U.S. patent application number 10/472741 was filed with the patent office on 2004-07-08 for n-(2-arylethyl) benzylamines as antagonists of the 5-ht6 receptor.
Invention is credited to Chen, Zhaogen, Cohen, Michael Philip, Fisher, Matthew Joseph, Giethlen, Bruno, Gillig, James Ronald, McCowan, Jefferson Ray, Miller, Shawn Christopher, Schaus, John Mehnert.
Application Number | 20040132800 10/472741 |
Document ID | / |
Family ID | 26959969 |
Filed Date | 2004-07-08 |
United States Patent
Application |
20040132800 |
Kind Code |
A1 |
Chen, Zhaogen ; et
al. |
July 8, 2004 |
N-(2-arylethyl) benzylamines as antagonists of the 5-ht6
receptor
Abstract
The present invention provides compounds of formula (I), which
are antagonists of the 5-HT.sub.6 receptor. 1
Inventors: |
Chen, Zhaogen; (Noblesville,
IN) ; Cohen, Michael Philip; (lndianapolis, IN)
; Fisher, Matthew Joseph; (Mooresville, IN) ;
Giethlen, Bruno; (Altorf, FR) ; Gillig, James
Ronald; (Indianapolis, IN) ; McCowan, Jefferson
Ray; (Indianapolis, IN) ; Miller, Shawn
Christopher; (Morgantown, IN) ; Schaus, John
Mehnert; (Zionsville, IN) |
Correspondence
Address: |
ELI LILLY AND COMPANY
PATENT DIVISION
P.O. BOX 6288
INDIANAPOLIS
IN
46206-6288
US
|
Family ID: |
26959969 |
Appl. No.: |
10/472741 |
Filed: |
February 27, 2004 |
PCT Filed: |
March 15, 2002 |
PCT NO: |
PCT/US02/05115 |
Current U.S.
Class: |
514/419 ;
548/503 |
Current CPC
Class: |
C07C 225/16 20130101;
A61P 25/22 20180101; A61P 25/28 20180101; C07D 213/74 20130101;
C07D 333/20 20130101; C07F 7/0812 20130101; C07D 213/65 20130101;
A61P 25/00 20180101; C07C 323/32 20130101; C07D 233/24 20130101;
C07D 307/91 20130101; C07D 417/12 20130101; C07C 211/52 20130101;
C07C 211/56 20130101; C07C 217/60 20130101; C07D 403/06 20130101;
C07D 209/16 20130101; C07D 403/12 20130101; C07D 405/12 20130101;
C07D 277/34 20130101; C07D 213/38 20130101; A61P 25/18 20180101;
C07C 217/58 20130101; C07D 209/14 20130101; C07D 401/12 20130101;
C07D 209/08 20130101; C07D 239/34 20130101; A61P 43/00 20180101;
C07C 311/37 20130101; C07D 213/68 20130101; C07D 209/86 20130101;
C07C 317/34 20130101; C07D 471/04 20130101; C07D 213/64
20130101 |
Class at
Publication: |
514/419 ;
548/503 |
International
Class: |
C07D 209/04; A61K
031/404 |
Goverment Interests
[0001] This invention was discovered in the performance of U.S.
government supported research under grants NIMH KO2MH01366 and
NCEMS GM52213, and the U.S. government may have certain rights in
the invention.
Claims
Having thus described our invention, what we now claim as new and
desire to secure by Letters Patent is as follows:
1. A compound having the formula: 7wherein, R.sub.1 is selected
from the group consisting of hydrogen, methyl and other lower
alkyls and may be different at each location, R.sub.2 is selected
from the group consisting of hydrogen, a methyl, ethyl, n-propyl,
and phenyl addition, and R.sub.3 is selected from the group
consisting of hydrogen, methyl, methoxy and substituted and
unsubstituted lower alkyls.
2. The compound of claim 1 wherein R.sub.2 is an ethyl group.
3. A compound having the formula: 8wherein, R.sub.1 is selected
from the group consisting of hydrogen, and methyl and other lower
alkyls and may be the same or different at each site; and R.sub.4
is selected from the group consisting of hydrogen, lower alkyls,
aryls, substituted and unsubstituted heteroaryls; and R.sub.3 is
selected from the group consisting of hydrogen, methyl and methoxy
and substituted and unsubstituted lower alkyls.
4. The compound of claim 4 where R.sub.4 is selected from the group
consisting of CH.sub.2-phenyl, H, and SO.sub.2-phenyl.
5. A compound having the formula: 9wherein, R.sub.1 is selected
from the group consisting of hydrogen, methyl and other lower
alkyls and may be the same or different at each site; R.sub.3 is
selected from the group consisting of hydrogen, methyl and methoxy
and substituted and unsubstituted lower alkyls, and X is selected
from the group consisting of oxygen and 2H.
6. A therapeutically effective composition for treating a condition
mediated by the 5-HT.sub.6 receptor, comprising: a pharmaceutically
acceptable carrier; and a compound selected from the group
consisting of: 10wherein X is selected from the group consisting of
oxygen and 2H; wherein R.sub.1 is selected from the group
consisting of hydrogen, methyl and other lower alkyls and may be
the same or different at each site; wherein R.sub.3 is selected
from the group consisting of hydrogen, methyl and methoxy and
substituted and unsubstitued lower alkyls; and wherein each of
R.sub.2 and R.sub.4 are selected from the group consisting of
hydrogen, lower alkyls, aryls, alkaryls, substituted aryls, and
heteroaryls.
7. The composition of claim 6 wherein R.sub.2 and R.sub.4 are
selected from the group consisting of CH.sub.2-phenyl,
SO.sub.2-phenyl, and hydrogen.
8. The composition of claim 6 wherein the R.sub.3 moiety is in the
2 position.
9. A method for treating a condition mediated by the 5-HT.sub.6
receptor, by administering an effective amount of a therapeutically
effective composition comprising: a pharmaceutically acceptable
carrier; and a compound selected from the group consisting of:
11wherein X is selected from the group consisting of oxygen and 2H;
wherein R.sub.1 is selected from the group consisting of hydrogen,
methyl and other lower alkyls and may be the same or different at
each site; wherein R.sub.3 is selected from the group consisting of
hydrogen, methyl and methoxy and substituted and unsubstituted
lower alkyls; and wherein each of R.sub.2 and R.sub.4 are selected
from the group consisting of hydrogen, lower alkyls, aryls,
alkaryls, substituted aryls, and heteroaryls.
10. A method for testing antagonists and antagonists with
selectivity for the 5-HT.sub.6 receptor comprising: administering a
compound selected from the group consisting of: 12wherein X is
selected from the group consisting of oxygen and 2H; wherein
R.sub.1 is selected from the group consisting of hydrogen, methyl
and other lower alkyls and may be the same or different at each
site; wherein R.sub.3 is selected from the group consisting of
hydrogen, methyl and methoxy and substituted and unsubstituted
lower alkyls; and wherein each of R.sub.2 and R.sub.4 are selected
from the group consisting of hydrogen, lower alkyls, aryls,
alkaryls, substituted aryls, and heteroaryls; and observing said
animals' responses; and comparing said responses to control
animals; and administering other compounds of unknown activity to
said experimental animals.
11. The compound of claim 1 further comprising a radiolabel.
12. The compound of claim 3 further comprising a radiolabel.
13. The compound of claim 5 further comprising a radiolabel.
Description
DESCRIPTION
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] The present invention relates to the synthesis of novel
ligands selective for a subgroup of receptors for serotonin (5-HT).
While there are seven subgroups of 5-HT receptors, this invention
is selective for the 5-HT.sub.6 subgroup. This invention also
relates to the synthesis of novel ligands selective for the
5-HT.sub.6 subgroup receptor that act as agonists to the natural
ligands for this receptor. The invention also relates to the
creation of novel ligands that act as antagonists to the 5-HT.sub.6
receptor. The invention further relates to use of said compounds to
treat mammals adversely affected by conditions mediated by the
5-HT.sub.6 receptor.
[0004] 2. Description of the Prior Art
[0005] Serotonin receptors have been divided into a number of
families and subfamilies (5-HT.sub.1-5-HT.sub.7) and approximately
14 populations have been cloned. One of the newest populations
identified is the 5-HT.sub.6 subgroup. It has been observed that
various tricyclic psychotropic agents (neuroleptics,
antidepressants, and atypical neuroleptics agents) bind the
5-HT.sub.6 receptor with nanomolar affinities (Roth et al. J.
Pharmacol. Exp. Ther. 1994, 268, 1403-1410). A rat 5-HT.sub.6
receptor was first cloned in 1993 and, more recently, the same
group described the cloning of a human 5-HT.sub.6 receptor. The
5-HT.sub.6 serotonin receptors are members of the G-protein
superfamily, are positively coupled to an adenylate cyclase second
messenger system, and are found primarily in the central nervous
system. Serotonin bound to the 5-HT.sub.6 receptor subgroup causes
an activation of the adenylate cyclase enzyme, with concomitant
increased levels of intracellular cAMP. Although the exact
physiological function and clinical significance of the 5-HT.sub.6
receptor subgroup is not known, as noted above, many anti-psychotic
agents bind these receptors with high affinity. Also, in rats that
do not express 5-HT.sub.6 receptors, the animals behave in a manner
that seems to involve an increase in cholinergic function,
suggesting that 5-HT.sub.6 specific ligands might be of value in
the treatment of anxiety-related disorders and memory deficits.
[0006] Upon binding to cellular receptors, ligands may act as
agonists or antagonists to endogenous receptor-ligand function. In
the case of the 5-HT.sub.6 receptor, several specific ligands have
been discovered which act as 5-HT.sub.6 specific antagonists, but
prior to the present invention, selective ligands which act as
agonists to the 5-HT.sub.6 receptor were unknown.
SUMMARY OF THE INVENTION
[0007] It is an object of the invention to create derivatives of
serotonin (5-HT) that specifically bind the 5-HT.sub.6 receptor
subgroup of the serotonin receptor family. It is another object of
this invention to create 5-HT.sub.6-selective ligands that act as
agonists when bound to the 5-HT.sub.6 receptor. It is further an
object of this invention to create 5-HT.sub.6-selective ligands
that act as antagonists when bound to the 5-HT.sub.6 receptor.
Furthermore, the compounds of the present invention that possess
antagonist activity are tryptamine derivatives and are structurally
unrelated to previously described 5-HT.sub.6 antagonists. It is
further an object of this invention to administer 5-HT.sub.6
selective ligands to animals to determine the physiological and
biochemical effects of specific activation and inhibition of
5-HT.sub.6 receptor function. Finally, it is an object of this
invention to treat mental disorders mediated by 5-HT.sub.6 function
by administering to treatment subjects the 5-HT.sub.6-selective
agonists and antagonist compounds described herein.
[0008] Various indolealkylamines, including serotonin(5-HT) and
5-methoxytryptamine, have been observed to bind the 5-HT.sub.6
receptor with high affinity and produce a potent dose-dependent
increase in cAMP levels. These tryptamines, however, are
non-selective and bind at multiple families of 5-HT receptors.
According to the invention, various modifications of 5-HT have been
made to generate ligands with selectivity for the 5-HT.sub.6
receptor. An analog of 5-HT with a 2-methyl substituent introduced
(2-methyly-5-HT) binds the 5-HT.sub.6 receptor with an affinity
equivalent to that of the parent compound. The above analog is
selective for the 5-HT.sub.6 and 5-HT.sub.3 receptors and binds at
the 5-HT.sub.6 subgroup with a 20 fold greater affinity than at
5-HT.sub.3 receptors.
[0009] A 2-methyl analog of 5-methoxytryptamine,
5-methoxy-2-methyltryptam- ine, binds to the 5-HT.sub.6 receptor
with an affinity comparable to 2-methyl-5-HT. However,
5-methoxy-2-methyltryptamine lacks affinity for 5-HT.sub.3
receptors. Thus, 5-methoxy -2-methyltryptamine presents a ligand
with specificity for the 5-HT.sub.6 receptor subgroup. In the
present invention, the 5-methoxy-2-methyltryptamine compound has
been modified and several of its alkyl derivatives bind with
comparable affinity and activate adenylate cyclase activity at
levels comparable to serotonin. Furthermore, one derivative,
5-methoxy-2-phenylotryptamine, binds to the 5-HT.sub.6 receptor
with a high affinity but the phenyl addition renders the compound
an antagonist to 5-HT stimulated adenylate cyclase activity.
BRIEF DESCRIPTION OF THE DRAWINGS
[0010] The foregoing and other objects, aspects and advantages will
be better understood from the following detailed description of the
preferred embodiments of the invention with reference to the
drawings, in which:
[0011] FIG. 1 is a graph showing the adenylate cyclase activity
observed with several compounds of the invention.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS OF THE
INVENTION
[0012] The invention provides novel tryptamine derivative compounds
with selectivity for the 5-HT.sub.6 receptor subgroup. For the
present purpose, an agent is termed selective when it displays an
affinity for 5-HT.sub.6 receptors that is tenfold higher than
affinities it displays for other related receptor populations. The
invention further provides a method using said compounds to
receptor subtype 5-HT.sub.6 as agonists, or as antagonists to
serotonin. The compounds of the invention can be used either as the
free base or as the pharmaceutically acceptable acid-addition salt
form, for example, hydrochloride, hydrobromide, tartrate, and
maleate. They may be used in oral or injectable pharmaceutical
preparations as prophylactic and acute-phase remedies for the
relief and reversal of serotonin-regulated symptoms. They may be
used alone or in combination with each other or other known
medications. Finally, said compounds may be used as above for
determining 5-HT.sub.6 receptor function.
[0013] Serotonin (5-hydroxy tryptamine, or 5-HT) is a product of
tryptophan metabolism and is a tryptamine derivative that is a
potent neurotransmitter. Serotonin is a well-characterized
tryptamine derivative which regulates calcium ion channels on the
surface of nerve and muscle cells. Many mental disorders in humans
are associated with fluctuations in serotonin levels and are
effectively treated with drugs which specifically interact with
serotonin receptors or that block the reuptake of serotonin into
the presynaptic axon terminals, suggesting that serotonin
dysregulation may be involved in various mental disorders. Some
serotonin receptor ligands and are clinically approved as drugs for
the treatment of migraine headaches, depression, high blood
pressure, and psychosis.
[0014] Generally, tryptamine derivatives are non-selective and bind
at multiple 5-HT receptor subgroups. Serotonin is no exception and
binds at the various subfamilies of the 5-HT receptor, including
the 5-HT.sub.6 subgroup where it is a potent activator of adenylate
cyclase enzyme activity. Serotonin has the chemical formula: 2
[0015] Some modifications of serotonin result in loss of affinity
for various 5-HT receptor subgroups. It had previously been thought
that introduction of a 2-methyl substituent to 5-HT was not
tolerated by any 5-HT receptors but the 5-HT.sub.3 subgroup. Thus,
prior to identification of the 5-HT.sub.6 receptor, 2-.degree.
methylation of 5-HT was thought to render the product selective for
the 5-HT.sub.3 subgroup. We have found that the two methyl
derivative of 5-HT, 2-methyl-5-HT has a high affinity for the
5-HT.sub.6-receptor. In fact, 2-methyl-5-HT binds the 5-HT.sub.6
receptor with a 20 fold greater affinity over 5-HT.sub.3 receptors.
The 5-HT.sub.6-selective ligand 2-methyl-5-methoxytryptamine
contains a primary amine, presenting an obstacle to the compound
crossing the blood brain barrier and also rendering the compound
vulnerable to rapid metabolism due to oxidative deamination. Our
discovery that a methyl substituent at the 2 position was tolerated
by the 5-HT.sub.6 receptor, together with the previous observation
that O-.degree. methylation of 5-HT abolishes affinity for
5-HT.sub.3 receptor, led to the present invention.
[0016] To address the above limitations of
2-methyl-5-methoxytryptamine, several derivative compounds were
synthesized that were lipophilic and also might be less prone to
rapid metabolism. N,N-dimethyl substituents were added to
2-methyltryptamine to create 2-methyl-N,N-dimethyltryptamin- e
(Ki=308 nM). Re-introduction of the methoxy group to this compound,
to form 2-methyl-5-methoxy-N,N-dimethyltryptamine resulted in a
compound (Compound A) with an affinity for the 5-HT.sub.6 receptor
of Ki=60 nM. Homologation of the 2-methyl substituent of the above
compound to form 2-ethyl-5-methoxy-N,N-dimethyltryptamine resulted
in a ligand with an increased affinity for the 5-HT.sub.6 of Ki=16
nM (Compound B). To determine whether or not greater bulk additions
could be added in place of a methyl or ethyl group, the 2-phenol
derivative of the 2-methyl-5-methoxy-N,N-dimethyltryptamine was
generated. This compound D binds the 5-HT.sub.6 receptor with a
Ki=20 nM. These derivatives were of the general formula 1 3
[0017] where R.sub.1 and R.sub.2=H or CH.sub.3,
[0018] R.sub.3=H, OH, OCH.sub.3, or a substituted or unsubstituted
alkyl,
[0019] R.sub.4=H, CH.sub.2-phenyl, SO.sub.2-phenyl, or CH.sub.2 as
part of a substituted or unsubstituted alkyl ring connecting
R.sub.4 and R.sub.5,
[0020] R.sub.5=H, CH.sub.3, or CH.sub.2 as part of a substituted or
unsubstituted alkyl ring connecting R.sub.5 with either R.sub.4 or
R.sub.6,
[0021] R.sub.6=H, CH.sub.3, or CH.sub.2 as part of a substituted or
unsubstituted alkyl ring connecting R.sub.6 and R.sub.5.
[0022] The compounds of the invention and the pharmaceutically
acceptable salts of the compounds of the invention can be used in
the form of pharmaceutical preparations. The preparations can be
administered orally, for example in the form of tablets, coated
tablets, dragees, hard and soft gelatin capsules, solutions,
emulsions or suspensions. The administration can be effected
rectally, for example in the form of suppositories, or
parenterally, for example in the form of injection solutions.
[0023] The compounds of the invention can be processed with
pharmaceutically inert, inorganic or organic carriers for the
production of pharmaceutical and research preparations. The
preparations can contain preservatives, solubilizers, wetting
agents, emulsifiers, sweeteners, colorants, flavorants, salts for
varying the osmotic pressure, buffers, masking agents or
antioxidants. They can also contain still other therapeutically
valuable substances.
[0024] The compounds of the present invention can also be
radiolabelled and used to identify other 5-HT.sub.6 ligands using
techniques common in the art. This can be achieved by incubating
the receptor in the presence of a ligand candidate plus an
equimolar amount of radiolabelled compound of the invention.
Ligands selective for 5-HT.sub.6 are then revealed as those that
are not significantly displaced by the compounds of the present
invention.
[0025] Another embodiment of the invention can be the
administration of the compounds of the invention to animals in drug
discrimination assays. In a drug discrimination paradigm, animals
(usually rats) can be trained to recognize the effects of a given
agent. Once trained, these animals can be used in tests of stimulus
generalization to identify other agents that produce similar
effects (i.e., agonists), or the animals can be used in tests of
stimulus antagonism to identify agents that block or antagonize
(i.e., antagonists) the effects of the training drug. Hence, the
procedure can be used to identify agonists that produce an effect
common to the training drug, more antagonists that can block the
effects of the training drug. Specifically, with a
5-HT.sub.6-selective agonist as training drug, the animals can be
used to identify other 5-HT.sub.6 agonists and to identify
5-HT.sub.6-antagonists.
[0026] One family of compounds contemplated for use in this
invention is represented by the formula 2 4
[0027] wherein R.sub.2 is selected from the group consisting of
small alkyls (e.g., methyl, ethyl, n-propryl) aryl (e.g. phenyl)
and arylalkyls.
[0028] Another family of compounds for use in this invention is
represented by formula 3 5
[0029] wherein R.sub.1 is selected from the group consisting of
lower alkyls such as ethyl and propyl, methyl and hydrogen, and can
be the same or different at each location, and R.sub.4 is from the
group comprising H, CH.sub.2-phenyl, or SO.sub.2-phenyl.
[0030] Another family of compounds for use in this invention is
represented by formula 4 6
[0031] wherein X is selected from the group consisting of oxygen or
2H and R.sub.1 is H or lower alkyls such as methyl, ethyl, or
propyl.
[0032] The following examples illustrate the present invention in
more detail. However, they are not intended to limit its scope in
any manner.
EXAMPLE 1
Synthesis of 2-ethyl-5-methoxy-N,N-dimethyltryptamine Maleate
(Compound B)
[0033] A 2.5M solution of nBuLi (1.75 ml, 4.38 mmol) was added in a
drop wise manner to a stirred solution of
2-methyl-5-methoxy-N,N-dimethyltrypt- amine (compound A in the
examples below) free base (1.00 g, 4.33 mmol) in dry THF (7 mL) at
-78.degree. C. under N.sub.2.
[0034] After stirring the reaction mixture for five minutes, the
cooling bath was removed and CO.sub.2 gas was passed into the
solution for 10 minutes. The solvent was removed at 0.degree. C.
under reduced pressure to give a transparent solid. The flask was
flushed with N.sub.2 and dry THF (7 mL) was added. The reaction
mixture was degassed at -150.degree. C. under reduced pressure of 1
mMHg, then allowed to warm to -78.degree. C.; 1.7M tNuLi (2.8 mL,
4.8 mmol) was added in a drop wise manner. The solution was kept at
-78.degree. C. for three hours. The reaction make sure was
acidified with a saturated ethereal solution of HCl. Anhydrous
Et.sub.2O was added to the resulting suspension and the supernatant
was decanted. The residue was heated at 100.degree. C. under
reduced pressure for 20 minutes. The resulting residue was purified
by flash chromatography on silica gel (CH.sub.2Cl.sub.2/MeOH; 12:1)
to give 0.17 g of a bright yellow oil (16%).
.sup.1H-NMR(CDCl.sub.3) d 8.06 (s, 1H, J=8.67 HZ), 6.98 (s, 1H),
6.76 (dd, 1H, J=2.34, 8.73 HZ), 3.84 (s, 3H) 2.91-2.87 (m, 2H),
2.71 (q, 2H, J=7.38 HZ), 2.57-2.52 (m, 2H) 2.38 (s, 6H)1.25 (t, 3H,
J=7.38 HZ).The maleate salt was prepared and recrystallized from an
EtOAc/Et.sub.2O mixture; mp 123.degree. C.
EXAMPLE 2
[0035] Magnesium turnings and NH.sub.4Cl were added to a solution
of Example 11 infra
(1-Benzenesulfonyl-5-methoxy-2-n-propyl-N,N-dimethyl tryptamine,
free base; 259 mg, 0.65 mmol) in MeOH (17 mL) and the mixture was
allowed to stir at room temperature for one-hour. Saturated
NH.sub.4Cl solution was added and the reaction mixture was
extracted with CH.sub.2Cl.sub.2. The organic portion was dried
(MgSO.sub.4) and the solvents was removed under reduced pressure.
The residue was purified by flash chromatography on silica gel
(CH.sub.2Cl.sub.2/MeOH; 9:1) to give 75 mg of a bright yellow oil.
.sup.1H-NMR (CDCl.sub.3) .quadrature.7.71 (brs, 1H), 2.89-2.83 (m,
2H), 2.69 (t, 2H, J=7.56 Hz), 2.53-2.47 (m, 2H), 2.36 (s, 6H), 1.68
(tq, 2H, J=7.28, 7.56 HZ), 0.98 (t, 3H, J=7.28 HZ). The salt was
prepared and recrystallized from acetone; mp 146-147.degree. C.
EXAMPLE 3
5-methoxy-2-phenyl-N,N-dimethyltryptamine Oxalate (Compound D)
[0036] 5-methoxy-2-phenylindole (3 g, 13.44 mmol) was added to a
stirred ice-cooled solution of 1-dimethylamino-2-nitroethylene
(1.56 g, 13.44 mmol) in trifluoracetic acid (8 ml). The resulting
mixture was allowed to stir under N.sub.2 at room temperature for
30 minutes and was then poured into ice/water. The solution was
extracted with EtOAc and the organic portion was washed
consecutively with saturated NaHCO.sub.3 solution, H.sub.20, and
then brine. The organic portion was dried (MgSO.sub.4) and solvent
was removed under reduced pressure. The residue was recrystallized
from CH.sub.2Cl.sub.2/hexane to give 2.36 g (60%) of a red powder.
.sup.1H-NMR (acetone-d6) d 8.82 (brs, 1H), 3.92 (s, 3H), IR
(Kbr)1601, 1475, 1251 cm.sup.-1. A solution of this material (2 g,
6.75 mmol) in dry THF was added in a drop wise manner to a cooled
0.degree. C. suspension of LiALH.sub.4 (1.54 g, 40.5 mmol)in dry
THF under N.sub.2. The reaction mixture was heated at reflux for
one-hour and then allowed to stand at room temperature overnight.
The resulting mixture was quenched with H.sub.2O then 15% NaOH
solution. Celite was added and the solution was filtered. The
solvent was removed under reduced pressure. The residue was
purified by flash chromatography on silica gel
(CH.sub.2Cl.sub.2/MeOH; 9:1) to give 1 g (55%) of the primary amine
as an oil. .sup.1H-NMR (CDCl.sub.3) d 8.19 (brs, 1H, J=2.37 HZ),
6.88 (dd, 1H, J=2.24, 8.75 HZ), 3.89 (s, 3H), 3.04 (brs, 4H).
IR(KBr) 3397, 3347 cm.sup.-1. Sodium cyanoborohydride (510 mg, 8.12
mmol) was added to a solution of the primary amine (700 mg, 2.63
mmol) and 37% aqueous CH.sub.2O in MeCN (10 mL) at room
temperature. The resulting mixture was adjusted to pH 5 with HOAc
and was allowed to stir at room temperature overnight. A 15%
solution of NaOH was added to neutralize the mixture and the
mixture was extracted with CH.sub.2Cl.sub.2. The combined organic
portion was washed with saturated NaHCO.sub.3 solution and brine.
The organic portion was dried (MgSO.sub.4) and solvent was removed
under reduced pressure. The residue was purified by flash
chromatography on silica gel (CH.sub.2Cl.sub.2/MeOH; 9:1) to give
195 mg of 5-methoxy-2-phenyl-dimethyltryptamine free base as a
white powder. .sup.1HNMR (CDCl.sub.3) d 8.05 (brs, 1 h), 7.56-7.53
(m, 2H),7.49-7.44 (m, 2H), 7.39-7.34 (m, 1H), 7.29, 7.25 (m, 1H),
7.11 (d, 1H, J=2.25 HZ). 6.87 (dd, 1H, J=2.52, 8.73 HZ). 3.89 (s,
3H). 3.13-3.08 (m, 2H), 2.72-2.66 (m, 2H), 2.39 (s, 6H). Although
the HCl salt has previously been reported, difficulties in his
purification led to isolation of the product has its salt; after
recrystallization from acetone.
EXAMPLE 4
4-dimethylaminomethyl-9-benzyl-6-methoxy-1,2,3,4,
Tetrahydrocarbazole Hydrochloride (Compound E)
[0037] A mixture of 4-methoxyphenyl)benzylamine (42 g 0.2 mol) and
ethyl 6-bromocyclohexanone Carboxylate (J. Org Chem 1961, 26, 22)
(24.9 g, 0.1 mol) were heated at reflux in dry benzene (250 mL) for
24 hours. The reaction mixture was cooled and precipitated
(4-methoxyphenyl) benzylamine hydrobromide was separated by
filtration. The benzene extract was concentrated and fused zinc
chloride (40 g) was added in reflux in absolute ethanol (125 mL)
for six hours. The cooled mixture was slurred in H.sub.20 (250 mL)
and extracted with (Et.sub.2O; 4.times.200 mL). The combined ether
extracts were washed with 5% HCl (100 mL), followed by brine
solution and dried with MgSO.sub.4. The ether extract was
evaporated under reduced pressure to give crude ethylester of title
compound which was treated with a solution of KOH (50 g) in
H.sub.20 (150 mL) and CH.sub.3OH (150 mL) at reflux temperature for
three hours. The solution was evaporated to dryness under reduced
pressure, the resulting residue was dissolved in H.sub.20 (250 mL)
and the aqueous solution was extracted with Et.sub.2O and acidified
with 10% HCl. The resulting solid was dried to give 22 g (33%) of
title compound and was recrystallized from 1s.PrOH-H2). Mp
212-214.degree. C. To a mixture of sodium hydride (0.96 g, 0.04
mol) in dry benzene (200 mL) was added portion wise
9-benzyl-6-methoxy-1,2,3,4-tetrahydro-4-carboxylic acid (14.00 g,
0.04 mol) and the mixture was stirred for one-hour. Thionoyl
chloride (3.00 mL, 0.04 mol) was added slowly in the stirring was
continued for 30 minutes. The resulting solution was poured into
aqueous dimethylamine solution (40%)(36.50 mL) with ice bath
cooling. The mixture was third for one-hour, washed with 100 mL
H.sub.2O, NaHCO.sub.3 (50 mL)and saturated brine solution (50 mL),
and dried with MgSO.sub.4, diluted with n-pentane (200 mL)and
cooled to give 4-dimethylaminocarbonyl-9-benzyl-6-methoxy-1,2-
,3,4-tetrahydrocarbazole (9.10 g. 60%) mp 153-155.degree. C. To a
stir it solution of LialH.sub.4 (4.71 g, 94.2 mmol) in dry THF was
added portion wise
4-dimethylaminocarbonyl-9-benzyl-6-methoxy-1,2,3,4-tetrahydrocarbazo-
le (9.00 g, 24.8 mmol) and the mixture was heated under reflux for
five hours the reaction mixture was cooled water (5.0 mL) NaOH
solution (5.0 mL) was added and filtered. The filtrate was
evaporated to dryness to give
4-dimethylamino-9-benzyl-6-methoxy-1,2,3,4-tetrahydrocarbazole (8
g, 92%). The free base was dissolved in either in converted to the
salt using ethereal hydrochloride and recrystallized from a mixture
of EtOH and Et.sub.2O. Mp 238-240.degree. C.
EXAMPLE 5
[0038] Compound F is known and was prepared according to the
following patent procedure:
4-aminomethyl-9-benzyl-1,2,3,4-tetrahydrocarbazoles, U.S. Pat. No.
3,939,177, Feb 17, 1976.
EXAMPLE 6
[0039] Compound G Sodium metal was added portion wise over a thirty
minute period to a stirred solution of
4-(dimethylaminomethyl)-9-benzyl-6-methox-
y-1,2,3,4-tetrahydrocarbazole (4 g, 0.01 mol) in liquid NH.sub.3
(300 mL). NH.sub.4Cl (3.0 g) was added until the blue-collar of the
mixture dissipated. The NH.sub.3 was evaporated, water (50 mL) was
added in the mixture was extracted with CH.sub.2CL.sub.3 The
combined organic portion was washed with water (50 mL), brine (50
mL), dried (MgSO.sub.4) and evaporated to give an oil. The oil was
purified by column chromatography (CHCL.sub.3/MeOH; 9:1) and
converted to an oxalate salt. The salt was recrystallized from
anhydrous Et.sub.2O/absolute EtOH to give 1.8 g of the desired
target as a white powder..sup.1H-NMR (CDCl.sub.3, free base) d 8.10
(s, 1H, NH), 7.20 (t, 1H, ArH), 6.90 (d, 1H, ArH), 6,70 (dd, 1H,
ArH), 3.80 (s, 3H, OCH3), 3.40 (t, 1H, CH), 3.15 (d, 1H, CH), 3.0
(t, 1H, CH)3.00 (t, 1H, Ch), 2.82 (s, 6H, 2.times.CH3), 2.63-2.73
(m, 2H, CH2), 2.23 (m, 1H, CH) 1.8-2.0 (m, 3H, CH2-CH).
EXAMPLE 7
[0040] A mixture of Compound G as free base (0.5 g, 1.94 mmol) and
sodium hydride (60%) (0.085 g, 3.54 mmol) was heated at 100.degree.
C. under nitrogen until the evolution of H.sub.2 gas ceased. The
resultant was dissolved in anhydrous DMF and
benzenesulfonylchloride (0.30 mL, 2.35 mmol) was added drop wise at
0.degree. C. The reaction mixture was stirred at room temperature
overnight. Saturated NaHCO.sub.3 solution was added and extracted
with CH.sub.2Cl.sub.2 (3.times.25 mL). The organic layer was dried
over MgSO.sub.4 and the solvent was removed under pressure. The
residue was purified by column chromatography
(CH.sub.2Cl.sub.2/MeOH; 9:1) as eluent to give an oil (0.60 g, 76%)
and converted to hydrochloride salt. The hydrochloride salt was
recrystallized from ethanol and anhydrous ether mp 259-261.degree.
C.
EXAMPLE 8
6,7,8,9-tetrahydro-2-methoxy-10-(N,N-dimethylaminoethyl)pyridol[1,2-a]indo-
le-9-one Oxalate (I)
[0041] A mixture of 5-methoxy-N,N-dimethyltryptamine (free base)
(2.00 g, 9.17 mmol) and 60% NaH (0.41 g, 10.1 mmol) was heated at
100.degree. C. under N.sub.2 until evolution of gas had ceased. The
resultant mass was dissolved in anhydrous DMF (25 ml) and anhydrous
g-butryolactone 1.4 mL, 18.2 mmol) was added in dropwise manner at
room temperature. The reaction mix was heated at reflux for 20 h,
cooled to 0.degree. C., and acidified by the addition of an
ethereal solution of HCl. Additional Et.sub.2O was added to the
resulting suspension and the supernatant was decanted. The residue
was dissolved in PPE (52.5 mL) and CHCl.sub.3 (100 mL) and the
reaction mixture was heated at reflux for 3 h under N.sub.2. The
resulting mixture was neutralized by the addition of 15% NaOH
solution at ice-bath temperature, and extracted with
CH.sub.2Cl.sub.2. The organic portion was dried (MgSO.sub.4) and
solvent was removed under reduced pressure. The residue was
purified by flash chromatography on silica gel
(CH.sub.2Cl.sub.2/MeOH; 20:1) to give 0.52 g (20%) of
6,7,8,9-tetrahydro-2-methoxy-10-(N,N-dimethylaminoethyl)pyridol[1,2-a]ind-
ole-9-One free base as a yellow oil. .sup.1H-NMR (DMSO-d.sub.6)
.quadrature.7.35 (d, 1H, J=8.79 Hz) 7.18 (s, 1H), 6.88 (d, 1H,
J=8.85 Hz), 4.06 (t, 2H, J=6.60 Hz), 3.80 (s, 3H), 3.42-3.36 (m,
2H), 3.17-3.12 (m, 2H), 2.85 (s, 6H), 2.66-2.62 (m, 2H0. IR
(CHCl.sub.3) 1648.sup.cm-1. A small sample was converted to the
oxalate salt; mp 191-192.degree. C.
EXAMPLE 9
6,7,8,9-tetrahydro-2-methoxy-10-(N,N-dimethylaminoethyl)pyridol[1,2-a]indo-
le Oxalate (J)
[0042] A solution of 1.0 M borane/THF (2 mL, 2 mmol) was added ion
a dropwise manner to ice-bath cooled
6,7,8,9-tetrahydro-2-methoxy-10-(N,N-d-
imethylaminoethyl)pyridol[1,2-a]indole-9-one Oxalate (290 mg, 1.01
mmol) under N.sub.2. The reaction mixture was allowed to stir at
room temperature for 2 h. Acetone (3 ml) was added , and the
reaction mixture was heated at reflux for 1 h to quench the
unreacted borane reagent. The solvent was removed under reduced
pressure. The residue was purified by flash chromatography on
silica gel (hexane/EtOAc; 4:1) to give 207 mg (75%) of a light
yellow oil. .sup.1H-NMR (DMSO-d.sub.6) .quadrature.7.34 (d, 1H,
J=8.85 Hz) 7.21 (s, 1H), 4.08 (t, 2H, J=6.65 Hz), 3.79 (s, 3H),
3.40-3.35 (m, 2H), 3.30-3.25 (m, 2H), 3.06-3.01 (m, 2H) 2.83 (s,
6H) 1.76-1.69 (m, 2H), 1.40-1.31 (m, 2H). A small portion was
converted to its oxalate salt; mp 114-115.degree. C.
EXAMPLE 10
1-Benzenesulfonyl-5-methoxy-N,N,-dimethyltryptamine Oxalate
[0043] A mixture of 5-methoxy-N,N-dimethyltryptamine (free base)
(4.35 g, 19.93 mmol) and 60% NaH (0.87 g, 21.75 mmol) was heated at
100.degree. C. under N.sub.2 until evolution of H.sub.2 gas ceased.
The resultant mass was dissolved in anhydrous DMF (21 ml) and
benzenesulfonyl chloride (2.8 ml, 21.94 mmol) and 60% NaH (0.87 g,
21.75 mmol) was added in a dropwise manner at 0.degree. C. The
reaction mixture was allowed to stir at room temperature overnight.
Saturated NaHCO.sub.3 solution was added and the mixture was
extracted with CH.sub.2Cl.sub.2. The organic portion was dried
(MgSO.sub.4)and the solvent was removed under reduced pressure. The
residue was purified by flash chromatography on silica gel
(CH.sub.2Cl.sub.2/MeOH; 9:1) to give 4.39 g of an oil (61%).
.sup.1H-NMR (CDCl.sub.3) .quadrature.7.89-7.87(m, 1H,) 7.83 (d, 2H,
J=8.0 Hz), 7.51 (t, 1H, J=7.8 Hz), 7.34 (s, 1H), 6.93-6.92 (m, 2H),
3.82 (s, 3H), 2.80 (t, 2H, J=7.8 Hz) 2.59 (t, 2H, J=7.8 Hz), 2.33
(s, 6H). IR CHCl.sub.3, 1357, 1115 c.sup.m-1. The oxalate salt was
prepared and recrystallized from an acetone/Et2O mixture; mp
224-226.degree. C.
EXAMPLE 11
1-Benzenesulfonyl-5-methoxy-2-n-propyl-N,N-dimethyl Tryptamine
[0044] A 2.5 M solution of nBuLi (1.4mL, 3.5 mmol) was added in a
dropwise manner to a stirred solution of
1-Benzenesulfonyl-5-methoxy-N,N,-dimethyl- tryptamine oxalate (free
base) (1.00 g, 2.79 mmol) in DME (4 mL) at -10.degree. C. under
N.sub.2 The resulting solution was allowed to stir for an
additional 10 min at -10.degree. C., and then nPrl (0.35 mL,
3.59mmll) was added. The reaction mixture was allowed to stir for 1
h at -10.degree. C. Saturated NaHCO.sub.3 solution was added and
the reaction mixture was extracted with CHCl2. The organic portion
was washed with brine and dried (MgSO.sub.4); the solvent was
removed under reduced pressure and the residue was purified by
flash chromatography on silica gel. (CH.sub.2Cl.sub.2/MeOH; 30:1).
To give 0.19 g of a bright yellow oil. .sup.1H-NMR (CDCl.sub.3)
.quadrature.8.06 (d, 1H, J=8.79 Hz), 7.62 (d, 2H, J=8.22 Hz),
7.51-7.46 (m, 1H), 6.89-6.85 (m, 1H), 3.85 (s, 3H), 2.96-2.89 (m,
4H) 2.63-2.57 (m, 2H), 2.48 (s, 6H) 6.87, 1.73 (q, 2H, J=7.51 Hz),
1.00 (t, 3H, J=7.51Hz). IR CHCl.sub.3, 1355 c.sup.m-1.
EXAMPLE 12
5-HT Derivative Binding to 5-HT.sub.6 Receptor
[0045] The binding assay employed human 5-HT.sub.6 stably
transfected to HEK 293 human embryonic kidney cells with [.sup.3H]
lysergic acid diethylamide (70 Ci/mmol; Dupont NEN) as radioligand.
Radioligand was diluted in incubation buffer in borosilicate glass
vials and protected from light. Competing agents (1 mM stock
solution) were dissolved in DMSO or saline and stored at -20 0 C.
Dilutions of compounds were made using incubation buffer in 96-well
plates and mixed by multichannel pipetting. Serial dilutions (1 in
4) started at a final concentration of 10,000 nM. Final
concentrations >10,000 nM were individually prepared from the 1
mM stock solution. Nonspecific binding was defined by 100 mM
serotonin creatinine sulfate (Research Biochemicals) prepared fresh
in incubation buffer at the time of each determination, and
protected from light. in incubation buffer at the time of each
determination, and protected from light. Reaction volumes were as
follows: 200 ml incubation buffer (50 mM tris, 0.5 mM EDTA, 10 mM
MgCl.sub.2), pH 7.4 at 22 0 C, 100 ml test agent or serotonin (100
mM) or buffer, 100 ml [3H]lysergic acid diethylamide (2 nM final
concentration) and 100 ml membrane preparation (15 mg protein). The
incubation was initiated by the addition of membrane homogenate and
the plates vortexed. The plates were incubated, with protection
from light, by shaking at 37 0 C for 60 min. The binding reaction
was stopped by filtration. The samples were filtered under vacuum
over 96 well glass fiber filters, presoaked in 0.3% PEI in 50 mM
tris buffer (pH 7.4) for at least 1H and then washed 6.times. with
1 ml of cold 50 mM tris (4 0 C, pH 7.4) using a Packard Filtermate
Harvester. The unifilter plates were dried overnight in a
37.degree. C. dry incubator. The unifilter bottoms were sealed and
35 ml of Packard MicroScint was added. The plates were allowed to
equilibrate for 1 h and were then sealed using a Packard TopSeal P
with the Packard Plate Micromate 496. Plates were counted by liquid
scintillation spectrometry. Each well was counted for 3 min.
Compounds were initially assayed at 1000 and 100 nM. If a compound
caused at least 80% inhibition of [.sup.3H]lysergic acid
diethylamide binding at 1000 nM, it was further tested and a Ki
determined. The range of concentrations chosen was so that the
middle concentration would produce approximately 50%
inhibition.
[0046] Table 1 shows the affinity for 5-HT.sub.6 receptors of
claimed compounds derived from the general formula in formula 2
above.
1 COMPOUND R1 R2 R3 R4 R5 Ki(nm) A Me Me 5-OMe H Me 60 B Me Me
5-OMe H Et 16 C Me Me 5-OMe H nPr 185 D Me Me 5-OMe H Phenyl 20
[0047] Table 2 shows the results of affinity for 5-HT.sub.6
receptors of claimed compounds derived from the general formula in
formula 3 above.
2 COMPOUND R1 and R2 R4 Ki(nm) E Me --CH2-phenyl 136 F H
--Ch2-phenyl 302 G Me H 168 H Me --SO2-phenyl 2
[0048] Table 3 shows the results of affinity for 5-HT.sub.6
receptors of claimed compounds derived from the general formula in
formula 4 above.
3 COMPOUND X Ki(nM) I O 84 J H2 1030
EXAMPLE 13
Characterization of 5HT.sub.6 Selectivity
[0049] Selected compounds were examined to determine their
specificity of binding to the 5-HT.sub.6 receptor. Compounds were
examined at more than 30 receptor populations. Assays for the
following receptors were performed as per the NIMH Psychoactive
Drug Screening Program. The compounds failed to displace
radioligand (i.e., <50% displacement) at a concentration of
10,000 nM at most receptors. Where more than 50% displacement was
observed, Ki values were determined and the data are reported in
the following table. It can be seen that the compounds are
selective for 5-HT.sub.6 receptors.
[0050] Table 4 shows the 5-selectivity of several compounds of the
invention.
4TABLE 4 Ki, nM(SEM) Receptor COMPOUND COMPOUND COMPOUND Population
A B D CONTROL AND AGENT NET 6,380(3190) >10,000 >10,000
Nortriptyline 6.3(1.2) SERT >10,000 >10,000 4,700(1550)
Fluoxetine 3.5(0.7) h5-HT1A 200(60) 170(54) 1,470(310) WAY 1000,635
0.6(1.5) h5-HT1D 250(180) 290(700) 6,225(70) Ergotamine 0.8(0.6)
h5-HT1E 1,800(600) 520(180) >10,000 Serotonin 0.5(.015) r5-HT2A
>10,000 >10,000 470(10) Clozapine 9(1) r5-HT2C 4,020(640)
1,810(490) 675(180) Clozapine 23(5) h5-HT5A 10,450(2195) 4,620(650)
5,160(930) Ergotamine 22(3) h5-HT7 145(34) 300(60) 155(35)
Clozapine 9(2) h5-Ht6 60(13) 16(4) 20(5) Clozapine 10(3)
[0051] Compounds displayed Ki values of >10,000 nM at the
following populations of receptors: histamine, NMDA, PCP,
acetylcholine, opiate, and vasopressin receptors. Ki values were
>10,000 nM for compounds A and B at hD1, rD2, rD3, rD4, and hD5
receptors and 10,000 nM for D at hD1, rD2, and rD4 receptors.
Compound D produced 70% inhibition at rD3 and hD5 receptors. NET
and SERT represent the norepinephrine and serotonin transporters.
Ki values for all three compounds were >10,000 at the dopamine
transporter.
EXAMPLE 14
cAMP Activation Assays
[0052] Human 5-HT.sub.6 receptors stably expressed in 293 HEK cells
were grown in 24-well plates to near confluence and 18 h prior to
the assay the medium was replaced with DMEM containing dialyzed 10%
Fetal Calf Serum. For the assay, the medium was aspirated and
replaced with fresh DMEM without serum and incubated with various
concentrations of compounds of the invention in a total volume of
0.5 ml for 15 min. The assay was terminated by aspiration and the
addition of 10% trichloroacetic acid (TCA). The TCA extract was
used for cAMP determinations. (Data represent the mean of N=4
separate determinations). Results of cAMP activation by various
compounds of the invention are shown in the attached Drawing of
FIG. 1.
[0053] While the invention has been described in terms of its
preferred embodiments, those skilled in the art will recognize that
the invention can be practiced with modification within the spirit
and scope of the appended claims. Accordingly, the present
invention should not be limited to the embodiments as described
above, but should further include all modifications and equivalents
thereof within the spirit and scope of the description provided
herein.
* * * * *