition versus time for
tobacco extract.
[0033] FIG. 10 shows the inhibition of MAO A and MAO B for tobacco
extract.
[0034] FIG. 11 shows a plot of MAO inhibition versus time for
GUMSMOKE.
[0035] FIG. 12 shows a plot of MAO inhibition versus time for a
lozenge extract.
DETAILED DESCRIPTION OF THE INVENTION
[0036] MAO is an important enzyme that plays a major role in the
metabolic transformation of catecholamines and serotonin.
Neurotransmitters from this group are metabolized by MAO, and thus
their effect is decreased at their receptor cites. MAO is important
for the regulation of the levels of dopamine, norepinephrine and
serotonin.
[0037] Accordingly, inhibition of this major enzyme system will
have major effects on the functions regulated by this compound.
[0038] In a first embodiment of the invention, the method comprises
administering a MAO-inhibiting effective amount of anabasine,
anatabine or nomicotine to a mammal, particularly a human, for the
treatment of medical, psychiatric and/or neurological conditions
and disorders such as, but not limited to, Alzheimer's disease,
Parkinson's disease, major depression, minor depression, atypical
depression, dysthymia, attention deficit disorder, hyperactivity,
conduct disorder, narcolepsy, social phobia, obsessive compulsive
disorder, atypical facial pain, eating disorders, drug withdrawal
syndromes and drug dependence disorders, including dependence from
alcohol, opioids, amphetamines, cocaine, tobacco, and cannabis
(marijuana), melancholia, panic disorder, bulimia, anergic
depression, treatment-resistant depression, headache, chronic pain
syndrome, generalized anxiety disorder, and other conditions in
which alteration of MAO activity could be of therapeutic value.
[0039] Anabasine, anatabine and nornicotine are minor tobacco
alkaloids. These compounds are commercially available. However,
they may be synthesized according to known techniques or extracted
directly from tobacco itself.
[0040] Preferably, anatabine is synthesized according to the method
disclosed by N. M. Deo and P. A. Crooks, "Regioselective Alkylation
of N-(diphenylmethylidine)-3-(aminomethylpyridine: A Simple Route
to Minor Tobacco Alkaloids and Related Compounds," 1137-1141 (11
Dec. 1995), which is incorporated herein by reference.
[0041] In addition, nornicotine is preferably synthesized according
to the method disclosed by S. Brandange and L. Lindblom, "N-Vinyl
as N--H Protecting Group: A Convenient Synthesis of Myosmine," Acta
Chem. Scand., B30, No. 1, p. 93 (1976), which is also incorporated
herein by reference.
[0042] In a second embodiment of the invention, the method
comprises administering a MAO-inhibiting effective amount of an
extract of Yerbamat (Ilex paraguariensis) tea plant to a mammal,
particularly a human, for the treatment of medical, psychiatric
and/or neurological conditions and disorders such as, but not
limited to, Alzheimer's disease, Parkinson's disease, major
depression, minor depression, atypical depression, dysthymia,
attention deficit disorder, hyperactivity, conduct disorder,
narcolepsy, social phobia, obsessive-compulsive disorder, atypical
facial pain, eating disorders, drug withdrawal syndromes and drug
dependence disorders, including dependence from alcohol, opioids,
amphetamines, cocaine, tobacco, and cannabis (marijuana),
melancholia, panic disorder, bulimia, anergic depression,
treatment-resistant depression, headache, chronic pain syndrome,
generalized anxiety disorder, and other conditions in which
alteration of MAO activity could be of therapeutic value.
[0043] The Yerbamat extract may be prepared by shredding the
Yerbamat materials, mixing the shredded materials with a
water/ethanol (for example, 1/1 by volume) solution in a mixture of
about four leaves per 10 ml of the water/ethanol mixture,
extracting with continuous stirring, and then removing the solution
from the Yerbamat residue. The residue can then be further
extracted two more times with the same volume of water/ethanol
mixture, and then the extracts may be combined and filtered to
remove the particulate Yerbamat materials. The combined extracts
may then be subject to vacuum evaporation to yield the Yerbamat
extract.
[0044] In a third embodiment of the invention, the method comprises
administering a MAO-inhibiting effective amount of a tobacco
extract to a mammal, particularly a human, for the treatment of
medical, psychiatric and/or neurological conditions and disorders
such as, but not limited to, Alzheimer's disease, Parkinson's
disease, major depression, minor depression, atypical depression,
dysthymia, attention deficit disorder, hyperactivity, conduct
disorder, narcolepsy, social phobia, obsessive-compulsive disorder,
atypical facial pain, eating disorders, drug withdrawal syndromes
and drug dependence disorders, including dependence from alcohol,
opioids, amphetamines, cocaine, tobacco, and cannabis (marijuana),
melancholia, panic disorder, bulimia, anergic depression,
treatment-resistant depression, headache, chronic pain syndrome,
generalized anxiety disorder, and other conditions in which
alteration of MAO activity could be of therapeutic value.
[0045] The tobacco extract may be prepared by shredding tobacco
leaves (for example, processed tobacco obtained from STAR TOBACCO,
INC.), mixing the shredded leaves with a water/ethanol (for
example, 1/1 by volume) solution in a mixture of about four leaves
per 10 ml of the water/ethanol mixture, extracting with continuous
stirring, and then removing the solution from the tobacco residue.
The residue can then be further extracted two more times with the
same volume of water/ethanol mixture, and then the extracts may be
combined and filtered to remove the particulate tobacco leaf
material. The combined extracts may then be subject to vacuum
evaporation to yield the tobacco extract.
[0046] In a fourth embodiment of the invention, the method
comprises administering a MAO-inhibiting effective amount of an
extract of chewing gum and lozenges formulated with tobacco extract
to a mammal, particularly a human, for the treatment of medical,
psychiatric and/or neurological conditions and disorders such as,
but not limited to, Alzheimer's disease, Parkinson's disease, major
depression, minor depression, atypical depression, dysthymia,
attention deficit disorder, hyperactivity, conduct disorder,
narcolepsy, social phobia, obsessive-compulsive disorder, atypical
facial pain, eating disorders, drug withdrawal syndromes and drug
dependence disorders, including dependence from alcohol, opioids,
amphetamines, cocaine, tobacco, and cannabis (marijuana),
melancholia, panic disorder, bulimia, anergic depression,
treatment-resistant depression, headache, chronic pain syndrome,
generalized anxiety disorder, and other conditions in which
alteration of MAO activity could be of therapeutic value.
[0047] The chewing gum and lozenges extract may be prepared by
extracting five slices of GUMSMOKE chewing gum and NICOMINT
lozenges (obtained from STAR TOBACCO, INC.), which are formulated
with tobacco extract, with distilled water (50 ml) at room
temperature for 12 hours, and then removing the undissolved gum
substance by filtration.
[0048] The above compounds and substances were evaluated for their
MAO inhibiting activity. Test results surprisingly showed that the
compounds and substances of the present invention all provided MAO
inhibition. It was also discovered that the MAO inhibiting effects
had a different character than for known MAO inhibitors in that
they reached an asymptotic or ceiling effect, so that further
increases in the dose beyond maximal inhibition did not produce any
further increase in the MAO inhibition. This asymptotic effect
would provide many benefits. For example, the problems associated
with previously known, irreversible MAO inhibitors, such as
hypertensive effects, can be avoided. Furthermore, the inventive
MAO inhibitors may be provided as an "over the counter" drug or
dietary supplement in view of its safety and efficacy.
[0049] The MAO inhibitors of the present invention may be provided
in forms well known to one skilled in the art. They may be
formulated in a pharmaceutically acceptable carrier, diluent or
vehicle and administered in effective amounts. They may be provided
in the form of a capsule, pill, tablets, lozenge, gum, troches,
suppositories, powder packets or the like.
[0050] The determination of the effective amounts for a given
treatment can be accomplished by routine experimentation and is
also well within the ordinary skill in the art.
EXAMPLES
[0051] To determine the effectiveness of compounds and substances
of the present invention, experiments were conducted as
follows:
[0052] MAO Reaction:
[0053] The MAO activities of the compounds and substances were
determined using standard reaction conditions as described in Halt,
A., et al., Analytical Biochemistry, 244:384-392 (1997).
[0054] Tissue Preparation:
[0055] Liver samples from cow or rat were obtained immediately
after sacrifice. Liver was homogenized in a Polytron mechanical
homogenizer in a ratio of 1 gram of liver to 1 ml of potassium
phosphate buffer (0.2 M at pH of 7.6). Large membranes were removed
by low speed centrifugation at 1000.times.g for 15 minutes. The
supernatant was removed from the pellet and used immediately for
MAO activity assays or stored at 0 degrees Centigrade. Protein
levels were determined in the liver homogenate by the Bradford
protein reaction.
[0056] Reaction Conditions:
[0057] The standard reaction conditions were developed as a
modification of the spectrophotometric assay using standard
conditions (Halt, A., et al., Analytical Biochemistry, 244:384-392
(1997)). Total MAO activity was determined by incubating the liver
preparations for 30 minutes at 37 degrees Centigrade with a 1/1
dilution of a test fraction (compound or substance to be tested
dissolved in distilled water) or control condition (water alone).
This incubation allowed the test compound or substance to interact
with the enzyme under physiological conditions. The final tissue
concentration in the reaction mixture was 3.5 mg per 100 ml.
[0058] Following the incubation with test compounds/substances or
control, the MAO reactions were initiated and the reactions were
incubated at 37 degrees Centigrade. The reaction was initiated by
mixing 150 .mu.l of preincubated tissue with 150 .mu.l of
chromogenic solution (containing 10 mM vanillic acid, 5 mM 4-amino
antipyrene, 20 units/ml of peroxidase in 0.2 M potassium phosphate
buffer final concentration pH 7.6), 600 .mu.l of amine substrate
(tyramine 500 micromolar), and 100 .mu.l of distilled water (1 ml
reaction volume). The standard reaction time was for 1 hour, but
reaction times varied from 1 minute to 3 hours to evaluate the time
course of the reaction in the presence or absence of test substance
or control. The reactions were terminated by the addition of 30
.mu.l of a stop solution of phenelzine (10 mM). The stopped
reactions were stored on ice and placed at room temperature for
reading in a spectrophotometer at a wavelength of 498 nm. The
resulting values were analyzed to determine the amount of reaction
product produced by MAO activity. This assay was reliable and
simple to perform. A standard curve using hydrogen peroxide for
enzyme activity was prepared for each experiment to determine the
activity of the enzyme.
[0059] Selective assays of MAO A and MAO B isoforms were determined
by using selective inhibitors of each of these enzymes. During the
preincubation of the enzyme with the test solutions, either
pargyline or chlorgyline (final drug concentrations in the reaction
mixture of 500 AM) was added to the reaction mixture. This
technique allowed for the assay of MAO A or MAO B activity in the
absence of the activity of the other isoform of the enzyme. All
other reaction conditions were conducted as for total MAO activity
studies.
[0060] Each of the compounds and substances of the present
invention were evaluated by initially determining a concentration
curve at a reaction time of one hour. After determining the
concentration curves of each compound or substance on MAO activity,
a reaction time course in the presence or absence of test compound
or substance was determined and time course curves were generated.
Following these experiments, the effect of each test compound or
substance was evaluated on MAO A and MAO B activity by the same
reaction studies as described above for the total enzyme
activity.
Example 1
[0061] Anabasine, in its purified form, was dissolved in distilled
water in a maximal inhibition concentration of 0.2 mg/ml, and
tested according to the procedure described above. At maximal or
saturating inhibition concentrations, anabasine was effective at
inhibiting MAO activity by approximately 10-13%, and was effective
at inhibiting the enzyme at all time points in the reaction.
[0062] FIG. 1 presents the means (plus or minus the standard errors
of the means) for the percent inhibition of MAO activity produced
by saturating concentrations of anabasine over 60 minutes of MAO
activity measured as described above. Each data point represented
the mean of 5 determinations. All the data points shown in FIG. 1
were statistically, significantly different from the sham control
at each time point tested (student t test, p<0.01), and were
representative of multiple experiments.
[0063] Since anabasine was an inhibitor of MAO activity, further
studies were conducted to evaluate if this agent was inhibiting MAO
A or B activity using the methods described above. Anabasine was
found to inhibit both MAO A and MAO B activity as shown in FIG. 2.
FIG. 2 presents the means (plus or minus the standard errors of the
means) for 5 determinations for the percent inhibition of MAO A and
MAO B activity. The effects of anabasine on both forms of MAO
activity were statistically, significantly different from control
enzyme conditions (student t test, p<0.05). The results
demonstrate that anabasine inhibits both MAO A and B forms of the
enzyme.
Example 2
[0064] Anatabine in its purified form, was dissolved in distilled
water in a maximal inhibition concentration of 0.1 mg/ml, and
tested according to the procedure described above. At maximal or
saturating inhibition concentrations, anatabine was effective at
inhibiting MAO activity by approximately 60%. This result shows
that anatabine may be much safer as a medication than standard MAO
enzyme inhibitors. Anatabine was effective at inhibiting the enzyme
at all time points in the reaction, and was equally effective in
inhibiting both MAO A and MAO B activities.
[0065] FIG. 3 presents the means (plus or minus the standard errors
of the means) for the percent inhibition of MAO activity produced
by saturating concentrations of anatabine over 60 minutes of MAO
activity measured as described above. Each data point represented
the mean of 6 determinations. Anatabine was an effective MAO
inhibitor at maximal concentrations, inhibiting the enzyme by
approximately 60%, as discussed above. All the data points shown in
FIG. 3 were statistically, significantly different from the sham
control at each time point tested (student t test, p<0.005) and
were representative of multiple experiments.
[0066] Since anatabine was an inhibitor of MAO activity, further
studies were conducted to evaluate if this agent was inhibiting MAO
A or B activity using the methods described above. Anatabine was
found to inhibit both MAO A and MAO B activity as shown in FIG. 4.
FIG. 4 presents the means (plus or minus the standard errors of the
means) for 6 determinations for the percent inhibition of MAO A and
MAO B activity. The effects of anatabine on both forms of MAO
activity were statistically, significantly different from control
enzyme conditions (student t test, p<0.01). The results
demonstrate that anatabine inhibits both MAO A and -B forms of the
enzyme.
Example 3
[0067] Nornicotine in its purified form, was dissolved in distilled
water in a maximal inhibition concentration of 0.08 mg/ml, and
tested according to the procedure described above. At maximal or
saturating inhibition concentrations, nornicotine was effective at
inhibiting MAO activity by approximately 80 to 95%, and was
effective at inhibiting the enzyme at all time points in the
reaction. Nornicotine was also equally effective in inhibiting both
MAO A and MAO B activities.
[0068] FIG. 5 presents the means (plus or minus the standard errors
of the means) for the percent inhibition of MAO activity produced
by saturating concentrations of nornicotine over 60 minutes of MAO
activity measured as described above. Each data point represented
the mean of 6 determinations. Nomicotine was an effective MAO
inhibitor at maximal concentrations, inhibiting the enzyme by
approximately 80-95%, as discussed above. All the data points shown
in FIG. 5 were statistically, significantly different from the sham
control at each time point tested (student t test, p<0.01) and
were representative of multiple experiments.
[0069] Since nornicotine was an inhibitor of MAO activity, further
studies were conducted to evaluate if this agent was inhibiting MAO
A or B activity using the methods described above. Nornicotine was
found to inhibit both MAO A and MAO B activity as shown in FIG. 6.
FIG. 6 presents the means (plus or minus the standard errors of the
means) for 6 determinations for the percent inhibition of MAO A and
MAO B activity.
[0070] The effects of nornicotine on both forms of MAO activity
were statistically, significantly different from control enzyme
conditions (student t test, p<0.01). The results demonstrate
that nornicotine inhibits both MAO A and B forms of the enzyme.
Example 4
[0071] The Yerbamat extract was prepared as follows: Yerbamat
materials (obtained from STAR TOBACCO, INC.) were shredded and
mixed with a water/ethanol (1/1 by volume) solution in a mixture of
about four leaves per 10 ml of the water/ethanol mixture; the
materials were then extracted overnight with continuous stirring;
the solution was then removed from the Yerbamat residue and stored;
the residue was then further extracted overnight two more times
with the same volume of water/ethanol mixture, and the three
extracts were combined and filtered to remove the particulate
Yerbamat material; and the combined extracts were subjected to
removal of the water/ethanol by vacuum evaporation. The resultant
extract was then weighed and solubilized in distilled water.
[0072] When tested, Yerbamat extract was effective in inhibiting
MAO activity. The maximal inhibition concentration was 10 mg/ml. At
maximal or saturating inhibition concentrations, the Yerbamat
extract inhibited MAO activity by approximately 40 to 50%. The
results suggest that Yerbamat may be much safer as a medication
than standard MAO enzyme inhibitors. The extract was effective in
inhibiting MAO at all time points in the reaction, and was equally
effective in inhibiting both MAO A and MAO B activities.
[0073] FIG. 7 presents the means (plus or minus the standard errors
of the means) for the percent inhibition of MAO activity produced
by saturating concentrations of Yerbamat over 60 minutes of MAO
activity measured as described above. Each data point represented
the mean of 5 determinations. Yerbamat was an effective MAO
inhibitor at maximal concentrations, inhibiting the enzyme by
approximately 40-50%, as discussed above. All the data points shown
in FIG. 7 were statistically, significantly different from the sham
control at each time point tested (student t test, p<0.005) and
were representative of multiple experiments.
[0074] Since Yerbamat was an inhibitor of MAO activity, further
studies were conducted to evaluate if this agent was inhibiting MAO
A or B activity using the methods described above: Yerbamat was
found to inhibit both MAO A and MAO B activity as shown in FIG. 8.
FIG. 8 presents the means (plus or minus the standard errors of the
means) for 5 determinations for the percent inhibition of MAO A and
MAO B activity.
[0075] The effects of Yerbamat on both forms of MAO activity were
statistically, significantly different from control enzyme
conditions ( student t test, p<0.01). The results demonstrate
that Yerbamat inhibits both MAO A and B forms of the enzyme.
Example 5
[0076] The tobacco extract was prepared in the same manner as in
Example 4, except that processed tobacco leaves (obtained from STAR
TOBACCO, INC.) were substituted for the Yerbamat materials.
[0077] When tested, the tobacco extract was effective in inhibiting
MAO activity. At maximal or saturating inhibition concentrations,
the tobacco extract was able to inhibit MAO activity by
approximately 60%. The results suggest that the extract may be much
safer as a medication than standard MAO enzyme inhibitors. The
tobacco extract was effective at inhibiting MAO at all time points
in the reaction, and was equally effective in inhibiting both MAO A
and MAO B activities.
[0078] FIG. 9 presents the means (plus or minus the standard errors
of the means) for the percent inhibition of MAO activity produced
by saturating concentrations of tobacco extract over 60 minutes of
MAO activity measured as described above. Each data point
represented the mean of 8 determinations. Tobacco extract was an
effective MAO inhibitor at maximal concentrations, inhibiting the
enzyme by approximately 60%, as described above. All the data
points shown in FIG. 9 were statistically, significantly different
from the sham control at each time point tested (student t test,
p<0.001) and were representative of multiple experiments.
[0079] Since tobacco extract was an inhibitor of MAO activity,
further studies were conducted to evaluate if this agent was
inhibiting MAO A or B activity using the methods described above.
Tobacco extract was found to inhibit both MAO A and MAO B activity
as shown in FIG. 10. FIG. 10 presents the means (plus or minus the
standard errors of the means) for 8 determinations for the percent
inhibition of MAO A and MAO B activity. The effects of tobacco
extract on both forms of MAO activity were statistically,
significantly different from control enzyme conditions (student t
test, p<0.005). The results demonstrate that tobacco extract
inhibits both MAO A and B forms of the enzyme.
Examples 6 and 7
[0080] The extract of GUMSMOKE chewing gum or lozenges was prepared
as follows: five slices each of gum or lozenges, formulated with
tobacco extract, were extracted with 50 ml of distilled water at
room temperature for 12 hours. The undissolved gum substance was
removed by filtration. (The lozenges dissolved completely.)
Dilutions of these extracts were prepared for evaluation.
[0081] The gum and lozenges extracts were effective in inhibiting
MAO activity. At maximal or saturating concentrations, the extracts
were able to inhibit MAO activity by approximately 50 to 60%.
[0082] FIG. 11 presents the means (plus or minus the standard
errors of the means) for the percent inhibition of MAO activity
produced by saturating concentrations of an extract of GUMSMOKE
chewing gum prepared as described above over 60 minutes of MAO
activity measured as described above. Each data point represented
the mean of 4 determinations. GUMSMOKE extract was an effective MAO
inhibitor at maximal concentrations, inhibiting the enzyme by
approximately 50-60%. All the data points shown in FIG. 11 were
statistically, significantly different from the sham control at
each time point tested (student t test, p<0.05) and were
representative of multiple experiments.
[0083] FIG. 12 presents the means (plus or minus the standard
errors of the means) for the percent inhibition of MAO activity
produced by saturating concentrations of an extract of the lozenge
prepared as described above over 60 minutes of MAO activity
measured as described above. Each data point represented the mean
of 4 determinations.
[0084] The lozenge extract was an effective MAO inhibitor at
maximal concentrations, inhibiting the enzyme by approximately
50-60%. All the data points shown in FIG. 12 were statistically,
significantly different from the sham control at each time point
tested (student t test, p<0.05) and were representative of
multiple experiments. Both MAO A and MAO B were also inhibited by
these extracts.
* * * * *