U.S. patent application number 14/276195 was filed with the patent office on 2014-08-28 for synthesis of functionalized octahydro-isoquinolin-1-one-8-carboxylic esters and analogs, and therapeutic methods.
This patent application is currently assigned to University of Kansas. The applicant listed for this patent is University of Kansas, University of North Carolina. Invention is credited to Jeffrey Aube, Kevin J. Frankowski, Partha Ghosh, Bryan L. Roth.
Application Number | 20140243333 14/276195 |
Document ID | / |
Family ID | 42060433 |
Filed Date | 2014-08-28 |
United States Patent
Application |
20140243333 |
Kind Code |
A1 |
Aube; Jeffrey ; et
al. |
August 28, 2014 |
SYNTHESIS OF FUNCTIONALIZED
OCTAHYDRO-ISOQUINOLIN-1-ONE-8-CARBOXYLIC ESTERS AND ANALOGS, AND
THERAPEUTIC METHODS
Abstract
A functionalized polycyclic compound can have a structure of
Formula 1 or salt, prodrug, analog, or derivative thereof, which
compound can be prepared by providing a diene; reacting the diene
with a dienophile under sufficient conditions for a combined
Diels-Alder/acylation reaction so as to provide a polycyclic
compound having a carboxylic acid; and coupling the carboxylic acid
with an amine-containing compound or a hydroxyl-containing compound
so as to form an amide or an ester and producing a compound having
a structure of Formula 1. The compound can be used for modulating
an opioid receptor, which can be conducted by administering to an
opioid receptor a functionalized polycyclic compound as described
herein in an effective amount to modulate the functionality of the
opioid receptor. Such opioid modulation can provide a biological
benefit to a subject.
Inventors: |
Aube; Jeffrey; (Lawrence,
KS) ; Roth; Bryan L.; (Durham, NC) ; Ghosh;
Partha; (Lawrence, KS) ; Frankowski; Kevin J.;
(Lawrence, KS) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
University of Kansas
University of North Carolina |
Lawrence
Chapel Hill |
KS
NC |
US
US |
|
|
Assignee: |
University of Kansas
Lawrence
KS
University of North Carolina
Chapel Hill
NC
|
Family ID: |
42060433 |
Appl. No.: |
14/276195 |
Filed: |
May 13, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12567573 |
Sep 25, 2009 |
8735391 |
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14276195 |
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61100619 |
Sep 26, 2008 |
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Current U.S.
Class: |
514/235.2 ;
514/253.05; 514/309; 544/128; 544/363; 546/141 |
Current CPC
Class: |
A61P 25/06 20180101;
A61P 25/30 20180101; C07D 413/06 20130101; A61P 25/24 20180101;
A61P 13/00 20180101; A61P 25/20 20180101; A61P 9/00 20180101; C07D
401/06 20130101; C07D 417/12 20130101; A61K 31/472 20130101; A61P
13/12 20180101; A61P 25/16 20180101; A61P 29/00 20180101; C07D
217/22 20130101; C07D 217/24 20130101 |
Class at
Publication: |
514/235.2 ;
546/141; 544/128; 544/363; 514/309; 514/253.05 |
International
Class: |
C07D 217/24 20060101
C07D217/24; C07D 401/06 20060101 C07D401/06; C07D 417/12 20060101
C07D417/12; C07D 413/06 20060101 C07D413/06 |
Claims
1. A functionalized polycyclic compound comprising: a structure of
Formula 1 or salt, prodrug, analog, enantiomer, or derivative
thereof, wherein: ##STR00128## R1 and R3 are independently nothing,
hydrogen, halogen, hydroxyl, straight or branched substituted or
unsubstituted alkoxy, amine, straight or branched substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted aryl, branched or unbranched or cyclic
substituted or unsubstituted arylalkyl, or combinations thereof;
R2, R4, R5, R6, R11, R12, R13, R14, R15, R16, R17, and R18 are
independently a hydrogen, halogen, hydroxyl, straight or branched
substituted or unsubstituted alkoxy, amine, straight or branched
substituted or unsubstituted alkyl, substituted or unsubstituted
cycloalkyl, substituted or unsubstituted aryl, branched or
unbranched or cyclic substituted or unsubstituted arylalkyl, or
combinations thereof, or one or more of R4 and R5 together, R11 and
R12 together, R13 and R14 together, and R15 and R16 together form a
bond or a ring therebetween; R7 and R8 are both a hydrogen or
together form a bond therebetween, or R7 and R8 together are O; R9
and R10 are each independently an O or two separate hydrogen atoms;
X1 and X2 independently are O, N, or S; n is from 0 to 5; when X1
is N and R1 is nothing, then R2 is a ring with the N; when X1 is O
and R1 is nothing, then R2 is as defined; and when X2 is N, R3 is a
something.
2. A compound as in claim 1, when any of R1-R3 includes an aryl,
the aryl is substituted with one or more electron withdrawing
groups.
3. A compound as in claim 2, wherein the electron withdrawing
groups are selected from Br, Cl, I, and CF3.
4. A compound as in claim 1, wherein the compound has a structure
of one of Formulas 2-16. ##STR00129## ##STR00130## ##STR00131##
5. A compound as in claim 1, wherein X1 and X2 are both N.
6. A compound as in claim 1, wherein X1 is N and X2 is O.
7. A compound as in claim 1, wherein X1 is O and X2 is N.
8. A compound as in claim 1, wherein X1 and X1 are both O.
9. A compound as in claim 1, wherein one or more of R1, R2, R3, R4,
R5, R6, R11, R12, R13, R14, R15, R16, R17, and R18 is one of the
following side chains (side chains 1-16): ##STR00132##
##STR00133##
10. A compound as in claim 1, where the compound is selected from
the Compounds of Tables 4, 5, 8, and 9.
11. A method for preparing a functionalized polycyclic compound,
the method comprising: providing a hydroxyl-containing diene or an
amine-containing diene; reacting the diene with a dienophile under
sufficient conditions for a combined Diels-Alder/acylation reaction
so as to provide a polycyclic compound having a carboxylic acid;
and conjugating the carboxylic acid with an amine-containing
compound or a hydroxyl-containing compound so as to form an amide
or an ester and producing a compound having a structure of Formula
1 or derivative thereof, wherein: ##STR00134## R1 and R3 are
independently nothing, hydrogen, halogen, hydroxyl, straight or
branched substituted or unsubstituted alkoxy, amine, straight or
branched substituted or unsubstituted alkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted aryl,
branched or unbranched or cyclic substituted or unsubstituted
arylalkyl, or combinations thereof; R2, R4, R5, R6, R11, R12, R13,
R14, R15, R16, R17, and R18 are independently a hydrogen, halogen,
hydroxyl, straight or branched substituted or unsubstituted alkoxy,
amine, straight or branched substituted or unsubstituted alkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted aryl, branched or unbranched or cyclic substituted or
unsubstituted arylalkyl, or combinations thereof, or one or more of
R4 and R5 together, R11 and R12 together, R13 and R14 together, and
R15 and R16 together form a bond or a ring therebetween; R7 and R8
are both a hydrogen or together form a bond therebetween, or R7 and
R8 together are O; R9 and R10 are each independently an O or two
separate hydrogen atoms; X1 and X2 independently are O, N, or S; n
is from 0 to 5; when X1 is N and R1 is nothing, then R2 is a ring
with the N; when X1 is O and R1 is nothing, then R2 is as defined;
and when X2 is N, R3 is a something.
12. A method as in claim 11, wherein the Diels-Alder/acylation is
conducted at a temperature of at least about 25 degrees C. for a
duration of at least about 5 minutes.
13. A method as in claim 11, wherein the combined
Diels-Alder/acylation reaction is performed in an organic
solvent.
14. A method as in claim 13, wherein the solvent is
dichloroethane.
15. A method as in claim 11, wherein the combined
Diels-Alder/acylation reaction is at least near-neat.
16. A method as in claim 11, wherein the conjugation of the
carboxylic acid is catalyzed with a catalyst.
17. A method as in claim 16, wherein the catalyst is DMAP and/or
EDC-HCl.
18. A method as in claim 11, wherein the diene is an
amine-containing diene.
19. A method as in claim 11, wherein the diene is a
hydroxyl-containing diene.
20. A method as in claim 11, wherein the dienophile is a maleic
anhydride or citraconic anhydride.
21. A method for preparing a functionalized polycyclic compound
having a carboxylic acid, the method comprising: providing a
hydroxyl-containing diene or an amine-containing diene; and
reacting the diene with a dienophile under sufficient conditions
for a combined Diels-Alder/acylation reaction so as to provide a
polycyclic compound having a carboxylic acid having Formula 17:
##STR00135## R3, R4, R5, and R6 are independently a hydrogen,
halogen, hydroxyl, straight or branched substituted or
unsubstituted alkoxy, amine, straight or branched substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted aryl, branched or unbranched or cyclic
substituted or unsubstituted arylalkyl, or combinations thereof, or
R4 and R5 together form a ring therebetween; X is an O, N, or S;
and n is from 0 to 5.
22. A method as in claim 22, further comprising preparing the
hydroxyl-containing diene or the amine-containing diene.
23. A method as in claim 22, therein the aminodiene component is
prepared from 3,5-hexadien-1-ol.
24. A pharmaceutical composition comprising: a pharmaceutically
acceptable carrier; and the functionalized polycyclic compound in
accordance with claim 1.
25. A method of modulating an opioid receptor, the method
comprising: administering to an opioid receptor the functionalized
polycyclic compound in accordance with claim 1 in an effective
amount to modulate the functionality of the opioid receptor.
26. A method as in claim 25, wherein the opioid receptor is
selected from a delta, mu, kappa, or nociceptin opioid
receptor.
27. A method as in claim 25, wherein the functionalized polycyclic
compound agonizes the opioid receptor.
28. A method as in claim 25, wherein the functionalized polycyclic
compound antagonizes the opioid receptor.
29. A method of providing a therapy to a subject, the method
comprising: administering to the subject having an opioid receptor
the functionalized polycyclic compound in accordance with claim 1
in a therapeutically effective amount to modulate the functionality
of the opioid receptor so as to provide a biological benefit to the
subject.
30. A method as in claim 29, wherein the biological benefit is one
or more of the following: treatment for alcoholism; treatment for
drug addiction; reversing side effects or overdose of a MOR agonist
or opioid; treatment for obesity; treatment for Parkinson-induced
tardive dyskinesia; providing analgesic treatment; providing
antidepressant treatment; providing an anorectic treatment;
providing weight loss; treatment for a mood disorder; treatment for
bipolar disorders; treatment for drug seeking behavior; treatment
for stress-induced drug seeking behavior; providing spinal
analgesia; providing sedation; providing miosis; inhibiting ADH
release; providing respiratory depression; providing euphoria;
providing reduced GI motility; treatment of heart failure;
treatment of migraines; treatment of a variety of inflammatory
disorders; treatment of renal disorders; treatment of
cardiovascular disorders; treatment of psychotic disorders and
combinations thereof.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This U.S. patent application claims benefit of U.S.
provisional patent application having Ser. No. 61/100,619, filed on
Sep. 26, 2008, which provisional application is incorporated herein
by specific reference in its entirety.
BACKGROUND OF THE INVENTION
[0002] Opioid receptors are a group of G-protein coupled receptors
that have been identified to have opioids as ligands, which are
about 70% identical with differences located at N and C termini of
the receptors. Accordingly, opioid drugs have been developed to
exploit the biological activity of activating these receptors.
Examples of the opioid receptor types, subtypes, location, and
agonist activation activities are shown in Table A below. It has
been found that modulating the activity of the opioid receptors
through an agonist or antagonist can have different therapeutic
benefits. Various types of substances, ranging from small molecules
through polypeptides, have been explored to identify new drugs that
behave as agonists or antagonists of the opioid receptors.
TABLE-US-00001 TABLE A Receptor Location Function delta (.delta.)
Brain analgesia OP.sub.1.sup.(I) pontine nuclei antidepressant
effects amygdala olfactory bulbs deep cortex kappa (.kappa.) Brain
Spinal analgesia OP.sub.2.sup.(I) hypothalamus sedation
periaqueductal gray miosis claustrum inhibition of ADH Cortex
release Hippocampus spinal cord substantia gelatinosa mu (.mu.)
Brain Supraspinal analgesia OP.sub.3.sup.(I) cortex (laminae III
and IV) respiratory depression thalamus miosis striosomes euphoria
periaqueductal gray reduced GI motility spinal cord substantia
gelatinosa Nociceptin Brain treat heart failure receptor cortex
treat migraines OP.sub.4 amygdala appetite hippocampus development
of septal nuclei tolerance to .mu. agonists habenula hypothalamus
spinal cord
[0003] In addition to the activation of opioid receptors,
inactivation or antagonism of opioid receptors by antagonists can
also provide beneficial therapeutic effects. For example, a delta
opioid receptor (DOR) antagonist may be useful for treatment for
alcoholism and depression. A kappa opioid receptor (KOR) antagonist
may be useful for a treatment for drug addiction, depression,
inflammation, gastrointestinal, and renal diseases. A mu opioid
receptor (MOR) antagonist may be useful for reversing side effects
or overdose of a MOR agonist or opioid, obesity, and 1-DOPA induced
dyskinesia in Parkinson's Disease. A nociceptin receptor (NR)
antagonist may be useful for providing analgesic and antidepressant
biological activity.
[0004] Other therapeutic benefits and biological activities of
opioid receptors continue to be studied. Accordingly, it is
important to develop new compounds that have activity with opioid
receptors. While traditional chemistry techniques can be applied to
creating new compounds, it is also desirable to improve chemical
synthesis techniques. This can include simplifying the synthetic
chemistry so that compounds can be prepared more easily.
[0005] Therefore, it would be advantageous to develop new compounds
that were active with opioid receptors. Additionally, it would be
beneficial to have improved synthesis techniques so that new
compounds can be prepared easier and with more straight forward
synthesis protocols.
BRIEF SUMMARY OF THE INVENTION
[0006] In one embodiment, the present invention includes
functionalized polycyclic compounds having a structure of Formula 1
or salt, prodrug, analog, or derivative thereof, wherein Formula 1
is shown below. The structure of Formula 1 can be defined as
follows: R1 and R3 are independently nothing, hydrogen, halogen,
hydroxyl, straight or branched substituted or unsubstituted alkoxy,
amine, straight or branched substituted or unsubstituted alkyl,
substituted or unsubstituted cycloalkyl, substituted or
unsubstituted aryl, branched or unbranched or cyclic substituted or
unsubstituted arylalkyl, or combinations thereof; R2, R4, R5, and
R6 are independently a hydrogen, halogen, hydroxyl, straight or
branched substituted or unsubstituted alkoxy, amine, straight or
branched substituted or unsubstituted alkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted aryl,
branched or unbranched or cyclic substituted or unsubstituted
arylalkyl, or combinations thereof, or R4 and R5 together form a
bond or a ring therebetween; R7 and R8 are both a hydrogen or
together form a bond therebetween, or R7 and R8 together can be O
(e.g., an epoxide); R9 and R10 are each independently an O or two
separate hydrogen atoms; X1 and X2 independently are O, N, or S; n
is from 0 to 5; when X1 is N and R1 is nothing, then R2 is a ring
with the N; when X1 is O and R1 is nothing, then R2 is as defined;
and when X2 is N, R3 is a something. Also, the compound can be
defined as follows: wherein X1 and X2 are both N; X1 is N and X2 is
O; X1 is O and X2 is and X1 and X1 are both O. In one embodiment,
R1-R3 can include an aryl; the aryl is substituted with one or more
electron withdrawing groups. For example, the electron withdrawing
groups are selected from Br, Cl, I, and CF.sub.3.
[0007] In one embodiment, the compound has a structure of one of
Formulas 2-16 shown below.
[0008] In one embodiment, one or more of R1, R2, R3, R4, R5, or R6
is one, of chains 1-16 shown below.
[0009] In one embodiment, the compound is selected from the
compounds of Tables 4, 5, 8, and 9.
[0010] In one embodiment, the present invention includes a method
for preparing a functionalized polycyclic compound. Such a method
can include: providing a diene; reacting the diene with a
dienophile under sufficient conditions for a combined
Diels-Alder/acylation reaction so as to provide a polycyclic
compound having a carboxylic acid; and coupling the carboxylic acid
with an amine-containing compound or a hydroxyl-containing compound
so as to form an amide or an ester and producing a compound having
a structure of Formula 1 (as shown and defined herein) or
derivative thereof.
[0011] In one embodiment, the Diels-Alder/acylation is conducted at
a temperature of at least about 165 degrees C. (+/-10-20 degrees
C.) for a duration of at least about 1.5 hours (+/-15-30 minutes)
when using a solvent. For neat or near neat conditions, no external
heating is required, and the reaction is exothermic to provide
heat, and thereby, the heat and time of a neat or near neat
reaction protocol can vary, and some results have produced
sufficient reactions in as little as 10 minutes.
[0012] In one embodiment, the combined Diels-Alder/acylation
reaction is performed in an organic solvent. For example, the
solvent can be dichloroethane or toluene. Optionally, the combined
Diels-Alder/acylation reaction is at least near-neat. Also, the
reaction can be performed without solvent. The reaction can be
exothermic so as to provide an increase in temperature.
[0013] In one embodiment, the coupling of the carboxylic acid can
be catalyzed with a catalyst. For example, the catalyst can be DMAP
and/or a coupling reagent such as EDC-HCl. The diene of the
reaction can be an amine-containing diene or a hydroxyl-containing
diene. The dienophile can be a maleic anhydride or citraconic
anhydride or the like.
[0014] In one embodiment, the invention can include a method for
preparing a functionalized polycyclic compound having a carboxylic
acid. Such a method can include providing a hydroxyl-containing
diene or an amine-containing diene; and reacting the diene with a
dienophile under sufficient conditions for a combined
Diels-Alder/acylation reaction so as to provide a polycyclic
compound having a carboxylic acid having Formula 17 as shown below.
In Formula 17, R3, R4, R5, and R6 can be independently a hydrogen,
halogen, hydroxyl, straight or branched substituted or
unsubstituted alkoxy, amine, straight or branched substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted aryl, branched or unbranched or cyclic
substituted or unsubstituted arylalkyl, or combinations thereof, or
R4 and R5 together form a ring therebetween; X can be an O or N;
and n can be from 0 to 5. The method can also include preparing the
hydroxyl-containing diene or the amine-containing diene. For
example, the aminodiene component can be prepared from
3,5-hexadien-1-ol or the like.
[0015] In one embodiment, the invention can include a
pharmaceutical composition having a pharmaceutically acceptable
carrier, and a functionalized polycyclic compound.
[0016] In one embodiment, the invention can include a method of
modulating an opioid receptor, which can be conducted by
administering to an opioid receptor a functionalized polycyclic
compound as described herein in an effective amount to modulate the
functionality of the opioid receptor. The modulated opioid receptor
can be selected from a delta, mu, kappa, or nociceptin opioid
receptor. The compound can be an opioid agonist or antagonist.
[0017] In one embodiment, the invention can include a method of
providing a therapy to a subject, which can be conducted by
administering to the subject having an opioid receptor a
functionalized polycyclic compound as described herein in a
therapeutically effective amount to modulate the functionality of
the opioid receptor so as to provide a biological benefit to the
subject. For example, the biological benefit is one or more of the
following: treatment for alcoholism; treatment for drug addiction;
reversing side effects or overdose of a MOR agonist or opioid;
treatment for obesity; treatment for L-DOPA-induced dyskinesia in
Parkinson's Disease; providing analgesic treatment; providing
antidepressant treatment; providing an anorectant treatment;
providing weight loss; treatment for a mood disorder; treatment for
bipolar disorders; treatment for psychotic disorders; treatment for
drug seeking behavior; treatment for stress-induced drug seeking
behavior; providing spinal analgesia; providing sedation; providing
miosis; inhibiting ADH release; providing respiratory depression;
providing euphoria; providing reduced GI motility; treatment of
heart, failure; treatment of migraines; treatment of a variety of
inflammatory disorders; treatment of renal disorders; treatment of
cardiovascular disorders; and combinations thereof.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0018] Generally, the present invention relates to new compounds of
general Formulas 1-16 below. The compounds generically are
functionalized polycyclic analogs with amide or ester functional
groups, and can be included in analog libraries. The compounds can
include a two-ring polycycle, wherein one homocyclic ring has 6
carbon atoms fused to a heterocyclic ring of 5-8 atoms with the
hetero atoms being N or O, and where the heterocyclic ring can be
saturated or include a ketone. The homocycle and heterocycle can
both include functional groups. The homocycles can be cycloalkyls,
cycloalkenes, cyclohexanes, cyclohexenes and the like. The
heterocycles can be piperidines, 2-piperidones, lactones,
tetrahydropyrans, and corresponding heterocycles having more or
less atoms in the cycle. The compounds can be referred to as
N-alkyl-octahydroisoquinolin-1-one-8-carboxyls and analogs
thereof.
[0019] Also, the present invention includes new methods of
synthesizing the compounds, such as a modified
Diels-Alder/acylation reaction scheme and library production
techniques. The compounds can be bioactive with regard to
interacting and modulating opioid receptors, and may be agonists
and/or antagonists. The new compounds can be biologically active
with opioid receptors, such as being agonists with high activity or
low activity or as antagonists. Some compounds can be broad
spectrum agents that interact with more than one opioid receptor to
provide biological activity. Other compounds can have more
targeted, specific activities to a preferred opioid receptor, which
can be important when selectivity and avoidance of side effects is
desirable. Additionally, the compounds may be biologically active
with other enzymes, receptors, or other proteins. The agonists and
antagonists of opioid receptors can be used in therapeutic
protocols to provide a treatment to a patient.
[0020] Additionally, the new compounds can be prepared with simpler
synthetic protocols, which can make creating analog libraries a
more straight forward process. The simpler synthetic protocols can
include combining reaction steps so that the synthesis is easier,
such as by using domino reaction synthetic schemes.
[0021] Domino reactions, in which multiple chemical reactions are
carried out in a single step, are attractive tools for library
synthesis because they can lead to complex structures quickly and
with a minimum of chemical manipulations. The domino reactions can
be used to generate scaffolds that contain reactive sites that can
be used for subsequent modification and analog production, which
leads to focused chemical libraries. The use of a strategy-level
carbon-carbon bond forming reaction, such as the Diels-Alder
cycloaddition, is attractive for library synthesis because of its
well-known scope and ability to lead to cyclic materials containing
multiple stereocenters. Accordingly, new synthetic reaction
protocols have been developed that combine the Diels-Alder reaction
with an imide acylation reaction to produce functionalized
polycyclic-carboxylic acid analogs. The utility of this reaction
scheme has been demonstrated by the synthesis of a small solution
phase focused library of compounds, which are described herein. The
carboxylic acid can then be reacted with various functional groups
to provide a library. For example, a classical Diels-Alder reaction
can be modified so that a reaction of a maleic anhydride (or other
dienophile) with an amine-containing diene can be performed in a
single step, such as is shown in Scheme 1 below. In this reaction
protocol, the Alder-endo rule can result in the amine-containing
side chain emerging cis to a reactive carbonyl group. This allows
for both the Diels-Alder and the acylation steps to be performed
simultaneously in a tandem, one-pot synthesis of functionalized
polycyclic analogs that contain a carboxylic acid, which allows for
diversification in a second step. That is, the modified Diels-alder
reaction can produce a scaffold having a reactive carboxylic acid
group that can then be synthetically transformed to different
functional groups to obtain an analog library.
[0022] It was surprising and unexpected that the modified
Diels-Alder reaction could successfully produce functionalized
polycyclic-carboxylic acids with a reactive group for simple
conjugation chemistry. Previously, much work in this area has
focused on intramolecular versions in which the diene and
dienophiles are attached prior to cycloaddition, which is
unfavorable in comparison to the modified Diels-Alder reaction
described herein. Accordingly, it is surprising and unexpected that
the chemical synthesis described herein can include the use of an
extremely reactive dienophile in order to form the C--N bond
without a separate alkylation event, and the availability to
provide a carboxylic acid group for downstream manipulation.
[0023] Additionally, new reactions are described herein for
preparing functionalized polycyclic esters and lactones.
A. DEFINITIONS
[0024] As used herein, the terms "an effective amount",
"therapeutic effective amount", or "therapeutically effective
amount" shall mean an amount or concentration of a compound
according to the present invention which is effective within the
context of its administration or use. Thus, the term "effective
amount" is used throughout the specification to describe
concentrations or amounts of compounds according to the present
invention which may be used to produce a favorable change in the
disease or condition treated, inhibited, or prevented, whether that
change is a remission, a decrease in desire for a drug such as
cocaine or in addiction characteristics, a favorable physiological
result, or the like, depending upon the disease or condition
treated.
[0025] As used herein, the term "pharmaceutically acceptable
excipient" means an excipient that is useful in preparing a
pharmaceutical composition that is generally safe, non-toxic and
neither biologically nor otherwise undesirable, and includes an
excipient that is acceptable for veterinary use as well as human
pharmaceutical use. A "pharmaceutically acceptable excipient" as
used in the specification and claims includes both one and more
than one such excipient.
[0026] As used herein, the term "coadministration" or "combination
therapy" is used to describe a therapy in which at least two active
compounds in effective amounts are used for the treatment,
inhibition, and/or prevention of a condition.
[0027] As used herein, the term "treating" or "treatment" of a
disease, including drug addiction and drug-seeking behavior,
includes: (a) preventing the disease, i.e. causing the clinical
symptoms of the disease not to develop in a mammal that may be
exposed to or predisposed to the disease but does not yet
experience or display symptoms of the disease; (b) inhibiting the
disease, i.e., arresting or reducing the development of the disease
or its clinical symptoms; or (c) relieving the disease, i.e.,
causing regression of the disease or its clinical symptoms.
[0028] As used herein, the term "unit dosage form," refers to
physically discrete units suitable as unitary dosages for human and
animal subjects, each unit containing a predetermined quantity of
pharmacological agent calculated in an amount sufficient to produce
the desired effect in association with a pharmaceutically
acceptable diluent, carrier or vehicle.
[0029] As used herein, a "subject" or a "patient" refers to any
animal (preferably, a human), and preferably a mammal. Examples of
a subject or patient include a human, a non-human primate, a cow, a
horse, a pig, a sheep, a goat, a dog, a cat or a rodent such as a
mouse, a rat, a hamster, or a guinea pig. Generally, the invention
is directed toward use with humans.
Chemical Structures
[0030] Generally, the compounds of the present inventions are
functionalized polycyclic analogs and derivatives. Preferably, the
compounds modulate the activity of an opioid receptor, such as a
DOR, KOR, MOR, or NR. The compounds of the invention can generally
be described by the scaffolds in Formulas 1-16 shown below.
##STR00001## ##STR00002## ##STR00003##
[0031] In Formulas 1 through 16, R1 and R3 can be nothing,
hydrogen, halogen, hydroxyl, straight or branched substituted or
unsubstituted alkoxy, amine, straight or branched substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted aryl, branched or unbranched or cyclic
substituted or unsubstituted arylalkyl, or combinations thereof,
where the alkyls can be C1-C20, C1-C10, C1-C6, or C1-C4, where the
rings can be 3, 4, 5, 6, 7, or 8 membered, and any can include
chain atoms having hetero atoms selected from N, O, S, or P. R2,
R4, R5, R6, R11, R12, R13, R14, R15, R16, R17, and R18 can
independently be hydrogen, halogen, hydroxyl, straight or branched
substituted or unsubstituted alkoxy, amine, straight or branched
substituted or unsubstituted alkyl, substituted or unsubstituted
cycloalkyl, substituted or unsubstituted aryl, branched or
unbranched or cyclic substituted or unsubstituted arylalkyl, or
combinations thereof, where the alkyls can be C1-C20, C1-C10,
C1-C6, or C1-C4, where the rings can be 3, 4, 5, 6, 7, or 8
membered, and any can include chain atoms having hetero atoms
selected from N, O, S, or P. R4 and R5 together, R11 and R12
together, R13 and R14 together, and R15 and R16 together can form a
bond or a ring therebetween. R7 and R8 are both a hydrogen or
together form a bond therebetween, or R7 and R8 together can be O
(e.g., epoxide). R9 and R10 independently can be an O or two
separate hydrogen atoms. X1 and X2 independently can be O or N. The
"n" can be from 0 to 5, with 0, 1, and 2 preferred, with 1 most
preferred. When X1 is N and R1 is nothing, then R2 is a ring with
the nitrogen. When X1 is O and R1 is nothing, then R2 is as
defined. When X2 is N, R3 is something, such as described for R2.
The compounds can also be stereoisomers of the compounds
illustrated.
[0032] In one embodiment, R14 and R15 are hydrogen. It can also be
preferred that one or more of R11-R18 are hydrogen. However, any of
these R groups can be substituted as described herein.
[0033] In one embodiment, when any of R1-R3 includes an aryl, the
aryl can be substituted with one or more electron withdrawing
groups, such as Br, Cl, I, CF3 (halogenated alkyl) or the like,
especially for an R2 and/or R3.
[0034] In any of the compounds described herein, an R group alkyl
or aryl backbone carbon can be substituted with O, N, S, or P, with
O and N preferred. Any alkyl carbon can have a hydrogen replaced
with a halogen, Cl, F, CH.sub.3, CH.sub.3CH.sub.2, or higher or
lower substituted or unsubstituted straight chain or branched
aliphatic (e.g., C1-C10), an adamantyl (e.g., 2-adamantyl or
adamantane derivative), or cycle or heterocycle. R1-R6 can
independently include one or more fused or unfused cycles or
heterocycles selected from phenyl, pyridine, pyrimidine, pyrazine,
1,3,5-triazine, 1,2,4-triazine, quinoline, isoquinoline, acridine,
phenanthrolines, benzoquinolines, phenathridines, phenazines,
quinoxalines, quinazolines, phthalazines, pteridines, cinnolines,
pyrroles, imidazoles, 1,2,3-triazoles, 1,2,4, triazoles,
tetrazoles, isoxazoles, 1,3-thiazoles, benzimidazoles, indoles,
indazoles, benzothiazoles, phenols, naphthols, 2-furan, 3-furan,
2-thiophene, 3-thiophene, 2-pyrrole, 3-pyrrole, 2-oxazole,
4-oxazole, 5-oxazole, 2-thiazole, 4-thiazole, 5-thiazole,
2-imidazole, 4-imidazole, 5-imidazole, 3-isoxazole, 4-isoxazole,
5-isoxazole, 3-isothiazole, 4-isothiazole, 5-isothiazole, 4-(1,2,3)
oxadiazole, 5-(1,2,3) oxadiazole, 4-(1,2,3)triazole,
5-(1,2,3)triazole, or 2-(1,3,4) thiadiazole. Also, R1-R5 can
independently include amino acids or polypeptides.
[0035] Any of the compounds of Formulas 1-16 can include any one of
R1, R2, R3, R4, R5, or R6 being one of the following side chains
(side chains 1-16):
##STR00004## ##STR00005##
[0036] The compounds described herein can be prepared into racemic
mixtures, or the individual enantiomers thereof. Each compound can
also be present as an individual enantiomer that is separate from
its other enantiomers. It is thought that an individual enantiomer
may have enhanced biological activity over its other
enantiomers.
[0037] As used herein, the term "alkyl" or "aliphatic" can refer to
a hydrocarbyl moiety, such as an hydrocarbon group, that can be
straight or branched, saturated or unsaturated, and/or substituted
or unsubstituted, which has twenty or less carbons in the backbone.
An aliphatic group may comprise moieties that are linear, branched,
cyclic and/or heterocyclic, and contain functional groups such as
ethers, ketones, aldehydes, carboxylates, and the like. Exemplary
aliphatic groups include but are not limited to substituted and/or
unsubstituted groups of methyl, ethyl, propyl, butyl, pentyl,
hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl,
tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl,
nonadecyl, eicosyl, alkyl groups of higher number of carbons and
the like, as well as 2-methylpropyl, 2-methyl-4-ethylbutyl,
2,4-diethylpropyl, 3-propylbutyl, 2,8-dibutyldecyl,
6,6-dimethyloctyl, 6-propyl-6-butyloctyl, 2-methylbutyl,
2-methylpentyl, 3-methylpentyl, 2-ethylhexyl, and the like. The
terms aliphatic or alkyl also encompasses alkenyl groups, such as
vinyl, allyl, aralkyl and alkynyl groups.
[0038] Substitutions within an alkyl or aliphatic group can include
any atom or group that can be tolerated in the aliphatic moiety,
including but not limited to halogens, sulfurs, thiols, thioethers,
thioesters, amines (primary, secondary, or tertiary), amides,
ethers, esters, alcohols, oxygen, and the like. The aliphatic
groups can by way of example also include modifications such as azo
groups, keto groups, aldehyde groups, carbonyl groups, carboxyl
groups, nitro, nitroso or nitrile groups, heterocycles such as
imidazole, hydrazino or hydroxylamino groups, isocyanate or cyanate
groups, and sulfur containing groups such as sulfoxide, sulfone,
sulfide, and disulfide. Additionally, the substitutions can be via
single, double, or triple bonds, when relevant or possible.
[0039] Further, aliphatic groups may also contain hetero
substitutions, which are substitutions of carbon atoms, by hetero
atoms such as, for example, nitrogen, oxygen, phosphorous, or
sulfur. As such, a linker comprised of a substituted aliphatic can
have a backbone comprised of carbon, nitrogen, oxygen, sulfur,
phosphorous, and/or the like. Heterocyclic substitutions refer to
alkyl rings having one or more hetero atoms.
[0040] Examples of heterocyclic moieties include but are not
limited to morpholino, imidazole, tetrahydrofuran, and
pyrrolidino.
[0041] As used herein, the term "aryl" or "aromatic" is meant to
refer to molecule is one in which electrons are free to cycle
around circular or cyclic arrangements of atoms, which are
alternately singly and doubly bonded to one another. More properly,
these bonds may be seen as a hybrid of a single bond and a double
bond, each bond in the ring being identical to every other.
Examples of aromatic compounds that can be present include benzene,
benzyl, toluene, xylene, and the like. The aromatic compound can
include hetero atoms so as to be a hetero aromatic such as
pyridine, furan, and the like. Also, an aromatic can be a
polycyclic aromatic such as naphthalene, anthracene, phenanthrene,
polycyclic aromatic hydrocarbons, indole, quinoline, isoquinoline,
and the like. Any aryl herein can be a heteroaryl or polyaryl.
[0042] As used herein, the term "amine" is meant to refer to
moieties that can be derived directly or indirectly from ammonia by
replacing one, two, or three hydrogen atoms by other groups, such
as, for example, alkyl groups. Primary amines have the general
structures RNH.sub.2 and secondary amines have the general
structure R.sub.2NH, where R can be any of R1-R5. The term amine
includes, but is not limited to methylamine, ethylamine,
propylamine, isopropylamine, aniline, cyclohexylamine, benzylamine,
polycyclic amines, heteroatom substituted aryl and alkylamines,
dimethylamine, diethylamine, diisopropylamine, dibutylamine,
methylpropylamine, methylhexylamine, methylcyclopropylamine,
ethylcylohexylamine, methylbenzylamine, methycyclohexylmethylamine,
butylcyclohexylamine, morpholine, thiomorpholine, pyrrolidine,
piperidine, 2,6-dimethylpiperidine, piperazine, and heteroatom
substituted alkyl or aryl secondary amines.
[0043] As used herein, the term "halo" means fluoro, chloro, bromo,
or iodo, preferably fluoro and chloro.
[0044] As used herein, the term "poly(amino acid)" or "polypeptide"
is a polyamide formed from amino acids. Poly(amino acid)s will
generally range from about 200-2,000 molecular weight or greater
than about 2,000 molecular weight, or having no upper molecular
weight limit, and normally being less than 10,000,000 and usually
not more than about 600,000 daltons. The amino acids can be
natural, unnatural, common, essential, non-essential or analogs or
derivatives thereof.
[0045] As used herein, the term "peptide" is meant to refer to any
compound formed by the linkage of two or more amino acids by amide
(peptide) bonds, usually a polymer of .alpha.-amino acids in which
.alpha.-amino group of each amino acid residue (except the NH.sub.2
terminus) is linked to the .alpha.-carboxyl group of the next
residue in a linear chain. The terms "peptide," "polypeptide," and
"poly(amino acid)" are used synonymously herein to refer to this
class of compounds without restriction as to size. The largest
members of this class are referred to as proteins.
[0046] Additionally, some of the compounds of the present invention
can be prepared as racemic mixtures of isomers, mixtures of
isomers, or optically isolated isomers. Compounds that have the
same molecular formula but differ in the nature or sequence of
bonding of their atoms or the arrangement of their atoms in space
are termed "isomers." Isomers that differ in the arrangement of
their atoms in space are termed "stereoisomers."
[0047] Stereoisomers that are not mirror images of one another are
termed "diastereomers" and those that are non-superimposable mirror
images of each other are termed "enantiomers". When a compound has
an asymmetric center, for example, it is bonded to four different
groups, a pair of enantiomers is possible. An enantiomer can be
characterized by the absolute configuration of its asymmetric
center and is described by the R- and S-sequencing rules of Cahn
and Prelog, or by the manner in which the molecule rotates the
plane of polarized light and designated as dextrorotatory or
levorotatory (i.e., as (+) or (-)-isomers respectively). A chiral
compound can exist as either individual enantiomer or as a mixture
thereof. A mixture containing equal proportions of the enantiomers
is called a "racemic mixture". The present invention can include
racemic mixtures of the compounds defined by Formulas 1-15 and the
pure individual enantiomers. All possible enantiomers under
Formulas 1-15 are considered to be disclosed herein.
[0048] The compounds of this invention may possess one or more
asymmetric centers. Unless indicated otherwise, the description or
naming of a particular compound in the specification and claims is
intended to include both individual enantiomers and mixtures,
racemic or otherwise, thereof. The methods for the determination of
stereochemistry and the separation of stereoisomers are well-known
in the art (see discussion in Chapter 4 of "Advanced Organic
Chemistry", 4.sup.th edition J. March, John Wiley and Sons, New
York, 1992).
[0049] In some embodiments, the compounds in the compositions may
be present as a pharmaceutically acceptable salt. The
pharmaceutically acceptable salts includes salts of the active
agent or components of the composition, prepared, for example, with
acids or bases, depending on the particular substituents found
within the composition and the treatment modality desired.
Pharmaceutically acceptable salts can be prepared as alkaline metal
salts, such as lithium, sodium, or potassium salts; or as alkaline
earth salts, such as beryllium, magnesium or calcium salts.
Examples of suitable bases that may be used to form salts include
ammonium, or mineral bases such as sodium hydroxide, lithium
hydroxide, potassium hydroxide, calcium hydroxide, magnesium
hydroxide, and the like. Examples of suitable acids that may be
used to form salts include inorganic or mineral acids such as
hydrochloric, hydrobromic, hydroiodic, hydrofluoric, nitric,
carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric,
dihydrogenphosphoric, sulfuric, monohydrogensulfuric, phosphorous
acids and the like. Other suitable acids include organic acids, for
example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric
acid, phosphoric acid and the like, and organic acids such as
trifluoroacetic, acetic, propionic, glycolic, pyruvic, oxalic,
maleic, malonic, succinic, fumaric, tartaric, citric, benzoic,
cinnamic, mandelic, methanesulfonic, ethanesulfonic,
p-toluenesulfonic, salicylic, isobutyric, suberic, phthalic,
benzenesulfonic, p-tolylsulfonic, salicylic, formic,
naphthalene-2-sulfonic, and the like. Still other suitable acids
include amino acids such as arginate, aspartate, glutamate, and the
like.
[0050] Additionally, the compounds can be prepared to be prodrugs
that include a cleavable linker between the base analog and the
prodrug portion. Phosphate groups that cleave to leave hydroxyl
groups are one example of produg moieties. Also, the ester
compounds described herein can be cleaved in vivo and they
themselves may be considered prodrugs. The prodrugs, such as the
esters, can have shorter half lives while retaining opioid receptor
activity.
[0051] As used herein, the term "analog" or the like is meant to
refer to a structurally related compound or compounds with a common
scaffold that different functional groups or substituents. For
example, the different R groups in Formulas 1-14 can be prepared
into analogs of each other by changing one or more R groups.
[0052] As used herein, the term "derivative" or the like is meant
to refer to a substitution of an atom with another atom or group.
For example, when a hydrogen is replaced with a halogen or an alkyl
group, such a change produces a derivative.
Pharmaceutical Compositions and Methods
[0053] Generally, the pharmaceutical compositions can be used for
providing a compound in an effective amount for interacting with an
opioid receptor (OR). Such a composition can include a
pharmaceutically acceptable carrier containing a functionalized
polycycle analog as described herein, such as an analog or
derivative of the chemical structures shown herein and described in
the tables. The compounds can be analogs of Formulas 1-16. It is
surprising and unexpected that the compounds described herein of
Formulas 1-16 can modulate opioid receptors because these compounds
have substantially no similarity to other compounds, such as
opiates, that modulate opioid compounds.
[0054] The compound can be present in a therapeutically effective
amount for providing any function for any opioid receptor at any
location. Examples include the following: providing analgesia;
providing anorectic characteristics; providing weight loss;
treating, inhibiting, and/or preventing depression; treating,
inhibiting, and/or preventing a mood disorder; treating,
inhibiting, and/or preventing bipolar disorders; treating,
inhibiting, and/or preventing drug addiction; treating, inhibiting,
and/or preventing drug seeking behavior; or treating, inhibiting,
and/or preventing stress-induced drug seeking behavior; treating,
inhibiting, and/or preventing inflammatory disorders; treating,
inhibiting, and/or preventing renal disorders; treating,
inhibiting; and/or preventing cardiovascular disorders; psychotic
disorders. Additionally, the therapeutically effective amount can
be an amount sufficient to provide a therapeutic benefit associated
with agonizing or antagonizing an opioid receptor, such as the
activities of Table A.
[0055] The compounds can be used for modulating opioid receptors
present in human or animal tissue in vitro or in vivo. This can
include administering an effective amount of a compound that is an
opioid receptor agonist or antagonist to a subject such that a
sufficient amount of the compound active in the brain for
modulating opioid receptor activity. For example, the
administration can be via subcutaneous, intravenous, inhalation,
and the like.
[0056] In one embodiment, the compound can be selective for a
specific opioid receptor and preferentially target a DOR, KOR, MOR
or NR. The compound can be more effective at modulating one opioid
receptor over another receptor. Moreover, the compound can be
capable of crossing the blood brain barrier. Also, the compound may
be insufficient for substantial interaction with other opioid
receptors
[0057] The compounds of the present invention can be formulated
into a pharmaceutically acceptable formulation. Such a composition
can be useful to prevent, alleviate, eliminate, inhibit or delay
the onset of a disease, disorder, and/or condition related thereto.
Accordingly pharmaceutical compositions can be used as a
prophylactic or treatment for a disease, disorder, and/or
condition.
[0058] In embodiments of the present invention, the pharmaceutical
composition comprises at least one active component and inactive
components. The active components are an opioid receptor modulation
compound as described herein and their derivatives/analogues,
salts, and prodrugs thereof. The inactive components are selected
from the group consisting of excipients, carriers, solvents,
diluents, stabilizers, enhancers, additives, adhesives, and
combinations thereof.
[0059] The amount of the compound in a formulation can vary within
the full range employed by those skilled in the art. Typically, the
formulation will contain, on a weight percent basis, from about
0.01-99.99 weight percent of the compounds of the present invention
based on the total formulation, with the balance being one or more
suitable pharmaceutical excipients. Preferably, the compounds are
present at a level of about 1-80 weight percent.
[0060] Pharmaceutical preparations include sterile aqueous or
non-aqueous solutions, suspensions and emulsions. Examples of
non-aqueous solvents are propylene glycol, polyethylene glycol,
vegetable oil such as olive oil, injectable organic esters such as
ethyloliate. Aqueous carriers include water, alcoholic/aqueous
solutions, emulsions or suspensions, including saline and buffered
media. Parenteral vehicles include sodium chloride solution,
Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's
or fixed oils. Intravenous vehicles include fluid and nutrient
replenishers, electrolyte replenishers, (such as those based on
Ringer's dextrose), and the like. Preservatives and other additives
may also be present such as, for example, antimicrobials,
antioxidants, chelating agents and inert gases and the like. Those
of skill in the art can readily determine the various parameters
for preparing these pharmaceutical compositions without resort to
undue experimentation.
[0061] Pharmacological compositions may be prepared from
water-insoluble compounds, or salts thereof, such as aqueous base
emulsions. In such embodiments, the pharmacological composition
will typically contain a sufficient amount of pharmaceutically
acceptable emulsifying agent to emulsify the desired amount of the
pharmacological agent. Useful emulsifying agents include, but are
not limited to, phosphatidyl cholines, lecithin, and the like.
[0062] Additionally, the compositions may contain other additives,
such as pH-adjusting additives. In particular, useful pH-adjusting
agents include acids, such as hydrochloric acid, bases or buffers,
such as sodium lactate, sodium acetate, sodium phosphate, sodium
citrate, sodium borate, or sodium gluconate. Furthermore,
pharmacological agent compositions may, though not always, contain
microbial preservatives. Microbial preservatives that may be
employed include, but are not limited to, methylparaben,
propylparaben, and benzyl alcohol. The microbial preservative may
be employed when the pharmacological agent formulation is placed in
a vial designed for multi-dose use. Pharmacological agent
compositions for use in practicing the subject methods may be
lyophilized using techniques well known in the art.
[0063] The pharmaceutically acceptable excipients, such as
vehicles, adjuvants, carriers or diluents, are readily available to
the public. Examples of suitable excipients can include, but are
not limited to, the following: acidulents, such as lactic acid,
hydrochloric acid, and tartaric acid; solubilizing components, such
as non-ionic, cationic, and anionic surfactants; absorbents, such
as bentonite, cellulose, and kaolin; alkalizing components, such as
diethanolamine, potassium citrate, and sodium bicarbonate;
anticaking components, such as calcium phosphate tribasic,
magnesium trisilicate, and talc; antimicrobial components, such as
benzoic acid, sorbic acid, benzyl alcohol, benzethonium chloride,
bronopol, alkyl parabens, cetrimide, phenol, phenylmercuric
acetate, thimerosol, and phenoxyethanol; antioxidants, such as
ascorbic acid, alpha tocopherol, propyl gallate, and sodium
metabisulfite; binders, such as acacia, alginic acid, carboxymethyl
cellulose, hydroxyethyl cellulose; dextrin, gelatin, guar gum,
magnesium aluminum silicate, maltodextrin, povidone, starch,
vegetable oil, and zein; buffering components, such as sodium
phosphate, malic acid, and potassium citrate; chelating components,
such as EDTA, malic acid, and maltol; coating components, such as
adjunct sugar, cetyl alcohol, polyvinyl alcohol, carnauba wax,
lactose maltitol, titanium dioxide; controlled release vehicles,
such as microcrystalline wax, white wax, and yellow wax;
desiccants, such as calcium sulfate; detergents, such as sodium
lauryl sulfate; diluents, such as calcium phosphate, sorbitol,
starch, talc, lactitol, polymethacrylates, sodium chloride, and
glyceryl palmitostearate; disintegrants, such as colloidal silicon
dioxide, croscarmellose sodium, magnesium aluminum silicate,
potassium polacrilin, and sodium starch glycolate; dispersing
components, such as poloxamer 386, and polyoxyethylene fatty esters
(polysorbates); emollients, such as cetearyl alcohol, lanolin,
mineral oil, petrolatum, cholesterol, isopropyl myristate, and
lecithin; emulsifying components, such as anionic emulsifying wax,
monoethanolamine, and medium chain triglycerides; flavoring
components, such as ethyl maltol, ethyl vanillin, fumaric acid,
malic acid, maltol, and menthol; humectants, such as glycerin,
propylene glycol, sorbitol, and triacetin; lubricants, such as
calcium stearate, canola oil, glyceryl palmitostearate, magnesium
oxide, poloxymer, sodium benzoate, stearic acid, and zinc stearate;
solvents, such as alcohols, benzyl phenylformate, vegetable oils,
diethyl phthalate, ethyl oleate, glycerol, glycofurol, for indigo
carmine, polyethylene glycol, for sunset yellow, for tartazine,
triacetin; stabilizing components, such as cyclodextrins, albumin,
xanthan gum; and tonicity components, such as glycerol, dextrose,
potassium chloride, and sodium chloride; and mixture thereof.
Excipients include those that alter the rate of absorption,
bioavailability, or other pharmacokinetic properties of
pharmaceuticals, dietary supplements, alternative medicines, or
nutraceuticals.
[0064] Other examples of suitable excipients, binders and fillers
are listed in Remington's Pharmaceutical Sciences, 18th Edition,
ed. Alfonso Gennaro, Mack Publishing Co. Easton, Pa., 1995 and
Handbook of Pharmaceutical Excipients, 3rd Edition, ed. Arthur H.
Kibbe, American Pharmaceutical Association, Washington D.C. 2000,
both of which are incorporated herein by reference.
[0065] In general, pharmaceutically acceptable carriers for are
well-known to those of ordinary skill in the art. This carrier can
be a solid or liquid and the type is generally chosen based on the
type of administration being used. Suitable pharmaceutical carriers
are, in particular, fillers, such as sugars, for example lactose,
sucrose, mannitol or sorbitol, cellulose preparations and/or
calcium phosphates, for example tricalcium phosphate or calcium
hydrogen phosphate, furthermore, binders such as starch paste,
using, for example, corn, wheat, rice or potato starch, gelatin,
tragacanth, methylcellulose and/or polyvinylpyrrolidone, if
desired, disintegrants, such as the above mentioned starches;
furthermore carboxymethyl starch, crosslinked polyvinylpyrrolidone,
agar, alginic acid or a salt thereof, such as sodium alginate;
auxiliaries are primarily glidants, flow-regulators and lubricants,
for example silicic acid, talc, stearic acid or salts thereof, such
as magnesium or calcium stearate, and/or polyethylene glycol.
Sugar-coated tablet cores are provided with suitable coatings
which, if desired, are resistant to gastric juice, using, inter
alia, concentrated sugar solutions which, if desired, contain gum
arabic, talc, polyvinylpyrrolidone, polyethylene glycol and/or
titanium dioxide, coating solutions in suitable organic solvents or
solvent mixtures or, for the preparation of gastric juice-resistant
coatings, solutions of suitable cellulose preparations, such as
acetylcellulose phthalate or hydroxypropylmethylcellulose
phthalate. Colorants or pigments, for example, to identify or to
indicate different doses of active ingredient, may be added to the
tablets or sugar-coated tablet coatings.
[0066] Additional pharmaceutically acceptable carriers that may be
used in these pharmaceutical compositions include, but are not
limited to, ion exchangers, alumina, aluminum stearate, lecithin,
serum proteins, such as human serum albumin, buffer substances such
as phosphates, glycine, sorbic acid, potassium sorbate, partial
glyceride mixtures of saturated vegetable fatty acids, water, salts
or electrolytes, such as prolamine sulfate, disodium hydrogen
phosphate, potassium hydrogen phosphate, sodium chloride, zinc
salts, colloidal silica, magnesium trisilicate, polyvinyl
pyrrolidone, cellulose-based substances, polyethylene glycol,
sodium carboxymethylcellulose, polyacrylates, waxes,
polyethylene-polyoxypropylene-block polymers, polyethylene glycol
and wool fat.
[0067] Additional formulations for use in the present invention can
be found in Remington's Pharmaceutical Sciences (Mack Publishing
Company, Philadelphia, Pa., 17th ed. (1985)), which is incorporated
herein by reference. Moreover, for a brief review of methods for
drug delivery, see, Langer, Science 249:1527-1533 (1990), which is
incorporated herein by reference. The pharmaceutical compositions
described herein can be manufactured in a manner that is known to
those of skill in the art, i.e., by means of conventional mixing,
dissolving, granulating, dragee-making, levigating, emulsifying,
encapsulating, entrapping or lyophilizing processes. Other examples
of suitable pharmaceuticals are listed in 2000 Med Ad News 19:56-60
and The Physicians Desk Reference, 53rd edition, 792-796, Medical
Economics Company (1999), both of which are incorporated herein by
reference.
[0068] In general, compounds of this invention can be administered
as pharmaceutical compositions by any one of the following routes:
oral, systemic (e.g., transdermal, intranasal or by suppository),
or parenteral (e.g., intramuscular, intravenous or subcutaneous)
administration. One manner of administration is oral using a
convenient daily dosage regimen which can be adjusted according to
the degree of affliction. Compositions can take the form of
tablets, pills, capsules, semisolids, powders, sustained release
formulations, solutions, suspensions, elixirs, aerosols, or any
other appropriate compositions. Another manner for administering
compounds of this invention is inhalation.
[0069] According to the methods of the present invention, the
compositions of the invention can be administered by injection by
gradual infusion over time or by any other medically acceptable
mode. Any medically acceptable method may be used to administer the
composition to the patient. The particular mode selected will
depend of course, upon factors such as the particular drug
selected, the severity of the state of the subject being treated,
or the dosage required for therapeutic efficacy. The methods of
this invention, generally speaking, may be practiced using any mode
of administration that is medically acceptable, meaning any mode
that produces effective levels of the active composition without
causing clinically unacceptable adverse effects.
[0070] The administration may be localized (i.e., to a particular
region, physiological system, tissue, organ, or cell type) or
systemic. For example, the composition may be administered through
parental injection, implantation, orally, vaginally, rectally,
buccally, pulmonary, topically, nasally, transdermally, surgical
administration, or any other method of administration where access
to the target by the composition is achieved. In one example, the
administration is directly into the brain or brain cavity. Examples
of parenteral modalities that can be used with the invention
include intravenous, intradermal, subcutaneous, intracavity,
intramuscular, intraperitoneal, epidural, or intrathecal. Examples
of implantation modalities include any implantable or injectable
drug delivery system. Oral administration may be used for some
treatments because of the convenience to the patient as well as the
dosing schedule. Compositions suitable for oral administration may
be presented as discrete units such as capsules, pills, cachettes,
tables, or lozenges, each containing a predetermined amount of the
active compound. Other oral compositions include suspensions in
aqueous or non-aqueous liquids such as a syrup, an elixir, or an
emulsion.
[0071] The compounds can be encapsulated in a vehicle such as
liposomes that facilitates transfer of the bioactive molecules into
the targeted tissue, as described, for example, in U.S. Pat. No.
5,879,713 to Roth et al. and Woodle, et al., U.S. Pat. No.
5,013,556, the contents of which are hereby incorporated by
reference. The compounds can be targeted by selecting an
encapsulating medium of an appropriate size such that the medium
delivers the molecules to a particular target. For example,
encapsulating the compounds within microparticles, preferably
biocompatible and/or biodegradable microparticles, which are
appropriate sized to infiltrate, but remain trapped within, the
capillary beds and alveoli of the lungs can be used for targeted
delivery to these regions of the body following administration to a
patient by infusion or injection.
[0072] Microparticles can be fabricated from different polymers
using a variety of different methods known to those skilled in the
art. The solvent evaporation technique is described, for example,
in E. Mathiowitz, et al., J. Scanning Microscopy, 4, 329 (1990); L.
R. Beck, et al., Fertil. Steril., 31, 545 (1979); and S. Benita, et
al., J. Pharm. Sci., 73, 1721 (1984). The hot-melt
microencapsulation technique is described by E. Mathiowitz, et al.,
Reactive Polymers, 6, 275 (1987). The spray drying technique is
also well known to those of skill in the art. Spray drying involves
dissolving a suitable polymer in an appropriate solvent. A known
amount of the compound is suspended (insoluble drugs) or
co-dissolved (soluble drugs) in the polymer solution. The solution
or the dispersion is then spray-dried. Microparticles ranging
between 1-10 microns are obtained with a morphology which depends
on the type of polymer used.
[0073] Embodiments may also include administration of at least one
pharmacological agent using a pharmacological delivery device or
system such as, but not limited to, pumps (implantable or external
devices), epidural injectors, syringes or other injection
apparatus, catheter and/or reservoir operatively associated with a
catheter, injection, and the like. For example, in certain
embodiments a delivery device employed to deliver at least one
pharmacological agent to a subject may be a pump, syringe, catheter
or reservoir operably associated with a connecting device such as a
catheter, tubing, or the like. Containers suitable for delivery of
at least one pharmacological agent to a pharmacological agent
administration device include instruments of containment that may
be used to deliver, place, attach, and/or insert at least one
pharmacological agent into the delivery device for administration
of the pharmacological agent to a subject and include, but are not
limited to, vials, ampules, tubes, capsules, bottles, syringes and
bags. In one embodiment, a catheter can be used to direct the
composition directly to the brain or other location in the body for
systemic delivery.
[0074] The compositions of the present invention may be given in
dosages, generally at the maximum amount while avoiding or
minimizing any potentially detrimental side effects. The
compositions can be administered in effective amounts, alone or in
a cocktail with other compounds, for example, other compounds that
can be used to treat, inhibit, or prevent drug addiction or
drug-seeking behavior.
[0075] In one embodiment of the present invention, therapeutically
effective amounts of compounds of the present invention may range
from approximately 0.05 to 50 mg per kilogram body weight of the
recipient per day; preferably about 0.01-25 mg/kg/day, more
preferably from about 0.5 to 10 mg/kg/day. Thus, for administration
to a 70 kg person, the dosage range would most preferably be about
35-70 mg per day.
[0076] In another embodiment of the present invention, dosages may
be estimated based on the results of experimental models,
optionally in combination with the results of assays of the present
invention. Generally, daily oral doses of active compounds will be
from about 0.01 mg/kg per day to 2000 mg/kg per day. Oral doses in
the range of 10 to 500 mg/kg, in one or several administrations per
day, may yield suitable results. In the event that the response of
a particular subject is insufficient at such doses, even higher
doses (or effective higher doses by a different, more localized
delivery route) may be employed to the extent that patient
tolerance permits. Multiple doses per day are also contemplated in
some cases to achieve appropriate systemic levels of the
composition. Use of a long-term release implant may be particularly
suitable in some cases. "Long-term release," as used herein, means
that the implant is constructed and arranged to deliver therapeutic
levels of the composition for at least 30 or 45 days, and
preferably at least 60 or 90 days, or even longer in some cases.
Long-term release implants are well known to those of ordinary
skill in the art, and include some of the release systems described
above.
[0077] Dosage amount and interval may be adjusted individually to
provide plasma levels of the active compound which are sufficient
to maintain therapeutic effect. Preferably, therapeutically
effective serum levels will be achieved by administering multiple
doses each day. In cases of local administration or selective
uptake, the effective local concentration of the drug may not be
related to plasma concentration. One having skill in the art will
be able to optimize therapeutically effective local dosages without
undue experimentation.
Synthesis
[0078] Generally, the methods for synthesizing a functionalized
polycyclic analog, such as those having a carboxylic acid, amide,
and/or ester can include one or more reactions in accordance with
one or more of Schemes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and/or
combinations thereof. This can include synthesizing one or a
plurality of functionalized polycyclic analog, such as those
according to Formulas 1-15.
[0079] In one embodiment, the invention includes a method for
preparing a functionalized polycyclic compound having a carboxylic
acid. Such a method can include: providing a hydroxyl-containing
diene or an amine-containing diene; and reacting the diene with a
dienophile under sufficient conditions for a combined
Diels-Alder/acylation reaction so as to provide a polycyclic
compound having a carboxylic acid having Formula 17:
##STR00006##
In Formula 17, R3, R4, R5, and R6 are independently a hydrogen,
halogen, hydroxyl, straight or branched substituted or
unsubstituted alkoxy, amine, straight or branched substituted or
unsubstituted alkyl, substituted or unsubstituted cycloalkyl,
substituted or unsubstituted aryl, branched or unbranched or cyclic
substituted or unsubstituted arylalkyl, or combinations thereof, or
R4 and R5 together, form a ring therebetween; X is an O, N, or S;
and n is from 0 to 5. R3-R6 can be as described herein.
[0080] Additionally, the compounds herein of Formulas 1-16 can be
prepared in accordance with the compound of Formula 17, and then
further reacted by standard chemical synthesis to obtain any of the
compounds of Formulas 1-16. While the specific protocol is
described for Formula 17, the compounds can undergo further
reaction to prepare the other compounds. Also, the starting
reagents can be modulated in order to have the various R1-R18
groups as shown in Formula 1.
[0081] In one embodiment, the reaction can include preparing an
amine-containing diene component. For example, the amine-containing
diene component is prepared from 3,5-hexadien-1-ol.
[0082] In one embodiment, the reaction can include a thermal
reaction of a diene with a dienophile such as a maleic anhydride.
For example, when the thermal reaction provides a functionalized
polycyclic carboxylic acid analog. The reaction can be in a solvent
(e.g., dichloroethane or toluene) or neat. The thermal reaction
conditions can include a temperature from about 25.degree. C. to
about 165.degree. C., more preferably about 100.degree. C. to about
165.degree. C., and most preferably from about 150.degree. C. to
about 165.degree. C. The thermal reaction conditions can be
conducted for a duration from about 1 min to about 1.5 hours, more
preferably about 30 minutes to about 1.5 hours, and most preferably
from about 30 minutes to about 1 hour.
[0083] In one embodiment, a synthetic method can also include
separation of individual enantiomers from a racemic mixture into
compositions having a substantially pure enantiomer. The purified
enantiomers can be considered to be more pure for a specific
enantiomer compared to the racemic mixture.
[0084] The synthetic methods have been used to prepare heterocyclic
libraries having potential biological activity. A Diels-Alder
reaction was modified so that a reaction of maleic anhydride
dienophile with an amine-containing diene was conducted as shown in
Scheme 1. The reaction can produce an amine-containing side chain
that is cis to a reactive carbonyl group. This provides a
carboxylic acid group for downstream manipulation.
##STR00007##
[0085] The synthesis of the amine-containing diene components can
be conducted from 3,5-hexadien-1-ol, which is readily synthesized
from ethyl sorbate by deconjugation and reduction. Other
alkyldiene-ols can also be used to prepare the amine-containing
diene. Mesylation and subsequent displacement with a primary amine
readily produced the desired aminodienes in reasonable overall
yields and on 1-2 g scale (Table 1). The displacement of mesylate
by the amine was facilitated by microwave irradiation
(acetonitrile, 130.degree. C., 1 h); however, other methods to
increase the temperature can be used. The amines were purified by
silica gel chromatography prior to use in the next step. The
compound number is identified by C#.
TABLE-US-00002 TABLE 1 Synthesis of the 1-amino-3,5-hexadienes
1{1-6}. ##STR00008## ##STR00009## entry R.sub.1 C# yield (%) 1
n-butyl 1{1} 87 2 cyclopropyl 1{2} 40 3 cyclohexyl 1{3} 78 4 benzyl
1{4} 92 5 3,4-dichlorobenzyl 1{5} 97 6 3,4-dimethoxybenzyl 1{6}
80
[0086] The thermal reactions of diene reagents 1(1-6) and maleic
anhydride were also studied through Table 2, as shown below. High
internal pressures and temperatures were obtained by microwave
irradiation, which facilitated the overall reaction. The diene
reagents (1(1-6)) and maleic anhydride were combined in a 10 mL
microwave vial in dichloroethane (DCE) and heated to 165.degree. C.
After 1.5 h, maximum yields of the functionalized polycyclic
analogs 2(1-6) were obtained. A survey of reaction conditions
showed that improved yields were obtained using 1.25 equivalents of
the dienophile (maleic anhydride) relative to the diene 1(1-6).
While various organic solvents can be used in the process, such as
CH.sub.2Cl.sub.2, toluene, and CH.sub.3CN, dichloroethane obtained
improved conversion in this reaction. In addition, the relatively
high temperatures and ca. 1.5 h reaction times were also found to
improve or provide optimal yields.
[0087] The synthetic protocol was initially conducted with six
dienes being reacted with maleic anhydride to produce a series of
six functionalized polycyclic scaffolds (Table 2). All of the
reactions shown gave functionalized polycyclic scaffolds in good
yields and, when carried out on scale, in 0.5-1.5 g quantities.
TABLE-US-00003 TABLE 2 Diels-Alder Reactions of Dienes 1{1-6} with
Maleic Anhydride. ##STR00010## ##STR00011## entry diene R.sub.1
product yield (%) 1 1{1} n-butyl 2{1} 74 2 1{2} cyclopropyl 2{2} 76
3 1{3} cyclohexyl 2{3} 68 4 1{4} benzyl 2{4} 74 5 1{5}
3,4-dichlorobenzyl 2{5} 80 6 1{6} 3,4-dimethoxybenzyl 2{6} 80
[0088] The modified reaction can be conducted with other
amino-containing dienes that may have a longer or shorter alkyl
group as well as an alkyl group that is straight or branched
substituted or unsubstituted. The maleic anhydride dienophile may
also be replaced with other dienophiles.
[0089] Two additional dienophiles were shown to produce
cycloaddition products. The reaction of an aminodiene 1(2) with
citraconic anhydride under the above conditions produced a methyl
functionalized polycyclic product 3(2) in 54% yield (Scheme 3). In
addition, the reaction of an aminodiene 1(5) and citraconic
anhydride at 100.degree. C. thermal heating for 5.5 hours without
solvent gave the functionalized polycyclic product 3(5) in 76%
yield (Scheme 2).
##STR00012##
[0090] Additionally, dimethyl fumarate (it may be substituted with
another dialkyl fumarate) was reacted with aminodiene 1(5) and to
give compounds 4a and 4b, as an equimolar mixture of isomers, in
68-76% combined yield (Scheme 3). Treatment of adduct 2(5) with
(trimethylsilyl)diazomethane smoothly produced the ester 5, which
was shown to be isomerically distinct from 4a and 4b (Scheme 3),
demonstrating that both Diels-Alder reactions were
stereoselective.
##STR00013##
Library Synthesis
[0091] The methods of chemical synthesis can be used to prepare a
scaffold having a free carboxylic acid group that can be reacted
with various amines to produce a library of analogs. For example,
the amine can be any of structures 6(1) through 6(12) or related
amines or other amines.
##STR00014## ##STR00015##
[0092] In one embodiment, the functionalizing reaction between the
polycyclic carboxylic acid and an amine can be catalyzed with a
catalyst. For example, the catalyst can be DMAP and/or the coupling
reagent EDC-HCl).
[0093] The six scaffolds 2(1-6) prepared as described were
subjected to an additional diversity step to prepare analogs. The
carboxylic acids were reacted with the twelve amines 6(1-12) shown
above using a catalytic amount of DMAP and
N-(3'-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride
(EDC.HCl) as the primary coupling reagent (Scheme 4). The amine
components (6(1-12)) could be substituted with other amines to
produce different analogs. The examples show productive coupling of
the functionalized polycyclic analog scaffolds over a range of
amines with diverse chemical reactivity. The reactions were stirred
at room temperature for 14 h then partitioned between
CH.sub.2Cl.sub.2 and water in phase separator tubes fitted with
hydrophobic filters. The organic layers obtained were directly
subjected to solid phase extraction (SPE). Elution with
CH.sub.2Cl.sub.2:acetone (1:1) provided the crude amide coupled
products. The compounds thus prepared were subjected to
mass-directed HPLC purification to produce the adducts shown (Table
3). The compounds can be identified by combining the functionalized
polycyclic analog scaffold and the amine groups to form an
amide.
##STR00016##
TABLE-US-00004 TABLE 3 Parallel synthesis of a 72-member
functionalized polycyclic amide library. HRMS, crude final calcd
scaf- purity purified purity for HRMS, product fold amine (%) yield
(%) [M + H].sup.+ found 7{1} 2{1} 6{1} 97 26 mg 100 321.2178
321.2186 (82%) 7{2} 2{1} 6{2} 88 35 mg 100 396.2651 396.2655 (88%)
7{3} 2{1} 6{3} 86 31 mg 91 355.2386 355.2395 (88%) 7{4} 2{1} 6{4}
91 21 mg 100 307.2386 307.2399 (68%) 7{5} 2{1} 6{5} 93 21 mg 100
333.2542 333.2568 (62%) 7{6} 2{1} 6{6} 84 31 mg 100 371.2335
371.2359 (85%) 7{7} 2{1} 6{7} 93 11 mg 100 342.2182 342.2205 (31%)
7{8} 2{1} 6{8} 91 28 mg 98 334.1589 334.1610 (84%) 7{9} 2{1} 6{9}
84 25 mg 100 327.2073 327.2091 (78%) 7{10} 2{1} 6{10} 81 31 mg 100
357.2178 357.2211 (87%) 7{11} 2{1} 6{11} 60 29 mg 100 395.1293
395.1315 (73%) 7{12} 2{1} 6{12} 71 32 mg 99 429.1557 429.1585 (75%)
7{13} 2{2} 6{1} 80 18 mg 87.sup.a 305.1865 305.1884 (60%).sup.a
7{14} 2{2} 6{2} 79 26 mg 91 380.2338 380.2351 (69%) 7{15} 2{2} 6{3}
84 17 mg 97 339.2073 339.2079 (50%) 7{16} 2{2} 6{4} 89 20 mg 100
291.2073 291.2088 (67%) 7{17} 2{2} 6{5} 10 1 mg 100 317.2229
317.2252 (6%) 7{18} 2{2} 6{6} 97 22 mg 97 355.2022 355.2034 (61%)
7{19} 2{2} 6{7} 84 2 mg 100 326.1869 326.1887 (7%) 7{20} 2{2} 6{8}
54 18 mg 99 318.1276 318.1291 (57%) 7{21} 2{2} 6{9} 85 13 mg 100
311.1760 311.1784 (42%) 7{22} 2{2} 6{10} 74 19 mg 100 341.1865
341.1890 (56%) 7{23} 2{2} 6{11} 50 21 mg 99 379.0980 379.0995 (56%)
7{24} 2{2} 6{12} 57 29 mg 100 413.1244 413.1268 (70%) 7{25} 2{3}
6{1} 95 18 mg 100 347.2335 347.2345 (53%) 7{26} 2{3} 6{2} 89 28 mg
99 422.2808 422.2815 (65%) 7{27} 2{3} 6{3} 88 24 mg 99 381.2542
381.2547 (63%) 7{28} 2{3} 6{4} 91 14 mg 100 333.2542 333.2550 (42%)
7{29} 2{3} 6{5} 85 30 mg 96 359.2699 359.2714 (83%) 7{30} 2{3} 6{6}
65 36 mg 98 397.2491 397.2509 (90%) 7{31} 2{3} 6{7} 99 18 mg 95
368.2338 368.2372 (50%) 7{32} 2{3} 6{8} 77 20 mg 95 360.1746
360.1762 (56%) 7{33} 2{3} 6{9} 86 24 mg 98 353.2229 353.2241 (69%)
7{34} 2{3} 6{10} 65 30 mg 98 383.2335 383.2347 (78%) 7{35} 2{3}
6{11} 49 22 mg 100 421.1450 421.1469 (51%) 7{36} 2{3} 6{12} 69 32
mg 99 455.1713 455.1721 (69%) 7{37} 2{4} 6{1} 89 21 mg 100 355.2022
355.2035 (59%) 7{38} 2{4} 6{2} 87 31 mg 97 430.2495 430.2491 (72%)
7{39} 2{4} 6{3} 88 29 mg 96 389.2229 389.2237 (75%) 7{40} 2{4} 6{4}
82 14 mg 98 341.2229 341.2236 (42%) 7{41} 2{4} 6{5} 65 30 mg 100
367.2386 367.2401 (81%) 7{42} 2{4} 6{6} 76 39 mg 99 405.2178
405.2204 (96%) 7{43} 2{4} 6{7} 99 20 mg 97 376.2025 376.2053 (54%)
7{44} 2{4} 6{8} 97 32 mg 99 368.1433 368.1456 (90%) 7{45} 2{4} 6{9}
91 34 mg 100 361.1916 361.1924 (93%) 7{46} 2{4} 6{10} 87 35 mg 100
391.2022 391.2031 (85%) 7{47} 2{4} 6{11} 28 11 mg 100 429.1137
429.1150 (26%) 7{48} 2{4} 6{12} 72 45 mg 100 463.1400 463.1426
(99%) 7{49} 2{5} 6{1} 98 21 mg 100 423.1242 423.1257 (48%) 7{50}
2{5} 6{2} 84 29 mg 99 498.1715 498.1718 (58%) 7{51} 2{5} 6{3} 82 23
mg 99 457.1450 457.1463 (49%) 7{52} 2{5} 6{4} 90 22 mg 100 409.1450
409.1454 (54%) 7{53} 2{5} 6{5} 98 24 mg 100 435.1606 435.1616 (55%)
7{54} 2{5} 6{6} 79 44 mg 100 473.1399 473.1407 (92%) 7{55} 2{5}
6{7} 98 27 mg 99 444.1246 444.1255 (61%) 7{56} 2{5} 6{8} 90 11 mg
100 436.0653 436.0671 (24%) 7{57} 2{5} 6{9} 91 33 mg 100 429.1137
429.1152 (77%) 7{58} 2{5} 6{10} 84 35 mg 100 459.1242 459.1247
(75%) 7{59} 2{5} 6{11} 32 2 mg 98 497.0357 497.0356 (5%) 7{60} 2{5}
6{12} 77 44 mg 100 531.0621 531.0616 (82%) 7{61} 2{6} 6{1} 85 20 mg
100 415.2233 415.2246 (48%) 7{62} 2{6} 6{2} 66 33 mg 83 490.2706
490.2709 (67%) 7{63} 2{6} 6{3} 74 23 mg 100 449.2440 449.2453 (51%)
7{64} 2{6} 6{4} 76 13 mg 100 401.2440 401.2445 (34%) 7{65} 2{6}
6{5} 86 13 mg 100 427.2597 427.2602 (31%) 7{66} 2{6} 6{6} 66 23 mg
99 465.2389 465.2394 (49%) 7{67} 2{6} 6{7} 92 12 mg 99 436.2236
436.2261 (28%) 7{68} 2{6} 6{8} 69 16 mg 97 428.1644 428.1667 (37%)
7{69} 2{6} 6{9} 75 16 mg 100 421.2127 421.2134 (39%) 7{70} 2{6}
6{10} 70 27 mg 100 451.2233 451.2248 (64%) 7{71} 2{6} 6{11} 40 19
mg 97 489.1348 489.1344 (39%) 7{72} 2{6} 6{12} 58 32 mg 99 523.1611
523.1611 (61%) .sup.aInsoluble sample, purified by flash
chromatography
[0094] All chemicals were used as purchased from commercial
suppliers. Methylene chloride and THF were dried by being passed
through two packed columns of neutral alumina using the PurSolv
solvent purification system (Innovative Technology Inc.) prior to
use. (E)-3,5-hexadien-1-ol was prepared following the protocol of
Miller and Batey (Miller, C. A.; Batey, R. A. Org. Lett. 2004, 6,
699-702). (E)-3,5-hexadien-1-yl methanesulfonate and
N-benzyl-N-[(E)-3,5-hexadien-1-yl]amine 1(4) were prepared
following the protocol of Metz and coworkers (Plietker, B.; Seng,
D.; Frohlich, R; Metz, P. Tetrahedron 2000, 56, 873-879). The
chromatography solvent "wet ether" refers to the organic layer of a
9:1:0.1 ether:aqueous potassium phosphate, monobasic (0.5
M):glacial acetic acid mixture (Taber, D. F.; Pan, Y.; Zhao, X. J.
Org. Chem 2004, 69, 7234-7240).
[0095] The parallel syntheses were performed on the Bohdan
Miniblock XT parallel solution phase synthesizer obtained from
Mettler-Toledo Auto Chem. Automated weighing was performed using
the Bohdan Balance Automator (Mettler-Toledo Auto Chem). Parallel
evaporation was performed on the GeneVac EZ-2 plus evaporator
system. .sup.1H and .sup.13C NMR spectra were recorded on a Bruker
AM 400 spectrometer (operating at 400 and 100 MHz respectively) in
CDCl.sub.3 with 0.03% TMS as an internal standard. Chemical shifts
are reported in parts per million (ppm) downfield from TMS.
.sup.13C multiplicities were determined with the aid of an APT
pulse sequence, differentiating the signals for methyl and methane
carbons as "d" from methylene and quaternary carbons as "u". The
infrared (IR) spectra were acquired as thin on a PerkinElmer
Spectrum One FT-IR spectrometer and the absorption frequencies are
reported in cm.sup.-1. Melting points were determined on an
Electrothermal Mel-Temp model number 101D apparatus and are
uncorrected.
[0096] HPLC analysis was carried out using a Xterra MS C-18 column
(5 .mu.M, 4.6.times.150 mm) with gradient elution (10% CH.sub.3CN
to 100% CH.sub.3CN) on a Waters mass-directed fractionation
instrument using a Waters 2767 sample manager, a Waters 2525 HPLC
pump, a 2487 dual .lamda. absorbance detector and a
Waters/MicroMass ZQ (quadrupole) MS connected to a PC with a
MassLynx workstation. Purification was carried out using an Xterra
MS C-18 column (5 .mu.M, 19.times.150 mm) with narrow gradient
elution (acetonitrile and water) with a UV fraction trigger. High
resolution mass spectra (HRMS) [ESI+] were obtained using a
Waters/MicroMass ICT Premier (TOF instrument).
General Procedures for Preparing Aminodienes
##STR00017##
[0098] N-Butyl-N-[(E)-3,5-hexadien-1-yl]amine 1 (1).
(E)-3,5-hexadien-1-yl methanesulfonate (726.8 mg, 4.54 mmol) and
n-butylamine (4.5 mL, 40.54 mmol) were stirred in a sealed tube for
19 h at 65.degree. C. The reaction was partitioned between NaOH (1
N, 30 mL) and ether. The organics were dried (Na.sub.2SO.sub.4),
the solvent removed in vacuo and the residue purified by silica
chromatography to give 1(1) as a pale yellow oil (552.0 mg, 3.60
mmol, 79% yield). TLC R.sub.f=0.25 (CH.sub.2Cl.sub.2/acetone 1:1);
.sup.1H NMR .delta. 0.92 (t, J=7.3 Hz, 3H), 1.34 (m, 2H), 1.47 (m,
2H), 2.30 (m, 2H), 2.60 (t, J=7.0 Hz, 2H), 2.68 (t, J 7.6 Hz, 2H),
4.98 (d, J=10.0 Hz, 1H), 5.11 (d, J=16.9 Hz, 1H), 5.68 (m, 1H),
6.09-6.15 (m, 1H), 6.27-6.36 (m, 1H); .sup.13C NMR .delta. d 14.0,
132.6 (.times.2), 137.0; u 20.5, 32.3, 33.2, 49.2, 49.7, 115.4; IR
3253 (w), 3086 (m), 2958 (s), 2928 (s), 1652 (m) cm.sup.-1; HRMS
calcd for C.sub.10H.sub.20N 153.1596, obsd 154.1571.
##STR00018##
[0099] N-Cyclopropyl-N-[(E)-3,5-hexadien-1-yl]amine 1(2).
(E)-3,5-hexadien-1-yl methanesulfonate (515.7 mg, 3.22 mmol) and
cyclopropylamine (2.2 mL, 32.18 mmol) were stirred in a sealed tube
for 19 h at 65.degree. C. The reaction was partitioned between NaOH
(1 N, 30 mL) and ether. The organics were dried (Na.sub.2SO.sub.4),
the solvent removed in vacuo and the residue purified by silica
chromatography to give 1(2) as a pale yellow oil (175.4 mg, 1.28
mmol, 40% yield). TLC R.sub.f=0.40 (CH.sub.2Cl.sub.2/acetone 1:1);
.sup.1H NMR .delta. 0.29-0.33 (m, 2H), 0.39-0.43 (m, 2H), 2.07-2.12
(m, 1H), 2.24-2.30 (m, 2H), 2.75 (t, J=6.9 Hz, 2H), 4.97 (d, J=10.0
Hz, 1H), 5.09 (d, J=16.9 Hz, 1H), 5.62-5.69 (m, 1H), 6.07-6.13 (m,
1H), 6.25-6.34 (m, 1H); .sup.13C NMR .delta. d 30.2, 132.7, 132.7
(.times.2); u 6.4 (.times.2), 33.2, 48.9, 115.5; IR 3087 (m), 3008
(s), 2931 (s), 1652 (m), 1603 (m) cm.sup.-1; HRMS calcd for
C.sub.10H.sub.20N 138.1283, obsd 138.1244.
##STR00019##
[0100] N-Cyclohexyl-N-[(E)-3,5-hexadien-1-yl]amine 1(3).
(E)-3,5-hexadien-1-yl methanesulfonate (801.0 mg, 5.00 mmol) and
cyclohexylamine (5.7 mL, 50.00 mmol) were stirred in a sealed tube
for 19 h at 65.degree. C. The reaction was partitioned between NaOH
(1 N, 30 mL) and ether. The organics were dried (Na.sub.2SO.sub.4),
the solvent removed in vacuo and the residue purified by silica
chromatography to give 1(3) as a pale yellow oil (734.5 mg, 4.10
mmol, 82% yield). TLC R.sub.f=0.15 (CH.sub.2Cl.sub.2/acetone 1:1);
.sup.1H NMR .delta. 1.00-1.26 (m, 6H), 1.58-1.88 (m, 4H), 2.24-2.30
(m, 2H), 2.38-2.43 (m, 1H), 2.69 (t, J=7.0 Hz, 2H), 4.97 (d, J=10.0
Hz, 1H), 5.11 (d, J=16.4 Hz, 1H), 5.62-5.71 (m, 1H), 6.07-6.15 (m,
1H), 6.25-6.36 (m, 1H); .sup.13C NMR .delta. d 56.9, 132.7, 132.8,
137.2; u 25.2 (.times.2), 26.3, 33.6, 33.8 (.times.2), 46.3, 115.5;
IR 3085 (w), 2927 (s), 2853 (s), 1652 (m) cm.sup.-1; HRMS calcd for
C.sub.12H.sub.22N 180.1752, 180.1741 obsd
##STR00020##
[0101] N-(3,4-Dichlorobenzyl)-N-[(E)-3,5-hexadien-1-yl]amine 1(5).
(E)-3,5-hexadien-1-yl methanesulfonate (310.5 mg, 1.94 mmol) and
3,4-dichlorobenzylamine (1,023.4 mg, 5.81 mmol) and MeCN (1 mL)
were stirred in a sealed tube for 1 h at 130.degree. C. under
microwave irradiation. The reaction was partitioned between NaOH (1
N, 30 mL) and ether. The organics were dried (Na.sub.2SO.sub.4),
the solvent removed in vacuo and the residue purified by silica
chromatography to give 1(5) as a colorless oil (303.3 mg, 1.18
mmol, 61% yield). TLC R.sub.f=0.85 (CH.sub.2Cl.sub.2/acetone 1:1);
.sup.1H NMR .delta. 2.30 (m, 2H), 2.68 (t, J=6.5 Hz, 2H), 3.74 (s,
2H), 5.00 (d, J=8.6 Hz, 1H), 5.12 (d, J=18.4 Hz, 1H), 5.63-5.70 (m,
1H), 6.08-6.15 (m, 1H), 6.26-6.36 (m, 1H), 7.15 (dd, J=8.1, 2.0 Hz,
1H), 7.38 (d, J=8.2 Hz, 1H), 7.43 (d, J=1.7 Hz, 1H); .sup.13C NMR
.delta. d 127.4, 130.0, 130.3, 132.2, 132.9, 136.9; u 33.1, 48.5,
52.7, 115.7, 130.7, 132.4, 140.8; IR 3313 (w), 2914 (s), 2826 (s),
1806 (w), 1651 (m) cm.sup.-1; HRMS calcd for
C.sub.13H.sub.16Cl.sub.2N 256.0660, 256.0655 obsd.
##STR00021##
[0102] N-(3,4-Dimethoxybenzyl)-N-[(E)-3,5-hexadien-1-yl]amine 1(6).
(E)-3,5-hexadien-1-yl methanesulfonate (688.0 mg, 4.29 mmol) and
3,4-dichlorobenzylamine (2,513.0 mg, 15.03 mmol) and MeCN (2 mL)
were stirred in a sealed tube for 1 h at 130.degree. C. under
[0103] microwave irradiation. The reaction was partitioned between
NaOH (1 N, 30 mL) and ether. The organics were dried
(Na.sub.2SO.sub.4), the solvent removed in vacuo and the residue
purified by silica chromatography to give 1(6) as a light yellow
oil (686.1 mg, 2.77 mmol, 65% yield). TLC R.sub.f=0.34
(CH.sub.2Cl.sub.2/acetone 1:1); .sup.1H NMR .delta. 2.32 (m, 2H),
2.72 (t, J=7.1 Hz, 2H), 3.75 (s, 2H), 3.88 (s, 3H), 3.90 (s, 3H),
5.00 (d, J=10.1 Hz, 1H), 5.12 (d, J=17.0 Hz, 1H), 5.65-5.73 (m,
1H), 6.09-6.16 (m, 1H), 6.27-6.36 (m, 1H), 6.81-6.86 (m, 2H), 6.89
(d, J=1.44 Hz, 1H); .sup.13C NMR .delta. d 56.0, 56.1, 111.1,
111.5, 120.3, 132.7, 132.8, 137.1; u 33.2, 48.7, 53.8, 115.6,
133.2, 148.1, 149.1; IR 3318 (w), 3000 (s), 2934 (s), 2833 (s),
1801 (w), 1651 (m) cm.sup.-1; HRMS calcd for
C.sub.15H.sub.22NO.sub.2 248.1651, 248.1655 obsd.
General Procedures for the Tandem Diels-Alder/Acylation Reaction
Sequence
##STR00022##
[0105]
2-Butyl-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoquinoline-8-carboxylic
acid 2(1). Amino diene 1(1) (192.2 mg, 1.25 mmol) and maleic
anhydride (153.0 mg, 1.56 mmol) were dissolved in dichloroethane (3
mL) and heated at 165.degree. C. for 1.5 h under microwave
irradiation. The solvent was removed in vacuo and the residue
chromatographed to give 2(1) as a colorless oil (232.0 mg, 0.92
mmol, 74% yield). The product precipitated as a tan solid upon
trituration with hexanes. TLC R.sub.f=0.45 ("wet ether");
mp=96.5-101.5.degree. C.; .sup.1H NMR .delta. 0.93 (t, J=7.4 Hz,
3H), 1.29 (q, J=7.6 Hz, 2H), 1.48-1.53 (m, 2H), 1.90-2.04 (m, 2H),
2.28-2.46 (m, 2H), 2.82 (s, 1H), 2.89-2.90 (m, 1H), 3.11-3.33 (m,
4H), 3.49-3.57 (m, 1H), 5.57 (d, J=10.0 Hz, 1H), 5.88-5.93 (m, 1H);
.sup.13C NMR .delta. d 13.8, 34.6, 41.3, 44.7; 127.5, 129.3; u
19.9, 25.2, 27.0, 28.8, 44.4, 47.7, 171.1, 176.6; IR 2933 (m), 2253
(m), 1709 (s), 1625 (m) cm.sup.-1; HRMS calcd for
C.sub.14H.sub.22NO.sub.3 252.1600, 252.1606 obsd.
##STR00023##
[0106]
2-cyclopropyl-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoquinoline-8-carbo-
xylic acid 2(2). Amino diene 1(2) (165.0 mg, 1.20 mmol) and maleic
anhydride (147.5 mg, 1.50 mmol) were dissolved in dichloroethane (3
mL) and heated at 165.degree. C. for 1.5 h under microwave
irradiation. The solvent was removed in vacuo and the residue
chromatographed to give 2(2) as an off-white solid (215.9 mg, 0.92
mmol, 76% yield). TLC R.sub.f=0.35 ("wet ether");
mp=147-149.degree. C.; .sup.1H NMR .delta. 0.48-0.54 (m, 1H),
0.68-0.88 (m, 3H), 1.84-1.98 (m, 2H), 2.26-2.41 (m, 2H), 2.64-2.70
(m, 1H), 2.79-2.85 (m, 2H), 3.14-3.21 (m, 3H), 5.54 (d, J=8.6 Hz,
1H), 5.85-5.86 (m, 1H); .sup.13C NMR .delta. d 30.3, 34.3, 42.2,
43.6, 127.5, 129.4; u 6.4, 6.9, 24.6, 27.3, 44.4, 173.3, 177.6; IR
3053 (s), 2886 (s), 1708 (s), 1639 (m) cm.sup.-; HRMS calcd for
C.sub.13H.sub.18NO.sub.3 236.1287, 236.1293 obsd.
##STR00024##
[0107]
2-cyclohexyl-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoquinoline-8-carbox-
ylic acid 2(3). Amino diene 1(3) (399.0 mg, 2.23 mmol) and maleic
anhydride (272.8 mg, 2.78 mmol) were dissolved in dichloroethane (4
mL) and heated at 165.degree. C. for 1.5 h under microwave
irradiation. The solvent was removed in vacuo and the residue
chromatographed to give 2(3) as a very light yellow solid (423.6
mg, 1.53 mmol, 68% yield). TLC R.sub.f=0.56 ("wet ether");
mp=164.5-168.0.degree. C.; .sup.1H NMR .delta. 1.36-1.46 (m, 4H),
1.58-1.71 (m, 4H), 1.80-1.84 (m, 2H), 1.90-1.95 (m, 2H), 2.28-2.47
(m, 2H), 2.78-2.82 (m, 1H), 2.90-2.95 (m, 1H), 3.05-3.14 (m, 2H),
3.21-3.25 (m, 1H), 4.39-4.46 (m, 1H), 5.54 (d, J=10.1 Hz, 1H),
5.88-5.91 (m, 1H); .sup.13C NMR .delta. d 33.9, 41.8, 44.4, 53.6,
127.4, 129.2; u 24.9, 25.4, 25.5, 25.6, 27.0, 29.3, 29.5, 38.3,
170.6, 177.5; M 2930 (s), 2858 (s), 1.708 (s), 1619 (s) cm.sup.-1;
HRMS calcd for C.sub.16H.sub.24NO.sub.3 278.1756, 278.1785
obsd.
##STR00025##
[0108]
2-benzyl-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoquinoline-8-carboxylic
acid 2(4). Amino diene 1(4) (187.3 mg, 1.00 mmol) and maleic
anhydride (122.6 mg, 1.25 mmol) were dissolved in dichloroethane
(2.5 mL) and heated at 165.degree. C. for 1.5 h under microwave
irradiation. The solvent was removed in vacuo and the residue
chromatographed to give 2(4) as a very light yellow solid (210.9
mg, 0.74 mmol, 74% yield). TLC R.sub.f=0.56 ("wet ether");
mp=149.0-152.0.degree. C.; .sup.1H NMR .delta. 1.86-1.92 (m, 1H),
1.95-2.04 (m, 1H), 2.37-2.51 (m, 2H), 2.82-2.85 (m, 1H), 2.88-2.97
(m, 1H), 3.15-3.19 (m, 2H), 3.23-3.24 (m, 1H), 4.53 (d, J=14.6 Hz,
1H), 4.72 (d, J=14.9 Hz, 1H), 5.55 (dd, J=1.8, 10.0 Hz, 1H),
5.88-5.93 (m, 1H), 7.20 (d, J=7.9 Hz, 2H), 7.30-7.36 (m, 3H);
.sup.13C NMR .delta. d 34.8, 41.3, 45.4, 127.5, 127.7, 127.8,
128.7, 129.5; u 25.6, 27.0, 44.0, 51.0, 135.9, 172.0, 175.6; IR
3026 (s), 2923 (s), 1704 (s), 1635 (s) cm.sup.-1; HRMS calcd for
C.sub.17H.sub.20NO.sub.3 286.1443, 286.1452 obsd.
##STR00026##
[0109]
2-(3,4-dichlorobenzyl)-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoquinolin-
e-8-carboxylic acid 2(5). Amino diene 1(5) (256.2 mg, 1.00 mmol)
and maleic anhydride (122.6 mg, 1.25 mmol) were dissolved in
dichloroethane (2.5 mL) and heated at 165.degree. C. for 1.5 h
under microwave irradiation. The solvent was removed in vacuo and
the residue chromatographed to give 2(5) as a very light yellow
solid (283.4 mg, 0.80 mmol, 80% yield). TLC R.sub.f=0.63 ("wet
ether"); mp=208.0-209.5.degree. C.; NMR .delta. 1.89-1.95 (m, 1H),
2.00-2.09 (m, 1H), 2.35-2.51 (m, 2H), 2.86-2.91 (m, 2H), 3.12-3.22
(m, 2H), 3.29-3.31 (m, 1H), 4.31 (d, J=15.0 Hz, 1H), 4.79 (d,
J=15.0 Hz, 1H), 5.58 (d, J=10.0 Hz, 1H), 5.91-5.95 (m, 1H), 7.05
(dd, J=1.7, 8.2 Hz, 1H), 7.26 (d, J=2.1 Hz, 1H), 7.40 (d, J=8.2 Hz,
1H); .sup.13C NMR .delta. d 34.7, 41.6, 44.5, 127.1, 127.6, 129.5,
129.7, 130.7; u 25.2, 27.0, 44.4, 50.6, 131.8, 132.8, 136.4, 171.9,
175.7; IR 3434 (s), 2107 (m), 1699 (s), 1626 (s) cm.sup.-1; HRMS
calcd for C.sub.17H.sub.18NO.sub.3 354.0664, 354.0667 obsd.
##STR00027##
[0110]
2-(3,4-dimethoxybenzyl)-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoquinoli-
ne-8-carboxylic acid 2(6). Amino diene 1(6) (348.8 mg, 1.41 mmol)
and maleic anhydride (172.9 mg, 1.76 mmol) were dissolved in
dichloroethane (3.5 mL) and heated at 165.degree. C. for 1.5 h
under microwave irradiation. The solvent was removed in vacuo and
the residue chromatographed to give 2(6) as a fluffy white solid
(387.2 mg, 1.12 mmol, 80% yield). TLC R.sub.f=0.23 ("wet ether");
mp=142.5-144.0.degree. C.; .sup.1H NMR .delta. 1.86-1.91 (m, 1H),
1.96-2.04 (m, 1H), 2.40-2.48 (m, 2H), 2.82-2.86 (m, 1H), 2.91-2.96
(m, 1H), 3.13-3.17 (m, 2H), 3.21-3.24 (m, 1H), 3.87 (s, 3H), 3.89
(s, 3H), 4.35 (d, J=14.4 Hz, 1H), 4.76 (d, J=14.4 Hz, 1H), 5.54 (d,
J=10.3 Hz, 1H), 5.85-5.91 (m, 1H), 6.74-6.76 (m, 2H), 6.80-6.82 (m,
1H); .sup.13C NMR .delta. d 34.5, 41.8, 43.5, 55.8, 55.9, 110.9
(.times.2), 120.3, 127.8, 129.0; u 24.7, 26.9, 43.8, 50.6, 128.6,
148.5, 149.2, 171.1, 177.5; IR 2936 (s), 2253 (m), 1706 (s), 1627
(s) cm.sup.-1; HRMS calcd for C.sub.19H.sub.24NO.sub.5 346.1654,
346.1670 obsd.
##STR00028##
[0111]
2-Cyclopropyl-8-methyl-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoquinolin-
e-8-carboxylic acid 3(2). Amino diene 1(2) (98.2 mg, 0.716 mmol)
and citraconic anhydride (120.4 mg, 1.07 mmol) were dissolved in
dichloroethane (2.0 mL) and heated at 165.degree. C. for 1.5 h
under microwave irradiation. The solvent was removed in vacuo and
the residue chromatographed to give 3(2) as a sticky yellow gum
(91.4 mg, 0.367 mmol, 51% yield). TLC R.sub.f=0.57 ("wet ether");
mp=156.0-159.5.degree. C.; .sup.1H NMR .delta.0.40-0.45 (m, 1H),
0.67-0.72 (m, 2H), 0.77-0.83 (m, 1H), 1.23 (s, 3H), 1.89-2.00
(complex, 3H), 2.53-2.80 (m, 1H), 2.94 (d, J=5.6 Hz, 1H), 3.03-3.17
(m, 2H), 5.49 (d, J=9.6 Hz, 1H), 5.72-5.78 (m, 1H); .sup.13C NMR
.delta. d 23.1, 29.9, 30.3, 47.6, 126.0, 128.7; u 6.0, 7.1, 27.4,
30.1, 42.0, 44.5, 172.1, 181.7; IR 3448 (m), 3016 (s), 2935 (s),
1701 (s), 1627 (s) cm.sup.-1; HRMS calcd for
C.sub.14H.sub.20NO.sub.3 250.1443, 250.1452 obsd.
##STR00029##
[0112]
2-(3,4-Dichlorobenzyl)-8-methyl-1-oxo-1,2,3,4,4a,7,8,8a-octahydrois-
oquinoline-8-carboxylic acid 3(5)a and 3(5)b. Amino diene 1(5)
(200.0 mg, 0,781 mmol) and citraconic anhydride (175.0 mg, 1.56
mmol) were mixed together neat and heated at 100.degree. C. for 5.5
h in a conventional oil bath. The residue was dissolved in
CH.sub.2Cl.sub.2 and chromatographed to give 3(5)a and 3(5)b as a
sticky, tan-colored foam (219.8 mg, 0.597 mmol, 76% yield). TLC
R.sub.f=0.71 ("wet ether"); mp=186.0-189.0.degree. C.; .sup.1H NMR
.delta. 1.27 (s, 3H); 1.90-2.09 (m, 3H), 2.55-2.64 (m, 1H),
2.79-2.85 (m, 1H), 3.04-3.08 (m, 3H), 4.13 (d, J=15.2 Hz, 1H), 4.84
(d, J=15.2 Hz, 1H), 5.53 (d, J=9.8 Hz, 1H), 5.80-5.84 (m, 1H), 6.99
(dd, J=2.0, 8.1 Hz, 2H), 7.20 (d, J=2.0 Hz, 1H), 7.34 (d, J=8.4 Hz,
1H); .sup.13C NMR .delta. d 23.1, 30.6, 47.3, 126.2, 127.1, 128.9,
129.4, 130.5; u 26.9, 30.2, 42.2, 44.4, 49.6, 131.2, 135.0, 137.3,
170.3, 181.7; IR 3054 (m), 2918 (m), 2254 (m), 1698 (s), 1629 (s)
cm.sup.-1; HRMS calcd for C.sub.18H.sub.20Cl.sub.2NO.sub.3
368.0820, 368.0824 obsd.
##STR00030##
[0113] Methyl
2-(3,4-dichlorobenzyl)-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoquinoline-8-ca-
rboxylate 4a and 4b. Amino diene 1(5) (200.0 mg, 0.781 mmol) and
dimethyl fumarate (150.0 mg, 1.04 mmol) were dissolved in
dichloroethane (3.5 mL) and heated at 165.degree. C. for 1.5 h
under microwave irradiation. The solvent was removed in vacuo and
the residue chromatographed to give the diastereomeric mixture 4a
and 4b as a colorless oil (261.7 mg, 0.711 mmol, 91% yield). TLC
R.sub.f=0.84 (5% MeOH in CHCl.sub.3); .sup.1H NMR .delta. 1.69 (m,
1H), 1.83 (m, 1H), 2.00 (m, 1H), 2.20 (m, 1H), 2.30-2.33 (m, 1H),
2.42 (m 1H), 2.54-2.61 (m, 1H), 3.14-3.19 (m, 1H), 3.20-3.24 (m,
1H), 3.30 (m, 11-1), 3.58 (m, 1H), 3.72 (s, 3H), 3.80 (m, 1H);
.sup.13C NMR .delta. 23.5, 27.0, 28.9, 29.8, 30.0, 34.6, 39.4,
40.4, 41.7, 44.6, 45.5, 45.6, 48.8, 49.8, 51.9, 126.0, 126.9,
127.4, 127.5, 128.5, 128.7, 129.3, 129.8, 130.4, 130.6, 131.2,
131.4, 132.5, 132.6, 137.4, 137.4, 170.0, 170.9, 174.7, 176.8; IR
3022, 2945, 2934, 2845, 1734, 1639 cm.sup.-1; HRMS calcd for
C.sub.18H.sub.20Cl.sub.2NO.sub.3 368.0820, 368.0825 obsd.
##STR00031##
[0114] Methyl
2-(3,4-dichlorobenzyl)-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoquinoline-8-ca-
rboxylate 5. To the carboxylic acid scaffold 2(5) (67.1 mg, 0.189
mmol) in MeOH (0.5 mL) and benzene (1.5 mL) was added
trimethylsilyldiazomethane solution (0.47 mL, 2 M in ether, 0.945
mmol) and the reaction stirred for 14 h at rt. The solvent was
removed in vacuo and the residue chromatographed to 5 as a
colorless oil (68.8 mg, 0.187 mmol, 98% yield). TLC R.sub.f=0.09
(25% EtOAc in Hexanes); .sup.1H NMR .delta. 1.82-1.89 (m, 1H),
1.99-2.08 (m, 1H), 2.36-2.39 (m, 1H), 2.62-2.67 (m, 1H), 2.86 (m,
1H), 3.01-3.13 (m, 2H), 3.42-3.44 (m, 1H), 3.77 (s, 3H), 4.15 (d,
J=15.2 Hz, 1H), 4.83 (d, J=15.2 Hz, 1H), 5.55 (dd, J=0.8, 10.1 Hz,
1H), 5.87-5.91 (m, 1H), 6.99 (dd, J=1.6, 7.9 Hz, 1H), 7.21 (d,
J=1.2 Hz, 1H), 7.34 (d, J=8.3 Hz, 1H); .sup.13C NMR .delta. d 34.2,
41.0, 42.7, 51.8, 126.9, 127.6, 129.2, 129.3, 130.4; u 23.6, 27.2,
44.1, 49.3, 131.0, 132.4, 137.2, 169.6, 174.1; IR 3509 (w), 3021
(m), 2928 (m), 2869 (m), 2250 (m), 1736 (s), 1634 (s) cm.sup.-1;
HRMS calcd for C.sub.18H.sub.20Cl.sub.2NO.sub.3 368.0820, 368.0825
obsd.
[0115] General procedure for the coupling of carboxylic acid
scaffolds 2(1-6) with amines 6(1-12). Each reaction tube of a
24-position Bohdan Miniblock XT was charged with EDC.HCl (28.8 mg,
0.15 mmol). The air atmosphere was exchanged for argon and
CH.sub.2Cl.sub.2 (0.3 mL) was added. A solution of the carboxylic
acid scaffold 2(1-61 (0.10 mmol) in CH.sub.2Cl.sub.2 (0.4 mL) was
added via syringe followed by a solution of the amine 6(1-12) (0.20
mmol) and DMAP (mg, 0.02 mmol) in CH.sub.2Cl.sub.2 (0.4 mL). The
reactions were stirred at room temperature (rt) for 14 h then
partitioned between CH.sub.2Cl.sub.2 and water in hydrophobic phase
separator tubes, which allowed the halogenated solvent layer to
pass through. The aqueous layers were washed with CH.sub.2Cl.sub.2
(2.times.3 mL). The combined organics were passed through 1,000 mg
silica SPE tubes, eluting with CH.sub.2Cl.sub.2 (7 mL) then
CH.sub.2Cl.sub.2:acetone (1:1, 7 mL) to yield the crude amides
7(1-72). The crude amides were evaporated in a Genevac EZ-2 Plus
parallel evaporator and subjected to mass-directed preparative HPLC
purification to give the pure amides 7(1-72). Characterization of
representative library examples:
##STR00032##
[0116]
2-Butyl-8-(morpholine-4-carbonyl)-2,3,4,4a,8,8a-hexahydroisoquinoli-
n-1(7H)-one 7(1). .sup.1H NMR .delta. 0.89 (t, J=8.0 Hz, 3H), 1.21
(q, J=8.0 Hz, 2H), 1.42-1.50 (m, 2H), 1.84-2.01 (m, 2H), 2.14-2.20
(m, 1H), 2.40-2.49 (m, 1H), 2.78-2.80 (m, 1H), 2.89-2.97 (m, 2H),
3.06-3.13 (m, 2H), 3.27-3.34 (m, 1H), 3.47-3.53 (m, 2H), 3.55-3.60
(m, 1H), 3.65-3.71 (m, 6H), 5.53 (dd, 0.1=2.3, 8.0 Hz, 1H),
5.89-5.94 (m, 1H); .sup.13C NMR .delta. d 13.9, 34.6, 38.5, 42.6,
127.5, 128.9; u 20.0, 25.0, 27.3, 29.1 (.times.2), 44.6, 47.3
(.times.2), 66.9 (.times.2), 168.2, 172.3.
##STR00033##
[0117]
2-Cyclopropyl-8-(morpholine-4-carbonyl)-2,3,4,4a,8,8a-hexahydroisoq-
uinolin-1 (7H)-one 7(13). .sup.1H NMR .delta. 0.55-0.59 (m, 2H),
0.70-0.81 (m, 2H), 1.80-1.88 (m, 1H), 1.99-2.06 (m, 1H), 2.14-2.21
(m, 1H), 2.35-2.44 (m, 1H), 2.69-2.75 (m, 2H), 2.86-2.89 (ma, 1H),
3.00-3.04 (m, 1H), 3.09-3.15 (m, 1H), 3.19-3.25 (m, 1H), 3.50-3.56
(m, 2H), 3.65-3.70 (m, 6H), 5.51 (dd, J=2.2, 10.1 Hz, 1H),
5.85-5.90 (m, 1H); .sup.13C NMR .delta. d 30.1, 34.1, 38.3, 42.9,
127.7, 128.3; u 6.0, 6.6, 25.1 (.times.2), 27.3, 44.8 (.times.2),
66.9 (.times.2), 170.7, 172.2.
##STR00034##
[0118]
2-Cyclohexyl-8-(morpholine-4-carbonyl)-2,3,4,4a,8,8a-hexahydroisoqu-
inolin-1(7H)-one 7(25). .sup.1H NMR .delta. 0.99-1.10 (m, 1H),
1.30-1.38 (m, 4H), 1.53-1.55 (n, 1H), 1.61-1.63 (m, 2H), 1.73-1.86
(m, 3H), 1.91-1.97 (n, 1H), 2.12-2.20 (m, 1H), 2.41-2.50 (m, 1H),
2.75-2.81 (m, 1H), 2.89-2.95 (m, 2H), 3.10-3.13 (m, 2H), 3.47-3.51
(m, 2H), 3.67-3.77 (m, 6H), 4.45-4.53 (m, 1H), 5.49 (dd, J=2.2,
10.1 Hz, 1H), 5.90-5.95 (n, 1H); .sup.13C NMR .delta. d 34.1, 39.0,
43.0, 52.2, 127.4, 129.2; u 24.8, 25.6 (.times.2), 25.7, 27.5,
29.3, 30.0 (.times.2), 38.1 (.times.2), 66.9 (.times.2), 167.8,
172.4.
##STR00035##
[0119]
2-Benzyl-8-(morpholine-4-carbonyl)-2,3,4,4a,8,8a-hexahydroisoquinol-
in-1(7H)-one 7(37). .sup.1H NMR .delta. 1.84-1.92 (n, 1H),
1.94-2.00 (m, 1H), 2.21-2.27 (m, 1H), 2.50-2.59 (m, 1H), 2.77-2.83
(m, 1H), 2.98-3.07 (m, 3H), 3.12-3.19 (m, 1H), 3.51-3.54 (m, 2H),
3.66-3.74 (m, 6H), 4.35 (d, J=14.6 Hz, 1H), 4.83 (d, J=14.6 Hz,
1H), 5.50 (dd, J=1.9, 10.1 Hz, 1H), 5.88-5.93 (m, 1H), 7.18-7.31
(m, 5H); .sup.13C NMR .delta. d 34.6, 38.5, 42.6, 127.2, 127.7,
127.9 (.times.2), 128.5 (.times.3); u 25.2 (.times.2), 27.2, 44.2,
50.4 (.times.2), 66.9 (.times.2), 137.1, 168.8, 172.3.
##STR00036##
[0120]
2-(3,4-Dichlorobenzyl-8-(morpholine-4-carbonyl)-2,3,4,4a,8,8a-hexah-
ydroisoquinolin-1(7H)-one 7(49). .sup.1H NMR .delta. 1.87-1.95 (m,
1H), 2.05-2.12 (m, 1H), 2.25-2.31 (n, 1H), 2.48-2.57 (m, 1H),
2.82-2.84 (m, 1H), 2.97-3.00 (n, 1H), 3.07-3.13 (m, 2H), 3.16-3.22
(m, 1H), 3.51-3.52 (m, 2H), 3.67-3.70 (m, 6H), 4.18 (d, J=14.9 Hz,
1H), 4.89 (d, J=14.9 Hz, 1H), 5.55 (dd, J=2.0, 10.0 Hz, 1H),
5.90-5.95 (m, 1H), 7.05 (dd, J=2.1, 10.2 Hz, 1H), 7.30 (d, J=2.0
Hz, 1H), 7.37 (d, J=8.5 Hz, 1H); .sup.13C NMR .delta. d 34.5, 38.4,
42.4, 127.1, 127.9, 128.2, 129.5, 130.5; u 25.2 (.times.2), 27.2,
44.8, 49.5 (.times.2), 66.9 (.times.2), 131.2, 132.6, 137.5, 169.3,
172.1.
##STR00037##
[0121]
2-(3,4-Dimethoxybenzyl-8-(morpholine-4-carbonyl)-2,3,4,4a,8,8a-hexa-
hydroisoquinolin-1(7H)-one 7(61). .sup.1H NMR .delta. 1.84-1.92 (m,
1H), 1.95-2.02 (m, 1H), 2.21-2.28 (m, 1H), 2.50-2.59 (m, 1H),
2.80-2.81 (in, 1H), 2.97-3.06 (m, 3H), 3.09-3.16 (m, 1H), 3.52-3.54
(m, 2H), 3.68-3.73 (m, 6H), 3.86 (s, 3H), 3.87 (s, 3H), 4.08 (d,
J=14.3 Hz, 1H), 4.99 (d, J=14.3 Hz, 1H), 5.50 (dd, J=2.0, 9.9 Hz,
1H), 5.85-5.90 (m, 1H), 6.72-6.79 (m, 3H); .sup.13C NMR .delta. d
34.6, 38.6, 42.6, 55.9 (.times.2), 110.8, 111.0, 120.3, 128.0,
128.3; u 25.1 (.times.2), 27.2, 44.0, 50.1 (.times.2), 66.9
(.times.2), 129.7, 148.3, 149.2, 168.8, 172.3.
##STR00038##
[0122]
2-Benzyl-8-(4-phenylpiperazine-1-carbonyl)-2,3,4,4a,8,8a-hexahydroi-
soquinolin-1(7H)-one 7(38). .sup.1H NMR .delta. 1.64-1.76 (m, 1H),
1.86-1.95 (m, 1H), 1.98-2.05 (m, 1H), 2.26-2.30 (m, 1H), 2.54-2.62
(in, 1H), 2.85-2.86 (m, 1H), 3.05-3.28 (8H), 3.68-3.74 (m, 2H),
3.81-3.96 (m, 1H), 4.37 (d, J 14.6 Hz, 1H), 4.85 (d, J=14.6 Hz,
1H), 5.53 (dd, J=1.8, 9.9 Hz, 1H), 5.91-5.96 (m, 1H), 6.90-6.96 (m,
3H), 7.20-7.32 (m, 7H); .sup.13C NMR .delta. d 34.6, 38.6, 42.8,
116.5 (.times.2), 120.3, 127.2, 127.7, 127.9 (.times.2), 128.4
(.times.2), 128.6, 129.2 (.times.2); u 25.2 (.times.2), 27.2, 44.2
(.times.2), 49.6, 50.3 (.times.2), 137.1, 151.1, 168.8, 172.1.
##STR00039##
[0123]
2-Benzyl-8-(phenethyl-4-carbonyl)-2,3,4,4a,8,8a-hexahydroisoquinoli-
n-1(7H)-one 7(39). .sup.1H NMR .delta. 1.75-1.81 (m, 1H), 1.92-2.00
(m, 2H), 2.26-2.30 (m, 1H), 2.65-2.70 (m, 1H), 2.80-2.88 (m, 3H),
3.08 (dd, J=3.7, 8.7 Hz, 2H), 3.52-3.58 (m, 2H), 4.43 (d, J=14.6
Hz, 1H), 5.55 (dd, J=1.7, 9.9 Hz, 1H), 5.81-5.85 (m, 1H), 7.15-7.30
(m, 10H), 7.37 (br s, 1H); .sup.13C NMR .delta. d 35.6, 42.4, 44.9,
426.2, 127.3, 127.6 (.times.2), 128.4 (.times.3), 128.5 (.times.2),
128.7, 128.9 (.times.2); u 25.3, 27.3, 35.6, 40.9, 44.2, 50.3,
136.9, 139.5, 170.5, 174.0.
##STR00040##
[0124]
2-Benzyl-N-butyl-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoquinoline-8-ca-
rboxamide 7(40). .sup.1H NMR .delta. 0.92 (t, J=7.3 Hz, 3H),
1.34-1.41 (m, 2H), 1.48-1.55 (m, 2H), 1.77-1.87 (m, 1H), 1.92-2.03
(m, 1H), 2.34-2.40 (m, 2H), 2.71-2.75 (m, 1H), 2.78-2.86 (m, 1H),
3.08-3.11 (m, 2H), 3.23-336 (m, 3H), 4.46 (d, J=14.9 Hz, 1H), 4.69
(d, J=14.9 Hz, 1H), 5.56 (dd, J=1.8, 10.1 Hz, 1H), 5.85-5.88 (m,
1H), 7.16-7.18 (m, 2H), 7.25-7.32 (m, 3H), 7.55 (br s, 1H);
.sup.13C NMR .delta. d 13.8, 35.7, 42.5, 45.3, 127.3, 127.7
(.times.2), 128.4, 128.5 (.times.2), 128.9; u 20.2, 25.7, 27.2,
31.6, 39.4, 44.3, 50.4, 136.8, 170.8, 174.1.
##STR00041##
[0125]
2-Benzyl-N-cyclohexyl-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoquinoline-
-8-carboxamide 7(41). .sup.1H NMR .delta. 1:12-1.26 (m, 3H),
1.32-1.42 (m, 2H), 1.57-1.62 (m, 1H), 1.66-1.82 (m, 3H), 1.90-1.99
(m, 4H), 2.33-2.36 (m, 2H), 2.70-2.84 (m, 2H), 3.06-3.10 (m, 2 H),
3.20-3.22 (m, 1H), 3.74-3.83 (m, 1H), 4.50 (d; J=14.7 Hz, 1H), 4.65
(d, J=14.6 Hz, 1H), 5.55 (dd, J=1.8, 9.9 Hz, 1H), 5.83-5.88 (m,
1H), 7.16-7.18 (m, 2H), 7.24-7.32 (m, 3H), 7.43 (br s, 1H);
.sup.13C NMR .delta. d 35.7, 42.5, 45.2, 48.1, 127.3, 127.8
(.times.2), 128.3, 128.5 (.times.2), 128.8; u 24.9 (.times.2),
25.7, 25.8, 27.2, 32.9 (.times.2), 44.4, 50.4, 136.9, 170.8,
173.1.
##STR00042##
[0126]
2-Benzyl-N-(4-methoxybenzyl)-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoqu-
inoline-8-carboxamide 7142). .sup.1H NMR .delta. 1.75-1.81 (m, 1H),
1.91-2.01 (m, 1H), 2.35-2.37 (m, 2H), 2.73-2.80 (m, 2H), 3.04-3.08
(m, 2H), 3.28-3.29 (m, 1H), 3.79 (s, 3H), 4.35-4.40 (m, 2H),
4.45-4.53 (m, 2H), 4.59 (d, J=14.7 Hz, 1H), 5.56 (dd, J=2.0, 9.9
Hz, 1H), 5.84-5.87 (m, 1H), 6.85 (d, J=8.6 Hz, 2H), 7.15 (d, J=8.4
Hz, 2H), 7.24-7.31 (m, 5H), 7.61 (br s, 1H); .sup.13C NMR .delta. d
35.5, 42.5, 44.7, 55.3, 113.9 (.times.2), 127.3, 127.7 (.times.2),
128.4, 1'28.5 (.times.2), 128.6, 129.1 (.times.2); u 25.4, 27.2,
43.0, 44.3, 50.4, 131.1, 136.9, 158.7, 170.6, 174.0.
##STR00043##
[0127]
2-Benzyl-N-(pyridin-3-ylmethyl)-1-oxo-1,2,3,4,4a,7,8,8a-octahydrois-
oquinoline-8-carboxamide 7(43). .sup.1H NMR .delta. 1.76-1.83 (m,
1H), 1.93-2.01 (m, 1H), 2.36-2.41 (m, 2H), 2.77-2.82 (m, 2H),
3.08-3.11 (m, 2H), 3.25-3.27 (m, 1H), 4.45 (d, J=14.9 Hz, 1H),
4.50-4.53 (m, 2H), 4.65 (d, J=14.9 Hz, 1H), 5.57 (dd, J=1.7, 10.0
Hz, 1H), 5.84-5.98 (m, 1H), 7.16 (d, J=6.8 Hz, 2H), 7.23-7.32 (m,
4H), 7.73 (d, J=7.8 Hz, 1H), 8.22 (br s, 1H), 8.49 (d, J=6.3 Hz,
1H), 8.57 (d, J=2.0 Hz, 1H), .sup.13C NMR .delta. d 35.7, 42.5,
45.3, 123.5, 127.3, 127.6 (.times.2), 128.3, 128.6 (.times.2),
128.8, 135.5, 148.5, 149.1; u 25.7, 27.2, 41.0, 44.3, 50.5, 134.6,
136.7, 170.7, 174.5.
##STR00044##
[0128]
2-Benzyl-N-(thiazol-2-yl)-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoquino-
line-8-carboxamide 7(441. .sup.1H NMR .delta. 1.81-1.87 (m, 1H),
1.94-2.04 (in, 1H), 2.49-2.53 (in, 2H), 2.84-2.902 (m, 1H),
2.96-3.00 (m, 1H), 3.09-3.12 (m, 2H), 3.30-3.31 (m, 1H), 4.41 (d,
J=14.7 Hz, 1H), 4.80 (d, J=14.6 Hz, 1H), 5.58 (d, J=10.1 Hz, 1H),
5.88-5.93 (m, 1H), 6.93 (d, J=3.5 Hz, 1H), 7.17 (d, J=8.1 Hz, 2H),
7.23-7.31 (m, 3H), 7.46 (d, J=7.8 Hz, 1H), 12.55 (br s, 1H);
.sup.13C NMR .delta. d 35.6, 42.3, 46.0, 113.2, 127.4, 127.8
(.times.2), 128.0, 128.5 (.times.2), 128.9, 137.5; u 25.6, 27.2,
43.9, 50.6, 136.5, 159.0, 170.4, 172.0.
##STR00045##
[0129]
2-Cyclopropyl-N-(thiazol-2-yl)-1-oxo-1,2,3,4,4a,7,8,8a-octahydroiso-
quinoline-8-carboxamide 7(20). .sup.1H NMR .delta. 0.46-0.52 (m,
1H), 0.66-0.84 (m, 3H), 1.83-1.88 (m, 1H), 1.90-1.99 (m, 1H),
2.42-2.46 (m, 2H), 2.64-2.70 (m, 1H), 2.79-2.87 (m, 1H), 2.90-2.94
(m, 1H), 3.08-3.21 (in, 3H), 5.58 (dd, J=1.8, 10.1 Hz, 1H),
5.86-5.91 (m, 1H), 6.93 (d, J=3.5 Hz, 1H), 7.45 (d, 0.1=3.5 Hz,
1H), 12.15 (br s, 1H); .sup.13C NMR .delta. d 30.2, 35.2, 42.6,
45.7, 113.2, 127.9, 128.8, 137.5; u 6.4, 6.8, 25.3, 27.4, 44.4,
158.8, 172.0, 172.4.
##STR00046##
[0130]
2-Cyclohexyl-N-(thiazol-2-yl)-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoq-
uinoline-8-carboxamide 7(32), .sup.1H NMR .delta. 1.00-1.09 (m,
1H), 1.30-1.40 (m, 4H), 1.54-1.66 (m, 3H), 1.74-1.78 (m, 2H),
1.84-2.02 (m, 2H), 2.41-2.46 (m, 2H), 2.78-2.87 (m, 1H), 2.92-2.97
(m, 1H), 3.01-3.08 (m, 1H), 3.15-3.21 (m, 2H), 4.49-4.54 (m, 1H),
5.56 (d, J=9.8 Hz, 1H), 5.87-5.92 (m, 1H), 6.92 (d, J=3.6 Hz, 1H),
7.44 (d, J=3.5 Hz, 1H), 12.70 (br s, 1H); .sup.13C NMR .delta. d
34.9, 42.4, 46.0, 52.9, 113.1, 127.8, 128.8, 137.5; u 25.4, 25.6
(.times.2), 25.7, 27.4, 29.5, 29.6, 38.2, 159.0, 169.5, 172.2.
##STR00047##
[0131]
2-Benzyl-N-phenyl-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoquinoline-8-c-
arboxamide 7(45). .sup.1H NMR .delta. 1.81-1.85 (m, 1H), 1.94-2.02
(m, 1H), 2.46-2.55 (m, 2H), 2.83-2.85 (m, 1H), 2.93-2.97 (m, 1H),
3.12-3.14 (m, 2H), 3.22-3.26 (m, 1H), 4.48 (d, J=14.9 Hz, 1H), 4.78
(d, J=14.6 Hz, 1H), 5.58 (d, J=9.8 Hz, 1H), 5.90-5.93 (m, 1H), 7.06
(d, J=3.5 Hz, 1H), 7.17 (d, J=7.1 Hz, 2H), 7.26-7.32 (m, 5H), 7.636
(d, J=9.4 Hz, 2H), 10.89 (br s, 1H); .sup.13C NMR .delta. d 36.1,
42.5 (.times.2), 120.0 (.times.2), 123.6, 127.4, 127.7 (.times.2),
128.1, 128.6 (.times.2), 128.8 (.times.2), 129.3; u 26.6, 27.2,
44.4, 50.7, 136.6, 138.9, 171.4, 172.5.
##STR00048##
[0132]
2-Benzyl-N-(4-methoxyphenyl)-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoqu-
inoline-8-carboxamide 7(46). .sup.1H NMR .delta. 1.80-1.85 (m, 1H),
1.93-2.01 (m, 1H), 2.46-2.50 (m, 2H), 2.81-2.85 (m, 1H), 2.90-2.95
(m, 1H), 3.11-3.14 (m, 2H), 3.22-3.24 (m, 1H), 3.78 (s, 3H), 4.47
(d, J=14.9 Hz, 1H), 4.77 (d, J=14.9 Hz, 1H), 5.57 (dd, J=1.9, 10.0
Hz, 1H), 5.88-5.93 (n, 1H), 6.85 (d, J=9.1 Hz, 2H), 7.17 (d, J=8.1
Hz, 2H), 7.25-7.29 (m, 3H), 7.54 (d, J=9.1 Hz, 2H), 10.63 (br s,
1H); .sup.13C NMR .delta. d 36.1, 42.5 (.times.2), 55.5, 114.0
(.times.2), 121.6 (.times.2), 127.4, 127.7 (.times.2), 128.1, 128.6
(.times.2), 129.3; u 26.5, 27.2, 44.3, 50.6, 132.2, 136.6, 155.9,
171.4, 172.2.
##STR00049##
[0133]
2-Benzyl-N-(2,4-dichlorophenyl)-1-oxo-1,2,3,4,4a,7,8,8a-octahydrois-
oquinoline-8-carboxamide 7(47). .sup.1H NMR .delta. 1.82-1.91 (m,
1H), 2.00-2.05 (m, 1H), 2.46-2.58 (m, 2H), 2.89-2.95 (m, 2H),
3.10-3.21 (m, 2H), 3.38-3.48 (m, 1H), 4.48 (d, J=14.4 Hz, 1H), 4.68
(d, J=14.7 Hz, 1H), 5.62 (d, J=10.4 Hz, 1H), 5.89-5.94 (m, 1H),
7.15-7.36 (complex, 7H), 8.43 (d, J=9.1 Hz, 2H), 9.47 (br s, 1H);
.sup.13C NMR .delta. d 35.4, 42.6, 45.8, 122.9, 127.4, 127.7, 127.8
(.times.2), 128.3, 128.5, 128.6 (.times.2), 128.7; u 25.3, 27.3,
44.2, 50.4, 124.1, 128.7, 134.2, 136.8, 170.2, 172.5.
##STR00050##
[0134]
2-Benzyl-N-(2,4-dichlorophenyl)-1-oxo-1,2,3,4,4a,7,8,8a-octahydrois-
oquinoline-8-carboxamide 7(48). .sup.1H NMR .delta. 1.82-1.88 (m,
1H), 1.94-2.04 (m, 1H), 2.46-2.50 (m, 2H), 2.84-2.90 (m, 1H),
2.91-2.96 (m, 1H), 3.13-3.17 (m, 2H), 3.20-3.21 (m, 1H), 4.45 (d,
J=14.9 Hz, 1H), 4.82 (d, J=14.6 Hz, 1H), 5.58 (dd, J=1.8, 10.1 Hz,
1H), 5.89-5.94 (m, 1H), 7.17-7.19 (m, 2H), 7.26-7.33 (m, 3H), 7.40
(d, J=8.6 Hz, 1H), 7.80 (dd, J=2.8, 8.8 Hz, 1H), 7.99 (d, J=2.5 Hz,
1H), 11.73 (br s, 1H); .sup.13C NMR .delta. d 36.1, 42.2, 48.5,
118.8, 118.9, 123.8, 127.5 (.times.2), 128.0, 128.6 (.times.2),
129.3, 131.6; u 26.6, 27.1, 44.2, 50.8, 121.3, 124.1, 136.2, 137.9,
171.4, 172.9.
Near-Neat Procedure Diels-Alder/Acylation Reaction Sequence
##STR00051##
[0136] Reaction Scheme 5 shows a reaction where a diene 1(4) (4.9
g, 26 mmol) was dissolved in 5 mL dichloromethane in a 100 mL round
bottom flask fitted with a magnetic stir bar. Maleic anhydride 2
(2.8 g, 29 mmol) was added to the reaction flask in 5 portions over
5 minutes with constant stirring (the reaction is exothermic and
becomes violent if maleic anhydride is added all at once). After
stirring for additional 5 minutes, 60 mL hexane was added to the
reaction and the organic layer was decanted. The white solid
residue was dissolved in 50 mL 20% dichloromethane/hexane solution
which upon slow evaporation afforded 2(4) as colorless crystals
(6.6 g, 23 mmol, 88% yield).
Synthesis of Analogs from N-Substituted Maleimides
##STR00052##
[0137] Reaction Scheme 6 shows a reaction where diene 1(4) (100 mg,
0.53 mmol) and N-phenylmaleimide (104 mg, 0.60 mmol) were dissolved
in toluene (5 mL) in a 25 mL round bottom flask at room
temperature. The reaction was stirred for 5 hours at 80.degree. C.
The reaction was cooled and then directly loaded onto a silica gel
column. Chromatography with 20% EtOAc/hexane afforded 7(45) (164
mg, 0.45 mmol, 85% yield) as a white solid. The following compounds
were also synthesized using Scheme 6.
##STR00053##
One Pot Three Component Synthesis of Analogs
##STR00054##
[0139] Reaction Scheme 7 shows another diene, (E)-hexa-3,5-dienyl
methanesulfonate (100 mg, 0.57 mmol) and N-phenylmaleimide (118 mg,
0.68 mmol) were dissolved in toluene (5 mL) in a 25 mL round bottom
flask at room temperature. After stirring for 3 hours at 80.degree.
C., benzyl amine (0.18 mL, 177 mg, 1.65 mmol) was added and the
reaction was stirred for additional 6 h at 80.degree. C. The
reaction was cooled and then directly loaded onto silica gel
column. Chromatography with 20% EtOAc/hexane produced the
corresponding analog 7(45) (160 mg, 0.44 mmol, 78% yield) as a
white solid. The following compounds were also synthesized using
Scheme 7.
##STR00055##
Synthesis of the Lactone Carboxylic Acid Scaffold
##STR00056##
[0141] Reaction Scheme 8 produces a
1-oxo-3,4,4a,7,8,8a-hexahydro-1H-isochromene-8-carboxylic acid 8.
Maleic anhydride (2.30 g, 0.023 mol) was added to neat
(E)-hexa-3,5-dien-1-ol (2.30 g, 0.023 mol) in a glass mortar and
ground together with a pestal for 90 seconds to initiate the
reaction. After the initially vigorous exothermic reaction was
complete, the reaction was allowed to stand at rt for 30 minutes.
The crude reaction mixture was chromatographed to yield the lactone
carboxylic acid 8 as a white solid (3.42 g, 0.017 mol, 74%
yield).
Coupling of Carboxylic Acid Scaffolds with Alcohols
##STR00057##
[0142] Reaction Scheme 9 produces a
4-chloro-3-(trifluoromethyl)phenyl
2-benzyl-1-oxo-1,2,3,4,4a,7,8,8a-octahydroisoquinoline-8-carboxylate
9(1). A 20 mL scintillation vial was charged with a mixture of the
carboxylic acid (145 mg, 0.51 mmol), the phenol (200 mg, 1.02
mmol), EDC.HCl (244 mg, 1.28 mmol) and DMAP (12 mg, 0.10 mmol). A
solution of Et.sub.3N (205 mg, 1.02 mmol) in CH.sub.2Cl.sub.2 (10
mL) was added, the vial was capped and stirred at rt for 14 h. The
reaction was partitioned between water (25 mL) and CH.sub.2Cl.sub.2
(3.times.10 mL) and the combined organic layers were dried with
Na.sub.2SO.sub.4. The crude product was purified by silica gel
chromatography to afford the ester derivative 9(1) as a colorless
oil (174 mg, 0.38 mmol, 74% yield).
##STR00058## ##STR00059##
[0143] Scheme 10 illustrates the resolution of racemic carboxylic
acid scaffold 2(4) to its enantiomers 2(4a) and 2(4b). To a
solution of carboxylic acid 2(4) (285 mg, 1.0 mmol) in dry
CH.sub.2Cl.sub.2 (20 mL) were added EDC (382 mg, 2.0 mmol), DMAP
(12 mg, 0.1 mmol) and (R)-1-Phenyl-2-propyn-1-ol (160 mg, 1.2 mmol)
at room temperature. After stirring for 16 hour at room temperature
the reaction solution was diluted with CH.sub.2Cl.sub.2 (80 mL),
washed with water (20 mL.times.2) and the combined organic layers
were dried over anhydrous MgSO.sub.4. Evaporation of solvent
followed by chromatography with 10% EtOAc/hexane gave 10(4a) (190
mg, 0.47 mmol, 95% yield) and 10(4b) (192 mg, 0.48 mmol, 96% yield)
both as white foams.
[0144] 10(4a): R.sub.f=0.25 (20% EtOAc/hexane);
[.alpha.].sub.D.sup.20 -37.0 (c 1.0, CHCl.sub.3); .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.70-7.60 (2H, m), 7.40-7.10 (8H, m), 6.63
(1H, d, J=2.2 Hz), 5.90-5.83 (1H, m), 5.51 (1H, dd, J=10.0 Hz, 1
Hz), 4.64 (1H, d, J=14.7 Hz), 4.38 (1H, d, J=14.7 Hz), 3.34 (1H,
dd, J=5.4 Hz, 3.3 Hz), 3.10-2.98 (1H, m), 2.85-2.65 (2H, m), 2.69
(1H, d, J=2.2 Hz), 2.46-2.38 (1H, m), 1.98-1.74 (2H, m); .sup.13C
NMR (100 MHz, CDCl.sub.3) .delta. 172.6, 169.0, 137.3, 136.5,
129.1, 128.6, 128.5 (2), 128.4 (2), 127.9 (2), 127.8 (2), 127.6,
127.2, 80.5, 75.5, 65.0, 50.1, 43.8, 42.5, 41.1, 34.2, 27.3, 23.6;
IR (neat) 2122, 1737, 1630, 1356 cm.sup.-1.
[0145] 10(4b): R.sub.f=0.20 (20% EtOAc/hexane);
[.alpha.].sub.D.sup.20 -15.0 (c 1.0, CHCl.sub.3); .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.70-7.60 (2H, m), 7.40-7.15 (8H, m), 6.62
(1H, d, J=2.2 Hz), 5.85-5.78 (1H, m), 5.50 (1H, dd, J=10.0 Hz, 1.6
Hz), 4.75 (1H, d, J=14.7 Hz), 4.36 (1H, d, I=14.7 Hz), 3.47 (1H,
dd, J=5.4 Hz, 3.2 Hz), 3.10-3.00 (2H, m), 2.85-2.60 (2H, m), 2.65
(1H, d, J=2.2 Hz), 2.48-2.28 (2H, m), 2.05-1.95 (1H, m), 1.82-1.77
(1H, m); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 172.6, 169.1,
137.2, 137.0, 129.0, 128.7, 128.5 (2), 128.4 (2), 127.9 (2), 127.8,
127.7 (2), 127.2, 80.5, 75.3, 65.3, 50.3, 43.8, 42.8, 41.4, 34.3,
27.3, 23.6; IR (neat) 2123, 1734, 1630, 1356 cm.sup.-1.
[0146] To a suspension of CuCN (134 mg, 1.5 mmol) in dry diethyl
ether (10 mL), MeLi (1.0 mL, 1.5 mmol) was added slowly at
0.degree. C. and stirred for 10 min when a clear homogeneous
solution formed. Then a solution of 10(4a) (190 mg, 0.47 mmol) in
dry diethyl ether was added to the reaction at 0.degree. C. The
reaction was warmed to the room temperature and stirred for 1 hour.
The reaction was quenched with saturated NH.sub.4Cl solution (5 mL)
and acidified with a solution of HCl (1 mL, 10%) at room
temperature. The reaction mixture was extracted with EtOAc (20
mL.times.3) and the combined organic layers were dried over
anhydrous MgSO.sub.4. Evaporation of solvent followed by
chromatography with 50% EtOAc/hexane gave 2(4a) (104 mg, 0.36 mmol,
74% yield) as white solid. [.alpha.].sub.D.sup.20 -21.4 (c 0.5,
CHCl.sub.3).
[0147] To a suspension of CuCN (134 mg, 1.5 mmol) in dry diethyl
ether (10 mL), MeLi (1.0 mL, 1.5 mmol) was added slowly at
0.degree. C. and stirred for 10 min when a clear homogeneous
solution formed. Then a solution of 10(4b) (192 mg, 0.48 mmol) in
dry diethyl ether was added to the reaction at 0.degree. C. The
reaction was warmed to the room temperature and stirred for 1 hour.
The reaction was quenched with saturated NH.sub.4Cl solution (5 mL)
and acidified with a solution of HCl (1 mL, 10%) at room
temperature. The reaction mixture was extracted with EtOAc (20
mL.times.3) and the combined organic layers were dried over
anhydrous MgSO.sub.4. Evaporation of solvent followed by
chromatography with 50% EtOAc/hexane gave 2(4b) (112 mg, 0.39 mmol,
83% yield) as white solid. [.alpha.].sub.D.sup.20 +22.0 (c 0.5,
CHCl.sub.3).
Synthesis of Enantio-Enriched
n-Alkyl-Octahydroisoquinolin-1-One-8-Carboxamides
[0148] The following enantio-enriched
n-alkyl-octahydroisoquinolin-1-one-8-carboxamides were prepared
from 2(4a) and 2(4b) following identical conditions reported for
syntheses of the racemic carboxamides.
##STR00060##
[0149] 11789: [.alpha.].sub.D.sup.20 +23.6 (c 1.0,
CHCl.sub.3)..sup.3
[0150] 11790: [.alpha.].sub.D.sup.20 -23.2 (c 1.0, CHCl.sub.3);
m.p. 180-182.degree. C.; (neat) 2867, 1695, 1614, 1482, 1315
cm.sup.-1; HRMS (ESI) m/z calcd for
C.sub.24H.sub.23ClF.sub.3N.sub.2O.sub.2 ([M+H].sup.+), 463.1400.
found 463.1404..sup.1,2,3
[0151] 11791: [.alpha.].sub.D.sup.20 +15.8 (c 1.0, CHCl.sub.3).
[0152] 11792: [.alpha.].sub.D.sup.20 -16.0 (c 1.0, CHCl.sub.3); IR
(neat) 2853, 1635, 1535, 1490, 1353 cm.sup.-1; .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.42 (1H, bs), 7.32-7.12 (5H, m),
5.88-5.80 (1H, m), 5.54 (1H, d, J=10.0 Hz), 4.64 (1H, d, J=14.8
Hz), 4.50 (1H, d, J=14.8 Hz), 3.83-3.70 (1H, m), 3.20 (1H, dd,
J=2.5 Hz, 2.5 Hz), 3.08 (1H, d, J=8.3 Hz), 3.07 (1H, d, J=8.5 Hz),
2.72-2.65 (2H, m), 2.40-2.25 (2H, m), 2.00-1.10 (12H, m); .sup.13C
NMR (100 MHz, CDCl.sub.3) .delta. 173.1, 170.8, 136.9, 128.8, 128.5
(2), 128.3, 127.8 (2), 127.3, 50.4, 48.1, 45.2, 44.4, 42.5, 35.7,
32.9 (2), 27.2, 25.8, 25.7, 24.9; HRMS (ESI) m/z calcd for
C.sub.23H.sub.31N.sub.2O.sub.2 ([M+H].sup.+), 367.2385. found
367.2387..sup.1,2
[0153] 11793: [.alpha.].sub.D.sup.20 +24.8 (c 1.0, CHCl.sub.3).
[0154] 11794: [.alpha.].sub.D.sup.20 -25.2 (c 0.6, CHCl.sub.3); IR
(neat) 2868, 1667, 1624, 1596, 1490 cm.sup.-1; HRMS (ESI) m/z calcd
for C.sub.23H.sub.25N.sub.2O.sub.2 ([M+H].sup.+), 361.1916. found
361.1911..sup.1,2
[0155] 11869*: [.alpha.].sub.D.sup.20 -20.5 (c 1.0, CHCl.sub.3);
m.p. 156-158.degree. C.; IR (neat) 2925, 1675, 1618, 1478, 1320
cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 11.83 (1H,
bs), 7.98 (1H, d, J=2.6 Hz), 7.74 (1H, dd, J=8.7 Hz, 2.5 Hz), 7.59
(1H, d, J=8.7 Hz), 7.34-7.15 (5H, m), 5.95-5.88 (1H, m), 5.58 (1H,
d, J=10.0 Hz, 1.2 Hz), 4.82 (1H, d, J=14.8 Hz), 4.45 (1H, d, J=14.8
Hz), 3.23-2.82 (5H, m), 2.52-2.45 (2H, m), 2.03-1.80 (2H, m);
.sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 172.9, 171.5, 138.5,
136.3, 135.1, 129.4, 128.7 (2), 128.1, 127.6 (2), 123.9, 119.2 (q),
50.8, 48.6, 44.3, 42.2, 36.2, 27.2, 26.6..sup.1,2,4
##STR00061##
[0156] 11795: m.p. 200-202.degree. C.; IR (neat) 2931, 1694, 1612,
1540, 1482 cm.sup.-1; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
10.02 (1H, bs), 8.12 (1H, J=1.7 Hz), 7.68 (1H, d, J=8.6 Hz),
7.41-7.15 (6H, m), 6.10-6.00 (1H, m), 5.68 (1H, d, J=10.1 Hz), 4.66
(2H, s), 3.81-3.67 (1H, m), 3.40-1.80 (8H, m); .sup.13C NMR (100
MHz, CDCl.sub.3) .delta. 172.9, 170.6, 137.8, 136.5, 131.6, 128.9,
128.7 (2), 127.6, 127.5, 127.2, 127.1 (2), 123.5, 118.6 (q), 51.1,
44.7, 42.8, 41.5, 29.8, 27.0, 23.6.
[0157] For enantiomerically pure compounds: [0158] 1. 11789 and
11790 are the enantiomeric pairs of racemic 8620. 11791 and 11792
are the enantiomeric pairs of racemic 8920, 11793 and 11794 are the
enantiomeric pairs of racemic 8937. Compound 11869* is a single
enantiomer of racemic 11869 and has the absolute configuration
shown. [0159] 2. Enantiomeric excess (ee) for 11789 and 11790 were
found to be 95% and 94% respectively using analytical HPLC
(Chiralcel OD-H, 1 mL/min, 10% i-PrOH/hexane). [0160] 3. The
absolute configuration of 11869* was confirmed by X-Ray
crystallography using anomalous dispersion. This also confirms the
absolute configuration of all the carboxamides listed above. [0161]
4. The individual enantiomers can be separated from each other or
combined into a racemic mixture. It is recognized that some pure
enantiomers may be more biologically active compared to other
enantiomers or the racemic mixture. The disclosure of one
enantiomer is considered to disclose all of its other
enantiomers.
[0162] The compounds prepared herein were tested for bioactivity
with opioid receptors. Agonist assays: Data represent the percent
efficacy (relative to cognate agonist) of the test compound at 10
micromolar. The response to a saturating concentration of a
reference antagonist or to vehicle (usually none or negligible) is
set to 0%. Antagonist assays: Data represent the percent inhibition
of the response to an EC90 concentration (empirically determined
immediately prior to assay) of cognate/reference agonist by the
test compound at 10 micromolar. The response inhibition elicited by
a saturating concentration of reference antagonist is set to 100%;
the response inhibition (usually none or negligible) elicited by
vehicle is set to 0%. The following tables provide data related to
the bioactivity for opioid receptors of the compounds described
herein.
TABLE-US-00005 TABLE 4 Opioid secondary binding activity for
Formula 10 analogs DOR, Ki KOR, Ki MOR, Ki C# n= R1 R2 R3 (nM) (nM)
(nM) 8909 1 None --CH2CH2OCH2CH2-- cyclopropyl 8910 1 none
--CH2CH2OCH2CH2-- cyclohexyl 2950 8911 1 none --CH2CH2OCH2CH2--
n-butyl 8912 1 none --CH2CH2N(Ph)CH2CH2-- benzyl 3854 4803 2396
8913 1 none --CH2CH2N(Ph)CH2CH2-- cyclopropyl 3194 8914 1 none
--CH2CH2N(Ph)CH2CH2-- n-butyl 8915 1 none --CH2CH2N(Ph)CH2CH2--
3,4-dimethoxybenzyl 4693 8897 5536 8916 1 H _CH2CH2Ph benzyl 561
4599 8917 1 H 2-phenylethyl cyclopropyl 8918 1 H 2-phenylethyl
n-butyl 3271 8919 1 H n-butyl n-butyl 2813 8920 1 H cyclohexyl
benzyl 298 8921 1 H cyclohexyl cyclopropyl 8922 1 H cyclohexyl
cyclohexyl 167 8923 1 H cyclohexyl n-butyl 1029 8924 1 H
4-methoxybenzyl benzyl 3729 8925 1 H 4-methoxybenzyl cyclopropyl
8926 1 H 4-methoxybenzyl cyclohexyl 2691 8927 1 H 4-methoxybenzyl
n-butyl 8928 1 H 4-methoxybenzyl 3,4-dichlorobenzyl 964 7395 8929 1
H 4-methoxybenzyl 3,4-dimethoxybenzyl 8930 1 H pyridln-3-yl-methyl
cyclopropyl 8931 1 H pyridin-3-yl-methyl cyclohexyl 552 8932 1 H
pyridin-3-yl-methyl 3,4-dichlorobenzyl 693 8933 1 H thiazole benzyl
200 8934 1 H thiazole cyclopropyl 8935 1 H thiazole cyclohexyl 160
8936 1 H thiazole n-butyl 355 8937 1 H phenyl benzyl 850 146 2574
8938 1 H phenyl cyclopropyl 1404 8939 1 H phenyl cyclohexyl 8938 71
8940 1 H phenyl n-butyl 6109 677 8941 1 H phenyl 3,4-dichlorobenzyl
1112 259 2697 8942 1 H phenyl 3,4-dimethoxybenzyl 4607 3058 3523
8943 1 H 4-methoxyphenyl benzyl 883 8944 1 H 4-methoxyphenyl
cyclopropyl 8945 1 H 4-methoxyphenyl cyclohexyl 5669 1790 8946 1 H
4-methoxyphenyl n-butyl 5036 8947 1 H 4-methoxyphenyl
3,4-dichlorobenzyl 2558 3493 8948 1 H 2,4-dichlorophenyl
cyclopropyl 8949 1 H 2,4-dichlorophenyl cyclohexyl 506 8950 1 H
2,4-dichlorophenyl n-butyl 1139 8951 1 H 2,4-dichlorophenyl
3,4-dichlorobenzyl 2443 8952 1 H 2,4-dichlorophenyl
3,4-dimethoxybenzyl 5660 8620 1 H 4-chloro-3- benzyl 5.3 (Avg)
3,552 trifluoromethylphenyl 8954 1 H 4-chloro-3- cyclopropyl 493
trifluoromethylphenyl 8955 1 H 4-chloro-3- cyclohexyl 111
trifluoromethylphenyl 8956 1 H 4-chloro-3- n-butyl 190 2220
trifluoromethylphenyl 8957 1 H 4-chloro-3- 3,4-dichlorobenzyl
trifluoromethylphenyl 8958 1 H 4-chloro-3- 3,4-dimethoxybenzyl 2540
trifluoromethylphenyl 5084 1 H 2-phenylethyl 3,4-dichlorobenzyl
4802 5189 5085 1 H 3,4-difluorophenyl 3,4-dichlorobenzyl 2961 4655
5086 1 H piperidine 3,4-dichlorobenzyl 8599 1 H thiazole
3,4-dichlorobenzyl 8600 1 OH cyclopropyl 10821 1 H 3-chlorophenyl
phenyl 420 10822 1 H 4-trifluoromethylphenyl phenyl 786 10823 1 H
3,4-difluorophenyl phenyl 375 10824 1 H 2,6-difluorophenyl phenyl
5001 10825 1 H 4-chloro-3- phenyl 100 trifluoromethylphenyl 10826 1
H 2,4-dichlorophenyl phenyl 287 10827 1 H 4-methoxyphenyl
2-phenylethyl 10828 1 H 3-chlorophenyl 2-phenylethyl 10829 1 H
4-trifluoromethylphenyl 2-phenylethyl 10830 1 H
3-trifluoromethylphenyl 2-phenylethyl 10831 1 H 3,4-difluorophenyl
2-phenylethyl 10832 1 H 2,6-difluorophenyl 2-phenylethyl 10833 1 H
4-chloro-3- 2-phenylethyl trifluoromethylphenyl 10834 1 H
2,4-dichlorophenyl 2-phenylethyl 6485 10835 1 H 4-methoxyphenyl
4-chlorobenzyl 5074 10836 1 H 3-chlorophenyl 4-chlorobenzyl 225
10837 1 H 4-trifluoromethylphenyl 4-chlorobenzyl 2533 10838 1 H
3-trifluoromethylphenyl 4-chlorobenzyl 1512 10839 1 H
3,4-difluorophenyl 4-chlorobenzyl 652 10840 1 H 2,6-difluorophenyl
4-chlorobenzyl 10841 1 H 4-chloro-3- 4-chlorobenzyl 465
trifluoromethylphenyl 10842 1 H 2,4-dichlorophenyl 4-chlorobenzyl
10843 1 H 4-methoxyphenyl isopropyl 10844 1 H 3-chlorophenyl
isopropyl 2131 10845 1 H 4-trifluoromethylphenyl isopropyl 10846 1
H 3-trifluoromethylphenyl isopropyl 621 10847 1 H
3,4-difluorophenyl isopropyl 4197 10848 1 H 2,6-difluorophenyl
isopropyl 10849 1 H 4-chloro-3- isopropyl 261 trifluoromethylphenyl
10850 1 H 2,4-dichlorophenyl isopropyl 2409 10851 1 H
4-trifluoromethylphenyl 4-trifluoromethylbenzyl 10852 1 H
3-trifluoromethylphenyl 4-trifluoromethylbenzyl 10853 1 H
3,4-difluorophenyl 4-trifluoromethylbenzyl 10854 1 H
2,6-difluorophenyl 4-trifluoromethylbenzyl 11476 1 H (desoxo)
phenyl (desoxo) 3,4- 1697 1,108 dichlorobenzyl 11477 1 H (desoxo)
4-chloro-3- (desoxo) benzyl 546 338 trifluoromethylphenyl 11478 1 H
4-chloro-3- benzyl 51 1,748 (olefin trifluoromethylphenyl
hydrogenated) 11479 1 H 4-chloro-3- cyclohexyl 792 (olefin
trifluoromethylphenyl hydrogenated) 11789 1 H 4-chloro-3- benzyl
132 3477 trifluoromethylphenyl 11790 1 H 4-chloro-3- benzyl 3.2 208
trifluoromethylphenyl 11791 1 H cyclohexyl benzyl 1303 11792 1 H
cyclohexyl benzyl 213 11793 1 H phenyl benzyl 1605 11794 1 H phenyl
benzyl 137 1657 11795 1 H 4-chloro-3- benzyl 158 5424 (epi
trifluoromethylphenyl scaffold) 11808 1 ".dbd.R2" (side chain 1)
4-chlorobenzyl 2293 11809 1 H (side chain 2) 4-chlorobenzyl 11810 1
H (side chain 3) 4-chlorobenzyl 11811 1 H (side chain 4)
4-chlorobenzyl 11812 1 H (side chain 5) 4-chlorobenzyl 11813 1 H
(side chain 6) 4-chlorobenzyl 1157 1909 11814 1 ".dbd.R2" (side
chain 1) benzyl 11815 1 H (side chain 2) benzyl 11816 1 H (side
chain 3) benzyl 2634 11817 1 H (side chain 4) benzyl 11818 1 H
(side chain 5) benzyl 6168 11819 1 H (side chain 6) benzyl 1290
11821 1 H (side chain 2) cyclohexyl 4708 11822 1 H (side chain 3)
cyclohexyl 4499 11823 1 H (side chain 4) cyclohexyl 11824 1 H (side
chain 5) cyclohexyl 11825 1 H (side chain 6) cyclohexyl 1106 11826
1 ".dbd.R2" (side chain 1) phenethyl 11827 1 H (side chain 2)
phenethyl 11828 1 H (side chain 3) phenethyl 11829 1 H (side chain
4) phenethyl 11830 1 H (side chain 5) phenethyl 11831 1 H (side
chain 6) phenethyl 1525 11836 1 H (.+-.)-.alpha.-methylbenzyl
cyclohexyl 599 11837 1 H 2-chloro-5- cyclohexyl 8021 94
trifluoromethylphenyl 11848 1 H (.+-.)-.alpha.-methylbenzyl
2-phenylethyl 3488 11849 1 H 2-chloro-5- 2-phenylethyl 1816 2406
trifluoromethylphenyl 11854 1 H (.+-.)-.alpha.-methylbenzyl benzyl
712 11856 1 H 4-chloro-3- cyclohexyl 531 trifluoromethylbenzyl
11857 1 H 4-bromo-3- cyclohexyl 814 trifluoromethylphenyl 11858 1 H
4-chloro-2- cyclohexyl 866 trifluoromethylphenyl 11859 1 H
2-chloro-4- cyclohexyl trifluoromethylphenyl 11860 1 H
4-chloro-3-methylphenyl cyclohexyl 716 11862 1 H 4-chloro-3- phenyl
98 trifluoromethylbenzyl 11863 1 H 4-bromo-3- phenyl 12
trifluoromethylphenyl 11864 1 H 4-chloro-2- phenyl 68 (Avg)
trifluoromethylphenyl 11865 1 H 2-chloro-4- phenyl 180
trifluoromethylphenyl 11866 1 H 4-chloro-3-methylphenyl phenyl 33.5
(Avg) 11868 1 H 4-chloro-3- benzyl 455 2165 trifluoromethylbenzyl
11869 1 H 4-bromo-3- benzyl 16.5 (Avg) 648 trifluoromethylphenyl
11872 1 H 4-chloro-3-methylphenyl benzyl 86 (Avg) 2179 11874 1 H
4-chloro-3- 2-phenylethyl 319 1244 trifluoromethylbenzyl 11875 1 H
4-bromo-3- 2-phenylethyl 2283 1177 trifluoromethylphenyl 11876 1 H
4-chloro-3- 2-phenylethyl trifluoromethylphenyl 11877 1 H
2-chloro-4- 2-phenylethyl 3303 trifluoromethylphenyl 11878 1 H
4-chloro-3-methylphenyl 2-phenylethyl 4116 1674 11886 1 H
cyclohexyl cyclohexylmethyl 177 11887 1 H phenyl cyclohexylmethyl
187 11888 1 H 3-chlorophenyl cyclohexylmethyl 68.0 (AVE) 11889 1 H
3,4-difluorophenyl cyclohexylmethyl 131 11890 1 H 4-chloro-3-
cyclohexylmethyl 840 trifluoromethylphenyl 11891 1 H thiazole
cyclohexylm ethyl 183 11892 1 H cyclohexyl H 11893 1 H phenyl H
11894 1 H 3-chlorophenyl H 11895 1 H 3,4-difluorophenyl H 11896 1 H
4-chloro-3- H 729 trifluoromethylphenyl 11897 1 H thiazole H 11899
1 H (side chain 7) phenyl 11901 1 H (side chain 8) benzyl 11902 1 H
(side chain 7) benzyl 3375 11903 1 ".dbd.R2" (side chain 9) benzyl
11904 1 H (side chain 10) 3,4-dimethoxybenzyl 8766 11905 1
".dbd.R2" (side chain 1) 3,4-dimethoxybenzyl 11908 1 H (side chain
7) cyclohexyl 11909 1 H 4-fluorophenyl benzyl 276 11916 1 H methyl
cyclohexyl 11918 1 H benzyl cyclohexyl 358 11919 1 H
3-chloro-4-methylphenyl cyclohexyl 102 11920 1 H 4-nitro-3-
cyclohexyl 5108 trifluoromethylphenyl 11921 1 H 3-pyridine-yl
cyclohexyl 836 11922 1 H methyl phenyl 11923 1 cyclohexyl
cyclohexyl phenyl 11925 1 H 3-chloro-4-methylphenyl phenyl 15 11926
1 H 4-nitro-3- phenyl 35 trifluoromethylphenyl 11927 1 H
3-pyridine-yl phenyl 2096 11928 1 H methyl benzyl 11929 1
cyclohexyl cyclohexyl benzyl 11930 1 H benzyl benzyl 739 11931 1 H
3-chloro-4-methylphenyl benzyl 61 2574 11932 1 H 4-nitro-3- benzyl
41 200 trifluoromethylphenyl 11933 1 H 3-pyridine-yl benzyl 3526
11940 1 H adamantyl cyclopropylmethyl 701 11941 1 H 4-nitrophenyl
cyclopropylmethyl 11942 1 H 4-iodophenyl cyclopropylmethyl 1276
11943 1 H 3-bromobenzyl cyclopropylmethyl 11944 1 H 4-methylbenzyl
cyclopropylmethyl 11945 1 H 2,4,6-trimethylphenyl cyclopropylmethyl
11946 1 H adamantyl phenyl >10,000 116 11947 1 H 4-nitrophenyl
phenyl 141 11948 1 H 4-iodophenyl phenyl 83 11949 1 H 3-bromobenzyl
phenyl 302 11950 1 H 4-methylbenzyl phenyl 297 11951 1 H
2,4,6-trimethylphenyl phenyl 4300 11952 1 H adamantyl benzyl 11953
1 H 4-nitrophenyl benzyl 11954 1 H 4-iodophenyl benzyl 194 2825
11955 1 H 3-bromobenzyl benzyl 438 3275 11956 1 H 4-methylbenzyl
benzyl 2362 11957 1 H 2,4,6-trimethylphenyl benzyl 3216 12084 1
".dbd.R2" _CH2CH2CH2CH2.sub.-- cyclohexyl 6292 12085 1 H
cyclopropylmethyl cyclohexyl 475 12086 1 methyl methyl cyclohexyl
>10,000
12087 1 H 4-chlorophenyl cyclohexyl 582 12088 1 H
3,4,5-trifluorophenyl cyclohexyl 2793 12089 1 ".dbd.R2"
_CH2CH2CH2CH2.sub.-- cyclopropylmethyl 12090 1 H cyclopropylmethyl
cyclopropylmethyl 5650 12091 1 methyl methyl cyclopropylmethyl
12092 1 H 4-chlorophenyl cyclopropylmethyl 3234 12093 1 H
3,4,5-trifluorophenyl cyclopropylmethyl 4370 12094 1 ".dbd.R2"
_CH2CH2CH2CH2.sub.-- phenyl >10,000 12095 1 H cyclopropylmethyl
phenyl 1225 12096 1 methyl methyl phenyl >10,000 12097 1 H
4-chlorophenyl phenyl 168 12098 1 H 3,4,5-trifluorophenyl phenyl
289 12099 1 ".dbd.R2" _CH2CH2CH2CH2.sub.-- benzyl 6647 12100 1 H
cyclopropylmethyl benzyl 539 12101 1 methyl methyl benzyl
>10,000 12102 1 H 4-chlorophenyl benzyl 165 5029 12103 1 H
3,4,5-trifluorophenyl benzyl 180 12110 1 H allyl cyclohexylmethyl
1428 12111 1 H anthranilamide cyclohexylmethyl 12112 1 H 4-(ethyl
carboxylate)phenyl cyclohexylmethyl 12113 1 methyl phenyl
cyclohexylmethyl 2830 12114 1 H 3,4-dimethoxybenzyl
cyclohexylmethyl 12115 1 ".dbd.R2" ethyl pipecolinate
cyclohexylmethyl 9155 12116 1 H allyl phenyl 12117 1 H
anthranilamide phenyl 648 12118 1 H 4-(ethyl carboxylate)phenyl
phenyl 12119 1 methyl phenyl phenyl 4014 12120 1 H
3,4-dimethoxybenzyl phenyl 12121 1 H ethyl pipecolinate phenyl
12122 1 H allyl benzyl 3431 12123 1 H anthranilamide benzyl 5932
12124 1 H 4-(ethyl carboxylate)phenyl benzyl 7821 12125 1 H phenyl
benzyl >10,000 12126 1 H 3,4-dimethoxybenzyl benzyl >10,000
12127 1 ".dbd.R2" ethyl pipecolinate benzyl >10,000 12134 1
isopropyl cyclohexyl cyclohexylmethyl 1634 12135 1 ".dbd.R2"
_CH2CH2(NMe)CH2CH2.sub.-- cyclohexylmethyl 12136 1 H
3,4-dichlorobenzyl cyclohexylmethyl 1301 12137 1 H 4-hexylphenyl
cyclohexylmethyl 12138 1 H pyrazine-2-yl cyclohexylmethyl 1651
12140 1 isopropyl cyclohexyl phenyl 416 12141 1 ".dbd.R2"
_CH2CH2(NMe)CH2CH2.sub.-- phenyl 12142 1 H 3,4-dichlorobenzyl
phenyl 103 12143 1 H 4-hexylphenyl phenyl 12144 1 H pyrazine-2-yl
phenyl 12147 1 ".dbd.R2" --CH2CH2(NMe)CH2CH2-- benzyl 12148 1 H
3,4-dichlorobenzyl benzyl 644 2779 12149 1 H 4-hexylphenyl benzyl
12150 1 H pyrazine-2-yl benzyl 12152 1 H cyclohexyl
cyclopropylmethyl 2518 12153 1 H phenyl cyclopropylmethyl 6624
12154 1 H 3-chlorophenyl cyclopropylmethyl 1177 12155 1 H
3,4-difluorophenyl cyclopropylmethyl 2252 12156 1 H 4-chloro-3-
cyclopropylmethyl 788 trifluoromethylphenyl 12157 1 H thiazole
cyclopropylmethyl 3501 12160 1 methyl phenyl cyclohexyl 2978 12161
1 H 4-chloro-3- (4'-methoxybiphenyl- 2126 trifluoromethylphenyl
3-yl)methyl 12163 1 H cyclohexyl (.+-.)-.alpha.-methylbenzyl 1162
5909 12164 1 H phenyl (.+-.)-.alpha.-methylbenzyl 1384 4775 12165 1
H 3-chlorophenyl (.+-.)-.alpha.-methylbenzyl 1207 12166 1 H
3,4-difluorophenyl (.+-.)-.alpha.-methylbenzyl 3203 12168 1 H
thiazole (.+-.)-.alpha.-methylbenzyl 1450 12170 1 H 4-chloro-3-
3-bromobenzyl 1718 1462 trifluoromethylphenyl 12171 1 H 4-bromo-3-
3-bromobenzyl 503 694 trifluoromethylphenyl 12172 1 H 4-bromo-3-
cyclopropylmethyl 1282 trifluoromethylphenyl 12173 1 H 4-bromo-3-
cyclohexylmethyl 508 trifluoromethylphenyl 13474 1 H 4-fluoro-3-
benzyl 192 trifluoromethylphenyl 13475 1 H 4-methyl-3- benzyl 39
1163 trifluoromethylphenyl 13476 1 H 2,3-difluoro-4-methylphenyl
benzyl 559 13478 1 H 4-bromo-3- 2-fluorobenzyl 22 299
trifluoromethylphenyl 13479 1 H 4-methyl-3- 2-fluorobenzyl 9.5 763
trifluoromethylphenyl 13480 1 H 4-bromo-3- 2-methoxybenzyl
trifluoromethylphenyl 13481 1 H 4-methyl-3- 2-methoxybenzyl 1342
1898 trifluoromethylphenyl 13482 1 H 4-chloro-3- 2-fluoro-6-
trifluoromethylphenyl methoxybenzyl 13483 1 H 4-bromo-3-
2-fluoro-6- trifluoromethylphenyl methoxybenzyl 13484 1 H
4-methyl-3- 2-fluoro-6- trifluoromethylphenyl methoxybenzyl 13485 1
H 2,3-difluoro-4-methylphenyl 2-fluoro-6- 5122 methoxybenzyl 14309
1 H pentafluorophenyl benzyl 2969 14417 2 H phenyl benzyl 14418 2 H
cyclohexyl benzyl 14419 2 H 3-chlorophenyl benzyl 14420 2 H
4-bromo-3- benzyl trifluoromethylphenyl 14421 2 H 4-chloro-3-
benzyl trifluoromethylphenyl 14422 2 H thiazole benzyl 14423 2 H
phenyl cyclohexyl 8836 14424 2 H cyclohexyl cyclohexyl 1690 14425 2
H 3-chlorophenyl cyclohexyl 6044 14426 2 H 4-bromo-3- cyclohexyl
trifluoromethylphenyl 14427 2 H 4-chloro-3- cyclohexyl
trifluoromethylphenyl 14428 2 H thiazole cyclohexyl 2990 14429 0 H
phenyl benzyl 7019 14430 0 H cyclohexyl benzyl 14431 0 H
3-chlorophenyl benzyl 14432 0 H 4-bromo-3- benzyl 2767
trifluoromethylphenyl 14433 0 H 4-chloro-3- benzyl 1938
trifluoromethylphenyl 14434 0 H thiazole benzyl 14435 0 H phenyl
cyclohexyl 1608 14436 0 H cyclohexyl cyclohexyl 2150 14437 0 H
3-chlorophenyl cyclohoxyl 962 14438 0 H 4-bromo-3- cyclohexyl 1918
trifluoromethylphenyl 14439 0 H 4-chloro-3- cyclohexyl 5348
trifluoromethylphenyl 14440 0 H thiazole cyclohexyl 1038 14827 1
methyl methoxy 2-phenylethyl 14828 1 H biphenyl-2-yl phenyl 14829 1
H biphenyl-2-yl benzyl 14830 1 H phenyl (4'-methylbiphenyl-
3-yl)methyl 14832 1 H 4-chloro-3- (4'-methylbiphenyl-
trifluoromethylphenyl 3-yl)methyl 14833 1 H 4-chloro-3-
(4'-trifluoromethoxy- trifluoromethylphenyl biphenyl-3-yl)methyl
14834 1 H phenyl (side chain 11) 14835 1 H phenyl (side chain 12)
14836 1 H phenyl (side chain 13) 14837 1 H (4'-methylbiphenyl-3-
phenyl yl)methyl 14838 1 H (4'-methylbiphenyl-3- benzyl yl)methyl
14839 1 H (4'-carbamoylbiphenyl-3- benzyl yl)methyl 14840 1 H (side
chain 16) benzyl 14841 1 H (side chain 11) benzyl 14842 1 H (side
chain 15) benzyl 14843 1 H (side chain 14) benzyl 14844 1 H
3-(thiophen-2-yl)benzyl benzyl 14845 1 H (side chain 13) benzyl (oh
= olefin hydrogenated) (epi = epi scaffold or unknown epimer) (when
R1 is nothing, then R2 is a ring) (Value for C# 8620 is an
average)
TABLE-US-00006 TABLE 5 Example compounds with KOR/MOR binding data.
##STR00062## ##STR00063## ##STR00064## ##STR00065## ##STR00066##
##STR00067## ##STR00068## ##STR00069## ##STR00070## ##STR00071##
##STR00072## ##STR00073## ##STR00074## ##STR00075## ##STR00076##
##STR00077## ##STR00078## ##STR00079## ##STR00080## ##STR00081##
##STR00082## ##STR00083## ##STR00084## ##STR00085##
##STR00086##
[0163] Table 6 provides data that represent the average percent
efficacy (N=4; relative to cognate agonist) of the test compound at
10 micromolar concentration. The response to a saturating
concentration of cognate/reference agonist is set to 100%; the
response to a saturating concentration of a reference antagonist or
to vehicle (usually none or negligible) is set to 0%. All
functional assays are performed on stably transfected CHO-cell
lines expressing hKOR, hMOR or hDOR.
[0164] Experimental Procedure and Data Analysis: Assay buffer: 50
mM HEPES, 5 mM MgCl2, 150 mM NaCl, 0.2 mM EDTA, 100 mg/1 ascorbic
acid, pH 7.4 Reference agonist (U69593 for KOR; DAMGO for MOR and
DADL for DOR) and compounds to be tested are dissolved in assay
buffer or DMSO according to solubility. Serial dilutions of the
test and reference compounds are made in binding buffer at 2.times.
assay concentration (final assay concentrations ranging from 0.1 nM
to 10 .mu.M). Crude membrane fractions (1.8 to 3.5 cm2/well)
(prepared from 10-cm plates by harvesting PBS-rinsed monolayers,
resuspending and lysing in chilled, hypotonic 50 mM Tris-HCl, pH
7.4, centrifuging at 20,000.times.g, decanting the supernatant and
storing at -80 degrees centigrade; typically, one 10-cm plate
provides sufficient material for 24 wells) are resuspended in 1.2
ml of assay buffer containing 20 .mu.M GDP, wheat germ agglutinin
(WGA)-coated scintillation proximity beads (2.4 mg), and
[35S]GTP.gamma.S (300 pM final). The suspension is then added (50
.mu.l/well) to 50 .mu.l of the 2.times. test or reference compounds
(each concentration assayed in triplicate) in flexible transparent
PET 96-well plates. The reaction plate is sealed and incubated for
90 min at room temperature, then centrifuged for 5 min at
216.times.g, and finally loaded into a Wallac MicroBeta TriLux
counter. Non-specific [35S]GTP.gamma.S binding is assessed at the
maximum concentration of reference agonist in the presence of 10
.mu.M antagonist. The background signal is measured at 10 .mu.M
reference agonist in the presence of 10 .mu.M unlabeled
GTP.gamma.S. Raw data (dpm) representing total [35S]GTP.gamma.S
binding (i.e., specific+non-specific binding) are calculated as a
percentage of the maximum response by the reference agonist (in
this case U69593).
TABLE-US-00007 TABLE 6 Compounds having KOR agonist activity. C#
KOR agonist 8909 7.8 8910 42.7 8911 8.3 8912 8 8913 27.5 8914 9.9
8915 6.7 8916 110.6 8917 6.6 8918 12.6 8919 40.3 8920 94.2 8921 4.1
8922 8 8923 67.6 8924 18.8 8925 5.4 8926 29 8927 8.7 8928 17.7 8929
3.6 8930 4 8931 67.7 8932 74.6 8933 121.5 8934 11 8935 129.2 8936
93.8 8937 136.6 8938 21.8 8939 133.7 8940 112.1 8941 79 8942 5.4
8943 72.9 8944 1 8945 54.3 8946 23.5 8947 17.3 8948 29.6 8949 67.9
8950 72.9 8951 23.2 8952 -2.6 8953 8954 66.4 8955 93.2 8956 118.7
8957 33.1
[0165] Table 7 shows compounds having KOR agonist activity.
[0166] Experimental Procedure and Data Analysis: Assay buffer: 50
mM HEPES, 5 mM MgCl2, 150 mM NaCl, 0.2 mM EDTA, 100 mg/l ascorbic
acid, pH 7.4 Reference agonist (U69593 for KOR; DAMGO for MOR and
DADL for DOR) and compounds to be tested are dissolved in assay
buffer or DMSO according to solubility. Serial dilutions of the
test and reference compounds are made in binding buffer at 2.times.
assay concentration (final assay concentrations ranging from 0.1 nM
to 10 .mu.M). Crude membrane fractions (1.8 to 3.5 cm2/well)
(prepared from 10-cm plates by harvesting PBS-rinsed monolayers,
resuspending and lysing in chilled, hypotonic 50 mM Tris-HCl, pH
7.4, centrifuging at 20,000.times.g, decanting the supernatant and
storing at -80 degrees centigrade; typically, one 10-cm plate
provides sufficient material for 24 wells) are resuspended in 1.2
ml of assay buffer containing 20 .mu.M GDP, wheat germ agglutinin
(WGA)-coated scintillation proximity beads (2.4 mg), and
[35S]GTP.gamma.S (300 pM final). The suspension is then added (50
.mu.l/well) to 50 .mu.l of the 2.times. test or reference compounds
(each concentration assayed in triplicate) in flexible transparent
PET 96-well plates. The reaction plate is sealed and incubated for
90 min at room temperature, then centrifuged for 5 min at
216.times.g, and finally loaded into a Wallac MicroBeta TriLux
counter. Non-specific [35S]GTP.gamma.S binding is assessed at the
maximum concentration of reference agonist in the presence of 10
.mu.M antagonist. The background signal is measured at 10 .mu.M
reference agonist in the presence of 10 .mu.M unlabeled
GTP.gamma.S.
[0167] Raw data (dpm) representing total [35S]GTP.gamma.S binding
(i.e., specific+non-specific binding) are plotted as a function of
the logarithm of the molar concentration of the drug (i.e., test or
reference compound). Non-linear regression of the normalized (i.e.,
fold increase in [35S]GTP.gamma.S binding over that observed in the
absence of test or reference compound) data is performed in Prism
4.0 (GraphPad Software) using the built-in three parameter logistic
model (i.e., sigmoidal concentration-response) describing
agonist-stimulated activation of one receptor population:
y=bottom+[(top-bottom)/(1+10.times.-log EC50)]
where bottom equals the best-fit basal [35S]GTP.gamma.S binding and
top equals the best-fit maximum [35S]GTP.gamma.S binding. The log
EC50 (i.e., the log of the drug concentration that increases
[35S]GTP.gamma.S binding by 50% of the top) is thus estimated from
the data, and the EC50 (agonist potency) is obtained. To obtain an
estimate of the relative efficacy of the test compound (Rel. Emax),
its data-fit top is compared to and expressed as a ratio of that
for the reference agonist (Rel. Emax of 1.00). Reference antagonist
for KOR was nor-BNI; for DOR was naltrindole and for MOR was
naloxone.
TABLE-US-00008 TABLE 7 Compounds having KOR agonist activity, with
values shown as ic50's (uM). KOR agonist C# EC.sub.50 values Emax
values relative to U69593 8620 4.7 67% 10825 26 82% 11790 12 78%
11834 839 71% 11837 419 106% 11862 238 81% 11863 10 95% 11864 503
119% 11866 417 123% 11869 158 68% 11872 1,431.00 123% 11888 868
70%
TABLE-US-00009 TABLE 8 Example ester compounds with KOR/MOR binding
data, where Ki is for the KOR. ##STR00087## ##STR00088##
##STR00089## ##STR00090## ##STR00091## ##STR00092## ##STR00093##
##STR00094## ##STR00095## ##STR00096## ##STR00097## ##STR00098##
##STR00099## ##STR00100## ##STR00101##
TABLE-US-00010 TABLE 9 Example lactone compounds with KOR binding
data ##STR00102## ##STR00103## ##STR00104## ##STR00105##
##STR00106## ##STR00107## ##STR00108## ##STR00109## ##STR00110##
##STR00111## ##STR00112## ##STR00113## ##STR00114## ##STR00115##
##STR00116## ##STR00117## ##STR00118## ##STR00119## ##STR00120##
##STR00121## ##STR00122## ##STR00123## ##STR00124## ##STR00125##
##STR00126## ##STR00127##
[0168] The present invention may be embodied in other specific
forms without departing from its spirit or essential
characteristics. The described embodiments are to be considered in
all respects only as illustrative and not restrictive. The scope of
the invention is, therefore, indicated by the appended claims
rather than by the foregoing description. All changes which come
within the meaning and range of equivalency of the claims are to be
embraced within their scope. All references recited herein are
incorporated herein by specific reference in their entirety.
* * * * *