Note: Descriptions are shown in the official language in which they were submitted.
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W O 97/41879 PCTrUS97/07516
TITLE OF THE INVENTION
ENHANCEMENT OF SLEEP WITH A GROWTH HORMONE
SECRETAGOGUE
5 BACKGROUND OF THE INVENTION
Although sleep is necessary for survival, its precise
homeostatic contribution is unknown. Sleep is not a uniform state, but
rather involves several stages characterized by changes in the individual's
EEG. A non rapid eye movement (NREM) type (75 to 80% of total sleep
10 time) ranges in depth through stages 1 to 4 (deepest level). Stage 1 sleep
is drowsiness, in which the EEG displays a lower voltage, more mixed
frequencies and deterioration of alpha rhythm relative to the EEG when
the individual is awake. In stage 2, background activity ~imil~r to that of
stage 1 is experienced, with bursts of slightly higher ~requency "sleep
15 spindles" and sporadic higher amplitude slow wave complexes. The third
and fourth stages of sleep display increasing high amplitude slow wave
activity. The separate sleep stage in which the individual undergoes rapid
eye movement (REM) occupies the remainder of the sleep time and
occurs 5 to 6 times during a normal nights sleep. REM sleep is
20 characterized by a lower voltage, higher frequency EEG and other
characteristics simil~r to those which occur when the individual is awake,
whereas the other four sleep stages are categorized as NREM sleep.
Individuals vary widely in their requirements for sleep,
which is influenced by a number of factors including their current
25 emotional state. The natural aging process is associated with changes in a
variety of circadian and diurnal rhythms. Age-related changes in the
timing and structure of sleep are surprisingly common problems for older
people, and are often associated with significant morbidity. With
advancing age, the total amount of sleep tends to shorten. Stage 4 can
30 decrease or disappear and sleep may become more fragmented and
...... . ~ . .. ..
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W O 97/41879 PCTrUS97/07516
interrupted. Evaluation of sleep patterns in elderly people shows that the
timing of sleep is also phase advanced, especially in women. This
tendency to go to sleep and wake up earlier is very frustrating to older
people who feel that they are out of step with the rest of the world. In
S addition, the quality of sleep in the elderly is ~limini~hed with a m~rkçtlreduction in slow wave sleep, a reduction in the deep stages of sleep
(especially stage 4), fragmentation of REM sleep and more frequent
awakenings. Similarly, non-elderly people may exhibit disturbances in
the normal sleep process. These changes in the structure of sleep have
been correlated to more frequent napping, decreased daytime alertness
and declining intellectual function and cognitive ability. Deprivation of
REM sleep has been suggested to interfere with the memory
consolidation involved in learning skills through repetitive activity, and
slow wave sleep has been implicated as being important in consolidation
of events into long term memory. Likewise, decreases in the length of
REM stages of sleep may be associated with a decrease in cognitive
function and learning, especially ~limini~hed retention of memory.
Sleep disorders generally involve disturbances of sleep that
affect a subject's ability to fall and/or stay asleep, and involve sleeping
too little, too much or resulting in abnormal behavior associated with
sleep.
Numerous compounds are employed in the art to facilitate
normal sleep and to treat sleep disorders and sleep disturbances, including
e.g., sedatives, hypnotics, anxiolytics, antipsychotics, ~nti~nxiety agents,
minor tranquilizers, melatonin agonists and antagonists, melatonergic
agents, benzodiazepines, barbituates, SHT-2 antagonists, and the like.
Similarly, physical methods have been employed to treat patients with
sleep disorders such as the use of light therapy or the application of
morl~ te~l electrical .~i~n~l~ to selected nerves or nerve bundles.
Nevertheless7 the known threapeutic regimens suffer from
numerous problems, including residual effects in daytime function,
impairment of memory, potential for addiction, rebound insomnia, "REM
rebound" which may be associated with increased dream intensity and the
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occurrence of ni~htm~res, and the like. Accordingly, a more
physiological way to enhance sleep would be highly desirable.
Growth hormone, which is secreted from the pituitary,
stimulates growth of all tissues of the body that are capable of growing.
5 In addition, growth hormone is known to have the following basic effects
on the metabolic processes of the body: (1) Increased rate of protein
synthesis in all cells of the body; (2) Decreased rate of carbohydrate
tili7~tion in cells of the body; (3) Increased mobilization of free fatty
acids and use of fatty acids for energy.
A deficiency in growth hormone secretion can result in
various medical disorders, depending on the age of onset. In children, the
syndrome is characterized by short stature with normal body proportions
and reduced growth rate (dwarfism). A deficiency in growth hormone
secretion in adult life may be characterized by excessive adiposity,
reduced muscle mass, impaired exercise capacity, reduced body water,
decreased bone mineral density, and psychological disorders.
The temporal association of growth hormone secretion and
sleep is well established. In particular, growth hormone release and slow
wave sleep both predomin~te in the first part of sleep. Maximal growth
hormone secretion is observed during stages 3 and 4 (slow wave) sleep.
Additional studies, however, suggest that although slow wave sleep and
growth hormone secretion occur simultaneously (and may influence each
other), there may be another stimulus or series of neural events which
trigger both processes.
A study of the interaction between sleep and growth
hormone indicates that subjects with growth hormone disturbances have
abnormal REM and delta sleep and that norm~li7~tion of growth hormone
levels corrects the sleep stages (Astrom, Acta. Neurol. Scand., 2~, 281-
286 (1995); Astrom, etal., Neuroendocrinology~ 51, 82-84 (1990)). In
particular, young adults with lack of growth hormone compared to
normal subjects revealed increased total sleep time, decreased delta sleep
time, normal total REM sleep time, and increased stage 1 and stage 2
sleep time. Young adults with high growth hormone concentration
(acromegalic) compared to normal subjects revealed decresed REM sleep
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W O 97/41879 PCT~US97/07516
time, decresed delta sleep time, and increased stage 2 and stage 1 sleep
time. A~lmini~tration of systemic growth hormone, however, is reported
to not affect human sleep (Kern, et al., J. Clin. Endo. Metab., 76, 1428-
1432 (1993)). Gamma-hydroxybutyrate has been reported to be useful
5 for increasing slow-wave sleep in patients exhibiting low levels of slow
wave sleep (see e.g. PCT Patent Pub. No. WO 96/40105). Effects of oral
~1mini.~tration of a growth hormone secretagogue to healthy young men
(Copinschi, et al., J. Clin. Endo. Metab.~ 81 (8), 2776-2782) and to healthy
elderly subjects have been reported (Chapman, et al., J. Clin. Endo.
10 Metab81(12) 4249-4257 (1996).
Various ways are known to release growth hormone. For
example, chemicals such as arginine, L-3,4-dihydroxyphenyl~l~nine
(L-DOPA), glucagon, vasopressin, and insulin induced hypoglycemia, as
well as activities such as sleep and exercise, indirectly cause growth
hormone to be released from the pituitary by acting in some fashion on
the hypoth~l~mus perhaps either to decrease somatostatin secretion or to
increase the secretion of the known growth hormone secretagogue growth
hormone releasing factor (GRF) or an unknown endogenous growth
hormone-releasing hormone or all of these.
In cases where increased levels of growth hormone were
desired, the problem was generally solved by providing exogenous
growth hormone or by ~(lmini~tering GRF, IGF-I or a peptidal compound
which stimulated growth hormone production and/or release. In either
case the peptidyl nature of the compound necessitated that it be
~ ini~tered by injection. ~niti~lly the source of growth hormone was the
extraction of the pituitary glands of cadavers. This resulted in a very
expensive product and carried with it the risk that a disease associated
with the source of the pituitary gland could be transmitted to the recipient
of the growth horrnone. Recombinant growth hormone has become
available which, while no longer carrying any risk of disease
tr~n~mi.~sion, is still a very expensive product which must be given by
injection or by a nasal spray. In addition, ~-1mini.~tration of exogenous
growth hormone may result in side-effects, including edema, and does not
CA 022~3390 1998-10-29
W O97141879 PCTrUS97/07516 -
co~elate with the pulsitile release seen in the endogenous release of
growth hormone.
Certain compounds have been developed which stimulate the
release of endogenous growth hormone. Peptides which are known to
S stimulate the release of endogenous growth hormone include growth
hormone releasing hormone, the growth ho~none releasing peptides
GHRP-6 and GHRP-l (described in U.S. Patent No. 4,411,890, PCT
Patent Pub. No. WO 89/07110, and PCT Patent Pub. No. WO 89/07111)
and GHRP-2 (described in PCT Patent Pub. No. WO 93/04081), as well
as hexarelin (J. Endocrinol Invest., lS(Suppl 4), 45 (1992)).
Growth holmone releasing peptide-6 (GHRP-6) is believed
to act via a unique pituitary and hypoth~l~mic receptor and stim~ tes the
release of growth ho~none, corhsol and prolactin. In a study in young
men, repeated intravenous boluses of GHRP-6 increased stage 2 sleep
without affecting slow wave or REM sleep (Frieboes, et al.,
Neuroendocrinolo~y, 61:584-589 (l99S)). Growth honnone-releasing
hormone is reported to mediate feeding-specific feedback to the
suprachi~m~tic circadian clock (Vaccarino, et al., Peptides~ 16(4), 595-
598 (l99S). Although the effects of systemic ~lmini.~tration of growth
hormone-releasing honnone on sleep are not consistent, episodic
~rlmini.~tration of growth horrnone-releasing hormone is reported to
promote slow wave sleep and enhance REM sleep, relative to placebo or
continuous infusion of growth hormone-releasing hoImone (Marshall, et
al., J. Clin. Endocrinol. Metab., 81(3), 1009-1013 (1996)). Nevertheless,
a disadvantage of these peptidal growth hormone secretagogues is that
they have veIy low oral bioavailability. Although oral ~dmini~tration of
GHRP-2 is under clinical investigation, all of the other peptidal growth
holmone secretagogues must be ~1mini.etered parenterally.
Other compounds possessing growth hormone secretagogue
~ 30 activity are disclosed in the following: U.S. Patent No. 3,239,345; U.S.
Patent No. 4,036,979; U.S. Patent No. 4,411,890; U.S. Patent No.
S,206,235; U.S. Patent No. 5,283,241; U.S. Patent No. 5,284,841; U.S.
Patent No. 5,310,737; U.S. Patent No. 5,317,017; U.S. Patent No.
5,374,721; U.S. Patent No. 5,430,144; U.S. Patent No. 5,434,261; U.S.
,.. ,. ., . .. . ..... ~.
CA 022~3390 1998-10-29
WO 97/41879 PCT/US97/07516
Patent No. 5,438,136; U.S. Patent No. 5,494,919; U.S. Patent No.
5,494,920; U.S. Patent No. 5,492,916; U.S. Patent No. 5,536,716; EPO
Patent Pub. No. 0,144,230; EPO Patent Pub.No. 0,513,974; PCT Patent
Pub.~o. W 0 94/07486; PCT Patent Pub.No. W 0 94/08583; PCT Patent
Pub.No. W 0 94/11012; PCT Patent Pub.No. W 0 94/13696; PCT Patent
Pub. No. WO 94/19367; PCT Patent Pub. No. WO 95/03289; PCT Patent
Pub. No. WO 95/03290; PCT Patent Pub. No. WO 95/09633; PCT Patent
Pub. No. WO 95/11029; PCT Patent Pub. No. WO 95/12598; PCT Patent
Pub. No. WO 95/13069; PCT Patent Pub.No. W 0 95/14666; PCT Patent
Pub. No. WO 95/16675; PCT Patent Pub. No. WO 95/16692; PCT Patent
Pub. No. WO 95/17422; PCT Patent Pub. No. WO 95/17423; PCT Patent
Pub. No. WO 95/34311; PCT Patent Pub. No. WO 96/02530; PCT Patent
Pub. No. W 0 96/05195; PCT Patent Pub. No. WO 96/15148; PCT Patent
Pub. No. WO 96/22782; PCT Patent Pub. No. WO 96/22997; PCT Patent
Pub. No. WO 96/24580; PCT Patent Pub. No. WO 96/24587; PCT Patent
Pub. No. WO 96/35713; PCT Patent Pub. No. WO 96/38471; PCT Patent
Pub.No. W 0 97/00894; PCT Patent Pub. No. WO 97/06803; PCT Patent
Pub. No. WO 97/07117; Science. ~, 1640-1643 (June 11, 1993); Ann.
Rep. Med. Chem., 28, 177-186 (1993); Bioorg. Med. Chem. Ltrs., 4(22),
2709-2714 (1994); and Proc. Natl. Acad. Sci. USA 92, 7001-7005 (July
1995). Additional compounds with growth hormone secretagogue
activity are described herein.
SUMMARY OF THE INVENTION
The present invention is directed to the use of a compound
which has the ability to stimulate or amplify the release of natural or
endogenous y,rowlh hormone or growth horrnone-releasing hormone for
enhancing or improving sleep quality, in particular by increasing sleep
efficiency and augmenting sleep maintenance, as well as for preventing
and treating sleep disorders and sleep disturbances, in a warm-blooded
~nim~l. The known threapeutic regimens regarding sleep suffer from
numerous problems, including residual effects in daytime function,
impairment of memory, potential for addiction, rebound insomnia, "REM
rebound" which may be associated with increased dream intensity and the
CA 022~3390 1998-10-29
WO 97/41879 PCT/US97/07516
occurrence of ni~htm~res, and the like. An advantage of the present
method is that it provides a physiological-like pulsatile profile of growth
hormone release from the pituitary gland and further provides for the
release of growth hormone-releasing hormone. Accordingly, the present
5 invention provides a method for enhancing or improving sleep quality
and increasing sleep efficiency and sleep maintenance in a warm-blooded
Anim~l comprising the ~mini.~tration of a growth hormone secretagogue.
The present invention further provides a pharmaceutical composition for
enhancing or improving sleep quality and increasing sleep efficiency and
10 sleep maintenance.
DESCRIPTION OF THE INVENTION
The present invention is directed to the use of a compound
which has the ability to stimulate or amplify the release of natural or
15 endogenous growth hormone for enhancing or improving sleep quality as
well as preventing and treating sleep disorders and sleep disturbances in a
warm-blooded :l~im~l. In particular, the present invention provides a
method for enhancing or improving sleep quality by increasing sleep
efficiency and augmenting sleep maintenance. In addition~ the present
20 invention provides a method for preventing and treating sleep disorders
and sleep disturbances in a warm-blooded ~nim~l which comprising the
~tlmini~tration of a growth hormone secretagogue. The present invention
further provides a pharmaceutical composition for enhancing or
improving sleep quality and increasing sleep efficiency and sleep
25 maintenance
The following outcomes in a subject which are provided by
the present invention may be correlated to enhancement in sleep ~uality:
an increase in the value which is calculated from the time that a subject
sleeps divided by the time that a subject is attempting to sleep; a decrease
30 in sleep latency (the time it takes to fall asleep); a decrease in difficulties
in falling asleap; a decrease in the number of awakenings during sleep; a
decrease in nocturnal arousals; a decrease in the time spent awake
following the initial onset of sleep; an increase in the total amount of
sleep; an increase the amount and percentage of REM sleep; an increase
.. ... . , .. , ~ , .. , j,
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in the duration and occurrence of REM sleep; a reduction in the
fragmentation of REM sleep; an increase in the amount and percentage of
slow-wave (i.e. stage 3 or 4) sleep; an increase in the amount and
percentage of stage 2 sleep; an enhancement of EEG-delta activity during
5 sleep; a decrease in the number of awakenings; a decrease in nocturnal
arousals, especially early morning awakenings; an increase in daytime
alertness; an increased satisfaction with the intensity of sleep; and
increased sleep m~intenance. Secondary outcomes which may be
provided by the present invention include enhanced cognitive function
10 and increased memory retention.
The present invention is further useful for the prevention and
treatment of sleep disorders and sleep disturbances including sleep
problems associated with insomnia, hypersomnia, sleep apnea,
narcolepsy, nocturnal myoclonus, REM sleep interruptions, jet-lag, shift
15 workers' sleep disturbances, dysomnias, night terror, insomnias associated
with depression or with emotional/mood disorders, as well as sleep
walking and enuresis, as well as sleep disorders which accompany aging.
Sleep disorders and sleep disturbances are generally characterized by
difficulty in initiating or m~int~ining sleep or in obt~inin~ restful or
20 enough sleep. Simil~rly~ the present invention is useful for treating
conditions associated with circadian rhythmicity as well as mental and
physical disorders associated with travel across time zones and with
rotating shift-work schedules. In addition, certain drugs may also cause
reductions in REM sleep as a side effect and the present invention may be
25 used to correct those types of sleeping disorders as well. The present
invention would also be of benefit in the treatment of syndromes such as
fibromyalgia which are manifested by non-restorative sleep and muscle
pain or sleep apnea which is associated with respiratory disturbances
during sleep. In this case, the release of growth hormone associated with
30 the growth hormone secretagogue may also improve respiratory function
as a result of increased muscle strength and tone associated with growth
hormone release. It will be clear to one skilled in the art that the present
invention is not limited to just sleep disorders and sleep disturbances, but
CA 02253390 1998-10-29
WO 97/41879 PCT/US97/07516
is applicable to a wide variety of conditions which result from a
dimini.~hed quality of sleep.
In the present invention, it is preferred that the subject
nl~mm~l iS a human. Although the present invention is applicable both
old and young people, it would find greater application in elderly people.
Further, although the invention may be employed to enhance the sleep of
healthy people, it may be especially beneficial for enhancing the sleep
quality of people suffering from sleep disorders or sleep disturbances.
By the term "growth hormone secretagogue" is meant any
exogenously ~dminictered compound or agent that directly or
indirectly stimulates or increases the endogenous release of growth
hormone, growth hormone-releasing hormone or somatostatin in an
~nim~l, in particular, a human.
The growth hormone secretagogue may be peptidal or
non-peptidal in nature, however, the use of a non-peptidal growth
hormone secretagogue is preferred. In addition, for convenience the
use of an orally active growth hormone secretagogue is preferred. In
addition, it is preferred that the growth hormone secretagogue induce
or amplify a pulsatile release of endogenous growth hormone. It is
also preferred that the growth hormone secretagogue be able to cause
the release of growth hormone at night or during the sleep cycle,
especially in the first half of the night or of the sleep cycle, and even
more especially in the first few hours following sleep onset, or
alternatively in the period immediately preceding sleep onset.
The growth hormone secretagogue may be used alone or in
combination with other growth hormone secretagogues or with other
agents which are known to be beneficial in the enhancement of sleep
efficiency. The growth hormone secretagogue and the other agent may
be co~lmini~tered, either in concomitant therapy or in a fixed
combination. For example, the growth hormone secretagogue may be
~dmini~tered in conjunction with other compounds which are known in
the art to be useful for enhancing sleep quality and preventing and
treating sleep disorders and sleep disturbances, including e.g., sedatives,
hypnotics, anxiolytics, antipsychotics, ~nti~nxiety agents, minor
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W O97/41879 PCTrUS97/07516 -
- 10-
tranquilizers, melatonin agonists and antagonists, melatonergic agents,
benzodiazepines, barbituates, SHT-2 antagonists, and the like, such as:
~llin~7.olarn, allobarbital, alonimid, alprazolam, amitriptyline,
amobarbital, amoxapine, bentazepam, benzoct~mine, brotizolam,
S bupropion, busprione, butabarbital, butalbital, capuride, carbocloral,
chloral betaine, chloral hydrate, chlordiazepoxide, clomi~ e,
cloperidone, clorazepate, clorethate, clozapine, cyprazepam, desipramine,
dexclamol, diazepam, dichloralphenazone, divalproex, diphenhydramine,
doxepin, estazolarn, ethchlorvynol, etomidate, fenobam, flunitrazepam,
10 flurazepam, fluvoxamine, fluoxetine, fosazepam, glutethimide,
halazepam, hydroxyzine, imipramine, lithium, lorazepam, lorrnetazepam,
maprotiline, mecloqualone, melatonin, mephobarbital, meprobamate,
methaqualone, midaflur, midazolam, nefazodone, nisobamate,
nitrazepam, nortriptyline, oxazepam, paraldehyde, paroxetine,
15 pentobarbital, perlapine, pelphenazine, phenelzine, phenobarbital,
prazepam, promethazine, propofol, protriptyline, quazepam, reclazepam,
roletamide, secobarbital, sertraline, suproclone, temazepam, thioridazine,
tracazolate, tranylcypromaine, trazodone, triazolam, trepipam,
tricetarnide, triclofos, trifluoperazine, trimetozine, trimipramine,
20 uldazepam, venlafaxine, zaleplon, zolazeparn, zolpidem, and salts
thereof, and combinations thereof, and the like, or the growth horrnone
secretagogue may be ~ ini~tered in conjunction with the use of physical
methods such as with light therapy or electrical stimulation.
Representative growth horrnone secretagogues are disclosed
25 in: U.S. Patent No. 3,239,345; U.S. Patent No. 4,036,979; U.S. Patent
No. 4,411,890; U.S. Patent No. S,206,235; U.S. Patent No. 5,283,241;
U.S. Patent No. 5,284,841; U.S. Patent No. 5,310,737; U.S. Patent No.
5,317,017; U.S. Patent No. 5,374,721; U.S. Patent No. 5,430,144; U.S.
Patent No. 5,434,261; U.S. Patent No. 5,438,136; U.S. Patent No.
30 5,494,919; U.S. Patent No. 5,494,920; U.S. Patent No. 5,492,916; U.S.
Patent No. 5,536,716; EPO Patent Pub. No. 0,144,230; EPO Patent Pub.
No. 0,513,974; PCT Patent Pub. No. W 0 89/07110; PCT Patent Pub.No.
WO 89/07111; PCT Patent Pub. No. WO 93/04081; PCT Patent Pub.No.
WO 94/07486; PCT Patent Pub. No. WO 94/08583; PCT Patent Pub. No.
CA 022~3390 1998-10-29
WO 97/41879 PCT/US97/07516
WO 94/11012; PCT Patent Pub. No. WO 94/13696; PCT Patent Pub. No.
WO 94/19367; PCT Patent Pub. No. W 0 95/03289; PCT Patent Pub. No.
WO 95/03290; PCT Patent Pub. No. W 0 95/09633; PCT Patent Pub. No.
WO 95/11029; PCT Patent Pub. No. W 0 95/12598; PCT Patent Pub. No.
5 WO 95/13069; PCT Patent Pub. No. WO 95/14666; PCT Patent Pub. No.
WO 95/16675; PCT Patent Pub. No. WO 95/16692; PCT Patent Pub. No.
WO 95/17422; PCT Patent Pub. No. WO 95/17423; PCT Patent Pub. No.
WO 95/34311; PCT Patent Pub. No. WO 96/02530; PCT Patent Pub. No.
WO 96/05195; PCT Patent Pub. No. W 0 96/15148; PCT Patent Pub. No.
10 WO 96/22782; PCT Patent Pub. No. WO 96/22997; PCT Patent Pub. No.
WO 96/24580; PCT Patent Pub. No. WO 96/24587; PCT Patent Pub. No.
WO 96/35713; PCT Patent Pub. No. WO 96/38471; PCT Patent Pub. No.
WO 97/00894; PCT Patent Pub. No. WO 97/06803; PCT Patent Pub. No.
WO 97/07117; J. Endocrinol Invest., 15(Suppl 4), 45 (1992)); Science~
15 ~Q 1640-1643 (June 11, 1993); Ann. Rep. Med. Chem., 28, 177-186
(1993); Bioor~. Med. Chem. Ltrs., 4(22), 2709-2714 (1994); and Proc.
Natl. Acad. Sci. USA 2~. 7001-7005 (July 1995).
A representative first class of growth hormone secretagogues
20 is set forth in U.S. Patent No. 5,206,235 as follows:
R~(X)n-(CH2)p /R4
[ JJ, ~X - C - A - N
(C ~2)q
( --)W
R1a
R2a R3a
wherein the various substituents are as defined in U.S. Patent 5,206,235.
The most preferred compounds within this first class are
identified as having the following structures:
CA 02253390 1998-10-29
WO 97/41879 PCT/US97/07516
CH3 /H3
N--C--CH2--C--NH2
N~ NH
or
O CH3 /H3 OH
~ N~j,NH
A representative second class of growth hormone
5 secretagogues is set forth in U.S. Patent No. 5,283,241 and PCT Patent
Publication No. 94/05634 as having the following structural formula:
Rj~/ (X)n-(c -12)p/ R4
R~ IN--~ I 6 1I R5
(cHlJ2)q
(L)w
F~2a~ 3a
CA 02253390 1998-10-29
WO 97J41879 PCT/US97/07516
- 13-
wherein the various substituents are as defined in U.S. Patent 5,283,241
and PCT Patent Publication No. 94/05634.
A representative third class of growth hormone
S secretagogues is disclosed in PCT Patent Pub. No. WO 94/13696 as
compounds of the following structural Formulas I and II:
~2 ~6 R4 ~2 ~6 R4
R1--C--N-C--A N ~ R5
C=O C=O
BS~F)n B~F)n
R
3b R
3b
Formula I Formula 11
wherein:
Rl is selected from the group consisting of:
10 -Cl-Clo aLkyl, -aryl, -aryl-(Cl-C6 alkyl),
-C3-C7 cycloaLkyl-(C 1 -C6alkyl), -C 1 -C5alkyl-K-C 1 -C5 alkyl, -aryl(Co-
CsaLkyl)-K-(Cl-Cs aLkyl),
-C3-C7 cycloaLkyl(Co-C5 aLkyl)-K-(Cl-C5 alkyl),
wherein K is O, S(O)m, N(R2)C(O), C(O)N(R2), OC(O), C(O)O, or
15 -CR2-CR2-, or-C_C-,
and wherein the aryl groups are as defined below and the R2 and alkyl
groups may be futher substituted by 1 to 9 halogen, S(O)mR2a, 1 to 3
OR2a, or C(O)OR2a, and the aryl groups may be further substituted by
phenyl, phenoxy, halophenyl, 1-3 Cl-C6 alkyl, 1 to 3 halogen, 1 to 2
20 -OR2, methylenedioxy, -S(O)mR2, 1 to 2-CF3, -OCF3, nitro,
CA 022~3390 1998-10-29
W O97/41879 PCTrUS97/07516 -
- 14 -
-N(R2)(R2),-N(R2)C(O)R2,-C(O)OR2,-C(O)N(R2)(R2),
-SO2N(R2)(R2), -N(R2)S(O)2 aryl, and-N(R2)SO2R2;
R2 is selected from the group consisting of:
hydrogen, Cl-C6 alkyl, C3-C7 cycloalkyl, and where two Cl-C6 alkyl
5 groups are present on one atom, they may be optionally joined to form a
C3-Cg cyclic ring optionally including oxygen, sulfur or NR2a;
R2a is hydrogen, or Cl-C6 alkyl;
10 R3a and R3b are independently selected from the group consisting of:
hydrogen, halogen, -Cl-C6 alkyl, -OR2, cyano, -OCF3, methylenedioxy,
nitro, -S(O)mR, -CF3 or -C(O)OR2 and when R3a and R3b are in an
ortho arrangement, they may be joined to form a C5 to C8 aliphatic or
aromatic ring optionally including 1 or 2 heteroatoms selected from
15 oxygen, sulfur or nitrogen;
R4 and R5 are independently selected from the group consisting of:
hydrogen, -Cl-C6 alkyl, substituted Cl-c6 alkyl wherein the substituents
are selected from 1 to 5 halo, 1 to 3 hydroxy, 1 to 3
20 Cl-Clo alkanoyloxy, 1 to 3 Cl-C6 alkoxy, phenyl, phenoxy, 2-furyl, Cl-
C6 alkoxycarbonyl, -S(O)m(Cl-C6 alkyl); or R4 and R5 can be taken
together to form -(cH2)rLa (CH2)s where La is -C(R2)2-, -O-, -S(O)m-,
or -N(R2)-, where r and s are independently 1 to 3 and R2 is as defined
above;
R6 is hydrogen or Cl-C6 alkyl;
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Als:
R7
(CH2)x--I (CH2)y--
R7a
or
R7
- Z-(CH2)X--I (CH2)y
R7a
wherein x and y are independently 0-3;
ZisN-R2orO;
5 R7 and R7a are independently selected from the group consisting of:
hydrogen, -Cl-C6 alkyl, -OR2, trifluoromethyl, phenyl, substituted
C1-C6 alkyl where the substituents are selected from imidazolyl, phenyl,
indolyl, p-hydroxyphenyl, -OR2, 1 to 3 fluoro, -S(O)mR2, -C(O)OR2,
-C3-C7 cycloalkyl, -N(R2)(R2)~ -C(O)N(R2)(R2); or R7 and R7a can
10 independently be joined to one or both of R4 and R5 groups to form
aL~ylene bridges between the terminal nitrogen and the alkyl portion of
the R7 or R7a groups, wherein the bridge contains 1 to 5 carbons atoms;
B, D, E, and F are independently selected from the group consisting of:
15 -C(Rg)(Rlo)-, -O-, C=O, -S(O)m-, or -NRg, such that one or two of B,
D, E, or F may be optionally absent to provide a 5, 6, or 7 membered
ring; and provided that B, D, E and F can be -C(Rg)(R1o)- or C=O only
when one of the rem~inin~: B, D, E and F groups is simultaneously -O-,
-S(O)m-, or-NRg, or
20 B and D, or D and E taken together may be -N-CR1o- or -CRlo=N-,
or B and D, or D and E taken together may be -CR8=CRlo-, provided
one of the other of B and E or F is simultaneously -O-, -S(O)m-, or
-NRg-;
........ . . .. . ... .
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- 16-
R8 and Rlo are independently selected from the group consisting of:
hydrogen, -R2, -OR2, (-cH2)q-aryL -(CH2)q-C(O)OR2, -(CH2)q-
C(o)o(cH2)q-aryL or -(cH2)q-(lH-tetrazol-s-yl)~ where the aryl may be
optionally substituted by 1 to 3 halo, 1 to 2 C l-Cg alkyl, 1 to 3 -OR2 or 1
5 to 2 -C(O)OR2;
Rg is selected from the group consisting of:
-R2, -(cH2)q-aryL -C(O)R2, -c(o)(cH2)q-aryL -S02R2,
-so2(cH2)q-aryl, -C(O)N(R2)(R2), -c(o)N(R2)(cH2)q-aryl~
10 -C(O)OR2, 1-H-tetrazol-5-yl,-S03H,-S02NHC-N,-S02N(R2)aryl,
-S02N(R2)(R2),
and wherein the (CH2)q may be optionally substituted by 1 to 2 C 1 -C4
aL~yl, and the R2 and aryl may be optionally further substituted by 1 to 3
-OR2a, -0(CH2)q aryl, 1 to 2 -C(O)OR2a, 1 to 2 -C(O)O(CH2)q aryl, 1
15 to 2 -C(O)N(R2a)(R2a), 1 to 2 -C(O)N(R2a)(CH2)q aryl, 1 to 5 halogen,
1 to 3 Cl-C4 aL~yl, 1,2,4-triazolyl, 1-H-tetrazol-5-yl, -C(O)NHS02R2a,
-S(O)mR2a, -c(o)NHso2(cH2)q-aryL -so2NHc-N~ -so2NHc(o)R
-so2NHc(o)(cH2)qaryL -N(R2)C(O)N(R2a)(R2a),
-N(R2a)c(o)N(R2a)(cH2)q-aryL -N(R2a)(R2a)~ -N(R2a)c(o)R2a~
20 -N(R2a)C(O)(CH2)q aryl, -oc(o)N(R2a)(R2a)~ -0C(~)N(R2a)(CH2)q
aryl, -S02(CH2)qCONH-(CH2)wNHc(O)Rl 1,
wherein w is 2-6 and Rl 1 may be biotin, aryl, or aryl substituted by 1 or 2
OR2, 1-2 halogen, azido or nitro;
25 misO, 1 or2;
n is 1, or 2;
q may optionally be 0, 1, 2, 3, or 4; and
G, H, I and J are carbon, nitrogen, sulfur or oxygen atoms, such that at
least one is a heteroatom and one of G, H, I or J may be optionally
30 missing to afford a 5 or 6 membered heterocyclic aromatic ring;
and pharmaceutically acceptable salts and individual diastereomers
thereof.
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WO 97/41879 PCT/US97/07516 --
Within this third class, the most preferred growth hormone
secretagogues employed in the instant invention are realized in structural
Formula V:
CH3
- N~NH2
~o o
D~R3a
S V
wherein R1 is selected from the group consisting of:
CH2CH2-, ~ CH2CH2CH2-~3CH2ocH2
~3~CH2 F~CH2- ~
~CH2CH2~ 'CH2CH2CH2-~ ~CH2CH2-.
F~ CH2CH2CH2-;
R3a is H, or fluoro;
D is is selected from the group consisting of:
-O-, -S-, -S(O)m-, N(R2)~ Nso2(R2)~ Nso2(cH2)taryL NC(~)(R2),
NSO2(CH2)qOH, NSO2(CH2)qCOOR2, Nso2(cH2)qc(o)-N(R2)(R2)~
N-S02(CH2~qC(O)-N(R2)(CH2~wOH,
.. . . ..
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- 18-
N-so2(cH2)qc(o)-N(R2)(cH2)w J~
HN~NH
o
o OH
N~SO2(CH2)qC(O)-N(R2)(cH2)w --N ~ N3
N-NH
N-SO2(CH2)q ~/
N=N
and the aryl is phenyl or pyridyl and the phenyl may be
S substituted by 1-2 halogen;
R2 is H, or Cl-C4 alkyl;
mis 1,2;
tisO, 1,or2;
10 qis 1,2,or3;
w is 2, 3, 4, 5, or 6;
and the ph~ ceutically acceptable salts and individual diastereomers
thereof.
Representative most preferred growth hormone secretagoues
within this third class which may be employed in the present invention
include the following:
1) N-[l(R)-[(1,2-Dihydro-l-methanesulfonylspiro[3H-indole-3,4'-
piperidin]- 1 '-yl)carbonyl]-2-( 1 H-indol-3-yl)ethyl]-2-amino-2-methyl-
propanamide;
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- 19-
2) N-[1 (R)-[( 1 ,2-Dihydro- l-methanecarbonylspiro[3H-indole-3,4'-
piperidin]- 1 '-yl)carbonyl]-2-( 1 H-indol-3-yl)ethyl] -2-amino-2-methyl-
prop~n7~mide;
5 3) N-[l(R)-[(1,2-Dihydro-1-benzenesulfonylspiro[3H-indole-3,4'-
piperidin]- 1 '-yl)carbonyl]-2-(lH-indol-3-yl)ethyl]-2-amino-2-methyl-
prop~n~mide;
4) N-[l(R)-[(3,4-Dihydro-spiro[2H-l-benzopyran-2,4'-piperidinJ-l'-yl)
10 carbonyl]-2-(lH-indol-3-yl)ethyl]-2-amino-2-methylpropanamide;
S) N- [ 1 (R)- [(2-Acetyl- 1,2,3 ,4-tetrahydrospiro[isoquinolin-4,4'-
piperidin]- 1 '-yl)carbonyl]-2-(indol-3-yl)ethyl]-2-amino-2-methyl-
prop~n~lnide;
6) N- [ 1 (R)-[( 1 ,2-Dihydro- 1 -methanesulfonylspiro~3H-indole-3 ,4'-
piperidin]- 1 '-yl)carbonyl]-2-(phenylmethyloxy)ethyl]-2-amino-2-
methylprop~n~mide;
20 7) N- [ 1 (R)- [( 1 ,2-Dihydro- 1 -methanesulfonylspiro[3H-indole-3 ,4'-
piperidin]- 1 '-yl)carbonyl]-2-(phenylmethyloxy)ethyl]-2-amino-2-
methylprop;~n~mide methanesulfonate;
8) N-[l(R)-[(1,2-Dihydro-1-methanesul~onylspiro[3H-indole-3,4'-
25 piperidin]- 1 I-yl)carbonyl]-2-(2',6'-difluorophenylmethyloxy)ethyl]-2-
amino-2-methylprop~n~n~ide;
9) N-Ll(R)-[(1,2-Dihydro-l-methanesulfonyl-S-fluorospiro~3H-indole-
3,4'-piperidin]- 1 '-yl)carbonyl]-2-(phenylmethyloxy)ethyl]-2-amino-2-
30 methylprop~n~mide;
10) N-[l(S)-[(1,2-Dihydro-l-meth~nesulfonylspiro[3H-indole-3,4'-
piperidin]-l'-yl) carbonyl]-2-(phenylmethylthio)ethyl]-2-amino-2-
methylprop~n~mide;
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- 20 -
11) N-[l(R)-[(1,2-Dihydro-l-methanesulfonylspiro[3H-indole-3,4'-
piperidin]- 1 '-yl)carbonyl]-3-phenylpropyl]-2-amino-2-methyl-
prop~n~mide;
12) N-[l(R)-[(1,2-Dihydro-l-methanesulfonylspiro[3H-indole-3,4'-
piperidin]- 1 '-yl)carbonyl]-3-cyclohexylpropyl]-2-amino-2-methyl-
prop~n~mide;
10 13 ) N- [ 1 (R)- [( 1 ,2-Dihydro- 1 -methanesulfonylspiro [3H-indole-3 ,4'-
piperidin]- 1 '-yl)carbonyl]-4-phenylbutyl]-2-amino-2-methyl-
propAn~mide;
14) N-[l(R)-[(1,2-Dihydro-l-methanesulfonylspiro[3H-indole-3,4'-
15 piperidin]-1'-yl)carbonyl]-2-(5-fluoro-lH-indol-3-yl)ethyl]-2-amino-2-
methylprop~n~mide;
15) N-[l(R)-[(1,2-Dihydro-l-methanesulfonyl-5-fluorospiro[3H-indole-
3,4'-piperidin]- 1 '-yl)carbonyl]-2-(5-fluoro- lH-indol-3-yl)ethyl]-2-amino-
20 2-methylpropanamide;
16) N-[l(R)-[(1,2-Dihydro-1-(2-ethoxycarbonyl)methylsulfonylspiro-
[3H-indole-3,4'-piperidin]- 1 '-yl)carbonyl]-2-( 1 H-indol-3-yl)ethyl]-2-
amino-2-methylpropanamide;
17) N-[l(R)-[(1,2-Dihydro-l,l-dioxospiro[3H-benzothiophene-3,4'-
piperidin]- 1 '-yl)carbonyl]-2-(phenylmethyloxy)ethyl]-2-amino-2-
methylprop~n~mide;
30 and pharmaceutically acceptable salts thereof.
Expecially preferred growth hormone secretagogues within
this third class which may be employed in the present invention include:
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- 21 -
N- [ 1 (R)- [( 1 ,2-dihydro- 1 -methanesulfonylspiro[3H-indole-3 ,4'-piperidin] -
1 '-yl)carbonyl] -2-(phenylmethyloxy)ethyl~ -2-amino-2 -
methylprop~n~mide;
S N-[l(R)-[(1,2-dihydro-1-methanesulfonylspiro[3H-indole-3,4'-piperidin]- 1 '-yl)carbonyl]-2-(phenylmethyloxy)ethyl~-2-amino-2-
methylprop~n~mide methanesulfonate;
and pharmaceutically acceptable salts thereof.
The most preferred compounds within this third class which
may be employed in the present invention are identified as having the
following structure:
~/\'0 C~ ICI NH2
~--N?SO2-CH3
15 and pharmaceutically acceptable salts thereof, in particular, the
methanesulfonate salt.
A representative fourth class of growth hormone
secretagogues is disclosed in U.S. Patent No. 5,492,916 as being
compounds of the structural formula I:
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- 22 -
H H O R4
Rl~N--C-A--N~
C=O R5
(CH2jn ~
~X
y
Formula I
wherein the various substituents are as defined in U.S. Patent 5,492,916.
In the above structural formulas and throughout the instant
specification, the following terms have the indicated me~nin~.c:
The alkyl groups specified above are intended to include
those alkyl groups of the designated length in either a straight or branched
10 configuration which may optionally contain double or triple bonds.
Exemplary of such alkyl groups are methyl, ethyl, propyl, ethinyl,
isopropyl, butyl, sec-butyl, tertiary butyl, pentyl, isopentyl, hexyl,
isohexyl, allyl, propenyl, butenyl, butadienyl and the like. The alkoxy
groups specified above are intended to include those alkoxy groups of the
15 designated length in either a straight or branched configuration which
may optionally contain double or triple bonds. Exemplary of such alkoxy
groups are methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy,
tertiary butoxy, pentoxy, isopentoxy, hexoxy, isohexoxy allyloxy,
propinyloxy, isobutenyloxy, 2-hexenyloxy, and the like. The term
20 "halogen" is intended to include the halogen atom fluorine, chlorine,
bromine and iodine. The term "aryl" is intended to include phenyl and
naphthyl and aromatic residues of 5- and 6- membered rings with 1 to 3
heteroatoms or fused S or 6 membered bicyclic rings with 1 to 3
heteroatoms of nitrogen, sulfur or oxygen. Examples of such heterocyclic
25 aromatic rings are pyridine, thiophene, benzothiophene, tetrazole, indole,
N-methylindole, dihydroindole, indazole, N-formylindole,
benzimidazole, thiazole, furan, pyrimidine, and thi~ 7.ole.
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WO 97/41879 PCT/US97/07516 --
Certain of the above defined terms may occur more than
once in the above formula and upon such occurrence each term shall be
defined independently of the other. Similarly, the use of a particular
variable within a noted structural formula is intended to be independent
5 of the use of such variable within a different structural formula.
Por use in medicine, the salts of the compounds of this
invention refer to non-toxic "pharmaceutically acceptable salts." Other
salts may, however, be useful in the preparation of the compounds
according to the invention or of their pharmaceutically acceptable salts.
10 Salts encompassed within the term "pharmaceutically acceptable salts"
refer to non-toxic salts of the compounds of this invention which are
generally prepared by reacting the free base with a suitable organic or
inorganic acid. Representative salts include the following: Acetate,
Benzenesulfonate, Benzoate, Bicarbonate, Bisulfate, Bitartrate, Borate,
15 Bromide, Calcium, Camsylate, Carbonate, Chloride, Clav--lAnAte, Citrate,
Dihydrochloride, Edetate, Edisylate, Estolate, Esylate, Fumarate,
Gluceptate, Gluconate, Gll-t~m~e, Glycollylars~ te, Hexylresorcinate,
HydrAbAmine, Hydrobromide, Hydrochloride, Hydroxynaphthoate,
Iodide, Isothionate, Lactate, Lactobionate, Laurate, Malate, Maleate,
20 Mandelate, Mesylate, Methylbromide, Methylnitrate, Methylsulfate,
Mucate, Napsylate, Nitrate, N-methylglucamine ammonium salt, Oleate,
Oxalate, Pamoate (Embonate), Palmitate, Pantothenate,
Phosphate/diphosphate, PolygAlActl~ronate, Salicylate, Stearate,
Subacetate, Succinate, Sulfate, Sulfonate, Tannate, Tartrate, Teoclate,
25 Tosylate, Triethiodide and Valerate. ~urthermore, where the compounds
of the invention carry an acidic moiety, suitable pharmaceutically
acceptable salts thereof may include alkali metal salts, e.g., sodium or
potassium salts; AlkAline earth metal salts, e.g., calcium or magnesium
salts; and salts formed with suitable organic ligands, e.g., quaternary
30 ammonium salts.
The compounds employed in the present invention, may
have chiral centers and occur as racemates, racemic mixtures and as
individual diastereomers, or enantiomers with all isomeric forms being
included in the present invention. Therefore, where a compound is chiral,
.. . .... ..
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- 24-
the separate enantiomers, substantially free of the other, are included
within the scope of the invention; further included are all mixtures of the
two enantiomers.
Full descriptions of the prepa~ation of the growth hormone
S secretagoue employed in the present invention may be found e.g., in:
U.S. Patent No. 3,239,345; U.S. Patent No. 4,036,979; U.S. Patent No.
4,411,890; U.S. Patent No. 5,206,235; U.S. Patent No. 5,283,241; U.S.
Patent No. 5,284,841; U.S. Patent No. 5,310,737; U.S. Patent No.
5,317,017; U.S. Patent No. 5,374,721; U.S. Patent No. 5,430,144; U.S.
10 Patent No. 5,434,261; U.S. Patent No. 5,438,136; U.S. Patent No.
5,494,919; U.S. Patent No. 5,494,920; U.S. Patent No. 5,492,916; U.S.
Patent No. 5,536,716; EPO Patent Pub.No. 0,144,230; EPO Patent Pub.
No. 0,513,974; PCT Patent Pub. No. WO 89/07110; PCT Patent Pub. No.
WO 89/07111; PCT Patent Pub.No. W O 93/04081; PCT Patent Pub. No.
15 WO 94/07486; PCT Patent Pub. No. WO 94/08583; PCT Patent Pub. No.
WO 94/11012; PCT Patent Pub. No.WO 94/13696; PCT Patent Pub.No.
WO 94/19367; PCT Patent Pub. No.WO 95/03289; PCT Patent Pub.No.
WO 95/03290; PCT Patent Pub. No.WO 95/09633; PCT Patent Pub.No.
WO 95/11029; PCT Patent Pub. No.WO 95/12598; PCT Patent Pub. No.
20 WO 95/13069; PCT Patent Pub. No. WO 95/14666; PCT Patent Pub. No.
WO 95/16675; PCT Patent Pub. No. WO 95/16692; PCT Patent Pub.No.
WO 95/17422; PCT Patent Pub. No. WO 95/17423; PCT Patent Pub.No.
WO 95/34311; PCT Patent Pub. No.WO 96/02530; PCT Patent Pub. No.
WO 96/05195; PCT Patent Pub. No.WO 96/15148; PCT Patent Pub. No.
25 WO 96/22782; PCT Patent Pub. No. WO 96/22997; PCT Patent Pub.No.
WO 96/24580; PCT Patent Pub. No. WO 96/24587; PCT Patent Pub. No.
WO 96/35713; PCT Patent Pub. No. WO 96/38471; PCT Patent Pub.No.
WO 97/00894; PCT Patent Pub. No. WO 97/06803; PCT Patent Pub. No.
WO 97/07117; J. Endocrinol Invest., lS(Suppl 4), 45 (1992)); Science.
30 ~, 1640-1643 (June 11, 1993); Ann. Rep. Med. Chem., 28, 177-186
(1993); Bioorg. Med. Chem. Ltrs., 4(22), 2709-2714 (1994); and Proc.
Natl. Acad. Sci. USA 92, 7001-7005 (July 1995), as well as herein.
Methods to obtain the growth horrnone releasing peptides
GHRP-6 and GHRP- 1 are described in U.S . Patent Nos. 4,411,890 and
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WO 97/41879 PCT/US97/07516 --
PCT Patent Publications WO 89/07110, WO 89/07111, methods to obtain
the growth hormone releasing peptide GHRP-2 are described in PCT
Patent Publication WO 93/04081, and methods to obtain hexarelin are
described in J. Endocrinol Invest., 15(Suppl 4), 45 (1992).
The identification of a compound as a growth hormone
secretagogue and thus able to directly or indirectly stimulate or increase
the endogenous release of growth hormone in an ~nim~l may be readily
determined without undue experimentation by methodology well known
in the art, such as the assay described by Smith, et al., Science, 260,
1640-1643 (1993) (see text of Figure 2 therein). In a typical experiment
pituitary glands are aseptically removed from 150-200 g Wistar male rats
and cultures of pituitary cells are prepared according to Cheng et al.
Endocrinol., 124, 2791-2798 (1989). The cells are treated with the
subject compound and assayed for growth hormone secreting activity and
intracellular cAMP levels as described by Chang et al. In particular, the
intrinsic growth horomone secretagogue activity of a compounds which
may be used in the present invention may be detennined by this assay.
The term "therapeutically effective amount" shall mean that
amount of a drug or pharmaceutical agent that will elicit the biological or
medical response of a tissue, system, ~nimzll or human that is being
sought by a researcher or clinician.
Accordingly, the present invention includes within its scope
the use of a growth hormone secretagogue, alone or in combination with
other agents, for the prevention or treatment of sleep disorders and sleep
disturbances in a warm-blooded ~nim~l. For the purposes of this
disclosure, a warm-blooded ~nim~l is a member of the ~nim~l kingdom
which includes but is not limited to m~mm~l~ and birds. The preferred
m~nnm~l for purposes of this invention is human.
Included within the scope of the present invention is the
method of using a growth hormone secretagogue for enhancing and
improving the quality of sleep. The growth hormone secretagogue is
useful in enhancing or improving sleep quality as well as preventing and
treating sleep disorders and sleep disturbances in a warm-blooded ~nim~l
In addition, the use of the growth hormone secretagogue increases sleep
.... ,, .. ,, . , . , ~ ~ .
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WO 97/41879 PCT/US97/07516
- 26 -
efficiency and augments sleep m~intenance. The growth hormone
secretagogue may further be used in a a method for preventing and
treating sleep disorders and sleep disturbances in a warm-blooded ~nim~l
The present invention further provides a pharmaceutical composition for
enhancing or improving sleep quality and increasing sleep efficiency and
sleep m~intenance.
The present method of using a growth hormone secretagogue
further provides the following: an increase in the value which is
calculated from the time that a subject sleeps divided by the time that a
subject is attempting to sleep; a decrease in sleep latency (the time it
takes to fall asleep); a decrease in difficulties in falling asleap; a decrease
in the number of awakenings during sleep; a decrease in nocturnal
arousals; a decrease in the time spent awake following the initial onset of
sleep; an increase in the total amount of sleep; an increase the amount and
~5 percentage of REM sleep; an increase in the duration and occurrence of
REM sleep; a reduction in the fragmentation of REM sleep; an increase in
the amount and percentage of slow-wave (i.e. stage 3 or 4) sleep; an
increase in the amount and percentage of stage 2 sleep; an enhancement
of EEG-delta activity during sleep; a decrease in the number of
awakenings; a decrease in nocturnal arousals, especially early morning
awakenings; an increase in daytime alertness; an increased satisfaction
with the intensity of sleep; and increased sleep maintenance. Secondary
outcomes which may be provided by the present invention include
enhanced cognitive function and increased memory retention.
The present invention is further useful for the prevention and
treatment of sleep disorders and sleep disturbances including: sleep
problems associated with insomnia, hypersomnia, sleep apnea,
narcolepsy, nocturnal myoclonus, REM sleep interruptions, jet-lag, shift
workers' sleep disturbances, dysomnias, night terror, insomnias associated
with depression or with emotionaVmood disorders, as well as sleep
walking and enuresis, as well as sleep disorders which accompany aging,
conditions associated with circadian rhythmicity, mental and physical
disorders associated with travel across time zones and with rotating shift-
work schedules, or syndromes such as fibromyalgia which are manifested
CA 022~3390 1998-10-29
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- 27 -
by non-restorative sleep and muscle pain or sleep apnea which is
associated with respiratory disturbances during sleep.
This particular application of growth hormone secretagogues
provides unexpected benefit relative to the ~rlmini.~tration of exogenous
5 growth hormone. In particular, the growth hormone secretagogue
enhances the normal pulsatile releases of endogenous growth hormone or
growth hormone-releasing hormone and thus is more likely to reproduce
the natural pattern of endogenous growth hormone release, especially
with regard to increasing the level of endogenous growth hormone prior
10 to or in during the initial onset of sleep. Growth hormone secregagogues
which are orally active also have the benefit being able to be ~f~mini.stered
orally, rather than just intravenously, intraperitoneally or subcutaneously.
Although the specific mech~ni.cm underlying the present invention is not
currently understood, it may be possible that the growth hormone
15 secretagogue not only stimulates the production of growth hormone
which provides benefical outcomes in sleep, but it also acts to increase
the levels of growth hormone-releasing hormone and/or somatostatin
which then directly enhance the quality of sleep.
In addition, the present invention includes within its scope a
20 pharmaceutical composition for enhancing and improving the quality of
sleep comprising, as an active ingredient, at least one growth hormone
secretagogues in association with a pharrnaceutical carrier or diluent.
Optionally, the active ingredient of the pharmaceutical compositions can
comprise an anabolic agent in addition to at least one growth hormone
25 secretagogue or another composition which exhibits a different activity,
e.g., an antibiotic growth promoting agent or in combination with a
corticosteroid to minimi7e the catabolic side effects or with other
ph~rm~ceutically active materials wherein the combination enhances
efficacy and minimi7es side effects. Growth promoting and anabolic
30 agents include, but are not limited to, TRH, diethylstilbesterol, estrogens,
~-agonists, theophylline, anabolic steroids, dehydroepiandrosterone,
enkephalins, E series prostaglandins, retinoic acid, compounds disclosed
in U.S. Patent No. 3,239,345, e.g., zeranol, and compounds disclosed in
U.S. Patent No. 4,036,979, e.g., sulbenox. or peptides disclosed in U.S.
.. . ..
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- 28 -
Patent No. 4,411,890. Simil~rly, the growth hormone secretagogue may
be ~lmini.~tered with somatomedin-C for the treatment of disorders
characterized by non-restorative sleep, such as f1bromyalgia (U.S. Patent
No. 5,378,686).
The present invention further includes the use of a growth
hormone secretagogue in the manufacture of a medicament for enhancing
and improving the quality of sleep and for the treatement of sleep
disorders and sleep disturbances.
In addition, the present invention contemplates the use of a
10 growth hormone secretagogue for enhancing and improving the quality of
sleep in combination with another growth hormone secretagogues such as
those referenced herein, including the growth hormone releasing peptides
GHRP-6 and GHRP-1 (described in U.S. Patent No. 4,411,890 and PCT
publications WO 89/07110, WO 89tO7111) and GHRP-2 (described in
15 WO 93/04081) and B-HT920, as well as hexarelin or growth hormone
releasing hormone (GHRH, also designated GRF) and its analogs, or
growth hormone and its analogs, or somatomedins including IGF- 1 and
IGF-2, or with a-adrenergic agonists such as clonidine or serotonin
5HTD agonists such as sumatriptan, or agents which inhibit somatostatin
20 or its release such as physostigmine and pyridostigmine. For example, a
growth hormone secretagogue may be used in combination with IGF-1
for enhancing and improving the quality of sleep.
It will be known to those skilled in the art that there are
numerous compounds now being used in an effort to enhance and
25 improve the quality of sleep. Combinations of these therapeutic agents
some of which have also been mentioned herein with a growth hormone
secretagogue will bring additional, complementary, and often synergistic
properties to enh~nce the desirable properties of these various therapeutic
agents. In these combinations, the growth hormone secretagogue and the
30 therapeutic agents may be independently present in dose ranges from one
one-hundredth to one times the dose levels which are effective when
these compounds and secretagogues are used singly.
The growth hormone secretagogue may be ~trnini.~tered in
combination with sedatives, hypnotics, anxiolytics, antipsychotics,
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- 29 -
antianxiety agents, minor tran~uilizers, melatonin agonists and
antagonists, melatonergic agents, benzodiazepines, barbituates, 5HT-2
antagonists, and the like, or the growth hormone secretagogue may be
A~lmini~tered in conjunction with the use of physical methods such as
5 with light therapy or electrical stimulation. For example, to enhance and
improve the quality of sleep a growth hormone secretagogue may be
given in combination with such compounds as: ~-linA701am, allobarbital,
alonimid, alprazolam, amitriptyline, amobarbital, amoxapine,
bentazepam, benzoctamine, brotizolam, bupropion, busprione,
10 butabarbital, butalbital, capuride, carbocloral, chloral betaine, chloral
hydrate, chlordiazepoxide, clomipramine, cloperidone, clorazepate,
clorethate, clozapine, cyprazepam, desipramine, dexclamol, diazepam,
dichloralphenazone, divalproex, diphenhydramine, doxepin, estazolam,
ethchlorvynol, etomidate, fenobam, flunitrazepam, flurazepam,
15 fluvoxamine, fluoxetine, fosazepam, glutethimide, halazepam,
hydroxyzine, imipramine, lithium, lorazepam, lormetazepam,
maprotiline, mecloqualone, melatonin, mephobarbital, meprobamate,
methaqualone, midaflur, midazolam, nefazodone, nisobamate,
nitrazepam, nortriptyline, oxazepam, paraldehyde, paroxetine1
20 pentobarbital, perlapine, perphenazine, phenelzine, phenobarbital,
prazepam, prometh~7ine, propofol, protriptyline, quazepam, reclazepam,
roletamide, secobarbital, sertraline, suproclone, temazepam, thioridazine,
tracazolate, tranylcypromaine, trazodone, triazolam, trepipam,
tricetamide, triclofos, trifluoperazine, trimetozine, trimipramine,
25 uldazepam, venlafaxine, zaleplon, zolazepam, zolpidem, and salts
thereof, and combinations thereof, and the like, as well as admixtures and
combinations thereof.
Typically, the individual daily dosages for these
combinations may range from about one-fifth of the minim~lly
30 recommended clinical dosages to the maximum recommended levels for
the entities when they are given singly.
To illustrate these combinations, a growth hormone
secretagogue effective clinically effective clinically at a given daily dose
range may be effectively combined, at levels which are equal or less than
. ... ~ ~............ .. . .
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- 30-
the daily dose range, with the following compounds at the indicated per
day dose range: adinazolam, allobarbital, alonimid, alprazolam,
amitriptyline, amobarbital, amoxapine, bentazepam, benzoctamine,
brotizolam, bupropion, busprione, butabarbital, butalbital, capuride,
S carbocloral, chloral betaine, chloral hydrate, chlordiazepoxide,
clomipramine, cloperidone, clorazepate, clorethate, clozapine,
cyprazepam, desipramine, dexclamol, diazepam, dichloralphenazone,
divalproex, diphenhydramine, doxepin, estazolam, ethchlorvynol,
etomidate, fenobam, flunitrazepam, flurazepam, fluvoxamine, fluoxetine,
fosazepam, glutethimide, halazepam, hydroxyzine, imipramine, lithium,
lorazepam, lormetazepam, maprotiline, mecloqualone, melatonin,
mephobarbital, meprobamate, methaqualone, midaflur, midazolam,
nefazodone, nisobamate, nitrazepam, nortriptyline, oxazepam,
paraldehyde, paroxetine, pentobarbital, perlapine, perphenazine,
phenelzine, phenobarbital, prazepam, promethazine, propofol,
protriptyline, quazepam, reclazepam, roletamide, secobarbital, sertraline,
suproclone, temazepam, thioridazine, tracazolate, tranylcypromaine,
trazodone, triazolam, trepipam, tricetamide, triclofos, trifluoperazine,
trimetozine, trimipramine, uldazepam, venlafaxine, zaleplon, zolazepam,
zolpidem, and salts thereof, and combinations thereof, and the like, as
well as admixtures and combinations thereof. It will be readily apparent
to one skilled in the art that the growth hormone secretagogue may be
employed with other agents to control sleep disorders and sleep
disturbances in depressed patients and/or provide benefit in the
prevention or treatment of sleep disorders and sleep disturbances.
Naturally, these dose ranges may be adjusted on a unit basis
as necessary to permit divided daily dosage and, as noted above, the dose
will vary depending on the nature and severity of the disease, weight of
patient, special diets and other factors.
Anabolic effects especially in the treatment of geriatric male
patients are obtained with compounds of this invention in combination
with anabolic steroids such as dehydroepiandrosterone, oxymetholone,
methyltesterone, fluoxymesterone, restosterone and stanozolol.
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These combinations may be formulated into pharmaceutical
compositions as known in the art and as discussed below. A growth
hormone secretagogue may be ~1mini~tered alone or in combination by
oral, parenteral (e.g., intramuscular, intraperitoneal, intravenous or
5 subcutaneous injection, or implant), nasal, v~gin~l, rectal, sublingual, or
topical routes of ~(lmini.~tration and can be formulated in dosage forms
apyloyliate for each route of ~-lmini~tration.
Solid dosage forms for oral ~clmini~tration include capsules,
tablets, pills, powders and granules. In such solid dosage forrns, the
10 active compound is admixed with at least one inert pharmaceutically
acceptable carrier such as sucrose, lactose, or starch. Such dosage forms
can also comprise, as is normal practice, additional substances other than
inert diluents, e.g., lubricating agents such as magnesium stearate.
Illustrative of the adjuvants which may be incorporated in tablets,
15 capsules and the like are the following: a binder such as gum tr~g;~c~nth,
acacia, corn starch or gelatin; an excipient such as microcrystalline
cellulose; a disintegrating agent such as corn starch, pregel~tini7ed starch,
alginic acid and the like; a lubricant such as magnesium stearate; a
sweetening agent such as sucrose, lactose or saccharin; a flavoring agent
20 such as peppermint, oil of wintergreen or cherry. In the case of capsules,
tablets and pills, the dosage forms may also comprise buffering agents.
When the dosage unitform is a capsule, it may contain, in addition to
materials of the above type, a liquid carrier such as fatty oil. Various
other materials may be present as coatings or to otherwise modify the
25 physical form of the dosage unit. Tablets and pills can additionally be
prepared with enteric coatings and tablets may be coated with shellac,
sugar or both.
Liquid dosage forms for oral ~-lmini.~tration include
pharmaceutically acceptable emulsions, solutions, suspensions, syrups,
30 the elixirs containing inert diluents commonly used in the art, such as
water. Besides such inert diluents, compositions can also include
adjuvants, such as wetting agents, emulsifying and suspending agents,
and sweetening, flavoring, and perfuming agents. A syrup or elixir may
contain the active compound, sucrose as a sweetening agent, methyl and
, . . , ., . ~ , ~ ....... , ~ . .......
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propyl parabens as preservatives, a dye and a flavoring such as cherry or
orange flavor.
Preparations according to this invention for parenteral
~dmini.stration include sterile aqueous or non-aqueous solutions,
5 suspensions, or emulsions. Sterile compositions for injection may be
formulated according to conventional pharmaceutical practice by
dissolving or suspending the active substance in a vehicle such as water
for injection, a naturally occurring vegetable oil like sesame oil, coconut
oil, peanut oil, cottonseed oil, etc., or a synthetic fatty vehicle like ethyl
10 oleate or the like. Buffers, preservatives, antioxidants and the like may
be incorporated as required. Examples of non-aqueous solvents or
vehicles are propylene glycol, polyethylene glycol, vegetable oils, such as
olive oil and corn oil, gelatin, and injectable organic esters such as ethyl
oleate. Such dosage forms may also contain adjuvants such as
15 preserving, wetting, emulsifying, and dispersing agents. They may be
sterilized by, for example, filtration through a bacteria-retaining filter, by
incorporating sterilizing agents into the compositions, by irr~ ting the
compositions, or by he~tin.~ the compositions. They can also be
manufactured in the form of sterile solid compositions which can be
20 dissolved in sterile water, or some other sterile in3ectable medium
immediately before use. Compositions for rectal or vaginal
~lmini~tration are preferably suppositories which may contain, in
addition to the active substance, excipients such as cocoa butter or a
suppository wax. Compositions for nasal or sublingual ~limini.~tration are
25 also ~lc~al~;d with standard excipients well known in the art.
The dosage of active ingredient in the compositions of this
invention may be varied, however, it is necessary that the amount of the
active ingredient be such that a suitable dosage form is obtained. The
active ingredient may be ~tlmini~tered to patients (~nim~ls and human) in
30 need of such treatment in dosages that will provide optimal
pharmaceutical efficacy. The selected dosage depends upon the desired
therapeutic effect, on the route of ~dministration, and on the duration of
the treatment. The dose will vary from patient to patient depending
upon the nature and severity of disease, the patient's weight, special diets
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then being followed by a patient, concurrent medication, and other factors
which those skilled in the art will recognize. Generally, dosage levels of
between 0~0001 to 10 mg/kg. of body weight daily are ~rlmini~tered to
patients and ~nim~l~, e.g., m~mm~ , to obtain effective release of growth
5 hormone. The dosage range will generally be about 0~5 mg to 1.0 g. per
patient per day which may be ~Amini~tered in single or multiple doses.
Perferably, the dosage range will be about 0.5 mg to 500 mg per patient
per day; more preferably about 0.5 mg to 200 mg per patient per day; and
even more preferably about 5 mg to 50 mg per patient per day.
Pharmaceutical compositions of the present invention may
be provided in a solid dosage formulation preferably comprising about
0.5 mg to 500 mg active ingredient, more preferably comprising about 1
mg to 250 mg active ingredient. The pharmaceutical composition is
preferably provided in a solid dosage formulation comprising about 1 mg,
15 5 mg, 10 mg, 25 mg, 50 mg, 100 mg, 200 mg or 250 mg active
ingredient.
The following examples are provided for the purpose of
further illustration only and are not intended to be limitations on the
disclosed invention.
EXAMPLE 1
3-Amino-3-methyl-N-[2,3,4,5-tetrahydro-2-oxo- 1-[~2'-(1 H-tetrazol-5-
yl)r 1~1 '-biphenyll-4-yllmethyll- lH- 1 -benzazepin-3(R)-yll-b-lt~n~rnide,
Step A: 3-Amino-2.3.4.5-tetrahydro- 1 H- 1 -benzazepin-2-one
A solution of 9.22 g (45.6 mmol) of 3-azido-2,3,4,5-
tetrahydro-lH-1-benzazepin-2-one (prepared by the method of Watthey,
et al., J. Med. Chem., 28, 1511-1516 (1985)) in 30 mL methanol was
30 hydrogenated at 40 psi in the presence of 1.0 g of 5% Pt/C for 4.5 hours.
Celite was added and the mixture filtered through a pad of Celite. The
filtrate was concentrated and allowed to stand for 16 hours at room
temperature which resulted in formation of crystals. The material was
isolated by filtration and dried under vacuum to afford 4.18 g (23.7
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- 34-
mmol, 52%) of the product. The mother liquors were diluted to 100 mL
with methanol, treated with 2 g of charcoal, filtered through Celite and
the filtrate concentrated under vacuum to approximatley 15 mL. A
second crop formed yielding 2.02 g of product (11.5 mrnol, 25%).
5 Another recycling of the mother liquors afforded a third crop of 0.88 g
(5.0, 11 %). A total of 7.08 g (40.2 mmol, 88%) of the product was thus
obtained. 1H NMR (200 MHz, CDCl3): 1.6 (br s, 2H), 1.80 (m, lH),
2.55 (m, 2H), 2.88 (m, lH), 3.~2 (dd; 7Hz, 1 lHz; lH), 6.98 (d, 8Hz, lH),
7.2 (m, 3H), 8.3 (br s, lH). FAB-MS: calculated for C1oH12N2O 176;
10 found 177 (M+H, 100%).
Step B: 3(R)-Amino-2~3~4~5-tetrahydro-lH-1-benzazepin-2-one
2.37 g (13.5 mmol) of 3-amino-2,3,4,5-tetrahydro-lH-1-
benzazepin-2-one (Step A) and 2.02 g (13.5 mmol) of L-tartaric acid
15 were suspended in 40 mL of ethanol. The mixture was gently heated and
complete dissolution achieved by dropwise addition of 5 mL of distilled
water. The solution was cooled to room temperature and aged overnight.
The solid that formed was removed by filtration, washed with
ethanol/diethyl ether (1:1) and dried under vacuum to afford 1.75 g of
crude L-tartrate salt. The mother liquors were evaporated to dryness
under vacuum, redissolved in 40 mL of water and the pH adjusted to 10-
11 by the addition of solid potassium carbonate. The mixture was
extracted with chloroform (6x20 mL) and the combined extracts washed
with water (1x) and brine (1x), dried over potassium carbonate, filtered
and solvents removed under vacuum to afford 1.29 g (7.33 mmol) of
partially enriched 3(R) amine. The original 1.75 g batch of L-tartrate salt
was recryst~lli7e-1 twice from aqueous ethanol to afford 1.03 g (3.17
mmol, 24~o) of purified L-tartrate salt with [a]D=-212~ (c=1, H20). The
purified L-tartrate salt was dissolved in 20 mL of water and the pH
30 adjusted to 10-11 by the addition of solid potassium carbonate. The
mixture was extracted with chloroform (SxlO mL); combined extracts
were washed with water and brine then dried over potassium carbonate,
ltered and solvents removed under vacuum to afford 522 mg (2.96
mmol, 22% overall) of the 3(S) amine, [a]D=-446o (c=l,CH30H). The
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W O 97/41879 PCT~US97/07516 --
rem~ining 1.29 g (7.33 mmol) of partially enriched 3(R) arnine was
treated with 1.10 g (7.33 mmol) of D-tartaric acid as described above and
the resulting salt recrystallized twice from aqueous ethanol to afford 1.20
g of purified D-tartrate salt, [a]D=-214~ (C=l~H2o). The purified D-
5 tartrate salt was dissolved in 20 mL of water and the free base isolated asdescribed above to give 629 mg (3.57 mmol, 26% overall) of the 3(R)
amine, [a~D=+455~ (c=l,CH30H).
Step C: 2~2-Dimethylbutanedioic acid. 4-methyl ester
2,2-dimethylsuccinic acid (20 g, 137 mmol) dissolved in 200
mL absolute methanol at 0~C was treated dropwise with 2 mL
concentrated sulfuric acid. After the addition was complete, the mixture
was allowed to warm to room temperature and stirred for 16 hours. The
mixture was concentrated in vacuo to 50 mL and slowly treated with 200
mL of saturated aqueous sodium bicarbonate. The mixture was washed
with hexane (3x) and the aqueous layer removed and cooled in an ice
bath. The mixture was acidified to pH 2 by slow addition of 6N HCl then
extracted with ether (8x). The combined extracts were washed with
brine, dried over magnesium sulfate, filtered and solvents removed in
vacuo. The residue was dried at room temperature under vacuum to
afford 14.7 g (91.8 mmol, 67%) of a viscous oil that slowly solidified
upon standing. lH NMR analysis indicates the product is a mixture of
the title compound and 15% of the isomeric 2,2-dimethylbutanedioic
acid, 1-methyl ester. NMR (200 MHz, CDC13) of title compound: 1.29
(s, 6H), 2.60 (s, 2H), 3.66 (s, 3H). NMR (200 MHz, CDC13) of isomer:
1.28 (s, 6H), 2.63 (s, 2H), 3.68 (s, 3H).
Step D: 3-[Benzyloxycarbonylamino]-3-methylbutanoic acid,
methyl ester
To 14.7 g (91.8 mmol) of 2,2-dimethylbutanedioic acid-4-
methyl ester (Step C), containing 15% of the isomeric 1-methyl ester
compound, in 150 mL benzene was added 13 mL of triethylamine (9.4 g,
93 mmol, 1.01 eq) followed by 21.8 mL diphenylphosphoryl azide (27.8
g, 101 mmol, 1.1 eq). The mixture was heated under nitrogen at reflux
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- 36-
for 45 minutes then 19 mL (19.9 g, 184 mmol, 2 eq) of benzyl alcohol
was added and refluxing continued for 16 hours. The mixture was
cooled, filtered and the filtrate concentrated to a minimum volume under
vacuum. The residue was redissolved in 250 mL ethyl acetate, washed
5 with water (lx), saturated aqueous sodium bicarbonate (2x) and brine
(lx). The organic layer was removed, dried over magnesium sulfate,
filtered and the filtrate concentrated to a minimum volume in vacuo. The
crude product was purified by medium pressure liquid chromatography
on silica, eluting with hexane/ethyl acetate (4:1), to afford 18.27 g (68.9
10 mmol, 75%) of the title compound as a pale yellow liquid in addition to a
small amount of pure 3-[benzyloxycarbonylamino]-2,2-imethylpropanoic
acid, methyl ester. lH NMR (200MHz, CDC13) of title compound: 1.40
(s, 6H), 2.69 (s, 2H), 3.63 (s, 3H), 5.05 (s, 2H), 5.22 (br s, lH), 7.32 (s,
5H). lH NMR (200 MHz, CDC13) of 3-[benzyloxycarbonylamino]-2,2-
15 dimethylpropanoic acid, methyl ester (200 MHz, CDC13): 1.19 (s, 6H),3.30 (d, 7Hz, 2H; resonance collapses to singlet in CD30D), 3.67 (s, 3H),
5.09 (s, 2H), 5.22 (br s,lH; resonance not observed in CD30D), 7.3 (br s,
SH).
20 StepE: 3-Benzyloxycarbonylamino-3-methylbutanoic acid
A solution of 18.27 g (68.9 mmol) of methyl 3-
benzyloxycarbonylamino-3-methylbutanoate (Step D) in 20 mL of
methanol at room temperature was treated dropwise with 51 mL of 2N
NaOH (102 mmol, 1.5 eq). The mixture was stirred at room temperature
25 for 16 hours then transferred to a separatory funnel and washed with
hexane (3x). The aqueous layer was removed, cooled to 0~C and slowly
acidi~led to pH 2 (paper) by dropwise addition of 6N HCl. This mixture
was extracted with ether (6x); combined extracts were washed with lN
HCl and brine, then dried over magnesium sulfate, filtered and solvent
30 removed under vacuum to afford 17.26 g (68.7 mmol, 99%) of the
product. lH NMR (200 MHz, CDCl3): 1.42 (s, 6H), 2.77 (s, 2H), 5.06
(s, 2H), 5.2 (br s, lH), 7.3 (s, SH).
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- 37-
Step F: 3-Benzyloxycarbonylamino-3-methyl-N-[2,3,4,5-tetrahydro-
2-oxo-lH-l-benzazepin-3(R)-yll-but~n~mide
To a solution of 252 mg (1.43 mmol) of 3(R)-amino-2,3,4,5-
tetrahydro-lH-[l]benzazepin-2-one (Step B) in 4 mL of methylene
5 chloride at room temperature was added 400 mg (1.60 rnrnol, 1.1 eq) of
3-benzyloxycarbonylamino-3-methylbutanoic acid (Step E) followed by
760 mg (1.7 mmol, 1.2 eq) benzotriazol-l-yloxytris(dimethylamino)-
phosphonium hexafluoro-phosphate and 0.50 mL of diisopropyl-
ethylamine (380 mg, 2.9 mmol, 2 eq). After 3 hours at room temperature,
10 the mixture was diluted into 30 mL of ethyl acetate and washed with 5%
aqueous citric acid, saturated aqueous sodium bicarbonate (2x) and brine.
The organic layer was removed, dried over magnesium sulfate, filtered
and solvents removed under vacuum. The residue was purified by
medium pressure liquid chromatography on silica, eluting with ethyl
15 acetate to afford 586 mg (1.43 mmol, 100%) of the product. 1H NMR
(200 MHz, CDCl3): 1.38 (s, 3H), 1.39 (s, 3H), 1.82 (m, lH), 2.52 (s,
2H), 2.5-3.0 (m, 3H), 4.51 (m, lH), 5.07 (br s, 2H), 5.57 (br s, lH), 6.68
(d, 7Hz, lH), 6.97 (d, 8Hz, lH), 7.1-7.4 (m, 8H), 7.61 (br s, lH). FAB-
MS: calculated for C23H27N304 409; found 410 (M+H, 100%);
20 [a]D=+137~ (c=1, CHCl3).
Step G: 5-Phenyltetrazole
Zinc chloride (3.3 g, 24.3 mmol, 0.5 eq) was added to 15 mL
of N,N-dimethylformamide in small portions while m~i"t~ining the
25 temperature below 60~C. The suspension of zinc chloride was cooled to
room temperature and treated with 5.0 g of benzonitrile (48.5 mmol, 1.0
eq) followed by 3.2 g of sodium azide (48.5 mmol, 1.0 eq). The
heterogeneous mixture was heated at 115~C with agitation for 18 hours.
The mixture was cooled to room temperature, water (30 mL) was added
30 and the mixture acidified by the addition of 5.1 mL of concentrated
hydrochloric acid. The mixture was cooled to 0~C and aged for one hour,
then filtered and the filter cake washed with 15 mL of cold O.lN HCl then
dried at 60~C under vacullm to afford 6.38 g (43.7 mmol, 90%) of the
product.
.. . ..
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Step H: 5-Phenyl-2-trityltetrazole
To a suspension of 5.0 g (34.2 mmol) of 5-phenyltetrazole in
55 mL of acetone was added 5.0 mL of triethylamine (3.6 g, 35.6 mmol,
1.04 eq). After 15 minutes, a solution of 10.0 g of triphenyl-methyl
5 chloride (35.9 mmol, 1.05 eq) in 20 mL of tetrahydrofuran was added and
the mixture stirred at room temperature for one hour. Water (75 mL) was
slowly added and the mixture stirred for one hour at room temperature.
The product was collected by filtration, washed with 75 mL of water and
dried at 60~C under vacuum to give 13.3 g (34.2 mmol, 100%) of the
10 product.
Step I: N-Triphenylmethyl-5-r2-(4'-methylbiphen-4-yl)~ tetrazole
A solution of zinc chloride (6.3 g, 46.2 mmol, 0.6 eq) in 35
mL of tetrahydrofuran was dried over molecular sieves. 5-Phenyl-2-
15 trityltetrazole (30.0 g, 77.3 mmol, 1.0 eq) was dissolved in 300 mL of drytetrahydrofuran and the solution gently stirred while being degassed three
times by alternating vacuum and nitrogen purges. The stirred solution
was cooled to -15~C and treated slowly with 50.5 mL of 1.6 M n-
butyllithium in hexane (80.0 mmol, 1.05 eq) so as to m~int~in the
20 temperature below -5~C. The solution was maintained at -5 to -15~C for
1.5 hours then treated with the dried zinc chloride solution and allowed to
warm to room temperature. In a separate flask, 4-iodotoluene (20.17 g,
92.5 mmol, 1.2 eq) and bis-(triphenylphosphine)nickel (II) dichloride (1.5
g, 2.3 mmol, 0.03 eq) were dissolved in 60 mL of tetrahydrofuran, then
25 degassed and left under an atmosphere of nitrogen. The mixture was
cooled to 5~C and treated with 1.5 mL of 3.0 M solution of
methylmagnesium chloride in tetrahydrofuran (4.5 mmol, 0.06 eq) so as
to keep the temperature below 10~C. The solution was warmed to room
temperature and added, under nitrogen purge, to the arylzinc solution.
30 The reaction mixture was stirred vigorously for 8 hours at room
temperature then quenched by the slow addition of a solution of 10 mL of
glacial acetic acid (1.6 mmol, 0.02 eq) in 60 mL of tetrahydrofuran at a
rate so that the temperature was m~int~ined below 40~C. The mixture
was stirred for 30 minutes and 150 mL of 80% saturated aqueous sodium
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- 39-
chloride was ~lAe-l; the reaction mixture was extracted for 30 minutes
and the layers allowed to separate. The organic layer was removed and
washed with 150 mL of 80% saturated aqueous sodium chloride buffered
to pH>lO by the addition of ammonium hydroxide. The organic phase
S was removed and concentrated under vacuum to approximately 50 mL
then 250 mL of acetonitrile was added. The mixture was again
concentrated under vacuum to 50 mL and acetonitrile added to make the
final volume 150 mL. The resulting slurry was cooled at 5~C for 1 hour
then filtered and washed with S0 mL of cold acetonitrile followed by 150
10 mL of distilled water. The filter cake was air dried to a free flowing solid
then further dried under vacuum at 50~C for 12 hours to afford 30.0 g
(62.8 mmol, 81%) of the product. lH NMR (200 MHz, CDCl3): 2.28 (s,
3H), 6.9-7.05 (m, lOH), 7.2-7.5 (m, 12H), 7.9 (m, lH).
15 Step J: N-Triphenylmethyl-5-[2-(4'-bromomethylbiphen-4-yl)]
tetrazole
A solution of 3.15 g (6.6 mmol) of N-triphenylmethyl-5-[2-
(4'-methylbiphen-4-yl)] tetrazole (Step I) in 25 mL of methylene chloride
was treated with 1.29 g (7.25 mmol, 1.1 eq) of N-bromo-succinimide, 80
20 mg (0.5 mmol, 0.07 eq) of AIBN, 200 mg of sodium acetate and 200 mg
of acetic acid. The mixture was heated at reflux for 2 to 16 hours then
cooled and washed with saturated aqueous sodium bicarbonate. The
organic layer was removed, dried over sodium sulfate, filtered and
concentrated to a minimum volume by atmospheric distillation. Methyl t-
25 butyl ether was added and distillation continued until almost all themethylene chloride was removed the the total volume reduce to
approximately 12 mL and 12 mL of hexanes was then added. The
mixture was kept at room temperature for 2 hours and the product
isolated by filtration, washed with hexanes then dried under vacuum at
30 50~C to give 2.81g (5.04 mmol, 76%) of the product. 1H NMR (200
MHz, CDC13): 4.38 (s, 2H), 6.9-8.0 (m, 23H). NMR indicates presence
of approximately 1 % of the starting material and 7% of the dibromo
derivative.
, . .. . . .
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- 40 -
Step K: 3-Benzyloxycarbonylamino-3-methyl-N-[2,3,4,5-tetrahydro-
2-oxo- 1 -[[2'-(N-triphenylmethyl)-tetrazol-5-yl] [1,1 '-
biphenyl]-4-yl]methyl- 1 H- 1 -benzazepin-3(R)-yl]-
bllt~n~mide
S To a solution of 437 mg (1.07 mmol) of the intermediate
obtained in Step F in 2 mL of dry dimethylformamide at room
temperature under nitrogen was added 55 mg of 60% sodium hydride oil
dispersion (33 mg NaH, 1.38 mmol, 1.3 eq). After lS minutes, a solution
of 715 mg (1.28 mmol, 1.2 eq) N-triphenyl-methyl-5-~2-(4'-
bromomethylbiphen-4-yl)] tetrazole (Step J) in l.S mL of dry dimethyl-
formamide was added and the mixture stirred for 90 minutes. The
reaction mixture was added to 100 mL of ethyl acetate and washed with
water (2x) and brine. The organic layer was removed, dried over
magnesium sulfate, filtered and solvents removed under vacuum.
lS Purification by medium pressure liquid chromatography on silica, eluting
with ethyl acetate/hexane (1:1), afforded 902 mg (1.02 mmol, 95%) of the
product. lH NMR (200 MHz, CDCl3): 1.38 (s, 3H), 1.39 (s, 3H), 1.68
(m, lH), 2.2-2.5 (m, SH), 4.44 (m, lH), 4.67 (d, 14Hz, lH), 5.06 (s, 2H),
5.12 (d, 14Hz, lH), 5.63 (br 1, lH), 6.65 (d, 8Hz, lH), 6.9-7.5 (m, 31H),
7.85 (m, lH).
Step L: 3-Amino-3-methyl-N-[2,3,4,5-tetrahydro-2-oxo-1-[[2'-(lH-
tetrazol-S-yl] [1,1 '-biphenyl] -4-yl]methyl- 1 H- 1 -benzazepin-
3(R)-yll-but~n~mide~ trifluoroacetate
A solution of 902 mg (1.02 mmol) of the intermediate
obtained in Step H in S mL methanol was hydrogenated at room
temperature and one atmosphere over 160 mg of 20% Pd(OH)2/C for 14
hours. The mixture was filtered through Celite and concentrated under
vacuum. The residue was purified by reverse phase HPLC on C-18,
eluting with methanol/0.1 % aqueous trifluoroacetic acid (linear gradient:
60% methanol increased to 80% methanol over 10 minutes) to afford 568
mg (0.91 mmol, 89%) of the tit}e compound. 1H NMR (200 MHz,
CD30D): 1.33 (s, 3H), 1.37 (s, 3H), 2.0-2.6 (m, 6H), 4.35 (dd; 7, 11 Hz;
lH), 4.86 (d, lS Hz, lH), 5.20 (d, lS Hz, lH), 7.00 (d, 8 Hz, 2H), 7.15-
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7 35 (m, 6H), 7.45-7.70 (m, 4H). FAB-MS: calculated for C29H31N7O2
509; found 510 (M+H, 100%).
EXAMPLE 2
s
3-[(2(R)-Hydroxypropyl)amino]-3-methyl-N-[2,3,4,5-tetrahydro-2-oxo-
1-[~2'-(lH-tetrazol-5-yl)[l,l-biphenyl~-4-yl~methyl]-lH-l-benzazepin-
3(R)-yll-bllt~qn~mide
Step A: 3-[(2-(R)-Benzyloxypropyl)amino]-3-methyl-N-[2,3,4,5-
tetrahydro-2-oxo- 1 - [ [2 '-(1 H-tetrazol-5 -yl) [1,1 '-biphenyl] -4-
yl]methyl]- 1 H- 1 -benzazepin-3(R)-yl]but~n~mide,
trifluoroacetate
The title compound was prepared from 3-amino-3-methyl-N-
15 [2,3,4,5-tetrahydro-2-oxo- 1 -C[2'-( lH-tetrazol-5-yl)[ l,1 '-biphenyl~-4-
yl]methyl]- 1 H- 1 -benzazepin-3(R)-yl]but~n~mide, trifluoroacetate
(Example 1) and (O-2-benzyloxlpropanal (prepared from ethyl-D-lactate
according to the procedure of Hanessian and Kloss, Tetrahedron Lett., 26,
1261-1264 (1985) by the procedure described in U.S. Patent No.
20 5,206,235, Example 86, Step A. lH NMR (200MHz, CD30D): 1.25 (d,
6Hz, 3H), 1.35 (s, 6H), 2.11 (m, lH), 2.32 (m, lH), 2.5-2.7 (m, 4H), 2.95
(m, lH), 3.17 (m, lH), 3.80 (m, lH), 4.40 (m, lH), 4.44 (d, llHz, lH),
4.64 (d, 1 lHz, lH), 4.90 (d, l5Hz, lH), 5.02 (d, l5Hz, lH), 6.99 (d, 8Hz,
2H), 7.1-7.7 (m, 15H). FAB-MS: calculated for C39H43N703 657;
25 found 658 (M+H, 100%).
Step B: 3-[(2(R)-Hydroxypropyl)amino]-3-methyl-N-[2,3,4,5-
tetrahydro-2-oxo-1-[[2'-(lH-tetrazol-5-yl)[l,l'-biphenyl]-4-
yl]methyl]-lH-l -benzazepin-3(R)-yl]but~n~ide,
trifluoroacetate
The title compound was prepared from the intermediate
obtained in Step A by the procedure described in U.S. Patent No.
5,206,235, Example 86, Step B. lH NMR (400MHz, CD30D): 1.22 (d,
6Hz, 3H), 1.37 (s, 3H), 1.39 (s, 3H), 2.10 (m, lH), 2.31 (m, lH), 2.45-
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2.70 (m, 4H), 2.81 (dd; 10, 12Hz; lH), 3.08 (dd; 4, 12Hz; lH), 3.92 (m,
lH), 4.36 (dd; 7, llHz; lH), 4.93 (d, l5Hz, lH), S.17 (d, 15Hz, lH), 7.04
(d, 8Hz, 2H), 7.19 (d, 8Hz, 2H), 7.20-7.35 (m, 4H), 7.54 (m, 2H), 7.65
(m, 2H). FAB-MS: calculated for C32H37N7o3 567; found 568 (M+H,
5 45%)-
EXAMPLE 3 (METHOD 1)
N- [ l (R)- [(1,2-Dihydro- 1 -methanesulfonylspiro[3H-indole-3,4'-piperdin l -
10 1 '-yl)carbonyl]-2-(phenylmethyloxy)ethyl]-2-amino-2-
methylprop~n~mide
Step A: 1,2-Dihydro-l-methanesulfonylspiro[3H-indole-3,4'-
piperdinelhydrochloride
To a solution of 1.20 g (5.8mmol) of 1'-methyl-1,2-dihydro-
spiro[3H-indole-3,4'-piperdine] (prepared as described by H. Ong, et al.,
J. Med. Chem., 23, 981-986 (1983)) in 20 mL of dry dichloromethane at
0~C was added triethylamine (0.90 mL; 6.4 mmol) and methanesulfonyl
chloride (0.49 mL; 6.35 mmol) and stirred for 30 min. The reaction
20 mixture was poured into 15 mL of saturated aqueous sodium bicar~onate
solution and extracted with dichloromethane (2X10 mL). The combined
organics were washed with brine (20 mL), dried over anhydrous
potassium carbonate, filtered and the solvent removed under reduced
pressure to yield 1.44 g of the methanesulfonamide derivative as pale
25 yellow oil which was used without purification.
To a solution of above crude product in 20 mL of dry 1,2-
dichloroethane at 0~C was added 1.0 mL (9.30 mmol) of l-chloroethyl
chloroformate, and then stirred at RT for 30 min and finally at reflux for
lh. The reaction mixture was concentrated to approximately one third of
30 the volume and then diluted with 20 mL of dry methanol and refluxed for
1.5h. The reaction was cooled to RT and concentrated to approximately
one half of the volume. The precipitate was filtered and washed with a
small volume of cold methanol. This yielded 1.0 g of the piperidine HCl
salt as a white solid. The filtrate was concentrated and a small volume of
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methanol was added followed by ether. The precipitated material was
once again filtered, washed with cold methanol, and dried. This gave an
additional 0.49 g of the desired product. Total yield 1.49 g (70%).
lH NMR (CDC13, 200MHz) ~ 7.43-7.20 (m, 3H), 7.10 (dd, lH), 3.98 (bs,
5 2H), 3.55-3.40 (bd, 2H), 3.35-3.10 (m, 2H), 2.99 (s, 3H), 2.15 (t, 2H),
2.00 (t, 2H).
Step B: N-[l(R)-[(1,2-Dihydro-l-methanesulfonylspiro[3H-indole-
3,4'-piperdin]- 1 '-yl)carbonyl]-2-(phenylmethyloxy)ethyl]-2-
[(1,1 -dimethylethoxy)carbonyllamino-2-methyl-
prop~n~mlde
To 0.35g (1.15 mmol) of (2R)-2-[(1,1-dimethylethoxy)-
carbonyl]amino-3-L2-(phenylmethyloxy)ethyl]-1-propanoic acid in 13 mL
of dichloromethane was added 1,2-dihydro-1-methanesulfonylspiro-[3H-
15 indole-3,4'-piperdine] hydrochloride (0.325 g; 1.07 mmol), 0.18 mL (1.63
mmol) of N-methylmorpholine, 0.159 g (1.18 mmol) of 1-
hydroxyben7tri~7ole(HOBT) and stirred for 15 min. EDC (0.31 g; 1.62
mol) was added and stirring was continued for lh. An additional 60 ~lL
of N-methylmorpholine was added and stirred for 45 min. The reaction
20 mixture was poured into 5 mL of water and the organic layer was
separated. The organic layer was washed with 5 mL of 0.5N aqueous
hydrochloric acid and 5 mL of saturated aqueous sodium bicarbonate
solution. The combined organics were dried over anhydrous magnesium
sulfate, and concentrated to yield 0.627 g of the product as a yellow foam
25 which was used without purification.
To a 0.627 g (1.07 mmol) of the above product in 5 mL of
dichloromethane was added 1.0 mL of trifluoroacetic acid and stirred at
RT for 75 rnin. An additional 1.00 mL of trifluoroacetic acid was added
and stirred for 10 rnin. The reaction mixture was concentrated, diluted
30 with 5.0 mL of dichloromethane and carefully basified by pouring into 10
mL of 10% aqueous sodium carbonate solution. The organic layer was
separated and the aqueous layer was further extracted with 2X15 mL of
dichloromethane. The combined organics were washed with 5 mL of
water, dried over potassium carbonate, filtered and concentrated to give
... . .
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the 0.486 g of the amine as a light yellow foam which was used without
purification.
To 0.486 g (1.01 mmol) of the amine and 10 mL of
dichloromethane was added 0.26g (1.28 mmol) of 2-[(1,1-dimethyl-
5 ethoxy)carbonyl]amino-2-methyl-propanoic acid, 0.173 g (1.28 mmol) of
1-hydroxybenztriazole (HOBT) and EDC (0.245 g; 1.28 mol) and stirried
at RT overnight. The reaction mixture was poured into 5.0 mL of water
and the organic layer was separated. The aqueous layer was back
extracted with 5 mL of dichloromethane. The combined organics were
10 washed with 5.0 mL of 0.5N aqueous hydrochloric acid, 5 mL of
saturated aqueous sodium bicarbonate solution dried over anhydrous
magnesium sulfate, and concentrated to yield 0.751 g of the crude
product as a yellow foam. A solution of this crude product in
dichloromethane was chromatographed on 25 g of silica gel and eluted
15 first with hexanes/acetone/dichloromethane (70/25/5) and then with
hexanes/acetone/dichloromethane (65/30/5). This gave 0.63 g of the title
compound as a white solid. 1H NMR (CDCl3, 400MHz) Compound
exists as a 3:2 mixture of rotamers o 7.40-7.10 (m, 6H), 7.06 (d, 1/3H),
7.02 (t, 1/3H), 6.90 (t, 1/3H), 6.55 (d, 1/3H), 5.15 (m, lH), 4.95 (bs, lH),
20 4.63 (bd, 1/3H), 4.57-4.40 (m, 2 2/3 H), 4.10 (bd, 1/3H), 4.00 (bd, 1/3H),
3.82 (t, lH), 3.78-3.62 (m, 2H), 3.60-3.50 (m, lH), 3.04 (q, lH), 2.87 (s,
lH), 2.86 (s, 2H), 2.80-2.60 (m, lH), 1.90 (bs, lH), 2.85-2.75 (m, lH),
1.82-1.60 (m, 3H), 1.55-1.45 (m, lH), 1.45 (s, 4H), 1.42 (s, 2H), 1.39 (s,
9H).
Step C: N-[l(R)-[(1,2-Dihydro-1-methanesulfonylspiro[3H-indole-
3,4'-piperidin]- 1 '-yl)carbonyl]-2-(phenylmethyloxy)ethyl]-2-
amino-2-methylpropanamide hydrochloride
To 0.637 g (0.101 mmol) of the intermediate from Step B in
30 5 mL of dichloromethane was added 2.5 mL of trifluoroacetic acid and
stirred at RT for 30 min. The reaction rnixture was concentrated to an oil,
taken up in 10 mL of ethyl acetate and washed with 8 mL of 10~o
aqueous sodium carbonate solution. The aqueous layer was further
extracted with 5 mL of ethyl acetate. The combined organics were
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washed with 10 mL of water, dried over magnesium sulfate, filtered and
concentrated to give the 0.512 g of the free base as a white foam.
To 0.512 g of the free base in 5 mL of ethyl acetate at 0~C
was added 0.2 mL of saturated hydrochloric acid in ethyl acetate and
5 stirred for l.S h. The white precipitate was filtered under nitrogen,
washed with ether, and dried to give 0.50 g of the title compound as a
white solid lH NMR (400MHz, CD30D) Compound exists as 3:2
mixture of rotamers. o 7.40-7.28 (m, 4H), 7.25-7.17 (m, 2H), 7.08 (t,
1/3H), 7.00 (t, 1/3H), 6.80 (d, 1/3H), 5.16 (ddd, lH), 4.60-4.42 (m, 3H),
10 4.05 (t, lH), 3.90 (bs, 2H), 3.83-3.70 (m, 2H), 3.30-3.15 (m, lH0, 2.97 (s,
lH), 2.95 (s, 2H), 2.90-2.78 (m, lH), 1.96 (t, 1/3H), 1.85-1.65 (m, 4H),
1.63 (s, 2H), 1.60 (s, 4H).
EXAMPLE 3 (METHOD 2)
N-~ 1 (R)-l(1,2-Dihydro- 1 -methanesulfonylspiro[3H-indole-3,4'-piperdin] -
1'-yl) carbonyl]-2-(phenylmethyloxy)ethyl]-2-amino-2-
methylpropanamide
~0 Step A: (2R)-[[[-2-(1,1-dimethylethoxy)carbonyl]amino]-2,2-
dimethyl- 1 -oxoethyl]amino-2-(phenylmethoxy)ethyl]- 1 -
propanoic acid allyl ester
Prepared from (2R)-2-[(1,1-dimethylethoxy)carbonyl]-
amino-3-(phenylmethyloxy)ethyl-propanoic acid and allyl alcohol by
25 carrying out the coupling reaction in CH2Cl2 in the presence of EDC and
DMAP. lH NMR (400MHz, CDCl3) a 7.25 (s, SH), 5.8 (m, lH), 5.2
(dd, 2H), 5.0 (bs, lH), 4.7 (m, lH), 4.6 (m, 2H), 4.4 (dd, 2H), 3.9 (dd,
lH), 3.6 (dd, lH), 1.45 (d, 6H), 1.39 (s, 9H).
~0 Step B: (2R)-[[[-2-(1,1-dimethylethoxy)carbonyl]amino]-2,2-
dimethyl- 1 -oxoethyl]amino-2-(phenylmethyloxy)ethyl)- 1 -
propanoic acid
To a stirred solution of the crude inteImediate obtained in
Step A (6.7 g, 15.9 mmol), tetrakis (triphenylphosphine)-palladium (1.8
35 g, 0.1 eq) and, triphenyl phosphine (1.25 g, 0.3 eq) was added a solution
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of potassium-2-ethyl hexanoate (35 mL, 0.5M solution in EtOAc). The
reaction mixture was stirred at room temperature under nitrogen
atmosphere for lh and then diluted with ether (100 mL) and poured into
ice-water. The organic layer was seperated and the aqueous fraction was
5 acidified with citric acid (20%), then extracted with EtOAc. The EtOAc
extracts were washed with brine, dried over magnesium sulfate, filtered
and evaporated to give the title compound as a solid. ] H NMR (400Hz,
CD30D) o 7.3 (s, 5H), 4.7 (m, lH), 4.5 (s, 2H), 4.0 (m, lH), 3.6 (m, lH),
1.4 (d, 6H), 1.3 (s, 9H).
Step C: N- [1 (R)- [(1,2-Dihydro- 1 -methanesulfonylspiro~3H-indole-
3,4'-piperdin]- 1 '-yl)carbonyl]-2-(phenylmethyloxy)ethyl]-2-
[(1,1 -dimethyl-ethoxy)carbonyl]amino-2-methyl-
propanamide
To a solution of 1.0 g (3.44 mmol) of l-methanesulfonyl-
spiro[indoline-3,4'-piperidine] hydrochloride, 1.44 g (3.78 mmol) of
(2R)-[[-2-(1,1 -dimethylethoxy)carbonyl)amino]-2,2-dimethyl- 1 -oxo-
ethyl]-amino-2-(phenylmethyloxy)ethyl)-1-propanoic acid, N-methyl
morpholine (0.58 mL; 5.20 mmol), and l-hydroxybenztriazole (HOBT)
20 (0.58 g; 3.78 mmol), in 50 mL of dichloromethane was added EDC (1.03
g; 5.20 mmol) and stirred at RT for 16h. The reaction mixture was
diluted with an additiona~ 50 mL of dichloromethane and washed with
aqueous sodium bicarbonate solution (50 mL), dried over anhydrous
m~nesium sulfate, filtered, and concentrated. Flash chromatography (50
25 g silica gel) of the crude oily residue gave 2.148 g (90%) of the desired
material as a colorless foam. lH NMR (CDC13, 400MHz) Compound
exists as a 3:2 mixture of rotamers o 7.40-7.10 (m, 6H), 7.06 (d, 1/3H),
7.02 (t, 1/3H), 6.90 (t, 1/3H), 6.55 (d, 1/3H), 5.15 (m, lH), 4.95 (bs, lH),
4.63 (bd, 1/3H), 4.57-4.40 (m, 2 2/3 H), 4.10 (bd, 1/3H), 4.00 (bd, 1/3H),
30 3.82 (t, lH), 3.78-3.62 (m, 2H), 3.60-3.50 (m, lH), 3.04 (q, lH), 2.87 (s,
lH), 2.86 (s, 2H), 2.80-2.60 (m, lH), 1.90 (bs, lH), 2.85-2.75 (m, lH),
1.82-1.60 (m, 3H), 1.55-1.45 (m, lH), 1.45 (s, 4H), 1.42 (s, 2H), 1.39 (s,
9H).
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Step D: N-[l(R)-[(1,2-Dihydro-l-methanesulfonylspiro[3H-indole-
3,4'-piperdin]- 1 '-yl)carbonyl]-2-(phenylmethyloxy)ethyl]-2-
amino-2-methylpropanamide hydrochloride
To a solution of 2.148 g (3.41 mmol) of the intermediate
S from Step C in 10 mL of dichloromethane was added 5 mL of
trifluoroacetic acid and stirred for lh. The reaction mixture was
concentrated and basified with 100 mL of 5% aqueous sodium carbonate
solution and extracted with dichloromethane (3X50 mL). The combined
organics were washed with brine (50 mL), dried over anhydrous
10 potassium carbonate, filtered, and concentrated to yield a colorless foam.
To a solution of the foam in 25 mL of ethyl acetate at 0~C was added 4
mL of lM solution of hydrochloric acid in ethyl acetate. The precipitate
was filtered and washed ~lrst with ethyl acetate and then with ethyl
acetate-ether (1: 1), dried to yield 1.79 g (93%) of the title compound as a
15 colorless solid. lH NMR (400MHz, CD30D) Compound exists as 3:2
mixture of rotamers. o 7.40-7.28 (m, 4H), 7.25-7.17 (m, 2H), 7.08 (t,
1/3H), 7.00 (t, 1/3H), 6.80 (d, 1/3H), 5.16 (ddd, lH), 4.60-4.42 (m, 3H),
4.05 (t, lH), 3.90 (bs, 2H), 3.83-3.70 (m, 2H), 3.30-3.15 (m, lH0, 2.97 (s,
lH), 2.95 (s, 2H), 2.90-2.78 (m, lH), 1.96 (t, 1/3H), 1.85-1.65 (m, 4H),
1.63 (s, 2H), 1.60 (s, 4H).
EXAMPLE 4
N- [1 (R)- [(1,2-Dihydro- 1 -methanesulfonylspiro[3H-indole-3,4'-piperdin] -
25 1'-yl)carbonyl]-2-(phenylmethyloxy)ethyl]-2-amino-2-methyl-
prop~n~mide methanesulfonate
This compound was prepared by the treating the free base
obtained in Example 4, Step D, with methane sulfonic acid. The title
compound was obtained by recryst~lli7ing it from ethyl acetate-ethanol-
30 water. m.p. = 166~-168~C.
. .
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EXAMPLE 5
Procedure for Manufacturing Tablets of 5.0 mg Potency Active
Ingredient
s
Ingredient Per Tablet Per 25.000Tablets
Active Ingredient 5.91 mg 147.8 g
(N- [1 (R)- [(1,2-dihydro- 1 -methane-
sulfonylspiro[3H-indole-3,4'-piperdin]-
1 '-yl)carbonyl]-2-(phenylmethyloxy)-
ethyl] -2-arnino-2-methylpropanamide
methanesulfonate)
Calcium Phosphate Dibasic 188.10 mg 4.70 kg
Starch PregelAtini~e~l 120.00 mg 3.00 kg
NF 1500
Microcrystalline Cellulose NF 60.00 mg 1.50 kg
Avicel PH 101
Magnesium Stearate Impalpable 2.00 mg 50.0 g
Powder NF
Croscarmellose Sodium NF 24.00 mg 600 g
Ethanol 95% 30 ,ul 750 ml
Water purified 90 ,ul 2.25 1
(Tablet Weight = 400 g)
The active ingredient (equivalent to 5.0 mg anhydrous free
base per tablet) was mixed with the calcium phosphate dibasic, the starch
pregelatinized NF 1000, the microcrystalline cellulose NF, and half of the
30 croscarmellose sodium NF in a high Fielder 10/25 mixer for about 6
minutes. The 25% ethanol/water granulating solution was slowly added
to the powder mixture with the mixer r~lnning over a period of about 1.5
minutes then granulated for about 8 minutes to form granules. The wet
granules were dried at about 47~C (range 46 to 48~C) in a tray dryer or a
35 fluid bed dryer for approximately 3.0 hours. The dried granules were
then milled using a Quadro Comill to achieve fine granules. After
millin~, the remainder of the croscarmellose sodium NFS was added to
the fine granules and mixed in a V blender for about 10 minutes.
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Magnesium stearate imp~lr)able powder NF was added to this blend
through a 60 mesh stainless steel screen and blended in the V blender for
about 1 minute. The lubricated mixture was compressed to provide
tablets of 5.0 mg active ingredient (free base equivalent).
s
EXAMPLE 6
Procedure for Manufacturing Coated Tablets of 5.0 mg Potency Active
In~redient
In~redient Per Tablet Per 25~000 Tablets
Hydroxypropyl Methylcellulose 3.2 mg 80 g
USP (HPMC)
Hydroxypropyl Cellulose NF 3.2 mg 80.0 g
with < 0.3% Silica (HPC)
Titanium Dioxide USP 1.28 mg 32.0 g
Talc USP Purified 0.32 mg 8.0 g
Water Purified To 80 111 To 200 ml
(Film Coated Tablet Weight = 408 g)
The titanium dioxide and talc, USP were mixed and passed
through a 60 mesh stainless steel screen. This mixture was mixed with
HPMC and HPC to form a dry blend. The dry blend was added to water
(20 ml) which was previously heated to 90~C with mild agitation to
25 ensure that the blend is wetted to form a slurry. The rem~in(ler of the
water (up to 32 ml) was added to the slurry at ambient temperature with
gentle agitation to form a suspension. The suspension was then applied
to the tablets from the previous Example using the following guidelines
to provide the coated tablets.
30 Pan: suitable size
Pan Speed: 20 RPM
Nozzles: 2850 liquid/120 air
Inlet Temperature: 85~C
Bed Temperature: 47~C
35 Spray Rate: ca. 2.0 g/minute/kg Tablets
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EXAMPLE 7
Procedure for Manufacturing Tablets of 25 mg Potency Active
Ingredient
s
In~redient Per Tablet Per 25~000Tablets
ActiveIngredient 29.55 mg 738.75 g
(N-[1 (R)-[(1,2-dihydro- 1 -methane-
sulfonylspiro[3H-indole-3,4'-piperdin]-
l '-yl)carbonyl]-2-(phenylmethyloxy)-
ethyl]-2-amino-2-methylprop~n~mide
methanesulfonate)
Calcium Phosphate Dibasic 174.46 mg 4.361 kg
Starch Pregel~tini7e~1 113.00 mg 2.825 kg
NF 1500
Microcrystalline Cellulose NF 57.00 mg 1.425 kg
Avicel P~I 101
Magnesium Stearate Impalpable 2.00 mg 50.0 g
Powder NF
Croscarmellose Sodium NF 24.00 mg 600 g
Ethanol 95~o 30 ~11 750 ml
Water purified 90 ~1 2.25 1
(Tablet Weight = 400 g)
The active ingredient (equivalent to 25 mg anhydrous free
base per tablet) was mixed with the calcium phosphate dibasic, the starch
35 pregel~tini~e.d NF 1000, the microcrystalline cellulose NF, and half of the
croscarmellose sodium NF in a high shear granulator Fielder 10/25 mixer
for about 6 minutes. The 25% ethanoltwater granulating solution was
slowly added to the powder mixture with the mixer rlmning over a period
of about 1.5 minutes then granulated for about 8 minutes to form
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granules. The wet granules were dried at about 47~C (range 46 to 48~C)
in a tray dryer or a fluid bed dryer for approximately 3.0 hours. The dried
granules were then milled using a Quadro Comill to achieve fine
granules. After milling, the remainder of the croscarmellose sodium NFS
5 was added to the fine granules and mixed in a V blender for about 10
minutes. Magnesium stearate impalpable powder NF was added to this
blend through a 60 mesh stainless steel screen and blended in the V
blender for about 1 minute. The lubricated mixture was compressed to
provide tablets of 25 mg active ingredient (free base equivalent).
EXAMPLE 8
Procedure for Manufacturing Coated Tablets of 25 mg Potency Active
Ingredient
Ingredient Per Tablet Per 25,000 Tablets
Hydroxypropyl Methylcellulose 3.2 mg 80 g
USP (HPMC)
Hydroxypropyl Cellulose NF 3.2 mg 80.0 g
with < 0.3~o Silica (HPC)
Titanium Dioxide USP 1.28 mg 32.0 g
Talc USP Purified 0.32 mg 8.0 g
Water Purified To 80 ~1 To 200 ml
(Film Coated Tablet Weight - 408 g)
Using essentially the procedure of Example 9 and applying
the suspension to the tablets from the previous Example, 25 mg potency
coated tablets were formed.
. . " . .
CA 022~3390 1998-10-29
WO 97/41879 PCT/US97/07516
EXAMPLE 12
Clinical Study of N-[l(R)-[(1,2-Dihydro-1-methane-sulfonylspiro[3H-
indole-3 ,4'-piperidin]- 1 '-yl)carbonyl]-2-(phenyl-methyloxy)ethyl]-2-
5 amino-2-methylpropanamide methanesulfonate in Healthy Youny Adults
In this study, 9 healthy young men (ages 18 to 30 years) who
did not suffer sleep complaints were randomly assigned to a sequence of
3 treatment periods. In each period the subjects received a single oral
dose of either placebo, 5 mg of N-~l(R)-[(1,2-dihydro-1-methane-
10 sulfonylspiro[3H-indole-3,4'-piperidin]-1'-yl)carbonyl]-2-(phenylmethyl-
oxy)ethyl]-2-amino-2-methyl-propanamide methanesulfonate or 25 mg of
N-[l(R)-[(1,2-dihydro-1-methanesulfonylspiro[3H-indole-3,4'-piperidin]-
1 '-yl)carbonyl] -2-(phenylmethyloxy)ethyl]-2-amino-2-methyl-
prop~n~mide methanesulfonate once daily for 7 days. Sleep was
15 recorded for 2 nights: a habituation night and a blood sampling night
(Days 6 and 7 of study drug ~lmini.~tration). On average, the durations of
Stages 1, 2, and 3 were simi~r in the three study conditions. After 7 days
of treatment, however, Stage 4 sleep duration was significantly higher
after ~rlmini~tration of 25 mg of N-[l(R)-[(1,2-dihydro-1-methane-
20 sulfonyl-spiro[3H-indole-3,4'-piperidin]- 1 '-yl)carbonyl]-2-(phenylmethyl-
oxy)ethyl]-2-amino-2-methylpropanamide methanesulfonate than after
treatment with placebo (54 + 27 min versus 37 + 19 min, N = 8, p <
0.05). Moreover, when compared to placebo, there was a significant
increase in REM sleep in the 25 mg treatment group on the blood
25 sampling night (103 + 9 min versus 88 + 19 min, N = 9, p < 0.05) and
there was a reduction in sleep latency, a reduction in the amount of time
spent awake, as well as a decrease in the number of sleep disturbances in
the 25 mg treatment group. Accordingly, the group treated with 25 mg of
growth hormone secretagogue exhibited improved sleep efficiency and
30 sleep maintenance. This study suggests that use of a growth hormone
secretagogue may have a beneficial effect in enhancing the quality of
sleep in humans.
CA 022~3390 l998-l0-29
W O97/41879 PCT~US97/07516 -
While the invention has been described and illustrated with
reference to certain particular embodiments thereof, those skilled in the
art will appreciate that various adaptations, changes, modifications,
substitutions, deletions, or additions of procedures and protocols may be
5 made without departing from the spirit and scope of the invention. For
example, effective dosages other than the particular dosages as set forth
herein above may be applicable as a consequence of variations in the
responsiveness of the m~mm~l being treated for any of the indications
with the compounds of the invention indicated above. Likewise, the
10 specific pharmacological responses observed may vary according to and
depending upon the particular active compounds selected or whether
there are present ph~rmaceutical carriers, as well as the type of
formulation and mode of admini~tration employed, and such expected
variations or differences in the results are contemplated in accordance
15 with the objects and practices of the present invention. It is intended,
therefore, that the invention be defined by the scope of the claims which
follow and that such claims be interpreted as broadly as is reasonable.
