U.S. patent application number 17/204549 was filed with the patent office on 2021-07-01 for orally active melanocortin receptor-4 compounds.
The applicant listed for this patent is Palatin Technologies, Inc.. Invention is credited to James Bullington, John Harold Dodd, Axel Metzger.
Application Number | 20210198201 17/204549 |
Document ID | / |
Family ID | 1000005491832 |
Filed Date | 2021-07-01 |
United States Patent
Application |
20210198201 |
Kind Code |
A1 |
Bullington; James ; et
al. |
July 1, 2021 |
Orally Active Melanocortin Receptor-4 Compounds
Abstract
A compound of the formula ##STR00001## where R.sub.1, R.sub.2,
R.sub.3, R.sub.4, R.sub.5, R.sub.6a, and R.sub.6b are as defined in
the specification and claims, or an enantiomer, stereoisomer or
diastereoisomer thereof, or a pharmaceutically acceptable salt
thereof, and the use thereof in the treatment of diseases,
disorders, syndromes and conditions responsive to modulation of a
melanocortin receptor.
Inventors: |
Bullington; James; (Hamilton
Square, NJ) ; Metzger; Axel; (Jackson, NJ) ;
Dodd; John Harold; (Spring Mills, PA) |
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Applicant: |
Name |
City |
State |
Country |
Type |
Palatin Technologies, Inc. |
Cranbury |
NJ |
US |
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|
Family ID: |
1000005491832 |
Appl. No.: |
17/204549 |
Filed: |
March 17, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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PCT/US2019/051423 |
Sep 17, 2019 |
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17204549 |
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62732680 |
Sep 18, 2018 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 45/06 20130101;
C07D 211/58 20130101 |
International
Class: |
C07D 211/58 20060101
C07D211/58; A61K 45/06 20060101 A61K045/06 |
Claims
1. A compound of formula I: ##STR00058## or an enantiomer,
stereoisomer or diastereoisomer thereof, or a pharmaceutically
acceptable salt thereof, wherein: R.sub.1 is --R.sub.7 or
--NR.sub.8R.sub.9; R.sub.2 and R.sub.3 are each independently a
C.sub.1 to C.sub.17 linear or branched alkyl, cycloalkyl,
alkylcycloalkyl, aryl, and alkylaryl, optionally substituted with
one or more substituents, and when one or more substituents are
present, such substituents are the same or different and
independently comprising oxo, carboxy, amino, monosubstituted
amino, disubstituted amino, or nitro, and when R.sub.2 and R.sub.3
are different, including all sterioisomers thereof, provided that
taken together R.sub.2 and R.sub.3 can form cycloalkyl, optionally
substituted, or alternatively that taken together one of R.sub.2
and R.sub.3 and one of R.sub.8 and R.sub.9 comprising a part of
R.sub.4 can form heterocycloalkyl, optionally substituted; R.sub.4
is --OH, --OR.sub.7, or --NR.sub.8R.sub.9; R.sub.5 is H, a C.sub.1
to C.sub.17 linear or branched alkyl, cycloalkyl, or
alkylcycloalkyl, a C.sub.1 to C.sub.7 acyl group, sulfonyl,
carbamoyl, or urea, in each instance optionally substituted with
one or more substituents, and when one or more substituents are
present, such substituents are the same or different and
independently halo, amino, monosubstituted amino, disubstituted
amino, hydroxy, or carboxy; R.sub.6a and R.sub.6b are each
independently H, alkyl, haloalkyl, cycloalkyl, alkoxy, alkythio,
halo, nitro, acyl, cyano, aryl, alkylaryl, aryloxy, oxo, amino,
monosubstituted amino, disubstituted amino, sulfonamide, hydroxy,
carboxy, or alkoxy-carbonyl; R.sub.7 is a C.sub.1 to C.sub.17
linear or branched alkyl, cycloalkyl, or alkylcycloalkyl,
optionally substituted with one or more substituents comprising
oxo, terminal amide, amino, monosubstituted amino, disubstituted
amino, or nitrile; and R.sub.8 and R.sub.9 are each independently H
or a C.sub.1 to C.sub.17 linear or branched alkyl, cycloalkyl, or
alkylcycloalkyl, optionally substituted with one or more
substituents comprising oxo, amino, monosubstituted amino,
disubstituted amino, or nitrile, provided that R.sub.8 and R.sub.9
taken together can form heterocycloakyl, optionally
substituted.
2. The compound of claim 1 in which R.sub.1 is: --CH.sub.3,
--CH.sub.2--CH.sub.3, or --CH--(CH.sub.3).sub.2.
3. The compound of claim 1 in which R.sub.5 is: --H, --CH.sub.3,
--CH.sub.2--CH.sub.3, --CH--(CH.sub.3).sub.2,
--CH.sub.2--CH--(CH.sub.3).sub.2, --C(.dbd.O)--CH.sub.3,
--C(.dbd.O)--O--CH.sub.2--CH.sub.3,
--C(.dbd.O)--NH--CH.sub.2--CH.sub.3,
--C(.dbd.O)--CH--(CH.sub.3).sub.2, or --methyl-cyclopropane.
4. The compound of claim 1 in which at least one of R.sub.6a and
R.sub.6b are halo.
5. The compound of claim 4 in which one of R.sub.6a and R.sub.6b
are halo.
6. The compound of claim 1 wherein: R.sub.1 is --R.sub.7; R.sub.2
and R.sub.3 are each independently a C.sub.1 to C.sub.17 linear
alkyl; R.sub.4 is --NR.sub.8R.sub.9; R.sub.5 is a C.sub.1 to
C.sub.17 linear or branched alkyl; R.sub.6a and R.sub.6b are each
independently H or halo; R.sub.7 is a C.sub.1 to C.sub.17 linear or
branched alkyl; and R.sub.8 and R.sub.9 are each independently H or
a C.sub.1 to C.sub.17 linear or branched alkyl.
7. The compound of claim 1 wherein: R.sub.1 is --R.sub.7; R.sub.2
and R.sub.3 are each independently a C.sub.1 to C.sub.17 linear
alkyl which taken together form cycloalkyl; R.sub.4 is
--NR.sub.8R.sub.9; R.sub.5 is a C.sub.1 to C.sub.17 linear or
branched alkyl; R.sub.6a and R.sub.6b are each independently H or
halo; R.sub.7 is a C.sub.1 to C.sub.17 linear or branched alkyl;
and R.sub.8 and R.sub.9 are each independently H or a C.sub.1 to
C.sub.17 linear or branched alkyl.
8. The compound of claim 1 which is a compound of formula II:
##STR00059##
9. The compound of claim 8 in which R.sub.1 is: --CH.sub.3,
--CH.sub.2--CH.sub.3, or --CH--(CH.sub.3).sub.2.
10. The compound of claim 8 in which R.sub.5 is: --H, --H.sub.3,
--CH.sub.2--CH.sub.3, --CH--(CH.sub.3).sub.2,
--CH.sub.2--CH--(CH.sub.3).sub.2, --C(.dbd.O)--CH.sub.3,
--C(.dbd.O)--O--CH.sub.2--CH.sub.3,
--C(.dbd.O)--NH--CH.sub.2--CH.sub.3,
--C(.dbd.O)--CH--(CH.sub.3).sub.2, or methyl-cyclopropane.
11. The compound of claim 8 in which at least one of R.sub.6a and
R.sub.6b are halo.
12. The compound of claim 1 in which one of R.sub.6a and R.sub.6b
are halo.
13. The compound of claim 8 wherein: R.sub.1 is --R.sub.7; R.sub.2
and R.sub.3 are each independently a C.sub.1 to C.sub.17 linear
alkyl; R.sub.4 is --NR.sub.8R.sub.9; R.sub.5 is a C.sub.1 to
C.sub.17 linear or branched alkyl; R.sub.6a and R.sub.6b are each
independently H or halo; R.sub.7 is a C.sub.1 to C.sub.17 linear or
branched alkyl; and R.sub.8 and R.sub.9 are each independently H or
a C.sub.1 to C.sub.17 linear or branched alkyl.
14. The compound of claim 8 wherein: R.sub.1 is --R.sub.7; R.sub.2
and R.sub.3 are each independently a C.sub.1 to C.sub.17 linear
alkyl which taken together form cycloalkyl; R.sub.4 is
--NR.sub.8R.sub.9; R.sub.5 is a C.sub.1 to C.sub.17 linear or
branched alkyl; R.sub.6a and R.sub.6b are each independently H or
halo; R.sub.7 is a C.sub.1 to C.sub.17 linear or branched alkyl;
and R.sub.8 and R.sub.9 are each independently H or a C.sub.1 to
C.sub.17 linear or branched alkyl.
15. The compound of claim 1 which is a compound of formula III:
##STR00060## or an enantiomer, stereoisomer or diastereoisomer
thereof, or a pharmaceutically acceptable salt thereof.
16. A pharmaceutical composition comprising a compound of claim 1,
or a pharmaceutically acceptable salt of claim 1, and a
pharmaceutically acceptable carrier.
17. The pharmaceutical composition of claim 16, further comprising
at least one additional active pharmaceutical agent.
18. A method of treating a patient with a disease, disorder,
condition or syndrome responsive to modulation of a melanocortin
receptor, comprising administration to the patient of a
pharmaceutically effective amount of a pharmaceutical composition
of claim 16.
19. The method of claim 18, wherein the disease, disorder,
condition or syndrome responsive to modulation of a melanocortin
receptor comprises obesity.
20. The method of claim 18, wherein the disease, disorder,
condition or syndrome responsive to modulation of a melanocortin
receptor comprises metabolic syndrome.
21. The method of claim 18, wherein the disease, disorder,
condition or syndrome responsive to modulation of a melanocortin
receptor comprises pro-opiomelanocortin deficiency due to mutations
in the POMC gene (POMC heterozygous deficiency obesity),
Prader-Willi syndrome, obesity due to MC4r deficiency, leptin
deficiency obesity, leptin receptor deficiency obesity, Bardet
Biedl syndrome or Alstrom syndrome.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation application of
International Application No. PCT/US2019/051423, published as
International Publication No. WO 2020/060983, entitled "Orally
Active Melanocortin Receptor-4 Compounds", filed on Sep. 17, 2019,
which in turn claimed priority to and the benefit of the filing of
U.S. Provisional Patent Application Ser. No. 62/732,680 entitled
"Orally Active Melanocortin Receptor-4 Compounds", filed Sep. 18,
2018, and the specification and claims thereof of each are
incorporated herein by reference.
BACKGROUND OF THE INVENTION
Field of the Invention (Technical Field)
[0002] This invention provides substituted piperazine compounds
that are agonists or partial agonists at melanocortin receptor-4
and use thereof in the treatment of a wide variety of diseases,
syndromes and conditions, including in the treatment of obesity,
diabetes, metabolic syndrome and related indications, diseases or
disorders.
Background
[0003] A family of melanocortin receptor types and subtypes have
been identified, including melanocortin receptor-1 (MC1r) expressed
on normal human melanocytes, melanoma cells and other cells;
melanocortin receptor-2 (MC2r) for ACTH (adrenocorticotropin),
expressed in cells of the adrenal gland; melanocortin receptor-3
and melanocortin receptor-4 (MC3r and MC4r), expressed in cells in
the hypothalamus, mid-brain, brainstem and in peripheral tissues;
and melanocortin receptor-5 (MC5r), expressed in a wide
distribution of peripheral tissues. MC1r is believed to be
associated with hair and skin pigmentation, mediation of
inflammation and other functions; MC2r is believed to mediate
steroidogenesis; MC3r is believed to be associated with energy
homeostasis, food intake, and inflammation; MC4r is believed to be
associated with feeding behavior, energy homeostasis, and sexual
functioning; and MC5r is believed to be involved in exocrine gland
system regulation and other functions.
[0004] Significant work has been done in determining the structure
of melanocortin receptors, including both the nucleic acid
sequences encoding for the receptors and the amino acid sequences
constituting the receptors. MC4r is a G protein-coupled,
7-transmembrane receptor that is expressed in the brain,
particularly the paraventricular nucleus of the hypothalamus and
the dorsal motor nucleus of the vagus nerve within the hindbrain,
but is also expressed peripherally, including intestinal cells.
[0005] MC4r inactivation has been shown to result in obesity
(Hadley, Ann N Y Acad Sci, 885:1-21 (1999)), with activation of
MC4r, through the endogenous agonist .alpha.-melanocyte stimulating
hormone (.alpha.-MSH) or synthetic analogs, resulting in weight
loss (Hsiung et al., Endocrinology, 146:5257-5266 (2005)).
.alpha.-MSH is believed to be the principle endogenous MC4r
agonist.
[0006] A number of small molecule compounds asserted to be agonists
or partial agonists at one or more melanocortin receptors have been
developed, including the compounds disclosed in EP3150578A1 (filed
28 May 2015, claiming priority to international application
PCT/JP2015/065469), WO2010/065801A1 (PCT/US2009/066671, filed 4
Dec. 2009), WO2010/065802 A2 (PCT/US2009/066672, filed 4 Dec.
2009), WO2010/065800 A1 (PCT/US2009/066669, filed 4 Dec. 2009),
WO2010/065799 A2 (PCT/US2009/066668, filed 4 Dec. 2009), and U.S.
Pat. Nos. 9,493,456, 7,732,451, 7,727,991, 7,727,990, 7,718,802,
and 7,354,923, among others. However, there are no approved small
molecule melanocortin agonist or partial agonist products in the
United States.
[0007] There is a significant need for compounds specific for MC4r
for attenuating food intake and body weight gain, and for treatment
of obesity, diabetes mellitus type 2, metabolic syndrome and
related conditions and indications, as well as compounds specific
for MC4r for treatment of conditions relating to various expression
or receptor genetic diseases such as pro-opiomelanocortin
deficiency due to mutations in the POMC gene (POMC heterozygous
deficiency obesity), Prader-Willi syndrome, obesity due to MC4r
deficiency, leptin receptor deficiency obesity, Bardet Biedl
syndrome, Alstrom syndrome, and various other diseases, conditions,
genetic deficiencies, metabolic disorders, and syndromes. It is
against this background that the present invention was made.
SUMMARY OF THE INVENTION (DISCLOSURE OF THE INVENTION)
[0008] In one aspect, the invention provides compounds of formula
I:
##STR00002##
or an enantiomer, stereoisomer or diastereoisomer thereof, or a
pharmaceutically acceptable salt thereof, wherein: [0009] R.sub.1
is --R.sub.7 or --NR.sub.8R.sub.9; [0010] R.sub.2 and R.sub.3 are
each independently a C.sub.1 to C.sub.17 linear or branched alkyl,
cycloalkyl, alkylcycloalkyl, aryl, and alkylaryl, optionally
substituted with one or more substituents, and when one or more
substituents are present, such substituents are the same or
different and independently comprising oxo, carboxy, amino,
monosubstituted amino, disubstituted amino, or nitro, and when
R.sub.2 and R.sub.3 are different, including all sterioisomers
thereof, provided that taken together R.sub.2 and R.sub.3 can form
cycloalkyl, optionally substituted, or alternatively that taken
together one of R.sub.2 and R.sub.3 and one of R.sub.8 and R.sub.9
comprising a part of R.sub.4 can form heterocycloalkyl, optionally
substituted; [0011] R.sub.4 is --OH, --OR.sub.7, or
--NR.sub.8R.sub.9; [0012] R.sub.5 is [0013] H, [0014] a C.sub.1 to
C.sub.17 linear or branched alkyl, cycloalkyl, or alkylcycloalkyl,
[0015] a C.sub.1 to C.sub.7 acyl group, [0016] sulfonyl, [0017]
carbamoyl or [0018] urea, [0019] in each instance optionally
substituted with one or more substituents, and when one or more
substituents are present, such substituents are the same or
different and independently halo, amino, monosubstituted amino,
disubstituted amino, hydroxy, or carboxy; [0020] R.sub.6a and
R.sub.6b are each independently H, alkyl, haloalkyl, cycloalkyl,
alkoxy, alkythio, halo, nitro, acyl, cyano, aryl, alkylaryl,
aryloxy, oxo, amino, monosubstituted amino, disubstituted amino,
sulfonamide, hydroxy, carboxy, or alkoxy-carbonyl; [0021] R.sub.7
is a C.sub.1 to C.sub.17 linear or branched alkyl, cycloalkyl, or
alkylcycloalkyl, optionally substituted with one or more
substituents comprising oxo, terminal amide, amino, monosubstituted
amino, disubstituted amino, or nitrile; and [0022] R.sub.8 and
R.sub.9 are each independently H or a C.sub.1 to C.sub.17 linear or
branched alkyl, cycloalkyl, or alkylcycloalkyl, optionally
substituted with one or more substituents comprising oxo, amino,
monosubstituted amino, disubstituted amino, or nitrile, provided
that R.sub.8 and R.sub.9 taken together can form heterocycloakyl,
optionally substituted.
[0023] In one aspect, in the compound of formula I the group
R.sub.1 is --CH.sub.3, --CH.sub.2--CH.sub.3, or
--CH--(CH.sub.3).sub.2. In another aspect, in the compound of
formula I the group R.sub.5 is --H, --CH.sub.3,
--CH.sub.2--CH.sub.3, --CH--(CH.sub.3).sub.2,
--CH.sub.2--CH--(CH.sub.3).sub.2, --C(.dbd.O)--CH.sub.3,
--C(.dbd.O)--O--CH.sub.2--CH.sub.3,
--C(.dbd.O)--NH--CH.sub.2--CH.sub.3,
--C(.dbd.O)--CH--(CH.sub.3).sub.2, or -methyl-cyclopropane. In yet
another aspect, in the compound of formula I at least one of
R.sub.6a and R.sub.6b are halo, or alternatively only one of
R.sub.6a and R.sub.6b are halo.
[0024] In another aspect, the invention provides a compound of
formula I wherein: [0025] R.sub.1 is --R.sub.7; [0026] R.sub.2 and
R.sub.3 are each independently a C.sub.1 to C.sub.17 linear alkyl;
[0027] R.sub.4 is --NR.sub.8R.sub.9; [0028] R.sub.5 is a C.sub.1 to
C.sub.17 linear or branched alkyl; [0029] R.sub.6a and R.sub.6b are
each independently H or halo; [0030] R.sub.7 is a C.sub.1 to
C.sub.17 linear or branched alkyl; and [0031] R.sub.8 and R.sub.9
are each independently H or a C.sub.1 to C.sub.17 linear or
branched alkyl.
[0032] In yet another aspect, the invention provides a compound of
formula I wherein: [0033] R.sub.1 is --R.sub.7; [0034] R.sub.2 and
R.sub.3 are each independently a C.sub.1 to C.sub.17 linear alkyl
which taken together form cycloalkyl; [0035] R.sub.4 is
--NR.sub.8R.sub.9; [0036] R.sub.5 is a C.sub.1 to C.sub.17 linear
or branched alkyl; [0037] R.sub.6a and R.sub.6b are each
independently H or halo; [0038] R.sub.7 is a C.sub.1 to C.sub.17
linear or branched alkyl; and [0039] R.sub.8 and R.sub.9 are each
independently H or a C.sub.1 to C.sub.17 linear or branched
alkyl.
[0040] In another aspect, the invention accordingly provides a
compound of formula II:
##STR00003##
wherein the variables R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5,
R.sub.6a, and R.sub.6b are as defined for formula I.
[0041] In another aspect, in the compound of formula II the group
R.sub.1 is --CH.sub.3, --CH.sub.2--CH.sub.3, or
--CH--(CH.sub.3).sub.2. In yet another aspect, in the compound of
formula II the group R.sub.5 is --H, --CH.sub.3,
--CH.sub.2--CH.sub.3, --CH--(CH.sub.3).sub.2,
--CH.sub.2--CH--(CH.sub.3).sub.2, --C(.dbd.O)--CH.sub.3,
--C(.dbd.O)--O--CH.sub.2--CH.sub.3,
--C(.dbd.O)--NH--CH.sub.2--CH.sub.3,
--C(.dbd.O)--CH--(CH.sub.3).sub.2, or -methyl-cyclopropane. In yet
another aspect, in the compound of formula II at least one of
R.sub.6a and R.sub.6b are halo, or alternatively only one of
R.sub.6a and R.sub.6b are halo.
[0042] In another aspect, the invention provides a compound of
formula II wherein: [0043] R.sub.1 is --R.sub.7; [0044] R.sub.2 and
R.sub.3 are each independently a C.sub.1 to C.sub.17 linear alkyl;
[0045] R.sub.4 is --NR.sub.8R.sub.9; [0046] R.sub.5 is a C.sub.1 to
C.sub.17 linear or branched alkyl; [0047] R.sub.6a and R.sub.6b are
each independently H or halo; [0048] R.sub.7 is a C.sub.1 to
C.sub.17 linear or branched alkyl; and [0049] R.sub.8 and R.sub.9
are each independently H or a C.sub.1 to C.sub.17 linear or
branched alkyl.
[0050] In yet another aspect, the invention provides a compound of
formula I wherein: [0051] R.sub.1 is --R.sub.7; [0052] R.sub.2 and
R.sub.3 are each independently a C.sub.1 to C.sub.17 linear alkyl
which taken together form cycloalkyl; [0053] R.sub.4 is
--NR.sub.8R.sub.9; [0054] R.sub.5 is a C.sub.1 to C.sub.17 linear
or branched alkyl; [0055] R.sub.6a and R.sub.6b are each
independently H or halo; [0056] R.sub.7 is a C.sub.1 to C.sub.17
linear or branched alkyl; and [0057] R.sub.8 and R.sub.9 are each
independently H or a C.sub.1 to C.sub.17 linear or branched alkyl.
In yet another aspect, the invention provides a compound of formula
III:
##STR00004##
[0057] Including an enantiomer, stereoisomer or diastereoisomer
thereof, or a pharmaceutically acceptable salt thereof.
[0058] The invention further includes a pharmaceutical composition
including a compound of any of formulas I, II or III as provided
above, or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier. The pharmaceutical composition
may further include at least one additional active pharmaceutical
agent.
[0059] The invention further provides a method of treating a
patient with a disease, disorder, condition or syndrome responsive
to modulation of a melanocortin receptor, comprising administration
to the patient of a pharmaceutically effective amount of the
pharmaceutical composition. The disease, disorder, condition or
syndrome responsive to modulation of a melanocortin receptor may
include obesity or may include metabolic syndrome. In another
aspect, the the disease, disorder, condition or syndrome responsive
to modulation of a melanocortin receptor may include any one or
more of pro-opiomelanocortin deficiency due to mutations in the
POMC gene (POMC heterozygous deficiency obesity), Prader-Willi
syndrome, obesity due to MC4r deficiency, leptin receptor
deficiency obesity, Bardet Biedl syndrome or Alstrom syndrome.
[0060] The invention further comprises a method for inhibiting food
uptake in a mammal, comprising administering a pharmaceutically
sufficient amount of a composition comprising a compound of this
invention or pharmaceutically acceptable salt thereof, and
particularly an MC4r selective agonist or partial agonist. The
composition may further comprise a pharmaceutically acceptable
carrier. In the method, administering may include administering by
any method of administration, including but not limited to oral or
systemic administration.
[0061] One object of this invention is to provide a melanocortin
receptor-specific pharmaceutical for use in treatment of eating
disorders.
[0062] Another object of this invention is to provide an oral
pharmaceutical for use in treatment of melanocortin
receptor-specific, and preferably MC4r-specific, mediated
disorders, diseases, conditions and syndromes.
[0063] Another object is to provide a melanocortin
receptor-specific pharmaceutical which is effective by oral
administration.
[0064] Another object is to provide an oral administration
melanocortin receptor-specific pharmaceutical with minimal or no
effects on increasing blood pressure and/or decreasing heart
rate.
[0065] Other objects, advantages and novel features, and the
further scope of applicability of this invention, will be set forth
in part in the detailed description to follow, and in part will
become apparent to those skilled in the art upon examination of the
following, or may be learned by practice of this invention. The
objects and advantages of this invention may be realized and
attained by means of the instrumentalities and combinations
particularly pointed out in the appended claims.
BRIEF DESCRIPTION OF THE DRAWINGS
[0066] The accompanying drawings, which are incorporated into and
form a part of the specification, illustrate one or more
embodiments of the present invention and, together with the
description, serve to explain the principles of the invention. The
drawings are only for the purpose of illustrating one or more
preferred embodiments of the invention and are not to be construed
as limiting the invention.
[0067] FIG. 1 is a plot of mean plasma concentration (ng/mL) over
24 hours of Compound 7.1 in Sprague-Dawley rats following oral
administration of Compound 1 at a dose of 30 mg/kg.
[0068] FIG. 2 is a bar graph of mean plasma concentration (ng/mL)
of Compound 7.1 in DIO mice at one hour following oral
administration of Compound 1 at doses of 3 mg/kg, 10 mg/kg and 30
mg/kg.
[0069] FIG. 3 is a plot of decrease in body weight percent change
in leptin-deficient male mice receiving 30 mg/kg of Compound 7.1
compared to leptin-deficient male mice receiving vehicle
(normalized to vehicle).
[0070] FIG. 4 is a plot of body weight (BW) percent change from
vehicle in MC4-/-(MC4r knockout mice) compared to DIO C57Bl/6 mice
over time (normalized to vehicle).
DESCRIPTION OF THE PREFERRED EMBODIMENTS
Best Modes for Carrying Out the Invention
1. Definitions
[0071] As used herein, the term "alkyl" or "alkyl group" means a
saturated unbranched or branched hydrocarbon chain.
(C.sub.1-C.sub.6) alkyl means an alkyl having from 1 to 6 carbon
atoms. Non-limiting examples of (C.sub.1-C.sub.6) alkyl groups
include methyl, ethyl, propyl, isopropyl, 2-methyl-1-propyl,
2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl,
2-methyl-3-butyl, 2,2-dimethyl-1-propyl, 2-methyl-1-pentyl,
3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl,
3-methyl-2-pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl,
3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl, isobutyl, t-butyl,
pentyl, isopentyl, neopentyl, and hexyl. Alkyl includes longer
alkyl groups, such as heptyl and octyl. An alkyl group can be
unsubstituted or optionally substituted with one or two suitable
substituents.
[0072] As used herein, the term "alkenyl" or "alkenyl group" means
an unbranched or branched hydrocarbon chain having one or more
double bonds therein (i.e., comprising an alkene or olefin). The
double bond of an alkenyl group can be unconjugated or conjugated
to another unsaturated group. Suitable alkenyl groups include, but
are not limited to (C.sub.2-C.sub.6) alkenyl groups, such as vinyl,
allyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl,
hexadienyl, 2-ethylhexenyl, 2-propyl-2-butenyl,
4-(2-methyl-3-butene)-pentenyl. An alkenyl group can be
unsubstituted or optionally substituted with one or two suitable
substituents.
[0073] As used herein, the term "alkynyl" or "alkynyl group" means
an unbranched or branched hydrocarbon chain having one or more
triple bonds therein. The triple bond of an alkynyl group can be
unconjugated or conjugated to another unsaturated group. Suitable
alkynyl groups include, but are not limited to, (C.sub.2-C.sub.6)
alkynyl groups, such as ethynyl, propynyl, butynyl, pentynyl,
hexynyl, methylpropynyl, 4-methyl-1-butynyl, 4-propyl-2-pentynyl,
and 4-butyl-2-hexynyl. An alkynyl group can be unsubstituted or
optionally substituted with one or two suitable substituents.
[0074] As used herein, the term "aralkyl" means a radical
--R.sup.aR.sup.b where R.sup.a is an alkylene (a bivalent alkyl)
group and R.sup.b is an aryl group as defined herein. Examples of
aralkyl groups include benzyl, phenylethyl,
3-(3-chlorophenyl)-2-methylpentyl, and the like.
[0075] As used herein, the term "aryl" or "aryl group" means a
monocyclic or polycyclic (e.g., bicyclic) aromatic ring system
comprising carbon and hydrogen atoms. The term "aryl" also includes
polycyclic aromatic ring systems wherein at least one ring is
aromatic and one or more rings are non-aromatic (including
saturated or partially saturated rings). Non-limited examples
include phenyl, tolyl, anthacenyl, fluorenyl, indenyl, azulenyl,
naphthyl, 1-naphthyl, 2-naphthyl, and biphenyl as well as
benzo-fused carbocyclic moieties such as
5,6,7,8-tetrahydronaphthyl. An aryl group can be unsubstituted or
optionally substituted with one or more suitable substituents as
defined below. An aryl group may be fused to a cycloalkyl group,
fused to another aryl group, fused to a heteroaryl group, or fused
to a heterocycloalkyl group. Preferred aryl groups include, but are
not limited to, monocyclic or bicyclic aromatic hydrocarbon
radicals of 6 to 12 ring atoms, and optionally substituted
independently with one or more substituents selected from alkyl,
haloalkyl, cycloalkyl, alkoxy, alkythio, halo, nitro, acyl, cyano,
aryl, alkylaryl, aryloxy, amino, monosubstituted amino,
disubstituted amino, sulfonamide, hydroxy, carboxy, or
alkoxy-carbonyl.
[0076] As used herein, the term "heteroaryl" or "heteroaryl group"
means a monocyclic or polycyclic aromatic ring comprising carbon
atoms, hydrogen atoms, and one or more heteroatoms, preferably 1 to
4 heteroatoms, independently selected from nitrogen, oxygen, and
sulfur. Non-limiting examples of heteroaryl groups include pyridyl,
pyridazinyl, pyrazyl, indolyl, triazinyl, pyrrolyl, pyrazolyl,
imidazolyl, (1,2,3,)-triazolyl, (1,2,4)-triazolyl, pyrazinyl,
pyrimidinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl,
thiadiazolyl, furyl, phienyl, isoxazolyl, oxazolyl, pyrazolyl,
tetrazolyl, triazolyl, oxadiazolyl, thiadiazolyl, isoxazolyl,
triazinyl, and pyrazinyl. Bicyclic heteroaromatic rings include,
but are not limited to, benzothiadiazolyl, indolyl,
benzothiophenyl, benzofuryl, benzimidazolyl, benzisoxazolyl,
benzothiazolyl, quinolinyl, benzotriazolyl, benzoxazolyl,
isoquinolinyl, purinyl, furopyridinyl and thienopyridinyl. A
heteroaryl can be unsubstituted or optionally substituted with one
or more suitable substituents as defined below. A heteroaryl group
optionally may be fused to another heteroaryl group, fused to an
aryl group, fused to a cycloalkyl group, or fused to a
heterocycloalkyl group.
[0077] As used herein, the term "cycloalkyl" or "cycloalkyl group"
means a monocyclic or polycyclic saturated ring comprising carbon
and hydrogen atoms and having no carbon-carbon multiple bonds.
Examples of cycloalkyl groups include, but are not limited to,
(C.sub.3-C.sub.7) cycloalkyl groups, such as cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl, and saturated
cyclic and bicyclic terpenes. A cycloalkyl group can be
unsubstituted or optionally substituted with one or more suitable
substituents as defined below. A cycloalkyl group optionally may be
fused to another cycloalkyl group, fused to an aryl group, fused to
a heteroaryl group, or fused to a heterocycloalkyl group.
[0078] As used herein, the term "heterocycloalkyl" or
"heterocycloalkyl group" means a monocyclic or polycyclic ring
comprising carbon and hydrogen atoms and at least one heteroatom,
preferably, 1 to 3 heteroatoms selected from nitrogen, oxygen, and
sulfur. A heterocycloalkyl group may be fused to an aryl or
heteroaryl group. Examples of heterocycloalkyl groups include, but
are not limited to, pyrrolidinyl, pyrrolidino, piperidinyl,
piperidino, piperazinyl, piperazino, morpholinyl, morpholino,
thiomorpholinyl, thiomorpholino, and pyranyl. A heterocycloalkyl
group can be unsubstituted or optionally substituted with one or
more suitable substituents as defined below. A heterocycloalkyl
group optionally may be fused to a cycloalkyl group, fused to an
aryl group, fused to a heteroaryl group, or fused to another
heterocycloalkyl group. For example, a heterocycloalkyl group can
be fused to or substituted with an aryl group or heteroaryl group,
for example, but not limited to, 1,2,3,4-tetrahydroisoquinolinyl
and 1,2,3,4-tetrahydroquinolinyl, tetrahydronaphthyridinyl,
phenylpiperidinyl, and piperidinylpyridinyl. A heterocycloalkyl
group may be a monocyclic or bicyclic ring, such as a monocyclic
ring comprising from 3 to 6 carbon atoms and from 1 to 3
heteroatoms, referred to herein as (C.sub.3-C.sub.6)
heterocycloalkyl. A heterocycloalkyl group may also be fused to or
substituted with an aryl group or a heteroaryl group.
[0079] As used herein, the term "alkoxy" means an --O-alkyl group,
wherein alkyl is as defined above. An alkoxy group can be
unsubstituted or optionally substituted with one or two suitable
substituents. Preferably, the alkyl chain of an alkyloxy group is
from 1 to 6 carbon atoms in length, referred to herein as
"(C.sub.1-C.sub.6) alkoxy".
[0080] As used herein, the term "aryloxy" means an --O-aryl group,
wherein aryl is as defined above. An aryloxy group can be
unsubstituted or optionally substituted with one or two suitable
substituents. Preferably, the aryl ring of an aryloxy group is a
monocyclic ring, wherein the ring comprises 6 carbon atoms,
referred to herein as "(C.sub.6)aryloxy".
[0081] As used herein, the term "carbamoyl" group means the radical
--C(.dbd.O)N(R').sub.2, wherein R' is chosen from the group
consisting of hydrogen, alkyl, and aryl.
[0082] As used herein, a "carbonyl" group means a divalent group of
the formula C(.dbd.O).
[0083] As used herein, an "oxo" group means a group of the formula
(.dbd.O).
[0084] As used herein, the term "acyl" means a group
R--C(.dbd.O)--, where R is an organic group, including but not
limited to a C.sub.1 to C.sub.7 alkyl. An example is the acetyl
group CH.sub.3--C(.dbd.O)--, referred to herein sometimes as "Ac".
A moiety is "acylated" when an aryl, alkyl or substituted alkyl
group as defined above is bonded through one or more carbonyl
{--(C.dbd.O)--} groups.
[0085] As used herein, an "amide" means compounds that have a
trivalent nitrogen attached to a carbonyl group
(--C(.dbd.O)--NH.sub.2), such as for example methylamide,
ethylamide, propylamide, and the like.
[0086] As used herein, an "amine" means compounds that contain an
amino group (--NH.sub.2), monosubstituted amino group (--NHR) or
disubstituted amino group (--NRR), where each R is independently a
suitable substituent.
[0087] As used herein a "nitrile" means compounds that are
carboxylic acid derivatives and contain a (--C.ident.N) group bound
to an organic group.
[0088] As used herein, the term "halogen" means fluorine, chlorine,
bromine, or iodine. Correspondingly, the meaning of the terms
"halo" and "Hal" encompass fluoro, chloro, bromo, and iodo.
[0089] As used herein, the term "sulfonamide" means compounds of
the formula --R--S(.dbd.O).sub.2--NH.sub.2, where R any organic
group, including but not limited to alkyl, and where sulfonamide is
a suitable substituent, where R is a portion of the substituted
group.
[0090] As used herein, the term "sulfonyl" means compounds of the
formula --R--S(.dbd.O).sub.2--R', and where sulfonyl is a suitable
substituent, where R is a portion of the substituted group and R'
is hydrogen or an organic group including but not limited to alkyl,
O-alkyl, aryl, alkenyl or aralkyl.
[0091] As used herein, the term "urea" means compounds of the
formula formula --R--C(.dbd.O)--O--R', and where urea is a suitable
substituent, where R is a portion of the substituted group and
where R' is hydrogen or an organic group including but not limited
to alkyl, aryl, alkenyl or aralkyl.
[0092] As used herein, the abbreviation ("PG") or term "protecting
group" means a nitrogen protecting group that replaces an amino
hydrogen for the purpose of protecting against side reactions and
degradation during a reaction sequence. Nitrogen protecting groups
useful in the invention include nitrogen protecting groups well
known in the synthetic arts, including, but not limited to, Boc,
Fmoc, 2-chlorobenzyloxycarbonyl, alloc, benzyloxycarbonyl (Z),
2-(4-biphenylyl)propyl-2-oxycarbonyl (Bpoc),
1-adamantyloxycarbonyl, triphenylmethyl (trityl), and toluene
sulphonyl.
[0093] As used herein, the term "suitable substituent" means a
group that does not nullify the synthetic, therapeutic or
pharmaceutical utility of the compounds of the invention or the
intermediates useful for preparing them. Examples of suitable
substituents include, but are not limited to: alkyl; haloalkyl;
[0094] cycloalkyl; alkoxy; alkythio; halo; nitro; acyl; cyano;
aryl; alkylaryl; aryloxy; amino; monosubstituted amino;
disubstituted amino; carbamoyl, urea; sulfonamide; sulfonyl; oxo,
hydroxyl; carboxy; alkoxy-carbonyl; alkenyl; alkynyl; heteroaryl;
heterocycloalkyl; O-alkenyl; O-alkynyl; oxo; CF.sub.3; NO.sub.2;
NH.sub.2; NH(alkyl); N(alkyl).sub.2; NH(aryl); N(aryl).sub.2;
C(.dbd.O)NH.sub.2; C(.dbd.O)NH(alkyl); C(.dbd.O)N(alkyl).sub.2;
C(.dbd.O)NH(aryl); C(.dbd.O)N(aryl).sub.2; OC(.dbd.O)NH.sub.2;
C(.dbd.O)NH(heteroaryl); C(.dbd.O)N(heteroaryl).sub.2;
C(.dbd.O)NH(aralkyl); C(.dbd.O)N(aralkyl).sub.2;
OC(.dbd.O)NH(alkyl); OC(.dbd.O)N(alkyl).sub.2; OC(.dbd.O)NH(aryl);
OC(.dbd.O)N(aryl).sub.2; OC(.dbd.O)NH(aralkyl);
OC(.dbd.O)N(aralkyl).sub.2; C(.dbd.O)(alkyl); C(.dbd.O)(aryl);
C(.dbd.O)(aralkyl); C(.dbd.O)O(alkyl); C(.dbd.O)O(aryl);
C(.dbd.O)O(aralkyl); OC(.dbd.O)(alkyl); OC(.dbd.O)(aryl);
OC(.dbd.O)(aralkyl); OC(.dbd.O)O(alkyl); OC(.dbd.O)O(aryl);
OC(.dbd.O)O(aralkyl); S-alkyl; S-alkenyl; S-alkynyl; S-aryl,
S(.dbd.O).sub.2-alkyl; S(.dbd.O).sub.2--O-alkyl;
S(.dbd.O).sub.2-alkenyl; S(.dbd.O).sub.2-aralkyl; and
S(.dbd.O).sub.2-aryl. One of skill in art can readily choose a
suitable substituent based on the synthesis, stability and
pharmacological activity of the compound of the invention.
[0095] The term "composition", as in pharmaceutical composition, is
intended to encompass a product comprising the active
ingredient(s), and the inert ingredient(s) that make up the
carrier, as well as any product which results, directly or
indirectly, from combination, complexation or aggregation of any
two or more of the ingredients, or from dissociation of one or more
of the ingredients, or from other types of reactions or
interactions of one or more of the ingredients. Accordingly, the
pharmaceutical compositions of this invention encompass any
composition made by admixing a compound of this invention and a
pharmaceutically acceptable carrier.
[0096] The term "EC.sub.50" is intended to include the molar
concentration of an agonist, including a partial agonist, which
produced 50% of the maximum possible response for that agonist or
partial agonist. By way of example, a compound which, at a
concentration of 72 nM, produces 50% of the maximum possible
response for that compound as determined in a cAMP assay, has an
EC.sub.50 of 72 nM. Unless otherwise specified, the molar
concentration associated with an EC.sub.50 determination is in
nanomoles (nM).
[0097] The term "Ki (nM)" is intended to include the equilibrium
receptor binding affinity representing the molar concentration of a
competing compound that binds to half the binding sites of a
receptor at equilibrium in the absence of a competitor. In general,
the Ki is inversely correlated to the affinity of the compound for
the receptor, such that if the Ki is low, the affinity is high. Ki
may be determined using the equation of Cheng and Prusoff (Cheng
Y., Prusoff W. H., Biochem. Pharmacol. 22: 3099-3108, 1973):
Ki = IC 50 1 + [ ligand ] K d ##EQU00001##
where "ligand" is the concentration of ligand, which may be a
radioligand, and K.sub.d is an inverse measure of receptor affinity
which produces 50% receptor occupancy. Unless otherwise specified,
the molar concentration associated with a Ki determination is nM.
IC.sub.50 is the concentration of ligand at which 50% of
receptor-bound radioligand or other ligand is displaced from the
receptor sites in a competitive receptor binding displacement
assay.
[0098] By "relative efficacy" is meant the maximal functional
activity achievable by a compound in a specified melanocortin
receptor expressing cell system, such as the maximal stimulation of
adenylyl cyclase. The maximal stimulation achieved by .alpha.-MSH
or NDP-.alpha.-MSH is designated as an intrinsic activity of 1.0
(or 100%) and a compound capable of stimulating half the maximal
activity that of .alpha.-MSH or NDP-.alpha.-MSH is designated as
having an intrinsic activity of 0.5 (or 50%). A compound of this
invention that under assay conditions described herein has an
intrinsic activity of 0.7 (70%) or higher is classified as an
agonist, a compound with intrinsic activity between 0.1 (10%) and
0.7 (70%) is classified as a partial agonist, and a compound with
intrinsic activity below 0.1 (10%) is classified as inactive or
having no intrinsic activity. In one aspect, the cyclic peptides of
the present invention may generally be characterized as a partial
agonist at MC4r with respect to .alpha.-MSH or NDP-.alpha.-MSH in a
human MC4r expression system approximating maximal physiologic
receptor densities.
[0099] The terms "treat," "treating" and "treatment," as used
herein, contemplate an action that occurs while a patient is
suffering from the specified disease or disorder, which reduces the
severity of the disease or disorder.
[0100] As used herein, the term "pharmaceutically effective amount"
means the amount of a compound of the invention that will elicit a
biological or medical response in the mammal that is being treated
by a medical doctor or other clinician.
[0101] As used herein, the term "prophylactically effective" or
"preventive" means the amount of a compound of the invention that
will prevent or inhibit affliction or mitigate affliction of a
mammal with a medical condition that a medical doctor or other
clinician is trying to prevent, inhibit, or mitigate before a
patient begins to suffer from the specified disease or
disorder.
[0102] The term "pharmaceutically acceptable salt(s)", as used
herein includes salts prepared from pharmaceutically acceptable
non-toxic inorganic or organic bases or acids, thereby constituting
pharmaceutically acceptable acid and base addition salts (see
Handbook of Pharmaceutical Salts: Properties, Selection and Use, P.
H. Stahl, P. G. Wermuth, IUPAC, Wiley-VCH, 2002). Acid addition
salts are formed from inorganic or organic acids. Examples of
suitable non-toxic acid addition salts are acetate, adipate,
aspartate, benzoate, besylate, bicarbonate/carbonate,
bisulphate/sulphate, borate, camsylate, citrate, cyclamate,
edisylate, esylate, formate, fumarate, gluceptate, gluconate,
glucuronate, hexafluorophosphate, hibenzate,
hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide,
isethionate, lactate, malate, maleate, malonate, mesylate,
methylsulphate, naphthylate, 2-napsylate, nicotinate, nitrate,
orotate, oxalate, palmitate, pamoate, phosphate/hydrogen
phosphate/dihydrogen phosphate, pyroglutamate, saccharate,
stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate
and xinofoate salts. Hemisalts of the acids may also be formed, for
example, hemisulphate. Base-addition salts are formed from
inorganic or organic bases. Examples of suitable non-toxic
base-addition salts are salts derived from aluminum, ammonium,
calcium, copper, ferric, ferrous, lithium, magnesium, manganic
salts, manganous, potassium, sodium, zinc, primary, secondary, and
tertiary amines, substituted amines including naturally occurring
substituted amines, cyclic amines, and basic ion exchange resins,
such as arginine, betaine, caffeine, choline,
N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol,
2-dimethylaminoethanol, ethanolamine, ethylenediamine,
N-ethyl-morpholine, N-ethylpiperidine, glucamine, glucosamine,
histidine, hydrabamine, isopropylamine, lysine, methylglucamine,
morpholine, piperazine, piperidine, polyamine resins, procaine,
purines, theobromine, TEA, trimethylamine, tripropylamine, and
tromethamine.
[0103] The chemical naming protocol and structure diagrams used
herein employ and rely on the chemical naming features as utilized
by the BIOVIA Draw 2018 program (Dassault Systemes), but other
comparable programs, such as ChemDraw programs (available from
Perkin Elmer), may similarly be employed. In general, structure
diagrams do not depict hydrogen atoms other than on heteroatoms, in
terminal groups and other special circumstances.
2. Isomeric Purity, Prodrugs and Isotopically-Substituted
Compounds
[0104] Isomeric Purity and Isolation. The compounds of the
invention can contain one or more chiral centers and/or double
bonds and, therefore, exist as stereoisomers, such as double-bond
isomers (i.e., cis-trans isomers or geometric isomers), enantiomers
(optical isomers), or diastereomers. According to the invention,
the chemical structures depicted herein, and therefore the
compounds of the invention, encompass the racemic form of compounds
as well as all enantiomers and stereoisomers, that is, both the
stereomerically pure form (e.g., geometrically pure,
enantiomerically pure, or diastereomerically pure) and enantiomeric
and stereoisomeric mixtures.
[0105] A compound is considered optically active or
enantiomerically pure (i.e., substantially the R-form or
substantially the S-form) with respect to a chiral center when the
compound is about 90% ee (enantiomeric excess) or greater,
preferably, equal to or greater than 95% ee with respect to a
particular chiral center. A compound of the invention is considered
to be in enantiomerically enriched form when the compound has an
enantiomeric excess of greater than about 80% ee, preferably
greater than about 85% ee. As used herein, a racemic mixture means
about 50% of one enantiomer and about 50% of its corresponding
enantiomer relative to all chiral centers in the molecule. Thus,
the invention encompasses all enantiomerically pure,
enantiomerically enriched, and racemic mixtures of compounds of the
invention.
[0106] Enantiomeric and stereoisomeric mixtures can be resolved
into their component enantiomers or stereoisomers by well known
methods, such as chiral-phase gas chromatography, chiral-phase high
performance liquid chromatography, crystallizing the compound as a
chiral salt complex, or crystallizing the compound in a chiral
solvent. Enantiomers and stereoisomers can also be obtained from
stereomerically- or enantiomerically-pure intermediates, reagents,
and catalysts by well known asymmetric synthetic methods.
[0107] The compounds of the invention also include, where possible,
all tautomeric isomers thereof, such as prototropic tautomerism
(e.g. hydroxypyridine--pyridone, ketone--enol, amide--imidic acid,
amine--imine), annular tautomerism, ring-chain tautomerism and
valence tautomerism.
[0108] Prodrugs. The invention is further intended to include
prodrugs of the compounds of the invention, which on administration
undergo chemical conversion by metabolic processes before becoming
active pharmacological compounds. In general, such prodrugs will be
functional derivatives of compounds of the invention, which are
readily convertible in vivo into a compound of formula (I).
[0109] Prodrugs are any covalently bonded compounds, which release
the active parent compound drug of formula (I) in vivo.
Conventional procedures for the selection and preparation of
suitable prodrug derivatives are described, for example, in Design
of Prodrugs ed. H. Bundgaard, Elsevier, 1985 and Rautio, J., et
al., "Prodrugs: design and clinical applications," Nat. Rev. Drug
Disc. 7:255-270 (2008). Typical examples of prodrugs have
biologically labile protecting groups on a functional moiety, such
as for example by esterification of hydroxyl, carboxyl or amino
functions. Broadly speaking, prodrugs include compounds that can be
oxidized, reduced, aminated, deaminated, hydroxylated,
dehydroxylated, hydrolyzed, dehydrolyzed, alkylated, dealkylated,
acylated, deacylated, phosphorylated or dephosphorylated to produce
an active parent drug of formula (I) in vivo.
[0110] Isotopically-Substituted Compounds. The subject invention
also includes compounds which are identical to those recited in
formula (I), but for the fact that one or more atoms depicted in
formula (I) are replaced by an atom having an atomic mass or mass
number different from the atomic mass or mass number usually found
in nature. Examples of isotopes that can be incorporated into
compounds of the invention include isotopes of hydrogen, carbon,
nitrogen, oxygen and fluorine, such as .sup.2H, .sup.3H, .sup.11C,
.sup.13C, .sup.14C, .sup.15N, .sup.18O, .sup.17O or .sup.19F.
Compounds of this invention and pharmaceutically acceptable salts
or solvates of said compounds which contain the aforementioned
isotopes and/or other isotopes of other atoms are within the scope
of this invention. Certain isotopically-labeled compounds of this
invention, for example those into which radioactive isotopes such
as .sup.3H and .sup.14C are incorporated, may have use in a variety
of assays, such as drug and/or substrate tissue distribution
assays. Substitution with heavier isotopes, such as substitution of
one or more hydrogen atoms with deuterium (.sup.2H), can provide
pharmacological advantages in some instances, including increased
metabolic stability. Isotopically labeled compounds of formula (I)
can generally be prepared by substituting an isotopically labeled
reagent for a non-isotopically labeled reagent.
3. Formulation and Use of Compounds of the Invention
[0111] Compounds of the invention can be used for both medical
applications and animal husbandry or veterinary applications.
Typically, the compound, or a pharmaceutical composition including
the compound, is used in humans, but may also be used in other
mammals. The term "patient" is intended to denote a mammalian
individual and is so used throughout the specification and in the
claims. The primary applications of this invention involve human
patients, but this invention may be applied to laboratory, farm,
zoo, wildlife, pet, sport or other animals.
[0112] The compounds disclosed herein, or made by methods disclosed
herein, may be used for the treatment of any condition, syndrome or
disease, and in particular for any condition, syndrome or disease
for which a melanocortin receptor-specific molecule has some
efficacy. The compounds disclosed herein, or made by methods
disclosed herein, can have one or more advantages relative to
melanocortin receptor-specific peptides, including but not limited
to advantages such as increased resistance to enzymatic
degradation, increased circulation half life, increased
bioavailability, increased efficacy, increased specificity,
prolonged duration of effect and combinations of the foregoing.
[0113] Salt Form of Compounds. The compounds of this invention may
be in the form of any pharmaceutically acceptable salt. The
pharmaceutically acceptable salts may be salts prepared from
pharmaceutically acceptable non-toxic bases or acids including
inorganic or organic bases and inorganic or organic acids. Salts
derived from inorganic bases include salts of aluminum, ammonium,
calcium, copper, ferric, ferrous, lithium, magnesium, manganic,
manganous, potassium, sodium, zinc, and the like. Particularly
preferred are the ammonium, calcium, lithium, magnesium, potassium,
and sodium salts. Salts derived from pharmaceutically acceptable
organic non-toxic bases include salts of primary, secondary, and
tertiary amines, substituted amines including naturally occurring
substituted amines, cyclic amines, and basic ion exchange resins,
such as arginine, betaine, caffeine, choline,
N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol,
2-dimethylaminoethanol, ethanolamine, ethylenediamine,
N-ethyl-morpholine, N-ethylpiperidine, glucamine, glucosamine,
histidine, hydrabamine, isopropylamine, lysine, methylglucamine,
morpholine, piperazine, piperidine, polyamine resins, procaine,
purines, theobromine, triethylamine, trimethylamine,
tripropylamine, tromethamine, and the like.
[0114] When the compound of this invention is basic, acid addition
salts may be prepared from pharmaceutically acceptable non-toxic
acids, including inorganic and organic acids. Such acids include
acetic, benzenesulfonic, benzoic, camphorsulfonic, carboxylic,
citric, ethanesulfonic, formic, fumaric, gluconic, glutamic,
hydrobromic, hydrochloric, isethionic, lactic, maleic, malic,
mandelic, methanesulfonic, malonic, mucic, nitric, pamoic,
pantothenic, phosphoric, propionic, succinic, sulfuric, tartaric,
p-toluenesulfonic acid, trifluoroacetic acid, and the like. Acid
addition salts of the compound of this invention are prepared in a
suitable solvent from the compound and an excess of an acid, such
as hydrochloric, hydrobromic, sulfuric, phosphoric, acetic,
trifluoroacetic, citric, tartaric, maleic, succinic or
methanesulfonic acid. The acetate salt form is especially useful.
Where the compound of embodiments of this invention include an
acidic moiety, suitable pharmaceutically acceptable salts may
include alkali metal salts, such as sodium or potassium salts, or
alkaline earth metal salts, such as calcium or magnesium salts.
[0115] Pharmaceutical Formulations and Compositions. An embodiment
of this invention provides an oral or enteral pharmaceutical
formulation that includes a compound of this invention and one or
more stabilizing agents, preservatives, solubilizing agents,
bulking agents, wetting and solubility enhancing agents, buffering
agents, and other excipients.
[0116] In one aspect either immediate release or modified release
formulations, comprising a compound of this invention, are
employed. The modified release formulations may be an extended
release formulation or a delayed released formulation or may be a
modified release formulation that provides a pulsatile release of a
compound of this invention.
[0117] Because of their ease of administration, tablets and
capsules represent an advantageous oral dosage unit form. If
desired, a composition including a compound of this invention may
be coated by standard aqueous or nonaqueous techniques. The amount
of active compound in such therapeutically useful compositions is
such that an effective dosage will be obtained.
[0118] One embodiment of the invention provides a dosage form
comprising a pharmaceutical formulation including a compound of
this invention, where the dosage form is selected from tablets,
mini tablets, capsules, beads, granules, powders, caplets, troches,
sachets, cachets, pouches, gums, sprinkles, solutions, suspensions,
and buccal preparations. The tablets may be osmotic tablets, matrix
tablets, bi- and multilayer tablets, fast disintegrating tablets
and other type of tablets commonly used in the art. The formulation
may be also presented in the form of pellets in a capsule, where
the capsule may be swallowed whole or can be opened and the pellets
sprinkled on to soft food or in a liquid and then swallowed. The
dosage form may further comprise components such as starches,
sugars, microcrystalline cellulose, diluents, granulating agents,
lubricants, binders, disintegrating agents and the like employed in
oral solid preparations.
[0119] Any of a variety of enteric and drug release formulations
and coatings may further be employed with the selected dosage form,
such as immediate release, delayed release or sustained release
formulations and coatings. For example, poly(meth)acrylate
chemistry, such as Eudragit.RTM. polymers, may be employed to yield
the desire release profile. For coatings, such as enteric coatings,
various Eudragit.RTM. polymers may be employed to provide
gastrointestinal resistance, pH-controlled drug release,
gastrointestinal targeting, and the like.
[0120] Bulking agents may make up the largest quantity of the
dosage form, and include agents such as lactose anhydrous or
lactose monohydrate, glyceryl behenate, hypromellose, ascorbic
acid, benzoic acid, gelatin, carbomer, low moisture
microcrystalline cellulose, colloidal silicon dioxide, dextrates
(anhydrous), dextrose (anhydrous), maltol, fructose, glyceryl
palmitostearate, glyceryl monostearate, guar gum, lactilol
(anhydrous), magnesium carbonate, maltitol, maltose, mannitol,
polyethylene glucol, polyethylene oxide, sodium citrate sorbitol,
sucrose, compressible sugar, confectioner's sugar, xylitol and the
like.
[0121] The selected dosage form, such as tablets, pills, capsules,
and the like, may contain a binder such as gum tragacanth, acacia,
corn starch or gelatin; excipients such as dicalcium phosphate; a
disintegrating agent such as corn starch, potato starch or alginic
acid; a lubricant such as magnesium stearate; and a sweetening
agent such as sucrose, lactose or saccharin. When a dosage unit
form is a capsule, it may contain, in addition to materials of the
above type, a liquid carrier such as a fatty oil.
[0122] Various other materials may be utilized as coatings or to
modify the physical form of the dosage unit. For instance, tablets
may be coated with shellac, sugar or both. A syrup or elixir may
contain, in addition to the active ingredient, sucrose as a
sweetening agent, methyl and propylparabens as preservatives, a dye
and a flavoring such as cherry or orange flavor.
[0123] Compounds of this invention may also be administered
parenterally. Solutions or suspensions of active compounds can be
prepared in water suitably mixed with a surfactant such as
hydroxy-propylcellulose. Dispersions can also be prepared in
glycerol, liquid polyethylene glycols and mixtures thereof in oils.
These preparations may optionally contain a preservative to prevent
the growth of microorganisms. Lyophilized single unit formulations
may also be employed, such as are reconstituted with saline prior
to administration, and thus do not require a preservative.
[0124] The pharmaceutical forms suitable for injectable use include
sterile aqueous solutions or dispersions and sterile powders, such
as lyophilized formulations, for the extemporaneous preparation of
sterile injectable solutions or dispersions. In all cases, the form
must be sterile and must be fluid to the extent that it may be
administered by syringe. The form must be stable under the
conditions of manufacture and storage and must be preserved against
the contaminating action of microorganisms such as bacteria and
fungi. The carrier can be a solvent or dispersion medium
containing, for example, water, ethanol, a polyol, for example
glycerol, propylene glycol or liquid polyethylene glycol, suitable
mixtures thereof, and vegetable oils.
[0125] The compounds of this invention may in an alternative aspect
be therapeutically administered by means of an injection, typically
a deep intramuscular injection, such as in the gluteal or deltoid
muscle, of a time release injectable formulation. In one
embodiment, a compound of this invention is formulated with a PEG,
such as poly(ethylene glycol) 3350, and optionally one or more
additional excipients and preservatives, including but not limited
to excipients such as salts, polysorbate 80, sodium hydroxide or
hydrochloric acid to adjust pH, and the like. In another embodiment
a compound of this invention is formulated with a poly(ortho
ester), which may be an auto-catalyzed poly(ortho ester) with any
of a variable percentage of lactic acid in the polymeric backbone,
and optionally one or more additional excipients.
[0126] In one embodiment poly (D,L-lactide-co-glycolide) polymer
(PLGA polymer) is employed, preferably a PLGA polymer with a
hydrophilic end group, such as PLGA RG502H from Boehringer
Ingelheim, Inc. (Ingelheim, Germany). Such formulations may be
made, for example, by combining a compound of this invention in a
suitable solvent, such as methanol, with a solution of PLGA in
methylene chloride, and adding thereto a continuous phase solution
of polyvinyl alcohol under suitable mixing conditions in a reactor.
In general, any of a number of injectable and biodegradable
polymers, which are preferably also adhesive polymers, may be
employed in a time release injectable formulation. The teachings of
U.S. Pat. Nos. 4,938,763, 6,432,438, and 6,673,767, and the
biodegradable polymers and methods of formulation disclosed
therein, are incorporated here by reference. The formulation may be
such that an injection is required on a weekly, monthly or other
periodic basis, depending on the concentration and amount of
compound, the biodegradation rate of the polymer, and other factors
known to those of skill in the art. For injection or other liquid
administration formulations, water containing at least one or more
buffering constituents is preferred, and stabilizing agents,
preservatives and solubilizing agents may also be employed. For
solid administration formulations, any of a variety of thickening,
filler, bulking and carrier additives may be employed, such as
starches, sugars, amino acids, fatty acids and the like. For
topical administration formulations, any of a variety of creams,
ointments, gels, lotions and the like may be employed. For most
pharmaceutical formulations, non-active ingredients will constitute
the greater part, by weight or volume, of the preparation.
[0127] Therapeutically Effective Amount. In general, the actual
quantity of compound of this invention administered to a patient
will vary between fairly wide ranges depending upon the mode of
administration, the formulation used, and the response desired. The
dosage for treatment is administration, by any of the foregoing
means or any other means known in the art, of an amount sufficient
to bring about the desired therapeutic effect. Thus a
therapeutically effective amount includes an amount of a compound
or pharmaceutical composition of this invention that is sufficient
to induce a desired effect.
[0128] In general, the compounds of this invention are highly
active. For example, for systemic applications the compound can be
administered at about 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 50, or 100
mg/kg body weight, depending on the specific compounds selected,
the desired therapeutic response, the route of administration, the
formulation and other factors known to those of skill in the art.
For oral administration, a tablet or capsule containing from about
1 to about 1000 mg of compound is administered once daily, twice
daily, three times daily or four times daily. In one aspect, from
about 10 to about 1000 mg of the compound of Example 7.1 is
administered once or twice daily as a tablet or capsule.
[0129] Therapeutic Application. Compounds of this invention that
are ligands of MC4r are believed to be useful in treating diseases,
disorders and/or conditions responsive to modulation of the MC4r,
more particularly activation of the MC4r, i.e. diseases, disorders
and/or conditions which would benefit from agonism (including full
or partial agonism) at the MC4r, including energy homeostasis and
metabolism related (such as diabetes, in particular type 2
diabetes; dyslipidemia; fatty liver; gout; hypercholesterolemia;
hypertriglyceridemia; hyperuricacidemia; impaired glucose
tolerance; impaired fasting glucose; insulin resistance syndrome;
and metabolic syndrome), food intake related (such as hyperphagia;
binge eating; bulimia; and compulsive eating) and/or energy balance
and body weight related diseases, disorders and/or conditions, more
particularly such diseases, disorders and conditions characterized
by excess body weight and/or excess food intake. In one aspect,
compounds of the invention are utilized to treat conditions
relating to various expression or receptor genetic diseases such as
pro-opiomelanocortin deficiency due to mutations in the POMC gene
(POMC heterozygous deficiency obesity), Prader-Willi syndrome,
obesity due to MC4r deficiency, leptin receptor deficiency obesity,
leptin deficiency obesity, including congenital leptin deficiency,
Bardet Biedl syndrome, Alstrom syndrome, and various other
diseases, conditions, genetic deficiencies, metabolic disorders,
and syndromes.
[0130] It will be understood that there are medically accepted
definitions of obesity and overweight. A patient may be identified
by, for example, measuring body mass index (BMI), which is
calculated by dividing weight in kilograms by height in metres
squared, and comparing the result with the definitions. The
recommended classifications for BMI in humans, adopted by the
Expert Panel on the Identification, Evaluation and Treatment of
Overweight and Obesity in Adults, and endorsed by leading
organizations of health professionals, are as follows: underweight
<18.5 kg/m.sup.2, normal weight 18.5-24.9 kg/m.sup.2, overweight
25-29.9 kg/m.sup.2, obesity (class 1) 30-34.9 kg/m.sup.2, obesity
(class 2) 35-39.9 kg/m.sup.2, extreme obesity (class 3).gtoreq.40
kg/m.sup.2 (Practical Guide to the Identification, Evaluation, and
Treatment of Overweight and Obesity in Adults, The North American
Association for the Study of Obesity (NAASO) and the National
Heart, Lung and Blood Institute (NHLBI) 2000). Modifications of
this classification may be used for specific ethnic groups. Another
alternative for assessing overweight and obesity is by measuring
waist circumference. There are several proposed classifications and
differences in the cut-offs based on ethnic group. For instance,
according to the classification from the International Diabetes
Federation, men having waist circumferences above 94 cm (cut off
for europids) and women having waist circumferences above 80 cm
(cut off for europids) are at higher risk of diabetes,
dyslipidemia, hypertension and cardivascular diseases because of
excess abdominal fat. Another classification is based on the
recommendation from the Adult Treatment Panel III where the
recommended cut-offs are 102 cm for men and 88 cm for women.
However, the compounds of this invention may also be used for
reduction of self-diagnosed overweight and for decreasing the risk
of becoming obese due to life style, genetic considerations,
heredity and/or other factors. The term "diabetes" includes type 1
diabetes (insulin-dependent diabetes mellitus), latent autoimmune
diabetes mellitus of adults (LADA), and type 2 diabetes.
[0131] It is believed that compounds of this invention which are
MC4r agonists or partial agonists, upon administration to an
animal, including man, will reduce food intake, body weight and/or
body weight gain in that animal. Without being bound by any theory,
it is believed that such compounds of this invention act by
modulating appetite and/or satiety, increasing metabolic rate,
reducing intake of and/or craving for fat and/or carbohydrates.
[0132] Without being bound by any theory, it is also believed that
compounds of this invention which are MC4r agonists or partial
agonists, act by enhancing glucose tolerance and/or decreasing
insulin resistance. It is therefore believed that such compounds of
this invention can be useful also for treatment of type 2 diabetes
in underweight and normal weight individuals as well as in
overweight and obese individuals.
[0133] Thus compounds of this invention which are MC4r agonists or
partial agonists have utility in attenuating food intake and body
weight gain, and for treatment of obesity, diabetes mellitus type
2, metabolic syndrome and related conditions and indications, as
well as various expression or receptor genetic diseases such as
pro-opiomelanocortin deficiency due to mutations in the POMC gene
(POMC heterozygous deficiency obesity), Prader-Willi syndrome,
obesity due to MC4r deficiency, leptin receptor deficiency obesity,
leptin deficiency obesity, Bardet Biedl syndrome, Alstrom syndrome,
and various other diseases, conditions, genetic deficiencies,
metabolic disorders, and syndromes.
[0134] MC4r is a part of the leptin-melanocortin pathway, or
pro-opiomelanocortin (POMC)-MC4r pathway. Constituent members of
this pathway include a wide diversity of proteins, including
.alpha.-MSH, POMC, leptin and leptin receptors. Certain diseases,
conditions and syndromes result from mutations and variations,
including genetic defect disorders, associated with or in one or
more constituent members of the POMC-MC4r pathway. The compounds of
this invention may, as hereafter described, be useful in treatment
of diseases, conditions and syndromes resulting from mutations and
variations, including genetic defect disorders, associated with or
in one or more constituent members of the POMC-MC4r pathway.
[0135] The hypothalamic POMC-MC4r pathway is part of the regulatory
system modulating feed behavior, appetite and body weight. There
are a number of diseases, conditions and syndromes which have been
described associated with disruption of the hypothalamic POMC-MC4r
pathway which is believed to result from genetic defects or
disruptions, include defects or disruptions in genes in the
POMC-MC4r pathway. For example, Prader-Willi syndrome manifests in
significant hyperphagia and severe obesity, and may include other
features and signs, such as learning disabilities, abnormal
neurologic function, hypogonadism, short stature and developmental
and cognitive delays. The compounds of this invention may, as
hereafter described, be useful in treatment of Prader-Willi
syndrome, as well as other diseases, conditions and syndromes
involving defects or disruptions in genes in the POMC-MC4r
pathway.
[0136] Compounds of the invention may be utilized and are indicated
for the treatment of the obesity and hyperphagia associated with
POMC deficiency caused by homozygous or compound heterozygous loss
of function mutations in the POMC gene located at chromosome 2,
position 23.3. Mutations in the POMC gene that result in complete
loss of or that significantly reduce production of the POMC
polypeptide lead to no or reduced production of .alpha.-MSH. This
loss of endogenous .alpha.-MSH results in significantly diminished
MC4r activity with resulting hyperphagia and obesity. Compounds of
this invention may be utilized as a replacement MC4r agonist
therapeutic in patients with little or no endogenous
.alpha.-MSH.
[0137] For a variety of diseases, conditions or syndromes
associated with disruption of the hypothalamic POMC-MC4r pathway,
various genetic and genotyping tests may be employed as part of
diagnosis of prospective patients, and determining suitability for
use of the compounds of this invention in such prospective
patients. By way of example and not limitation, for Prader-Willi
syndrome it is possible to utilize genetic testing such as
DNA-based methylation testing to ascertain the the loss of active
genes in a specific part of chromosome 15, the 15q11-q13 region,
specifically deletion of at least the 15q11-q13 region of paternal
chromosome 15. Similarly, POMC deficiency may be diagnosed by loss
of function mutations in the POMC gene. Thus treatment with a
compound of this invention may include various diagnostic and
genetic tests to ascertain the presence of a loss of function
mutuation or other mutuation in the POMC-MC4r pathway, including
but not limited to a loss of function mutuation for Prader-Willi
syndrome affecting the 15q11-q13 region, loss of function mutations
in the POMC gene, the leptin gene, the leptin receptor gene, and
various other genes in the POMC-MC4r pathway.
4. Synthetic Methods for Compounds of the Invention
[0138] The following synthetic methods were employed in making
compounds of this invention. It is to be understood that the
invention and the compounds disclosed herein are not limited to
those made by the following methods of synthesis, but that other or
alternative methods of synthesis can be employed to make compounds
within the scope of this invention.
[0139] The following abbreviations are employed in the synthetic
schemes, and have the meanings given:
[0140] alloc--allyloxycarbonyl
[0141] Boc--tert-butoxycarbonyl
[0142] cbz--carboxybenzyl
[0143] EDC--1-ethyl-3-(3-dimethyllaminopropyl)carbodiimide
hydrochloride
[0144] Fmoc--9H-fluoren-9-ylmethoxycarbonyl
[0145] HATU--2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl
uronium hexafluorophosphate
[0146]
HBTU--O-benzotriazole-N,N,N',N'-tetramethyl-uronium-hexafluoro-phos-
phate
[0147] PG--Refers to a protecting group, including Fmoc, Boc, cbz
or alloc
[0148] TBTU--O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
tetrafluoroborate
[0149] TFA--trifluoroacetic acid
[0150] The following exemplifies the synthesis of the compound of
Example 7.1 as hereafter described, yielding HCl salt of Example
7.1 with an overall yield of 47.9%. The compounds of Examples 7.2
through 7.11, and other compounds within the scope of the
invention, may be made through similar synthetic routes, modified
as appropriate to yield the desired compound.
##STR00005## ##STR00006##
[0151] Synthesis of Compound 3: Commercially available
1-t-butyloxycarbonyl-4-amino-piperazine 1 (1.0 g, 4.97 mmol) and
N-acetyl-4-piperidone 2 (0.611 mL, 4.97 mmol) were stirred in 30 mL
of dichloromethane at room temperature for 30 minutes. To the
solution was added 1.68 g of solid sodium triacetoxyborohydride
(7.95 mmol) in portions, and the suspension was stirred at room
temperature over night. Additional sodium triacetoxyborohydride
(500 mg, 2.36 mmol) was added and stirred for 5 hours. The reaction
mixture containing product 3 was used directly in the next
step.
##STR00007##
[0152] Synthesis of Compound 4: To the crude mixture containing
compound 3 was added isobutylaldehyde (0.453 mL, 4.97 mmol) and
sodium triacetoxyborohydride (1.68 g, 7.95 mmol). The cloudy
solution was stirred at room temperature overnight. LC-MS showed an
incompleted reaction. Additional isobutylaldehyde (0.2 mL, 2.19
mmol) and sodium triacetoxyborohydride (500 mg, 2.36 mmol) were
added, and stirred for 10 hours. LC-MS showed that the reaction was
still not completed. More isobutylaldehyde (0.4 mL, 4.38 mmol) and
sodium triacetoxyborohydride (500 mg, 2.36 mmol) were added and
stirred over night again, to drive the reaction to completion. The
mixture was diluted with 50 mL of dichloromethane. The organic
layer was washed with saturated sodium bicarbonate solution
2.times., dried over sodium sulfate, filtered and concentrated to
crude product. The crude was purified by flash column with a
gradient of 50% to 100% ethyl acetate in heptane. Pure fractions
were pooled and concentrated to 1.54 g of compound 4 as yellow foam
(overall yield 81.1%).
##STR00008##
[0153] Synthesis of Compound 5: Compound 4 (1.54 g, 4.03 mmol) was
dissolved in 10 mL of dichloromethane, followed by 20 mL of 4 N HCl
solution and 10 mL of methanol. The clear solution was stirred for
2 hours, then evaporated with methanol and dichloromethane twice,
and dried under high vacuum before the next step.
[0154] The dried intermediate was dissolved in 80 mL of
dichloromethane. To this solution was added Boc-4-Cl-D-Phe (1.57 g,
5.24 mmol), N,N-Diisopropylethylamine (DIEA) (1.97 mL, 12.1 mmol),
1-Hydroxy-7-azabenzotriazole (HOAt) (5.49 mmol) and
1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium
3-oxid hexafluorophosphate (HATU) (2.14 g, 5.6 mmol). The
suspension was stirred at r.t. over night. After dilution with 50
mL of dichloromethane, the organic layer was washed with saturated
sodium bicarbonate and concentrated. Crude product was purified on
a 40 gram silica gel Isco column utilizing a gradient of 50% to
100% ethyl acetate in heptanes. Purified fractions were pooled and
concentrated.
[0155] The purified intermediate 5 was dissolved in 5 mL of
dichloromethane, followed by 10 mL of 4 N HCl solution and 5 mL of
methanol. The clear solution was stirred for 45 minutes, then
evaporated with methanol twice, followed by evaporation under high
vacuum.
##STR00009##
[0156] Synthesis of Compound 6: Compound 5 (312 mg, 0.583 mmol) and
Boc-Aib (154 mg, 0.758 mmol) were dissolved in 8 mL of
dichloromethane, followed by DIEA (0.405 mL, 2.33 mmol) to afford a
clear solution. Then HOAt (79 mg, 0.583 mmol) and HATU (332 mg,
0.874 mmol) were added, and stirred at room temperature over night.
After dilution with 40 mL of dichloromethane, the organic layer was
washed with saturated sodium bicarbonate and concentrated. The
crude product was purified by HPLC, and the combined fractions
concentrated.
[0157] The concentrated intermediate was dissolved in the mixture
of 2 mL of TFA and dichloromethane (1:1, v/v), and stirred for 30
minutes. The solvents were removed in vacuo, and the residue was
partitioned between dichloromethane and saturated ammonium
hydroxide solution. The desired product was extracted from the
basic aqueous layer with dichloromethane 4 times. 4N HCl was added
to the combined organic layer until thick precipitate formed and
the pH was approximately 1. The solvents were removed in vacuo and
the residue was dissolved in a mixture of 9 mL of t-butanol and 1
mL of water. The resulting solution was lyophilized to yield 201.53
mg of Compound 6 (the compound of Example 7.1) (yield 59.1%).
[0158] Alternate Synthesis with Diverse R.sub.1 and R.sub.5
Groups.
[0159] Any of a variety of R.sub.5 groups may be introduced to 3 by
utilizing equamolar amounts of R.sub.5-aldehyde and 3 in a solvent
such as methylene chloride, dichloroethane or tetrahydrofuran and
adding NaBH(OAc).sub.3 portionwise (typically at 1.5 times the
molar concentration of the R.sub.5-aldehyde). R.sub.5 may be a
C.sub.1 to C.sub.17 linear or branched alkyl, cycloalkyl, or
alkylcycloalkyl, or a C.sub.1 to C.sub.7 acyl group, in each
instance optionally substituted with one or more substituents, and
when one or more substituents are present, such substituents are
the same or different and independently halo, amino,
monosubstituted amino, disubstituted amino, hydroxy, or carboxy.
The resulting solution is stirred at room temperature for 16 hours,
washed with saturated sodium bicarbonate, dried over sodium sulfate
and concentrated to yield product.
[0160] The R.sub.5 group may also be introduced to 3 (1.0 mmol) by
the coupling of an organic acid (1.1 mmol), a dehydrating reagent
such as TBTU, EDC, HATU or HBTU (1.1 mmol) and an organic base such
as N-methylmorpholine or diisopropyl ethyl amine (2.0 mmol) in a
solvent such as methylene chloride, tetrahydrofuran or dioxane.
##STR00010##
[0161] Alternatively, an R.sub.5 group may be introduced to 3 by
stirring with an activated group (1.2 equivalents), such as an
organic acid chloride, isocyanate, chloroformate or sulfonyl
chloride (generically R.sub.1--X) and excess base such as pyridine,
diisopropyl ethyl amine, triethylamine or 2,4 lutidine in a solvent
such as methylene chloride, dichloroethane or tetrahydrofuran. Any
R.sub.5 may be employed; typical R.sub.5 groups include methyl,
isopropyl, isobutyl, acetyl, sulfonyl, carbamoyl and the like.
[0162] By way of example, use of Boc as a PG for piperazine and
alloc as a PG for piperidine results, following introduction of the
R.sub.5 group, in compound 10. Compound 10 is selectively
deprotected, such as by removal of the alloc group to yield
compound 11. For example, to remove the alloc group compound 11 is
treated at room temperature with catalytic
tetrakis(triphenylphosphine)palladium in a solvent such as
methylene chloride, tetrahydrofuran or dichloroethane in the
presence of an excess (10.times.) of an allyl acceptor scavenger,
such as phenylsilane or 1,3-dimethylbarbituric acid. Following
addition of the --C(.dbd.O)--R.sub.1 group and removal of the
protecting group PG (such as for example alloc, Boc or cbz), the
solution is dried over sodium sulfate and evaporated to give the
R.sub.1 and R.sub.5 substituted 12.
##STR00011##
[0163] Alternate Synthesis with Diverse R.sub.6a, R.sub.6b,
R.sub.2, R.sub.3 and R.sub.4 Groups. It is possible and
contemplated to either stepwise couple a protected, such as a Boc
protection, optionally substituted phenylalanine, where R.sub.6a
and R.sub.6b are permitted substituents, and an .alpha.,
.alpha.-disubstituted amino acid comprising R.sub.2, R.sub.3 and
R.sub.4 substituents, to R.sub.1 and R.sub.5 substituted 12, or to
alternatively to couple optionally substituted phenylalanine, where
R.sub.6a and R.sub.6b are permitted substituents, to an .alpha.,
.alpha.-disubstituted amino acid comprising R.sub.2, R.sub.3 and
R.sub.4 substituents, and thereafter to couple the resulting
compound to R.sub.1 and R.sub.5 substituted 12.
[0164] For stepwise coupling, R.sub.1 and R.sub.5 substituted 12
(1.0 mmol) is added slower to a stirred mixture of a Boc protected
optionally substituted phenylalanine, where R.sub.6a and R.sub.6b
are permitted substituents (1.1 mmol), a a dehydrating reagent such
as TBTU, EDC, HATU or HBTU (1.1 mmol) and an organic base such as
N-methylmorpholine or diisopropyl ethyl amine (2.0 mmol) in a
solvent such as methylene chloride, tetrahydrofuran or dioxane.
After stirring for 16 hours the solution is extracted into a
solvent such as methylene chloride or ethyl acetate and washed with
saturated sodium bicarbonate, dried over sodium sulfate and
concentrated. De-protection is accomplished by stirring with excess
TFA in solvent such as methylene chloride for one hour followed by
evaporation of the solvent. The resulting oil is basified to pH=8
with sodium bicarbonate and extracted into a solvent such as
methylene chloride or ethyl acetate. The solution is dried over
sodium sulfate and evaporated to yield an oil as compound 13.
##STR00012##
[0165] Compound 13 (1.0 mmol) is added slowly to a stirred mixture
of a protected, such as a Boc-protected, .alpha.-disubstituted
amino acid comprising R.sub.2, R.sub.3 and R.sub.4 substituents
(1.1 mmol), a dehydrating reagent such as TBTU, EDC, HATU or HBTU
(1.1 mmol) and an organic base such as N-methylmorpholine or
diisopropyl ethyl amine (2.0 mmol) in a solvent such as methylene
chloride, tetrahydrofuran or dioxane. After stirring for 16 hours
the solution is extracted in a solvent such as methylene chloride
or ethyl acetate and washed with saturated sodium bicarbonate,
dried over sodium sulfate and concentrated. Deprotection is
accomplished by stirring with excess TFA in solvent such as
methylene chloride for one hour followed by evaporation of the
solvent. The residue is dissolved in an appropriate solvent, such
as 35% aqueous methanol, and purified by HPLC to yield compound 14.
The resulting compound 14 is purified by HPLC.
##STR00013##
[0166] R.sub.6a and R.sub.6b may be optionally substituted with one
or more substituents, including specifically substitutents that are
the same or different and independently alkyl, haloalkyl,
cycloalkyl, alkoxy, alkythio, halo, nitro, acyl, cyano, aryl,
alkylaryl, aryloxy, amino, monosubstituted amino, disubstituted
amino, sulfonamide, hydroxy, carboxy, or alkoxy-carbonyl. As
appropriate, reactive substituents may include protecting groups,
which are deprotected to yield final product. Protecting groups
other than Boc, including specifically alloc, may be employed.
[0167] In the foregoing methods and synthetic schemes, any
optionally protected and optionally substituted phenylalanine,
where R.sub.6a and R.sub.6b are permitted substituents, may be
employed. D-isomer phenylalanine (D-Phe) is preferred, but L-isomer
phenylalanine may be employed. Preferred is PG-D-Phe optionally
substituted with one or two substitutents, which substituents may
be the same or different. Substituents include, but are not limited
to, --Cl, --CF.sub.3, --CN, --CH.sub.3, --OH, --NO.sub.2, --F,
--Br, --Cl, --I, --OCH.sub.3 and the like. Thus side chains of Phe,
preferably where Phe is the D-isomer, may include, but are not
limited to, the following:
TABLE-US-00001 Abbreviation Common Name Side Chain Phe(2-CF.sub.3)
2-trifluoromethyl phenylalanine ##STR00014## Phe(2-C(=O)--NH.sub.2)
2-carbamoyl- phenylalanine ##STR00015## Phe(2-Me) 2-methyl
phenylalanine ##STR00016## Phe(2-CN) 2-cyano phenylalanine
##STR00017## Phe(2-Cl) 2-chloro phenylalanine ##STR00018##
Phe(2,4-diCl) 2,4-dichloro phenylalanine ##STR00019## Phe(2,4-diMe)
2,4-dimethyl phenylalanine ##STR00020## Phe(2-F) 2-flouro
phenylalanine ##STR00021## Phe(2-NO.sub.2) 2-nitro phenylalanine
##STR00022## Phe(3-CF.sub.3) 3-triflouromethyl phenylalanine
##STR00023## Phe(3-C(=O)--NH.sub.2) 3-carbamoyl- phenylalanine
##STR00024## Phe(3-CN) 3-cyano phenylalanine ##STR00025## Phe(3-Cl)
3-chloro phenylalanine ##STR00026## Phe(3,4-diCl) 3,4-dichloro
phenylalanine ##STR00027## Phe(3-F) 3-fluoro phenylalanine
##STR00028## Phe(3,4-diF) 3,4-difluoro phenylalanine ##STR00029##
Phe(3,5-diF) 3,5-difluoro phenylalanine ##STR00030## Phe(3-Me)
3-methyl phenylalanine ##STR00031## Phe(3-NO.sub.2) 3-nitro
phenylalanine ##STR00032## Phe(3,4-diOMe) 3,4-dimethoxy
phenylalanine ##STR00033## Phe(4-C(=O)--NH.sub.2) 4-carbamoyl-
phenylalanine ##STR00034## Phe(4-Me) 4-methyl phenylalanine
##STR00035## Phe(4-CF.sub.3) 4-trifluoromethyl phenylalanine
##STR00036## Phe(4-CN) 4-cyano phenylalanine ##STR00037## Phe(4-Cl)
4-chloro phenylalanine ##STR00038## Phe(4-F) 4-fluoro phenylalanine
##STR00039## Phe(4-NH.sub.2) 4-amino phenylalanine ##STR00040##
Phe(4-NO.sub.2) 4-nitro phenylalanine ##STR00041## Phe(4-Ph)
4-phenyl phenylalanine ##STR00042## Phe(4-OMe) 4-methoxy
phenylalanine ##STR00043## Phe(4-tBu) 4-tert butyl phenylalanine
##STR00044##
[0168] In the foregoing methods and synthetic schemes, any .alpha.,
.alpha.-disubstituted amino acid may conveniently be employed to
provide the R.sub.2, R.sub.3 and R.sub.4 groups. In general, an
".alpha., .alpha.-disubstituted amino acid" includes any
.alpha.-amino acid having a further substituent in the
.alpha.-position, which substituent may be the same as or different
from the side chain moiety of the .alpha.-amino acid. For purposes
of the specification and claims, either a "substituent" or the
"side chain moety" of an .alpha., .alpha.-disubstituted amino acid
is referred to as a side chain, such that an .alpha.,
.alpha.-disubstituted amino acid includes two side chains. Suitable
substituents, in addition to the side chain moiety of the
.alpha.-amino acid, include but are not limited to C.sub.1 to
C.sub.6 linear or branched alkyl. Aib is an example of an .alpha.,
.alpha.-disubstituted amino acid. While .alpha.,
.alpha.-disubstituted amino acids can be referred to using
conventional L- and D-isomeric references, it is to be understood
that such references are for convenience, and that where the
substituents at the .alpha.-position are different, such amino acid
can interchangeably be referred to as an .alpha.,
.alpha.-disubstituted amino acid derived from the L- or D-isomer,
as appropriate, of a residue with the designated amino acid side
chain moiety. Thus (S)-2-Amino-2-methyl-hexanoic acid can be
referred to as either an .alpha., .alpha.-disubstituted amino acid
derived from L-Nle or as an .alpha., .alpha.-disubstituted amino
acid derived from D-Ala. Whenever an .alpha., .alpha.-disubstituted
amino acid is provided, it is to be understood as including all (R)
and (S) configurations thereof. The .alpha., .alpha.-disubstituted
amino acid at position Aaa.sup.4 may be any .alpha.,
.alpha.-disubstituted amino acid. By way of example and not
limitation, the following are included in the definition of an
.alpha., .alpha.-disubstituted amino acid which provides R.sub.2,
R.sub.3 and R.sub.4 groups.
##STR00045## ##STR00046##
5. Assay Systems for Compounds
[0169] Selected compounds are tested in assays to determine binding
and functional status and are tested in animal models of feeding
behavior as discussed below. The following assays and animal models
are employed, with modifications, if any, as discussed in the
examples.
[0170] Competitive Inhibition Assay Using
[I.sup.125]-NDP-.alpha.-MSH. A competitive inhibition binding assay
is performed using membrane homogenates prepared from HEK-293 cells
that express recombinant hMC1ra, hMC4r, hMC3r, or hMC5r, and from
B16-F10 mouse melanoma cells (containing endogenous MC1r). In the
examples that follow, all values are for human recombinant
receptors unless otherwise noted. Assays are performed in 96 well
GF/B Millipore multiscreen filtration plates (MAFB NOB10)
pre-coated with 0.5% bovine serum albumin (Fraction V). Membrane
homogenates are incubated with 0.2 nM (for hMC4r) 0.4 nM (for MC3r
and MC5r) or 0.1 nM (for mouse B16-F10 MC1r or hMC1ra)
[I.sup.125]-NDP-.alpha.-MSH (Perkin Elmer) and increasing
concentrations of test compounds in buffer containing 25 mM HEPES
buffer (pH 7.5) with 100 mM NaCl, 2 mM CaCl.sub.2, 2 mM MgCl.sub.2,
0.3 mM 1,10-phenanthroline, and 0.2% bovine serum albumin. After
incubation for 60 to 90 minutes at 37.degree. C., the assay mixture
is filtered and the membranes washed three times with ice-cold
buffer. Filters are dried and counted in a gamma counter for bound
radioactivity.
[0171] Non-specific binding is measured by inhibition of binding of
[I.sup.125]-NDP-.alpha.-MSH in the presence of 1 .mu.M
NDP-.alpha.-MSH. Maximal specific binding (100%) is defined as the
difference in radioactivity (cpm) bound to cell membranes in the
absence and presence of 1 .mu.M NDP-.alpha.-MSH. Radioactivity
(cpm) obtained in the presence of test compounds is normalized with
respect to 100% specific binding to determine the percent
inhibition of [I.sup.125]-NDP-.alpha.-MSH binding. Each assay is
conducted in duplicate or triplicate and the actual mean values are
described, with results less than 0% reported as 0%. Ki values for
test compounds are determined using Graph-Pad Prism.RTM.
curve-fitting software.
[0172] Competitive Binding Assay Using Eu-NDP-.alpha.-MSH.
Alternatively, a competitive inhibition binding assay is performed
employing Eu-NDP-.alpha.-MSH (PerkinElmer Life Sciences catalog No.
AD0225) with determination by time-resolved fluorometry (TRF) of
the lanthanide chelate. In comparison studies with
[I.sup.125]-NDP-.alpha.-MSH, the same values, within experimental
error ranges, were obtained for percent inhibition and Ki.
Typically competition experiments to determine Ki values were
conducted by incubating membrane homogenates prepared from HEK-293
cells that express recombinant hMC4r with 9 different
concentrations of test compounds of interest and 2 nM of
Eu-NDP-.alpha.-MSH in a solution containing 25 mM HEPES buffer with
100 mM NaCl, 2 mM CaCl.sub.2, 2 mM MgCl.sub.2 and 0.3 mM
1,10-phenanthroline. After incubation for 90 minutes at 37.degree.
C., the reaction was stopped by filtration over Acro Well 96-well
filter plates (Pall Life Sciences). The filter plates were washed 4
times with 200 .mu.L of ice-cold phosphate-buffered saline. DELFIA
Enhancement solution (PerkinElmer Life Sciences) was added to each
well. The plates were incubated on a shaker for 15 minutes and read
at 340 nm excitation and 615 nm emission wavelengths. Each assay
was conducted in duplicate and mean values were utilized. Ki values
were determined by curve-fitting with Graph-Pad Prism.RTM. software
using a one-site fixed-slope competition binding model.
[0173] Assay for Agonist Activity. Accumulation of intracellular
cAMP was examined as a measure of the ability of the peptides of
the present invention to elicit a functional response in HEK-293
cells that express MC4-R. Confluent HEK-293 cells that express
recombinant hMC4-R were detached from culture plates by incubation
in enzyme-free cell dissociation buffer. Dispersed cells were
suspended in Hanks' Balanced Salt Solution containing 10 mM HEPES
(pH 7.5), 1 mM MgCl.sub.2, 1 mM glutamine, 0.5% albumin and 0.3 mM
3-isobutyl-1-methyl-xanthine (IBMX), a phosphodiesterase inhibitor.
The cells were plated in 96-well plates at a density of
0.5.times.10.sup.5 cells per well and pre-incubated for 10 minutes.
Cells were exposed for 15 minutes at 37.degree. C. to peptides of
the present invention dissolved in DMSO (final DMSO concentration
of 1%) at a concentration range of 0.05-5000 nM in a total assay
volume of 200 .mu.L. NDP-.alpha.-MSH was used as the reference
agonist. cAMP levels were determined by an HTRF.RTM. cAMP
cell-based assay system from Cisbio Bioassays utilizing
cryptate-labeled anti-cAMP and d2-labeled cAMP, with plates read on
a Perkin-Elmer Victor plate reader at 665 and 620 nM. Data analysis
was performed by nonlinear regression analysis with Graph-Pad
Prism.RTM. software. The maximum efficacies of the test peptides of
the present invention were compared to that achieved by the
reference melanocortin agonist NDP-.alpha.MSH.
[0174] Low Receptor Density Systems. In functional studies
preferably a human MC4r expression system approximating maximal
human physiologic receptor densities is employed. In one aspect, a
tetracycline-regulated mammalian expression system that uses
regulatory elements from the E. coli Tn10-encoded tetracycline
(Tet) resistance operon was employed (T-REx.TM. System,
Invitrogen). By use of the T-REx.TM. System, expression of the gene
of interest, the human MC4-R gene, was repressed in the absence of
tetracycline or doxycycline and induced in the presence of
tetracycline or doxycycline, with receptor density being dependent
on the concentration of tetracycline or doxycycline. See generally
Yao F. et al: Tetracycline repressor, tetR, rather than the
tetR-mammalian cell transcription factor fusion derivatives,
regulates inducible gene expression in mammalian cells. Hum. Gene
Ther. 9:1939-1950 (1998), incorporated here by reference.
Regulation was based on the binding of tetracycline or doxycycline,
a derivative of tetracycline, to the Tet repressor and derepression
of the promoter controlling expression of the human MC4-R gene. In
general tetracycline or related regulators, such as
tetracycline-derivatives, may be used to induce the gene
expression. An inducible expression plasmid for expression of human
MC4-R gene under the control of the strong human cytomegalovirus
immediate-early (CMV) promoter and two tetracycline operator 2
(TetO.sub.2) sites was employed, together with a regulatory
plasmid, pcDNA6/TR, which encodes the Tet repressor (TetR) under
the control of the human SV40 promoter. Thus, expression of the
human MC4-R gene from the inducible expression vector was
controlled by the strong CMV promoter into which 2 copies of the
tet operator 2 (TetO.sub.2) sequence had been inserted in tandem.
The TetO.sub.2 sequences consisted of 2 copies of 19-nucleotide
sequence separated by a 2 base pair spacer. Each 19 nucleotide
TetO.sub.2 sequence served as the binding site for 2 molecules of
the Tet repressor. The expression vector pcDNA4/TO/MC4R was
introduced into TREx-293 (Invitrogen, R710-07), a cell line stably
expressing the tetracycline repressor pcDNA6/TR using lipofectamin
2000 reagent (Invitrogen, 11668-019). Alternatively, both the
expression vector and repressor are introduced into host cells by
standard transformation or transfection methods.
[0175] The receptor density at different doxycycline concentrations
was quantified in terms of binding per mg of protein derived from
cell membranes using [I.sup.125]-NDP-.alpha.-MSH in receptor
binding saturation studies to determine a B.sub.max value. The
table below quantifies the receptor density:
TABLE-US-00002 Doxycycline B.sub.max .+-. SD K.sub.D(app) .+-. SD
(ng/mL) (fmol/mg) (pM) n 0.1 62 .+-. 13 114 .+-. 57 8 1 207 .+-.
128 118 .+-. 37 11 10 10113 .+-. 2538 139 .+-. 41 11
[0176] By comparison, transformed or transfected cells not under
control of a tetracycline- or doxycycline-regulated expression
system, such as HEK-293 cells that express recombinant hMC4r used
for [I.sup.125]-NDP-.alpha.-MSH competitive inhibition assays,
typically resulted in a B.sub.max value of approximately 400
fmol/mg using [I.sup.125]-NDP-.alpha.-MSH. A receptor density
B.sub.max value of between approximately 80 and 20 fmol/mg using
[I.sup.125]-NDP-.alpha.-MSH, preferably between approximately 80
and 40 fmol/mg using [I.sup.125]-NDP-.alpha.-MSH, is a low density
receptor system that approximates maximal human physiologic
receptor densities.
6. Combination Therapy
[0177] Combination Therapy for Diabetes and/or Weight Regulation.
One or more compounds of this invention may be combined with at
least one other pharmacologically active agent that is useful in
the treatment of diabetes, such as other anti-diabetic drugs. One
or more compounds of the invention may also be combined with at
least one other pharmacologically active agent that is useful in
the treatment of obesity and/or overweight, such as other
anti-obesity drugs that affect energy expenditure, glycolysis,
gluconeogenesis, glucogenolysis, lipolysis, lipogenesis, fat
absorption, fat storage, fat excretion, hunger and/or satiety
and/or craving mechanisms, appetite/motivation, food intake, or
gastrointestinal motility.
[0178] One or more compounds of this invention may in addition or
alternatively further be combined with at least one other
pharmacologically active agent that is useful in the treatment of
diseaeses, disorders and/or conditions associated with obesity
and/or overweight, such as insulin resistance; impaired glucose
tolerance; type 2 diabetes; metabolic syndrome; dyslipidemia
(including hyperlipidemia); hypertension; heart disorders (e.g.
coronary heart disease, myocardial infarction); cardiovascular
disorders; non-alcoholic fatty liver disease (including
non-alcoholic steatohepatitis); joint disorders (including
secondary osteoarthritis); gastroesophageal reflux; sleep apnea;
atherosclerosis; stroke; macro and micro vascular diseases;
steatosis (e.g. in the liver); gall stones; and gallbladder
disorders.
[0179] According to an additional aspect of the invention there is
provided a combination treatment comprising the administration of a
pharmacologically effective amount of a compound of this invention,
or a pharmaceutically acceptable salt thereof, optionally together
with a pharmaceutically acceptable diluent or carrier, with the
simultaneous, sequential or separate administration one or more of
the following agents selected from: [0180] insulin and insulin
analogues; [0181] insulin secretagogues, including sulphonylureas
(e.g. glipizide) and prandial glucose regulators (sometimes called
"short-acting secretagogues"), such as meglitinides (e.g.
repaglinide and nateglinide); [0182] agents that improve incretin
action, for example dipeptidyl peptidase IV (DPP-4) inhibitors
(e.g. vildagliptin, saxagliptin, and sitagliptin), and
glucagon-like peptide-1 (GLP-1) agonists (e.g. exenatide); [0183]
insulin sensitising agents including peroxisome proliferator
activated receptor gamma (PPAR.gamma.) agonists, such as
thiazolidinediones (e.g. pioglitazone and 20 rosiglitazone), and
agents with any combination of PPAR alpha, gamma and delta
activity; [0184] agents that modulate hepatic glucose balance, for
example biguanides (e.g. metformin), fructose 1,6-bisphosphatase
inhibitors, glycogen phopsphorylase inhibitors, glycogen synthase
kinase inhibitors, and glucokinase activators; [0185] agents
designed to reduce/slow the absorption of glucose from the
intestine, such as alpha-glucosidase inhibitors (e.g. miglitol and
acarbose); [0186] agents which antagonise the actions of or reduce
secretion of glucagon, such as amylin analogues (e.g. pramlintide);
[0187] agents that prevent the reabsorption of glucose by the
kidney, such as sodium dependent glucose transporter 2 (SGLT-2)
inhibitors (e.g. dapagliflozin); [0188] agents designed to treat
the complications of prolonged hyperglycaemia, such as aldose
reductase inhibitors (e.g. epalrestat and ranirestat); and agents
used to treat complications related to micro-angiopathies; [0189]
anti-dyslipidemia agents, such as HMG-CoA reductase inhibitors
(statins, e.g. rosuvastatin) and other cholesterol-lowering agents;
PPAR.alpha. agonists (fibrates, e.g. gemfibrozil and fenofibrate);
bile acid sequestrants (e.g.cholestyramine); cholesterol absorption
inhibitors (e.g. plant sterols (i.e. phytosterols), synthetic
inhibitors); cholesteryl ester transfer protein (CETP) inhibitors;
inhibitors of the ileal bile acid transport system (IBAT
inhibitors); bile acid binding resins; nicotinic acid (niacin) and
analogues thereof; anti-oxidants, such as probucol; and omega-3
fatty acids; [0190] antihypertensive agents, including adrenergic
receptor antagonists, such as beta blockers (e.g. atenolol), alpha
blockers (e.g. doxazosin), and mixed alpha/beta blockers (e.g.
labetalol); adrenergic receptor agonists, including alpha-2
agonists (e.g. clonidine); angiotensin converting enzyme (ACE)
inhibitors (e.g. lisinopril), calcium channel blockers, such as
dihydropyridines (e.g. nifedipine), phenylalkylamines (e.g.
verapamil), and benzothiazepines (e.g. diltiazem); angiotensin II
receptor antagonists (e.g. candesartan); aldosterone receptor
antagonists (e.g. eplerenone); centrally acting adrenergic drugs,
such as central alpha agonists (e.g. clonidine); and diuretic
agents (e.g. furosemide); [0191] haemostasis modulators, including
antithrombotics, such as activators of fibrinolysis; thrombin
antagonists; factor Vila inhibitors; anticoagulants, such as
vitamin K antagonists (e.g. warfarin), heparin and low molecular
weight analogues thereof, factor Xa inhibitors, and direct thrombin
inhibitors (e.g. argatroban); [0192] antiplatelet agents, such as
cyclooxygenase inhibitors (e.g. aspirin), adenosine diphosphate
(ADP) receptor inhibitors (e.g. clopidogrel), phosphodiesterase
inhibitors (e.g. cilostazol), glycoprotein IIB/IIA inhibitors (e.g.
tirofiban), and adenosine reuptake inhibitors (e.g. dipyridamole);
[0193] anti-obesity agents, such as appetite suppressant (e.g.
ephedrine), including noradrenergic agents (e.g. phentermine) and
serotonergic agents (e.g. sibutramine), pancreatic lipase
inhibitors (e.g. orlistat), microsomal transfer protein (MTP)
modulators, diacyl glycerolacyltransferase (DGAT) inhibitors, and
cannabinoid (CB1) receptor antagonists (e.g. rimonabant); [0194]
feeding behavior modifying agents, such as orexin receptor
modulators and melanin-concentrating hormone (MCH) modulators;
[0195] glucagon like peptide-1 (GLP-1) receptor modulators; [0196]
neuropetideY (NPY)/NPY receptor modulators; [0197] pyruvate
dehydrogenase kinase (PDK) modulators; [0198] serotonin receptor
modulators; [0199] leptin/leptin receptor modulators; [0200]
ghrelin/ghrelin receptor modulators; [0201] monoamine
transmission-modulating agents, such as selective serotonin
reuptake inhibitors (SSRI), noradrenaline reuptake inhibitors
(NARI), noradrenalineserotonin reuptake inhibitors (SNRI), triple
monoamine reuptake blockers (e.g. tesofensine), and monoamine
oxidase inhibitors (MAOI); or a pharmaceutically acceptable salt,
solvate, solvate of such a salt or a prodrug thereof, optionally
together with a pharmaceutically acceptable carrier to a mammal,
such as man, in need of such therapeutic treatment.
[0202] In one aspect of this invention there is provided a
combination treatment comprising the administration of a
pharmacologically pharmacologically effective amount of a compound
of this invention, or a pharmaceutically acceptable salt thereof,
optionally together with a pharmaceutically acceptable carrier,
with the simultaneous, sequential or separate administration of
very low calorie diets (VLCD) or low-calorie diets (LCD).
[0203] In an additional aspect of the invention, there is provided
a method of treating obesity and/or overweight and therewith
associated complications in a mammal, such as man, in need of such
treatment which comprises administering to said animal a
pharmacologically effective amount of a compound of this invention,
or a pharmaceutically acceptable salt thereof, optionally together
with a pharmaceutically acceptable carrier, in simultaneous,
sequential or separate administration with a pharmacologically
effective amount of a compound from one of the other classes of
compounds described in this combination section, or a
pharmaceutically acceptable salt, solvate, solvate of such a salt
or a prodrug thereof, optionally together with a pharmaceutically
acceptable carrier.
[0204] In an additional aspect of the invention, there is provided
a method of treating diabetes, in particular type 2 diabetes, in a
mammal, such as man, in need of such treatment which comprises
administering to said animal a pharmacologically effective amount
of a compound of this invention, or a pharmaceutically acceptable
salt thereof, optionally together with a pharmaceutically
acceptable carrier, in simultaneous, sequential or separate
administration with a pharmacologically effective amount of a
compound from one of the other classes of compounds described in
this combination section, or a pharmaceutically acceptable salt,
solvate, solvate of such a salt or a prodrug thereof, optionally
together with a pharmaceutically acceptable carrier.
[0205] According to a further aspect of the invention there is
provided a pharmaceutical composition which comprises a compound of
this invention, or a pharmaceutically acceptable salt thereof, and
a compound from one of the other classes of compounds described in
this combination section or a pharmaceutically acceptable salt,
solvate, solvate of such a salt or a prodrug thereof, in
association with a pharmaceutically acceptable carrier.
[0206] According to a further aspect of this invention there is
provided a kit comprising a compound of this invention, or a
pharmaceutically acceptable salt thereof, and a compound from one
of the other classes of compounds described in this combination
section or a pharmaceutically acceptable salt, solvate, solvate of
such a salt or a prodrug thereof.
[0207] Combination Therapy with GLP-1 Receptor Agonists. Glucagon
analogs and compounds related to glucagon, including
pre-proglucagon, glucagon, glucagon-like peptide-1 (GLP-1),
glucagon-like peptide-2 (GLP-2) and oxyntomodulin (OXM), are known
in the art. Pre-proglucagon is a 179 amino acid precursor
polypeptide. Proglucagon is a 158 amino acid precursor polypeptide
that is differentially processed in vivo to form glucagon, GLP-1,
GLP-2, and OXM. GLP-1, GLP-2 and OXM molecules are involved in a
wide variety of physiological functions, including glucose
homeostasis, insulin secretion, gastric emptying and intestinal
growth, as well as regulation of food intake.
[0208] Glucagon is a 29-amino acid peptide that corresponds to
amino acids 53 to 81 of proglucagon. OXM is a 37 amino acid peptide
which includes the complete 29 amino acid sequence of glucagon with
an octapeptide carboxyterminal extension (amino acids 82 to 89 of
pre-proglucagon). Human GLP-1 is a 30- or 31-amino acid peptide,
generally C-terminally amidated, of the structure
H-His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-
-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-NH.sub.2 (30 amino
acid, with the 31 amino acid variant having a C-terminal Gly-OH).
GLP-1 is believed to increase insulin secretion from the pancreas
in a glucose-dependent manner; decrease glucagon secretion from the
pancreas by engagement of a specific G protein-coupled receptor;
increase insulin-sensitivity in both alpha cells and beta cells;
increase beta cells mass and insulin gene expression,
post-translational processing and incretion; inhibit acid secretion
and gastric emptying in the stomach; decrease food intake by
increasing satiety in the brain; and promote insulin
sensitivity.
[0209] There are a number of GLP-1 receptor agonists which have
been approved for pharmaceutical use in one or more, including
exenatide, sold under the trade names Byetta.RTM. and
Bydureon.RTM.; liraglutide, sold under the trade names Victoza.RTM.
and Saxenda.RTM.; lixisenatide, sold under the trade name
Lyxumia.RTM.; albiglutide, sold under the trade name Tanzeum.RTM.;
and dulaglutide, sold under the trade name Trulicity.RTM..
Exenatide is an incretin mimetic which is a synthetic version of
exendin-4, a hormone found in the saliva of the Gila monster.
Liraglutide, also an incretin mimetic, is a derivative of GLP-1.
Lixisenatide, also an incretin mimetic, is derived from the first
39 amino acids in the sequence of exendin-4, omitting proline at
position 38 and adding six lysine residues. Albiglutide is a
dipeptidyl peptidase-4-resistant GLP-1 dimer fused to human
albumin. Dulaglutide is a fusion protein consisting of two
identical, disulfide-linked chains, with each chain including a
modified N-terminal GLP-1 analog sequence covalently linked to the
Fc portion of a modified human immunoglobulin heavy chain by a
peptide linker. The foregoing approved GLP-1 receptor agonists are
all peptides or small proteins, and are administered typically by
subcutaneous injection. Some require daily injections; others can
be injected at weekly or longer intervals.
[0210] Preferred GLP-1 receptor agonists with utility in the
current invention include exenatide, liraglutide, lixisenatide,
albiglutide, and dulaglutide, all of which are approved
pharmaceutical drugs in the United States or elsewhere in the
world. Other naturally expressed peptides which bind to the GLP-1
receptor may be employed in the invention, and are to be considered
for purposes of this invention as GLP-1 receptor agonists,
including pre-proglucagon, glucagon, GLP-1, GLP-2 and OXM. Other
peptides and derivatives and modifications of peptides which may be
employed in the current invention include, but are not limited to,
those peptides and other compounds disclosed in WO2006/134340,
WO2007/100535, WO2008/10101, WO2008/152403, WO2009/155257,
WO2009/155258, WO2010/070252, WO2010/070253, WO2010/070255,
WO2010/070251, WO2011/006497, WO2011/160630, WO2011/160633,
WO2013/092703, WO2014/041195, WO2015/055802, WO2015149627,
WO2015/155139, WO2015/155140, WO2015/155141, WO2015/155151,
WO2015/193378, WO2015/193381, WO2016/0154014, and
WO2016/046753.
[0211] Thus in one aspect, the invention provides a method of
treating a patient with obesity, diabetes or metabolic syndrome,
comprising administering to the patient (a) a pharmaceutical
composition comprising a compound of formula I in a quantity
sufficient to induce at least minimal weight loss when administered
as a monotherapy not in conjunction with a GLP-1 receptor agonist
and (b) a GLP-1 receptor agonist in a quantity sufficient to induce
glycemic control but not weight loss when administered as a
monotherapy not in conjunction with the compound of formula I.
Preferably the method elicits a synergistic effect on treatment of
obesity. The quantity and schedule of administration of the
compound of formula I and the GLP-1 receptor agonist may together
be sufficient to produce synergistic effect in the treatment of
obesity. Alternatively, or additionally, the method elicits a
synergistic effect on treatment of glycemic control. The quantity
and schedule of administration of the compound of formula I and the
GLP-1 receptor agonist may together be sufficient to produce
synergistic effect in the treatment of glycemic control. In the
method, the compound of formula I may be the compound of formula
II, optionally administered by oral means, and the GLP-1 receptor
agonist may be administered by subcutaneous injection. The GLP-1
receptor agonist may be liraglutide or exenatide administered daily
or twice daily, or may be lixisenatide, albiglutide, dulaglutide or
an extended release formulation of exenatide or liraglutide
administered at weekly or greater intervals. In the method, either
or both the compound of formula I and the GLP-1 may be
sustained-release, including a sustained-released GLP-1 receptor
agonist with a duration of action of at least about twenty-four
hours, at least about one week or at least about two weeks.
[0212] In another aspect, the invention provides a method of
decreasing side effects associated with therapeutic agents for
treatment of obesity, diabetes or metabolic syndrome in a patient,
comprising:
[0213] administration of a quantity of a pharmaceutical composition
comprising a compound of formula I, wherein the quantity of the
compound of formula I administered, if administered as a
monotherapy not in conjunction with GLP-1 receptor agonist, is not
sufficient to initiate the desired pharmacological response in
treating at least one condition from the group comprising obesity,
diabetes and metabolic syndrome in the patient when administered as
a monotherapy; and [0214] administration of a quantity of GLP-1
receptor agonist, wherein the quantity of GLP-1 receptor agonist
administered, if administered as a monotherapy not in conjunction
with the compound of formula I, is not sufficient to initiate the
desired pharmacological response in treating at least one condition
from the group comprising obesity, diabetes and metabolic syndrome
in the patient when administered as a monotherapy; [0215] wherein
the quantity of the compound of formula I and the quantity of GLP-1
receptor agonist are together effective to initiate the desired
pharmacological response treating at least one condition from the
group comprising obesity, diabetes and metabolic syndrome in the
patient, [0216] thereby reducing side effects in the treatment of
at least one of obesity, diabetes or metabolic syndrome in the
patient.
[0217] In the practice of this method, the quantity of MC4r agonist
administered is in one aspect not sufficient to initiate the
desired pharmacological response of inducing weight loss. In
another aspect, the quantity of MC4r agonist administered is
sufficient to induce minimal weight loss.
[0218] The desired pharmacological response may be inducing weight
loss or inducing glycemic control.
[0219] In another aspect, the invention provides a method of
treating obesity or inducing weight loss in an obese patient,
comprising the steps of: [0220] establishing a dose of a GLP-1
receptor agonist which induces glycemic control in the patient but
which induces no more than minimal weight loss in the patient;
[0221] administration of the dose of the GLP-1 receptor agonist;
and [0222] administration of a a pharmaceutical composition
comprising a compound of formula I; [0223] wherein the dose of the
GLP-1 receptor agonist and the quantity of compound of formula I
are effective to treat obesity or to induce weight loss.
[0224] In this method, the dose of GLP-1 receptor agonist
preferably does not induce minimal weight loss in the patient. The
GLP-1 receptor agonist may be administered by subcutaneous
injection, such as liraglutide or exenatide administered daily or
twice daily or lixisenatide, albiglutide, dulaglutide or an
extended release formulation of exenatide or liraglutide
administered at weekly or greater intervals. In the method,
administration of the dose of the GLP-1 receptor agonist and
administration of a MC4r agonist may elicit a synergistic effect on
treatment of obesity, or may elicit a synergistic effect on
inducing glycemic control, or both.
[0225] Combination Therapy with Phosphodiesterase Inhibitors. For
certain applications and indications, it is desirable to increase
production of and maintain levels of cyclic adenoise 3',5'
monophosphate (cAMP), a nucleotide messenger associated with
inflammatory cell activity. Certain compounds of this invention
increase intracellular levels of cAMP, and can be coadministered
with compounds or substances that inhibit the degradation of cAMP.
cAMP is hydrolyzed to an inactive form by phosphodiesterase (PDE);
compounds or substances that inhibit PDE may thereby result in
maintenance of and/or an increase in available cAMP. A class of
compounds known as PDE inhibitors has been extensively studied for
use in treatment of inflammatory diseases, such as asthma, COPD and
acute respiratory distress syndrome. Preferred are inhibitors of
PDE type 1, 2, 3, 4, 7, 8, 10 or 11; in one aspect this includes
cAMP-PDE inhibitors that are selective PDE type 4 inhibitors or
inhibitors having selectivity for one particular type of PDE 4
isoenzyme, such as, by way of example, rolipram, cilomilast,
ibudilast, and piclamilast. In general, the methods and
compositions of this invention may comprise use of one or more
cAMP-PDE inhibitors.
7. Representative Compounds of the Invention
[0226] The invention is further illustrated by the following
non-limiting examples of representative compounds.
1.
N-[(1R)-2-[4-[(1-acetyl-4-piperidyl)-isobutyl-amino]piperazin-1-yl]-1-[-
(4-chlorophenyl)methyl]-2-oxo-ethyl]-2-amino-2-methyl-propanamide
[0227] The compound was synthesized using the methods of the
foregoing synthetic schemes. The compound had the structural
formula:
##STR00047##
and a molecular formula of C.sub.28H.sub.45ClN.sub.6O.sub.3. The
compound was prepared as the TFA salt, and had a molecular weight
of 549.15 without salt, and 777.19 with TFA salt.
[0228] In binding studies, the compound had a Ki of 5 nM at MC4r
and 200 nM at MC1r. In functional studies in a human MC4r
expression system approximating maximal physiologic receptor
densities, the compound had a relative efficacy, compared to
NDP-.alpha.-MSH, of 33%, with an EC.sub.50 value of 23 nM.
2.
N-[(1R)-2-[4-[(1-acetyl-4-piperidyl)-isobutyl-amino]piperazin-1-yl]-1-[-
(4-chlorophenyl)methyl]-2-oxo-ethyl]-1-amino-cyclopropanecarboxamide
[0229] The compound was synthesized using the methods of the
foregoing synthetic schemes. The compound had the structural
formula:
##STR00048##
and a molecular formula of C.sub.28H.sub.43ClN.sub.6O.sub.3. The
compound was prepared as the TFA salt, and had a molecular weight
of 547.13 without salt, and 775.17 with TFA salt.
[0230] In binding studies, the compound had a Ki of 5 nM at MC4r
and 40 nM at MC1r. In functional studies in a human MC4r expression
system approximating maximal physiologic receptor densities, the
compound had a relative efficacy, compared to NDP-.alpha.-MSH, of
3%.
3.
N-[(1R)-2-[4-[(1-acetyl-4-piperidyl)-isobutyl-amino]piperazin-1-yl]-1-[-
(4-chlorophenyl)methyl]-2-oxo-ethyl]-1-amino-cyclobutanecarboxamide
[0231] The compound was synthesized using the methods of the
foregoing synthetic schemes. The compound had the structural
formula:
##STR00049##
and a molecular formula of C.sub.29H.sub.45ClN.sub.6O.sub.3. The
compound was prepared as the TFA salt, and had a molecular weight
of 561.16 without salt, and 789.20 with TFA salt.
[0232] In binding studies, the compound had a Ki of 3 nM at MC4r.
In functional studies in a human MC4r expression system
approximating maximal physiologic receptor densities, the compound
had a relative efficacy, compared to NDP-.alpha.-MSH, of 10%.
4.
N-[(1R)-2-[4-[(1-acetyl-4-piperidyl)-isobutyl-amino]piperazin-1-yl]-1-[-
(4-chlorophenyl)methyl]-2-oxo-ethyl]-1-amino-cyclopentanecarboxamide
[0233] The compound was synthesized using the methods of the
foregoing synthetic schemes. The compound had the structural
formula:
##STR00050##
and a molecular formula of C.sub.30H.sub.47ClN.sub.6O.sub.3. The
compound was prepared as the TFA salt, and had a molecular weight
of 575.19 without salt, and 803.23 with TFA salt.
[0234] In binding studies, the compound had a Ki of 4 nM at MC4r.
In functional studies in a human MC4r expression system
approximating maximal physiologic receptor densities, the compound
had a relative efficacy, compared to NDP-.alpha.-MSH, of 10%.
5.
N-[(1R)-2-[4-[(1-acetyl-4-piperidyl)-isobutyl-amino]piperazin-1-yl]-1-[-
(4-chlorophenyl)methyl]-2-oxo-ethyl]-1-amino-cyclohexanecarboxamide
[0235] The compound was synthesized using the methods of the
foregoing synthetic schemes. The compound had the structural
formula:
##STR00051##
and a molecular formula of C.sub.31H.sub.49ClN.sub.6O.sub.3. The
compound was prepared as the TFA salt, and had a molecular weight
of 589.21 without salt, and 817.25 with TFA salt.
[0236] In binding studies, the compound had a Ki of 4 nM at MC4r.
In functional studies in a human MC4r expression system
approximating maximal physiologic receptor densities, the compound
had a relative efficacy, compared to NDP-.alpha.-MSH, of 15%, with
an EC.sub.50 value of 9 nM.
6.
N-[(1R)-2-[4-[(1-acetyl-4-piperidyl)-isobutyl-amino]piperazin-1-yl]-1-[-
(4-chlorophenyl)methyl]-2-oxo-ethyl]-2-amino-2-ethyl-butanamide
[0237] The compound was synthesized using the methods of the
foregoing synthetic schemes. The compound had the structural
formula:
##STR00052##
and a molecular formula of C.sub.30H.sub.49ClN.sub.6O.sub.3. The
compound was prepared as the TFA salt, and had a molecular weight
of 577.20 without salt, and 805.24 with TFA salt.
[0238] In binding studies, the compound had a Ki of 13 nM at MC4r.
In functional studies in a human MC4r expression system
approximating maximal physiologic receptor densities, the compound
had a relative efficacy, compared to NDP-.alpha.-MSH, of 34%, with
an EC.sub.50 value of 25 nM.
7.
N-[(1R)-2-[4-[(1-acetyl-4-piperidyl)-isobutyl-amino]piperazin-1-yl]-1-[-
(4-chlorophenyl)methyl]-2-oxo-ethyl]-1-amino-cycloheptanecarboxamide
[0239] The compound was synthesized using the methods of the
foregoing synthetic schemes. The compound had the structural
formula:
##STR00053##
and a molecular formula of C.sub.32H.sub.51ClN.sub.6O.sub.3. The
compound was prepared as the TFA salt, and had a molecular weight
of 603.24 without salt, and 831.28 with TFA salt.
[0240] In binding studies, the compound had a Ki of 3 nM at MC4r.
In functional studies in a human MC4r expression system
approximating maximal physiologic receptor densities, the compound
had a relative efficacy, compared to NDP-.alpha.-MSH, of 8%.
8.
4-[[4-[(2R)-2-[(2-amino-2-methyl-propanoyl)amino]-3-(4-chlorophenyl)pro-
panoyl]piperazin-1-yl]-isobutyl-amino]-N-ethyl-piperidine-1-carboxamide
[0241] The compound was synthesized using the methods of the
foregoing synthetic schemes. The compound had the structural
formula:
##STR00054##
and a molecular formula of C.sub.29H.sub.48ClN.sub.7O.sub.3. The
compound was prepared as the TFA salt, and had a molecular weight
of 578.19 without salt, and 806.23 with TFA salt.
[0242] In binding studies, the compound had a Ki of 6 nM at MC4r
and 150 nM at MC1r. In functional studies in a human MC4r
expression system approximating maximal physiologic receptor
densities, the compound had a relative efficacy, compared to
NDP-.alpha.-MSH, of 28%, with an EC.sub.50 value of 52 nM.
9.
N-[(1R)-2-[4-[(1-acetyl-4-piperidyl)-isobutyl-amino]piperazin-1-yl]-1-[-
(4-chlorophenyl)methyl]-2-oxo-ethyl]-2-methyl-2-(methylamino)propanamide
[0243] The compound was synthesized using the methods of the
foregoing synthetic schemes. The compound had the structural
formula:
##STR00055##
and a molecular formula of C.sub.29H.sub.47ClN.sub.6O.sub.3. The
compound was prepared as the TFA salt, and had a molecular weight
of 563.17 without salt, and 677.19 with TFA salt.
[0244] In binding studies, the compound had a Ki of 5 nM at MC4r.
In functional studies in a human MC4r expression system
approximating maximal physiologic receptor densities, the compound
had a relative efficacy, compared to NDP-.alpha.-MSH, of 19%, with
an EC.sub.50 value of 5 nM.
10.
2-amino-N-[(1R)-1-[(4-chlorophenyl)methyl]-2-[4-[[1-(2-hydroxyacetyl)--
4-piperidyl]-isobutyl-amino]piperazin-1-yl]-2-oxo-ethyl]-2-methyl-propanam-
ide
[0245] The compound was synthesized using the methods of the
foregoing synthetic schemes. The compound had the structural
formula:
##STR00056##
and a molecular formula of C.sub.28H.sub.45ClN.sub.6O.sub.4. The
compound was prepared as the TFA salt, and had a molecular weight
of 565.15 without salt, and 793.19 with TFA salt.
[0246] In binding studies, the compound had a Ki of 5 nM at MC4r.
In functional studies in a human MC4r expression system
approximating maximal physiologic receptor densities, the compound
had a relative efficacy, compared to NDP-.alpha.-MSH, of 22%, with
an EC.sub.50 value of 9 nM.
11.
N-[(1R)-2-[4-[(1-acetyl-4-piperidyl)-isobutyl-amino]piperazin-1-yl]-1--
[(4-chlorophenyl)methyl]-2-oxo-ethyl]-2-methyl-pyrrolidine-2-carboxamide
[0247] The compound was synthesized using the methods of the
foregoing synthetic schemes. The compound had the structural
formula:
##STR00057##
and a molecular formula of C.sub.30H.sub.47ClN.sub.6O.sub.3. The
compound was prepared as the TFA salt, and had a molecular weight
of 575.19 without salt, and 803.23 with TFA salt.
[0248] In binding studies, the compound had a Ki of 6 nM at MC4r.
In functional studies in a human MC4r expression system
approximating maximal physiologic receptor densities, the compound
had a relative efficacy, compared to NDP-.alpha.-MSH, of 13%, with
an EC.sub.50 value of 6 nM.
8. Examples
[0249] The invention is further illustrationed by the following
non-limiting examples.
[0250] 1. Receptor Binding and Function
[0251] Melanocortin receptor binding and function of Compound 7.1
was tested. Ki binding values (nM) were calculated utilizing the
percent inhibit of the binding of iodine-125 labeled
NDP-.alpha.-MSH against B16-F1 cells for mMC1r, and human
recombinant CHO cells for hMC3r, hMC4r and hMC5r, with the
inhibition constant calculated using the Cheng Prusoff equation.
Functional results (EC.sub.50 and efficacy) were calculated
utilizing cAMP assays with NDP-.alpha.-MSH for mMC1r, hMC3r and
hMC4r, ACTH for hMC2r, and .alpha.-MSH for hMC5r for agonist
measures, with the same target cells for melanocortin receptors
mMC1r, hMC3r, hMC4r and hMC5r, and human recombinant Cloudman M3
cells for hMC2r.
TABLE-US-00003 Melanocortin Receptor Binding and Function of
Compound 7.1 cAMP cAMP Binding, EC.sub.50 (nM) Efficacy (%) Ki (nM)
mMC1r 460 72 93 hMC2r >1000 22 -- hMC3r >1000 28 1300 hMC4r 6
91 14 hMC5r >1000 20 1200
[0252] 2. Body Weight and Food Intake
[0253] Compound 7.1 was assessed in a body weight and food intake
in vivo study in diet induced obese (010) mice. C57BL/6 male DIO
mice were utilized, that were 17 weeks of age on arrival. Animals
were pre-conditioned on a high-fat diet for 11 weeks by Jackson
Labs. Weight variation of animals at the time of treatment
initiation did not exceed .+-.20% of the mean weight. Animals were
single housed within a limited access rodent facility, and provided
ad libitum with a commercial rodent diet and free access to
drinking water. Animals were randomized into five groups, a vehicle
group, three dose levels of compound 7.1 (3 mg/kg, 10 mg/kg and 30
mg/kg), and Peptide A, a positive control cyclic peptide of the
formula Ac-Arg-cyclo(Orn-Asn-D-Phe-Arg-Trp-Glu)-NH.sub.2. Following
three days of sham dosing, animals were dosed for three days, with
vehicle or Compound 7.1 dosed orally once daily at within one hour
of lights out in the AM, and Peptide A dosed by subcutaneous
injection twice daily within one hour before lights out and six
hours after lights out. Body weights were measured one hour prior
to lights out, and feed weights were measured both one hour prior
to lights out and six hours post lights out.
TABLE-US-00004 Mean .+-. SEM Group Body Weight Change Body Weight
Change (%) Day 0 Day 1 Day 2 Day 3 Treatment Mean SEM N Mean SEM N
Mean SEM N Mean SEM N Vehicle 0 0 9 -0.68 0.32 9 -1.80 0.42 9 -1.99
0.44 9 7.1 (3 mg/kg) 0 0 9 -1.97 0.52 9 -2.92 0.69 9 -3.80* 0.81 9
7.1 (10 mg/kg) 0 0 9 -2.48* 0.21 9 -3.77** 0.41 9 -4.35** 0.47 9
7.1 (30 mg/kg) 0 0 9 -4.41** 0.42 9 -5.82** 0.48 9 -7.08** 0.79 9
Peptide A (1 mg/kg) 0 0 9 -5.69** 0.39 9 -8.37** 0.38 9 -8.78**
0.54 9
TABLE-US-00005 Mean .+-. SEM Group Feed Intake at 24 Hours Post
Lights Out Food Intake - 24 Hours (g) Day 0 Day 1 Day 2 Treatment
Mean SEM N Mean SEM N Mean SEM N Vehicle 2.23 0.19 9 2.23 0.12 9
2.47 0.08 9 7.1 1.90 0.13 9 2.03 0.17 9 2.11 0.15 9 (3 mg/kg) 7.1
1.82 0.08 9 1.78 0.14 9 2.20 0.19 9 (10 mg/kg) 7.1 1.62* 0.12 9
1.60* 0.20 9 1.91* 0.16 9 (30 mg/kg) Peptide A 0.99** 0.10 9 1.10**
0.14 9 1.67** 0.18 9 (1 mg/kg)
[0254] *p<0.05 vs. sham (vehicle control) using two-way ANOVA
followed by Dunnett's post-test.
[0255] **p<0.01 vs sham (vehicle control) using two-way ANOVA
followed by Dunnett's post-test.
[0256] Daily treatment with compound 7.1 at 10 mg/kg and 30 mg/kg
resulted in significant body weight loss from day 1 to 3. Treatment
with 3 mg/kg of compound 7.1 resulted in body weight loss on day 3.
Two times daily treatment with 1 mg/kg of Peptide A decreased
animal body weights throughout the study. At 24 hours post lights
out, treatment with 30 mg/kg compound 7.1 or 1 mg/kg Peptide A
significantly inhibited feed intake by animals through the entirety
of the study, compared with sham animals.
[0257] 3. Plasma Concentrations Following Oral Administration
[0258] As shown in FIG. 1, plasma concentrations of the compound
7.1 were determined in Sprague-Dawley rats following oral
administration of 30 mg/kg of compound 7.1. Plasma samples were
measured at 15 minutes (2 animals) and 30 minutes, 1 hour, 2 hours,
4 hours, 8 hours and 24 hours (3 animals) post oral administration.
The terminal half-life was 3.94 hours following the dose of 30
mg/kg, with a Tax of 2 hours and Cmax of 780 ng/mL.
[0259] 4. Plasma Concentration at One Hour Following Oral
Administration of Three Dose Levels
[0260] In the study of Example 8.2, at one hour after
administration of 3 mg/kg, 10 mg/kg or 30 mg/kg of compound 7.1, a
terminal blood draw was conducted in the DIO mice, and the plasma
samples were measure for concentration of compound 7.1. Results are
shown in FIG. 2.
[0261] 5. Oral Administration in Leptin-Deficient Obese Mice
[0262] Male mice homozygous for the obese spontaneous mutation,
Lep.sup.ob, with resulting leptin protein deficiency, were
individually housed with free access to feed and drinking water.
All animals were sham-dosed twice daily for five days prior to
initial drug administration, utilizing a vehicle dose of 3.2%
mannitol and 55 nM tris(hydroxymethyl)aminomethane at pH 7.4. On
day 6 a cohort of the mice received compound 7.1 dissolved into the
vehicle twice daily for six consecutive days at a dose of 30 mg/kg.
Body weight of animals was measured daily, with data analyzed as
the percent change from baseline of each individual animal, where
baseline was defined as the average weight during the first three
days of sham dosing. The averaged percent change from baseline of a
vehicle group was then subtracted from each value to correct for
continued weight gain or other study-wide factors. The change in
body weight is shown in FIG. 3 starting on study day 1, the first
day of administration of compound 7.1. Leptin-deficient mice
treated with compound 7.1 showed a 4.8% decrease in body weight of
4.8% compared to vehicle controls at a dose of 30 mg/kg of compound
7.1 administered twice daily.
[0263] 6. Oral Administration in MC4r Knockout Mice
[0264] Feed intake and body weight was evaluated in MCr4 knockout
(MC4r-/-) mice (Jackson Laboratories) compared to C57Bl/6 DIO mice.
Two daily administration of 30/mg/kg of compound 7.1 via oral
gavage resulted in significant body weight loss in DIO mice, but
not in MC4r (-/-) mice, when each cohort was compared to their own
vehicle group. Feed weights were taken at time 0, 2, 6 and 24 hours
post lights out. There were no differences in food consumption in
the MC4r (-/-) mice receiving compound 7.1 compared to MC4r (-/-)
mice receiving vehicle only, but in the DIO mice food consumption
was significantly decreased at 6 and 24 hour timepoints in mice
receiving compound 7.1 compared to DIO mice receiving vehicle only.
The change in body weight is shown in FIG. 4 starting on study day
5, the first day of administration of compound 7.1. There was a
notable mean percentage body weight change difference (4.9%, 95% Cl
(3.237, 6.538)) between the treated group and vehicle for DIO mice
but not for MC4r (-/-) mice. Additionally, there was a
statistically significant mean difference, 2.8%, 95% Cl (1.281,
4.253), and p value <0.0001 between DIO mice receiving compound
7.1 and MC4r (-/-) receiving compound 7.1.
[0265] Each of the foregoing is merely illustrative, and other
equivalent embodiments are possible and contemplated.
[0266] Although this invention has been described with reference to
these preferred embodiments, other embodiments can achieve the same
results. Variations and modifications of this invention will be
obvious to those skilled in the art and it is intended to cover in
the appended claims all such modifications and equivalents. The
entire disclosures of all applications, patents, and publications
cited above are hereby incorporated by reference.
[0267] The preceding examples can be repeated with similar success
by substituting the generically or specifically described reactants
and/or operating conditions of this invention for those used in the
preceding examples.
* * * * *