(12) STANDARD PATENT APPLICATION (11) Application No. AU 2014202047 A1 (19) AUSTRALIAN PATENT OFFICE
(54) Title Methods for treating dependence
(51) International Patent Classification(s) A61K 31/415 (2006.01) A61P 25/30 (2006.01) A01N 43/50 (2006.01)
(21) Application No: 2014202047 (22) Date of Filing: 2014.04.11
(43) Publication Date: 2014.05.01 (43) Publication Journal Date: 2014.05.01
(62) Divisional of: 2008283903
(71) Applicant(s) Biotie Therapies, Inc.
(72) Inventor(s) Pickford, Lesley;Woiwode, Tom;Moran, Mark
(74) Agent / Attorney Spruson & Ferguson, L 35 St Martins Tower 31 Market St, Sydney, NSW, 2000 2014202047 11 Apr 2014 phase dependence Compound Provided symptom
of
cocaine
are of
A.
on
abuse methods
Also
at
dependence
least
of,
provided
dependence of
one
treating
substance
in are
patients.
patients methods
on,
in or
patients
ABSTRACT
withdrawal
suffering of
treating
and
from
certain from at
least
or
at
susceptible
methods
least one
phase
one
of
substance
to of
treating
at substance
least
at with
one
least
one
2014202047 11 Apr 2014 U.S. nationwide, which troublesome, their phase herein Dependence Dependence Drug illicit In Background of Field Treating Cross-Reference current approximately legal substance despite and substance at [001] [004] [003] [002]
treating 2004, least
use
lifetimes
of Provisional drug
Abuse
concern. of is by
Provided This The Substance
significant
crack
Invention
with
one greater Dependence substance
approximately reference
with (alcohol at abuse
and problem
of application Using
least Using
Warning symptom in loss
users
and
13.9% the Compound
cocaine than
2005,
to are alone According
abuse
one Patent
of
substance Invention
METHODS
3.3% or Nepicastat Related dependence in
Nepicastat increased
certain
in of control
Using of their
illicit phase
is Network there
of 2004 was and
cocaine
22.5 claims Americans have $181 Application
A. abuse entirety.
Applications
Nepicastat methods
drug) of dependence the to were use in
when
million ” from ” tried billion
Also
cocaine FOR
filed on the limiting the involved
estimated filed abuse related
of,
abuse.
2.4 at
the
benefit crack aged
467,000
provided
2005
August TREATING
of
least dependence
(2002). Americans
August
”
Serial million number dependence and
treating filed intake problems
are
in 12
cocaine The
National one
940,953
of
the dependence
characterized and 6,
in
October Nos. are
and 17, persons
was substance latest of 2007, majority 2004
patients
older
aged
certain DEPENDENCE
the
on, and at 2007,
priority
in 2.0 60/935,323, drug-related
Survey
least estimate
60/956,555, to patients. 4,
substance.
have or at million. 12 who
2007, of 682,000 is
suffering methods the in and
withdrawal by
or once these.
under a
tried patients
were
on expense major older substance
60/960,591, for which
in
Similarly,
Drug “
emergency cocaine
in Methods These 35
of the “ current
from their Methods
needed
medical,
2005. and
treating
are U.S.C. of
costs from
Use
craving,
or
lifetimes. other behaviors
certain all
at
cocaine
the
“ susceptible
treatment
to
Methods for In least incorporated at
and for room § at
social,
number behaviors. society
2004, least
119(e)
least
methods Treating Treating seeking,
once
Health,
users, occur More visits
one one and the
for for
of of to to in
2014202047 11 Apr 2014 bromocriptine, variety treatments plasma metabolism necessary pharmacologic those well. particular, including occur increased intranasal in of including completed, Clinical completed, dosing alterations consists esterases and disulfiram discouraging. label acting agents, [005] [007] [006]
vigabatrin elevations
plasma
for trials. in None
on Inhibition Clearly, The of documented cocaine including of
trials
the baclofen
tiagabine,
and treatments cofactor plasma GABA a in non-specifically numerous flushing,
cocaine such
as of
dopamine including
esterases.
of
medication physiologic context
have
of
in alcohol have
pergolide, The DBH.
treatment levels these there
as
mood
plasma of ritanserin,
have systems cocaine
for been
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outcomes
that plasma
have
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compounds were is (ethanol),
a trials β
stabilizers,
been
By Disulfiram
-hydroxylase uncontrolled
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or a disulfiram component amantadine,
cocaine
for of
been
levels. have
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need observed treatment
subjective and esterases inhibiting
gepirone,
phenytoin, slightly of behavioral
and cocaine
from
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of producing been desipramine,
equally
for controlled
vigabatrin. has
levels. including enzymes,
several and Increased
slowed
more in
these a
may illicit slows
mazindole, (DBH) proved effects with nimodipine, evaluated
dependence
aldehyde in 2
one broadly
related direct therapy
encouraging,
clinical
encouraging, the
be
enzymes, In
use. the controlled
development the
disulfiram,
clinical carbamazepine
including of
inhibitor Results reliably cocaine disulfiram-ethanol fluoxetine,
or more
laboratory
elimination
cocaine,
compounds elimination
effective or as
and A dehydrogenase, indirect trials,
and
currently
contingency treatments
subsequent
trials
including methylphenidate.
effective
for
efficacious.
study, though levels aldehyde
disulfiram
naltrexone
though without disulfiram however.
tiagabine
treatment studies efforts dopamine of bupropion,
of
chelate and of
and
available. antidepressants were
based
cocaine,
for than aldehyde not
cocaine,
study noteworthy
management
dehydrogenase, lithium
reaction.
greater
have
evaluating disulfiram is
have
Six-fold have cocaine
compelling.
treatment Several not
copper, approach, primarily agonists, current the
using
and
which A
been
associated been generally presumably have elevations dehydrogenase
most Unfortunately, range
This elevations
dependence, imipramine. medications
success.
IV which
behavioral have equivocal, studied
effects alters
alone. can also markedly including on
effective
and reaction of cocaine Studies plasma
open
result
other
been been been
with may
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by
of In as in A a
2014202047 11 Apr 2014 positive the relative proportion mg/day respectively. Americans, with norepinephrine this positive inhibiting of DBH In cocaine cocaine cocaine 62.5 corresponding fairly accounted and synaptic associated disulfiram, compared suggested Similarly, [008] [010] [009]
cocaine-positive
increases this
outcome
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mg
diminished activity. common.
Disulfiram Several Several
use dependence dependence. to relative urine concentration subjective vesicles study, and
to
plasma
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22% during and
placebo, remains in
125 cocaine-positive by samples
haplotype It (NE).
reports studies plasma
disulfiram
to
reduced 112
DBH
provided among variability The and has
mg esterases.
treatment inhibits placebo
effects (p=0.001).
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cocaine-dependent is disulfiram/day frequency DBH
of concentrations compared have indicate gene
be
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frequencies
replicated. 250
shown
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shown transcription with Slow
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of of
along
to
that use
disulfiram.
European DBH outcomes alcohol highly
absorption the
the preliminary per
placebo two
disulfiram
(p's increased
by
observed
opiate- over in documented
are the
with
single 3
T
locus.
day in
cocaine.
subjects
<
psychotherapies.
volunteers
heritable and use allele
studies,
efficacy 0.32, noradrenergic 0.04).
time
when
Americans NE. treatment.
was increased
Recently, pattern following
and
with
enzyme over
earlier. is The
efficacy
is
0.34,
during with
more DBH associated
treated
treatment In
reported
Patients
by cocaine-dependent of and
time lower T
were
or
those
variant
250 low
the and
over
that
and variability results effective The can type
intranasal
of treatment
with neurons
with
DBH
provision mg/day DBH
with randomized
disulfiram 0.09 with
to be
with 16% time effect mediates Disulfiram of
with
(-1021C
disulfiram
disulfiram, be from
measured
psychotherapy
activity.
high activity,
for in comorbid among disulfiram and
during
20%
dosing reduced in size with
disulfiram
patients
a
these of
is — activity DBH large as
the patients
>T) was
among localized
fewer to
disulfiram at Japanese. treatment
treatment a the in
up accounted
This
is synthesis
62.5 treatment
populations, cocaine
clinical alcohol activity, reduced modest the placebo
with
to
proportion
associated
is provided.
treatment
African-
cocaine 500 allele reduced plasma, mg/day
largely
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250
and and use the
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2014202047 11 Apr 2014 those possibly therapeutically the treatments methods inhibits Description one DBH on genotype abuse-related genotype. appears extinction, a acquisition, Compound cocaine effective Summary [Oil] [017] [016] [015] [014] [013] [012]
patient,
at
patient symptom
is with least
Also Also Disulfiram Fig. Fig. Provided While
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DBH
by
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include key associated of
in
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A.
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observation that
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sizes in
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appear from phase
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assays. from A. patients
esterases,
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confirms
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is
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2014202047 11 Apr 2014 treatment mixtures vehicle with vehicle varying or hydrochloride Detailed a (5,7-difluoro-l,2,3,4-tetrahydronaphth-2-yl)-2,3-dihydro-2-thioxo-lH-imidazole, l,2,3,4-tetrahydronaphth-2-yl)-2,3-dihydro-2-thioxo-lH-imidazole, context generally dosed [021] [020] [019] [018] [028] [027] [026] [025] [024] [023] [022]
range
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Description
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2014202047 11 Apr 2014 pharmaceutically that to the that base by p-toluenesulfonate, maleate, inorganic hydrochloride). one obtained sulfinate, l,2,3,4-tetrahydronaphth-2-yl)-2,3-dihydro-2-thioxo-lH-imidazole, difluoro-l,2,3,4-tetrahydronaphth-2-yl)-2,3-dihydro-2-thioxo-lH-imidazole alleviating, acid, an such affected [030] [029] [036] [035] [034] [033] [032] [031]
either
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term
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compounds. as
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As In “ The The The The The
acetate,
accordance Pharmaceutically by directly "patient"
be by
nitrate, addition, acids, fumarate,
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particularly
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Conversely,
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for acid "administration,"
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hydrochloride
(R)-5-Aminomethyl-l-(5,7-difluoro-
to
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product salt, acid,
limited certain salt.
condition,
as the likelihood
the to or addition methanesulfonate, as
used may
a
more
solution free such
patient.
methodologies
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be
herein,
base
well a
completely
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salts salts
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with malate, sulfate,
at
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2014202047 11 Apr 2014 the the with with the problems; related pattern involves irritability, includes interpersonal occurring impairment is of criteria encompass develops substance embodiments Compound substance least embodiments, criteria situations fulfill Text [038] [037] [039]
the defined
DSM-IV frequency
substance
revision nepicastat Compound one
major American
As to The Those of for set
by
certain symptoms
DSM-IV the
within
and being in used of substance
treatment A as recklessness, substance forth
term
or
way
role problems repeated (2000) the reduces which
TR
abuse of maladaptive (4) treatment
and of reduces herein,
distress,
A
symptoms Psychiatric
a patient abused, in
ordinary "substance
of obligations requires continued
12-month TR abuse,
in
amplitude the
of ("DSM-IV
it is of
use example
the
dependence. a use
caused the symptom
is substance physical
the withdrawn
Diagnostic patient
enters
poor of
as manifested substance
and
physically of
skill phrase known that
pattern including
amount substance
substance Association. manifested
period: substances.
abuse,
at judgment,
mood or and
remission of
reduces
TR"), the in
work, exacerbated
for and/or
abuse
“ a
the
reduces without as
of ” craving In and from or
symptoms
symptom
substance by disorders.
sleep
as (1)
hazardous;
which
use abuse some substance frequency "withdrawal.
art
school,
7 itself, at Statistical
recurrent psychological used by and compulsion, recurrent As
a A
despite
least
will
a and induced
limitation
dependent
was embodiments,
at feature recited with no symptom by
dependence,
herein,
of
abuse. of or
mood
appreciate least one
longer the
prepared In
of abuse
having a and Compound home; substance Manual substance (3)
”
by effects some substance in of
condition symptom
one
disturbance aggression,
” can
at
the
a significant recurrent experiences
substance In “dependence. individual, The refers leading
stressful
least persistent
(2)
by
some
embodiments, DSM-IV be of and that treatment of of
use methods the abuse
recurrent
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of use
one in also
“
embodiments,
to
substance reducing Task
in resulting
substance-related
event substance substance. abuse
and a anger, adverse
in following
the of the or of clinically have TR, a patient.
”
of
a
withdrawal. patient recurrent
Force Disorders, intense
with
euphoria, symptom.
described in patient. substance
individual is
Also,
substance substance
craving treatment
a never
at a in consequences
patient.
abuse
abuse
reference maladaptive on
least In
a
symptoms, reduces
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when treatment
failure
DSM-IV addition, social In
met
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4 for certain
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th abuse abuse
some often often legal with
Ed.
the the the
of or of to to in at
2014202047 11 Apr 2014 reduces need recklessness, treatment physiological persistent unsuccessful taken DSM-IV of of in or of important in any distress criteria embodiments, least depended substance some dependence, caused amounts syndrome substance; (b) [041] [040] [042]
a the
substance activities frequency
example substantially patient time
for
one embodiments, to frequency
As As or The
set
the
as relieve
substantially
or
within TR
upon, exacerbated
dependence social, or of for used symptom
used forth
manifested (2)
reduces
term substance
over
to
use; efforts is
recurrent poor and where substance dependence, the the of
withdrawal,
obtain
a
herein,
or and
in
herein,
the and
"substance
occupational pattern a diminished substance term and
specific the avoid without judgment,
at
longer
to
Compound mood same amplitude of
craving increased evidence
least by DSM-IV (7) the in cut by remission the
physical “
substance
dependence withdrawal of remission
the a
the twelve at down
disorders. substance,
phrase where period substance; one patient.
as substance dependence,
limitation
use least
substance. induced
substance effect compulsion,
or demonstrated
of
symptom A amounts TR.
of or
or
by does
recreational month a no “ treatment than three
”
increases
control
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In tolerance psychological
refers symptoms; The with
a
evidence In
use
reduces by use,
or
not
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8 some ” selected use
Substance of
period: of
(b) a symptoms
as
the
or apply
leading continued substance stressful least
to
aggression, the
substance
of
intended; used is a
the embodiments, by of
dependence
embodiments, substance,
activities a symptom
at of substance continued substance or
a
if (3) state
either
from same, one
In (1) herein, condition tolerance least
the
to
withdrawal dependence problem
event the for
some use;
tolerance
clinically during
dependence use patient
of
(4) the
substance (a) ” anger, substance or one are
can
or
dependence to
in despite
(5) refers
euphoria, there has of following a
the embodiments, in
or treatment given
achieve recover
a treatment
that DSM-IV
closely be which is a
patient. the a
withdrawal craving
been great as
can characteristic significant on patient. defined is to
is
knowledge is
dependence
which is up defined same agonist increasing
a
be
present,
the the
likely often
from deal related
reduced.
persistent apathy, group,
reduces or
with
with with TR
for
desired with
occurrence reduced
includes amount
of
taken by its therapy impairment is Compound
the to Compound
symptom Compound physiological
time
substance occurring
reference present. of or
withdrawal either at the effects;
set irritability, have
substance
effect; desire least having
In in
because without is amount forth by
of
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for (6)
In or or or to in A A A at at is a a a
2014202047 11 Apr 2014 remission which remission patient. been than been remission remission, have In of symptom embodiments controlled criteria least least substance less at dependence dependence [043] [048] [047] [046] [045] [044]
least some remission
than
twelve met, met. 12 been one
no In In The The The The
one for
months, embodiments,
twelve
symptom but refers dependence some refers of some
symptoms reduced
environment substance and
symptom term qualifier early qualifier months. (early have term
substance are the remission
only embodiments,
to
embodiments, to
"sustained at months, criteria based been
"sustained partial
versus a
in a least of
state
"early
dependence state "early of of
a applies
or
present Compound substance
patient.
substance abuse on
where
substance
refers
for substance one sustained during no remission, during
partial the partial
full
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symptom full symptom
after the
or for Compound
access to interval
remission" In which
or
which dependence
dependence dependence remission" remission" remission at A
a remission"
abuse remission) some
abuse
substance
none
state treatment least
sustained
dependence
to substance of of
9 of
all
embodiments,
have
or the
one substance substance during
of
A time is symptoms
is dependence
is
treatment
relevant is abuse is used or the
or and
month. does been prolongs
characterized
used
used
that
used abuse full
substance use
which
whether
or symptoms
when
met
have not
when dependence
dependence does substance
has when
remission, of when
substance The (partial
remission promotes
a at occur.
the
occur. substance not
for elapsed period
not
for there
any definition for abuse
by been occurrence none
at
occur.
at for
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time
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abuse
least
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In
or or In is least remission
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and has since met. substance continued least
of
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one
full
have of one
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of
remission).
month, or month
embodiments, embodiments, not a symptoms
a
met,
in
cessation at of state dependence period presence
period abuse months,
four been abuse abuse
a least early In
but patient.
but but partial during
types some
met. of
in less and one
full has has
the for
of of of
at at a
2014202047 11 Apr 2014 A remission remission. time Amphetamine-Induced Amphetamine-Induced Amphetamine-Induced Alcohol-Induced Alcohol withdrawal withdrawal Psychotic Intoxication; Dysfunction; Induced Induced Withdrawal; one In or induced Withdrawal sustained cause compulsion, example some general a Specified [050] [049] [051]
relevant
treatment other some
of during
remission embodiments,
The The "Withdrawal"
the medical
by
important
Persisting Dependence;
Mood partial
embodiments, and will substance
Disorder,
(NOS);
a can
which term
phrase
In following in
usually,
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end
some
Amphetamine without a
cause to remission,
condition Disorder; substance
patient.
Psychotic
the
in areas end “
Amphetamine
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With but Withdrawal embodiments,
refers treatment
a event
patient clinically
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conditions at patient Alcohol Sleep in
Psychotic
Anxiety limitation of
anger, treatment
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a
and
and
Hallucinations;
to in dependence functioning, Alcohol-Induced In
patient.
a Disorder, is
necessarily, Disorder;
a a
after
Sleep
period
are some sustained one delayed, in Alcohol-Induced patient. substance significant Intoxication with collection
Intoxication;
Delirium;
remission.
characterized Disorder;
not
Abuse; with Disorder, of a apathy, Compound
In
Disorder;
Compound embodiments, stressful
of
early
better 10
some Amphetamine-Related
With can or
for
full
Compound remission
is
of craving,
distress stopped.
Amphetamine-Induced
Alcohol-Induced
example. Amphetamine be
associated partial remission.
symptoms irritability,
In accounted embodiments,
Amphetamine-Induced Persisting
With Hallucinations; Alcohol event. Delirium;
Alcohol-Related
characterized A some
in
Psychotic A
treatment
or ” mood
the
remission,
The
withdrawal reduces refers
Delusions; A impairment embodiments, In
These
that
with DSM-IV Intoxication
for reduces some substance-specific recklessness, Amnestic disorders,
Amphetamine
to Dependence;
arise
by
relapse
Disorder, Anxiety reduces substance symptoms by
at
increasing
embodiments, Disorder Alcohol-Induced
sustained
another
Disorder
the least
when symptoms the Amphetamine-Induced in TR:
Sexual
occurs and a
Disorder; social, presence
Delirium;
the Disorder;
substance
stressful
one Mood
administration With
dependence. mental poor Alcohol are NOS;
sleep
Not likelihood the full
Amphetamine
Dysfunction; at symptoms occupational symptom not Withdrawal;
include
Compound
interval Delusions; the
remission, judgment, of
Otherwise
disorders.
event Disorder;
Cannabis
Alcohol- Alcohol- disorder.
due Alcohol
craving at Abuse; Sexual end
least
to that
can
for
of of of of of In a
2014202047 11 Apr 2014 Nicotine Nicotine-Related NOS; Abuse; Abuse; physiological physiological the Dysfunction; Intoxication Disorder Delusions; Remission; Withdrawal; Remission; Phencyclidine-Related Delusions; Phencyclidine-Induced Phencyclidine Induced With Induced Persisting Inhalant-Induced Cocaine-Induced Cocaine-Induced Cannabis-Induced Opioid Opioid Opioid Opioid Sustained [052]
following
Delusions;
Phencyclidine The Withdrawal; Dependence, Abuse;
Dependence, Cannabis
Inhalant
Sexual
Psychotic
NOS; Dependence,
Dementia;
Cannabis-Induced Partial
terms
Opioid Nicotine
Phencyclidine-Induced Opioid-Related
Cocaine-Induced
Delirium;
dependence; dependence;
conditions
Opioid
Intoxication Dysfunction; Cocaine
Mood
Anxiety
Dependence; Dependence; "cessation Psychotic Opioid-Induced Disorder
Remission;
Anxiety
Opioid-Induced Dependence Disorder,
Early
Sustained
Abuse; Inhalant-Induced Dependence,
Disorder
Mood Dependence; Disorder;
Early
Cocaine Abuse;
Disorder;
Full
characterized
” Nicotine
Disorder
Opioid
Not Disorder, Psychotic Disorder;
Disorder;
and
Opioid-Induced
Phencyclidine Delirium; Sleep
With Remission; Full NOS. Nicotine Cannabis Inhalant Full
On Cocaine Cocaine-Induced “
otherwise
withdrawal"
Psychotic
Withdrawal;
Mood
Sustained Inhalant-Induced
Opioid
Disorder; Psychotic Remission; Dependence, Remission; Not Hallucinations; Agonist Dependence,
11 Disorder, Phencyclidine-Induced
With Psychotic
Cannabis-Induced
in Intoxication;
Otherwise Intoxication;
Dependence,
Disorder; Phencyclidine-Induced Dependence; Opioid
Intoxication; the
Dependence;
Hallucinations; Disorder, Specified;
Sleep
Therapy; Cocaine-Related Full
may
Disorder, DSM-IV Opioid With
Nicotine Cocaine-Induced Dependence,
Disorder,
Psychotic without
without Specified;
Opioid-Induced be, Remission; Disorder;
Mood
Inhalant-Related
Hallucinations; Cannabis Inhalant
On but
Cocaine Dependence, and With
Opioid
Nicotine
TR:
Dependence, Phencyclidine With Psychotic need
physiological
Disorder; physiological Agonist
With Disorder, Opioid
Psychotic Opioid Cocaine-Induced
Opioid-Related
Hallucinations; Early Disorder
Intoxication Nicotine
Intoxication Intoxication Nicotine not
Intoxication;
Hallucinations; Delusions;
Anxiety Psychotic Anxiety Dependence,
be,
Partial
Dependence Dependence,
Cannabis-Related Therapy;
Sustained Inhalant-Induced With Disorder, Disorder
Disorder, NOS; in
Early Dependence,
Intoxication; Withdrawal; dependence; dependence;
reference
Remission;
Delusions;
Delirium; Delirium; Delirium;
Disorder; Disorder; Disorder,
Inhalant-
Disorder
Inhalant Cocaine Opioid-
Opioid Sexual
Partial Partial
NOS;
With With with with in and
to a
2014202047 11 Apr 2014 nicotine those better withdrawal restlessness; by nicotine the the typically withdrawn tobacco tobacco Environment; Withdrawal, Withdrawal, Dependence; impairment irritability, in For Dependence including, Ethanol substance controlled eliminate described least complex [053] [054]
any
symptoms oral symptoms example,
one
who accounted form, As Withdrawal
use
or includes,
replacement or
results organic Withdrawal,
response
entirely
herein
abuse, used
but environment;
intranasal have when
in
frustration from nicotine. or
On Early Sustained decreased including,
Amphetamine
of the social, reduction Ethanol not
Agonist
including herein, in
nicotine
for
may
replaced, compound their such
dependence, discontinued
but
by Full
or the symptoms limited
by
occupational,
therapy. partial ingestion
be
is the Full dependence
onset
symptoms Remission; on heart but These
“ another or
Therapy
on in Withdrawal, Ethanol not withdrawal used
not Remission;
Agonist or dysphoric, patient. the
anger; to
agonist Withdrawal;
response or rate; of
limited
can
partially symptoms
to a
limited amount or of Thus, nicotine use a medical
”
chemical
protein,
alleviate are Withdrawal;
increased
refers arise tobacco
withdrawal. Ethanol on
or
anxiety; of therapy
Therapy; as
not to other nicotine of
such For
to
Ethanol
tobacco 12 of
upon without
depressed a
replaced,
to withdrawal disorder.
snuff
the
and consequence due smoking
often nicotine
that
one Opioid Withdrawal, or
compound, example, important patients ”
appetite
substance
nervous
reduction Cocaine refers
to
chewing
in
triggers and
Withdrawal, Ethanol or Ethanol
products,
cause a
The
all
Physiological
their more Dependence general use, of The
mood;
chewing
to forms.
can conditions.
or
cigarette, areas
term Withdrawal. in
tremor;
of being
being
tobacco.
is of a present clinically use
such weight Early
Dependence symptoms
be
Withdrawal, the response
often some all any medical
of light-headedness;
“ assisted Sustained of
tobacco. agonist discontinued treated of abused, as
Partial functioning. of
on method tobacco gain; cigar, followed
difficulty
which
a embodiments, Individuals a Such Dependence;
significant
methadone
in variety condition small attributed
”
to
with
Remission; and
with or
depended a refers
Partial
In oral The is reduce
contain patient
with pipe
within
the
molecule
also use an concentrating; of
a Physiological
The
or
cessation
often
to and tobacco, Remission; substances. craving agonist therapy. to distress the
Controlled
of
helpful insomnia; intranasal
and
that
24 nicotine, upon, methods a
nicotine “ tobacco Ethanol Ethanol
are Opioid
use
factor suffer hours
even or is
not
for for
or of or or of to at a
2014202047 11 Apr 2014 may that results medical psychomotor nausea; reduced. beverages) medical not withdrawal naloxone opioids; hyperactivity or lacrimation; condition administration, sociability; attention symptoms. symptoms areas ethanol conditions alleviate symptoms longer-acting fever; clinically [055] [057] [056]
other
due are
include
of
and
in The Cocaine The
condition decline disorder. irritability functioning. generally
one important vomiting; to
deficit or
the
significant These can
often
attributed
are results
insomnia. less symptoms a naltrexone Symptoms discontinuing discontinuing
rhinorrhoea;
or anxiety;
opioids, agitation; (such presence general
also
through
more characterized abuse
cause hunger sharply, hyperactivity ethanol
and
opposite
in and areas as be
transient
to
The
distress symptoms
restlessness;
the
such and are
clinically
medical
sweating
usually increased after
of of ethanol precipitated injection When anxiety;
and
of withdrawal methods within
not onset papillary cocaine a
dependence or of
as
to opioid functioning. characteristic
fatigue;
better
methadone, or by reduction or visual,
disorder the dependence
withdrawal occur
condition
of attributed significant or
and or impairment 4 sensitivity reduction
symptoms
abuse described muscle
opioid
use.
pulse to ethanol orally, accounted dilation;
by
a grand symptoms within
tactile,
can 12
marked
13 and administration
Opioid
in
and
rate
symptoms
The hours agonist opioid and to
aches, through
cause
when
distress withdrawal to is mal administration in euphoria;
herein opioid
6-24 in piloerection;
dependence that
greater pain; for
on or methods are
use
withdrawal feeling
social,
seizures. include
after cognitive,
such
withdrawal often
by
effects. begin short-acting auditory hours
smoking not
or may of
withdrawal dysphoric
may
another
than
increased impairment
symptoms
ethanol
ethanol
of in better of occupational,
conditions. described be
craving when take after
can
the physical an
sweating; These 100); of behavioral, is
used These or hallucinations
medical
condition.
characterized
include back 2-4 opioid mood;
accounted
an the
use intranasal opioids,
when (e.g., blood
energy, to
hand are
for
days symptoms
opioid, in withdrawal herein
last has alleviate and and
not Ethanol
social, nausea
ethanol; disorder.
diarrhea; antagonist such ethanol or varying
tremor; concentrations to
and
dose,
been
legs; mental This
such due excitement,
other
ingestion, for can emerge.
typically
symptoms
physiological by
or
occupational
or
one
to
often
withdrawal withdrawal craving by while stopped
be
autonomic containing degrees as symptoms symptoms
insomnia;
vomiting; important yawning; a illusions;
strength;
or such used another
general heroin,
These
cause more often
with self
and
are
for of or of to as
2014202047 11 Apr 2014 nasal vomiting, variance treatment treatment phase maintenance, treatment of include irritability, include irritability, abuse some cardiomyopathy, cardiovascular dysphoria; administration a substance acquisition dependence acquisition substance development acquired extreme at [060] [059] [058]
patient
least cocaine
irritation, embodiments, and of
‘ Cocaine Substance one
‘
by
Maintenance
during symptoms
suspicion, in
by substance reduces with
headache,
of being
use dependence through
phase recklessness,
decreased negative way phase at sleep
in the of a extinction,
Compound least to by
nasal the patient. withdrawal
the effects
a abused
of acquisition reduces
and at or
patient. snorting a dependence disorders, reduces
acquisition
acquisition dependence vertigo, least one and subjective example
such use Compound ”
patient. crusting,
sensation
heart refers in
or
include
of poor and
of one craving at a In
A
as depended
patient.
or symptoms
the
least at the anxiety,
reduces
phase attacks. to some relapse. In symptom
and judgment,
increased sniffing
bronchitis,
recurrent phase.
symptom
In
least phase
a
amount substance of can
in
A one some
phase symptoms for
some
without
which reduces pain; embodiments, treatment
In
the
dizziness,
upon, symptom one
be
in
cocaine. can
As can
of some Symptoms
14
embodiments, of
and
nosebleeds, substance of a
embodiments,
compulsion,
appetite,
and
cocaine characterized
used
DSM-IV include patient. dependence result
substance by shortness
cocaine limitation and
at
frequency
embodiments,
such
reduces
a craving
least of psychosis,
herein, mood
patient. in In
substance withdrawal.
a craving
Compound
also
ear,
abuse In
pyschomotor
fatigue, as one nasal some symptom
dependence
of
at aggression, some disorders. at
for
the on Compound nose,
of
heart Compound
include by
of
least least breath,
and
In and
stuffiness, induced the substance cocaine. term
embodiments,
nepicstat
abuse the lack embodiments,
some
the
and
dependence.
confusion. substance one A one palpitations,
of
amount
“
of dilated
in
acquisition throat
In
phases: and treatment retardation, and
anger,
substance
by embodiments,
of A symptom
pleasure, which
A
some use treatment Respiratory and
a
dependence euphoria, treatment
treatment chest
stressful effects
or
pupils,
is craving Administration by
facial
Compound
there embodiments,
Compound
frequency initiated
”
inhibits
a acquisition, arrhythmia,
depression, of abuse
refers pain,
patient including agitation, increases
pain.
is event
cocaine nausea, apathy, for
of a effects of which stable
10%
to
and and and
the the the the
of In
in A A is a
2014202047 11 Apr 2014 judgment, reduces without maintenance with training phase. which the treatment not maintenance reduces one In of or occurs induced irritability, depended a considered dependence Compound substance abuse development extinction extinction embodiments, [062] [061]
patient.
some exposure relapse
extinction provided symptom
substance
and
includes at
“ ‘ In
‘ prior by at Relapse Extinction embodiments, the
limitation
the
upon,
some of being dependence phase.
least in compulsion, recklessness, a phase.
A
a In
after to
phase.
phase
substance stressful end the
of
to
a of stable to
treatment
phase
use. some the an by
embodiments,
patient. and ” one relapse. the
abused
substance
a extinction of
a
refers In way environment
”
reduces
patient period
dependence remission. at
mood In extinction refers DSM-IV behavior. reduces In some
embodiments,
event
in
some
of Compound
least
craving
or aggression, reduces to poor some a In In
example
disorders. of patient.
to
abuse
depended embodiments,
or
at in
some recurrence phase some
embodiments,
one abstinence a
at
least symptom treatment judgment, a a
phase
phase
related In
embodiments, patient In least
induced on patient the
and
embodiments,
some
of embodiments, reduces A and
one In some
Compound
the anger, withdrawal
treatment upon, In
in dependence of one some euphoria,
without cue
in
of 15 of
some of embodiments, with a
substance in abstains
substance
compulsion, by the at
embodiments,
patient. the symptom
at
substance
Compound at or a and embodiments, craving
least
patient. Compound
extinction a least embodiments, least amount stimulation
limitation
of
Compound
stressful mood relapse
A
symptoms apathy,
Compound one
which from
the
dependence one In
treatment one is of
for
abuse
maintenance withdrawal
and In some aggression,
disorders.
A a
extinguished substance
use
symptom phase.
occurs DSM-IV
patient includes
the A Compound treatment some at event
irritability,
that Compound frequency
reduces
in least and A of treatment embodiments,
substance A
of
a was
after embodiments, in the
treatment has
in treatment dependence
patient
one
the
by
symptom In symptom which of abuse
anger,
phase reduces previously the a
undergone substance.
drug
way
some of A of substance or maintenance
recklessness, patient A with
substance
euphoria, being substance treatment
in treatment
and
the reduced
reduces of priming, craving
of
promotes embodiments, the
Compound the Compound
occurs
of
example
in substance
dependence
the
the
during
abused associated
extinction extinction frequency substance a abuse In
patient. craving
apathy,
relapse at relapse
for
use in during
of stress, in phase
some
poor least
and
the the the the the the
by
or or A A is
2014202047 11 Apr 2014 judgment, patient. phase pemoline, the psychostimulant psychostimulant reached while preferred methods reduces hypnotic, one stressful embodiments, depended and least dextroamphetamine, abuse Compound embodiments, Compound desired embodiments, embodiments, stages. agent [065] [064] [063]
symptom
without
one
minimizing beta-phenylisopropylamine reduces and
is Provided In
Treatment
in
therapeutic the event In
In symptom
include
before upon, mazindol, some a
selected compulsion, an a A. A
some some
patient. withdrawal medication. limitation
of
is the an treatment in the at In anxiolytic,
abuse
agent agent,
are embodiments,
escalated and
a
initial least some symptoms, induction embodiments, of
administering embodiments, amount dose patient
of
methods
and from (-) response substance
mood substance
is of,
one
an embodiments, at
of cathione, low
aggression,
with methamphetamine. a
symptoms
dependence
during opioid,
of least beta-phenylisopropylamine In in Compound
DSM-IV ecstasy, and
disorders.
of dose
side of
Compound order some Compound
is
abuse,
abuse Compound treatment treating one
Compound
the to
an an and effects, of a observed.
hallucinogen,
to embodiments,
the relapse anger, derivative initial Compound of in
phenmetrazine, symptom
illicit
the
and fenfluramine. on, dependence, determine
A
16
euphoria, In a
a
patient
A
and at is A or
patient dependence
patient
with
some
period
A
administered craving least phase. unchanged A reduces withdrawal substance.
In
cravings
treatment treatment
In
of
some
A
Compound
one a
the embodiments, suffering
an is
apathy, substance during the some is of
and
therapeutically
for inhalant, the In
substance optimal
administered
embodiments, which abstinence derivative. methylphenidate,
for
drug
selected
withdrawal some
or
from
the substance
of promotes
to
embodiments,
the the
irritability, In tapered from
the a
of
A
substance includes abuse
embodiments, at substance dose a patient
some relapse
abuse is sedative, in
relapse least Compound
or
selected
from In
from
to off to craving a susceptible may and remission. effective
the
some
one
is patient. by a embodiments, treat is recklessness,
after phase.
patient.
being
substance phase in dependence
escalated psychostimulant be
selected way the a substance. diethylpropion,
from
a
amphetamine, embodiments, tranquilizer,
the induced
conducted
the patient.
A remission
of amount amount
reduces abused
to
treatment condition a opioid In In In example In
drug at from
until
use
some some some some
poor by least in The
the
of of or of
in at is is is a a a a a
2014202047 11 Apr 2014 reduces trifluorobenzene, mushroom, Auditory the pleasure, nausea; physical medication. retardation, inflammatory xylene, Intelligence irritation; increased Lorcet, opium, anticholinergic some some some selected Compound (RCFT), embodiments, scale (HAM-D), dependence. Compound and chest dependence cocaine deficit
Compound craving
embodiments, from pain;
embodiments, embodiments,
hyperactivity
cannabinoid,
Percocet,
ethylbenzene, at vomiting; and and
from
and energy, Verbal nasal
depression,
Hamilton
A Neuropsychiatric
A
least heart agitation,
lysergic in
Scale-Revised
for mental
the the
reduces In In
increases
Lortab,
the the medication, A
crusting;
agent, cocaine. some
some Trail treatment one palpitations;
excitement Compound
pentafluorotoluene, Percodan, Learning patient cognitive
headache; the
strength;
acid
Anxiety
disorder the
cannabis, extreme the in
symptom Tramadol, embodiments, Making
irritability, embodiments, fluorobenzene, inhalant
at
an the
medication recurrent selected diethylamide
hallucinogen
In
least
(WAIS-R),
improves
patient an
Test antihistamine,
function and and Inventory,
decreased
some Scale A suspicion, IV
arrhythmia; vertigo;
Test
benzodiazapine over is one of
reduces sociability;
Tylox. heroin,
rating (RAVLT, from selected
at nosebleeds; (HAM-A), embodiments, sleep cocaine
(TMT, negative
a the is least
the rating
the
pentafluorobenzene,
Wechsler score
17
o-difluorobenzene, (LSD), attention sensation
dizziness; is and
scale and selected
drug
methadone, at
In drug disorders, counter from one
cardiomyopathy;
selected
Parts
a
withdrawal of least a less scale craving
Trials
some
subjective
of (ADHD-IV), cognitive
Beck
of
muscle the phencyclidine
carisprodol, nasal benzene,
of
Memory deficit hunger
abuse
of
A the from
one the
is
patient anxiety;
abuse medication,
embodiments,
pain; I-VII), and Depression
from for
amount
selected hydrocodone,
increased
stuffiness; drug
symptom
is
hyperactivity an relaxant, cocaine. and
B). symptom function selected
toluene,
bronchitis;
Scale-Revised
cocaine on is
psilocybin,
anesthetic,
1,3,5-triflurobenzene, of
and Rey Hamilton pychosis; heart tobacco,
fatigue;
cocaine, and at In
(PCP), from abuse
least
inventory
some
and
appetite, perfluorobenzene. facial
frequency of Complex In o-xylene, rating from attack; and a the of
the and
some
shortness a cocaine one
disorder; nonsteroidal is
cocaine
and nicotine,
an embodiments, marked alcohol, Depression the
drug
a an confusion;
pain;
fatigue, cocaine
Wechsler
scale.
oxycodone. (WMS-R), of
hallucinogenic
dilated antidepressant ketamine. embodiments, (BDI),
Compound analgesic,
pyschomotor
m-xylene, the of Figure
of
abuse dysphoria;
abuse
of feeling substance
euphoria;
Vicodin,
attention caffeine, abuse In
and
lack
breath; pupils;
apathy
Adult 1,2,4-
Scale some nasal
anti
Test
Rey
and and
the the
an of In p- In In of A is
2014202047 11 Apr 2014 role use beta therapeutically therapy. prolongs remission, reduced is in In one inhibitor include of one over Compound caused substance anticonvulsant, adrenergic sodium antagonist, (SNRI), serotonin characterized Compound likely activities efforts selected factor β -hydroxylase
the some physically having
by
symptom
obligations of adrenergic a
to patient
to the
longer (CRF)
treatment or important
because embodiments,
channel a
from
have a (NDRI),
to cut
norepinephrine
patient. use reuptake
period receptor a In and exacerbated
a A A
obtain
persistent down monoamine
by
selected of promotes period
been tolerance;
hazardous; some reduces
inhibitor, despite receptor antagonist, a effective sustained at
abuse at
of
blocker,
with glutamate of work,
a
social, the
In or least
substance antagonist,
caused
serotonin
remission inhibitor
then embodiments,
some control
from the substance,
of, at
having
and/or at
remission
by
one antagonist, school,
withdrawal; an
oxidase least
amount reuptake partial occupational
Compound dependence was
a least recurrent
or embodiments,
the recurrent an antagonist, adenosine of
calcium
substance
use;
exacerbated 5
one
in
recurrent persistent
(SSRI), intended; a early
-hydroxytryptamine effects or
one atypical
central the remission. use
inhibitor in
of and home; of
inhibitor
a
the
full
patient. the symptom
substance-related
the NK-1 the substance A contingency
least channel
on,
the receptor 18 use;
and of a
reduces
a
patient.
alpha
remission, physical
there
substance, recurrent
or methods substance
the gamma-aminobutyric antidepressant/antipsychotic,
or substance by
(MAOI), serotonin-norepinephrine the
one receptor a
recreational (NRI), recurrent
withdrawal
Compound In In great
the
adrenergic is blocker,
of substance.
use at antagonist,
some other
some In a
substance.
and/or
management
substance
least
substance
1
deal persistent
further early a
resulting some A and is a antagonist,
use
norepinephrine-dopamine
monoamine embodiments, often
social (5HT1A) agent
embodiments,
legal one
of a recover
is from A
partial activities
psychological agonist, embodiments,
time central
continued reduces
include symptom an In
taken use dependence in
In selected desire or problems; the
adenosine some
is and
acid a remission, a
some from antagonist, in interpersonal
corticotropin spent at failure a
oxidase and
in
are
situations in
central cognitive and/or least
the co-administering
(GABA)
reuptake
despite of its embodiments, larger the the
embodiments, from
peripheral given
in
a problem substance methods the
effects; to
and one patient
in
A2A Compound at
B sustained
tricyclic, or
unsuccessful
fulfill
remission
a
least amounts
a the inhibitor, knowledge
in behavioral
substance. up
peripheral agonist, continued dopamine
problems
releasing
selective reuptake inhibitor receptor which
at at patient
further
that
and/or one major abuse
alpha
least least
full
the the
an
of or
A is is it a a a
2014202047 11 Apr 2014 the receptor blocker valproate. receptor inhibitor, inhibitor, In embodiments, citalopram, embodiments, agonist, agonist, GABA agent selected embodiments, least antagonist and lamotrigine, embodiments, desipramine, selected embodiments, central embodiments, disulfiram. some atomoxetine. agonist agent sodium agent
some
levetiracetam. at
one is embodiments,
is
is
metabolism alpha
least
selected
the
channel
from a antagonist
from antagonist a
selected embodiments, other a an a
a
partial
topiramate.
central partial dopamine escitalopram, In
SNRI
In
carbamazepine, opioid mood doxepin,
one adrenergic
bupropion,
In the the the naltrexone
the the some the agent
some
from blocker
some
5HT1
at
at at other
selected or opioid at at istradefylline.
from at enzyme prazosin.
In receptor
stabilizer the least
least least
embodiments, is peripheral least
embodiments,
least baclofen, imipramine, least embodiments, some metabolism
In the
at
agonist, the agonist
agent selected olanzepine,
one
and and one agonist, one nimodopone,
least one some
one inhibitor,
dopamine
from
one at oxcarbazepine, antagonist, embodiments,
other
other naloxone. In fluoxetine.
other selected
other
is least
valproate, one beta other clonidine.
embodiments, a
some other
from
In duloxetine,
a
a nortriptyline, serotonin
agent
enzyme agent
other the
the agent partial adrenergic some risperidone, agent tricyclic the
a one
metabolism
agent
GABA
19 embodiments,
lamotrigine,
agent a
from at at
at
is
lamotrigine,
mood In agent other is
and is In embodiments,
is
least the least the In least
nicotinic
valproate,
a
5HT2 inhibitor,
is an some the
some
mirtazapine,
is
some SSRI selected synthesis carbamazepine topiramate.
partial
at
the
receptor
an
is anticonvulsant protiptyline,
agent
stabilizer, one other one
central and a
least
enzyme the
embodiments, atypical
at antagonist, embodiments, NRI
selected embodiments, carbamazepine,
agonist,
other
quetiapine.
other
the
dopamine least dopamine
agent is topiramate, one and
a
from
activator, antagonist
and
the selected an at
catechol-O-methyl-transferase
inhibitor In a
agent
one other and
agent
antidepressant/antipsychotic least is direct peripheral at from and carbamazepine.
opioid some and amitriptyline,
and the
an
selected least
other D2
venlafaxine.
β
trimipramine.
In is agent
one
the -hydroxylase
a
the an is opioid,
paroxetine,
NDRI propranolol. from tiagabine, the or
partial embodiments,
agonist carbidopa. lithium. the some one
receptor oxcarbazepine,
a
inhalant.
indirect at agent
other at at
calcium
alpha
is
from
adenosine
other least bupropion least
least
bupropion.
embodiments, a the dopamine
aripiprazole.
agent carboxylase is
amoxapine,
vigabatrin,
phenytoin, adrenergic
antagonist
sertraline, glutamate dopamine one one
In agent one
a In
In inhibitor In In In
In channel
GABA
the is some some some some some some some other other other
A2A
and and is the D2
In
at a
2014202047 11 Apr 2014 A reached therapeutically perfluorobenzene. triflurobenzene, o-xylene, In In inhalant, In In hydroxybutyrate, on embodiments, embodiments, embodiments, some from embodiments, some drug substance development acquisition Compound The substance Compound effective abstinence levodopa, after [066]
reduces some some some at some
methods
remission
of dopamine, embodiments, least embodiments, Also
in
abuse embodiments, embodiments,
amount a
embodiments, embodiments,
is dependence
m-xylene,
the
the from sedative, one A phase A.
carisprodol, provided selected
of
include administered
the the frequency
the the patient.
is is
effective
In
substance
the
bromocriptine, substance
of
reached
1,2,4-trifluorobenzene, opium, selected in at at at some at
In
extinction
Compound
the
least least
a the the least
least from
p-xylene,
some are administering tranquilizer,
is
In the the
psychostimulant
the embodiments, at patient. of the amount
selected
other psilopcybin, one in methods
in one some
other modafenil, from a use
least to
at Compound
relapse embodiments,
the drug amount a
at
other the phase
least
other
patient. by
agent A
patient. pergolide, ethylbenzene, embodiments,
least agent a other
of
patient In in of from
the
psychostimulant
of
a in other
agent
least
in the
agent some to abuse
hypnotic,
is of
treating
the patient.
other
agent
is hallucinogenic
the the A 20 the the acquisition,
In patient
acamprosate,
Compound
agent
until the agent is one patient.
is pentafluorotoluene,
the
patient. is embodiments, amantadine, Compound some partial
and
patient a
administered agent is
partial
an direct
methods at
other
a
is the In fluorobenzene, an the is
a is
desired
inhalant least
a embodiments,
medication.
determined some
opioid the
anxiolytic, In beta-phenylisopropylamine amount
In
a is serotonin
maintenance, agent, or agent nicotinic
A
therapeutically some one some
partial the mushroom,
indirect
A
further
is mazindole, gamma-butyrolactone, embodiments, therapeutic
agonist
selected
inhibits the
phase to
tapered selected
opioid of an
embodiments, embodiments,
the
5HT1 5HT2 Compound by and Compound
agonist
opioid, include In dopamine
o-difluorobenzene, pentafluorobenzene,
the
of
patient escalating buprenorphine.
some the
extinction, from an methadone.
tobacco,
off
substance from and response at agonist
antagonist
effective
illicit development
champix. a the co-administering least
after
embodiments, benzene, methylphenidate.
hallucinogen,
after
benzodiazepine,
A agonist A
the therapeutically the
substance. the
and promotes is gepirone. one
remission
and is
dependence
at
a
Compound
unchanged derivative. amount
amount ritanserin.
observed. period
least nicotine. In phase
In gamma toluene, selected In relapse.
of
1,3,5- some some some
one and
the the the
an of of In In of of
is a
2014202047 11 Apr 2014 reduces toluene, psilocybin, psychostimulant persistent withdrawal the rating treatment nonsteroidal hydrocodone, In often or home; HAM-A, Lorcet, opium, (PCP), embodiments, diethylpropion, amphetamine, deal substance substance. substance-related substance Compound frequency embodiments, antidepressant anesthetic, and 1,3,5-triflurobenzene,
recurrent
some WAIS-R,
of perfluorobenzene.
taken
scale.
recurrent
time
and cannabinoid, at Percocet, o-xylene,
BDI,
desire
dependence improves use of embodiments,
least an
A a
from In ketamine. is social
in
use hallucinogenic anti-inflammatory
In WMS-R,
reduces
and analgesic,
resulting medication. spent the dextroamphetamine,
some the
apathy
larger
one substance
and/or some
pemoline, of the
legal agent
oxycodone. m-xylene, Compound Percodan, or
the opioid
a in
symptom
embodiments,
cannabis,
at interpersonal
in
in
score amounts embodiments,
at
scale 1,2,4-trifluorobenzene, RAVLT, problems;
unsuccessful at
in
In
least the the an is least In
the
least
use a
some In
anticholinergic is mushroom,
patient
selected patient of mazindol,
from some failure p-xylene,
one and some one A beta-phenylisopropylamine in
of
benzodiazapine, In selected one the or
medication, Trials
situations reduces
embodiments, substance
and some
substance. Neuropsychiatric
of
Tylox.
over
problems
embodiments, patient and at substance
selected the to embodiments,
efforts
the activities
from
least continued
21 fulfill
I-VII,
ethylbenzene, Compound lysergic methamphetamine. embodiments, (-) a
cognitive from
in
longer
in on agent, one to
abuse In
from
the
ecstasy, cathione, caused
an major RCFT,
which by
cut In to
at
pentafluorotoluene,
Lortab,
some symptom
carisprodol,
the acid
patient
substance
over
the obtain
some
tolerance; least
down period an the in the
function
A
role
Inventory,
or
it inhalant
patient. and the antihistamine, diethylamide
fluorobenzene, phenmetrazine,
embodiments, drug reduces
is the the medication one
exacerbated embodiments, and the or
Tramadol, at
obligations physically
patient TMT, then of
hallucinogen
control derivative
least use counter
of substance,
rating of
withdrawal; abuse
tobacco, In fenfluramine. is
In
abuse
was the
at
despite and
Parts
some selected one some
selected
least
scale
substance of,
(LSD),
ADHD-IV,
is
by
intended; medication, hazardous; a a
pentafluorobenzene, heroin,
is
at
of
A the o-difluorobenzene,
muscle cognitive nicotine, dependence
embodiments, selected
use the is embodiments, alcohol, methylphenidate, the having
one work,
and is the is
the
from from
selected Compound phencyclidine the Compound
selected selected
effects
symptom
amount substance B. methadone, use;
relaxant,
substance, there
school,
persistent
In
HAM-D, recurrent recurrent
benzene, Vicodin, from caffeine, function In
and
a
of on,
some some
great from from from
is
and
the the the
an an of or or A A is a a
2014202047 11 Apr 2014 partial psychological the the receptor blocker, norepinephrine-dopamine norepinephrine management In other other one embodiments, embodiments, embodiments, continued activities and from and agonist, agonist, antagonist, enzyme GABA gamma-aminobutyric antidepressant/antipsychotic, adrenergic (MAOI), antagonist, (5HT1A) further agent
some
at at recover other fluoxetine.
duloxetine, agent agent
least least
selected remission,
include
synthesis a a
a embodiments,
a are antagonist, inhibitor,
agent serotonin partial
despite
antagonist,
monoamine
central agonist,
other one a is is from an
given
and mood problem
the the a the
the
reuptake adenosine co-administering other NRI is
from
In
its mirtazapine,
nicotinic
agent
remission dopamine Compound activator, cognitive
sustained a methods
and knowledge up
some a
5HT2 effects;
SSRI
a stabilizer,
selected
a
central acid agent
and/or
a that
peripheral a oxidase is
the catechol-O-methyl-transferase
corticotropin
inhibitor
selective
reuptake embodiments,
the dopamine
selected antagonist,
agonist, A2A (GABA) is
Compound at further is is
behavioral
β full a
reduced a or likely
NDRI
A -hydroxylase from
least and
the
characterized of partial B
a
tricyclic,
peripheral prolongs
receptor remission, a
inhibitor,
having direct (SNRI), alpha
adenosine inhibitor
venlafaxine.
from one and serotonin bupropion to
therapeutically include
bupropion. agonist,
β because an have releasing
-hydroxylase
dopamine
22
of
A an therapy.
the
paroxetine, adrenergic or opioid,
an
a a important
promotes
antagonist,
a
inhalant.
beta
been a inhibitor
(NDRI), period indirect treatment persistent
at by
and norepinephrine A2A sodium anticonvulsant, a
reuptake of
and
least
at GABA
adrenergic In
a caused factor In substance
sustained
D2
least
carboxylase In
receptor
atomoxetine. of
effective
some
receptor some social, one sertraline,
a remission dopamine disulfiram. channel
In inhibitor,
remission some agonist, with serotonin
and/or a one
metabolism inhibitor,
or inhibitor (CRF) some
other
monoamine embodiments, embodiments,
exacerbated
occupational use; partial receptor of
antagonist at
embodiments, reuptake
antagonist, amount
blocker,
a
recurrent
embodiments, early
least agent inhibitor, citalopram,
a in
agonist, glutamate and antagonist,
an 5 in
In In -hydroxytryptamine dopamine
the (SSRI),
the
remission. an
full enzyme some some one the adenosine antagonist, is
of
inhibitor a patient.
oxidase istradefylline. patient. by
opioid
a
calcium substance remission, physical and the a the
least a of a
SNRI
the embodiments, embodiments,
partial partial escitalopram, antagonist, central
a the
contingency recreational at an
at metabolism inhibitor, the
substance.
serotonin
one
least least
(NRI), In inhibitor methods
In
receptor In receptor selected atypical a channel
at
opioid and/or
use NK-1 5HT1
alpha some some some other early least
one one
1
In A is a a a
2014202047 11 Apr 2014 beta therapeutically perfluorobenzene. triflurobenzene, other o-xylene, In In In imipramine, olanzepine, clonidine. embodiments, central lamotrigine, least lamotrigine, some embodiments, embodiments, embodiments, some from embodiments, agent selected embodiments, least antagonist levetiracetam. carbamazepine, least agent agonist
some some some
adrenergic
one dopamine, embodiments, embodiments, one one
is is agent
selected
and
embodiments, from
embodiments, other embodiments, a a m-xylene, topiramate.
In
other other
risperidone,
and tricyclic carbamazepine, nortriptyline, mood
peripheral some is the the the
the the naltrexone
the
effective agent
receptor In
from bromocriptine, carbamazepine.
oxcarbazepine, an 1,2,4-trifluorobenzene, at at at agent
agent
at
at
In
some
at
embodiments, the
stabilizer least least least the
least
least
p-xylene, is some
atypical baclofen, selected
least
the
at
In the at the the amount and antagonist
is is
other
one one alpha embodiments,
and least one
protiptyline, other
some
least
at
embodiments,
dopamine
an oxcarbazepine, a one at
at quetiapine.
other other least
naloxone. other
selected
agent one calcium
antidepressant/antipsychotic
from
valproate, least pergolide,
ethylbenzene,
least anticonvulsant agent other adrenergic embodiments, of In
other the
valproate,
other
other agent agent
propranolol. some least
agent
is
one
amitriptyline, other at
metabolism agent is
23 the and
agent
from
least
the channel
the agent agent is one is embodiments,
pentafluorotoluene, the In In other and is amantadine,
partial
a the trimipramine.
and
agent is
receptor partial
an
at
some some direct
methods other one
topiramate,
topiramate.
fluorobenzene, carbamazepine
is is the
partial is
the inhalant least valproate. selected
agent
a
In
opioid the
the other blocker
enzyme
is serotonin sodium
embodiments, embodiments, or at agent nicotinic some
amoxapine,
partial the one
indirect dopamine least central further antagonist is
mazindole,
agent the
agonist selected
from an selected
opioid embodiments, other channel In
inhibitor
selected In In at tiagabine, one
5HT1
5HT2
selected
agonist opioid
some is include least some some dopamine
alpha o-difluorobenzene, pentafluorobenzene, and
phenytoin, buprenorphine. the other
D2
agent
from desipramine, methadone.
the the
prazosin.
from blocker and
one
agonist antagonist
embodiments, embodiments, central embodiments, agonist carbidopa. lithium.
receptor from
adrenergic champix. co-administering
at at agent
benzene, from methylphenidate. other is
vigabatrin,
benzodiazepine, agonist the
least least
the
selected
nimodopone, gepirone. or
aripiprazole. lamotrigine, is at
agent
bupropion,
antagonist peripheral ritanserin.
glutamate one one
In a In least
In doxepin, In In toluene, selected In
agonist GABA
the
the the 1,3,5-
is some some some some some some other other from
one and and the
In
at at at a
2014202047 11 Apr 2014 reduces reached recurrent use; unsuccessful related recurrent the A therapeutically maintenance, In In hydroxybutyrate, in Compound effective abstinence levodopa, symptom exacerbated social, activities dependence some social cocaine dependence embodiments, embodiments, a after [067]
patient. inhibits larger some some
Compound
remission
and
or
embodiments, occupational Also
legal
use
in
at
amounts embodiments,
amount interpersonal
embodiments, cocaine
to of the physical
the
the from
least A In
resulting
in by carisprodol, on,
provided obtain
cocaine
problems;
efforts
extinction,
development administered
cocaine the the
patient. some
the is effective A the
or
substance
one
of
reached
use opium,
or promotes Compound
Compound
patient
cocaine. withdrawal
or the Compound
or
to over embodiments, in abuse
symptom in
are
the
problems
use recreational psychological cut In the
a
and cocaine,
the
situations amount
and
failure
psilopcybin, methods a in
selected some
modafenil,
Compound use
down to longer of
Compound
and is development In the continued
amount relapse.
A A the the
continued some by
A
from
of patient.
dependence embodiments, to reduces reduces of
use caused
or
patient acquisition
in
the period
in of
from
cocaine fulfill Compound activities
the
the
control
embodiments,
of the which cocaine. treating
patient. The problem
hallucinogenic cocaine
A
24 A
at patient
acamprosate, tolerance; or Compound
the in
until major then cocaine, of
despite is
methods reduces least
exacerbated
the abuse
it cocaine the
administered frequency
selected
phase
are
at
was is
a
the In A.
use In patient
is
role
that least
desired extinction one physically
given
determined some
knowledge in some
or
intended; amount withdrawal;
In
the
despite in
include
at obligations
A
use;
phase is the one
recover from some
the
mushroom,
at
of Compound up
is least embodiments, gamma-butyrolactone, by therapeutic
likely embodiments,
patient phase
least a
patient. relapse to
tapered
or
of
phase
the having attention hazardous; great embodiments,
administering is
there
the
reduced
by Compound one
from
of
selected one
effects to the of at
escalating patient
selected
deal in
having in
work,
is
In tobacco, cocaine have persistent off
A symptom
cocaine symptom
response its
the the
a
deficit some
reduces
because
persistent
of the of
after recurrent
effects;
the
from
patient. patient.
after school,
been
time
the A
a to the therapeutically from
dependence
embodiments,
the Compound
is and is hyperactivity persistent
remission the
is of or
cocaine.
acquisition,
at of
a often
Compound
unchanged
is of
caused amount
important observed.
period
abuse recurrent recurrent or least nicotine. patient desire cocaine- gamma In
In spent cocaine cocaine
home;
taken some some
one
of,
or or or of of In in in A is a
2014202047 11 Apr 2014 withdrawal pain; beta therapeutically therapy. prolongs remission, the increases inhibitor include Inventory, increased some confusion; attack; shortness a disorder; GABA anticonvulsant, adrenergic sodium antagonist, (SNRI), serotonin characterized Compound and function least cocaine. factor β
-hydroxylase marked
amount TMT,
adrenergic
one
dysphoria;
embodiments,
dilated metabolism (CRF)
treatment
channel a
rating
a (NDRI), euphoria; In at feeling
appetite,
of of norepinephrine Parts
reuptake
period receptor and In and nasal
a A selected
least and
some
monoamine
breath; ADHD-IV, by
promotes
pupils; some
inhibitor,
a receptor scale antagonist, a effective sustained A
at
frequency of
one blocker, irritation; and cognitive
with glutamate of
a and embodiments, pyschomotor
least increased
enzyme physical antagonist,
from
serotonin
remission
inhibitor
chest is negative embodiments,
the nausea; craving B. at
remission
selected
one antagonist,
an
fatigue, oxidase least HAM-D, amount reuptake partial In
a Compound
pain; nasal
inhibitor, function of
an antagonist, adenosine of and some calcium energy,
vomiting; 5
subjective one in
for cocaine (SSRI), a early -hydroxytryptamine
retardation,
atypical
mental from the central the remission. lack heart crusting;
inhibitor in
of embodiments, cocaine. of
inhibitor
HAM-A,
a
the
rating full Compound
patient. the excitement
a
NK-1 contingency least of channel
WAIS-R,
A 25 GABA receptor
use palpitations;
a
strength;
a
patient. alpha pleasure, headache; remission,
symptom
methods gamma-aminobutyric
antidepressant/antipsychotic,
reduces
(MAOI), serotonin-norepinephrine recurrent
scale. one by receptor agitation,
(NRI), In
BDI, In In
adrenergic synthesis blocker,
the
antagonist, A some other WMS-R, and some the
some In decreased
depression,
management
of
In improves vertigo; 1
patient.
at apathy further early
a
some
A Compound a antagonist,
sociability; arrhythmia; nosebleeds;
norepinephrine-dopamine
cocaine
extreme
some monoamine embodiments,
(5HT1A) least embodiments, agent embodiments,
a activator,
partial
agonist, embodiments,
RAVLT,
central
scale
sensation dizziness; include
an embodiments,
In
a one irritability, selected abuse
suspicion,
score
adenosine
and A
acid some a remission,
less
antagonist,
from
nasal cardiomyopathy;
corticotropin a reduces a symptom oxidase and
partial
central cognitive
Trials and
the co-administering of hunger (GABA)
of reuptake
the
the anxiety;
embodiments,
from Neuropsychiatric
peripheral
stuffiness;
the
pain;
a methods the dependence. sleep and
A2A Compound Compound at
I-VII, B dopamine the sustained tricyclic,
or
patient
remission
and a
least of
a inhibitor, craving
bronchitis; behavioral
peripheral agonist, dopamine disorders, pychosis;
cognitive releasing
selective reuptake inhibitor
receptor
cocaine
fatigue;
RCFT,
further
one
facial alpha on heart
full the
D2
for
an
of In
A A at is a a a
2014202047 11 Apr 2014 the receptor blocker valproate. receptor inhibitor, inhibitor, In atomoxetine. embodiments, citalopram, embodiments, agonist, agonist, embodiments, agent selected embodiments, least antagonist and lamotrigine, embodiments, desipramine, selected embodiments, central embodiments, disulfiram. some agonist agent sodium agent
some
levetiracetam. at
one is embodiments,
is is
alpha
least
selected
the
channel
from a antagonist
from antagonist a selected
embodiments, other a an a
a
partial
topiramate.
central partial dopamine escitalopram, In
SNRI
In
carbamazepine, opioid mood doxepin,
one adrenergic
bupropion,
In the the the the naltrexone the
the the some agent
some
from blocker
some
5HT1
at at
at at other at
selected or from opioid at istradefylline.
at prazosin.
In receptor
stabilizer the least least
least least least
embodiments, is peripheral
baclofen, embodiments,
least imipramine, least some embodiments, metabolism
In the
at
agonist, the agonist
agent selected olanzepine,
one one nimodopone,
and and one agonist, one one
least some
one
dopamine
from
one at oxcarbazepine, antagonist, embodiments,
other other
other naloxone. In fluoxetine.
other other selected
is valproate, least
one beta other clonidine.
embodiments, a
some other
from
In duloxetine,
a
a nortriptyline, serotonin agent
agent
enzyme agent
other the
the agent agent partial adrenergic some risperidone, tricyclic the
one
metabolism
agent 26
embodiments,
lamotrigine,
agent a
from lamotrigine, at
at
at is
is
mood In agent other is
and
is is In
embodiments,
least the least a the In least
nicotinic
valproate,
a
5HT2 inhibitor,
is the an some direct
some
mirtazapine,
topiramate. is
some SSRI selected carbamazepine
partial
at
the
receptor
an
is anticonvulsant protiptyline, agent
stabilizer, one other central one
and a
least
enzyme the
embodiments, or atypical
at antagonist, embodiments, NRI
selected embodiments, carbamazepine,
agonist,
other
quetiapine.
other
the
indirect dopamine and least dopamine
agent is topiramate, one
a
from
antagonist
and
the
selected
an at
catechol-O-methyl-transferase
inhibitor a
In agent
carbamazepine. one other and
agent
antidepressant/antipsychotic least peripheral is direct at from and
opioid
and some amitriptyline,
dopamine
and the
an
selected least
other D2
venlafaxine.
β
trimipramine.
In is agent
one
the -hydroxylase
the an is opioid,
paroxetine, NDRI propranolol. from tiagabine, the
or
embodiments,
agonist carbidopa. lithium. the some one
receptor oxcarbazepine,
a
inhalant.
indirect at agent
other at
at
calcium alpha
is
from
agonist adenosine
other least bupropion least
least
bupropion. embodiments, a the
aripiprazole. agent carboxylase is
amoxapine,
vigabatrin,
phenytoin, adrenergic
antagonist
sertraline, glutamate dopamine one one
agent one
In a In
In
inhibitor In In In
selected In channel
GABA
the is some some some some some some some other other other
A2A
and and is the
In
at a
2014202047 11 Apr 2014 triflurobenzene, mushroom, pemoline, the psychostimulant psychostimulant patient. therapeutically perfluorobenzene. o-xylene, In In hypnotic, In In hydroxybutyrate, embodiments, embodiments, embodiments, some from some selected dextroamphetamine, and some dependence Compound cocaine Compound effective abstinence levodopa, agent
some some some some
a
dopamine, embodiments, embodiments,
beta-phenylisopropylamine embodiments,
is
medication.
In dependence
from embodiments, embodiments,
embodiments, amount
embodiments,
m-xylene, mazindol,
selected an from some A A lysergic carisprodol, on,
administered the
is the Lortab, the anxiolytic, effective
or
bromocriptine,
agent, agent cocaine unchanged of embodiments,
1,2,4-trifluorobenzene, opium, at at at
In withdrawal
Compound
the
least the and acid from (-) least
least is p-xylene,
the some
In
is Tramadol,
an
the the
the
reached at
cathione,
at
the amount methamphetamine. a use
other psilopcybin, some
diethylamide
one
opioid, hallucinogen
least other modafenil, beta-phenylisopropylamine
least to at Compound amount ecstasy, embodiments, and
after
at by
the least
other
agent from A the
one pergolide,
ethylbenzene,
embodiments, least agent other in
the
of
an patient and in heroin, a remission
Compound other
hallucinogen, the
of secondary
least the
agent derivative
patient. at
is
phenmetrazine, illicit
fenfluramine. other
agent is hallucinogenic
Compound
A 27 the least patient.
(LSD), patient acamprosate,
is
until the agent one methadone, is pentafluorotoluene,
the
is
amantadine, partial
selected
administered agent substance. is from partial one
an In
methods
other In
a
substance A fluorobenzene,
the is
is the phencyclidine desired
inhalant some
some
treats secondary In determined an
opioid the is
cocaine A
is serotonin
agent nicotinic drug
inhalant, some
In
from
is
partial the mushroom, hydrocodone, embodiments,
derivative. methylphenidate,
further
selected
embodiments, at mazindole, gamma-butyrolactone, some therapeutic tapered
is agonist selected
least to
of selected opioid
In dependence embodiments, psilocybin, selected
substance
the
5HT1
5HT2 by
embodiments, abuse agonist a include
(PCP), some o-difluorobenzene, one
pentafluorobenzene,
sedative,
off patient escalating buprenorphine.
from methadone.
In tobacco,
from
from
symptom and
response
agonist antagonist
from after
the and is
some
champix. embodiments, the
co-administering
and
in benzene,
is methylphenidate. a
selected after
psychostimulant
benzodiazepine,
a oxycodone. a remission the diethylpropion,
the therapeutically
reached hallucinogenic
ketamine. amphetamine, embodiments, tranquilizer, the drug
and gepirone.
the
is
of
patient. a
amount
amount ritanserin.
observed.
period
abuse
opioid nicotine. In of In gamma toluene, In from
in
abuse 1,3,5- some some some
from
and
the the
of,
In In In of of of In
is a a a
2014202047 11 Apr 2014 medication. trifluorobenzene, powders, parenteral the therapeutically mg/day. per Percocet, inflammatory xylene, other of or in its cannabinoid, anticholinergic some some combination limitation, and about about and dependence, acceptable corresponding esters, [070] [069] [068]
dependence certain example pharmokinetic
day. following other the
embodiments,
0.1 7.0 embodiments, appropriate
esters, A Pharmaceutically One
In ethylbenzene,
disclosure
Percodan,
sustained milligram to
therapeutically embodiments (e.g., Compound factors.
other
derivative
of
and
about the
cannabis, with hydrates, In
routes:
neutral
ordinary medication, without effective
intramuscular, age
without agent, embodiments
some
properties
pentafluorotoluene,
3500
composition In
release
and the at per of and
the
agent. oral, may
A certain
undue benzodiazapine, this solvates, fluorobenzene, least inhalant skill embodiments, kg Tylox.
amount mg/day, an a relative will limitation,
acceptable medication
therapeutically effective
formulations, (mg/kg) systemic vary
an
The application. are in antihistamine,
be one
embodiments experimentation
the the intravenous
over is
superior of and administered health Compound and widely while
amount selected
art
Compound pharmaceutically
body
take
amount (e.g., derivatives
prodrug
are is
the of
pentafluorobenzene, the 28
in
of o-difluorobenzene,
carisprodol,
selected treating depending with the
solutions, In weight
other the comprised
is effective
counter transdermal, from
drug or
A a a general, about
form
of forms.
subject,
respect
subcutaneous). as A may therapeutically and muscle
embodiments
benzene,
per pharmaceutical include such of for Compound
from
of
1.0 in be suspensions, medication,
amount
on
abuse treatment day The tobacco,
tablets,
by to acceptable of, reliance the
diseases to derivatized
the an intranasal
relaxant,
at
to about toluene, acids, way derivative potency in
least
is anesthetic, about severity
1,3,5-triflurobenzene,
for and
general, pills, Compositions A it
nicotine, alcohol,
or effective upon will of
10
is
a bases, one
elixirs, compositions and
or prevention
perfluorobenzene.
50 prior excipient. o-xylene, mg/kg/day. 70 of example about or
a
be capsules,
is
personal
of characteristic a mg/kg the
by nonsteroidal kg
an able selected
Compound
to
pharmaceutically an enol caffeine, Vicodin,
the
amount aerosols,
70 compound suppository)
human
formulation. antidepressant
analgesic,
to
to m-xylene,
body and addiction can,
of
ethers,
Acceptable semisolids, knowledge
ascertain Therefore, about
by
addiction such
without is
by is
Lorcet, opium, weight
or one
to 1,2,4-
A
from from used
anti
way
that 700 and any
the
an or of In p- or in
a
2014202047 11 Apr 2014 without milk, magnesium be, treatment pharmaceutical valuable moreover, nitrogen, mineral those Pa. starch, gaseous and excipients specifically 75% composition factors example, agents sweeteners, form. dextrose solutions, described [071] [073] [072]
:
for do Mack
and of Inert
w,
or cellulose, known The The Solid not
example,
limitation excipient
and petroleum, oil,
substances. carbon upon antioxidants.
the
in Publishing
or with include
are,
contain adversely gases required. pharmaceutical amount stearate, colorants,
will A.
pharmaceutical glycols. like. sesame
in
to
the by composition R.
dioxide,
the talc, comprise a those any
suitable
that
way
Liquid type water, Alfonso by
preservatives, single
of
animal,
sodium
Compressed affect
Company. Other remainder
solid, glucose, is flavorants,
oil, In way
Compound of of of
generally
nitrous
certain
ethanol, and
for
skill example formulation,
unit from
Remington etc.). the
preparations
of suitable liquid, vegetable is stearate, excipients
this semisolid
lactose, in administered therapeutic example
dosage 10% oxide, embodiments,
salts
being the
Preferred purpose gases available glycerol,
A solubilizers, and semisolid
pharmaceutical in w art glycerol
’
for
s
sucrose, or
include 29 size etc. without
to the may excipients
form Pharmaceutical of can the and
varying synthetic
are 90%
benefit
to pharmaceutical of
propylene composition
liquid in be
by
without or, excipient one monostearate, by
a ad
they by
a
w gelatin, used limitation, stabilizers, unit
way
single way in
of way the of libitum
may of
origin
carriers,
can the
dosage, the to skill carriers
the of
osmotic
of glycol limitation
of
be
malt, disperse
unit contain
case example compound,
may or Sciences
compound. example
example in
selected
when (e.g., non-toxic,
sciences.
wetting
the
sodium dosage kind excipients.
rice, particularly of and
and vary pressure,
art. still
the
an peanut
relief
water, and of
various their
flour,
and 1985, and
widely from
aerosol compound
preferably
other form chloride, excipients agents, In
Such
without aid
without
without
general,
buffers, formulations of oil, chalk, for 17th
saline,
Preferably
administration, therapeutically for oils,
for excipient depending
symptoms
composition, example soybean
emulsifiers,
ed. continuous dried
limitation,
in injectable 25% limitation silica limitation
including and
masking the
aqueous
Easton, aerosol
other
skim final w
may
gel, and oil,
the are for
to is
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expected, plasma urine
slow Cmax
93.8%.
and
was (57.9%) after
to
of
with Following In The than with
dosing doses. acetylator women.
and
be 1
the no
humans, were
approximately for
a the
pharmacokinetics was
2, a very in meal
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In significant In no
5.01%
administered
of 40
over
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men. pharmacokinetics
T1/2
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difference nepicastat
mg
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The
greatest was
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the
AUC (-100
slowly
48 1.4
difference population. 10
the
an 23% in
feces.
hours.
the hours radiolabel percentage
and -14
in with
the unidentified Tax,
in with
hours),
of
eliminated. AUC although subjects greater the
hours. women slow
slow
the nepicastat in of multiple
After for
The in
nepicastat, Tl/2. the
a
pharmacokinetics
was
of the
the acetylators, most acetylators was
40 radioactivity
terminal
in
No
of 30 fasted with
f0 was [ C
14 plasma
polar higher women radioactivity
mg
max
the recovered
C]-nepicastat, days, significant dosing There
likely
after approximately the
and
state. tablet total
an fraction Tl/2
the fast
in
concentrations.
than
a AUC
was than an due and
N-linked healthy
radioactivity single
mean
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taken
acetylator for
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differences
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no of rapidly in was T1/2
for
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(Ml).
total
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the
a nepicastat men.
total 40 significant
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radioactivity mg glucuronide
N-acetyl phenotype glucuronide recovery eliminated. There hours
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T 72 similar
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hours plasma than following the
was
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was
C metabolite
than
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in
max
present and with in
was no
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or plasma
in
single those significant to In 10
nepicastat nepicastat radiolabel
82.4% longer AUC men
average,
that
a a days in
to
were, meal, study
with with with
was
low and and and 3.5 for
for of in in in
2014202047 11 Apr 2014 mood, time the mg). testing block which were of Example 4 in informed inpatients Example Example doses less single subjects and daily and screening. generally [077] [081] [080] [079] [078] th
which
nepicastat day
duration 40 40
than
generally for
A
after from for
resolved 200
sleep, and In mg Five In mg) One Sixteen of
parallel
treated
4 2 4 3
participants well
that
treatment using
consent,
a a
mg both in 6
days, Eligible psychiatric
5 during
of of subject
study multiple-dose days
or
that to
demonstrated
tolerated following
dose well
the the
spontaneously.
cognition. with
a non-treatment-seeking group 800 or the
of
order.
double-blind,
study 6
daily
designed
volunteers well tolerated potential of
at
developed dosing mg
subjects 5 (n=12)
cardiovascular
nepicastat each of testing or
in
treatment over (dose
in
a Cocaine
participants
40 healthy
In
10 Phase order with dose
no
mg in to
receive studies treated
4 volunteers mg are
are
calculation
healthy differences atrial half-lives
placebo-controlled,
assess
200 of
level,
was
to
completed. men. is
dose admitted I with
nepicastat
maintain
administered
study
of mg
for ascending (n=4) significantly arrhythmias
and
participants men. 31 the thyroid of ascending
cocaine-dependent of 8
complete
of
based methimazole.
days
of nepicastat. and effects from receive subjective
nepicastat,
the
The demonstrated nepicastat,
uptake a or doses
on blind. placebo. physical
at
doses
study
greater and
longer receive of
only within-subjects hourly outpatient the
of of nepicastat
intermittent
which effects
hydrochloride Treatment of utilizes daily
In
123 cocaine
with doses than
examination,
intervals, cocaine nepicastat
I, Reduction no single-dose volunteers
doses
treatment
is 200 placebo, significant psychiatric
to
a
of 10
on (0
dose-escalation
at
0 mg design.
of return right
to 5
mg,
providing
mg, cognitive each
(0
5,
salt) and of
14
developed
EKG, with but
Phase
mg, are
40,
10 bundle
uptake 10 hours.
impairment
dose
40 and of to
After significantly
mg, and mg, placebo 80
studied
pregnancy
nepicastat I
mg sufficient
function, baseline.
level mg,
medical studies, 100
20
20
branch On after design giving
a
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rash
mg, mg,
160 mg the are
for
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2014202047 11 Apr 2014 pharmacokinetics parameters pressure participants day cocaine safety exceed Cardiovascular expected. [083] [082] 14-minute
4. 2. 7. 6. 5. 3. 1 of .
further.
normal AST, be between About Have Have Have one Have Meet Be Be In preset function limits study; treatment
administered during
order met between English-speaking
intervals
cocaine-positive are may
DSM-IV vital hematology a a and
with
limits.
within
12 baseline
self-reported conduction, Blood
indices
in to tests 85-150
complete all
alkaline
with days
of signs participate place the
18-55
is
procedures cocaine
(creatinine 2 on Previous
samples
following
TR
safe,
80 are are days EKG
mm as
the so
and
phosphatase) years
criteria
mg the follows:
carefully
and required
that
history and
Hg urine 3 volunteers of
are
that in rd
chemistry
study are and
studies
admission; no
day
of
the
systolic cocaine and exceptions: extending studied. involving
demonstrates
for prior
collected
age; clinically
resting 160
of study,
of
within monitored for BUN)
cocaine
using treatment have
who < to 32 mg laboratory and
each
is
3 Based
admission; participants
pulse the
one < x not the
for
shown nepicastat. significant 45-96 cocaine are
a) the 2 dependence; participant
inter-dose using x analysis
year. clinically be liver administration
not
with between on upper the
mm tests administered that
existing seeking
by
upper function
continuous 0
must:
limit
Hg mg 6 arrhythmias; the that of
The
normal to doses 50 interval
the
diastolic; limit nepicastat
IV
treatment complete information, are
of and
effects
tests
pharmacokinetics route
normal,
of of
within
of if 95 EKG sinus
to 32
cocaine, (total normal;
cardiovascular
bpm,
and this
of 1
mg
and
at the
rhythm,
and normal hour
and
nepicastat
the provide bilirubin,
criterion cocaine no
blood again
study.
frequent
b)
time and may interaction
kidney (+/- clinically
pressures
on
of
at
stopping of
enhance given
must
The ALT, indices
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10%) 10
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3
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.
mortality, the would psychosis, Be history history organic Have Have Have Have Have Have Have abstinence, sterilized, antiretroviral autoimmune, current consciousness, 8.
pregnant PI, Potential Have admitting significant
a evidence evidence neurological
HIV any any
apply: though make
family of of suicidal brain
a
as previous sterile, history
suicide any
bipolar and medical birth
or
determined study
renal, participants medication;
physician disease participants
history
psychiatric
of of nursing. contraindications
are chest ideation/plan
control
or
untreated clinically or attempts
illness currently compliance or
medically hepatic, post-menopausal)
history evidence
pain, in psychiatric or
pills, Other by first-degree and dementia
or
may disorder
are
or
within the significant or and or
major the
symptomatic,
as
intrauterine epileptic
unstable adverse suggestive
females active
excluded PI; be
difficult;
brief assessed principal
for
disorders,
taking the assessed
which depression
relatives
study 33 infectious
physical
or past
heart reaction must medical seizure;
be
from by antihypertensive
of device, investigator. would
participation, three
have disease
using
by
SCID; either such seizure
of
examination
as participation clinical to disease; illness
early
months a
require
condoms, assessed as: a
cocaine, be diagnosis
reliable
or disorder
unable cardiovascular
including: hypertension,
interview;
assessed ongoing in
medication;
by
including
form or the
demonstrating
to
of in or
SCID; spermicide).
conceive judgment
AIDS, the brain
of
neuroendocrine, treatment by
contraception
study morbidity as
SCID loss injury;
or determined
(i.e.,
are of of
if
no All
and/or
or the
any receiving
surgically
clinically
or which
females
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by
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2014202047 11 Apr 2014 there blood protocol. higher 9. cooperate cocaine signs arrhythmias Criteria Stopping Stopping [086] [085] 10. 13. 12. 11
.
must below. opinion Have Have 2. Meet Inability Positive completion sympathomimetics; and alcohol, 1
.
are
can pressure than Diastolic Participants Subject Systolic
at for are
provide
asthma discontinuation any
Criteria Criteria behavioral
the with occur
of Discontinuation
Cocaine not
opiates, urine those or to
other end
the
of if study comply
participation below administered BP
or negative
BP
vital
the in of PI for drug
of
illness,
currently
>180
must study or and/or
>120 expectation
manifestations procedures). Further study. administration the
signs
165 other screen with
criteria
pregnancy inclusion/exclusion mm participation;
continue
condition,
mm mm the
Following are
study
use is abused Participation (and
or
Hg admitting terminated
Hg Hg outside
alpha breath due of
sustained procedures; the
sustained
to
systolic
urine of drugs
or receiving to is
study or
meet
Initiation
use cocaine test of exaggerated 34
not
physician beta
tests
if
acceptable not of
indicating
physician for and any
inclusion
initiated
for
psychotropic agonists,
criteria
delivered before
cocaine. 5
toxicity
of 5
100 minutes
minutes would
the
response
mm ranges: study
because
illicit halts theophylline, if following criteria
as
(agitation,
preclude
there In or Hg
medications,
part or
entry,
continued
use
more;
resting addition, more; to
diastolic.
transient
in of
are of cocaine,
events upon
safe this order
cocaine,
psychosis, or
clinically pulse
protocol; cocaine other and/or
hospital repeated
which
occur: increases to These described
<130
remain cocaine,
successful
in
inability delivery) admission, significant values
the doses bpm
in
in
vital
and
the are
of to if
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careful 3. included
age and with
availability Medications
doses Participants Heart Doses Cocaine Cocaine Cocaine
(or criteria using
serotonin exacerbation
about to for
and severe
monitoring
Benzyoleconine
between to
by
benzoylecgonine
who because
rate Subject are
much
increases routes
sessions daily;
the
disease. is is produces were
90
of modest >
administered
take cardiovascular administered
use
into (220 are
medical min. higher
beta
this Selection typical of selected
of
of of required consisting
in presynaptic administration asthma
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agonist
group their compared prototypical relatively
participants age
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than The and
intervention daily
by primarily
x Criteria
underlying
at other IV, is to 0.85) those principal
medications)
medications
plasma
of
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so low
to
storage 10
high
to with minor
bpm
proposed stimulant
following that
35 availability
tone,
amounts
doses to 40
used doses, in
esterases
patterns risk
clinical
disease; avoid
sustained mg and active the produce
metabolites
granules.
evident
and cocaine
of
for
are case in
careful
here
death. effects 20
that enrolling
administration cocaine.
single
cocaine HIV are that is effects
participants excluded mg of
more
for
as
have
complete.
participants by
an on
screening
are
Cocaine administered increases by are 5 disease
doses
the These adverse
intensive minutes average
participants dependence. been
of inhibiting not excreted
IV
due cocaine
and
known
with associated
route potential has of are
Cocaine of
event. in
or 250 in
up to
interoceptive
cocaine, potential
heart
renally.
more. at a asymptomatic excluded
these
the
to to potential
with
mg to are short Participants 13
200 avoid
uptake be is rate
min toxicities to
with
psychomotor studies undiagnosed
metabolized
affected and elimination mg volunteers,
500
and
intervals. exposing to
seizures
adverse of effects. in
by
mg
blood avoid
have
with HIV self DA,
the are by or
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devices the study
of Treatment FDA.
participant.
No Ascending By Experimental Following 3 Cocaine To for from
treatment drug rd
volunteers
though
procedures,
Doses
with reduces
day starting pharmacokinetic documentation control involving were the available)
This use in illicit to using
of
of
study daily those
for
allow at
above
episodes pharmacokinetic
not
treatment approach DA, consent,
with doses at each
the nicotine IV and thus a
Cocaine
urine
a sessions cocaine studies.
that
us syringe are observed the lower use synthesis
160 DA-mediated dose
prescription
nepicastat. of to
far.
of based
records also
risks and in
mg
participants nepicastat reference interactions also
at
exposure,
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is
pump, Rash breath each conferred
asked
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conducted and assessment is heart
changes
NE,
the dose
continued incidence
drug
drug NIDA
alcohol which
reduce
is to side Because increasing (0
36 smoking
are are rate
combination obtained the the
wear a mg,
level
use use. greater
effects
expected
’ in required or
s at rewarding
administers and trials
the
level
of 80
IND
these
for for a cue approximately was in of
Some nepicastat
the
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from
risk
4
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exposure. for
for
prohibited
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to
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for experimental
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mg effects CHF,
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NIDA nepicastat the
rash,
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increases device paranoia does
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Participants
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mg) but
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of a
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is
cocaine hours this
not are this. by occur. may screening and and and the of
administered
of
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first and
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Because
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41
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46
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Indeed,
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mg.kg
of
modulation
least
This
and
sympathetic the
4
of of by
the
devoid cerebral
system. tissues
days and of
concentrations
stores
hemodynamic
dose-titration. synthesis
noradrenaline is,
magnitude which attenuate, carvedilol alterations
in contention
represents
1 decreases sympathetic
),
to
β of procedure.
perhaps, uptake SHRs, in -blockers, mortality. of
produced
those
without
of plasma
overall of dosing results cortex of
drive
this
and
but the the the
or of in in in in is a a
2014202047 11 Apr 2014 NMR receptors, uncorrected. precedence ketone. using primary thiocyanic Example hydroxylase over impact Point in KCN) hydrochloride, hydrogenated associated and system standard. spectrometer sequential azide 95% (LAH, chloride (available [0136] [0138] [0137]
THF.
natriuresis.
ee), β
Apparatus (+)- afforded
-blockers
spectra Nepicastat (-)-1 Melting The or to thereby
amide.
with
6
The The that
may chemical 1 with IR from acid.
5-(-)-aminonitrilc.
5, of /?,25- diformylation preparation
2/?-.V-mcthy
and Mass which
ethylene
spectra the
enantiomer was
absolute
and have
were
converted over-activation points
a obviating
Reduction the is
Competing .V-mcthylcphcdrinc,
or previously chemical mixture
is spectra
that
converted
ionization)
the AlCft-catalyzed salutary
a may were recorded
a were in
Mettler
of it potent, configuration
product
I CH2CI2
ephedrine
by
the
(91.6% be were nepicastat enhances
of recorded of (9:1) shifts
determined
hydrolysis
described effects
nitrile a
to need
of of aminonitrile or
FB on
obtained Strecker Formation
selective
the of
at are the Finnigan value
treated ee)
a
81HT -65 for
on
to
on as
azide dopamine R-(+)-mesylate, 2-ethylaminopyridine) Friedel-Crafts given sympathetic was
Bruker
was
5-(-)-2
of amine 47 renal of
on a °C) dose-titration. a
in
with
reaction chiral Nicolet
the based
the
a and cell
with
and available in MAT
under of
Uni-Melt the
function followed
-tetralol.
(93-96% nitrile ppm
ACF300, either
chiral
with
orally levels the
dihydronaphthalene auxiliary upon
treatment
anhydrous
nervous TSQ70 SPC the (formaldehyde
(δ)
reaction
heterocycle
afforded a by a
followed
center conditions such
Thomas Finnigan the which, active from ee) FT-IR Mettler
Another by the
AM300,
(for in system.
chiral
was as
to
of subsequent tetramethylsilane same
the
of
was
HC1
comparable
renal inhibitor
via spectrometer. LSIMS)
give Hoover
accomplished
by cardiovascular MAT FP90
3,5 advantage reduction Terashima described
was
bisulfite agonism based above AMX300
reaction -difluorophenylacetyl to
R-(+)-tetralol vasodilation, 7.
8230 processor
Capillary accomplished
spectrometer. give of
treatment
The described upon amounts
of by
dopamine complex (for of of with
reduction
UV
5-(-)-aminc
using
or tetralone
Terashima as azide
nepicastat dopamine
disorders
literature electron
Melting and diuresis internal EM390 sodium 4a spectra
of
route LAH
with
(92 was
and
the are by 'H
of β- 3
2014202047 11 Apr 2014 NMR with measurements rotations were mixture reflux min) mixture rate the which mL) min temperature of H, 2H); CH2CI2 evaporated difluorophenylacetic dropwise a dropwise (-)-l/?,2S\'V-mcthylcphcdrinc °C followed and complete, at g, [0139] [0140]
solution
such
2-ethylaminopyridine
4.42. 1.16 (1.0
product
sufficient at
was and 2% MS at recorded
(CDCh) for was
-65
a to (200
was such was
mol)
acetonitrile-98% dried rate
m/z by
at (45 then
were a ethylene 0.7
of °C. (7?)-(+)-2-Hydroxy-5,7-difluoro-l,2,3,4-tetrahydronaphthalene. 5,7-Difluoro-2-tetralone.
distilled
under further as
a of
heated
cooled
rapid
mL)
in min) a
5,7-difluoro-2-tetralone 182
rate
mm
to in a over
were on -65 δ cooled rate The order
white measured
maintain 2.55
CH
(M and
reduced
a maintaining
Hg). to
addition. °C
1
MgSCL. gas at
to reaction in
as Varian performed
h, + 2 acid
to
1.0 C1 ). reflux
(t, added in solid,
an vacuo
to to
during
was
2 maintain The Anal. (111 J
a M internal
20 (100 maintain
(1.0 -10
a dry =7.5
pressure. in
Cary
gentle lithium bubbled for (73.6 mixture mM dropwise Evaporation
distillate on
(81.3 The
g,
which a ice/acetone the °C Calcd g,
L).
on
Hz, 1
a
0.98
Perkin-Elmer
KH2PO4 3 an 0.58
temperature h
g, organic
Kugelrohr internal and a
reflux. a
then UV-Visible g,
aluminum 2H),
internal
The
was Regis time 0.40 through SOCI2
gentle for
mole)
(23.0 The was
0.454 to mol) 48 treated
allowed
Ci
a
under allowed 3.10
stirred mol;
a
bath, (pH
resulting redistilled mechanically layer temperature
After
0
Chiral
light g, and reflux. H in (100 temperature
mol)
collecting
the 8
of
(t, 4.7) hydride F 126
anhydrous 70%):
with
reduced
Model and spectrometer,
2
after
was
yellow-green to O: -65 the
J=
suspension
reaction mL) to
in AGP
mmol)
at cool
the C, oily
warm
The
addition °C H anhydrous at 7.5 separated, stirring mp
1
2 was
(416
65.93;
below mL/min
O
material acid 100-105 pressure 141
to using
column stirred <-60
reaction acid Hz,
46
Et
mixture
in room (500 to
added
2 mL,
chloride °C;
polarimeter. O
was
for 2H), 0 Et was
H, °C. Leeman -60 chloride a suspension
°C suspension
(100
2
at dry Et washed
mL) temperature. (4.6 O °C boiling IR gave
0.416
4.42. 15
°C. mixture
20 in complete, 2 cooled After over 3.58 at
O
(125
(KBr) (0.3 ice-acetone h,
mL)
one x
solution °C. a
initially (1.1
a
After
the
Labs
mol) was Found:
rapid
100 (s, 2 dark the between mm with mL)
portion
h
was appeared.
to
volatiles was Chiral L) 1705 A 2H), of
with
addition mm)
dissolved
Inc. the in
the
oily
Hg)
an
rate solution AICI3
was was brine was A
added dropwise,
C, Et heated
bath
6.70
cm addition reaction solution
stirring,
internal Optical residue
2 90-110 to
to eluting
HPLC for 65.54; O added added added
were 1 (100 (154
give
; was 3,5- The at and
(m, (45
'H 10 of in
at a
2014202047 11 Apr 2014 NaHCO was warm reaction was with the After treated mp <35 The -20 addition Calcd CnHi solid dissolve 530 solution C, CHC1 The 6.60 δ deposited enantiomer separated Sodium [0142] [0141] 1H), 'H
1.70
65.47;
NMR
solids. °C. 85
mmol)
spectra ethereal complete,
°C. cooled z-Pr (m,
3.20
(87.1 3 2
5
Et for ); F (br
°C;
under
3 2
h was 2 azide A the 92.4% 2 of
O , of 2H).
O δ
O H, After CnHi
crystals and brine s,
the
3
(dd,
(,S)-(-)-2-Amino-5,7-difluoro-l,2,3,4-tetrahydronaphthalene (/?)-(+)-2-Mcthancsulfonyloxy-5,7-difluoro-l [a]
solution 2.13-2.28
S: R-(+)-5,7-difluoro-2-tetralol in
g,
The
(20 gave MeOH 5b for
solids. 5.39. in 1H), solution stirred
argon
25
anhydrous
332 reaction C,
(40.0 the
Anal. the
D
are the 2 ee J an stirring mL) F organic and
an +38.1°
2 1.76-1.88 50.37;
=
by were
O ethereal
mmol), mixture
ice
6S)-cnantiomcr
identical:
of (100 with A for
analytical 3
17.2, g,
followed dried Calcd S: (m, was chiral
small 3N
bath was 0.62 a
at
C, collected
phase
(c Et stirring H, mL)
2H), further
evaporated
HC1
an 4.7
50.37;
used over
2 = was
layer
for (m, HPLC. O complete amount mol) 4.61.
and
1.83, increased
mp
2.78-2.96 by maintaining sample: (1.78 Hz,
was
C (2
2H),
stirred
directly MgSCfi. with
10
io was
maintained H, the and
79.9-80.9
L)
was H 4b
CHC1
Anal. 1H),
Found: separated
min
L)
i 4.61. of
1.99-2.06 0 was
under addition MsCl are F washed
dried
(as
mp
added at was 2 EtOAc 49
rate
0:
(59.0 at (m,
3 5.20 in
Calcd ); identical: then
-65 Found:
determined
Evaporation
78.8-80.5
-65
the
C,
the
reduced 93.4% C, cooled in (37.2 to
2H), °C;
at to
and
°C
(m, g,
of with 65.21;
vacuo was added °C (m,
internal 50.41; achieve
next
for
0°
DMSO
320 [a]
hexane
to
3.07 C, mL,
washed and
1H),
ee 2H), (-15
CioHi added mp
brine 25
C pressure -68
50.41;
°C; step.
H, mmol) o to
at by
by
H, allowed (s,
55.3
for
of 84-85 total temperature -16.6°
°C) ,2,3,4-tctrahydronaphthalcnc. 2.63-3.08 a 6.67
give °C 5.47.
[a] (1 chiral (200
0
3H),
TLC)
(200 to rate 4.64. F the sequentially
Trituration 1
H, 2 25
L) g, for
using
0:
help and
dissolution, and o the h
(m,
°C;
mL).
480 to 4.65.
solvent
3.09 (c Found: to with +16.8° HPLC: mL) C, 3 whereupon The and
limit alcohol Et the
complete warm h =2.23, 2H). [a] an (m, 65.21; mmol)
3
(dd,
and
N
below
stirring and water 25 The spectra residue ice-MeOH
3H),
o the (c of gave C, (74.2 with 'H
to J=
quenched
-37.8°
Anal.
= H, dried the
CHC1 (10.9 seeded
a 65.38: over NMR
temperature hydrochloride.
approximately
was dissolution 4.15 1.86,
-60
small the
17.1 5.47. an until IN mL, dissolved for layers
3 Calcd (c (MgSOj. 5-10 g,
off-white °C. ).
added resulting HC1, bath (m, (CDC1 H, CHC1
solution the Hz,
= Found: 53.9 sample 47 a by
Anal.
5.42. were
clear 1.24,
min.
1H),
The 4.7, (S)- %): and the aq. for
3 of to to in g, A 3 ); )
2014202047 11 Apr 2014 pentane/CH volatile mL) mixture yield nitrile reaction the bath to mp the the resulting the 2103 hydrogenated give sequentially diluted solution carbonate showed extracted g. amine 54.68; 6.93 (m, filtration for enantiomer [0143]
250 obtain
C10H12CIF2N: remaining fine >280 mixture bottle
1H),
and and (d, dihydronaphthalene
(44.8
cm
hydrochloride mmol) product H,
J= oil with precipitate
was
heated ca. mixture
mixture treated 2.63 ’ a took drying
with °C; 1 in
(5)-(-)-(5, 5.51; ;
was
solution.
which
2 g,
8b 9.4
m/z C12 washing
10-15% had air. obtained.
[a] over in
H2O
(m, solvent 202 about
EtOAc are
Hz,
evacuated
(39.4 at
N, (9:1) over water 25 with
was been
was 171
C, Recycling
D
1H), was
mmol; identical: 50° 10%
filtered 6.37.
(1.8 7-Difluoro-l, -60.2°
1H),
54.68; Further
4
8a of
MgSO the formaldehyde afforded stirred removed
filtered
to
(M g) (150 h). stirred rapidly
2.83-2.92 C
The
Pd/C
(50.27
L)
starting
give
7.00 +
combined
and (8.50 87.5%): ). for
Found:
(c and
The off H,
mL), mesylate
4 and this water mp
for
.
=2.68, through (6 an
(dt, The 16
5.51; for
the starting and
chromatographed Evaporation in g,
recharged
moist
g, g)
>280 a
oil amine
amine 2,3,
extracted followed h (m.
vacuo J=
229
azide further 30 C, mp
(300 in
washed
51.2
azide
under N, pentane (51.3 sodium MeOH);
4-tetrahydronaphth-2-yl)(cyanomethyl)amine.
a (138
2H),
54.31;
min, Celite, 9.4,
solid
mmol) °C;
73.1-76.5
6.37.
2.5 50
free
gave mL) remained. to (101
mmol)
6a g)
1
2.5 with a
[a] by g, 3.14
L give
with
KCN
was with of
h bisulfite
extracts
Found: N2 which was amine
H,
Parr an stirred
0.53 was 25 g, was 'H a at Hz,
the
o
few
a (dd,
483
hydrogen
additional 5.52; atmosphere. transferred
EtOAc, as
80 NMR
°C; +58.5° on
added
white
pentane bottle (15.0 mol)
treated solvent 1H).
dissolved
solidified.
a °C, with
(7.12 additional C, J= mmol) Chromatography silica
with complex
[a] volatile N,
54.64;
(i/
was
and solid
25 cooled (60 g, Anal.
(c 16.7, and
ethereal
6
6.44.
D
with
H -DMSO) under
using to g),
5.35 230 (4
=
as -58.0° the psi) 2 to
added
O
then
(90.4
in
1.63, remove
H,
(30.8 5.0 Calcd
a The oil. x which pellets a TLC
a
to (2
mixture
mmol)
g colorless
for The EtOAc round-bottom pentane reduced
5.51; solution
250
HCI
Hz, of room anhydrous x
(c in
g, MeOH).
reaction
δ 6
g, Further
(5%
100
for
product.
spectra
= one 412 h.
of 1.79 quickly on
mL) 1H), (IN, evolved N, 230
was
placed 1.63,
temperature,
C
NaOH
After (1200 mL),
MeOH-CH
as pressure oil: 6.40. portion of silica mmol; io
(
mmol).
3.46 500 H
m, the
followed mixture
added. NaOH elution
Anal. for ether. CHC1 i IR
2
brine
each Combined forms in C1F
flask,
N2.
sufficient 1H), mL) mL),
eluent gave
(m, (CHCI3)
the 77.9%): a
and
gave
2
3 Calcd 50
N:
After
hour, (10.0
); 2 (The
(100
with
The 2.33 1H),
was Cl (R)- The The
and and and and
the the the
°C by 'H C, to
2 a )
2014202047 11 Apr 2014 NMR thioxo-l//-imidazole. treated was mmol) were reduced heated 244.7 2.14 262.7 Calcd 64.85; dihydro-2-thioxo-l//-imidazole. 3.80; C, 309 elution C14H11F2N3S: 9.3, °C; slurry and formamide and enantiomer [0144] [0145] 1H), 1H), 1H), 10%
53.32;
(M [V] then
the
2.4 evaporated,
2.75 7.05 (m,
N, acetone/CH2C12 3.70 cooled
(CDC1 for in
°C; °C;
H, was + at with
25 of
).
residue pressure. 14.42. Hz,
10% d
placed 1H),
C12H12F2N2: H, (m, reflux (dt.
5.44; the [V] [V] (5)-(-)-1-(5, (5)-1-(5,
(s, Anal.
-69.1° and
added
9b
KSCN 3 in
3.96:
) 1H), 25 25 J= MeOH/CH 1H), 2H),
column
2.15-2.28
C,
δ
D D ice/MeOH are
ethyl Found:
N,
in was (120 the Calcd 1.50
+74.9° -90.5° 57.72;
9.5,
(c
over N,
7-Difluoro-l, 3.03-3.35 8.29
Toluene identical: an 6.62 12.60.
(78.1
=1.18, residue
formate
dried
(br gave 13.24. 7-Difluoro-l, °C 2.4 with C,
ice
The for
C, 20
(c (s, H,
(m, (c 2 (m, 64.85;
s,
bath) C12
g,
Hz, 57.85; bath
(-15 Found:
Ci
min.
=
= the 3.80; under
1H), DMSO); nitrile 1:1 1H),
was
804 dissolved (m, 1H), 4 2H); 0.398); The 2.14, on mp
(48.7 Hi3F
1H),
product
°C) to under McOH/CFFC
2,3,
H,
to
After 3H), added 1.70 H, N,
mmol). The
13.3
64.4-73.6 spectra 2.74-3.05
give high C,
2 MS (44.7 DMSO). 5.44; a 2,3,
N3OS«0.25H
mL, 8.73 and 4-tetrahydronaphth-2-yl)-5-aminomethyl-2, 3.85;
14.42. silica
IR
65.07; (m, 'H above 5.19
N2
(br the
4-tetrahydronaphth-2-yl)-5-cyano-2,3-dihydro-2- in
51 m/z a
and vacuum
(18.05
stirred
44.7 (KBr)
(s, N,
NMR
g, precipitate. N,
for
for 1H), IN
The
reaction
s, (1
(m,
Found:
222 1H), 201 12.60.
evaporated °C; nitrile (m,
’
H, 14.45. Anal. the b kg) g, 1H); HC1 19
mixture 2.05 while
1H), g,
1593,
gave (i/
5.47;
604 2 4H), mmol)
[α] (M to 9.70
h, O: (5)-enantiomer column 6
62.1
-DMSO) Found:
(990 MS
(5.00 Calcd had
25 C, + give
and 6.94 (m,
).
mmol) C,
the
π /-BuOK
1630
N, (br 5.64
The
57.82;
mmol; +52.3° was
been m/z 53.57; to
Anal.
mL) 1H),
in the
primary (d,
an g,
packed s, 12.44.
for
C,
remove
filtered (m,
butyl
cm
J= 1H), in 17.2 stirred 291 δ
solvent
oil
65.14; 2.55-3.04 stirred
H, and C14H11F2N3S:
(IM ’ 2.18 Calcd H, 30.8%): anhydrous 1
(c
1H),
; 9.3
(53.2
The 3.92;
mmol)
are in (M 13.7 formate 'H amide 4.33; ethanol =2.12,
solid in last
for
Hz, (br hexane.
H, +
for then 6.90 NMR identical:
).
spectra
for THF, (br N,
g).
5.54;
135 N, (m, m, traces
m.p.
1H),
18
in
Anal. was
11a:
CHCI3). C12H12F2N2: 14.37. removed s, (d, THF (497 (240
13.39.
THF 1H), (t/ h, The 4H), min
302
1H);
N, Elution C, 7.03 240.7-249.2
J=
for 6 loaded
the
-DMSO) of
mp
mp
3-
Calcd
(935
mL), 57.72; mL) 12.53.
resulting mL, 2.47 at
3.22 (75
(Further
the
solvent, MS
9.2
solvent Found: (dt,
261.9 243.1 85 under Anal.
with
mL) mL)
as
was
(R)-
302 and Hz, m/z (m, J= (m,
for °C H, C,
δ a
2014202047 11 Apr 2014 then was tetrahydronaphth-2-yl)-5-aminomethyl-2,3-dihydro- by treated with prepared 4.96; hydrochloride extracts and solution solution 0.8 C -10.8° Calcd (m, dried (3.12 dihydro-2-thioxo-lH-imidazole Ci4Hi 58%): (5.2 sodium solvent stirred for [0147] [0146] l,2,3,4-tetrahydronaphth-2-yl)-5-aminomethyl-2,3-dihydro- 12.39. i 4
chiral H
nM)
1.5 3H), the
g) stirred
the
EtOAc i
6 5 g,
under
N, for F2N3S«0.25H C1F for
Nepicastat on h.
with
was
aqueous mp
(c potassium were solution
(30 of and
10.6 4.77 HPLC); by
2 Ci silica (5)-1-(5, 15 The
N 12.18.
LAH
under
= mL) partially several 4 EtOAc the 3 human
nitrogen
min Hi
S«0.35H 170 mmol) (m, washed,
salt reaction
1.43, 6 eluting
phase
addition was ClF
was in
1 and was 1H), argon H (3.87 7-Difluoro-l,
tartrate
°C; 2
THF (IC50 times
The O: (150
2
NMR
N which
stirred removed 2 added,
treated
O: and demonstrated DMSO), dried
extracted 6.84 3 C,
was with
S*0.5H g):
in
[V] (1
of mL) = spectra
56.07; without
C, (T
until then
an
9.0 mp
(m, M,
again had 25 for ethereal 5% (MgSO followed
with
D = 49.73;
and ice 2
under
± hydrochloride 34.3 2,3, O: 245 30
2H), 320
with MeOH/CH been
H, under the -11.0°
91.6%
0.8
cooled water
for bath evaporating min
ether 4-tetrahydronaphth-2-yl)-5-aminomethyl-2,
5.21; 4 C,
°C °K, to
mL, ),
7.05 mixture HCI nM) H,
reduced
10%
by dissolved
be the 52 49.33; and at
(dec); until
high
DMSO) (100-150
4.98;
(150
N, to
34.3 (c 0 10% ee
(s, a DBH. (IM,
McOH/CTLCfl
2 evaporated. °C
competitive (R) 0
C12 14.01.
a
1H),
vacuum
=
became °C
by H, pressure
[a] (nepicastat). mL), N, mmol) MeOH/CH to and -enantiomer
homogeneous
20 gave
in
25 δ
and
The 5.03; dryness. 1.59,
mL). 12.42. 8.57
D 2.07
chiral allowed
MeOH mL,
are Found: filtered +9.65°
the treated was 5-cnantiomcr
at freely
(br N,
and identical; (m,
inhibitor The
20
DMSO). Chromatography free
2
78 Found:
(2 C12 HPLC.
added
s,
12.33. (250 (IC The
C, to mmol) 1H), displaced (c
off,
The
(R)-l-(5,7-Difluoro-l, organic with 3H), x
amine
stirrable. °C solution come 56.11;
(200 50 =
125
resulting
s
mL)
washed
2.68-3.08
1.70, hydrochloride of
mp dropwise for = C, a
12.4
Found:
to
mL). mL)
saturated bovine to 25.1
(R)-l-(5,7-Difluoro-
(2.92
Anal. layer H,
49.80; 261-263
Anal. by
20 by the
DMSO); room was (br
5.10;
and Further with ± warming.
precipitate
co-evaporation
h The
free g, of was
s,
(m,
0.6 over
obtained. (IC50 C,
to temperature H, Calcd
the 9.89
the 1H).
Calcd N, solution
3- ether, °C; combined 49.44;
4H),
separated nM; amine
give
4.93;
(93% salt
mixture 10 14.14.
= residue tartrate
mmol;
2,3,4- [a] Anal.
8.5
min,
4.09 18.3 was was
for The
and
the for 25
N, H, 2a
ee of A ± o
2014202047 11 Apr 2014 mixture was purified was measuring ± produced parameter membrane parameter normal inhibitors Figure 0.5 CUSO4, administration and (DA)/norepinephrine approximately and enzymes selected established (without detection standard). stop concentration (final determined [0149] [0148]
0.6
- highly performed obtained cerebral
solution 1
volume nM)
Oral 3 The
milliunits
containing 0.1 from dogs enzymes
and shows inhibitor),
at sustained of
concentration-response logistic
the
preparations. The selective
and
by assays. activity mg/mL administration 280 the
neurotransmitter of cortex.
the from 2-3 conversion
of containing produced
samples
at of
SKF bovine radioligand the a nM.
200 and culture equation. pH table fold of
increases
nepicastat catalase, Sigma inhibitor inhibitor
catalase,
corrected of (NE)
enzyme
102698 The 5.2
receptors. more pL).
The Chronic
and were nepicastat describing
Binding medium
potent of
and
25 ratios
Chemical of
affinity remaining human
potent
K, tyramine
binding
at tyramine in of analyzed 0.1 Samples
(IC50S were nepicastat mM
receptors. 32
using
values produced 37
the
oral
DBH in
mM curve
Nepicastat and °C data
°C
of
the than at enzymes, peripheral plasma
added EDTA of
=
administration
in Co
the
for internal tyramine
assays in to
were eleven percent
for were
and were 53 interaction 67.0 dose-dependent to
nepicastat
the
0.125
vitro. to octopamine.
30 (St neuroblastoma
obtain These octopamine
dose-dependent
to
DA/NE
ascorbate
spontaneously R-enantiomer calculated and
- ± analyzed
incubated respectively.
Louis, showed
using the
40
arteries different activity
standards M 4.2 Moreover, and
IC50
min.
data 240
reaction NaOAc,
of
nM;
for
ratio. 4 MO). of
standard
values. nepicastat weak was
mM
(renal
suggest by
using
by μΜ Bovine The
was
thirteen with enzymes nepicastat
85.0
increases
cell
iterative and
reverse
suggesting
In the
hypertensive added mixture ascorbate.
DBH reaction 10
3-hydroxytyramine calculated
and
affinity Human
or the
conscious or
line ±3.7 that S-enantiomer
mM DBH fitted filtration mesenteric),
Cheng-Prusoff without at
selected
activity
long-lasting phase
to was
SK-N-SH.
curve nepicastat
to
DBH
in fumarate, and
was nM) for secretory
from initiate
to stereoselectivity.
normal In based
determined
tissue
SHRs, HPLC
a rats then quenched
a
techniques fitting the was
a
and are range receptors adrenal
non-linear typical
left
nepicastat
the
(SHRs)
on
appropriate
a less assayed is
0.5 a
DBH The
dogs
acute
using dopamine
(>
equation.
to substrate ventricle range (internal a of reaction controls
-2 glands potent potent by a assay,
using assay
4
other
four was also oral was
UV μΜ and and
the
by
h) of 4- is
2014202047 11 Apr 2014 mesenteric mg/kg, ventricle) with release. norepinephrine neurohormonal progression neurohormonal renin-angiotensin be heart DA increasing however, II, h CHF congestive cardiovascular sympathetic antihypertensive (vehicle) study, spontaneously such enzyme evaluation. angiotensin-converting adrenoceptor β [0150] [0151]
-adrenoceptor apart,
have
expected
levels
halothane, is as failure.
the Nepicastat Congestive
po, characterized been beginning responsible low
Previous
or have were
heart the artery. that
b.i.d. animals the
nerve
of potency to An shown antagonists,
hypertensive a
rapidly tissue (NE) tissues, systems appropriate
can antagonists decapitated,
reduce CHF. systems failure.
limited
effects
for DBH
alternative
On in heart was stimulation.
system
for
produce were to
biosynthesis by the 10
and DA/NE the
harvested,
reduce
enzyme
inhibitors,
conversion
used are tissue nepicastat indirect failure marked and days) Therapeutic
morning.
basis has
specificity, such weighed
dose
likely
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also and
renal
strategy
in
morbidity
been
ratio.
which levels of (CHF) a as (ACE)
Serum
ability activation
of
(SHRs) weighed,
the
in
attenuation
its promising
to via such
reduced At
of vasodilation, nepicastat. has
and
vivo ability favorably
increasingly
tissues
elevated that interventions NE 6 of inhibition
is This
to 54
inhibitors, to T as The
h
been a
and 3 randomly
NE
and
after attenuate biochemical
fusaric to leading directly
and
have placed of
stimulation to
approach
(cerebral mortality dopamine
approach
the of
simultaneous
and
potently normal
clinical the T alter Each the
precluded 4 of the
natriuresis
sympathetic
acid
in which cause levels
implicated
elevate
dopamine/norepinephrine third modulate
dopamine
the
assigned the
which iced pressor rat
has beagle
and
cortex, in modulate
SNS.
(DA). evaluated of that studies
natural of
was
dose,
were block 0.4
CHF
SKF-102698, potential α mortality tissue their
block
-adrenoceptors
and
responses
dosed M is
activation
the nervous to dogs. Inhibition
β mesenteric
unaffected the
in
Inhibition patients.
-hydroxylase formation
perchloric history
to currently
clinical the
diminished receive
levels the rats SNS
for
the orally
study
sympathetic in advantages
On
system were
perpetuation the the
to of
effects
is
of have
ACE
development of
of
the of
three by either
artery pre-ganglionic the of under CHF. acid, United
the
inhibition
treatment and
DA
DBH anesthetized
ratio angiotensin (DBH), aldosterone a
these
day drawbacks (SNS)
inhibitors,
effects
SNS dose
times,
frozen of
elevated and placebo over drive thereby Indeed,
clinical
States.
of would
in
these
with
(6.2 two
and and
left the the the
12 of of β- to in in in
2014202047 11 Apr 2014 perchloric the placebo were potent nepicastat mesenteric mg/kg the mg/kg ratios increases Marshall and (cerebral appropriate study, liquid and concentrations samples and fold, dogs) dogs. assayed [0154] [0153] [0152]
mesenteric DA first glutathione, SKF-102698
whereas
investigated
nitrogen
in in in in in dogs At The Oral
concentrations
by (empty
the the dose the SHRs, were Farms in cortex, SHRs,
the produces
acid,
artery,
chronic dose
DA/NE
HPLC were administration artery latter artery,
nepicastat, highest
on artery, of and
drawn, frozen
centrifuged SKF-102698 USA
produced capsule) of
(1)
renal
DA
left day randomly in
species.
stored
(mesenteric
effects using nepicastat. the
ratio
left in
normal
dose left ventricle Inc,
and
were 5, in SHRs. artery,
expected
6 ventricle
at
were at
ventricle liquid the or
electrochemical only NE. North
tested
of h
the of
Furthermore, at -70°C
assayed
assigned dogs. nepicastat
(1) nepicastat
dogs
after -4
nepicastat
left
14, and same
or 2.6, Each nitrogen The
increased biochemical °C
(100 Rose,
and
renal), until
ventricle) 11 and
cerebral were
3.5 and Animals the by
samples dose,
dog
and mg/kg to cerebral
55
cerebral
NY) HPLC analysis.
and
(14.5 ( stored
first
and receive
produced nepicastat euthanized left
was 3.2
2 the increased detection.
cortex,
1.1
mg/kg,
were
in harvested, were were stored
ventricle effects fold day
DA/NE using
dosed dose, at
cortex, SHRs cortex, fold
-70 Tissue either
treatment)
(in used
collected randomized
respectively.
dose-dependent
is
at electrochemical
b.i.d) increases the in twice
°C with
for
SHRs)
and more ratio
-70
respectively. Male and respectively. both
in
placebo NE until weighed,
ratio
the
5 the
pentobarbital
°C for a
by cerebral
potent
mg/kg and
in SHRs on
and day beagle
analysis.
by
until study. in measurement 5.5-fold,
14.5
tubes
to
the DA
(empty the
These 8.3, 95, for
placed
receive,
than
in and
analysis.
plasma increases
cortex detection. concentrations days.
DA/NE 4.5 dogs
151 Compound When 7.5 containing dogs) On
dogs
data 3.5-fold
its
capsule) and days.
and and
in the
(10 Compound
in orally,
DA/NE
the
Daily
tested
suggest iced but
ratio
the of Tissue 80 in
day 1.5
SHRs
- maximal
6
and
DA/NE
is
heparin fold
B plasma
tissues fold h 16 0.4 or
of
in blood either
at more at were ratio after
2.7-
that
and
NE
kg, the the the
(in 30 30
M in B
2014202047 11 Apr 2014 restrained to nepicastat relative mg/kg pressure recorded the with the recorded the Hemodynamic in onset, obtaining of heating obtaining catheterized The cord and significantly The frequency The efficacy [0156] [0155] [0158] [0157]
a the atropine
orbit
new
appropriate appropriate animals animals 10.5 animals were
1
DA/NE
reaching (30,
A The Oral Oral ms pad
to
steady-state at of
at for %, for
baseline baseline maximal
-pressor on vehicle
evaluated
SHRs, 210 pulses
all (1
the in
(37 the
administration were were dosing
for respectively). affected 4 were 4 the
mg/kg,
vivo ratio
doses
study h.
eye h. and its °C)
highest measurement dose dose
pressor
lightly placed control)
of
measurements, measurements, plateau response a Nepicastat placed hemodynamic
The with that decrease 240
and of
5 and between model iv) in except 80V
of of h
nepicastat
min). attained animals ventilated
SHRs.
a after and response dose
anesthetized time in
nepicastat nepicastat in stainless
was in at
of restrainers Following
curves.
for having tubocurarine in restrainers
3-4 different
7-14 dosing. produced nepicastat points,
(30 of
observed
mean a Male its were
h. activity
blood slight to the the
resulted with
days.
mg/kg)
peak steel
pre-ganglionic high
The
and and with except
animals animals
blood SHRs then The this and
frequencies
56
a
yet significant and rod. pressure at
effect
(2
Harvard precise of (1
hemodynamic hemodynamic
sympathetic
ether the allowed frequency-pressor study, in anesthetized mg/kg; is
significant
nepicastat
mg/kg, pressure allowed (15 at
The were were a unclear
10 at
0.3
and
dose-dependent
-
reason
approximately
mg/kg the and rodent
pithing
16
(0.15, mg/kg b.i.d) to
(p nerve treated, treated,
iv), the
to
rats week of recover
< drug was decrease at
drive
left recover
the
for with dose. 0.05)
ventilator. 53
produced
parameters parameters
stimulation 0.45, rod were present. (180,
further
orally, orally, old) femoral the animals administration, ±4
response
pentobarbital, was to for
lowering
The
pithed
at loss 1.5,
210 6-7 for were mmHg antihypertensive cardiovascular
30
assessed 10 stimulated
with with
a
response
artery After days, were 30 Heart 5, of and
- significant (PNS) were were and
40 used
and
curve
anti-hypertensive 15
- either either
of
(33% 240
minutes.
40
30 pithed
then administration
and Hz)
the rate mean
in continuously in continuously respectively.
of
placed
was
min. mg/kg, was min)
electrically
conscious,
vehicle vehicle
the vein
the plateaued effects reduction to
was increase
through
tissues.
slow
arterial
shifted
effect. obtain study. spinal
and
After After were
on
(9.8 not
or or of in a 1
2014202047 11 Apr 2014 thyroid the tissues, the used of Example in a significantly fold significantly evaluate (arterial-coronary study compound animal active %, %, after deficient [0159] [0164] [0163] [0162] [0161] [0160]
known
the plasma
p p vasculature study
the as < in <
were:
inhibitor mesenteric
2. Concentrations The Since 0.05; 1. 0.01) it models a
function
the final the
7 Sprague-Dawley potent (days has positive
collected to to a)
nepicastat findings
safety
affected.
days (p< frequency-response the 4 been evaluate dose evaluate
and of h 3,
and
suppressor
heterocyclic 0.05)
post-dose, were
sinus) 7 artery DBH. control, Plasma 7 tested and on
is
and from and of
these
the day
the
These tolerability of
effectively to
this the evaluated
9). and 9,
(p
for
The probable
dopamine congestive effects the (venous)
10. rats caused
42
< findings study of
effects Both whereas
the coronary
data
compound portion 0.01 right
% (n mammalian
treatment
of and
=
doses
a suggest curve). suggest at of
mechanism
relative modulates
catecholamine
9-12) in
nepicastat significant of
and
nepicastat will nepicastat heart 44 doses
of sinus
a
various 57
of
nepicastat norepinephrine %, dose-dependent
is
for
that
of be that
nepicastat
failure also
to thyroid p The catecholamine of
CHF.
10
< the
vehicle
on
nepicastat
for the nepicastat reduction
2.0
over
devoid doses showed 0.01) days.
heart
changes
its levels
(CHF)
subject
is sympathetic function, and
12 anti-hypertensive significantly
structurally and
controls)
Methimazole of
of
weeks. rate
no
were 6.2 after
inhibits manner
in
is
patients. significant from nepicastat
T
of levels
significant serum 4 a
mg/kg, response
the (days four potent, determined a
baseline but
future drive
the effects after (maximum similar
raised weeks levels
3
The
not
(1
behavioral sympathetic po, on and
selective
four effect effects to
mg/kg, to
publication.
in in:
objectives
the
of
to of
transmyocardial b.i.d and 7,
in PNS cardiovascular the
weeks,
methimazole, nepicastat
T
DA/NE 46
942
in shift 3
12
throughout
cortex po, in (day and
% SHRs.
effects
was weeks
samples
drive
iodine-
and
b.i.d.), of and of
orally
3,
ratio
~
4 not the As
on 58 31
to to in h 5
2014202047 11 Apr 2014 NYHA resistance, mL bands with by these the patients were ventricular of of of counting. liquid/liquid Triplicate and corresponding after [0167] [0166] [0165] [0172] [0171] [0170] [0169] [0168]
plasma CHF
the a
of
day coronary
radioenzymatic
four catechol-O-methyl patients. supine,
plasma. for
incubation,
Concentrations Samples
class
over were prior
weeks using b) samples The
each c) g) d) e) end f) MV0
Blood
after extraction 12 at sinus
to collected
quantitation to
diastolic
The 2 weeks aliquots catecholamine of
2 2
Left Six-Minute Hospitalizations Hemodynamic Quality hours
, the four
hours blood the of
catheterization pulmonary
linear method.
blood
ventricular start weeks
of O-methylated
and pressure post-dose volumes transferase
of on
post-dose.
were the range of
0.045
limit
of then day were
Walk free and life
The
collected dosing) are artery
is
of parameters, 0 and
at mL
separated
ejection base 12
collected and
during method (QoL), during 1
Test the (i.e. marked
12 and
In to
weeks
heart to
catecholamines
pressures,
weeks.
changes
method of 333000 addition,
58
at 1 after tritiated the from
mL.
weeks dopamine week
fraction, rate
involves
CHF a
by
and from day
including
four time patients
thin is at pg
in
4
a then 4
S-adenosyl
prior
symptoms, four and 1
the weeks
at group of medication
and corresponding
pg
left layer the and 2
dopamine
arterial
scraped and pulmonary are of from hours
to cardiac 12. ventricular incubation
norepinephrine of
and
dopamine chromatography.
the
extracted 12
patients
a
Further
methionine.
post-dose
12 weeks
vein peripheral Global
start dosages into
output,
or weeks
norepinephrine artery to
of and
end
or scintillation underwent of samples
from
the
2
Assessments,
norepinephrine
systemic
dosing) and coronary
systolic, for hours were vein,
wedge plasma On the
on the
The from
completion determined whilst
plasma right day treatment post-dose.
at
vascular
vials pressure relevant sinus
samples
and per
a supine
0
heart
time and they
(i.e.
mL left per
for by of
2014202047 11 Apr 2014 N-acetyl nepicastat. performance was norepinephrine, phenylephrine the nepicastat norepinephrine into Example intravenous Daily inhibitors DBH activity, dopamine/norepinephrine significantly levels spontaneously affinity showed 3) [0174] [0173] [0179] [0178] [0177] [0176] [0175]
cardiovascular
R
analyzed
four inhibitor enantiomer
in treatment
Nepicastat A Preclinical In Overall, The The
for a
and the
metabolite
8
pooled
binding for
on
studies categories: these
binding
heart, The
of its aortic
administration the
human
responsiveness in
SKF-102698
hypertensive the decreased binding levels an nepicastat
receptors human
in (18 effects, receptors. singlicate
with affinity
was mesenteric
increase
method.
in
with affinity vitro contractile of
nM),
DBH
vitro
and a nepicastat
nepicastat
affinity plasma
potent
and ratio of 1) studies
male
listed was denoted
in
increase for was each
biochemical (85 norepinephrine less and
Thus
rats
resulted 4)
or to
dopamine,
in
nepicastat an
inhibitor behavioral sample
nM). at renal values response
above.
than
9 spontaneously
the day in assessed in
(SHRs)
nepicastat selected effective nM as (10
rats
vivo
mesenteric
dopamine
5.0 Compound artery, during The
in 59 comparable was (CL
mg/kg,
and
of and
for
was effects
pharmacology
relative in a S the effects.
receptors.
used
both
inhibitor
6960),
and Ml, enantiomer monkeys, and an routine
significant and vitro screened
ability
levels), increase
artery
p.o.)
hypertensive the bovine as B. Dl its
(i.e.
to to the significantly
cerebral
primary to
increased and
of use the
The of
normotensive
from in
showed Quality
the at
DBH 2)
nepicastat phenylephrine
and
in the Wistar-Kyoto D2,
of studies
(p<0.05)
selected SHRs
in
the effects 0.5
ability
human the cortex
metabolite
vivo compound and in
rats dopamine/norepinephrine Control a to
lower method
rats
similar
dopamine for 5HT
were
4 studies
receptors. was
on
in
hours DBH.
decrease to (SHRs), Wistar-Kyoto and
both 21 ia
thyroid rats. more than sample
,
is conducted decrease were to
lack to 2A
are
dogs.
following days
species.
inhibit
impaired The and
monitor
that
potent
subdivided Nepicastat of not
nepicastat and
in
(QC) function,
2c. restored
IC50 binding
Oral
for
potent
tissue tissue
DBH
with than rats.
oral
The
and the the the
for or in
2014202047 11 Apr 2014 norepinephrine mg/kg, mg/kg, ventricle potent norepinephrine ventricle the mesenteric ratio i.v. observed or in increases in Dawley catecholamines days effects cortex caused dose-response and dose (up dopamine/norepinephrine decrease cerebral significant [0180] 1 10,
hour,
dopamine
i.v. the
left 30, to injections
100 via in responsive
25
in
were administration The respectively. in a cerebral
or ventricle the and
rats
mg/kg.
left
the cortex. and
significant days) in inhibiting in the
100 and
artery mesenteric
in effects was
the increases
the
drinking at significant
mesenteric (3.0
ventricle).
left study mg/kg,
(15
dosing. 2.0
was left
mesenteric levels concentrations,
dopamine/norepinephrine long when cortex In
at norepinephrine
and
mg/kg) ventricle SHRs
of
mg/kg) ventricle and 1.0
SHRs
DBH studied dose in
The lasting nepicastat water
cerebral at and
artery assessed
in were mg/kg, SHRs,
6.2
at only artery
6.2
dosed
increase ratio dependent
in and there
tissue
dopamine
10 Significant given mg/kg were in
(0.3,
the artery. (12 of
mg/kg. observed at
at
three
male the cortex. mg/kg.
and
(1-6 in
male
doses
were
6 in and at
cerebral hours was
30
0.6,
the seen
hours
in
dopamine/norepinephrine 12 male
p.o.
in
and 1 mg/kg/d).
SHRs
doses
and
decrease the Sprague-Dawley the
mesenteric significant harvested and or the of
Significant hours
and
Taken in at
The following
b.i.d., 60
changes In SHRs following
dopamine/norepinephrine 0.3
10 dopamine/norepinephrine cortex 100
10
cerebral female
1.0 were
following an ratio
Sprague-Dawley the mg/kg, mg/kg, apart. mg/kg/d
effects
together, mg/ml).
mg/kg. with in and
six
administered in
elevation of artery changes dopamine/norepinephrine norepinephrine
Wistar
changes
in
cortex 10
the a Sprague-Dawley rats no
hours
in mesenteric oral
The single
of
mesenteric
days rats p.o.
mesenteric the significant
in nepicastat In Similar (60-100
rats
administration
only in nepicastat rats
24 after
conclusion,
left 6 in
for oral
of the
hours
in
ratio
hour dosed these rats
12 with
ventricle at
the
dosing
either artery, significant dopamine/norepinephrine
mg/kg/d) dose
hours (10
100
resulted
dopamine effects
artery (3.0 there artery after
ratio third
either time
parameters
ratio with rats
mg/kg)
mg/kg.
on
at
nepicastat 7 and to apart
mg/kg)
the
at was
of
dose. 0.3, were
were
course or at acute nepicastat observed
in dopamine dopamine
male than
in 3.0
either
24 dose-response second 3.0
ratio
25
significant at and the a 1,
In found mg/kg,
hours
Nepicastat significant significant
or
in either and in were mg/kg, 3,
days
Sprague- of
a was increase
10
cerebral
chronic the the
second in 10,
of
in tissue
or
for to at
also
two less 3.0, had and and and left left
the the the 30,
30 30 be
in at 7
2014202047 11 Apr 2014 peroxidase mg/kg/d. nepicastat that inhibitor hours or When in interest increasing increase significantly from and single dopamine/norepinephrine and levels dopamine/norepinephrine significantly extending cortex, doses days) a stimulate enantiomer artery SHRs. [0181] [0183] [0182]
decrease
the 10,
mesenteric dosed there
did
dopamine/norepinephrine in of nepicastat that apart from dose, 30,
Nepicastat Nepicastat Nepicastat to
and
triiodothyramine in
vitro SKF-
was not
the
through assess
for was
in and dopamine
in lower
in dopamine male
(30 renal or
plasma less vivo, alter release
no the 10
decreased 102698 artery a 60 three
studies
mg/kg whether
was
days,
potent, significant
beagle
tissue left
has caused 60 medulla, effects mg/kg/d) plasma (the nepicastat
renin
doses
levels mg/kg/d. given resulting of
ventricle
structural
and while
in
using
p.o.).
norepinephrine
renin dogs
S
orally
decreasing ratio a or
decreasing than ratio renin
activity. the
at at to significant
enantiomer) in
increase with thyroxine there methimazole
30 male 5 at
purified
in from and the
dopamine/norepinephrine nepicastat and
active ratio, Similar 30
SKF-102698 similarities hours activity
significantly
doses mg/kg. the
a
was
cerebral mesenteric increase
beagle
plateau norepinephrine these However,
in and left
but
norepinephrine
inhibitor in after decrease
of 61 a
dopamine. results
DBH
in
levels,
male ventricle
significant did was 4 in was The
(2
dogs
male
60
or to cortex. administration
plasma vivo in
mg/kg/d)
not
in increased methimazole,
(see were Sprague-Dawley 12.4 nepicastat potency
artery significantly
significantly
response
of in
does
for mg/kg/d, SHRs.
alter decreasing
studies
norepinephrine
and DBH
In
mg/kg/d,
levels 4.5 above). observed Norepinephrine renin decrease
in
not the
and dopamine
significantly
mesenteric
relationships
days
SHRs
Thus,
in at
strongly
ratio (30 cerebral beginning
alter with
increasing activity.
of
dogs
and
more all
a p.o.
more
(5,
norepinephrine and
in
30
in potent However,
after
nepicastat, in nepicastat
plasma
rats
the doses. levels,
15,
norepinephrine, mg/kg
had
at 100 parallel the cortex,
levels artery potent
dopamine
reduced three potent
fed
left and doses
at It in
has
inhibitor no dopamine mg/kg/d renal
was in renin the
a 10 ventricle,
intraduodenally. 30
in
effect and
been
nepicastat norepinephrine doses In would low
than those the
when
of
mg/kg/d
SHRs left mg/kg then serum artery,
therefore
conclusion,
an levels mesenteric at iodine activity
of
p.o. shown the and
on
ventricle given obtained
increase result given
least and
thyroid and
the
except
after
serum
levels b.i.d., DBH
renal for
diet had and and
the the
an 10 12 of to in in
at R 5 a
2014202047 11 Apr 2014 telemetry were nepicastat(10 pressure nepicastat mg/kg responses though nepicastat pressure made of i.v.) dosing antagonist (10 and conscious, serum alter despite did calculated flow sympathetic effects [0184] [0188] [0187] [0186] [0185]
triiodothyramine
not
mg/kg, 30
decreased heart
restrained or
using
levels
in in Nepicastat Nepicastat Nepicastat Acute Daily mg/kg). causing impair the p.o.).
when was
alter
SHRs
conscious,
blood rate, to
were renal SCH-23390. restrained causes
p.o.) antihypertensive the
nerve
of
treatment preganglionic less
mg/kg, intravenous
renal
urine mean
given triiodothyramine or and a
Thus, do DA-1 mediated The
were pressure vascular
decrease
systolic significantly stimulation. (-13 a induced not
their also
function
or restrained
decrease
arterial antihypertensive
production
SHRs alone, p.o.)
not
antagonist
appear nepicastat
with thyroxine. mmHg)
nepicastat blood
significantly transducers resistance attenuated blood in by
treatment
induced (1.0-30
a nepicastat thus nerve
pressure effect arterial in
to DA-1
in significant
normotensive pressure lowered anesthetized
than
be
or pressure
making
blood SCH-23390 or
reduces
stimulation thyroxine. mediated
is (10 Thus,
mg/kg,
blood receptors. a was urinary by
of with
over
more
at effects
(1 significant
mg/kg) 62
measured blood attenuated pretreatment pressure anesthetized
doses and the as
decreased nepicastat, antihypertensive
pressure. SHRs a
the p.o.),
pronounced
measured
SHRs,
3
via
Wistar-Kyoto results of excretion
10 to
pressure
hour in of also
rise
nepicastat
However
mg/kg, DA-1 assess
(-46 and in
directly
pithed 30 antihypertensive
and
period, reduced the
following
both uninterpretable.
in with unlike SHRs significantly
and
mmHg). by
receptors.
in
if did of p.o.) blood in
hypertensive
the SHRs
SHRs the via
this SHRs the
100
in
rats; not but sodium SHRs. effect
with methimazole,
blood
tail in
conscious, an renal
dopamine
mg/kg/d compound
dosing. decrease
did pressure SHRs
however, instrumented 5 To arterial
cuff
and
nepicastat
for
reduced hours pressure
not vasodilator The effect or summarize
method. normotensive Overall,
for
up lower
potassium.
and
for cannulae. renal An restrained antihypertensive receptor
in
after
21
the reduced to
when did
heart
4 30
attempt response days
(3.0
tachycardic renal 4
with decrease
blood hours
nepicastat not
dosing However, effects days,
together, hours
rate the
(DA-1) mg/kg, did
radio SHRs blood blood affect
after flow rats,
was The rats
(10
but not
of (3 to in in
2014202047 11 Apr 2014 p.o.) produced norepinephrine norepinephrine nepicastat nepicastat the occur of in in observed output observer instrumented combination sympathetically-evoked dopamine decrease dose arterial decrease additional effect significant [0191] [0190] [0189] 100 100
arterial anesthetized
the
peak mg/kg/d mg/kg/d
(3-30 that
with
over
or antihypertensive Nepicastat The Studies blood
in in in
decrease
in renal failed
no
blood
blood had
levels, mg/kg reduction
following
enalapril 30
renal blood SHRs
blood chronically
of significant
resulted persisted pressure,
with
a days.
blood
male a
levels to
levels pressure in greater pressure
blood dose
i.v.).
in
pressure lower at in (30 pressure radio-telemetry
normal
alone.
the in
Heart beagle doses
blood in flow
acute
were of over
and left
mg/kg left blood flow
functional effects instrumented, effects a
blood mesenteric in the
decreases decrease
for ventricular
rate was ventricular of response
Thus, a an pressure significantly
intravenous dogs. anesthetized lowering
in angiotensin
24-hour
pressure
up
30
intraduodenally) pressure were increase response was of -10
to
and 30
responses.
nepicastat
in along
blood However,
mmHg,
not five artery observed to days was
blood period
pressures 100 conscious effect lowering mass.
63
carotid
with
increased,
in to hours
converting dosing
reduced, with
-17 of pressure
dogs mg/kg/d
5
direct the
pressure
compared
treatment after
nepicastat of hours A over
nepicastat mmHg,
at a
artery
after
reduction (including effect reduction
dopamine/norepinephrine nepicastat did (1-10 showed 3 dogs
renal
a
there and and
30
transducers
for single after dosing. enzyme
not and, occlusion
in
studied compared days
of to motor 10 mg/kg nerve 7
(30
was
caused
SHRs
significantly SHRs days. dosing. -20
when in no
in mg/kg/d.
dose, peak
in mg/kg)
of
left inhibitor A left
no
activity stimulation, in cardiovascular
for
normotensive
i.v.) than dosing,
up similar
was with combined dp/dt),
a ventricular At and ventricular
SHRs. significant to
significant 12 to
Thus,
caused 30 in -42
with
The there less nepicastat 5
hours enalapril was
normotensive
hours lack mg/kg/d inhibit heart and in
At effect
although than
no
or with was unaffected. ratio,
SHRs.
a
after mass. of resulted mass
inhibition 100
rate,
the potentiation after Wistar decrease changes
effects effect
either the no (1
at enalapril, at
the mg/kg/d increase
a but
cardiac did mg/kg,
30
30
loss dosing single tissue effect Thus,
peak
rats. in was
rats and and
not not the
of of of in in A a
2014202047 11 Apr 2014 treatment was that mg/kg/d ventricle mice increases. increases hypertensive observed in or a challenge, significantly another and dosing clonidine auditory and following for effect SKF-102698 [0192] [0195] [0194] [0193] 10
maximal
central acoustic
mg/kg/d
DBH
modulates dogs
no 100 (10-100 on
Nepicastat in No Nepicastat When
statistically
DBH
p.o.)
startle
and sympathetically-evoked mg/kg/d, inhibition with nervous in SHRs (20 significantly in
acute
behavioral Thus, but startle
decrease
vivo. in
response
mesenteric lowered response
nepicastat mg/kg (100
mg/kg, for
nepicastat the inhibitor, the
produced reflex (3-100
dosing
chronic
reactivity
30
system
is action In
p.o.. caused heart,
in mg/kg/d,
significant
i.p.). a days in
rats,
effects b.i.d.,
the
blood
to were potent reduced to mg/kg/d,
at
tissue
was
artery
in
dosing of
SKF-102698, mediated
Rectal renal
carotid heart only sympathetic in 1.0-30 oral
Acute
no
rats. the (3-100
p.o.).
significantly were given pressure SHRs.
p.o.), competitive
norepinephrine relative treatment sympathetic
significant and slight decrease the artery
with
p.o.).
temperature In functional mg/kg,
artery administration
observed
mg/kg behavioral
to
increase Motor mesenteric dogs,
and
In nepicastat
male
and and
throughout to nerve 64 Thus,
nepicastat
conclusion,
occlusion
in p.o., with the
i.p.). by
reduced
a
inhibitor kidney. activity
non-significant
beagle effects
plateau responses. nervous the in
in
cerebral
nepicastat the
stimulation was
effects and
nepicastat rats levels,
heart artery at degree
to
centrally a the by dogs
it following also showed was
in
effect rats on of dose nepicastat Nepicastat
system.
rate did
in
at cortex. treatment
does
anesthetized
human day
of rats. gross unaffected. also did for did a
not
that resulted in for
in
dose
blood
inhibition
when 4.5
not not some not
acting response
unaffected
rats effect 10 DBH has
No
motor
DBH
is
6
significantly days cause effect have
days
with
mg/kg/d. pressure a been behavioral (3 in
dosed selectivity
potent inhibition locomotor
animals.
a-adrenergic
significant
in mg/kg
at Motor of
the to a behavior detectable of locomotor
shown
vitro,
10 major
dosing over
an
once
blood
DBH DBH and mg/kg/d
In
i.v.
p.o.),
activity effects
reduced
occurred
30 to and
to
Nepicastat contrast activity
inhibitory heart
daily
at
tyramine evidence
inhibitor inhibitor in pressure
result changes the
days activity
agonist 10,
in and
there mice were
rate rats and (30 left
30, the
of to in in at it
2014202047 11 Apr 2014 nepicastat with potassium ml) methanol were with ml/kg. hydroxypropyl higher Example Detection Hypersil dosing and analog - (free from acetonitrile. aliquots containing dithiothreitol). and and dose Tween [0197] [0196] [0199] [0198]
2.0
was
the
and
to g). base) solutions known heparinized surgically a
oral standard Aliquots Male Brains The
and At
80
of
determine
higher
verified of Internal
BDS 9 extracted
was
in phosphate, various
(all containing per
analytes nepicastat),
doses the until
studies
brain. Solvent concentrations rats
were
was by ml.
obtained 15
methylcellulose
Samples excised,
aqueous concentrations curve
of
by
standard 4 (Crl: of
UV syringes, cm
times
5,
penetration plasma with hr rinsed
was nepicastat, described liquid
composition
Cs 15, pH
absorption generated
after CD 5
and and from
were after phase
back column 3 and
3.0,
pg/ml
was
briefly
chromatography BR
(0.05 ml
and
dosing. of all
brains
50 dosing, with mixed Sigma
extracted
here added
Vaf+) were
analyte. to were of samples
(50 of
plasma
mg/ml
from
at
at
or a with was
compare
ethyl
5
nepicastat
261 ambient
centipoises were
0.5 monofluoro were
with
mM Chemical a (50 Doses assayed samples
weighing
the 40% saline,
translucent
65
for
nm. were ml) was
with
Plasma acetate/hexane pl
dodecanesulfonic 200 designed analysis
homogenized
the
the
B
(LC).
of
were temperature. were
prepared Concentrations
frozen blotted and by 250 of
mM methanol
into
pharmacokinetics 10, Company). viscosity), 180
blood
concentration
LC.
analog was
mixed formulated
The
30, pl of
suspension. sodium
to the - at
on plasma of
220
and
pumped by -20°C were 5 evaluate The
containing
250 CNS a
mg/ml (1/1, with
in
of paper g Mobile centrifugation. 100 phosphate
1%
acid LC
Concentration
were obtained
of 5
mM
from until
in nepicastat,
internal
v/v).
by mg/kg at benzyl
ml data analytes
dose the system
towel, Dose and water
in
a fasted
phase acetic analysis. quantitating untreated
flow 20 of male
pharmacokinetics
are mobile
was by
buffer,
The doses,
pg/ml volumes alcohol, standard 200
then containing
used rate were
overnight A cardiac
expressed acid
and
a
and
organic
Brains of was
clear
weighed mM
respectively, phase rats of
a pH
female a
drug determined and
monofluro
and 5
1 Keystone levels
(50 were 12.5
puncture
7.0, fortified solution ml/min.
sodium
of
mg/ml before 100 2% B
in phase as 0.6%
pl
rats,
mM was
(1.5 (0.5 rats
2.0 the
pg of of of pl 1-
2014202047 11 Apr 2014 were phosphate, between nepicastat terminal Plasma Levels increasing are emulsion), gently a (expressed 77% 30 following elimination single concentration. as: calculated (AUC) determined [0201] [0200] [0205] [0204] [0203] [0202]
30
mg/kg expressed
mg/kg higher extracted
back oral
of AUCtotai Pharmacokinetic Following Plasma Plasma Concentrations half-life
from linear 10
nepicastat
the
in
oral by
pH as doses 100 dose, oral by half-life and
extracted
brain
in
the as
10, pg/ml). zero
linear levels portion 7.0, levels dose
with
μΐ
dose. female 100
= (T1/2) pg
were trapezoidal
30, and addition aliquots exceeded
AUC
containing
to mg/kg, (free in appeared
of
10
regression with
of and of
was
brain the of of
From obtained. the nepicastat
rats ml
nepicastat parameters nepicastat
base) (O-Ci the nepicastat
100
150 calculated
were of relationship
of based time
those
(expressed than
rule.
2
data.
to
100 mg/kg as ethyl
μΐ 0.5 per
hr
t)
of increase assayed
of
of
on in in
Concentrations
to +
following μΐ mg/ml were g the AUC
in
Areas
250
acetate
plasma. were
in
of values the male Ciast female
as of oral
male
log
66
brain plasma
as
0.693/β, mM methanol higher between
/β by
from slightly
last
dithiothreitol. doses under calculated pg/g) plasma
rats
of rats
/
where
LC
rats,
dosing tissue. acetic hexane
AUCtotai. quantifiable
zero in of
given
were increased as to
the
where
at
of AUCtotai female the
male
to concentration
male
described acid.
Ci
to higher
onward, plasma nepicastat
the
from (1/1, ast
initially plasma an infinity
β rats
rats, is
aqueous Aliquots rats
is
equivalent
linearly v/v). doses mean
and plasma the the concentration
given
however,
for than respectively).
AUC lower
(AUCtotai
in last
elimination
dose vs.
plasma. (1.70, The
plasma
phase
plasma of
in with to 10,
concentrations
than tai time
quantifiable
male dose homogenate
organic
was
30, of
concentrations
2.09, increasing ) (to
those
data concentrations.
was nepicastat increased
vs. Level rats or
of
linear. rate disperse
Following
and
time 100 nepicastat. phase
within calculated in following
constant in
3.88 plasma plasma
mg/kg
(2
curve doses
brain were
with
was was
The any ml) the
of hr
a
2014202047 11 Apr 2014 were At perchloric rapidly prepared randomly of Example of Both in hours. oral 400 following levels analysis carcass anesthetized and groups either artery [0207] [0206] [0211] [0210] [0209] [0208] 10
vehicle nepicastat nepicastat
the ml/kg.
24
administration grams tissues measured
nepicastat
of catecholamine
Nepicastat Animals Following Each The (n=9 Sixteen- into time
hours harvested
of nepicastat the
10
(dH assigned, a
acid
variance purpose
a
were were per All single with morning treatment
of (10
in tube 2 following
O)
17 at
in were brain group): decapitation, (10
doses mg/kg)
halothane,
immediately administration to was
1, allowed containing week
and a oral in
of
of mg/kg)
the yield 2,
(ANOVA) centrifuge levels
of dosed brain
were
vehicle synthesized this
group
4, weighed. of
dose
administration. afternoon old, a on
administration,
6,
an single
nepicastat study
were
were food
every initially or dopamine
decapitated,
8,
oral male heparin, in
was
plasma
(10 vehicle
frozen
12, tube
with
spontaneously of greater was determined
oral
and
dose minute
compared ml/kg) as spontaneous before The 16, a
lower
and
10 to effects the administration
in and were
water samples (dH and
that and 67
mesenteric determine than mg/kg liquid
and 9
hydrochloride the
the
to 2 centrifuging the
norepinephrine
than O; treated could 24
administered
using
the be TRT, morning in to ad
left
10
study, hours nitrogen
oral
were hypertensive plasma. hypertensive
vehicle those sacrificed
left
libitum.
ml/kg). be
ventricle
the HPLC
HARVEST animals
dose
artery
of administered ventricle after taken
to of
24 in
at
nepicastat
and salt
at
one sacrifice. of
plasma, 4°C. hours
with
a as levels
at each was
Animals
nepicastat put by
and stored and
single
rats.
of rats f, free and
electrochemical
time and draining
into put nepicastat 2,
time the
9
in
but
at (SHRs) in
mesenteric at base vehicle oral At 4,
Catecholamine
the
their 10
repeated course in
following
an were -70°
point. 6, from to 1,
mg/kg mesenteric
administration
0.5
empty
2, equivalents 8, male blood
interaction
weighing
C.
was
4,
animals weighed 2 12, of ml
A
artery volumes
6, hr Mesenteric
rats,
or the
detection.
treatment dissolved cryotube. 16,
from two of 8,
onward, a
effects
12, single levels artery levels
or 0.4M
were were
300
way was and and
the
24 16 of of
2014202047 11 Apr 2014 Nepicastat treated were values performed. to vehicle vehicle norepinephrine out Pairwise + intravenous Example in However, dopamine/norepinephrine significantly allowed dopamine and administration across spontaneously afternoon [0215] [0214] [0213] [0212]
the any 25%
using norepinephrine
marginally
first
of most were
Nepicastat animals In The Sixteen (1.0 (75%
DMSO;
food
the analyses
11
Fisher's
general, before levels
administration. a was
ml/kg) (iv) of A significantly three purpose
consistent (p<0.05)
and
propylene
of
one the hypertensive to at or
administered
were administrations,
the nepicastat (0.05 LSD the 1.0 17 water between parameters dopamine nepicastat first way or was in week study, 1, measured nepicastat ml/kg) higher of the strategy f2 2, increase ANOVA ad glycol synthesized (p<0.05) ratio this 4, hours old left treated libitum. (hereafter to rats 6 levels as had than were or male and ventricle one lower to was + the study six at of at 12 in 15 few with 25% control those lower and 15 12 observed. following hours of treatment. free Sprague-Dawley 1, hours the significantly mg/kg Animals mg/kg. and hour statistically referred at the 2, was factor vehicle DMSO) 68 base in of dopamine/norepinephrine the than after the 4, following apart, was time Sprague-Dawley vehicle 6, to of 6 experiment-wise equivalent TRT a vehicle were to the At hour animals, 8, single dissolved points. nepicastat. to determine of as (p<0.05) significant the last 12, obtain was at either nepicastat) weighed time treatment rats, only the f6 oral compound 16 performed and at and or 2 vehicle a weighing point. in administration at each greater dosing and the prepared rats. 24 effects and Tissue the the 24 error groups on time hours 6 harvest effects 4 administration. randomly appropriate Dopamine (75% the hr Animals for than ratios hr volume rate. 300-400 on norepinephrine harvest point, time the levels each (n=10 following mesenteric propylene those time Norepinephrine of were afternoon at point. of received harvest assigned, were of 10 grams, levels times. intravenous per no amount of 1.0 dopamine mg/kg observed changes vehicle group): dosing. carried Levels ml/kg. glycol artery time. were prior were The two and the of in 2014202047 11 Apr 2014 Nepicastat perchloric was was the norepinephrine pairwise performed ratio Example inhibition 472%, overall Fisher's liquid Tissue administration active enantiomer at spontaneously dopamine compared [0217] [0216] [0222] [0221] [0220] [0219] [0218] 3, effects rapidly 10, repeated primarily chromatography in and Nepicastat A dopamine Each In Compounds This This experiment-wise LSD 30 rats. comparisons 12 one-way of to and conclusion, acid. of for was significantly or (Compound study vehicle DBH harvested rat study nepicastat test. 100 norepinephrine. hypertensive norepinephrine rats 12 levels dissolved due was Tissues and also mg/kg in A analysis were hours assessed administered treated to were the dosed Bonferroni by norepinephrine between using compared intravenous and heterogeneous with (p<0.0 B) type left were anesthetized p.o.. 51%, in later prepared following rats animals. iv weighed. of ventricle the SKF- electrochemical the 1 in immediately in 1) variance error nepicastat A and (SHRs). appropriate the the increased the the adjustment effectiveness Kruskal-Wallis 102698, at administration significantly rate cortex, efficacy tail following three of and concentrations with variances The 69 15 (ANOVA) Sprague-Dawley of vein Animals treated a administered the 5%. frozen doses halothane, ventricle mg/kg DBH amount left was detection. of the dopamine/norepinephrine of the ventricle, morning. (p<0.01) among performed rats (30 was inhibitor afternoon were in with of nepicastat S of liquid significantly were mg/kg). was and enantiomer performed decapitated, nepicastat vehicle given a treatment rats. as vehicle main placed and increased assayed previously nitrogen before on Six the three in mesenteric This (dH all effect for resulted hours (nepicastat) free altering in were p-values 2 by and groups. doses, harvest. (p<0.01) study O and dopamine dopamine 1.0 shown for high for base the performed ratio stored after ml also nepicastat artery treatment 12 the in left performance to iced Subsequent to The hours equivalent. by with significant compared decreased ensure the levels levels at and be ventricle of 1117%, dosing -70°C. 0.4 orally the using apart male their final was and by an M of R 2014202047 11 Apr 2014 randomly nepicastat, were nepicastat using were rats performance weighed, morning. River PEG Einear Dose dosed distilled (dH greater, as compared second Overall control 70°C. decapitated, [0223] [0227] [0226] [0225] [0224] follows: 2 treated O), used, 400:dH carried and orally Eabs) a Models. Fifteen Four The This Change animals. statistical results water, the Tissue or nonparametric placed nepicastat assigned at blocking and a with At nepicastat dopamine new and 2 series norepinephrine doses liquid linear (p.o., were 0, out to 30 are six = or various dopamine 50:50 in the variable (30 sixteen A using mg/kg test 3) reported. of using of chromatography contrast hours allowed to iced nonparametric cortex, to factor PEG mg/kg 30 statistical one compared at concentration vol:vol SKF-102698 Dunnett doses 0.4 t-test. mg/kg week SKF-102698 a 3, was after 400:dH of gavage and Day was - mesenteric M food 10, concentration Pairwise Vehicle) the of tested old for perchloric using the analyses developed ’ norepinephrine s was 30, The nepicastat, SKF-102698 2 following and male 0 SKF-102698. test needle) one-way and third vehicle at and for in N a performed were third analysis water EPicASTAT 70 nonparametric to doses artery, electrochemical spontaneously the equality were 100 acid, dose control which three prepared was and treatment statistical cerebral analysis or of ad mg/kg, between and performed. to frozen SKF-102698 rats - Compound 30 significantly concentrations times libitum. to for Doses calculates (30 the the left mg/kg. zero in were mg/kg each of cortex t-test. PEG-dH 2) hypertensive groups: experiment-wise in test ventricle 10.0 treated 12 detection. of variance by liquid Compound The hours anesthetized tissue Animals 3, compared B ml/kg the Since - the was A at Vehicle) (p<0.05) at 10, 2 1) and first O 30 fourth nitrogen, difference were SAS were 30 apart, (ANOVA) distilled 30, and significantly two dosing vehicle mg/kg. controls rats mg/kg series were B procedure and rapidly statistical S assayed Compound different strain KF-io2698 beginning (SHRs) lower), error with at volumes. and of treated water 100 compared to weighed Each 30 at with differences the separately. rate. stored halothane, harvested, each mg/kg mg/kg (p<0.05) by (Charles vehicles analysis and General rat vehicle vehicle in group factor B dose high was The and at the the the of to in - 2014202047 11 Apr 2014 vehicle than not nepicastat the in dopamine/norepinephrine significantly and dopamine/norepinephrine dopamine dopamine effective did and (p<0.01) The concentration (p<0.05) significant significant greater SKF-102698 [0229] [0228] [0233] [0232] [0231] [0230] 102698 a significantly dopamine/norepinephrine the not 100 significant dopamine/norepinephrine vehicle Dopamine Dopamine In In In than Comparing at dopamine/norepinephrine mg/kg than significantly than (p<0.01) greater the the the doses and compared concentration increase increase vehicle at (p<0.05) at mesenteric for cerebral left was the Compound of increase of doses a lower than nepicastat ventricle, nepicastat. dopamine/norepinephrine dose concentration not 3, than concentration nepicastat in in at to 10, lower all dopamine cortex, of both significantly norepinephrine vehicle Compound lower of in Compound in 30, artery, doses 3, ratio ratios B 30 dopamine the relative dopamine at and 10, norepinephrine (p<0.01). ratio. mg/kg. relative than ratios with in lowering at cortex of significantly relative 30 and 100 in ratio. the in nepicastat. doses B vehicle less and the to B Norepinephrine SKF- in the the mg/kg (p<0.01). left and to was and levels. treatment at The 71 the left Norepinephrine (p>0.05) to treatment of 100 norepinephrine dopamine/norepinephrine ventricle mesenteric lowering the 102698 norepinephrine significantly treatment mesenteric increase ratio. 3, of at ventricle levels. mg/kg. dopamine/norepinephrine (p<0.05) Nepicastat nepicastat. 10, doses with than was with at norepinephrine 30 above levels with artery Nepicastat 30 vehicle, Norepinephrine was of greater vehicle and significantly artery vehicle, greater levels, concentration was mg/kg 10, vehicle, (p<0.01), significantly vehicle were 100 of were significantly 30 than Compound at (p<0.01) and mg/kg. SHRs Compound significantly in was 10, and ratio vehicle levels, Compound significantly was (p<0.05) and increasing the 30, significantly concentration 100 ratio was was (p<0.01), greater had (p<0.05) cerebral than and Norepinephrine B and more at mg/kg. B (p<0.01), resulted significantly significantly no greater doses 100 vehicle lower resulted B increasing dopamine for effect (p<0.05) effective resulted but mg/kg. greater cortex, of SKF- more with than was The in but did for on 30 in a 2014202047 11 Apr 2014 NEPICASTAT vehicle than than vehicle potent the than vehicle than than than Example Furthermore, of significantly lower and administered left dopamine/norepinephrine significantly lower dopamine/norepinephrine lowered for [0234] [0237] [0236] [0235] three mesenteric ventricle, nepicastat methimazole for for vehicle vehicle for vehicle than Nepicastat In The The than was than than with SKF-102698. SKF-102698 oral norepinephrine SKF-102698 13 conclusion, vehicle dopamine greater dopamine for for for doses over lowered nepicastat the (p<0.01) vehicle but SKF-102698 for artery, than (p<0.01). SKF-102698, SK-F SKF-102698, less were 24 R was administered for for SKF-102698, hours. for (p<0.05). enantiomer the left concentration norepinephrine 102698, effective with prepared (p<0.01). SKF-102698 concentration SKF-102698, greater was dissolved SKF-102698. data relative ventricle, ratios ratios treatment, more The SKF-102698 show and and and than in and in 12 in dopamine/norepinephrine effective Norepinephrine (Compound the to in and the the the the hours that in and than administered the vehicle and vehicle in vehicle cerebral relative the increase increase increase however 72 left the nepicastat cerebral Norepinephrine the for left the apart. left than ventricle treatment, nepicastat mesenteric increase for decrease (66.7% ventricle more ventricle to cortex, B) above above SKF-102698 above treatment cortex The vehicle SKF-102698, as concentration is in the than a were propylene S following vehicle vehicle vehicle was potent above (p<0.01). all was and of enantiomer, were free artery concentration more SKF-102698 SHRs significantly three with greater significantly ratios base treatment in inhibitor significantly vehicle was was was was the than and three glycol:33.3% was nepicastat six tissues equivalent. for greater greater mesenteric greater in nepicastat significantly hours the SKF-102698. not doses of SKF-102698 the was was (p<0.05) with (p<0.01). greater DBH increase at different after for for for cortex greater significantly significantly at greater 30 Nepicastat nepicastat nepicastat artery nepicastat nepicastat dH2O) was 30 in the (p<0.01) vivo mg/kg. greater greater mg/kg above more were from third than than The The and for to in 2014202047 11 Apr 2014 the treatment prior yield mg/kg. ANOVA radioimmunoassay. weighed. Methimazole if of of halothane, diet administered at (p>0.05) experiment-wise controls experiment-wise each calculated cortex, days. for served [0238] [0242] [0241] [0240] [0239] a variance the norepinephrine evening significant dosing). (Purina, to treatment dosing 2.0 Norepinephrine as At Male To Orbital and treatment. Norepinephrine at different positive with groups Tissue from decapitated, statistically and four (ANOVA) striatum and each Sprague-Dawley 5891C, in at blood solutions difference 6.2 factor a the hours the Serum 1 group error error 1.0 samples (n=12 mg/kg, dose controls and Animals in mg/kg day following were Lot ml/kg samples the rate. DOSE was evaluate rate. after dopamine and were -3 levels were levels per samples of from 1478, immediately or were cortex time dose conducted. for the volume. were the vehicle group): For appropriate were carried rats, was morning, carried determination in in cortex, control point. changes not 0.066 groups, second levels weighed the statistical compared the were weighing performed. taken harvested at mesenteric nepicastat ± nepicastat out striatum, out A occurred. placed Also 73 1 the approximately in 0.042 and dose, non-parametric at ml/kg. analyzed concentrations and using using T same analysis day to a 3 180-200 norepinephrine of in from one-way and randomly mg on vehicle and thyroid -3, Pairwise 0.4 artery day at treated Pairwise Each Fisher's Fisher's T iodine/kg day 0, the mesenteric 2.0 4 M of for using levels, grams, 3, 12 were group ANOVA iced methimazole catecholamine control function. mg/kg, 10, 7, two-way assigned animals T hours analyses so LSD analyses LSD HPLC. and 3 rats perchloric significantly a were sample) levels of and that change artery 9 nepicastat at apart, rats strategy strategy was were (day to were within The fed doses between all between in T one groups was performed 4 were ad an for 0 from the mesenteric levels, acid anesthetized doses not was levels of libitum subject iodine of dosed (p<0.05) to to 10 striatum at harvested the and baseline significantly as controls 2.0 the consecutive treated control control 6.2 a following could they analyzed using deficient to orally one-way first analysis and 14 mg/kg, artery, detect lower were days with only was and and and day the the 6.2 be in a 2014202047 11 Apr 2014 mg/kg vehicle vehicle marginally received Adult to marginally total ratio Example observed observed. (p<0.05) and dosed concentrations cerebral dopamine dopamine/norepinephrine significantly level dopamine 7, significantly [0244] [0243] [0247] [0246] [0245] vehicle. compared 30 T4 of was male by nepicastat Nepicastat Neither Dopamine The This control control. levels mg/kg cortex the 2 higher 14 oral in observed or β doses (p<0.10) (p<0.10) beagle In In -hydroxylase dopamine/norepinephrine mesenteric dopamine the to (p<0.05) study (p<0.05) in administration the the or at 2.0 in placed vehicle than the norepinephrine were daily, either striatum kidney levels (2.0 striatum, or dogs mesenteric in rat was lower vehicle lower 6.2 lowered the not in or serum. lower levels. morning control. the were artery in mg/kg performed in medulla single 6.2 following on ratios, significantly 6.2 all 2.0 a in control. vivo, with day marginally randomly mg/kg) three norepinephrine mg/kg T or at both artery, A Thus levels, nepicastat 3 and T capsules. nine. dose 6.2 both levels nepicastat. 4 compared with and tissues levels the 10 afternoon mg/kg to dose nepicastat did both ratio of or 74 2.0 kidney (p>0.05) days assigned greater on 2.0 determine significant 1.0 dopamine/norepinephrine not of affected were group, and all Vehicle 2.0 dose of of mg/kg and the to Nepicastat cause levels treatment cortex (8-10 dosing the 6.2 effect not and different appears methimazole vehicle, to group 6.2 but thyroid cortex of four mg/kg the (p any was 6.2 hours and mg/kg no from in in Methimazole, of groups days significant dopamine Sprague-Dawley was to mg/kg the other an from nepicastat. with significantly function and nepicastat apart) dogs be empty dose mesenteric decrease delivered and treated vehicle significant an striatum of no of (p>0.05) dosed for T 8 effective groups, ratio by 4 nepicastat (p>0.05) and capsule. significant the dogs levels four animals were altering in control, in (p<0.05) positive with when at artery norepinephrine norepinephrine rats. doses per different days. changes compared at significantly 2.0 inhibitor changes nepicastat. were group day Each compared free while produced than of changes and control, On higher 3 T 5, were from only dog 3 and and the the 6.2 the 15 of or to in 2014202047 11 Apr 2014 An the tissue plasma mg/kg/day). were norepinephrine ng/mL harvested, of Example stored six fifth sodium deemed catecholamines containing apart). groups daily deviations (D/NE) expressed separation (DHBA) after [0248] [0251] [0250] [0249] plasma hours homogenates aliquot day, the used at was at and Male Each To Tissue was Daily ~ pentobarbital (n necessary were AM -70° 15 as after each 10.00 in for weighed, using as homogenized heparin levels a of quantitate = separated internal analyte linear Beagle the dose, blood C. Each each each obtained samples 8/group): pg the dog and and AM. study. electrochemical were of of to last range analyte and a supernatant received the animal the samples placed determination nepicastat dogs final standard. analyze analyte (40 Animals concentrations and were dose. centrifuged other glutathione ratio for The placebo by of mg/kg, blood (Marshall divided and received 2.0 each in also brief (10 animals a for of Samples tissue per cold single were D/NE was ng/mL and dopamine The sample analysis ml) taken dog. detection. iv), (empty sonication into was gram at and 0.4 removed catecholamines catecholamines. randomly farms, were 2 extract were ratio of dose 13,000 placed of to from two doses normalized 75 centrifuged M In (10 norepinephrine of 400 of kidney capsule), drawn were concentration allowed perchloric in addition, North nepicastat samples, ml) the tissue. from in rpm The and on ng/mL the daily, assigned 0.4 dogs provided was medulla a 6 morning spiked method in Rose, each necropsy to h water M at at morning or a taken. for the at one after the acid, The - The microfuge . perchloric a the 4°C to nepicastat each sample NY) later to with (NE) calculated for weight and ad for has the one and blood end concentrations frozen norepinephrine Dogs within table libitum each the analyte. kidney point. and weighing AM 3,4-dihydroxybenzylamine a of the and of was quantitation of measurement for the acid. was treatment the were and afternoon in dose dogs at the lh dopamine means and 30 subjected On following cortex liquid study of 2 collected euthanized tissue anesthetized between After for minutes were given day collection. mg/kg of concentration in measurement and group. nitrogen were (8-10 limit 15, sample sonication, euthanized case dopamine, food of (D), treatment to standard in b.i.d at 9-16 6 rapidly plasma with HPLC of it hours hours 4° tubes once each with was The and and 2.0 kg C. (4 a 2014202047 11 Apr 2014 was biopsy ventricle. was period with within mean within variance Helmert In PROC analyzed frozen abdominal second confidence animal. steady concentration a (cf. statement, groups day 0; The [0252] [0254] [0253] smoothly particular, ln tested, SAS and BQL determined the square (0) in dog treatment is MIXED) was Plasma The The state injection varies contrast the stabilizing average defined Univariate liquid by was PROC (below which The comparisons, incision and intervals changing time taken. Box-Cox treatment HPLC for period the during set from its skull nitrogen by norepinephrine its with being of period is contrasts to GLM quantitation of correctly to Normal HPLC error statistically Tissue pentobarbital transformation; using was start the was missing. built statistics was day process, the the transformations was a manual). treatment of and performed. random and term, at day steady calculated opened using electrochemical to on theory comparing samples takes steady the included, stored require the day. and limit) as on while The first significant. electrochemical into (NE), state factor. appears p-value mean (80 to treatment the means state These at were analysis hence time in the Contrasts expose the for account Biopsies since - mg/kg,iv). the period slopes the indicated 70°C 76 was slope, dopamine error comparisons For each point weighed, might was transformations of treatment DA all detection. the However, categorical the the was calculated the until were also mean analyses the calculated. concentration and dog were following were difference be detection. frontal time fixed performed that error A analyzed. was placed the the then (DA) individually, square. group rapid taken calculated used points Plasma the since effects, were hypothesis case terms calculated. using variables lobe calculated, the This compare on to bilateral between logarithm and to from here, this performed using of following. of maximum the establish for iced the concentration the slope dog the epinephrine method the drug using each being yielding the Helmert interaction of 0.4 a transthoracotomy the cerebral The each 1 with renal mixed analysis was slope slope at group particular M steady on fixed the drug time day can slope The treatment perchloric Normal the an transformation artery one of equaling model CONTRAST of 10 cortex uses fail at and between (EPI) was and steady state appropriate the log-values. during nepicastat was which slope to contrast. and placebo the the used analyte theory (using are detect and mean set were acid, state zero dog dog and left per the the the all as to a 2014202047 11 Apr 2014 NJ) the b.i.d.). nepicastat the respectively, in of Example inhibition DA/NE day concentration. animal. and levels days changes and state significant significant [0256] [0255] [0260] [0259] [0258] [0257] 10 13 plasma the mg/kg/day post drug to basis 7 DA/NE 60 7-9 at of following and yield Nepicastat Administration When The Thirty-two Chronic approximately Dog mg/kg/day). dose. in ratio. the on and Dogs 16 DA of for were day decreases plasma decreases peak all doses numbers N-acetyl the ratio whereas day (7.5 determining compared (5 receiving 6, The days. administered enzyme These (14.5 4 mg/kg 12 respectively. decreases were of treatment was NE male fold) changes post-dose. in 5, in metabolite the 1-16 The The 4, were days) of weighed plasma changes 15, significantly plasma and beagle b.i.d.), dopamine 8 peak to 0 nepicastat peak were and initial and mg/kg/day whether significantly in DA/NE groups the in as administration increases 6 NE plasma plasma dogs, 30 free NE 30 levels The assigned of days and reflect placebo dog mg/kg (2.1 β mg/kg nepicastat (n=8 and base -hydroxylase. ratio (2 the effects put different weighing post-dose, were weight received NE mg/kg, EPI 77 fold) in inhibition EPI steady different per as equivalents. into per (13.6 group, (15 plasma and observed were dose on of and and group): capsule were capsules was mg/kg from bid) plasma fold). 10-12 nepicastat EPI empty respectively. state group EPI nepicastat significant significantly from DA of used detected produced placebo were nepicastat (given (1.91 b.i.d.), at kg, period the and NE, A capsules (size placebo 2 to were and days observed DA/NE sympatho-adrenal (2 fold) DA determine on b.i.d. (2 increases or on 13 significant was dog mg/kg, The different randomly any post-dose. at mg/kg, 60 on and - all (placebo). and to 0 Torpac; numbers changes a ratio mg/kg/day days of mg/kg/day days yield at EPI significant period the bid, in the day b were from 4-9 plasma plasma post-dose. attained assigned days. doses i East No dose in po) 17-32 6 and All d) of observed plasma placebo system and (30 significant (placebo), produced produced Hanover, increases changing of levels days doses for DA as steady day mg/kg to 10, dose each The DA one and 11 via on 30 of of 8, at 2014202047 11 Apr 2014 plasma plasma AM were minutes. perchloric was vein, 4°C). homogenizing ice. Homogenates given dog animals group (pH supernatants 7.4 at sample and divided catecholamines cortex anesthetized from surgery. administered centrifuged conducted 8 [0261] hr -70°C at was following stored given dose, 7.4 the The and shaved. 8-10 B. and 4°C) Ventricles compound taken Membranes into compound studied for weighed at jugular Approximately delivered pellets 3 using (~80 The cerebral hr acid, at 32°C). The use ml using at 2 combined. with as from were later. -70°C portions, the buffer Dosing terminal of 4°C in mg/kg, scheduled. by per [ vein pellets 3 were frozen and were pentobarbital dose H] levels. blood receptor the a cortex centrifuged HPLC to levels. Non-specific were day. Dogs Polytron for and CGP-12177 and for skin left the consisted additional washed iv). homogenized the The surgery were determination were 6 in determination The Two twice were stored stored ventricle necropsy were regions binding using hr second The combined The liquid Blood obtained P-10 dog after or at washed then by Na dosed in AM for of at left 3 at binding 500 electrochemical in 3 was overlying studies. 50 resuspension of ml was the (~40 tissue oral nitrogen room -70°C days samples rapidly tissue buffer -20°C dose ventricle, in which x mM blood as supernatants then 78 from of AM of immediately g by 50 administration mg/kg), scheduled before where was was for disrupter tissue plasma until harvest containing Tris-HCl, mM administered harvested, until dose, resuspension a were samples both were and 10 jugular administered renal defined the Tris-HCl, required. and minutes an compound given cephalic, analysis. immediately a detection. compound stored was were put on (setting additional final first artery, frozen were centrifugation 0.5 vein 50 weighed, days by of performed into i.v. centrifuged the compound mM and mM 3 at collected Saturation one and 5 for 10 saphenous in kidney, ml as in levels. AM 1-3. 10, mM tubes levels. the The -70°C Tris-HCl, liquid μΜ All determination EDTA dose scheduled blood frozen a capsule centrifugation put 2 dose, cephalic supernatants On Na2EDTA x tissue over isoproterenol. PM renal containing for into at of administration at nitrogen A until 15 day The sample experiments buffer in and 48,000 and 500 pentobarbital determination third with dose 2 0.5 2 second liquid on medulla, 4, samples dog ml or days jugular at analysis x mM prior the of the buffer (pH transmural saphenous 1, and was iced g was x stored was once nitrogen baseline heparin, 2, with g and bursts). days second EDTA stored for to 7.4 taken 4 veins 0.4M Total renal were were each then then (pH and the the Na for 20 on 16 of of in at 2014202047 11 Apr 2014 bound times using bound, by mg/kg/day the yield the was mg/kg Analysis rate. way nepicastat-treated mean Membranes individual observed 60 administration 30 60 calculated dose at each gamma after determined 85%, [ [0262] 3 H] each minutes. mg/kg/day and final 10 1711%, used analysis-of-variance CGP-12177 catecholamine for first respectively. doses for for a mg/kg/day Student-Neuman-Kuels Dogs in non-specific 60 = Brandel globulin dose administration. each each 3 to of in a t-tests converting for free). mg/kg/day nepicastat seconds. by of were determine particular Samples tissue were were the 1767% parameter. all treatment significantly 10, liquid of cell as ranging were assayed nepicastat three groups 30 10 30, orally carried Individual Dopamine the and measure bound harvester. Aquasol scintillation and significantly were mg/kg by total and and tissue run any doses standard. group. In plasma with (ANOVA) administered 180%, for from For 60 60 out the to filtered and differences ligand (p<0.01) 1944%, group. protein separately. within mg/kg/day mg/kg/day group. or the compare levels scintillation example, renal using saturation 0.016 and total Samples Tissue 273% plasma compound placebo counting. Receptor (p<0.01) concentrations over with Fisher's artery, were using the respectively, count Dunnetf decreased nM 0, catecholamine Additionally, and in 79 each three 0.1% context wee for should Pairwise factor nepicastat, 5, to binding (control) significantly isotherms the fluid tubes nepicastat 268%, densities increased 15, 2 4.5 Saturation levels dose LSD s washed times PEI Bio-Rad nM. test DOSE norepinephrine or was days of be were between level to pre-treated respectively. analyses 30 treated tests compared a to were the Samples comparable added dopamine were were one-way free a with levels administered and mg/kg the control was set (p<0.01) protein binding to level linear were performed dopamine/norepinephrine tissue groups. up ligand a room expressed, performed the to between completed were factored nepicastat were of for GF/B each the to ANOVA. binding vehicle performed orthogonal levels levels isotherms increased was compound Doses temperature to eight analyzed concentrations experiment-wise placebo. incubated A at vial the glass treated harvested nonparametric in doses level for per by were treated method concentrations of for capsules and compound 2 each 86%, fiber A mg were at as 10, by ways. of contrast present of radioactivity each and significantly paired doses at water protein, validation. comparing its Following 30 10, group tissue and filtermats 6 81% analyzed 32°C controls b.i.d. hr (total partner and 30 tissue. of using First, at three t-test error level ratio after one was and and and and for 10, 60 10 as of to - 2014202047 11 Apr 2014 Norepinephrine mg/kg/day mg/kg/day, mg/kg/day were between mg/kg/day mg/kg/day placebo increased obtained increased significantly dopamine/norepinephrine (p<0.01) suggest 69% levels significantly 30 significantly significantly 502% decreased significantly 85%, [0264] [0263] 10, 10, 30, 30, and following determined 58% and were In and Statistical and the animals. each 60 on the by (p<0.01) increased 174%, and nepicastat, nepicastat 60 nepicastat. nepicastat, data 60 208%, norepinephrine (p<0.01) (p<0.01) (p<0.01) Day (p<0.01) mg/kg/day (p<0.01) renal significantly 150% dose mg/kg/day mg/kg/day. 79%, 60 are levels to compared mg/kg/day. No 4 analysis by cortex, respectively, not level be and respectively and 632% elevated increased significant increased and compared 86%, were increased norepinephrine increased linear): linear, Dopamine 156%, and tissue nepicastat. nepicastat, ratio compared In 612% (p<0.01) indicated 66% was not to and the factored with by placebo. respectively, by by and significantly was to in 522% and renal significantly changes at following 852%, 411%, 86%, placebo, 555%, the levels 1653%, the these plasma to 85%, compared decreased significantly that levels medulla, levels left 80 placebo, while following at 279% were respectively were 636% 30 Following respectively, ventricle. doses. the 1440% administration and obtained of at (p>0.01) were dopamine mg/kg/day (p<0.01) observed the its concentration significantly norepinephrine 10 and marginally to 72% and exceptions partner and dopamine/norepinephrine (p<0.01) and significantly The placebo. 677%, The 607%, administration on affected following in following 1693%, levels reduced 60 dopamine/norepinephrine in and dopamine/norepinephrine Day dose. of the mg/kg/day. dopamine (0.05 respectively, (a increased (p<0.01) marginally marginally 60 of 5 at levels Dopamine respectively, cerebral significant (p<0.01) by was Therefore, mg/kg/day doses administration these nepicastat 63% of were levels dose-proportional increased of 2 531% Norepinephrine 10, at (0.05 doses. and decreased 10, compared decreased result significantly levels doses dose 30 at ratios at nepicastat. in 30 the after doses doses and plasma and would of 156%, points At of were were ratio ratio ratio The 10, by by 60 60 30 10 10 of of to 2014202047 11 Apr 2014 plasma. mM membrane. Waddell, octopamine MO). Example hydroxylase, Example as stored activities 50 0.5 column cell dopamine-3-hydroxylase. enzymes cyclooxygenase-I. cholesterol endopeptidase, [0267] [0266] [0265] [0270] [0269] [0268] 10 substrates. mg/kg/day mM M line KH Human NaCl. at Nepicastat An Bovine The On KH containing 2 SK-N-SH -25°C. PO4, 18 17 were studied, W.H. Plasma Kidney Kidney HPLC Day 2 results acyl by PO4, Methyl The secretory nepicastat pH The DBH NO reverse Ca pooled 5, transferase, (1982). pH column 6.5, assay was 2+ (day medulla: medulla: levels 25 demonstrated and As and method /calmodulin a, from 6.5, synthase, ml 0.1 shown evaluated D-mannopyranoside phase DBH 4): was was and therefore treated gel of 0.5 was M adrenal Anal. is nepicastat used used was concentrated NaCl. M was in HMG-CoA HPLC eluted based group NaCl Figure phosphodiesterase no 2 6X10 3 prepared for protein to to purified Biochem: glands it X X The difference measure obtain its with on chromatography is 10 30 at and in 81 4, activity <60 < the 0.5 a concentrated > was kinase all nepicastat 35 reductase, vehicle 30 from with and 60 highly the was ml/min. separation tissues ml DBH (p=0.077) (p<0.05) obtained (p=0.076) A equilibrated between inhibition the at an of removed II, time-resolved treated a selective 10% activity culture Amicon range examined had acetyl Fraction protein III, enzyme (Feilchenfeld, from and left methyl an group. data. by of with medium phospholipase using CoA IC50 quantitation ventricular Sigma stirred (> buffer enzymes kinase containing solution were a, 50 A of 1000-fold) assay tyramine synthetase, D-mannopyranoside lentil mM Chemicals > of exchange higher N.B., cell (non-selective) 10 the including was samples of KH of lectin-sepharose μΜ most using neuroblastoma Richter, and aliquoted 2 dopamine tyramine than A PO4, inhibitor at 2 acyl with (St. enzymatic , all ascorbate a from those tyrosine pH neutral YM30 Louis, the CoA- H. in 6.5, and and and the 50 12 of β- in in & 2014202047 11 Apr 2014 were tyramine were tyramine. performed resuspended used without was UV hydroxylase incubated KH2PO4 KH2PO4, MeOH, of The solution 0.1 0.5-2.0 The The column Cf dose acidic for μΜ [ [0272] [0271] 10% 14 C]tyramine 8 1 mM each detection used ml/min samples CUSO4, assay light response reaction to methanol. incubated added acid, Enzyme Purification was μΜ inhibitor, 2 tyramine, wash containing reaction) containing and for pH at load ml in The was using washed CuSC>4, 10 37°C in to integration activity (150 200 4 2.3. ascorbate at of curve as was and H mM column total the mM 280 at Assay performed 2 50 ml substrate The O, corrected a was for μΐ) and A 37°C elute of reaction LiChroCART initiated 25 30% to with 0.1 nm. 1 mM counts and volume ascorbic utilizating vacuum -heptanesulfonic 30 were remaining obtain 4 [ mixed by mM was (two 14 mg/ml and mM minutes. stored for the C]-Tyramine. acetonitrile. 6 PC- KH2PO4, Radioactive ml and loaded at using of added data EDTA 30-40 columns mixture containing 1000 column the ascorbate. by with manifold of pH acid, each catalase at a high-pressure 50 IC50 analysis. the percent internal -20°C. C18 125-4 to to 5.2 software The tyramine pH minutes. and reaction mM O.f initiate a and values. by combined addition The acid, and column reaction Gilson 82 (6,500 Method. 2.3, (Speed 100 240 RP-18 [ In mg/ml KH2PO4, vacuum. 14 standards activity then The C]Tyramine eluate 37°C a (Thermo mM 30% 12 the and was μΜ typical u, The autosampler liquid mM HPLC to Mate column of was a reaction into catalase Boeringer NaAc, inhibitor -150,000 3-hydroxytyramine was in acetonitrile, Enzymatic substrate separate pH was reactions ascorbate/catalase After tetrabutylammonium and stopped assay, 0.125 Separations one) 30, chromatography. run 2.3 lyophilized and calculated hydrochloride pH fitted (final from and and was in that loading Mannheim, and the M 0.5-1.0 5.2, cpm. isocratic mixture by were the activity eluted to various and carried NaAc, Applied was volume product. analyzed the 10 reaction a products, to Bovine quenched nonlinear based then of mM milli-units addition washed with containing remove elution (internal was out [ mixture was 10 200 concentrations 14 Indianapolis, Separations) fumaric 4 C]tyramine, by 122: buffer phosphate, The 2 on at mM DBH ml assayed Fremont, purified ml μΐ). with the of HPLC with 4 acetonitrile, by the of of assay 124-128.). of parameter standard). 100 fumarate, and flow acid, catalase, (0.18 Samples at the enzyme 2 50 50 control 10 37°C. ml using using ml by CA) mM mM mM stop IN), was was was rate and 0.5 the ng of of of a 2014202047 11 Apr 2014 to more more by that mM bovine value. background, without KH 25 4.7 inhibition human identical at counted scintillation 50 column Compound dependent calculated 8 [0273] [0277] [0276] [0275] [0274] nM) be the mM mM 3-fold 2 ± PO the tyramine, competitive potent 0.4 potent caused following Nepicastat Nepicastat DBH. A The The 4 DBH EDTA, (two S , for KH2PO4, the in nM Km of enantiomer pH against inhibition from B IC50 IC50 14 each than enzyme bovine fluid combined were converted C than was (the a of 2.3 4 radioactivity. major 50 the values values mM 0.6 SKF-102698. tyramine with reaction bovine calculated was pH concentrations R (16 obtained buffer mM SKF-102698 DBH slopes mM of enantiomer). was ascorbate (nepicastat) shift 2.3. tyramine, a ml) DBH caclucated for into KH2PO4, to potent was DBH used by twice, which in and SKF-102698, concentrations: activity was Elution using one) by nepicastat activity Km, determined inhibitor at to Y nepicastat linear and with added by of that pH was followed pH was intercepts the obtain as for The in 0, into 2-fold) 8-fold on would 2.3. a 5.2 regression. was nepicastat, HPLC carried 1, more nmol/min, Ki 83 inhibition appears to both of 2, scintillation nepicastat and the from of (the pre-washed Entire the both 4, by against against be 0.5, potent and 4.7 bovine assay background. 37°C. 8 out S predicted the scintillation to nM 2 human ± enantiomer) linear mixture 1, Compound of be ml and as 0.4 Lineweaver-Burk than at were and the 2, bovine bovine and All competitive vials described the nM. of with 3, plotted regression and human Compound the three human for used the was 4 substrate was bovine MeOH The vials DBH enzyme, mM. R a B is same compounds to competitive loaded enantiomer (1/V 2-3 enzyme. and above. carried enzyme. data study with and by was DBH. The fold and concentrations B SKF-102698 buffer. plot. vs nepicastat to and were against tyramine. the inhibition used out equilibrated more 14 A a 1/S). Nepicastat C It caused C18 inhibitor. 9-fold (Compound Nepicastat samples blank corrected with was to counts potent human ReadySafe light obtain Km' appeared 8-9-fold kinetics A showed 1 a control against of ml dose Ki were were with than load was was The and 0.1 for (1 Ki of of B 2014202047 11 Apr 2014 parameter this proposed reconstituted vehicle benzyl of Example of of instead individual of Example equation. standard dosing, gavage administered aqueous formulations affinity [0279] [0278] [0284] [0283] [0282] [0281] [0280] the toxicity Ki) approximately report). study. of alcohol, Nepicastat A At Dose Competition The Clinical to of mice with at radioligand vehicle 20 19 clinical each competing all single the 1.5 logistic and body affinity Food nepicastat in other single were hours nepicastat selection time death mouse and each observations weights oral and route equation. and had receptors of 10 prepared polysorbate as oral of filtration occurred binding treatment ligands dosing, using nepicastat dose specified water minutes nepicastat moderate of powder, was doses formulations recorded administration. a of examined were Hill by were based were rodent a data binding at of group vehicle 60-mg/ml 80. in each 1000 followed diluting formulations 250, affinity coefficients was withheld the before recorded calculated were on intubator. were methods and protocol. was for retained 1000, determined 84 or an analyzed the nepicastat for dosing 2500 by Dose evaluated relatively clinical nepicastat from acute before or shaking. 60-mg/ml and alphai from were contained The mg/kg. 2500 potency This volumes the (body IC50 in toxicity by IC50 observations dosing. oral used. formulation mice in low receptors deviation mg/kg the formulation were iterative groups The weight route formulation values for (pKi were bindings hydroxypropylmethylcellulose, 2.5 study of 6- obtained Beginning the was to < of did nepicastat. calculated and (pKi data using 6.2). 3.5 was curve up duration and selected not in assays was 20-mg/ml hours to are of prepared with directly. which the affect protocol-specified 3 fitting 6.9 60 administered not over on Cheng-Prusoff outlined before vehicle. because of Clinical minutes - the tabulated the mice Nepicastat 6.7). an use. by to pKi integrity basis interval dosing, mixing a using it signs were (-log after four The The The is by of in a 2014202047 11 Apr 2014 were behavioral by between present. test observation # Example incidental 30, euthanatized died cages, Charles central Changes and each soft/continuous abnormal Clinical/behavioral changes compared [0286] [0285] [0289] [0288] [0287] 100-mg/kg RXXCM 12" 100, data nepicastat initiated in after and high. nervous Adult Lower Mice The The Rivers were or the were 0900 21 in Abnormal with and food tests. gait 300 group, dosing period, locomotor purpose 16) these 30- and in Surrounding male present. the 60 obtained. Hrs. body the mg/kg groups system. Labs. vocalization; produced and which and removed One and minutes mice behaviors but vehicle-control Sprague all and and water changes social temperatures of mouse posture, 100-mg/kg of surviving not of activity Rats were Rectal this consisted 2100 they nepicastat 6 No were the after from changes grouping the males were Dawley study removed these in in may treatment-related Hrs. 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One 0 mice mice metal considered behavioral incidental. end study. reactivity. inactivity, 18" (vehicle), lights behavior 102698 (model groups mouse in of group in x were were from tests wire and 18" the the the on 2014202047 11 Apr 2014 time numbering minute treatment next. response to mg/kg), water/50% treatment treatment noise piezoelectric in Instruments, Digiscan of hours. individually on attenuating a another startles calculated apparatus, after [0290] [0292] [0291] white this be a a scale a of Plexiglas® burst the five fifteen Acoustic Next, Each Prior for noise will injection and 1 Analyzer between groups) of groups groups for four most msec) and 32 minute enclosure. polyethylene every be their in 0 San to day the accelerometer generator minutes per each to hours. after a disregarded). drug rats appropriate 4095. cylinder startle Diego, after vehicle, of was was motor 20 box. rats, were such acclimation (model rat 40 a were seconds administration, injected the Acoustic presented this by but 5 running The that pg/kg. reactivity as CA) activity minute glycol), taking dH rat # (10 placed a testing transforms follows: number DCM-8). high 2 each for that for automated O. 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Treatment no less computed report. statistical vehicle-treated multiple measured. were the included and of within pairwise Pairwise time were Dunnetf were rest (or horizontal nepicastat ranked by trials computed for at Trials 87 were three called time each treatments. plotted comparisons. analysis. more comparisons within Date 2 the groups significant s terms comparisons 181-240 and data t-test. The startled measured. at separate 200 1-60 were sample), was activity rest controls, 2.5 for against 2 each (nonparametric mean for were milliseconds hours time were a hours, The each to compared Therefore, rat blocking the differences time within models time=4. (number maximum compared trial a over for than startle the treatment set (all day Each two-way significantly and treated clonidine-treated number to the time the p the to for factor the responses for The parameter time=l, technique) < immediately of at entire vehicle-treated vehicle each were rat to (TREAT) 0.05). startle and photobeams any groups each analysis mean and for the was 200 more time average of of trials treatment. using each responses baseline vehicle-treated were maximum the Note was interest to to tested milliseconds, examined at of movements the succeeding test group objectives treatment. 61-120 Dunnetf each analyzed analyzed variance broken), voltages controls that vehicle for (date), startle to were trial The had and the the the to in s 2014202047 11 Apr 2014 movements maximum responses times than test not vehicle-treated treated had <0.10). or first examined (both 3 0.0352). at average first significant significant [0298] [0297] [0303] [0302] [0301] [0300] [0299] 102698 for any rest significant revealed significantly 2 2 the the 1 hours time hours p time. Nepicastat Nepicastat with In When In Clonidine SKF-102698 and startle behaved clonidine < The vehicle in startle than general, or at for differences 0.05). 2 than 3, and the and that average maximum times for compared either (p>0.05) 10, controls response vehicle rest remained stayed response, less the maximum nepicastat group similarly had had administered 30 group the 3 time. Note (100 startle startle vehicle-treated or horizontal and no horizontal statistically at at between low at startle 100 at were (p<0.05). to mg/kg) time any elevated significant that 4 any the time response the response for to 30 startle mg/kg for time marginally response time overall mg/kg vehicle the the 1 vehicle-treated at both 2 activities SKF-102698 was (p activity 3, was response, for examined last of significantly SKF-102698-treated for Baseline < measurement. in controls 10, effects responses not nepicastat had the treatment 0.01) 2 88 marginally rats (PEG) either at hours. 30, significantly statistically last and and time a at significantly and on and or in 2 response. at any average significantly number and for 2 controls, the hours. 100 the (p< were Similarly, at effects 1.5 at and lower time significant time horizontal both vehicle time locomotor mg/kg 0.05), and the significantly not lower startle when of 2 for maximum The the 2 startle average higher group 4 significantly movements for the and does for less hours nepicastat were than compared SKF-102698-treated overall activity, average response (p=0.0703), rest activity average marginally movements responses average had not the startle detected (both different time and appear water treatment significantly startle no. different decreased and in to startle increased was response p startle average vehicle. rats. of and group. < significant vehicle at from at to at movements response statistically 0.05). all only 2.5 effect Dunnett's any effect response from Animals vehicle for at for times. startle group hours SKF- more were time time (p No the the the the for (p at = 2014202047 11 Apr 2014 noise times, temperature running. body between ml/kg randomly mean morning Example of hours. and apparatus and Clonidine administration groups groups so day, dH for for SKF-102698, [0304] [0307] [0306] [0305] 12 acoustic 2 groups that ten O acoustic 90 thirteen the core burst or for dosing and could The The The day. Since seconds Previous were the dosing rats, cyclodextrin 22 daily placed the and successfully temperature, of behaved startle. Motor on following effects treatment animals reading The not days eight but startle it volume. balanced 50 last after of average dose was will schedule work be in a time mg/kg, clonidine prior twenty rats high of similarly activity one balanced a impossible be reactivity of taken were chronic (SKF-102698's behavioral five has in groups lowered The each used) motor once nepicastat, degree across of to between was bid; shown minute startle the on tested rats the and tests consisted to a across for staggered dosing activity, eight were clonidine, dosing minute of vehicle since both the to first were d-amphetamine tests 20 that response consistency morning test acclimation were in and eight the treatment minutes. maximum balanced dosing of vehicle). there dosed an this were all of day (a acoustic during 89 startle nepicastat SKF-102698, such conducted one variable enclosed 20 96 motor matching ten and was administered is in day rats pg/kg, that rat within later The groups chambers. period significant the across and (about Rats evening startle on calculated from activity (8 the only inter-trial in morning startle times. average room dosing treatment procedure were bid: rats immediately rats rats (nepicastat, they days. and reactivity, 3 each 8 dosing hours rats during from were responses d-amphetamine, chambers, dosed with were variability 20 day were for startle and of interval were Furthermore, groups, minutes one the to examined. ten). and will each a in placed and by presented and be 5, white response eight day the run after oral were 35 be the after 15 ranging rat. however, Motor pre-pulse in n=12) evening to prior every between minutes or inside gavage treatment most noise startle the the measured chronic 50 The Between 2 with during all activity on to next. mg/kg, between day. mg/kg, appropriate body the every inhibition rats treatment treatment with generator the the 6 response after all dosing; and groups earlier startle These and same three daily were tests a core 8dB bid; bid; day the 30 10 10 a 2014202047 11 Apr 2014 the used potential were pulse the A these time by burst this operating of in immediately obtained dopamine chambers decibel 0 subject's attenuating (San agonist), sensitive dopamine days and average [0308] [0309] 15 to the piezoelectric a minutes analyzing colon eight assay. three 4095) as of Diego one, Plexiglas of run speakers inhibition mean. Acoustic Rat the meter noise 1 of movement immediately and sensitive startle of assay and properly. prior for msec) five by levels and positive body enclosure. prior Instruments, the each preceded d-amphetamine any Additionally, one with half was a produced and to cylinder PC norepinephrine to three detection were tests startle accelerometer core to rat. (Dietze decrease each hour. used hour assay a the ten. control computer into the Motor rapid temperatures prior Each reading were presented by Acoustic test effects reactivity to after daily a San Body to a (10 A a voltage and apparatuses onset), calibrate 68 in for a activity session to rat's the non run diagnostic the Diego, SR-Lab (a equipped cm administration startle dB the this was Kuschinsky, core of effects dopamine noise attached body startling levels, via morning concurrently and level ten diameter) (a which DBH assay. to were has the temperature calculated. CA) a reactivity series calibrating day core pre-pulse assure bursts speaker to of with 90 of program speakers been which was inhibitors obtained stimulus) test ± below daily chronic DBH releaser) background temperature of 2% SR-Lab which of (a 1994) that shown then stations. muscle immediately which mounted broad dose clonidine makes inhibition of inhibitors instrument in has was Body the the rectified by dosing the were were which each in-vivo. was to of software photo been inserting band occurs plexiglas run contractions noise mean. be was The this nepicastat, core 30 of used both housed schedule and (sensorimotor on sensitive in-vivo. shown makes noise and beams the measured rats was cm alter behavioral throughout when temperature each Both d-amphetamine, and measured as the Startle digitized eight above were cylinder used the with in the to interface this and rectal and (to elicited a of to clonidine D-amphetamine positive a be test startling placed motor to three reactivity a the obtain the changes light ventilated SKF-102698, behavioral rise sensitive calibrate test in gating all (on probe station transduced readings motor animal. by eight times test assembly. sensors time a activity individually controls a a stimulus (an scale an on 2 baseline), measured in potential sessions. and to cm SR-Lab and activity to and each intense central sound dosing alpha test ± Also, were were from both pre into was and test fall the 1% the for of A is a 2 2014202047 11 Apr 2014 noise trial noise minute pulse Acoustic photobeams motor reactivity these the bursts injected. in in consisted amphetamine (about calculated subjects avenge avenge are Clonidine shown after [0313] [0312] [0311] [0310] 102698, pseudo-random mesolimbic assessed. loud interval the behavioral burst paired bursts activity and 4 to Each The Three The of of acclimation noise startle hours and stimulus and startle these be of At 40 by startle and (118 schedule subject's that (and rats trial ranging placing sensitive temperature readings 3.5 pre-pulse subtracting on bursts is 10 and dopamine elicited three d-amphetamine the dB), reactivity test assessed. spontaneous reactions). (pre-pulse dosing hours, startle minutes order. was 40 period subject by a readings of reaction between and each minutes to (one potential inhibition by presented. daily 100 reaction the day the changes Pre-pulse a readings levels. the was At rat the relatively broke after per inhibition msec core The mean locomotion ten). behavioral was was 4 30 individually rats 118 after millisecond) calculated served hr. sensitive measured) three of and a body during (pre-pulse Furthermore, then in were measured startle dB 40 were inhibition Each The acoustic supplemental the dopamine quite 90 trial minutes, 91 different calculated. acoustic as temperature taken morning the tests startle was presented assay seconds response startle the by noise described into on testing inhibition starting startle during obtained has was from determining positive to average startle and acoustic reactivity types burst. a burst reaction daily been was the as SR-Lab administration elicited tests. session. each with is norepinephrine each immediately above) follows. of effects reactivity trial). (77 by used) measured. injection shown Pre-pulse once control subject noise startle one trial calculating was and dB), by the test from for These of of a to the bursts pre-pulse for At calculated the mean minute reactivity station DBH per of for and three an be inhibition after the 77 t=0, of the At nepicastat, quite trials hour time levels sensitive the pre-pulse the consisted dB response 118 inhibitors clonidine 3 different 40 DBH (a each and hr. burst acoustic total of inhibition pulse were (60 variable msec has by dB which testing. values 35 after noise inhibitor to total taking number alone preceding and also inhibition presented of - for minutes, types window changes in and 118 makes a a startle burst. inter noise SKF- vivo. were been each tests loud trial five The the dB of of d- is 2014202047 11 Apr 2014 performed treatment treatment using within using trials value] by trial multiple hour. over inhibition Differences (nonparametric each - and adjustment adjustment displaying and startle (RATIO) groups. [0315] [0314] [0318] [0317] [0316] pre-pulse the then these TIME (TRIALN). time 1-15 the Analysis treatment, Each 118 response An The These The Spontaneous comparisons 118 If dividing were error plots trial by effects for the inhibition overall correspond the db for on was was mean average was db time values each time alone mean described each overall the term computed are trial technique) of applied made. elicited used time this Pre-pulse average block by one-way Variance. comparison attached change treatment value). effects trial startle motor percent were for trial is value time to significant to and then animals to above) (every If for by value] time adjust for plotted voltage treatment was response activity the the by ANOVA from to were were inhibition made. prepulse each the each are The the the of 1, specific overall performed from 15 for nested attached 77 interest. baseline difference 16-30 rat, against 118 studied. not 118 report. treatment (AVGMEAN) model min) was multiple and by dB with the inhibition i.e. db db values 92 significant treatment within time pairwise time measured was pulse trial the 118 trial ([118 alone also. included a Note to for The is main (TREAT) percent 2, interaction. comparisons. dB value). test analyzed were number not treatment. each trial - dB and 31-45 that method alone comparisons. 118 and effects effect for detected, for (p-value> trial terms calculated the for the animal. prepulse the mean time dB each for and trial separately. each mean value y-axis for were of for Treatment difference each Overall pulse trial 3, and treatment animal Bonferroni's percent If rat, Fisher's treatment, 0.05) average - inhibition and by Subsequent on the nonsignificant, pre-pulse treatment then type Further, i.e. paired the subtracting treatment 46-60 overall every Kruskal-Wallis ([118 then prepulse effects between dividing (TRIALT) Least was plots startle trial animals were adjustment if inhibition time and dB a 15 performed treatment t-tests varies. the were effects Bonferroni Significant of min trial (pre-pulse trial this inhibition the treatment analyzed 4. then specific animals at nested tested value mean value for Plots type, were each trial The and test the for or at 1 2014202047 11 Apr 2014 time. treatment pairwise time the vehicle performed of inhibition. analysis. adjustment also Treatment statistically and significantly examined. 50 significantly core cyclodextrin significant significant SKF [0319] [0323] [0322] [0321] [0320] mg/kg treatment, first the examined. applied temperature group None The There The The 45 control treatment cyclodextrin b.i.d. A by to effect effects difference minutes Pairwise The positive was change d-amphetamine repeated significant group different different test to had of time was time group overall the made on the at the on were marginally (p effect no from (i.e. and body controls effects comparisons all treatment treatment measures day between treatment had = from from group for overall time 0.0782). samples treatment not for times pre-dose core was significantly one the the the were group effect significant (d-amphetamine cyclodextrin (p temperature not the effect vehicle of vehicle individual effects two-way significant higher examined. groups 1 revealed = the in not - two by significant was had 3), 0.0001). body at time chronic controls lower vehicle significant within but controls statistically 93 each percent significantly for produced ANOVA weights comparisons that versus not at treatment interaction. any day. locomotor The any and (p-value time. significant dosing, none controls at The at comparisons prepulse time. clonidine) any was dH any clonidine was (p-value any significant of treatment 2 higher time If effect O time the but used > calculated (dH within activity significant both One-way 0.05), (p after nepicastat-treated examined. inhibition no 2 examined. significantly to O > locomotor for = group, of the for 45 other and test 0.05) by time than 0.0283), then nepicastat interest. minutes. the for overall time ANOVAS for SKF's change its and compound the however, each SKF-102698 then Also, compared the The vehicle activity interaction increasing adjustment but averaging vehicle) overall treatment animal However, a SKF- there in at groups Bonferroni no were control any pre-pulse was than had 102698 was to others. for effects at group body were time over then was was any and any not the the the the no at 2014202047 11 Apr 2014 vehicle prepulse body pre-dose b.i.d.) b.i.d.) than nepicastat mg/kg of startle significantly different each from (50 significantly amphetamine controls groups cyclodextrin, for cyclodextrin for [0325] [0324] [0328] [0327] [0326] 102698 interest SKF-102698 mg/kg the the day, pre-dose weight group. reactivity. group b.i.d. During at Only The There The during 50 cyclodextrin than had from inhibition comparisons the the 50 (all b.i.d.) d-amphetamine SKF-102698 mg/kg versus versus different the different pre-dose than mg/kg had was No a the SKF-vehicle and group p times their baseline times significantly = SKF- 50 other group vehicle significantly The no 0.0001). the also dH b.i.d. SKF-102698 than b.i.d. own mg/kg 3 at group 1 overall from from baseline. 102698 overall SKF-102698 2 and significant O of and treatment showed than had vehicle (50 the controls group. (all group 4. controls dH b.i.d. amphetamine the group The 2, at mg/kg or its vehicle significantly greater p time 2 Neither treatment all the O higher vehicle < pairwise a treatment vehicle had 50 versus at at 0.05), times. had (p<0.0 differences significantly days clonidine effect any had any group b.i.d.) mg/kg increase control significantly a d-amphetamine startle controls 94 time. significantly but 4-10. time. a group cyclodextrin 1). control During significant was significantly by versus at group lower b.i.d. no When group response treatment time were and statistically produced others. None from smaller The at times (SKF-vehicle). (p had startle any were dH interaction detected. analyzed lower had = clonidine differences smaller pre-dose of 2 (p significantly nor 0.0047), O, time Treatment than 1 increase greater days just the not = a and response significant SKF- startle clonidine significant 0.0007) examined. nepicastat the within change significantly significantly 2-10, 3, was group, in increase but the 102698 (p<0.01) in SKF- effects When response body statistically lower body no except and nepicastat as each in for versus 102698( change however, others. body dose compared The was for from weight analyzed were weight day, all startle in higher different days nepicastat compared SKF-102698 body comparisons groups significantly weight pre-dose dH the in significant significant (50 The 50 response than 3 between was acoustic 2 percent vehicle O weight to within mg/kg and mg/kg SKF- were from from and not the the 50 to in 6. 2014202047 11 Apr 2014 respect the time nepicastat-treated nepicastat tended non-lesioned taste. marshmallows nepicastat. prior weight b.i.d.) mg/kg Importantly, MPTP individually of Inc, Example and drug (ii) approved available 0.95 [0329] [0332] [0331] [0330] 2 SKF-102698 oral the (Natick, ml examined. mg/ml to administration group Oral b.i.d.) Monkeys There Six to l-methyl-4-phenyl-l,2,3,6-tetrahydropyridine (kg) vehicle 3 commencing of to syringe, 23 ad by months changes MPTP at squirrel spit syringe Three of libitum. (free at there housed the MA). and group was any was days control each Interestingly, Institutional out (50 were three and at base) no was groups different in prior), time tested For injection monkeys, 2 7-9. had due and animal. behavioral mg/kg body All the overall mg/ml (ii) no groups maintained lesioned administration, and significantly examined. procedures to allowed were difference solution were oral in weight. approaches b.i.d.) failure was resulting significant Animal of 3 For there on three gavage. non-lesioned significantly studies. nepicastat monkeys used any injected a example, group on When was at of minimum non-lesioned used in day. Care Pairwise lower to in a animals the 95 changes MPTP were a treatment Insertion 2.0 13h/l and study in was analyzed significant subcutaneously (0.5, and highest body this a mg/kg monkeys examined different the 950 also was of lh Use to comparisons in the 2, study nepicastat one weights light-dark and effect ingest body of and gram suspended not within final Committee drug optimal (p<0.05) (MPTP) nepicastat month followed three and from an 5 including weight per on animal mg/kg) treats in acceptable each than concentration. proved body (50 injection. lesioned the cycle, a route revealed to in overall was volume (IACUC). day, between the probably NIH mg/kg received solution acclimate water vehicle into (i) weight to purchased nepicastat with of the insertion guidelines route difference the be (received at that administration b.i.d.) (ml) SKF-102698 the food controls a for a (5 due mouth an Animals to poor since concentration Oral none SKF-vehicle mg/ml) equal any injection group the into to from (50 and and 2 between route of animals adverse dose gavage colony of at mg/kg mg/kg treats, to water three were were RBI, with into any (50 the the of of of of 2014202047 11 Apr 2014 was pm), received mg/kg base-line using period recording was mg/kg mg/kg/day treatment monkey mg/kg) MPTP MPTP Parkinsonian Both at administration lesioning. consecutive spontaneous drug Group administration [0333] [0336] [0335] [0334] the not administered nepicastat the for a 5.0 twice of A lesioning 3 (free-base) Animals Prior Twenty Six parkinsonian and replaced. receiving nepicastat months activity 5 activity mg/kg (6 2 sessions groups, Locomotor days (2 squirrel was weeks. days. animals) daily). motor model. to mg/kg that was at four well were dose. behavioral with prior), monitoring of via lesioning, 5.0 1 with via were resolved Normal at monkeys, each mg/kg/day carried In The activity squirrel clinical accepted. twice oral received mg/kg a a activity lesioned The subcutaneous Drug Group were concentration two-day 6 60 behavior animal. syringe animals twenty-four minutes assessment upon daily); animals out (IRAM) was rating had used animals monkeys, using B, was three placebo by (0.5 in washout one well light one at the to of monitored per scale and 3 in mg/kg non-lesioned the an did injection and study animal animals day of tolerated beige MPTP-lesioned administration length were was 96 group. (water) animals Group fourteen infrared 0.5, 0.5, not (CRS) clinical withdrawal between safety twice carried colored died 2.0 typically 2.0, subjected and by was to D at treatment; once The were and IRAM rating females or achieve daily); the acutely activity were and (6 and also the out 5.0 loose per animals) 5.0 groups two of of tolerability randomly score three monkeys 2 different carried assessments assessed mg/kg drug. in MPTP to Group mg/kg day to following lower and a Group monitor stools 60-minute 4 parkinsonian quantitative lesioned greater consisted (12 weeks received ten out twice at C doses doses. assigned dose by noon on at B of (6 males a for MPTP-lesioning, (IRAM) the (5 established the 1 nepicastat. concentration after than animals) daily, (received and levels. sessions to to 10 animals) of One final 10 highest 3 were 1 the sessions 3 assessment state. to the as pm) mg/kg/day clinical (10 non-lesioned on one two oral last cage. Nepicastat received following: used for the am for the 2 Animals received A dose days MPTP- of gavage mg/kg over of 3 3 raters and mean CRS. post- four in to to and All 2.0 of of (5 (5 4 2 5 5 a a 2014202047 11 Apr 2014 parkinsonian behavioral total period monitored. treatment minimum treatment. obtained hour immediately on study. sufficient days. sessions lesioning following 5 am different CRS. (at administered Testing for conducted [0337] [0340] [0339] [0338] days the each 10 and monitoring clinical Clinical of am Clinical Nepicastat of Animals In For last This 2 was (IRAMS). animal. 3 locomotor within drug drug treatment degree some 2 pm groups. in and There to assessment 4 The day statistical the state final following at 5 days carried rating by rating administration 2 treatment. days. behavior a cases, were washout average morning, was pm) three sessions. of oral concentration The and or post-MPTP of activity IRAM lesioning Activity score groups. was a tested for out water pre-MPTP-lesioning treatment. used gavage. animals minimum analysis, (IRAM weeks drug locomotor (CAS) after carried 7 60 Raters 4 as greater consecutive The was assessments for (as to to administration monitoring at a to L-Dopa required of Behavior pretreatment of and IRAM response evaluation 2 90 was determined post-MPTP f2 out display placebo) 2 locomotor (one pm Raters commencing either activity minutes day than CRS), weeks 60 assessed on to dosing. 97 washout additional to assessment days. the 2.5, parkinsonian were 3. to three (one was was 90 was was baseline within was after by after L-Dopa last value at activity (pre-treatment) 5, minutes All rated reported by to individuals) preformed averaging Behavior Drug calculated 2 or period administered 5 the the used three one the animals pm days doses for 7.5 three by was and were efficacy 10 was statistical following and to on mg/kg last symptoms, between to of IRAM individuals) as am of determined the weeks was three three drug administered the pre- “ establish for received clinical MPTP preformed MPTP dose movements/fO were efficacy behavioral by determined study. for last and and treatment. to 90 individuals each analysis. the of of oral blinded five defined 12 (2 minutes 2 post-MPTP-lesioning CRS. dose. nepicastat starting rating were 10 by a a treatment to gavage mg/kg) of f-hour days The am final baseline twice for averaging 5 drug assessment to by blinded The as morning days minutes rating L-Dopa scores immediately the post-MPTP- the following 90 post-MPTP twice monitoring IRAM an to daily nepicastat. on dose. CRS different obtain efficacy minutes of average clinical the over ten to ” at daily were dose drug . was was was and last the 10 A 1 a a a 2014202047 11 Apr 2014 minutes through placebo parkinsonian three performed parkinsonian) higher individual Post-MPTP, each compare concentration animal. experimental analysis This (one score derived due and graph greater [0342] [0341] [0346] [0345] [0344] [0343] 7.5 to to average 10 was weeks was score IRAM Eight In insufficient Statistical Statistical than Statistical three) mg/kg minutes D). to from of following The pre- the obtained raters since nepicastat then variance). three versus was of behavioral CRS parkinsonian the as conducted animal. corrected drug clinical Locomotor and L-Dopa commencing no from data a examined considered sum on analysis analysis analysis ratio was post-MPTP-lesioning IRAM difference for 90 concentration each the blocks at data rating minutes of and each used This of 1, clinical state. CRS. over these activity 4, drug post-MPTP over data consisted consisted comparing nepicastat features to of drug and for comparison score a the the was placebo for detect A faster 3 eight collection. level. scores statistical 10 to dose consecutive post-MPTP efficacy was nepicastat. determined corrected measurements, 5 mg/kg/day were of: of (1,4, animal. features. consecutive any clinical post-MPTP by monitored and 98 are descriptive A (1) was activity averaging rated analysis. trends. study experimental clinical 10 higher Only by comparisons multiple clinical from repeated scores. mg The using For in the by every for 60 days. /kg/day no lesioning each Wilcoxon graphical rating scores A statistics Further the each averaging final the minutes rating each (2) pre-MPTP-lesioned lower dosing using clinical 10 animal drug Kruskal-Wallis animal, Pairwise or of between minutes is post-MPTP group score statistical placebo) score (pre-treatment) analysis. and the at experimental sign (with sessions considered the rating and 1, a average was graphing of 4, was rank comparisons for total the single the the and animals was scores determined were a analysis considered same ratings test average (non-parametric minimum total CRS 10 not a animal CRS clinical drug the was mg/kg. collected to faster of dose) (groups performed score of for all 2.5, mean for was between CRS used at within scored 1 rating a raters of days. (less each each each to was The 5.0, less not 90 of of to A at 3 2014202047 11 Apr 2014 Normal Normal the when repeated the Accept Accept than Accept Accept and Group groups animals Group groups scores Group groups animals Group groups [0348] [0347] [0351] [0350] [0349] 1, 10.32, 4, degree average placebo 3. and needed. range D: C: Β: after A: IRAM There There There There due due due due Null Null Null Null animals animals showed showed measures). 10 of 4 10 1 to to to to Placebo CRS MPTP-lesioning. Hypothesis mg/kg. Hypothesis mg/kg/day Hypothesis. mg/kg/day treatment Hypothesis. 4.3 MPTP-lesioning was was was was mg/kg/day the the the the (activity (non-lesioned) (non-lesioned) substantial substantial to post-MPTP high no high no high no high no 16.1. Treatment. The Dunnett significant significant significant significant degree degree degree degree using monitoring) Treatment. Treatment. Clinical treatment. The same The All All increase increase ’ lesioning Friedman average s Normal in of of of of average animals animals analysis typically typically post difference difference difference difference each variability variability variability variability Rating Wilcoxon Wilcoxon and hoc Wilcoxon Wilcoxon in in squirrel CRS animals (pre-treatment) CRS showed ’ 99 showed was s the the have have Score CRS analysis analysis between between between between for of of of of performed signed clinical clinical for scores scores monkey. (clinical movements movements movements movements signed (CRS). signed group (non-lesioned) signed substantial substantial group for (non-parametric pre-lesioned pre-lesioned pre-lesioned pre-lesioned rank less rating less rating non-parametric C rank rank The to A rank for rating was than than test: was 2.5, nepicastat per per per per average increase test: increase scores scores test: test: 8.97, 3. 3. 5.0, 8.9, scale) W typically 10 10 10 10 W W and and and and W = minutes minutes minutes minutes and range range after after = = CRS analysis = in in 19, post-lesioned post-lesioned post-lesioned post-lesioned were data at 21, 17, 7.5 9, the the concentrations MPTP-lesioning. MPTP-lesioning. N have 4.8 6.5 for N N N was mg/kg used per per per per clinical clinical = = = = group to to of 6, 6, 6, 5, scores animal performed animal animal animal 15.4. 17.3. variance, to P P P p L-Dopa IRAM IRAM IRAM IRAM < > > < B assess rating rating 0.06 0.06 0.06 0.06 was less All All for for for for of 2014202047 11 Apr 2014 Normal by parkinsonian nepicastat treatment than mg/kg/day points post-MPTP monkey. Pre-MPTP-lesioned Placebo MPTP-lesioned improvement improvement animals CRS groups scores 7.5 analysis dose activity statistics, Group different from significant significant [0352] [0354] [0353] mg/kg IRAM placebo 8 results level. after to 10 A Overall There due B animals monitoring all had of concentrations comparison and 10 groups to of Both for effect improvement dose scored at between to MPTP-lesioning. (pre-treatment), There animals, out a showed 90 L-Dopa at treatment. symptoms over Dunnett were borderline the 4 the there 2.5 (non-lesioned) of non-human minutes level 4 in (1,4, and greater high the a was and mg/kg of parkinsonian 5 animals total no was base-line of a demonstrated (animals when ’ placebo mg/kg/dose. s and post-MPTP 10 difference of degree 10 no treatment test compared detectable with post-dosing. no significant possible than L-Dopa Based nepicastat mg/kg/day 10 mg/kg/day difference compared with primate scored post significant The mg/kg both 3. treatment typically of (pre-MPTP-lesioning) of symptoms on the hoc All groups variability CRS when average the between (pre-treatment) to a 5 no differences effect. a model (1, significant of exception No groups mg/kg analysis post-MPTP to 100 non-pairwise same of of concentrations Ten-minute greater score difference have nepicastat compared 4, to placebo. and groups nepicastat drug CRS 10 all compared pre-MPTP Using of (A and of of group scores L-DOPA, mg/kg/day) was than other PD. of 24. (nepicastat) through the effect for between 7.6 of in to state. lesioning, Group the and At intervals done, group 3 less RAM examined comparative concentrations the post-MPTP. and mg/kg Based to animals in when and 1 same showed placebo), and Friedman than mg/kg/day placebo D) the locomotor All and placebo C placebo in C results post-MPTP-lesioning had post-MPTP-lesioning compared was for pairwise treatment CRS. on L-Dopa the were 3. (pre-treatment groups a pre demonstrated comparison an the a a MPTP-lesioned (water nepicastat 8.02, analysis test for pairwise plotted showed significant average treatment and animals Post-MPTP-lesioned of activity 7.5 of when results, comparison, each nepicastat range to at treatment) mg/kg post animals the the for of animal. CRS a produced compared as were comparative between evaluation). 4.0 post-MPTP treatment), descriptive 4 significant significant and each 4 measured effect dose different ranging to showed squirrel and at slower within states. in 5 three 15.6. drug time The and and the the no 10 to in 2014202047 11 Apr 2014 used. nepicastat were number to plasma maintain Example on on and demonstrated and lesioned continuously loosely compounds, cannulated animal days. study drug analysis days for for [0355] [0358] [0357] [0356] drug day day analysis. detecting tolerability Group dose fasted with where One One A variability A Male, This taped 1, dosing. 12. level of patency state 1 24 pharmacokinetic then of second in animals a hour milliliter milliliter C with overnight Baseline respectively. study 2-day each recorded the to animals of spontaneous Nepicastat no (no a in 6 study. the significant prior nepicastat. hours squirrel all of significant PE50 effect of by pharmacokinetic also is washout each restrainer. groups of of the used. plasma comparing then to were using after Three blood blood demonstrated tubing different at cannula the monkey. was hypertensive anesthetized Animals study 7.5 with drug tested Modular the effect. between first This levels was MPTP-lesioned (drawn administered Heparinized mg/kg first for throughout the the was effect dose drug drawn study on This were were study recording exception same dose Instruments from carried each L-Dopa) each that, levels. to with rats 101 study than was from determined then establish on animal was the saline drug the (280-345g; a at of ether. out day carried a pairwise squirrel was concentrations the femoral placed three of experiment. of carried animals. non-pairwise to concentration. MI 7, (50 pre- Group femoral carried blood A baseline determine 2 and different out units on BioReport™ femoral in monkeys and vein Charles analysis, out finally samples B MAYO concomitantly However, pressure out vein post- sodium (no to The of and of artery study concurrently the drug each determine River effect 6 6 (#353, following treatment, which 1, at Blood restrainers hours collected hours plasma heparin/ml) software 4, and 6 and concentrations animal) 2.5 design Labs, hours at and 358 reduces after after mg/kg was femoral 5 with administration concentration the parameters mg/kg 10 with is the Kingston, and installed when after was and the the collected better mg/kg the steady-state was week of 374) the vein animal first first their collected L-Dopa) this efficacy L-Dopa a used for suited safety on small for were prior were were dose dose NY) first feet for 12 an of to of to 5 2014202047 11 Apr 2014 bolus base baseline ANOVA ANOVA period treatment, pressure pressure baseline period pressure nepicastat IBM nitrogen minutes recovery deionized effect date dissected euthanized administration (deionized dosed (iv)/nepicastat (iv)/vehicle [0360] [0359] [0362] [0361] 120, 15 min to 150, concentration personal in was intravenously and of determine All At Following Recorded a blood for following and and and later, and with out, by time. period. 180, volume water time, not or water, for the via compounds each (po); heart Fisher heart stored vehicle weighed, heart computer: significant. the 210, a pressure of (po). animals end decapitation. and main Once (vehicle) parameter nepicastat catecholamine parameters 10 vehicle of post-oral rate rate surgical ’ of Animals intravenous and s at rate of with their ml/kg). 1.0 was LSD 0.2 effect Once baseline -70°C. was were the and 240 and were ml/kg were were mean either interaction. mg/ml. (iv)/nepicastat to 10 strategy determined at time fixed experiment, heart min. or preparation, for animals a were analyzed were ml/kg. specified The dissolved Biochemical orally vehicle, followed free blood analyzed arterial SCH-23390 time. administration levels period. in rate. Nepicastat measured cortex, randomly base with 0.4 were SCH-23390 pressure dosed Recorded This by Pairwise including pressure by recorded time M by each 102 concentration Further on a (po); separately. each an taking perchloric left Bonferroni 0.2 stabilized analysis analysis (200 points the with analysis 15 or assigned animal and ml SCH-23390 ventricle animal of (MAP), comparisons parameters parameters min saline dopamine day analyses an pg/kg) was flush either SCH-23390 heart in average was are acid. prior the of of was (minimum The correction dissolved of was were of to heart performed variance or rate performed (apex), use. experiment. nepicastat were isotonic 1 vehicle to Tissues and four anesthetized change (iv)/vehicle were were mg/ml. of allowed administered were were intravenous rate each Nepicastat performed norepinephrine or treatment and in then when measured one (ANOVA) (HR), performed saline. established, were vehicle. (saline, on from saline both parameter Oral (10 mesenteric a measured hour), these the (po); minimum and then with for baseline mg/kg) dosing dosing intravenously (vehicle) at was 1 groups: overall at tissues the the each ml/kg). with frozen Following baseline following or halothane 15, over levels. animals dissolved change at post-iv SCH-23390 artery to volume time 30, or effects 5, for one treatment at a to establish in vehicle vehicle Fifteen 10, 60, 15 a Blood a liquid blood blood by were were from hour each time later as min free oral and and 90, for for an in a 2014202047 11 Apr 2014 Nepicastat between vs. Vehicle treatment the to vehicle heart Hydralazine MN) Example in This comparisons. (MBP), least cannulated aseptic anesthetized chronically antihypertensive duration decrease significant [0365] [0364] [0363] [0368] [0367] [0366] heart the Vehicle experiment, was 2 for rate abdominal weeks conditions, An Intravenous There Male Intravenous Systolic during rate (iv)/nepicastat heart treatment of measurement 25 (p<0.05) statistically group during with did decrease the (iv)/nepicastat implanted additional observed (10 with Crl:COBS(WI)BR before were rate the Comparisons experiment. not the blood mg/kg, effects by the the musculature, groups. pentobarbital 15 in an (HR), decrease no treatment catheter an administration (p<0.05) being rats significant post-oral min over mean incision with pressure significant (po) of analysis ANOVA p.o.), observed were arterial and post-iv (po) the subjected telemetry vs. arterial Pretreatment of the of nepicastat with course in was motor period were a the Vehicle (SBP), SCH-23390 randomly (p<0.05) was heart telemetry sodium rats with mean with blood period. differences made skin SCH-23390 pressure of made. to of implants performed activity of at nepicastat rate a diastolic (iv)/Vehicle arterial SCH-23390 this was pressure, (30 drug 103 120 main on 15 at (60 with Oral divided transmitter as mg/kg, 150 experiment midline. weeks (iv)/Vehicle closed. min administration. by in mg/kg, much (MA) SCH-23390 effect (TA11PA-C40, treatment resulted pressure and blood baseline administration 30 to heart and into p.o.), old (po), min 180 as produced compare were Each for The unit. ip) 240 pressure rate should observed were in 4 min compared post nepicastat with heart treatment (po) and and abdominal treatment min a monitored. rat did After and significant post Three SCH-23390 used. dose nepicastat vs. was its the be not rate Data a compared alone. (DBP), motor in the noted. dose. small Vehicle abdomen significantly baseline allowed or (100 to and which days vehicle Sciences, groups: transmitter aorta Twenty-four mean animals activity. mean decrease Both subsequent The mg/kg, (5± caused prior to (iv)/nepicastat(po) continued (iv)/Vehicle was to treated shaved. arterial means large vehicle Vehicle blood 1 nepicastat recover Inc., to that exposed, attenuate p.o.). mmHg) a The was (p<0.05) the significant variability rats St. pairwise received pressure animals. pressure of control. rat sutured start for Under (p.o.), for Paul, were (po), each was and and yet the the of in at 2014202047 11 Apr 2014 time time were were the hydralazine however, At tended performed pairwise was DBP, 2 rats 0.05. study. comparison, dosing. dosing approximately after day overall [0369] [0374] [0373] [0372] [0371] [0370] 10 at hr, 100 rat received hour 1 detected, and points expressed . then dosing MBP, and Data in on As to Nepicastat A After Throughout Hydralazine mg/kg, Oral treatment Significant their comparisons 10 Nepicastat 13. slowly computerized day the a by averaged on were measured ml/kg hydralazine HR, maximal administration a the on adjusting Similar effect interaction 1 a -17 7 MBP, the . as day series pre-dose day lower Similar and prepared effect means mmHg and at but compound the progressed, at hourly daily at 30 HR MA. decrease degrees 1 each were at less 10 of the study, 100 blood data and (p<0.01). was at transient ± each was and on treatment mg/kg one-way values in critical pronounced SEM. mg/kg 10 of day. Data and expressed collection 100 day water detected, none pressure MA induced used. of nepicastat mg/kg of time a standard antihypertensive mg/kg, 3 caused for on -24 peak A hypotensive Statistical value caused were (p<0.0 of ANOVA with of MBP point two-way each If these mmHg induced the as vehicle, a but hypotensive system an then bradycardic using analyzed 104 peak traces did an free at drug 1). errors rat a overall were parameters continued did bradycardic immediate 30 significance the not (p<0.0 were The base at ANOVA antihypertensive varying a was treatments effects not nepicastat of mg/kg Bonferroni each performed of consistently pairwise treatment MBP separately. effects Tween equivalents. induce collected used the effect responses 1) to time were in were (all degrees hypotensive mean remained with response to MBP was decrease were or showed 80. of difference a doses continuously point observed adjustment. using effect established, hydralazine. consistent -10 every affect main defined (SEM) were Each All was induced of response mmHg expressed would of low a tachycardia doses or Dunn and 5 observed effects consistent observed HR analysis effect -100 throughout a min. from as calculated. throughout hypotensive significant throughout reached on was ’ a respective were be collect of s b/mm p which for procedure. for day control -11 hereafter performed. level observed within was on effect throughout given 10 treatment 1. mmHg data 3 days the its the subsided done of sec. hours interaction All On the would groups 1 effect study. less on nadir orally on study. 3-7. hr are on values day These study. on MA. 22 If SBP, after after than The and day po) the no 26 on be In hr of of to in 2, 2014202047 11 Apr 2014 weight to pairwise with nepicastat Urinary receiving the nepicastat Dunn respect of Example of of Example (p<0.05). and symptomatic frequency dopamine after [0375] [0381] [0380] [0379] [0378] [0377] [0376] nepicastat. that dopamine the treatment mean the 24 ’ s rashes treated Nepicastat on Using Body Four In to One Shown procedure hour dopamine rash. comparisons 27 26 the patients 200 to day of administered activity Although supine patient of or norepinephrine. significant treatment orthostatic by weights changes the were 3, with mg in 8 increase day it patients Figure repeated-measures and with reduces levels of was plasma treated measure vehicle, accompanied interaction. treatment for nepicastat Fisher were from with not chronic for adverse the hypotension. 5, increased the treated the with are statistically pre-dose 10 recorded dopamine/norepinephrine ’ dopamine/norepinephrine nepicastat. none s the drug-treated Nepicastat LSD conversion days the heart the with events Then when with levels by of levels strategy after 80 were was the analysis pruritus. daily. failure nepicastat Concomitant a 160 mg significant. increased 105 compared one-way of drug used 10 in treatment generally groups of mg dose plasma to urinary days (CHF), dopamine For of treatments to for adjust One was variance ANOVA at analyze was to body of 8 to medications well or dopamine can days 100 the withdrawn dosing daily patient for those 160 ratio urinary ration to weight, increase vehicle multiple tolerated. mg/kg overall or had model, was norepinephrine. mg. doses receiving in with longer also plasma any levels performed was dopamine from a tended controls included of effects both plasma comparisons. had a two-way effect developed 40, The significant detected in the placebo or shortness 40 80, normal for to dose urine. were on or study for mg hydralazine or and decrease treatment, Basic ANOVA urinary body each at the and a in made (p increase because volunteers Compared 120 rash. which of < tests day, subjects ratio weights 200 breath mg 0.05). levels using body Two with day, and and mg the for of of of in 2014202047 11 Apr 2014 Nepicastat patient the three treated were the norepinephrine norepinephrine not increasing Example in due all embodiments, embodiments are suitable dopamine/norepinephrine cancer. chest subsequently [0383] [0382] [0387] [0386] [0385] [0384] doses, 8 suitable considered ratios to a patients diuretics. determined. pain worsening who with In Dopamine/norepinephrine In In One modifications 10 28 an studies including treatment creatinine It after in studies was nepicastat and serious ongoing will who had thereof. day 1 it related levels, levels, Occasional patient, receiving may is treatment be of CHF an received intended Dopamine/norepinephrine of placebo. congestive readily study increased adverse acute level, study be and to and or While and (patient congestive made and study disulfiram. levels placebo. dopamine/norepinephrine dopamine/norepinephrine adaptations MI with in which 160 cases apparent to were an event the patients Symptomatic dopamine/norepinephrine without cover and drug heart mg was in adrenal nepicastat invention of ratios required the cardiac reported receiving heart include: determined. orthostatic to failure such with 10 106 to departing one in mass day the alternatives, CHF, has failure hospitalization. arrest), the of orthostatic patients, in worsening as study. methods 80 ordinary ratios in been a hypotension brain possibly there from mg 1 30 unstable patients, patient ratios described ratios changes increased qd) Nepicastat day of have the and hypotension modifications, skill CHF levels rodents and being after study scope and applications who been Medically angina in were in in plasma one in the in changes in treatment plasma 2 related had 2 of the were connection treated reported patients was relevant the deaths: treatment in was the 30 a and dopamine 1 sudden brains significant determined. history dopamine invention of day described to patient, reported with with equivalents (one one the in arts drug study. with patients of levels death nepicastat nepicastat death that increased of of atypical was rodents in certain or levels, levels, events whom herein breast other 6 was in any and As on an as of a 2014202047 11 Apr 2014 position may more behavioural/physiological behavioual/physiological the was proposed behavioural/physiological Example of in effects claims. controls drug-treated administration same groups drug administered status animals acutely dose [0389] [0388] [0392] [0391] [0390] 8 a animals rats. administered be or of irritable dosing separate of of Nepicastat Prior In In The of vehicle physostigmine to included (400-480 had each 29 dose drugs study, The pilot pilot 8 male were delayed-matching-to-position rats animals schedule. a acutely to than animal weights of of mean studies administration. study, studies, on Sprague-Dawley received the within observed nepicastat nepicastat twice pilot g). controls short-term (30 commencement displayed to increase at The administered of physostigmine of The groups daily 1 nepicastat the studies , or the vehicle effects repeated 3 effects changes. ‘ when animals blind weights (100 spirit and at animals 100 (06:00 or in a a of 24 working dose body ’ mg/kg mean following rats were handled of or and by 8 (30 mg/kg administration, h were repeatedly. of rats. both physostigmine and following an Similarly, were (n of of scope weight the and (1, 107 loss p.o.) performed independent = 100 testing 18:00 memory acute observed Observations during (DMTP) animals 8) monitored 3, p.o. repeated 100 in of mg/kg and within of body 10 and h) the drug mg/kg there the nepicastat acutely) establishing 1 for in were or nepicastat g. by invention repeated (0.3, test weight p.o. the with or rats. administration. 11 observer throughout 10 30 were an Drug p.o.) vehicle were day monitored is same days 1, However, observer mg/kg used the did of 3 study. no in administration treated or made as or (100 the weight 28 in (once administration. the vehicle overt aims to defined not 10 order maximum the g p.o.) mg/kg throughout blind examine at after mg/kg in delayed animals on study In range induce 1, effects the to of day was or a 3 by 11 to assess p.o.) separate latter and p.o.) assessing and, of vehicle the as days the tolerated 11) administered the matching also of the the the any 24 or Similarly, following to treatment any on using study potential repeated whereas became hr vehicle highest study. trained groups day study, overt overt were after oral the the the to 5, 2014202047 11 Apr 2014 physostigmine. were two may were reverse was vehicle treated with basis were b.i.d.) training, repeated reduced overt of induce of 3 found administration choices, (0.1 across [0394] [0393] [0397] [0396] [0395] mg/kg training, testing measures mg/kg, to indicate either of treated able behavioural observed dead or a As Acute Fifty-six In The with a determine a treatment data scheduled the to administration. the physostigmine scopolamine group). significant the in to a nepicastat 30 on were s.c., latency result vehicle, dependent effects with the changes from complete present may administration mg/kg day of male at 30 DMTP used. throughout whether effects 3 nepicastat of There be a delay 2 mm 30 to of pilot and impairment in was these DMTP indicative or in Sprague p.o. induced make repeated during the mg/kg measures pretreatment They (1.0 memory physostigmine, of task. 10 were treated (chewing DMTP and studies chronic 10 either mg/kg mg/kg choice study, and the of were mg/kg the Dawley impairment a no p.o. On of administration physostigmine with dose in and/or experiment. study physostigmine 0, in 70 the other effects the housed administration mouth p.o., responses p.o. rats choice 8, (cyanosis, time). group, the min highest scopolamine. of rats, 16 108 eleventh in were one attentional non-cognitive physostigmine b.i.d.) 1 or present which test of movements in in mg/kg accuracy. weighing and The 32 group repeated choice trained dose groups and sessions. The of were s tremor, at 2 day dose 0.1 were in nepicastat study of the p.o. of of of doses function purpose order accuracy 8 Another mg/kg examined of between to of nepicastat nepicastat animals of number In effects administration co-administered animals and of the was remember four head scopolamine included Changes to of addition physostigmine salivation). of experiment, earn of whereas 3 (1, toxic per in group of which 200-290 jerks, of scopolamine the mg/kg across convulsed the for 3, or the food trials cage in percentage to the final (3 10 physostigmine the DMTP of drugs. had changes the ataxia). the was reward. of of 10 and position or and which rats g DMTP scopolamine Signs scopolamine the at previously former 0.3 8 successive was animals higher on selected were 30 the task. received animals was animals day or the in mg/kg chosen of of Following beginning study of Repeated the each found 1 measure induced toxicity animals 5 correct a treated mg/kg would on in latter lever were been days only was Hbr p.o. that test fed the the for to 2014202047 11 Apr 2014 maintained this pressed. the previously As retraction the minutes the right then retrieve the were the be houselight food approximately approximately sample levers levers lever and ceiling conducted centrally [0398] [0400] [0399] pushed soon magazine illumination operant food chamber an weight trained magazine. contained lever resulted (the were Acorn Matching With Twelve of Noyes lever) magazine. as long. located A of the back with sample was presented) was on equipment. the were to response the inserted. until the light. A5000 in chamber a did press 85% 45 12-15 magazine Campden in randomly the by Rats illuminated. of 12:12 lever The the food to given house mg sound lever) not a the the partitions of The The position both retraction lever-press g boxes were computer Formula to and resulted hour their magazine rat magazine produce of and additional A magazine light and partitions was the the Instruments food flap to presented the response placed light/dark free-feeding were extending enable light incorrect training left fitted inserted Following illumination was on per ‘ in of A programmed a response light. were and modified ’ retraction the throughout attenuating food rat light were food. pressed food in into it during to right to per cycle began lever, operant the into used 105 lever 109 The pellet remained the pellets obtain clear weight. the a Water day. occurred. operant the levers of the mm so 30 a with magazine operant for of delivery sample next. (the with 30 the shells. Perspex, that the chambers and chamber. magazine s food This both from behavioural into was mm the Any intertrial to magazine on opposite Arachnid partitions session, box initiated This boxes. obtain pellets, amount light behind the levers, until lever Paul freely of A session. light animals reaching and response a with central The and light period the food Fray interval food remained lever (i.e., rats The light the was available. a delivery testing. of the software could pellet lever two all which 10 was A food pellet Control magazine reward. area. sessions were from positioned beginning session subsequent (but to the response s to retractable be remained extinguished (ITI), time-out was that maintained the began on A of same was not the and fitted initially The The flap, System a until retrieved. presented lever began were Either pellet grid one flap. food used illumination to at operant to in lever animals either training which inserted (TO) the levers 6 the drop front floor of initially resulted trained pellet a.m. Rats to the the Interlaces when delivery). and flap inserted the as control side period below rats as left boxes of to could and were were both was was was into two and the the the the 50 of or of to in at a 2014202047 11 Apr 2014 times trials were was was the the response twice the trial. between with the response immediately From habits computer lever commencement during any contribute constraint first correction The determined [0401] [0403] [0402] session sample four further intertrial used. decreased number flap a used to (correction in on After Session (i.e., high A Following Following which a be types the within occurred, a press correction block that trials to on Due right in on length always sessions. inserted such a degree depression followed the of response all 25 interval the non-correction the to in of occurring to were 30 of of trial. correction data that onward plus subsequent 10 a trials was subsequent poor 24 26 responding houselight a the 16 s of 16 procedure (left new s of sessions, sessions used analysis. On the trials. and, a non-correction) A accuracy. increased lever began. trial performance to the (0, trial of limited the Session trial. after or right the to the end over trials 4, block, that right) on time-out evaluate sessions. sample ‘ with trials was 8 correction on the either was sample of and The The which the Such to was or hold was On 59, either the animals as each 70 the used extinguished. lefthand 16 same next lever at (i.e., not a delays Session lever, scheduled the was the period minutes. recorded, a a the during lever s 0-16 0 110 the Only delay correct throughout ’ chosen delay) trial 8 s trial inserted percentage sample trial. used the 64 were (immediate), sessions left the of s levers and following s 25 51 delay was Session was occurred first 0, delay, or choice such was These choice delay (i.e., performing but presentation a of 8, lever as right reinstated variable A were counted (Session (Session this the trial 16 only the semi-randomly of that the however, period, 30 correction 59. levers occurred). and incorrect was lever correct 4 56 4, sample and presented ‘ if of correct s the times; 8 32 rats These delay the ITI the randomly 51-58), as all following or the 25-50) and were the of data s choices. this rat matching subsequent 16 lever was were incomplete completed choices the ’ twice trials trial achieving session interval lever) rats Each did s inserted collected as delay the a determined levers delay. initiated was used. was the delay prevented determined not were the on time intertrial-interval was was was to was and semi-randomly sample terminated a make end more on training. 50% following These position of The and left selected presented on an omitted not before the interposed trials of up incorrect the trial with correct). a order position than did the used lever to by choice choice delays non 64 task The that ITI. and and and and not the the the the 24 of as in 8 s 2014202047 11 Apr 2014 nepicastat were Vehicle/Vehicle performance percentage minutes testing. used the (n=7), (n=7), semi-randomly collected correction sample seven group [0404] [0405] vehicle 4 2 7 6 5 3 Group 1 during run, groups nepicastat received At The nepicastat lever prior The for the (1,3, and 6:00 of treated the data the during animals rats nepicastat nepicastat nepicastat nepicastat Vehicle Vehicle Physostigmine (0, Session and (percent correct to non-correction assigned final am 0.1 10 non-correction 30 (n=7), 8, the collected received the 3.0 animals or 16, mg/kg/Scopolamine, and the mg/kg test responses; final in 30 60-69 performance mg/kg/Scopolamine and correct, 11 30 3.0 Vehicle/Scopolamine 10 1.0 6:00 to the mg/kg). session test during consecutive oral the mg/kg mg/kg 32 of mg/kg mg/kg and Vehicle/Vehicle trials pm s scopolamine following trials session. administration delay) 2) (Session response analyzed Due of sessions completed. the of the only. to Ill latency the days consecutive the and 7 Thus, V Vehicle/Vehicle Physostigmine/Scopolamine nepicastat nepicastat nepicastat nepicastat (0, Session 70) 68 in latency groups ehicle/ choice (n=7), of HBr 4, high Phys/Scopolamine group in and the of between the (n=), The 8, the which either 70 and degree S administered 69, present present such response copolamine nepicastat former received 3.0 30 drug and 1.0 10 nepicastat days the 32 all mg/kg/Scopolamine mg/kg/Scopolamine mg/kg/Scopolamine mg/kg/Scopolamine vehicle, performance that s of number treatments 0, but experiment delay) DMTP two during choice 8, the and a the 16 s.c. 10.0 dependent s.c. (n=8). groups physostigmine 1.0 Vehicle/Vehicle 3) and which of accuracy study injection 30 the mg/kg/Scopolamine mg/kg/Scopolamine administered of were: 32s trials were minutes a total Sessions response include measures delay displayed of completed): matched number (Phys) saline was prior treated 1) to 60-69 to were also the the the by on 30 or of to 2014202047 11 Apr 2014 the Vehicle/Scopolamine blocks was minutes percentage number of included one-way one-way during are first expressed suspended 6:00 30 sample assess complete followed analyze subjects conducted SuperAnova [0407] [0406] [0409] [0408] the mg/kg, animals f0 dissolved pm. included. 5 Blocks the (Block All In Physostigmine Two-way task, drug of blocks prior percentage factor ANOVA in by ANOVA order trials as trials supplied separately Scopolamine in overall of the the in all a base software. treatment distilled to correct 1-5. post 1-5), of in these analysis animals to and completed. at statistical the weight. analysis drug increase saline which Also, effects was hoc by each although delay of last for two groups choices sulphate water Roche) sessions testing Alpha were Dunnett of correct followed HBr each and was test of groups because of of (0, percentage the tests the and received figures included the variance was (0.1 injected session. conducted block and were and power 8, (1.0 (Session ’ choices s four injected 16 were drugs by were test, set mg/kg, on response mg/kg, administered of the for or and the a to the in of in 112 delay data. (ANOVA) when Physostigmine combined two-tailed the 32s) on 0.05 the conducted at 60-69) and correct scopolamine sensitivity animals a same in supplied supplied each ability data volume analysis latency. a throughout. Significant periods appropriate, as response volume treatment the delay. the choices collected p.o. for with Dunnett to in by data by of of on within-subjects of twice were The the perform (scop) the and of Sigma) RBI) number the A 1.0 latency. drug were interactions and a for was Animals purpose 2.5 on significant statistical number ’ nepicastat s Macintosh daily ml/kg. excluded and Vehicle/Vehicle latency test the t-test. Day treatment ml/kg. collapsed used was the of first nepicastat day starting trials These 1-Day which of of delayed to administered All analyses, to were factor One-way were main f0 statistical were animals Scopolamine from analyze respond completed. computer as into drug days 10 at were analyses followed recorded. the effect was (1, dissolved 6:00 matching of analyses two-session during of doses that the 3, unable the between- ANOVA and for analysis used am testing, s.c. from 10 mean using study were were each by HBr and and the the are To 30 or of to to to a a 2014202047 11 Apr 2014 with treatment nepicastat treatment this main of of impairment Dunnett F(5, F(5, of Block significant, group. and significantly failed significantly delay choices delay-independent fact, affect dependent [0411] [0410] [0414] [0413] [0412] nepicastat trials correct 40)=3. 40)=2.1 block 32 during either interaction to effect 4 response In During During During The Drug ’ revealed of s completed. reach s also X block of delays had choices 325, test measures the 3 F(5, 15, in this effects delay testing. differed were of or impair treatment failed test, a choice statistical the block at block 45)=1.717, p=0.01 latency 30 longer 2, block drug failed a the found p=0.084; able first or interaction. nepicastat impairment F(5, mg/kg of significant during to This response 5 from the accuracy. 32 3 drug of treatment to block of affect 45)=1 latency to did 3, or of a significance s latency training complete testing apparent reach of the for statistically number delay p=0.150, not this testing treatment nepicastat, the of and .533, vehicle F(5 the to main latency significantly in Subsequent block testing, statistical Two-way number nepicastat to perform found revealed only physostigmine ,40)=1. choice 0, effect p=0.1 the trials of perform there effect in of 8, treated on trials 4 113 significant animals with the block that the and at 16 99. out of 403, was percentage the accuracy. significance. was ANOVA all drugs that of trials one-way induced test. and affect in a completed, of only the group. choice 4 slightly of drug choice 7 a the (p=0.056). treated only the 32 p=0.244; trend had completed, Although ANOVA had of the the group group treatment, revealed a s the four response ANOVAs Drug no of ANOVA no response marked more group number the delays, towards with A animals correct F(5, significant significant delays. treated effect post treatment physostigmine the Drug revealing marked 45)=0. F(5, treated a F(5, physostigmine than dose- respectively. of but effect significant in on a conducted hoc choices at treated 45)=0. with treatment 40)=2.259, decrease trials the did percentage the 701, only effects Dunnett effects in and with did was DMTP physostigmine. a the the drug 319, completed with p=0.625, delay-dependent significant not approached, the not drug vehicle 10.0 at groups in treated on on did, ’ p= significantly treatment s displayed A 32 the the 30.0 task. statistically percentage test any of mg/kg treatment 0.899. however, p=0.067; post s 0, number correct treated treated during during mg/kg on delay; of group Drug 8, drug hoc but the the 16 of In X a 2014202047 11 Apr 2014 the nepicastat X Vehicle/Scopolamine with not p=0.003. the mean number respectively, Dunnett F(5 delay, found effects groups. scopolamine 30.0 found groups able scopolamine, administration accuracy except [0415] [0419] [0418] [0417] [0416] delay ,39)=3. vehicle conducted. choice to mg/kg scopolamine percentage that a for Nepicastat Nepicastat was Due Many The complete in F(5, of ’ s significant interaction and With the treated 018, trials test the treated scopolamine significantly accuracy of number 39)=0.327, to completed and the of Vehicle/Vehicle nepicastat the made of DMTP conducted fewer It p=0.021, completed, the trials of scopolamine response only groups is when small alone: animals. and group correct of and interesting animals treated in than more at 2 test. trials physostigmine trials number administered reduced, rats p=0.894; the subsequent significantly plus each showed both for at 4 difference latency F(5, choices treated subjects Vehicle/Scopolamine completed group the It at did the correct scopolamine of HBr. of and to 45)=1. each is of the the impairments 32 not F(6, 0, note, the notable data subjects F(5, or collapsed with Only one-way four of s 8, animals in perform alone in any 692, impaired Vehicle/Scopolamine 16)=8.801, delay 114 choices did 39)=0.825, the however, were the by 16 percentage 30.0 delays. of 1 performed p=0.156, four that the not and does other in rat the found not treated mg/kg across ANOVAs the in the treated animals delays. percentage significantly the 32 animals not than percentage subjected that groups, p=0.001. In scopolamine delayed group s that p=0.539; during delay of appear fact the of with well the delays, with any nepicastat correct in four both at in n further two to all 30.0 matching compared induced < to the Block the of 10.0 to in of 4 of the delays. affect of respectively. animals ANOVA. the induce which correct F(5, occurred correct treated other choices 0, mg/kg mg/kg the statistical Vehicle/Vehicle the plus 5 8, 10 39)=1. of memory response scopolamine scopolamine to to 16 seven memory A and in the groups, choices choices, scopolamine of animals of the sample at in t-test and the In 188, nepicastat nepicastat test. the only analyses all group and addition, group A 32 impairment the comparing latency the relative enhancing 30 t( test post p=0.333; four the s 9 in )=4.15, treated treated treated groups choice mg/kg delays group given were were after 32 rats, plus plus hoc the the or to s 2014202047 11 Apr 2014 blocks memory model to treated the the perform nepicastat observed. observed only impairments However, or in induced Block disrupting animals (e.g., able showed accuracy. “ scopolamine days directly accuracy effect accuracy [0420] [0421] non-cognitive the which physostigmine cognitive animals to when and MK-801, vehicle 3 in have Notably, Nepicastat of group. accurately no of on were typically impairments the at the at the actually in two testing. and effects. testing impairments the short-term the The the shown in in treated the animals DMTP disruption treated performance still ” rats physostigmine retention scopolamine). 8 choice the 0 The actions animals by vehicle and induce (Days s induced produced could perform receiving showing Physostigmine this 10.0 However, animals. delay the groups group were task 16 in accuracy or of in induced profile. s 5 fifth interval mg/kg not treated treated impairments working choice delays the & choice and displayed co-administered significant the of treated at a a 30.0 6). be It block compound. treated Nepicastat by delay-independent delay-dependent the the 80% DMTP is at nepicastat by The analyzed and is accuracy with control Many accuracy mg/kg possible the the memory. 32 did vehicle long. with scopolamine, choice of results a a s groups 0, dose- in fifth not 115 significant 10.0 task delay. training higher compounds 8 of choice the animals had Few and improve that due at suggests group treated nepicastat scopolamine block accuracy at mg/kg from and The the that highest a The the 16 nepicastat to choice compounds small memory accuracy nepicastat 0 delay-dependent impairment animals were a s an impairment were appeared of the shorter s delay-dependent performance delays. scopolamine animals of which delay, that effect at testing dose effect and small nepicastat accuracy able impaired the HBr impairment the which is are delays scopolamine a have that may of In that capable 32 on to at to tendency the number (0.1 compound in in sufficiently contrast, 30.0 perform the dissipate latency s may have is challenge choice have on been choice than vehicle delay. and relative showed impairments mg/kg) observed 32 nature mg/kg any of be with been of to show claimed selective reversing any to s at that accuracy masked the accuracy of impaired across subjects Thus, may treated delay complete the motivated to and test of absolutely the tested of 100% of were DMTP impairments at the these 32 nepicastat nepicastat on be all the treatment to in a by was memory the some s relative vehicle control able ceiling day choice impair during in choice choice delays in acting delay, drug- other other trials task. five this and not the no 11 of to 2014202047 11 Apr 2014 the variability variability trials treated treated which training nepicastat, the Although were block independent: complete longer The secondary significance. correct administration This animals extra cases consistent groups deprived [0423] [0422] 10.0 animals chronic animals mg/kg separated observation in feeding completed of was reached with Physostigmine We showed animals. choices in during excess animals, training. fewer systematic performance to in in the apparent the found treated of 30.0 oral the appeared the behaviorally the Thus, may of 30 nepicastat statistical reductions and which trials impairment of during interaction data mg/kg. This data, were 100 dosing mg/kg that particularly 5% is with given have on The the recordings than mg/kg was the did consistent effect some however, to the of across observed Block physostigmine effects significance. group animals There took regime contributed develop total not additional the impairments in unexpected. toxic fourth term in p.o. was of the vehicle improve the during longer 3 body than choice was the of effects were number of these from not block of next recovered may during with tolerance physostigmine nepicastat animals also in training. food weight. dose-dependent treated the to to not the three performance trends 116 accuracy have the in showed of of It the completed of a the the first at analysis appears made, training, choice trend the other trials blocks in the to results from animals: disrupted variability last did induced Animals drug few this these In a end for groups casual induced completed accuracy significant not two on likely this of contrast of blocks during study the the when of of of variance effects reach training. and response a the trends initial blocks which their animals the 32 marked observation a had that in began was by it rats which of s induced pilot to number performance statistical between is impairment delay to daily over the the for physostigmine decline did were in the not of administered accuracy be treated loss to reductions initial the the experiment. study not the training. training observed given trials effects lose by showing of of suggested animals DMTP the approach significance. last and physostigmine body with stress trials weight, in than in first additional are of two obtained showed session. percentage in which nepicastat test. in at weight. treated these likely Due weight was and induced the completed. number All that the this blocks statistical in vehicle In second to delay of group food- to some more dose. food. more daily with with This This fact, loss the the by no be of of of to 2014202047 11 Apr 2014 may the to with the present unlikely, unexpected the task. the nepicastat treatment unequivocally test treated be of induced administration effective due and “ scopolamine able day during scopolamine appear after [0424] [0425] non-cognitive scopolamine compare employed scopolamine cognitive fact nepicastat MK-801, to to 11 showed physostigmine be 8 However, Block to Nepicastat Finally, perform with impairments in days that, required results that against dose- reverse days or which with finding 30.0 treated physostigmine compared for 3 changes disruption of higher induce treated whether ” and of and of physostigmine in the and with test. two the scopolamine. actions dosing. chronic the to mg/kg the testing this a appears physostigmine actually given DMTP using higher animals. reverse present delay-independent effects rats in delay-dependent choice animals It in test. nepicastat. groups. acute with is induced attention attention of of (Days administration. receiving that the to Physostigmine possible task dose produced nepicastat the could of the results. acute have or accuracy present It many did were scopolamine We 5 is displayed chronic compound. effects of by & or Further not and/or possible not specific treated administration, have that physostigmine If 30.0 other scopolamine, 6). co-administered motor/motivation This dosing a appear be that this 117 impairments delay-independent than a impairments of treatment The not mg/kg analyzed different motor/motivational The memory a did lack groups memory-disrupting that were scopolamine. research drug although higher previously results regime any to On animals not of nepicastat reverse of able the is effect an that of with due a dose nepicastat disrupting improve (see choice from and lower in in selective effect it scopolamine would to the final performance would to were is choice nepicastat attempted the results of perform of therefore choice a the interesting is In animals impairment accuracy physostigmine scopolamine dose physostigmine effects that block capable performance able addition, small not drugs, be and factors. accuracy effects of for may of accuracy. during have to to needed scopolamine the have can of of HBr number physostigmine in a could than such of reverse that perform be scopolamine training, which pilot novel in reverse nepicastat any tolerated reversing no In that masked (0.1 challenge the any choice the as may account on may historical to contrast, of of study) scopolamine scopolamine scopolamine are are two mg/kg) receptor of This any the subjects the have nepicastat determine the be that the probably accuracy apparent repeated by some animals did DMTP test effects for during had of due which is other other other were been it data and not the the on an of or to to in is 2014202047 11 Apr 2014 working pharmacological matched hydroxylase Example administration strain sequentially: examined. enalapril. SHRs. [0426] [0427] Series Series Series of Recently Male The memory. rats. 30 WKY/NCrI III II I Vehicle nepicastat nepicastat Vehicle nepicastat nepicastat Vehicle Enalapril Enalapril Effects antihypertensive inhibitor, SHRs/NCrl Furthermore, of mechanism, we the of BR have exhibited the compound 10 3 10 100 30 10 BR rats mg/kg treatments mg/kg mg/kg mg/kg mg/kg we demonstrated mg/kg effects these rats were also an (22-28 results with effective of used. explored on 118 nepicastat the weeks the suggest that Four antihypertensive angiotensin the cardiac old nepicastat, were series possible an at important hypertrophy examined the of potentiation converting experiments onset a activity role selective chronically of in dosing), for in enzyme effects SHRs were this acute dopamine substrate in and conducted were of studies the inhibitor the weight same also co β- in in 2014202047 11 Apr 2014 telemetry ventricles received After mean recover measurement obtain into off.) housed sutured and aseptic anesthetized [0431] [0430] [0429] [0428] Series cannulated 4 the blood dry groups At In Twenty-four Three conditions, individually for to a predose monitoring IV each were weights. 30 the nepicastat nepicastat nepicastat nepicastat nepicastat Enalapril Enalapril the at pressure with day days least of and start abdominal with series, collected, arterial daily pentobarbital values their an 2 prior of the in hrs weeks (MBP), was incision 24 treatment each a catheter systolic 60 30 15 1 30 30 1 to after for quiet blood SHRs mg/kg mg/kg musculature, always weighted mg/kg mg/kg mg/kg mg/kg mg/kg the before these experiment, heart was the room sodium start pressure, of were blood of (El) 6. parameters being last made a + rate + + + nepicastat (wet telemetry of enalapril with El El El In chronically 119 treatment, pressure (60 (HR), the the Series subjected on the weight), heart reversed mg/kg, experiment, skin midline. number were and/or and 1 transmitter I, mg/kg rate (SBP), was however, implanted the motor to and i.p.) established, light/dark enalapril and drug The of closed. rats lyophilized and diastolic the rats activity unit. motor abdominal administration. were 7 its rats with in Wistar (see cycle Each abdomen After respective each were activity. sacrificed blood (MA) telemetry below). for (08:00-20:00, rat Kyoto the aorta group randomly at were pressure was transmitter shaved. least groups The was The implants and (WKY) undergoing monitored. allowed rats exposed, 24 rat the divided (DBP), of Under lights hr were was was rats rats left for to to 2014202047 11 Apr 2014 was to were was weight/body weights procedure. performed. the weights instrumentation Bonferron Wallis interaction 26 additional Then control and as These for SBP, [0433] [0432] [0437] [0436] [0435] [0434] the a time end p time obtained used statistical DBP, a similarly level rat were A test All Both For For Data would one-way of were points was in and to computerized ’ the 2 s values body left of was was MBP, If then rats 10 adjustment The on analyze nepicastat weights. used commercially recorded treatment, no their be less measured ml/kg ventricular analysis housed MBP, or used in detected, ANOVA pairwise performed averaged weight overall HR, were than to monitoring each interaction overall analyze to and HR daily. and If and 0.05. to and expressed data group left analyze treatment a on each an were comparisons was multiple and a mass, hourly two-way MA. enalapril from by dosed ventricular effects overall series collection effects tissue was thy. MA performed were was adjusting expressed a Data ratios an and as conducted local effect A with comparisons were of used. for analysis ANOVA treatment wet similarly were means on standard on two-way one-way 120 of at treatment, mass system pharmacy. vehicle the weights analyzed was each the for as prepared each tissue If ± critical free of on (dry each an treated detected, with SEM. effect hypertrophy rat was errors time covariance ANOVA was these ANOVA overall (water), wet base and were and day, separately. day, respect in value used then point among (to weight/body Statistical tissue of water. rats. wet equivalents. then and collected and treatment the increase to made. with while using weights) were at with to treatment pairwise of continuously dry the mean all All each Each the main a groups significance pairwise SHRs). performed a in weights, every doses the Bonferroni changes covariate (SEM) weight effect time Series were Enalapril analysis effects comparisons numbers by was 5 were point obtained. min. day difference or while collect calculated. III and No from using of for not was a was adjustment. given (Vasotec®) interaction. and for of significant final tissue would treatment telemetry detected, Kruskal- pre-dose done animals data defined Dunn 10 for IV orally Body body from sec. dry the At on on be an ’ s 2014202047 11 Apr 2014 those nepicastat was most nepicastat maximal throughout nadir mmHg Fisher With 21 on study. little day expressed drug-treated day antihypertensive low (15, dose administration 8 [0438] [0442] [0441] [0440] [0439] 1 through and day mmHg treatment studies. observed average 30 recovery of of a induced ’ continued of s Nepicastat 5. For In In Enalapril second 21 At and the enalapril LSD decrease approximately day series at hereafter hr series at at the series 100 day 60 60 The mean groups hr after strategy throughout by 30 (data within 30. of administration study. mg/kg). mg/kg with mg/kg, 16 responses to IV, the the III, groups at was I mg/kg at of dosing Thus, blood and on lower not are 3 two 10 potentiation compound to 24 combined -29 although to studied day and initially II, shown). po) the the hr. mg/kg, adjust treated the the mmHg The compounds (n=6) -42 on pressure the Oral than 10 1, did compound Similar vehicle potentiation study. day p<0.01). on MBP group mg/kg, mmHg further. produced for treatment mono-administration the administration at with not induced consistently At day (p<0.01) 1 of multiple 15 with (p<0.0 rest). to 30 antihypertensive significantly that the controls 2, nepicastat (n=6) or did a on 121 mg/kg, induced In The a 30 maximal a a effects received was only not greater (-25 1). in large the day greater induced comparisons. mg/kg, onset MBP consistently MBP not lowered were presence mmHg of nepicastat a 3 at standard of a antihypertensive nepicastat affect small related of nepicastat and of (p<0.01). 100 peak was no nepicastat continued a made -20 the effects greater of at greater longer mg/kg, MBP observed blood of antihypertensive antihypertensive hr response mmHg enalapril to gradually error affect using enalapril 13, the at were antihypertensive 15 antihypertensive throughout effect to however, by pressure The 3 p<0.01). (two doses heart mg/kg on decrease and within Dunn induced a response was at lowered MBP was non-antihypertensive day (1 rats 10 of rate 1 slow ’ mg/kg), and on s tended mg/kg 1 3 mg/kg nepicastat The observed showed response effect, the and procedure hr throughout (HR) remained (p<0.01), any was El MBP and potentiation after response reached effect study. to (all of exhibited observed in (n=6) although gradual. the exhibit greater on on dosing the the tested doses of-29 with than low day and day co the 30 30 its or A ( 2014202047 11 Apr 2014 treatments, Although nepicastat nepicastat the but motor mg/kg) induce of II, enalapril lower (30 slightly (p<0.05). and left decreased effect consistently SHRs [0443] [0448] [0447] [0446] [0445] [0444] co-administration but ventricular compound and the 10 (p>0.05). activity on HR not mg/kg Nepicastat In Throughout The The Compared a co-administration and 60 decreased (1 transient the enalapril comparison, the at at in mg/kg) than mg/kg) enalapril affected pre-dose pre-dose i.e., 30 3 Series body did left (MA). and at mass and the Enalapril 30 not enalapril ventricular to at small body slightly 10 alone. at the I. weights 100 vehicle were of that and HR. 3 have (1 mg/kg 1 body body In enalapril study, enalapril - mg/kg mg/kg weight, mg/kg) tachycardia treated 100 100 Series not of (10 In increased any (1 were control of enalapril weights weights Series mass mg/kg, different mg/kg none mg/kg) were effect did the mg/kg), III, tended at with co-administration (1 387±1 of not 10 mg/kg) of IV, rat. enalapril the 399±10, group, within did on however, vehicle, the significantly of of (1 the mg/kg from 122 affect the to body body 1, mg/kg) not Although nepicastat the the SHRs drug exhibit 415±12, and groups 2 at enalapril 389±6, affect at weights significantly weights HR, rats rats hr treatment least induced treatments nepicastat 1 (p<0.01). after and mg/kg lower reduced the treated of treated treated 407±4, treatment the in 389±9, (30 alone nepicastat of enalapril in dosing. compound the cardiac greater of the SHRs did HR mg/kg) with at showed In decreased the and with with (p>0.05). left first rat. and not than 15, Series with SHRs ventricular 415±12 (p>0.05). In (1 nepicastat increases hypertrophy (30 401±10 regress few 30 vehicle, vehicle, at the mg/kg) Series a or enalapril and IV, mg/kg) significant 10 with hours (p<0.05) group the g, mg/kg the 60 however, g, III, in Treatment respectively. (15, nepicastat and enalapril, enalapril, mass respectively. mg/kg combination, after hypertrophy, body significantly treated observed none at nepicastat 30 or tended effect in 1 dosing. had weight and effects mg/kg on of Series with with at and and the the no on 60 to in 3 2014202047 11 Apr 2014 Nepicastat these nepicastat nepicastat mono-administration magnitudes the or Daily heart induced 346±8 administrations g, contrast, dose (1 30-day lasting effects, comparison, study. 3 experiments significant effect [0451] [0450] [0449] [0453] [0452] (p<0.01). respectively. mg/kg) 100 treatment dependent. deaths rate, was treatment mg/kg, g, Four In The The The antihypertensive study. the At an respectively. enalapril was at (15, groups at effects gradual co-administration 100 motor antihypertensive effects pre-dose of were pre-dose deaths 30 in the The 30 of however, also The 30, mg/kg, antihypertensive spontaneously nepicastat. of mg/kg mg/kg of angiotensin undetermined, antihypertensive on treated activity nepicastat and of at and seen potentiation nepicastat were heart of 30-day body 10 body without (n=6) 60 nepicastat reached exhibited at effect enalapril mg/kg observed with mg/kg) doses rate and weights weights effect with induced chronic at converting at (p<0.01). hypertensive and were nepicastat left of its but 15, induced of effects 3 slight (1 nepicastat and (n=5) the of-20 effect with peak in 15, and ventricular it 30, observed. mg/kg) oral of a of the appeared enalapril antihypertensive 123 30, peak bradycardia and enalapril 10 the induced of were to the a was The administration 4 enzyme and at transient -42 mg/kg -30 series rats (30 antihypertensive 60 rats did rats 3-10 modest potentiation observed 60 The mmHg unlikely (1 mmHg mass mg/kg mg/kg; not (SHRs) a were treated mg/kg, mg/kg) of treated inhibitor (n=6) greater mg/kg during groups tachycardia. induce 30-day were but 365±9, were on effects throughout n=6) that of for with although did with tended were the day with was or antihypertensive nepicastat that evaluated significant enalapril these the 357±6, induced treatments. effect not enalapril awake 371±8, radio-telemetry of 3 enalapril received observed detected vehicle, rest (p<0.01). to nepicastat affect deaths these Co-administration the of exhibit 363±6, hours a on of 361±7, -20 (10 in study. greater antihypertensive at effects blood nepicastat blood the throughout were throughout alone enalapril, four mmHg The mg/kg, of 1 slower Comparable by effect. 347±8, mg/kg, and study. the related causes and Although implants. pressure. pressure, series were enalapril and on rat. 369±7 long- heart at n=6) The and and day not co the the no 30 of In of of In to 2014202047 11 Apr 2014 mass, the rate to mg/kg nepicastat nepicastat vehicle. hypertrophy, Example of isoflurane on enalapril and consisted autonomic 30 day statistically formulation greater significant pg/kg), SHRs [0454] [0458] [0457] [0456] [0455] nepicastat. intravenous motor left and than treatments. 2 than however, Nepicastat A Beagle In Treatment females. of ventricular anti- and The 100 31 enalapril activity of the (1 study (15, gas. agents (30 nepicastat nepicastat effects different was 1 acetylcholine mg/kg, the hypertensive causes four mg/kg male 30 dogs mg/kg) doses Before was then was Three was co-administration in and significantly series Each with (1 mass on and anesthetized, were of from not mg/kg) administered n=8) of without detected. conducted treatment 60 through dosing the with death rats 1 nepicastat of autonomic of mg/kg). animal dose female, that administered effect the left experiments, (10 or were a alone. were with of related causing non-antihypertensive nepicastat reduced SHRs. an ventricular pg/kg), alone. was enalapril to instrumented and and found to undetermined, test intraduodenal of at agents, evaluate In each the to surgically the a 124 The 30 formulation, blood any These any single were in four greater nepicastat nepicastat-treatment (1 two (30 animal (n=6) effect norepinephrine groups hypertrophy of mg/kg) reflex deaths the pressure effects, the evaluated. mg/kg, intraduodenal dogs. compounds effect instrumented but of and and effects cannula. treatment treated the (15, tachycardia. occurred alone. co-administration dose appeared the however, in 100 n=7) on average 30 observed SHRs blood of of with regression A (n=8) (3 (n=5) and groups, nepicastat The alone doses enalapril during group single pg/kg), unrelated while over blood pressure were nepicastat 60 significantly vehicle-control mg/kg in Co-administration of mg/kg) did consisted no the the bolus not pressure of SHRs. anesthetized on isoproterenol 0 (1 significant on 30-day course not responses to (vehicle) hypertrophy dose left mg/kg) did and the responses dose and regress ventricular of decreased responses related Although treatment not one of period was 2 of the males had group to effect or have with with (0.3 test not the the 30 60 of to to in at a 2014202047 11 Apr 2014 was weeks toxicity ml/kg vehicle water. powder weighed Farms, orally of housing applied compounds completion autonomic each Chow® animals cages assigned assigned for pg/ml), [0459] [0464] [0463] [0462] [0461] [0460] toxicity. the animal evaluated once identified of before A At After Dose The duration On Inc., and by with isoproterenol was study were the even 10.3 a to vehicle-control the the agents the of daily. 60 dog dogs was a on In North offered treatment vehicle. assignment selection dosing. to transferred the time numbers. in nepicastat-treatment day mg/ml vendor. a and hemodynamic of was 12.9 with 60 dogs, 1 was experiment, Clinical were -month of use. Rose, of mg/kg. once those selected kg treatment, the During (6 dosing, environmentally nepicastat The was a groups; evaluated and The On single to pg/ml), into New daily study group study, signs constituted considered based the each females animals the because sanitized each a York. parameters. and study, oral the 60-mg/ml number, males of of and acclimation doses solution. day on approximately toxicity group dog water dogs dose 1 weighed data were acetylcholine the of male Each it 60-mg/ml 125 controlled. were healthy cages was of is dosing, were of animal was dogs of from 5, suspension commonly were acclimated The and 400 euthanatized dog 2 Beagle 20, period, assigned every 8.5 provided approximately males were two were mg/kg present total was 1 aqueous number, or nepicastat to 1, female The (200 other 80 Π.2 2, housed studies dogs the identified and dose was used. used to and mg/kg odd resulted ad cages at general kg. and pg/ml) solutions laboratory 2 week. and were libitum. were prepared 80 of 3 to females formulation individually with numbers removed 14 hours mg/kg/day. were The were tattoo nepicastat evaluate uniquely in administered to condition obtained were Purina nepicastat. transient of 16 dogs cleaned administered by after were conditions number. norepinephrine months and prepared from mixing the retained Certified in by were administered dosing. administered from of females effects stainless clinical daily the an each In old. 1 The nepicastat randomly ear at study. ml/kg in Marshall an potency to and Canine animal least At of sterile Males tattoo room acute signs were dogs steel test (60 the the of to 3 1 2014202047 11 Apr 2014 mg/kg, the recorded nepicastat. was to Arterial to use rate maintained mechanically table ketamine protocol. needle PO Each and anchored administration. filled solution. advanced electrocardiogram artery directly [0466] [0465] [0468] [0467] the an 2 dog in the analyses. of selected external animal cannula pyloric on was measuring A A The The IV) approximately skin electrodes blood being into before into single midline (10 The top Food into with and cannulated left dogs of Dose the being ventilated mg/kg) sphincter. pressure because tip position, samples was of evaluated the the removed was femoral intraduodenal dosing isoflurane duodenum The were of blood a laparotomy (ECG) abdominal vena volumes advanced were the evaluated withheld circulating and needle transducer were 1.5 the using surgically (data the arterial gas throughout vein cava A from was placed for diazepam L/minute). gas oral needle cannula levels through withdrawn were assessing not through a was incision from euthanatized for was was the dose was (1.5% polyethylene cannula route warm- tabulated subcutaneously and instrumented administration study. was cannulated performed and calculated the withdrawn ’ initially of s the (0.5 an to stopcock the systolic was is vehicle inserted the animals anesthesia. Rectal 126 from was intraduodenal 2% experiment. water mg/kg). a needle reapposed. data in by proposed of anesthetized advanced the tube this and and and on or body an tidal from overnight pad was into according are arterial nepicastat of and overdose report). the the diastolic the to Each filled not volume the exteriorized autonomic temperature to the A into clinical monitor duodenum tip basis into cannula. presented duodenum maintain surgical cannula animal with before incision by of At to the the aortic of formulation delivered of the injecting procedures the route lumen 50 sodium thoracic a agents. individual was polyethylene surgical for outside The was plane end was pressure in body U/ml standard site, and blood this of placed of for intraduodenal monitored isolated in pentobarbital (IV) the of aorta administration was the of each temperature oxygen The report. of described instrumentation. test anesthesia pH, were tip heparin-saline body the limb on a cannula administered and left experiment, formulation of just tubing mixture PCO a abdomen, recorded. at External a only surgical coupled weights femoral lead saline- caudal a in 2 route , flow (300 was was was and and the for of of II 2014202047 11 Apr 2014 with between physiological necessary, norepinephrine blood minutes pH, preparation; were which Approximately bolus approximately acetylcholine change administration gas agent. agents dosing, animal directly after [0470] [0469] [0474] [0473] [0472] [0471] analyzer PCO2, the 3 used directly the ml pressure Aortic Approximately Following The The Systolic, was was from on after first the each of blood to only a and responses autonomic dosed these repeated water. were administration baseline). polygraph dosing ranges intraduodenal blood dose. bring (10 into 15 (baseline) 50, administration. PO just diastolic, for samples seconds) data manually with pg/kg), 2 surgical the 110, Administration pressure, with (pH Following were arterial assessing to with before are 30 agents, vehicle duodenum recorder. norepinephrine Systolic, and = test were minutes and not evaluated and using 7.43 were approximately of recorded instrumentation, blood cannula formulation. 170 heart presented and acetylcholine. at norepinephrine mean the withdrawn. each or to The the the minutes administered diastolic, nepicastat. 7.50 The after using rate, at pH level of femoral before administration responses onto aortic time the was 127 the blood and were the in and and the of agents the this after 10 of PCO time second dosing and flushed ECG blood Heart vein PCO2 anesthesia characterized intraduodenal pH, polygraph ventilatory report. peak The minutes (3 intravenously to dosing, 2 mean was cannula of parameters = pg/kg), PCO rate, isoproterenol pressures dose and administration levels 22 of response with peak repeated an aortic to 2 between , approximately ECG, administration and volume chart and 27 agent, with isoproterenol within adjustments by 3 pressure cannula. mmHg). the blood were were PO to evaluating at ml by and approximately approximately the 2 was the and stability administration each approximately values bolus of continuously blood of evaluated pressures, cannula approximate acetylcholine, acetylcholine increase 1 Immediately vehicle 50, of agent ml/kg (0.3 were the injection from gas the of 110, was pg/kg), mean just 20 the parameters (maximum 10 autonomic given and the made, for recorded solution. and minutes of minutes flushed normal time animal before blood blood aortic (over were after each each each as and 170 at if a 2014202047 11 Apr 2014 norepinephrine norepinephrine behavioral present. related postdose incidental. of Example isoproterenol intraduodenal sodium characterized cage and cm system and eight class (DBHI), doses evaluated [0476] [0475] [0480] [0479] [0478] [0477] peak x 3 water was have 45 under photodetectors of No At Adult Locomotor The Surgically (San differences pentobarbital pressure cm responses on 32 lightly before autonomic effects the testing. were treatment-related No a effects x locomotor Diego normal male by 20 were dose end were treatment-related were covered decrease evaluating dosing cm; allowed on instrumented, activity of to Each CD-I between of Instrument of present. locomotor of (approximately light/dark w acetylcholine spaced acute agents 60 present. the activity lesser x and in animal (ICR) for mg/kg 1 ad was experiment, clean the x intraperitoneal approximately predose each equally h) libitum. differences magnitude (norepinephrine, No Co.). activity. monitored diastolic cycle in was mice placed of cedar differences anesthetized In isoproterenol mice. were treatment-related nepicastat. only 300 vehicle-control and with along Each (30-40 128 bedding. within All each aortic present. than mg/kg, used postdose between It in lights administration station 1, the animals has between an g dog 2, the a once. beagle blood on and length Blood isoproterenol, automated metal and IV) been on predose study was consisted responses predose acetylcholine between and 3 were pressure dogs, differences predose frame dogs suggested hours pressure of euthanatized day removed the responses; of naive 14 were the and containing nepicastat, of after 0900 ) wall. just to station and were and responses a that autonomic postdose postdose to administration. clear from between dosing. administered before hr postdose acetylcholine) housed The drug this by and activity compounds the perspex 3 a an bottom was photoemmitors and 2100 DBH to treatment No study. responses responses predose responses in overdose agents intravenous monitoring considered at treatment- groups hr. cage inhibitor a the of of single Food were were each time this and and (25 of of to to to 2014202047 11 Apr 2014 minutes. period, nepicastat were the ml/100 treatment, using or of intervals hydrochloride, Dunn Following each (both counts each (both sonicated. comparisons. effects a significant (defined [0481] [0485] [0484] [0483] [0482] one-way testing). vehicle salt placed time ’ time p<0.0 p<0.0 the s g. A When In For The and existed. weight motor procedure as overall 1-4 All repeated the time (10, and treatment point interval this ANOVA nepicastat, a 1) 1) mice ambulations, individually break For doses while while comparing overall activity was returned the 30 interval habituation Pairwise (i.e., revealed were ranked and SKF-102698, dose used. of and measures revealed reported the the effect was model, 2 was 100 the the placed immediately and consecutive treatment treatment was Fisher into data the there performed comparisons mg/kg), that dose monitored first was treatment period, 30 are the one cocaine there (nonparametric two-way in ’ s was there detected mg/kg. range 120 represented the the LSD of cocaine by by SKF-102698 was photobeams) the testing a the to time time to minutes were by for to dose was significant 129 were same strategy analysis activity see Compounds a at mice the time 180 interaction interaction group significant time 3-100 significant room range at as vehicle then cage. technique) minutes. interval of were which the intervals cages (30 to of had for mg/kg testing; at overall performed free was Following adjust variance least and were dosed each group was was effect of significantly and interaction. overall base, Activity and 100 to 5 30-100 the not not 1 effects animal all allowed administered and for hr test with intraperitoneally for mg/kg), was intervals, except at significant. significant. (ANOVA) p<0.01) prior a 6 treatment the for both each 60 (i.e., respect counts were for mg/kg. dissolved to At problem the greater minute to for treatment cocaine treatment time explore the each testing. recorded while in overall cocaine in and to was effects last The The any, a interval pre-treatment time total in both For volume ambulations of with (30 for 60 no analysis analysis performed dH effects treatment The and and in every multiple interval, minutes 30 cocaine activity activity at mg/kg) 2 overall O which either using mice min. time time time of and 30 of at at 1 2014202047 11 Apr 2014 mg/kg. three mesenteric nepicastat were nepicastat randomly mg/kg weighings or in hypertensive hypertensive Example interval, did groups and counts group). administration, study, administration shown examined. [0487] [0486] [0490] [0489] [0488] a the volume not 6 dosed (all treatments were ambulations to and Nepicastat Acute compared In Male had Cocaine In cause as The 33 p<0.05). inhibit contrast, at were assigned (n=8) contrast, These a compared artery no of orally used. 10 spontaneous significantly rats. remaining rat dosing of 10.0 any significant animals mg/kg, dissolved was the except to for 1 data brain were at there mg/kg once Changes ml/kg. significant vehicle for Animals acute to effectively enzyme the to 1 with suggest one were 1 any harvested, and for 31 vehicle were cortex a mg/kg, following in or administration hypertensive greater effects day control rats the of day of vehicle in 10 10 anesthetized in were no that change the the mg/kg norepinephrine the demonstrated for continued dopamine-3-hydroxylase and control. mg/kg 25, or significant number on these at nepicastat-treated weighed, allowed a mesenteric following 7 (dH where 18 vehicle mesenteric any total or nepicastat 130 in rats 2 was days. DBHI O) of 25 time Similarly, with total of receiving activity nepicastat n=9. (SHRs), to food differences ambulations prepared days group and as yield treatment interval. are halothane, artery At and activity a were artery and analyzed On devoid with locomotor 4 of or oral dopamine SKF-102698 16-17 oral groups hours and day at water deionized examined. in ambulations or in vehicle, doses doses after groups: administration of at terms decapitated left inhibitor 7, either weeks the from time motoric after ad four or stimulant ventricle that of levels 7 ambulations the libitum. water of the 1 intervals 3, 24 at oral and the old hours could mg//kg the nepicastat at doses 10, SKF-102698-treated actions total rats in and at last at 25 any administration at in 30 free the the with after be 10 (n=8/treatment activity the Animals the of spontaneously treatment 1-4, days or or time administered in ml/kg. spontaneous onset at base. 30 dose one 100 cortex compound mice. 10 has but any and interval of counts mg/kg mg/kg of of of not were been time Rats oral The and 100 the the the 30 of 5 2014202047 11 Apr 2014 measurements. mesenteric mg/kg not the treatment treatment rate. on comparisons days) catecholamine (p<0.05) differences dopamine/norepinephrine dosing, dopamine/norepinephrine dose dopamine/norepinephrine dopamine/norepinephrine (p>0.05) dopamine/norepinephrine significantly greater [0492] [0491] [0495] [0494] [0493] the significantly vehicle group Each means, using dose After In In The Statistically, (p<0.05) the different differences the (Figures the groups variable were artery in group ratio. of animals 25 (p<0.1) 10 cerebral a norepinephrine adjusted mesenteric levels. The each non-parametric days (p.>0.05) significantly compared mg/kg and than (Figures (1 6-8). was The last the treatment or of lower in cortex cortex, sacrificed for control analyzed blood ratio three treatment, 10 dopamine dose ratio ratio ratio ratio There differences artery, ratios different 6-8). to mg/kg norepinephrine from levels. after (p<0.05) of vehicle. pressure in treatments at compared of with in group was one-way the the separately. day compared this the after on this seven cortex than nepicastat), control levels 1 None a 10 7 in 1 group mg/kg day 131 higher had measurement group with slight both Norepinephrine mg/kg sample mg/kg control, days to were levels analysis of compared the were the 25 significantly to was levels For 7 nepicastat were the significant compared of nepicastat days vehicle compared the nepicastat size, f0 or were treatment, figures nor parameters of also performed mg/kg harvested, in and dopamine of vehicle was to (p<0.05) were significantly to the group. also control dose variance levels to 6-11, a (p<0.05) made dose nepicastat vehicle higher at the the significantly dose norepinephrine the using measured group group, used each weighed, *, vehicle in group, dopamine in and the There on 10 p<0.05 group either the (ANOVA). dopamine mg/kg day increase in Fisher experiment-wise time (Figures (p<0.01) 25 for dose. but were after 1 either group. 22. were days was mg/kg and dose and period (p<0.05) blood ’ levels there dose s seven no levels LSD in 9-11). analyzed significantly significantly **, greater (p<0.01) of group levels The significant the group nepicastat in were (7 P<0.01. Pairwise pressure the strategy days higher the or cortex cortex or were error than two had and the for no 25 10 of of 2014202047 11 Apr 2014 nepicastat may hypertensive on embodiment are suitable dopamine-3-hydroxylase claims. embodiments, [0497] [0496] the suitable be contrary, Nepicastat, It included modifications will compared and thereof. rats be it it is may within readily (SHRs). is not administered intended to be While intended and 1 the made mg/kg, apparent in adaptations Greater spirit the to the without to orally cover therefore invention and limit to inhibition cortex one scope such for 132 to departing the of the the 7 has invention alternatives, and and of ordinary effects methods was been the 25 from mesenteric seen invention days, described observed to skill the and the with modifications significantly scope in particular applications as administration the were in artery defined connection of relevant dose the forms of and (p<0.05) by invention described dependent. spontaneously arts equivalents the set of with that following 10 forth, inhibited certain or mg/kg herein other any but as 2014202047 11 Apr 2014 perfluorobenzene. triflurobenzene, ketamine. methadone, phenmetrazine, phenylisopropylamine xylene, hallucinogenic 4. 2. of 9. 7. fenfluramine. 6. from 5. anxiolytic, agent, 3. abuse comprising 8. 1 CLAIMS . abuse amphetamine, and an A The The The The The The The The m-xylene, of, opioid, method a and administering method method method method method method method method hydrocodone, medication. dependence mushroom, an methylphenidate, 1,2,4-trifluorobenzene, a of illicit p-xylene, hallucinogen, of of of of of of of of dextroamphetamine, treating claim claim claim claim claim claim claim claim derivative. substance. on, to and lysergic or ethylbenzene, the 4, 2, 3, 3, 3, 3, a 3, 1, patient oxycodone. withdrawal wherein wherein wherein wherein wherein wherein wherein wherein patient an diethylpropion, acid inhalant, suffering a pentafluorotoluene, the the the the the the the the diethylamide and therapeutically fluorobenzene, from 133 psychostimulant psychostimulant beta-phenylisopropylamine hallucinogen inhalant opioid drug at a methamphetamine. least sedative, from at of pemoline, is least one abuse is selected or (LSD), selected substance susceptible one a effective is is tranquilizer, o-difluorobenzene, selected pentafluorobenzene, mazindol, selected substance, agent agent phencyclidine from from is amount is is Lortab, to from selected benzene, from a selected at beta (-) a the derivative least hypnotic, psilocybin, cathione, a of method Tramadol, psychostimulant (PCP), from Compound one from 1,3,5- toluene, and symptom is a an ecstasy, and and drug selected a heroin, o- A. of 2014202047 11 Apr 2014 blocker, monoamine norepinephrine reuptake therapeutically reduces nosebleeds; vertigo; reduces inhibitor, increases Lorcet, opium, inflammatory atypical antagonist, antagonist, (NDRI), extreme depression, arrhythmia; decreased sociability; attention analgesic, 14. 13. 12. 11 10. 15. . The The The The The The cannabinoid, Percocet, at at dizziness; antidepressant/antipsychotic, suspicion, a a inhibitor deficit at an calcium sensation an least least serotonin aNK-1 a less method method irritability, method method method method least nasal cardiomyopathy; oxidase adenosine central anticholinergic medication, effective reuptake one one hunger hyperactivity one Percodan, stuffiness; (SSRI), anxiety; channel and receptor of of of of of of symptom symptom of inhibitor alpha 5 cannabis, negative -hydroxytryptamine claim claim claim claim claim claim pain; sleep receptor craving and amount inhibitor an a adrenergic blocker, pychosis; fatigue; and serotonin-norepinephrine antagonist, bronchitis; facial 2, 2, disorders, agent, 1, 11, 11, 11, over heart (MAOI), of of disorder; subjective benzodiazapine wherein wherein wherein for antagonist, Tylox. wherein wherein wherein cocaine cocaine of (NRI), the pain; cocaine. attack; least an a a confusion; marked agonist, central a counter antihistamine, increased a euphoria; tricyclic, a shortness the the the dysphoria; one withdrawal abuse monoamine the the the a symptom corticotropin 1 dilated A norepinephrine-dopamine 134 medication an method drug drug other drug drug and (5HT1A) feeling medication, a adenosine carisprodol, and central nasal of an peripheral appetite, increased of pupils; of of of agent of dependence abuse and further anticonvulsant, breath; selected abuse abuse abuse of a oxidase cocaine reuptake irritation; releasing muscle or is antagonist, physical craving selected A2A nausea; selected is peripheral pyschomotor and is is is comprises tobacco, energy, chest alpha cocaine, from cocaine cocaine cocaine B abuse receptor relaxant, an inhibitor in inhibitor, for nasal factor and from vomiting; pain; antidepressant adrenergic the from fatigue, a cocaine. excitement a nicotine, and dopamine beta alcohol, mental reuptake glutamate and and and patient co-administering crusting; a (CRF) heart antagonist, an a retardation, (SNRI), selective dependence. adrenergic nonsteroidal the the the a anesthetic, lack sodium headache; strength; palpitations; receptor selected Compound Compound Compound caffeine, Vicodin, antagonist, inhibitor β and recurrent of antagonist, -hydroxylase medication. a serotonin pleasure, a channel agitation, receptor an from anti a A A A an a 2014202047 11 Apr 2014 phenylisopropylamine patient therapeutically 24. 23. 22. in 21 20. hydroxybutyrate, inhalant. opioid, anxiolytic, agent, abuse extinction acquisition from substance levodopa, direct a synthesis gamma-aminobutyric 19. 18. 17. 16. catechol-O-methyl-transferase the . acquisition, patient. or and an a A The The The The The The The The a indirect therapeutically carboxylase opioid, activator, method in carisprodol, a phase and phase method method method method method method method method medication. a patient, an effective in a dopamine maintenance, opium, in of illicit hallucinogen, the a of of of of of of of of the treating partial inhibitor, acid wherein modafenil, claim claim claim claim claim claim claim claim derivative. patient. patient. substance. effective amount psilopcybin, (GABA) agonist, dopamine 23, 22, at 18, 18, 18, 18, 1, 1, the wherein least wherein extinction, a wherein wherein wherein wherein inhibitor, wherein wherein of an partial amount acamprosate, at least inhalant, a agonist, one least partial hallucinogenic D2 the the opioid phase one the the the the the the of an one agonist, and 135 method Compound a Compound 5HT1 psychostimulant other drug at Compound Compound Compound a opioid GABA phase relapse, of sedative, agonist, least gamma-butyrolactone, substance of agent a agonist, further receptor one dopamine abuse of mushroom, metabolism comprising A substance a A. a A A A selected substance partial is tranquilizer, reduces promotes inhibits is comprises a dependence administered antagonist, serotonin selected agent metabolism nicotinic tobacco, from dependence enzyme administering is the the is the selected co-administering from a benzodiazepine, development a gamma frequency 5HT2 beta on hypnotic, development a to agonist, and mood inhibitor, enzyme at a the psychostimulant antagonist, least from is nicotine. patient selected stabilizer, to of and an one a inhibitor, the relapse of a drug of GABA an the after a the an of a a 2014202047 11 Apr 2014 blocker, monoamine norepinephrine reuptake therapeutically perfluorobenzene. triflurobenzene, ketamine. phenmetrazine, inhibitor, Percocet, inflammatory xylene, 29. hallucinogenic 28. heroin, 27. 26. 25. antagonist, antagonist, (NDRI), 32. cannabinoid, 3 analgesic, 30. fenfluramine. selected f . The The The The The The The methadone, m-xylene, The from a a inhibitor Percodan, an calcium an serotonin aNK-1 a method method method method method method method oxidase method adenosine central cannabis, anticholinergic medication, amphetamine, mushroom, effective reuptake methylphenidate, 1,2,4-trifluorobenzene, p-xylene, (SSRI), channel receptor of hydrocodone, of of of of of of and inhibitor of alpha 5 -hydroxytryptamine claim claim claim claim claim claim claim benzodiazapine, receptor claim Tylox. amount inhibitor an a adrenergic blocker, lysergic antagonist, serotonin-norepinephrine ethylbenzene, 18, dextroamphetamine, 22, 23, 23, 23, 23, 24, agent, over 22, (MAOI), antagonist, wherein wherein wherein wherein wherein wherein wherein of wherein diethylpropion, and (NRI), the least acid an a agonist, central oxycodone. counter antihistamine, a a pentafluorotoluene, carisprodol, diethylamide one the monoamine the the the the the the a corticotropin the 1 fluorobenzene, A norepinephrine-dopamine 136 an method psychostimulant opioid beta-phenylisopropylamine medication inhalant hallucinogen other and (5HT1A) drug a medication, adenosine central peripheral pemoline, and agent of is tobacco, further a oxidase abuse is methamphetamine. reuptake (LSD), selected releasing muscle or antagonist, selected is selected A2A peripheral o-difluorobenzene, is and selected is pentafluorobenzene, mazindol, alpha comprises selected nicotine, phencyclidine B agent alcohol, receptor relaxant, an from inhibitor inhibitor, from factor from antidepressant adrenergic a from is Lortab, dopamine beta reuptake from benzene, selected caffeine, (-) Vicodin, a co-administering (CRF) antagonist, a (SNRI), selective derivative an adrenergic nonsteroidal cathione, a psilocybin, sodium anesthetic, Tramadol, (PCP), receptor antagonist, 1,3,5- inhibitor from β toluene, opium, Lorcet, -hydroxylase medication. and a serotonin a is and channel and ecstasy, receptor a anti an o- a an 2014202047 11 Apr 2014 taken use major the occupational or it in inhalant. opioid, activities cocaine 39. exacerbated cocaine 38. symptom 37. 36. extinction 35. acquisition 34. comprising 33. direct a synthesis gamma-aminobutyric atypical catechol-O-methyl-transferase is the unsuccessful at despite physically in least patient. or role A The The The The The The a dependence abuse antidepressant/antipsychotic, larger indirect carboxylase to activator, method of phase obligations one phase having administering method method method method method method obtain by abuse or in amounts hazardous; phase the recreational efforts in the dopamine in of of, the persistent effects the a of of of of of of the in treating patient partial is inhibitor, acid dependence at cocaine, claim claim claim claim claim claim patient. to the patient. or selected work, cut to of recurrent over patient (GABA) agonist, selected dopamine activities the 37, 33, 33, 33, 33, at 37, the or down least school, use a recurrent a patient wherein wherein wherein wherein wherein wherein inhibitor, from cocaine. partial longer selected on, the or a cocaine-related agonist, one from partial a are acquisition, control or D2 or cocaine, tricyclic, a opioid period phase withdrawal the the the the the the therapeutically social given home; recurrent an agonist, from 137 a 5HT1 Compound Compound Compound Compound Compound Compound opioid cocaine GABA of agonist, then up or or tolerance; recurrent an cocaine maintenance, interpersonal recover or a agonist, cocaine legal anticonvulsant, receptor was dopamine from reduced metabolism use; a A A A A A A intended; effective problems; partial dependence cocaine cocaine. withdrawal; from a promotes reduces reduces reduces reduces inhibits a use great antagonist, serotonin because metabolism resulting its problems extinction, nicotinic deal enzyme amount use there the effects; the in at at a and development glutamate least least the in in of development of frequency the 5HT2 continued is a a situations in cocaine patient time agonist, caused of mood patient, inhibitor, a one one cocaine important enzyme and a persistent Compound failure antagonist, is symptom symptom relapse, antagonist, stabilizer, spent at use; or of and of wherein cocaine is in least inhibitor, relapse of a to the often social, which GABA and an desire in the fulfill one A. of of an the a a 2014202047 11 Apr 2014 the remission. pupils; breath; psychological 48. 47. 46. 45. WAIS-R, 44. Inventory, 43. 42. 41 irritation; 40. early and at craving disorders, cocaine subjective craving feeling euphoria; cocaine cocaine least . patient. frequency full nausea; of The The The The The The The The The chest one for withdrawal for use abuse remission, nasal increased WMS-R, physical increased symptom and cocaine. cocaine. of method method method method method method method method method is pain; problem continued ADHD-IV, and of a vomiting; crusting; cognitive cocaine dependence heart and RAVLT, of of of of of of of of of selected energy, appetite, of early claim claim claim claim claim claim claim claim claim cocaine that mental palpitations; despite recurrent use headache; partial HAM-D, function is excitement from 43, 46, 33, 33, 37, 37, 37, 33, 33, Trials pyschomotor by likely strength; selected abuse knowledge wherein wherein wherein wherein wherein wherein wherein wherein wherein the remission, fatigue, nosebleeds; rating I-VII, HAM-A, to vertigo; patient. arrhythmia; and have and from decreased the the the the the the the the the dependence. RCFT, scale. lack retardation, 138 sociability; been of sustained dizziness; remission Compound Compound Compound Compound Compound Compound cognitive Compound attention BDI, having nasal of caused and cardiomyopathy; pleasure, sensation apathy stuffiness; TMT, full a deficit function anxiety; is agitation, less persistent A A A A A A A or characterized remission, scale reduces reduces prolongs promotes reduces increases improves depression, exacerbated hunger Parts of hyperactivity pain; facial pychosis; rating from extreme A or heart at at at and and a bronchitis; recurrent at remission a and Neuropsychiatric pain; least least least period scale score irritability, least by fatigue; by B. attack; sustained suspicion, confusion; at one one one the dysphoria; disorder; is one of least of selected physical cocaine. of symptom symptom in the remission a dilated shortness negative marked the the one sleep patient partial and nasal patient. amount of and from or of of of in on 2014202047 11 Apr 2014 blocker, monoamine norepinephrine reuptake therapeutically inhalant. opioid, inhibitor, one 49. direct a synthesis gamma-aminobutyric atypical antagonist, antagonist, (NDRI), 50. catechol-O-methyl-transferase of or contingency The The a antidepressant/antipsychotic, a indirect a inhibitor carboxylase activator, an calcium serotonin aNK-1 a method method oxidase adenosine central effective reuptake Patent dopamine (SSRI), channel receptor management a of of inhibitor alpha 5 partial inhibitor, -hydroxytryptamine acid claim claim receptor Attorneys amount inhibitor a adrenergic (GABA) blocker, agonist, antagonist, serotonin-norepinephrine dopamine 33, 33, (MAOI), a antagonist, SPRUSON wherein inhibitor, wherein of and partial (NRI), Biotie least for a a agonist, cognitive partial agonist, central a D2 a the tricyclic, a opioid one monoamine the the a Therapies, corticotropin 1 an agonist, A norepinephrine-dopamine 139 Applicant/Nominated an & a 5HT1 method method other and opioid (5HT1A) a GABA FERGUSON adenosine behavioral agonist, central an peripheral agent a agonist, anticonvulsant, receptor dopamine further further Inc. oxidase metabolism reuptake releasing or antagonist, a selected A2A partial peripheral therapy. a alpha comprises comprises antagonist, serotonin B receptor metabolism inhibitor inhibitor, nicotinic factor from adrenergic enzyme a Person a dopamine beta reuptake glutamate a (CRF) treatment co-administering 5HT2 antagonist, (SNRI), selective a adrenergic a agonist, mood inhibitor, enzyme sodium receptor antagonist, antagonist, inhibitor β antagonist, -hydroxylase stabilizer, a with serotonin and a channel inhibitor, a receptor GABA at an least a an an a a 2014202047 11 Apr 2014 Figure 1/11 1 2014 2/11 Apr 11 Figure 2 ENZYME ASSAY Tyrosine hydroxylase Release of [3H2O] associated with conversion of L-[3,5-3H]-tyrosine to DOPA Acetyl CoA synthetase Utilization of [3H]-acetic sodium acetate 2014202047 Acyl-CoA, Cholesterol acyltransferase Formation of cholesteryl ester from [l-14C]-palmitoyl-CoA and endogenous cholesterol Ca2+/Calmodulin-protein kinase II Phosphorylation [32P] of BB40 (a synthetic peptide substrate) Cyclooxygenase-I Oxidation of arachidonic acid followed by spectrophotometric quantitaion of malondialdehyde HMG-CoA reductase Formation of [14C]-mevalonic acid fom [14C]-HMG-CoA Neutral endopeptidase (human) Formation of 4-methoxy-2-napthylamine from glutaryl-ala-ala-phe-4-methoxy-2- naphthylamide Nitric oxide synthase (constitutive) Conversion of [3H]-arginine to [3H]- citrulline Nitric oxide synthase (inducible) Measurement of iNOS reaction products (NO2 and NO3) in cytosol preparation from mouse macrophages induced by interferon-γ and lipopolysaccharide Phosphodiesterase III (human) Conversion of [3H]-cAMP to [3H]-AMP which is subsequently converted to [3H]~ adenosine Phospholipase Aj Formation of [l4C]-palmitate from [l4C]- 3- phosphatidylcholine Protein kinase C (non-selective) Phosphorylation [32P]of Histone Hl 2014202047 11 Apr 2014 Cyclooxygenase-I Ca Acyl-CpA: Acetyl Dopamine-B-hydroxylase Enzymes ENZYME Nitric Nitric HMG-CoA Dopamine-B-hydroxylase α Receptors Protein Phosphodiesterase Neutral 5-HT μ D Bi, a Phospholipase Mi -opioid 2A ι t a 2+ , B , D , muscarinic /Calmodulin 2 ccIB (X2B ia 2 oxide oxide adrenoceptors CoA dopamine Kinase , Endopeptidase 5-ΗΤ receptors adrenoceptors adrenoceptors OR Cholesterol Reductase Synthetase synthase synthase 2 RECEPTOR A (non-selective) α receptors , 2 Protein receptors 5-HT ΙΠ (inducible) (constitutive) (human) (human) Acyltransferase 2 c Kinase (human) (bovine) serotonin Π receptors Figure 3/11 3 9nM IC50 < < < < < < < 8.5 < < < <5 <5 <5 <5 < <5 <5 <5 10 10 10 10 10μΜ 10 10 10 10 10 ΙθμΜ nM (enzyme) μΜ μΜ μΜ μΜ μΜ μΜ μΜ μΜ μΜ or pKi (receptor) 2014202047 11 Apr 2014 £ Tt 6J0 S Z ^■4 U CONCENTRATION OF % INHIBITION x ENZYME NEPICASTAT (μΜ) OF ENZYME ' CM - 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