U.S. patent application number 16/878097 was filed with the patent office on 2020-09-03 for therapies for obesity, diabetes and related indications.
The applicant listed for this patent is Palatin Technologies, Inc.. Invention is credited to John H. Dodd, Marie Makhlina, Carl Spana.
Application Number | 20200276275 16/878097 |
Document ID | / |
Family ID | 1000004838657 |
Filed Date | 2020-09-03 |
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United States Patent
Application |
20200276275 |
Kind Code |
A1 |
Spana; Carl ; et
al. |
September 3, 2020 |
Therapies for Obesity, Diabetes and Related Indications
Abstract
The invention relates to methods, uses, compositions and
formulations including a melanocortin receptor-4 agonist and a
glucagon-like peptide-1 receptor agonist for treatment of obesity,
diabetes, metabolic syndrome and related indications, diseases or
disorders.
Inventors: |
Spana; Carl; (West Harrison,
NY) ; Dodd; John H.; (Spring Mils, PA) ;
Makhlina; Marie; (Highland Park, NJ) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Palatin Technologies, Inc. |
Cranbury |
NJ |
US |
|
|
Family ID: |
1000004838657 |
Appl. No.: |
16/878097 |
Filed: |
May 19, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15723755 |
Oct 3, 2017 |
10660939 |
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16878097 |
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PCT/US2016/027402 |
Apr 14, 2016 |
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15723755 |
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62147256 |
Apr 14, 2015 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 3/00 20180101; A61K
9/0019 20130101; A61K 38/12 20130101; A61K 38/26 20130101; A61P
3/04 20180101; A61P 3/10 20180101 |
International
Class: |
A61K 38/26 20060101
A61K038/26; A61K 9/00 20060101 A61K009/00; A61K 38/12 20060101
A61K038/12; A61P 3/00 20060101 A61P003/00; A61P 3/04 20060101
A61P003/04; A61P 3/10 20060101 A61P003/10 |
Claims
1. A method of decreasing side effects associated with therapeutic
agents for treatment of obesity, diabetes or metabolic syndrome in
a patient, comprising: administration of a quantity of melanocortin
receptor-4 agonist, wherein the quantity of melanocortin receptor-4
agonist administered, if administered as a monotherapy not in
conjunction with glucagon-like peptide-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 administration of a quantity of glucagon-like
peptide-1 receptor agonist, wherein the quantity of glucagon-like
peptide-1 receptor agonist administered, if administered as a
monotherapy not in conjunction with melanocortin receptor-4
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; wherein the quantity of the
melanocortin receptor-4 agonist and the quantity of glucagon-like
peptide-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, thereby reducing side effects in the treatment of
at least one of obesity, diabetes or metabolic syndrome in the
patient.
2. The method of claim 1 wherein the quantity of melanocortin
receptor-4 agonist administered is not sufficient to initiate the
desired pharmacological response of inducing weight loss.
3. The method of claim 1 or 2 wherein the quantity of melanocortin
receptor-4 agonist administered is sufficient to induce minimal
weight loss.
4. The method of claim 1 wherein the desired pharmacological
response is inducing weight loss.
5. The method of claim 1 wherein the desired pharmacological
response is inducing glycemic control.
6. A method of treating obesity or inducing weight loss in an obese
patient, comprising the steps of: establishing a dose of a
glucagon-like peptide-1 receptor agonist which induces glycemic
control in the patient but which induces no more than minimal
weight loss in the patient; administration of the dose of the
glucagon-like peptide-1 receptor agonist; and administration of a
melanocortin receptor-4 agonist; wherein the dose of the
glucagon-like peptide-1 receptor agonist and the quantity of
melanocortin receptor-4 agonist are effective to treat obesity or
to induce weight loss.
7. The method of claim 6 wherein the dose of glucagon-like
peptide-1 receptor agonist does not induce minimal weight loss in
the patient.
8. The method of claim 6 wherein the administration of the dose of
the glucagon-like peptide-1 receptor agonist and administration of
a melanocortin receptor-4 agonist are simultaneous.
9. The method of claim 8 wherein the simultaneous administration is
by means of a combination pharmaceutical composition comprising
glucagon-like peptide-1 receptor agonist and melanocortin
receptor-4 agonist.
10. The method of claim 6 wherein the melanocortin receptor-4
agonist is Ac-Arg-cyclo(Orn-Asn-D-Phe-Arg-Trp-Glu)-NH.sub.2
administered daily.
11. The method of claim 6 wherein
Ac-Arg-cyclo(Orn-Asn-D-Phe-Arg-Trp-Glu)-NH.sub.2 is administered by
subcutaneous injection.
12. The method of claim 6 wherein the glucagon-like peptide-1
receptor agonist is administered by subcutaneous injection.
13. The method of claim 12 wherein the glucagon-like peptide-1
receptor agonist is liraglutide or exenatide administered daily or
twice daily.
14. The method of claim 12 wherein the glucagon-like peptide-1
receptor agonist is lixisenatide, albiglutide, dulaglutide or an
extended release formulation of exenatide or liraglutide
administered at weekly or greater intervals.
15. The method of claim 6 wherein administration of the dose of the
glucagon-like peptide-1 receptor agonist and administration of a
melanocortin receptor-4 agonist elicits a synergistic effect on
treatment of obesity.
16. The method of claim 6 wherein administration of the dose of the
glucagon-like peptide-1 receptor agonist and administration of a
melanocortin receptor-4 agonist elicits a synergistic effect on
inducing glycemic control.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a continuation application of U.S.
patent application Ser. No. 15/723,755, entitled "Therapies for
Obesity, Diabetes and Related Indications", filed on Oct. 3, 2017,
and now issued as U.S. Pat. No. 10,660,939 on May 26, 2020, which
in turn is a continuation of International Application No.
PCT/US2016/027402, published as International Publication No. WO
2016/168388, entitled "Therapies for Obesity, Diabetes and Related
Indications", filed Apr. 14, 2016, which in turn claims priority to
and the benefit of the filing of U.S. Provisional Patent
Application Ser. No. 62/147,256 entitled "Therapies for Obesity,
Diabetes and Related Indications", filed Apr. 14, 2015, and the
specification and claims thereof are incorporated herein by
reference.
BACKGROUND OF THE INVENTION
Field of the Invention (Technical Field)
[0002] The present invention relates to medical use of compounds
specific for melanocortin receptors and compounds specific for
glucagon-like peptide-1 receptors, for example in the treatment of
obesity, diabetes, metabolic syndrome and related indications,
diseases or disorders.
Description of Related Art
[0003] The following discussion refers to a number of publications
by author(s) and year of publication, and that due to recent
publication dates certain publications are not to be considered as
prior art vis-a-vis the present invention. Discussion of such
publications herein is given for more complete background and is
not to be construed as an admission that such publications are
prior art for patentability determination purposes.
[0004] 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 (MCSr), 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 MCSr is believed to be involved in exocrine gland
system regulation and other functions.
[0005] 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.
[0006] 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)).
Agouti-related protein (AgRP) is an endogenous compound that has
been suggested to be an antagonist or an inverse agonist on MC4r.
.alpha.-MSH is believed to be the principle endogenous MC4r
agonist.
[0007] Peptides specific for MC4r, including MC4r agonists, and
secondarily peptides specific for MC3r, are believed to be useful
in regulation of mammalian energy homeostasis, including use as
agents for attenuating food intake and body weight gain.
[0008] Melanocortin receptor-specific cyclic peptides include
.alpha.-MSH analog peptides such as
Ac-Nle-cyclo(-Asp-His-D-Phe-Arg-Trp-Lys)-NH.sub.2 (See U.S. Pat.
Nos. 5,674,839 and 5,576,290) and
Ac-Nle-cyclo(-Asp-His-D-Phe-Arg-Trp-Lys)-OH (See U.S. Pat. Nos.
6,579,968 and 6,794,489). These and other melanocortin
receptor-specific peptides generally contain the central
tetrapeptide sequence of native .alpha.-MSH,
His.sup.6-Phe.sup.7-Arg.sup.8-Trp.sup.9, or a mimetic or variation
thereof, such as the substitution of D-Phe for Phe'. Other
melanocortin receptor-specific cyclic peptides, including primarily
MC4r specific peptides, are disclosed in U.S. Pat. Nos. 7,326,707;
7,354,923; 7,456,184; 7,834,017; 7,964,601; 7,968,548; 8,455,617;
8,455,618; 8,487,073; 8,492,517; 8,729,224; 8,846,601; 8,877,890;
and 8,933,194; and applications US 2012/0077957 and US
2014/0357575. Melanocortin receptor-specific cyclic peptides
disclosed in the foregoing are typically cyclized through a lactam
bridge, or alternatively through a disulfide bridge formed by the
side chains of two Cys or other reactive thiol-containing
residues.
[0009] Also known are 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). 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.
[0010] 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-N H2 (SEQ ID NO:1)
(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.
[0011] 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.
[0012] While both GLP-1 receptor agonists and MC4r agonists have
utility in treatment of obesity, diabetes mellitus type 2,
metabolic syndrome and other conditions, there are limitations on
the utility of each, particularly for treatment of obesity,
metabolic syndrome and related indications. It is against this
background that the present invention was made.
BRIEF SUMMARY OF THE INVENTION
[0013] The invention provides a pharmaceutical composition for
subcutaneous administration in treatment of obesity or to induce
weight loss, comprising on a per dose basis:
[0014] a MC4r agonist 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
[0015] 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 a MC4r agonist. The
pharmaceutical composition preferably has a synergistic
anti-obesity effect. In the composition formulated for once daily
administration, the MC4r agonist may be Peptide No. 154 and the
GLP-1 receptor agonist may be liraglutide, preferable between about
1.5 mg and 10 mg of Peptide No. 154 and between about 0.6 mg and
1.5 mg of liraglutide on a per dose basis. In the composition
formulated for once daily administration, the MC4r agonist may be
Peptide No. 154 and the GLP-1 receptor agonist may be exenatide,
preferably between about 0.75 mg and 5 mg of Peptide No. 154 and
between about 5 .mu.g and 20 .mu.g of exenatide on a per dose
basis. The composition may further include either or both a
pharmaceutically acceptable salt and a pharmaceutically acceptable
carrier.
[0016] In another aspect, the invention provides a pharmaceutical
composition comprising as components: [0017] (a) a MC4r agonist,
and [0018] (b) a GLP-1 receptor agonist;
[0019] wherein components (a) and (b) are present in such weight or
molar ratio that the composition exerts a synergistic effect in
treatment of obesity or to induce weight loss upon administration
to a patient. This composition may include at least one
pharmaceutically acceptable carrier. If formulated for once daily
administration, the MC4r agonist may be Peptide No. 154 and the
GLP-1 receptor agonist may be liraglutide, such as between about
1.5 mg and 10 mg of Peptide No. 154 and between about 0.6 mg and
1.5 mg of liraglutide on a per dose basis. If formulated for twice
daily administration, the composition may include between about
0.75 mg and 5 mg of Peptide No. 154 and between about 5 .mu.g and
20 .mu.g of exenatide on a per dose basis.
[0020] In another aspect, the invention provides a method of
treating a patient with obesity, diabetes or metabolic syndrome,
comprising administering to the patient (a) a MC4r agonist 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 a MC4r
agonist. Preferably the method elicits a synergistic effect on
treatment of obesity. The quantity and schedule of administration
of the MC4r agonist 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 MC4r agonist and the GLP-1
receptor agonist may together be sufficient to produce synergistic
effect in the treatment of glycemic control. In the method, the
MC4r agonist may be Peptide No. 154 administered daily by
subcutaneous injection 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. The MC4r agonist and the GLP-1
receptor agonist may be administered simultaneously to the patient,
sequentially to the patient in either order or via different
pathways of administration. In the method, either or both the MC4r
agonist and the GLP-1 may be sustained, 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. In a related aspect, the MC4r agonist is not
sustained-release.
[0021] 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:
[0022] administration of a quantity of MC4r agonist, wherein the
quantity of MC4r agonist 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
[0023] 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 MC4r 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;
[0024] wherein the quantity of the MC4r agonist 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,
[0025] thereby reducing side effects in the treatment of at least
one of obesity, diabetes or metabolic syndrome in the patient. 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. The desired
pharmacological response may be inducing weight loss or inducing
glycemic control.
[0026] In another aspect, the invention provides a method of
treating obesity or inducing weight loss in an obese patient,
comprising the steps of:
[0027] 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;
[0028] administration of the dose of the GLP-1 receptor agonist;
and
[0029] administration of a MC4r agonist;
[0030] wherein the dose of the GLP-1 receptor agonist and the
quantity of MC4r agonist are effective to treat obesity or to
induce weight loss.
In this method, the dose of GLP-1 receptor agonist preferably does
not induce minimal weight loss in the patient. The administration
of the dose of the GLP-1 receptor agonist and administration of a
MC4r agonist may be simultaneous, such as by means of a combination
pharmaceutical composition comprising GLP-1 receptor agonist and
MC4r agonist. In the practice of the method, the MC4r agonist may
be Peptide No. 154 administered daily, such as by subcutaneous
injection. The GLP-1 receptor agonist may also 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.
[0031] In another aspect, the present invention provides a
combination therapeutic pharmaceutical composition for use in the
treatment of obesity, diabetes and related indications, comprising
a MC4r agonist and a GLP-1 receptor agonist.
[0032] In another aspect, the present invention provides
combination therapy comprising a peptide-based melanocortin
receptor-specific pharmaceutical, wherein the peptide is a
selective MC4r agonist, together with a GLP-1 receptor agonist.
[0033] Other aspects and novel features, and the further scope of
applicability of the present 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 the invention. The
aspects of the 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
[0034] 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.
[0035] FIG. 1 depicts percent change in the area under the curve
(AUC) for a glucose tolerance test from the baseline test to the
termination test for diet induced obese (DIO) mice administered
vehicle, an MC4r agonist (Peptide No. 154, hereafter "MC4r 154"),
combinations of MC4r 154 and GLP-1, or GLP-1 with a caloric
restriction to mimic food intake with MC4r 154, all .+-.standard
deviation (SD) from the baseline test. All compounds were
administered via subcutaneous osmotic pump on study days 0 to 5.
For combination therapy the agents were co-formulated into one
solution and administered via a single osmotic pump. For the GLP-1
caloric restriction group the pump implantation was delayed by one
day and the amount of food given to the animals was matched to
those receiving MC4r 154 alone. The glucose tolerance test was
conducted on day -2 (baseline) and day 6 (termination) by
administering glucose orally at 1 g/kg and measuring blood glucose
levels at various time points.
[0036] FIG. 2 depicts average body weight change in grams in DIO
mice from study day 0 through study day 5 in DIO mice administered
vehicle, MC4r 154, combinations of MC4r 154 and GLP-1, or GLP-1
combined with a caloric restriction to mimic food intake with MC4r
154, .+-.standard error of the mean (SEM). All compounds were
administered via subcutaneous osmotic pump on study days 0 to 5.
For combination therapy the agents were co-formulated into one
solution and administered via a single osmotic pump. For the GLP-1
caloric restriction group the pump implantation was delayed by one
day and the amount of food given to the animals was matched to
those receiving MC4r 154 alone. Body weight was measured daily
prior to beginning of the feeding cycle.
[0037] FIG. 3 depicts percent change in glucose from the baseline
test (study day -2) to the termination test (study day 12) in DIO
Sprague-Dawley rats, .+-.SEM, with animals receiving vehicle, MC4r
154, GLP-1 or combinations of MC4r 154 and GLP-1 as depicted. GLP-1
was administered via subcutaneously implanted osmotic pump on study
days 0 to 12. MC4r 154 was administered twice daily via
subcutaneous injection on study days 7 through 12.
[0038] FIG. 4 depicts fasted insulin levels from the baseline test
(study day -2) to termination test (study day 12) in ng/mL insulin
.+-.SEM, with animals receiving vehicle, MC4r 154, GLP-1 or
combinations of MC4r 154 and GLP-1 as depicted. GLP-1 was
administered via subcutaneously implanted osmotic pump on study
days 0 to 12. MC4r 154 was administered twice daily via
subcutaneous injection on study days 7 through 12.
[0039] FIG. 5 depicts percent change in the AUC for a glucose
tolerance test from the baseline test (study day -2) to the
termination test (study day 10) for DIO rats administered vehicle,
an MC4r agonist (MC4r 154), or a combination of MC4r agonist and
GLP-1. GLP-1 was administered via subcutaneously implanted osmotic
pump on study days 0 to 10. MC4r agonist was administered twice
daily via subcutaneous injection on study days 5 through 10.
[0040] FIGS. 6 through 8 depict average body weight change in
percent of baseline in DIO rats from study day 0 through study day
9 in DIO rats administered vehicle, MC4r 154, or a combination of
MC4r 154 and GLP-1 on study day 5, at the doses depicted in FIGS. 6
through 8. GLP-1 was administered via subcutaneously implanted
osmotic pump on study days 0 to 10. MC4r 154 was administered twice
daily via subcutaneous injection on study days 5 through 10. Body
weight was measured daily prior to the beginning of the feeding
cycle and prior to the first dose of MC4r 154.
DETAILED DESCRIPTION OF THE INVENTION
Definitions
[0041] Before proceeding with the description of the invention,
certain terms are defined as set forth herein.
[0042] In the sequences given for the peptides according to the
present invention, the amino acid residues have their conventional
meaning as given in Chapter 2400 of the Manual of Patent Examining
Procedure 9th Ed. Thus, "Ala" is alanine, "Asn" is asparagine,
"Asp" is aspartic acid, "Arg" is arginine, "Cys" is cysteine, "Gly"
is glycine, "Gln" is glutamine, "Glu" is glutamic acid, "His" is
histidine, "Ile" is isoleucine, "Leu" is leucine, "Lys" is lysine,
"Met" is methionine, "Phe" is phenylalanine, "Pro" is proline,
"Ser" is serine, "Thr" is Threonine, "Trp" is tryptophan, "Tyr" is
tyrosine, and "Val" is valine, and so on. It is to be understood
that "D" isomers are designated by a "D-" before the three letter
code or amino acid name, such that for example D-Phe is
D-phenylalanine. Amino acid residues not encompassed by the
foregoing have the following definitions:
TABLE-US-00001 Abbreviation Common Name Side Chain or Amino Acid
Structure Aib alpha-aminoisobutyric acid ##STR00001## Aic
2-aminoindane-2- carboxylic acid ##STR00002## Cit citrulline
##STR00003## Dab diaminobutyric acid ##STR00004## Dab(Acetyl)
2-amino, 4- acetylaminobutyric acid ##STR00005## Dab(Glycyl)
2-amino, 4- (glycyl)aminobutyric acid ##STR00006## Dap
diaminoproprionic acid ##STR00007## hGlu homoglutamic acid
##STR00008## Hyp hydroxyproline ##STR00009## Hyp(Bzl) O-benzyl-
hydroxyproline ##STR00010## Met(O.sub.2) Methionine sulfone
##STR00011## Met(=O) Methionine sulfoxide ##STR00012## Nal 1
3-(1-naphthyl)alanine ##STR00013## Nal 2 3-(2-naphthyl)alanine
##STR00014## Nle norleucine ##STR00015## Orn ornithine ##STR00016##
Phe(2-C(=O)- NH.sub.2+L) 2-carbamoyl- phenylalanine ##STR00017##
Phe(3-C(=O)- NH.sub.2) 3-carbamoyl- phenylalanine ##STR00018##
Phe(4-C(=O)- NH.sub.2+L) 4-carbamoyl- phenylalanine ##STR00019##
Pro(4R-Bzl) 4(R)benzyl-proline ##STR00020## Pro(4R-NH.sub.2)
4(R)amino-proline ##STR00021## Sar sarcosine ##STR00022## Ser(Bzl)
O-benzyl-serine ##STR00023## Thr(BzI) O-benzyl-threonine
##STR00024##
[0043] By a melanocortin receptor "agonist" is meant an endogenous
substance, drug substance or compound, including a compound such as
the MC4r peptides disclosed herein, which can interact with a
melanocortin receptor and initiate a pharmacological response,
including but not limited to adenyl cyclase activation,
characteristic of the melanocortin receptor. For the present
invention, a melanocortin receptor agonist which is an agonist at
melanocortin receptor-4 (MC4r agonist) is preferred.
[0044] By ".alpha.-MSH" is meant the peptide
Ac-Ser-Tyr-Ser-Met-Glu-His-Phe-Arg-Trp-Gly-Lys-Pro-Val-NH.sub.2
(SEQ ID NO:2) and analogs and homologs thereof, including without
limitation NDP-.alpha.-MSH.
[0045] By "binding affinity" is meant the ability of a compound or
drug to bind to its biological target, expressed herein as Ki
(nM).
[0046] The term "combination therapy" is intended to mean the use
of more than one active ingredient in treatment of any disease,
condition or syndrome. A combination therapy may be a composition
comprising more than one active ingredient, or may be the use of
different compositions, which may be administered by the same or
different routes of administration, with each composition including
at least one, including only one, active ingredient. When
combination therapy includes use of more than one composition, the
compositions may be administered at the same time or at separate
times, and thus combination therapy includes administration of
different compositions separated by a number of hours or days.
[0047] 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 utilized in the present invention
encompass any composition made by admixing an active ingredient and
one or more pharmaceutically acceptable carriers.
[0048] The term "diabetes" includes type 1 diabetes, which is
insulin-dependent diabetes mellitus as diagnosed according to
criteria published in the Report of the Expert Committee on the
Diagnosis and Classification of Diabetes Mellitus (Diabetes Care,
Vol. 24, Supp. 1, January 2001) whereby fasting plasma glucose
level is greater than or equal to 126 milligrams per deciliter and
for which the primary cause is pancreatic beta cell destruction,
type 2 diabetes, which is non-insulin-dependent diabetes mellitus
as diagnosed according to criteria published in the Report of the
Expert Committee on the Diagnosis and Classification of Diabetes
Mellitus whereby fasting plasma glucose level is greater than or
equal to 126 milligrams per deciliter, and latent autoimmune
diabetes mellitus of adults.
[0049] By "EC.sub.50" is meant the molar concentration of an
agonist, including a partial agonist, which produced 50% of the
maximum possible response for that agonist. By way of example, a
test compound which, at a concentration of 72 nM, produces 50% of
the maximum possible response for that compound as determined in a
cAMP assay in an MC4r cell expression system has an EC.sub.50 of 72
nM. Unless otherwise specified, the molar concentration associated
with an EC.sub.50 determination is in nanomoles per liter (nM).
[0050] In general, "functional activity" is a measure of the
signaling of a receptor, or measure of a change in
receptor-associated signaling, such as a melanocortin receptor, and
in particular MC4r or hMC4r, upon activation by a compound.
Melanocortin receptors initiate signal transduction through
activation of multimeric G proteins. GLP-1 receptor agonists also
initiate signal transduction through activation of multimeric G
proteins.
[0051] By "GLP-1 receptor agonist" is meant one or more naturally
occurring GLP-1 receptor agonists, including GLP-1 polypeptides
(GLP-1 (7-37)-OH and GLP-1(7-36)-NH.sub.2, GLP-1 fragments, GLP-1
analogs, GLP-1 derivatives of naturally occurring GLP-1
polypeptides, GLP-1 fragments, or GLP-1 analogs, and Exendin-3 and
Exendin-4 that have the ability to bind to the GLP-1 receptor and
initiate a signal transduction pathway resulting in insulinotropic
activity, including but not limited to exenatide, liraglutide,
lixisenatide, taspoglutide, albiglutide, and dulaglutide.
[0052] A compound is effective to "induce glycemic control" when
the compound induces, over a period of time of 12 weeks or greater,
a statistically significant and placebo-adjusted decrease in either
(a) mean hemoglobin A.sub.1c or (b) mean fasting plasma glucose of
at least about 5.0% in a cohort of subjects with a baseline mean
hemoglobin A.sub.1c of at least about 7.5% or a baseline mean
fasting plasma glucose of at least about 150 mg/dL.
[0053] By "inhibition" is meant the percent attenuation, or
decrease in receptor binding, in a competitive inhibition assay
compared to a known standard. Thus, by "inhibition at 1 .mu.M
(NDP-.alpha.-MSH)" is meant the percent decrease in binding of
NDP-.alpha.-MSH by addition of a determined amount of the compound
to be tested, such as 1 .mu.M of a test compound, such as under the
assay conditions hereafter described. By way of example, a test
compound that does not inhibit binding of NDP-.alpha.-MSH has a 0%
inhibition, and a test compound that completely inhibits binding of
NDP-.alpha.-MSH has a 100% inhibition. Typically, as described
hereafter, a radio assay is used for competitive inhibition
testing, such as with I.sup.125-labeled NDP-.alpha.-MSH, or a
lanthanide chelate fluorescent assay, such as with
Eu-NDP-.alpha.-MSH. However, other methods of testing competitive
inhibition are known, including use of label or tag systems other
than radioisotopes, and in general any method known in the art for
testing competitive inhibition may be employed in this invention.
It may thus be seen that "inhibition" is one measure to determine
whether a test compound attenuates binding of NDP-.alpha.-MSH or
.alpha.-MSH to melanocortin receptors.
[0054] By "intrinsic activity" 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 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, and a
compound with intrinsic activity less than 0.7 (70%) is classified
as a partial agonist. In one aspect, the MC4r peptides utilized in
the present invention may generally be characterized as an agonist
at MC4r with respect to .alpha.-MSH or NDP-.alpha.-MSH.
[0055] By "Ki (nM)" is meant the equilibrium inhibitor dissociation
constant representing the molar concentration of a competing
compound that binds to half the binding sites of a receptor at
equilibrium in the absence of radioligand or other competitors. In
general, the numeric value of 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):
K i = EC 50 1 + [ ligand ] K D ##EQU00001##
where "ligand" is the concentration of radioligand and K.sub.D is
an inverse measure of receptor affinity for the radioligand which
produces 50% receptor occupancy by the radioligand. Unless
otherwise specified, the molar concentration associated with a Ki
determination is in nM. Ki may be expressed in terms of specific
receptors (e.g., MC1r, MC3r, MC4r or MC5r) and specific ligands
(e.g., .alpha.-MSH or NDP-.alpha.-MSH).
[0056] The term "metabolic syndrome" refers to metabolic disorders,
particularly glucose and lipid regulatory disorders, including
insulin resistance and defective secretion of insulin by pancreatic
beta cells, and may further include conditions and states such as
abdominal obesity, dyslipidemia, hypertension, glucose intolerance
or a prothrombitic state, and which may further result in disorders
such as hyperlipidemia, obesity, diabetes, insulin resistance,
glucose intolerance, hyperglycemia, and hypertension.
[0057] A compound is effective to induce at least "minimal weight
loss" when the compound induces, over a period of time from 12 to
52 weeks, a statistically significant and placebo-adjusted decrease
in mean body weight of at least about 2.5%, but less than about
5.0%, in a cohort of subjects with a baseline mean BMI 27
kg/m.sup.2.
[0058] By "NDP-.alpha.-MSH" is meant the peptide
Ac-Ser-Tyr-Ser-Nle-Glu-His-D-Phe-Arg-Trp-Gly-Lys-Pro-Val-NH.sub.2
and analogs and homologs thereof.
[0059] The term "obesity" means the condition of excess body fat
(adipose tissue), including by way of example in accordance with
the National Institutes of Health Federal Obesity Clinical
Guidelines for adults, whereby body mass index ("BMI") calculated
by dividing body mass in kilograms by height in meters squared is
equal to or greater than twenty-five (25), and further including an
overweight condition and comparable obesity and overweight
condition in children.
[0060] The term "prophylactically effective" or "preventive" means
the amount of MC4r agonist and GLP-1 receptor agonist, including
the ratio of MC4r agonist and GLP-1 receptor agonist, which 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, including but not
limited to obesity, diabetes or exacerbation of metabolic
syndrome.
[0061] As used herein, the term "pharmacologically effective
amount" (including "therapeutically effective amount") means an
amount of the combination of MC4r agonist and GLP-1 receptor
agonist, including the ratio of MC4r agonist and GLP-1 receptor
agonist and/or the pharmacokinetic properties, including but not
limited to half-life, according to the invention that is sufficient
to induce a desired therapeutic or biological effect.
[0062] The term "sustained-release", as in a sustained-release
form, sustained-release composition or sustained-release
formulation, is intended to include a form of an active ingredient,
or formulation for an active ingredient, which has an extended in
vivo half-life or duration of action. A sustained-release form may
result from modification of the active ingredient, such as
modifications that extend circulation residence time, decrease
rates of degradation, decrease rates of clearance or the like, or
may result from formulations or compositions which provide for
extended release of the active ingredient, such as use of various
liposomes, emulsions, micelles, matrices and the like. A
controlled-release form or formulation is a type of
sustained-release form or formulation.
[0063] The terms "synergy", "synergism" or "synergistic" mean more
than the expected additive effect of a combination. A synergistic
effect may be attained when the active ingredients are: (a)
co-formulated and administered or delivered simultaneously in a
combined, unit dosage formulation; (b) delivered as separate
formulations at either then same or different times; or (c) by some
other regimen. A synergistic effect may be attained, by way of
example, by daily administration of one active ingredient, such as
with a half-life of two to six hours, and weekly or longer
administration of another active ingredient, such as with a
half-life of two or greater days.
[0064] A pharmaceutical composition is effective in "treatment of
obesity" or "to induce weight loss" when the composition induces,
over a period of time from 12 to 52 weeks, a statistically
significant and placebo-adjusted decrease in body weight of at
least about 5.0% in a cohort of subjects with a baseline mean BMI
27 kg/m.sup.2.
[0065] The term "therapeutically effective amount" means the amount
of MC4r agonist and GLP-1 receptor agonist, including the ratio of
MC4r agonist and GLP-1 receptor agonist, which will elicit a
biological or medical response in the mammal that is being treated
by a medical doctor or other clinician.
[0066] The terms "treat," "treating" and "treatment," as used
herein, contemplate an action that occurs while a patient is
suffering from, or at risk of suffering from, the specified disease
or disorder, which reduces the onset, incidence or severity of the
disease or disorder.
[0067] Obesity, Diabetes and Related Metabolic Syndrome.
[0068] The compositions and methods disclosed herein can be used
for both medical applications and animal husbandry or veterinary
applications. Typically, the methods are 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 the
present invention involve human patients, but the present invention
may be applied to laboratory, farm, zoo, wildlife, pet, sport or
other animals.
[0069] The combination of MC4r agonist and GLP-1 receptor agonist,
including the ratio of MC4r agonist to GLP-1 receptor agonist and
methods for the administration thereof, is believed to be useful in
prophylaxis and treatment of diseases, disorders and/or conditions
which are energy homeostasis and metabolism related (such as
diabetes, in particular type 2 diabetes; dyslipidemia; fatty liver;
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.
[0070] In one aspect, it is believed that use of a
sustained-release, controlled-release or long acting GLP-1 receptor
agonist in combination with a periodically administered MC4r
agonist is efficacious in treating obesity, diabetes and related
metabolic syndrome. The sustained-release or controlled-release
GLP-1 receptor agonist may be a GLP-1 receptor agonist that has a
comparatively short circulation half-life but is continuously
administered, such as by continuous infusion using a pump device.
Alternatively the sustained-release or controlled-release GLP-1
receptor agonist may have a duration of action greater than about
twenty-four hours, including, but are not limited to, liraglutide,
long acting forms of exenatide, dulaglutide, taspoglutide and
albiglutide. With GLP-1 receptor agonists with a duration of action
greater than about twenty-four hours, the GLP-1 receptor agonists
may be administered by subcutaneous injection on a daily, weekly,
biweekly or other schedule. The MC4r agonist may have a duration of
action less than twelve hours, preferably less than eight hours,
and more preferably less than about six hours, such as a peptide
with a plasma circulation half-life of less than about four hours,
preferably less than about two hours. In this instance, the MC4r
agonist can be administered by subcutaneous injection separately
from the GLP-1 receptor agonist. In one particularly preferred
embodiment, the MC4r agonist is administered prior to periods of
high caloric consumption, such as prior to a meal, such as about
one-half hour to two hours prior to breakfast, lunch, dinner or
evening or other snacking periods. In this embodiment, only one
daily injection of MC4r agonist may be required, or alternatively
two daily injections may be administered.
[0071] The combination of MC4r agonist and GLP-1 receptor agonist,
including the ratio of MC4r agonist and GLP-1 receptor agonist, the
relative durations of action of each of the MC4r agonist and GLP-1
receptor agonist in combination with administration schedules, and
methods for the administration thereof, are particularly believed
to be useful for treatment of body weight related diseases,
disorders and/or conditions characterized by excess body weight,
including obesity and overweight (by promotion of weight loss,
maintenance of weight loss, and/or prevention of weight gain,
including medication-induced weight gain or weight gain subsequent
to cessation of smoking), and diseases, 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.
[0072] 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 meters
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.sub.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) 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 cutoffs 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 cardiovascular 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 methods, combinations and compositions of the
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.
[0073] Methods of Administration and Use.
[0074] The method of administration and use of the combination of
MC4r agonist and GLP-1 receptor agonist, including the ratio of
MC4r agonist and GLP-1 receptor agonist and methods for the
administration thereof, varies depending upon the characteristics
of the drug components utilized in the present invention, the
disease, indication, condition or syndrome to be treated, and other
factors known to those in the art. In general, any method of
administration and use known in the art or hereafter developed may
be employed in the present invention. Without limiting the
foregoing, the following methods of administration and use have
specific application for the indicated indications.
[0075] Compositions comprising the combination of MC4r agonist and
GLP-1 receptor agonist, including the ratio of MC4r agonist and
GLP-1 receptor agonist and methods for the administration thereof,
utilized in the present invention may administered by any suitable
means for therapy, including prophylactic therapy, of obesity,
diabetes and/or metabolic syndrome. In one aspect, the composition
is formulated for subcutaneous injection, and a subcutaneous
injection is given one or more times each day, preferably prior to
a period of high caloric intake, such as a meal, more preferably
between about one and about three hours prior to a period of high
caloric intake, such as a meal. In another aspect, the composition
is formulated as an injectable controlled-release or
sustained-release formulation. In one embodiment, the MC4r agonist
and GLP-1 receptor agonist utilized in the present invention is
formulated with a polyethylene glycol, such as polyethylene 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 the MC4r agonist and
GLP-1 receptor agonist utilized in the present invention are
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. 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 the MC4r agonist and GLP-1 receptor
agonist utilized in the present 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
controlled-release or sustained-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
peptide, the biodegradation rate of the polymer, and other factors
known to those of skill in the art.
[0076] In another and preferred embodiment, different routes of
administration are employed for the administration of each of the
MC4r agonist and GLP-1 receptor agonist. Thus the GLP-1 receptor
agonist and the MC4r agonist may each independently be administered
by any known method, such as subcutaneous administration,
intranasal administration, intravenous administration,
intramuscular administration, oral administration, buccal
administration, sublingual administration, transdermal
administration or other means known in the art, but the method of
administration may be different for the MC4r agonist and the GLP-1
receptor agonist.
[0077] In one aspect, the invention provides a composition
comprising a MC4r agonist and a GLP-1 receptor agonist. The
composition may have a synergistic anti-obesity effect. The
composition may also include a sustained-release formulation. In
the composition, at least one of the MC4r agonist and the GLP-1
receptor agonist may have, upon administration, a duration of
action of at least twenty-four hours. Alternatively, in the
composition both of the MC4r agonist and the GLP-1 receptor agonist
have, upon administration, a duration of action of at least
twenty-four hours.
[0078] In another aspect, the invention provides a method of
treating a patient with obesity, comprising administering to the
patient a combination of (a) a MC4r agonist and (b) a GLP-1
receptor agonist. In the method, at least one of the combination of
(a) a MC4r agonist and (b) a GLP-1 receptor agonist is in a
therapeutically effective amount with respect to treatment of
obesity or inducing weight loss. Alternatively, in the method each
of the (a) MC4r agonist and (b) GLP-1 receptor agonist are
administered in an amount that is not effective with respect to
treatment of obesity or inducing weight loss if administered alone.
The combination may elicit a synergistic effect on treatment of
obesity. Alternatively, if the compounds induce minimal weight loss
the combination may elicit an additive effect on treatment of
obesity. The method may further comprise a sustained-release
formulation. In the method, at least one of the MC4r agonist and
the GLP-1 receptor agonist may have, upon administration, a
duration of action of at least twenty-four hours. Alternatively, in
the method both of the MC4r agonist and the GLP-1 receptor agonist
may have, upon administration, a duration of action of at least
twenty-four hours.
[0079] In another aspect, the invention provides a composition
comprising a combination of a MC4r agonist and a GLP-1 receptor
agonist whereby such MC4r agonist and a GLP-1 receptor agonist are
in such respective proportions as to decrease insulin levels at a
selected time after administration as compared to insulin levels
prior to initial administration of the composition. The decrease in
insulin levels from baseline can be at least about ten percent,
between about ten percent and about fifty percent, at least about
twenty-five percent or at least about fifty percent. The decrease
in insulin levels from baseline in a population administered the
composition is greater than the decrease in insulin levels in a
corresponding population administered either a MC4r agonist or a
GLP-1 receptor agonist but not both, including where the
corresponding population is administered the same quantity of
either a MC4r agonist or a GLP-1 receptor agonist as is
administered to the population receiving the composition. The
composition may comprise a sustained-release formulation. At least
one of the MC4r agonist and the GLP-1 receptor agonist in the
composition may have, upon administration, a duration of action of
at least twenty-four hours. Alternatively, both the MC4r agonist
and the GLP-1 receptor agonist have, upon administration, a
duration of action of at least twenty-four hours.
[0080] In another aspect, the invention provides a pharmaceutical
combination comprising as components (a) a MC4r agonist, and (b) a
GLP-1 receptor agonist; wherein components (a) and (b) are present
in such weight or molar ratio that the composition exerts a
synergistic effect upon administration to a patient. The
pharmaceutical composition may comprise at least one
pharmaceutically acceptable carrier. There is thus provided a
pharmaceutical dosage form comprising a pharmaceutical combination
as described and at least one pharmaceutically acceptable carrier.
The dosage form may be suitable for oral, intravenous,
intraarterial, intraperitoneal, intradermal, transdermal,
intrathecal, intramuscular, intranasal, transmucosal, subcutaneous,
or rectal administration. In the dosage form, at least one of the
components (a) and (b) may be present in sustained-release form, or
alternatively both of the components (a) and (b) may be present in
sustained-release form. The dosage form may further comprise a
sustained-release formulation. In the dosage form, at least one of
the MC4r agonist and the GLP-1 receptor agonist may have, upon
administration, a duration of action of at least twenty-four hours,
or alternatively, both of the MC4r agonist and the GLP-1 receptor
agonist may have, upon administration, a duration of action of at
least twenty-four hours. The pharmaceutical combination may be
utilized in a method of treating obesity, diabetes or metabolic
syndrome in a mammal in need thereof, said method comprising
administering to said mammal a pharmacologically effective amount
of a pharmaceutical combination.
[0081] In another aspect, the invention provides a method of
treating obesity, diabetes or metabolic syndrome in a mammal in
need thereof, comprising separate administration to the patient of
a MC4r agonist and a GLP-1 receptor agonist. In the method, the
quantity and schedule of administration of the MC4r agonist and the
GLP-1 receptor agonist may together be sufficient to produce
synergistic effect, or alternatively additive effect, in the
treatment of obesity, diabetes or metabolic syndrome.
Alternatively, in the method the quantity and schedule of
administration of the MC4r agonist and the GLP-1 receptor agonist
may together be sufficient to initiate a desired pharmacological
response in the treatment of obesity, diabetes or metabolic
syndrome in the mammal. In the method, the desired pharmacological
response may comprise a decrease in body weight, a decrease in body
mass index, a decrease in glucose levels or a decrease in insulin
levels, or a combination of the foregoing. In the practice of the
method, the MC4r agonist and the GLP-1 receptor agonist may be
administered simultaneously to the mammal, may be administered
sequentially to the mammal in either order and may be administered
via different pathways of administration. The MC4r agonist may be
administered by subcutaneous bolus injection and the GLP-1 receptor
agonist may be administered by continuous infusion. Continuous
infusion of the GLP-1 receptor agonist may comprise implantation of
an osmotic delivery device, subcutaneous infusion, or both. The
subcutaneous bolus injection of the MC4r agonist may comprise
subcutaneous injection no more than twice daily, or alternatively
no more than once daily. The MC4r agonist may comprise a
sustained-release form. Similarly, the GLP-1 receptor agonist that
is administered may be a sustained-release 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. In one
embodiment, where a sustained release GLP-1 receptor agonist is
employed, the MC4r agonist is not sustained-release. In such
embodiment, the MC4r agonist may be administered no more than about
twice per day, or alternatively no more than once per day. In such
embodiment, the MC4r agonist which is not sustained-release may
have a plasma circulation half-life of less than about 6 hours, or
alternatively of less than about 3 hours or less than about 2
hours. In another embodiment, the MC4r agonist is administered by
intramuscular injection of a sustained-release form and the GLP-1
receptor agonist is administered by continuous infusion. Continuous
infusion of the GLP-1 receptor agonist may comprise implantation of
an osmotic delivery device.
[0082] In another aspect, there is provided a method of treating
obesity, diabetes or metabolic syndrome in a patient, comprising
bolus parenteral administration of a quantity of a MC4r agonist and
infusion administration of a quantity of GLP-1 receptor agonist,
wherein the quantity of MC4r agonist and the quantity of GLP-1
receptor agonist are together sufficient to initiate a desired
pharmacological response in the treatment of obesity, diabetes or
metabolic syndrome in the patient. In this method, the desired
pharmacological response may comprise a decrease in body weight, a
decrease in body mass index, a decrease in glucose levels or a
decrease in insulin levels, or a combination of the foregoing. The
quantity of MC4r agonist and the quantity of GLP-1 receptor agonist
may together be sufficient to produce synergistic effect in the
treatment of obesity, diabetes or metabolic syndrome in the
patient. In another embodiment, the quantity of each of the MC4r
agonist and the GLP-1 receptor agonist is not sufficient, if either
is administered as monotherapy, to initiate the desired
pharmacological response in the treatment of obesity, diabetes or
metabolic syndrome in the patient. Alternatively, in the method the
quantity of MC4r agonist administered may not be sufficient to
initiate the desired pharmacological response in the treatment of
obesity, diabetes or metabolic syndrome in the patient when
administered as a monotherapy. Alternatively, the quantity of GLP-1
receptor agonist administered may not be sufficient to initiate the
desired pharmacological response in the treatment of obesity,
diabetes or metabolic syndrome in the patient when administered as
a monotherapy. In the method, the bolus parenteral administration
of a quantity of a MC4r agonist may comprise subcutaneous
administration. The quantity of a MC4r agonist may comprise a
sustained-release form, or alternatively does not comprise a
sustained-release form. Subcutaneous administration of the quantity
of a MC4r agonist is no more than two times per day, or
alternatively no more than one time per day. In the event of
subcutaneous administration of a quantity of a MC4r agonist, the
sustained-release form is administered no more than one time per
day, or alternatively between about once per day and once per
fifteen days or once per five days and once per ten days. Infusion
administration of a quantity of GLP-1 receptor agonist may comprise
implantation of an osmotic delivery device. In one embodiment, the
combination of bolus parenteral administration of the quantity of a
MC4r agonist and infusion administration of the quantity of GLP-1
receptor agonist elicits a synergistic effect on treatment of
obesity, diabetes or metabolic syndrome in the patient, or
alternatively elicits an additive effect on treatment of obesity,
diabetes or metabolic syndrome in the patient.
[0083] In another aspect, the invention provides a method of
decreasing side effects associated with therapeutic agents for
treatment of glycemic control, obesity, diabetes or metabolic
syndrome in a patient, comprising administration of a quantity of
MC4r agonist, wherein the quantity of MC4r agonist administered is
not sufficient to initiate the desired pharmacological response in
treating at least one condition from the group comprising glycemic,
obesity, diabetes and metabolic syndrome in the patient when
administered as a monotherapy, and administration of a quantity of
GLP-1 receptor agonist, wherein the quantity of GLP-1 receptor
agonist administered is not sufficient to initiate the desired
pharmacological response in treating at least one condition from
the group comprising glycemic control, obesity, diabetes and
metabolic syndrome in the patient when administered as a
monotherapy, and wherein the quantity of the MC4r agonist 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 glycemic control, obesity,
diabetes and metabolic syndrome in the patient, thereby reducing
side effects in the treatment of at least one of glycemic control,
obesity, diabetes or metabolic syndrome in the patient. By way of
example only, a patient that cannot tolerate doses of a GLP-1
receptor agonist sufficient to achieve the desired glycemic control
may, through the use of a tolerable but ineffective dose of a GLP-1
receptor agonist in combination with an MC4r agonist not
sufficient, if used alone, to achieve the desired glycemic control,
obtain the desired glycemic control.
[0084] In another aspect, the invention provides a method of
treating obesity, diabetes or metabolic syndrome in a patient
either not responsive to or marginally responsive to a GLP-1
receptor agonist, comprising the steps of establishing a dose of a
GLP-1 receptor agonist at which the patient has a marginal rate of
responsiveness with respect to treating obesity, diabetes or
metabolic syndrome with the GLP-1 receptor agonist, administration
of the marginally responsive dose of the GLP-1 receptor agonist,
and administration of a MC4r agonist, wherein the dose of the GLP-1
receptor agonist and the quantity of MC4r agonist are effective to
increase responsiveness with respect to treating obesity, diabetes
or metabolic syndrome. In such method, administration of the
marginally responsive dose of the GLP-1 receptor agonist may
comprise continuous infusion. In such method, administration of the
MC4r agonist may comprise bolus parenteral administration,
including subcutaneous administration. Administration of the MC4r
agonist may alternatively comprise a sustained-release form,
including intramuscular injection of a sustained-release form. In
one embodiment of the method, the combination may elicit a
synergistic effect on treatment of obesity, or alternatively may
elicit an additive effect on treatment of obesity. In another
embodiment of the method, the GLP-1 receptor agonist administered
is a sustained-release GLP-1 receptor agonist, optionally with a
duration of action of at least about twenty-four hours, or
alternatively of at least about one week or at least about two
weeks. In the practice of the method, in one embodiment the MC4r
agonist is not sustained-release, and is administered no more than
about twice per day, or alternatively no more than once per day.
The MC4r agonist which is not sustained-release may have a plasma
circulation half-life of less than about 6 hours, or less than
about 3 hours, or less than about 2 hours.
[0085] Salt Form(s) of MC4r Agonist and GLP-1 Receptor Agonist
Utilized in the Present Invention.
[0086] The MC4r agonist and GLP-1 receptor agonist utilized in the
present invention may be in the form of any pharmaceutically
acceptable salt. The term "pharmaceutically acceptable salts"
refers to 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
aluminum, ammonium, calcium, copper, ferric, ferrous, lithium,
magnesium, manganic salts, 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.
[0087] When the MC4r agonist and GLP-1 receptor agonist utilized in
the present invention are 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, 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 MC4r
agonist and GLP-1 receptor agonist utilized in the present
invention can be prepared in a suitable solvent for the peptide and
an excess of an acid, such as hydrochloric, hydrobromic, sulfuric,
phosphoric, acetic, trifluoroacetic, citric, tartaric, maleic,
succinic or methanesulfonic acid. The acetate and trifluoroacetic
acid salt forms are especially useful. Where MC4r agonist or GLP-1
receptor agonist utilized in the present 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. It
is also to be understood that certain peptides can exist in
solvated forms, including solvates of the free peptide or solvates
of a salt of the compound, as well as unsolvated forms. The term
"solvate" is used herein to describe a molecular complex comprising
one or more compounds utilized in the invention and one or more
pharmaceutically acceptable solvent molecules, for example,
ethanol. The term "hydrate" is employed when said solvent is water.
It is to be understood that all polymorphs, including mixtures of
different polymorphs, are included within the scope of the peptides
utilized in the invention.
[0088] Pharmaceutical Compositions.
[0089] In one aspect, the invention provides a pharmaceutical
composition that includes an MC4r peptide and a GLP-1 receptor
agonist peptide utilized in the present invention and a
pharmaceutically acceptable carrier. The carrier may be a liquid
formulation, and is preferably a buffered, isotonic, aqueous
solution. Pharmaceutically acceptable carriers also include
excipients, such as diluents, carriers and the like, and additives,
such as stabilizing agents, preservatives, solubilizing agents,
buffers and the like, as hereafter described.
[0090] Depending on the desired route of administration, and
whether the MC4r agonist and GLP-1 receptor agonist are formulated
together or formulated separately, the formulation(s) of a
composition including at least one of the MC4r agonist and GLP-1
receptor agonist utilized in the present invention may be varied.
Thus the formulation may be suitable for subcutaneous injection,
intravenous injection, topical applications, ocular applications,
nasal spray applications, inhalation applications, other
transdermal applications and the like.
[0091] The MC4r and GLP-1 peptide compositions utilized in the
present invention may be formulated or compounded into
pharmaceutical compositions that include at least one MC4r agonist
peptide and at least one GLP-1 receptor agonist peptide together
with one or more pharmaceutically acceptable carriers, including
excipients, such as diluents, carriers and the like, and additives,
such as stabilizing agents, preservatives, solubilizing agents,
buffers and the like, as may be desired. Formulation excipients may
include polyvinylpyrrolidone, gelatin, hydroxyl propyl cellulose,
acacia, polyethylene glycol, mannitol, sodium chloride and sodium
citrate. 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, cellulose
derivatives, 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. For pharmaceutical formulations, it
is also contemplated that any of a variety of measured-release,
controlled-release, slow-release or sustained-release formulations
and additives may be employed, so that the dosage may be formulated
so as to provide delivery of an MC4r agonist peptide and GLP-1
receptor agonist peptide utilized in the present invention over a
period of time.
[0092] In general, the actual quantity of MC4r agonist peptide and
GLP-1 receptor agonist peptide utilized in the present invention
administered to a patient will vary between fairly wide ranges
depending on the mode of administration, the formulation used, and
the response desired.
[0093] In practical use, the peptides of the invention as the
active ingredient in an admixture with a pharmaceutical carrier
according to conventional pharmaceutical compounding techniques.
The carrier may take a wide variety of forms depending on the form
of preparation desired for administration, for example, oral,
parenteral (including intravenous), urethral, vaginal, nasal,
buccal, sublingual, or the like.
[0094] MC4r agonist peptides and GLP-1 receptor agonist peptides
may be administered parenterally. Solutions or suspensions of these
active peptides 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.
[0095] The pharmaceutical forms suitable for injectable use include
sterile aqueous solutions or dispersions and sterile powders 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.
[0096] The MC4r agonist peptides and GLP-1 receptor agonist
peptides utilized in the present invention may be therapeutically
applied by means of nasal administration. By "nasal administration"
is meant any form of intranasal administration of any of the
peptides utilized in the present invention. The peptides may be in
an aqueous solution, such as a solution including saline, citrate
or other common excipients or preservatives. The peptides may also
be in a dry or powder formulation.
[0097] The MC4r agonist peptides and GLP-1 receptor agonist
peptides utilized in the present invention may be formulated with
any of a variety of agents that increase effective nasal absorption
of drugs, including peptide drugs. These agents may increase nasal
absorption without unacceptable damage to the mucosal membrane.
U.S. Pat. Nos. 5,693,608, 5,977,070 and 5,908,825, among others,
teach a number of pharmaceutical compositions that may be employed,
including absorption enhancers, and the teachings of each of the
foregoing, and all references and patents cited therein, are
incorporated by reference.
[0098] If in an aqueous solution, the peptides comprising the
formulation may be appropriately buffered by means of saline,
acetate, phosphate, citrate, acetate or other buffering agents,
which may be at any physiologically acceptable pH, generally from
about pH 4 to about pH 7. A combination of buffering agents may
also be employed, such as phosphate buffered saline, a saline and
acetate buffer, and the like. In the case of saline, a 0.9% saline
solution may be employed. In the case of acetate, phosphate,
citrate, and the like, a 50 mM solution may be employed. In
addition to buffering agents, a suitable preservative may be
employed, to prevent or limit bacteria and other microbial growth.
One such preservative that may be employed is 0.05% benzalkonium
chloride.
[0099] The MC4r peptides utilized in the present invention may be
therapeutically administered by means of an injection of a
controlled-release or sustained-release formulation. In one
embodiment, an MC4r agonist peptide utilized in the present
invention is formulated for a deep intramuscular injection, such as
in the gluteal or deltoid muscle, of a formulation with a
polyethylene glycol, such as polyethylene 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 an MC4r agonist peptide utilized in the
present 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. In one embodiment poly
(D,L-lactide-co-glycolide) polymer is employed. In general, any of
a number of injectable and bioerodible polymers, which are
preferably also adhesive polymers, may be employed in a
controlled-release or sustained-release injectable formulation.
Alternatively other controlled-release or sustained-release
formulations may be employed, including formulations permitting
subcutaneous injection, which other formulations may include one or
more of nano/microspheres (such as compositions including PLGA
polymers), liposomes, emulsions (such as water-in-oil emulsions),
gels, insoluble salts or suspensions in oil. The formulation may be
such that an injection is required on a daily, weekly, monthly or
other periodic basis, depending on the concentration and amount of
cyclic peptide, the controlled- or sustained-release rate of the
materials employed, and other factors known to those of skill in
the art.
[0100] Particularly preferred are GLP-1 receptor agonist peptides
which are long-acting, such as those GLP-1 receptor agonist
peptides with a duration of action greater than about twenty-four
hours. Representative examples of such GLP-1 receptor agonist
peptides with a duration of action greater than about twenty-four
hours include, but are not limited to, liraglutide, long acting
forms of exenatide, dulaglutide, taspoglutide and albiglutide.
[0101] The GLP-1 receptor agonist peptides utilized in the present
invention may be therapeutically administered by means of an
injection of a controlled-release or sustained-release formulation.
In one embodiment, a GLP-1 receptor agonist utilized in the present
invention is formulated for a deep intramuscular injection, such as
in the gluteal or deltoid muscle, of a formulation with a
polyethylene glycol, such as polyethylene 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 GLP-1 receptor agonist utilized in
the present 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. In one embodiment poly
(D,L-lactide-co-glycolide) polymer is employed. In general, any of
a number of injectable and bioerodible polymers, which are
preferably also adhesive polymers, may be employed in a
controlled-release or sustained-release injectable formulation.
Alternatively other controlled-release or sustained-release
formulations may be employed, including formulations permitting
subcutaneous injection, which other formulations may include one or
more of nano/microspheres (such as compositions including PLGA
polymers), liposomes, emulsions (such as water-in-oil emulsions),
gels, insoluble salts or suspensions in oil. The formulation may be
such that an injection is required on a daily, weekly, monthly or
other periodic basis, depending on the concentration and amount of
cyclic peptide, the controlled- or sustained-release rate of the
materials employed, and other factors known to those of skill in
the art.
[0102] Routes of Administration.
[0103] If a composition including one or more MC4r agonist peptides
utilized in the present invention is administered by injection, the
injection may be intravenous, subcutaneous, intramuscular,
intraperitoneal or other means known in the art. The MC4r agonist
peptides utilized in the present invention may be formulated by any
means known in the art, including but not limited to formulation as
tablets, capsules, caplets, suspensions, powders, lyophilized
preparations, suppositories, ocular drops, skin patches, oral
soluble formulations, sprays, aerosols and the like, and may be
mixed and formulated with buffers, binders, excipients,
stabilizers, anti-oxidants and other agents known in the art. In
general, any route of administration by which the peptides of
invention are introduced across an epidermal layer of cells may be
employed. Administration means may thus include administration
through mucous membranes, buccal administration, oral
administration, dermal administration, inhalation administration,
nasal administration, urethral administration, vaginal
administration, and the like.
[0104] In one aspect the GLP-1 receptor agonist is a long acting
form of GLP-1 receptor agonist, such as form of GLP-1 with an
extended half-life or reduced clearance rate, or a
controlled-release or sustained-release formulation. In a preferred
embodiment, the GLP-1 receptor agonist has a duration of action
greater than about twenty-four hours, including liraglutide and
long acting forms of exenatide, dulaglutide, taspoglutide and
albiglutide. Particularly preferred are GLP-1 receptor agonists
which are not required to be administered more frequently than
about once per day, once per week, once per every two weeks or one
per month. Alternatively, the GLP-1 receptor agonist may have a
short half-life, but may be continuously or intermittently
administered, such as by means of an infusion device or depot
formulation that allows slow release and gradual absorption.
[0105] For sustained-released forms of formulations of GLP-1
receptor agonist, in one embodiment a continuous delivery method
and/or device may be employed. Such methods or devices may include
intravenous infusion, which may be by gravity (such as using a
collapsible plastic bag) or by an infusion pump. A variety of
implantable devices and/or methods for continuous delivery are
known, including but not limited to use of implantable devices
(which may deliver active ingredient intravenously or
subcutaneously). One type of implantable device that may be
employed is an osmotic delivery device, such as that disclosed in
U.S. Pat. No. 8,298,561.
[0106] Regardless of the formulation or route of administration
employed for administration of GLP-1 receptor agonist, the MC4r
agonist may be administered separately, and in a preferred
embodiment is administered separately. Such separate administration
may be, for example, by subcutaneous injection or transdermal
administration. Intermittent delivery devices, including
implantable intermittent delivery devices, may also be employed,
which deliver, for example, a bolus quantity of MC4r agonist on a
determined or variable schedule, such as once daily, twice daily or
in conjunction with one or more periods of high caloric intake.
[0107] Therapeutically Effective Amount.
[0108] In general, the actual quantity of MC4r agonist and GLP-1
receptor agonist utilized in the present invention administered to
a patient will vary between fairly wide ranges depending upon the
mode of administration, characteristics of each of the MC4r agonist
and GLP-1 receptor agonist, including pharmacokinetics and
pharmacodynamics, 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 MC4r agonist
and an amount, which may be separately administered of GLP-1
receptor agonist. Preferably the therapeutically effective amount
of MC4r agonist and GLP-1 receptor agonist results in measures of
one or more parameters, such as insulin levels, blood glucose
levels or other metabolic parameters, that are clinically superior
to the measures of the same parameters obtained with MC4r agonist
administered without administration of GLP-1 receptor agonist, or
alternatively measures of the same parameters obtained with GLP-1
receptor agonist administered without the administration of MC4r
agonist. Further preferably the therapeutic effective amount of
MC4r agonist and GLP-1 receptor agonist results in a synergistic
effect in the treatment of obesity, diabetes or metabolic syndrome,
and specifically in at least one of inducing weight loss or
inducing glycemic control.
[0109] In one aspect, a therapeutically effective amount is
determined, in part, by selection of a dose of MC4r agonist and
GLP-1 receptor agonist such that side effects are limited with
respect to each of the MC4r agonist and GLP-1 receptor agonist. For
example, a patient may be able to achieve desired glycemic control
with a dose of GLP-1 receptor agonist which nonetheless induces
undesired side effects, such as gastrointestinal symptoms or
adverse reactions such as nausea, diarrhea, vomiting, constipation
or the like. A lower dose of GLP-1 receptor agonist may not achieve
desired glycemic control in the patient. However, the combination
of the lower dose of GLP-1 receptor agonist together with a dose of
MC4r agonist, which dose of MC4r agonist may not be sufficient to
induce weight loss, are together sufficient to achieve the desired
glycemic control without inducing undesired side effects, or
inducing only reduced or tolerable side effects.
[0110] GLP-1 Receptor Agonists Utilized in the Present
Invention.
[0111] 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.
[0112] MC4r Peptides Utilized in the Present Invention.
[0113] Preferred MC4r agonist peptides with utility in the current
invention include the peptides disclosed in U.S. patent application
Ser. No. 13/311,824, entitled "Melanocortin Receptor-Specific
Peptides", filed Jul. 11, 2014, and issued as U.S. Pat. No.
8,846,601 on Sep. 30, 2014; U.S. patent application Ser. No.
14/328,995, entitled "Melanocortin Receptor-Specific
Heptapeptides", filed Dec. 6, 2011, and issued as U.S. Pat. No.
8,846,601 on Sep. 30, 2014; International Application No.
PCT/US2010/037589, published as International Publication No. WO
2010/144344, entitled "Melanocortin Receptor-Specific Peptides",
filed on Jun. 7, 2010; and U.S. patent application Ser. No.
12/952,238, entitled "Melanocortin Receptor-Specific Peptides for
Treatment of Sexual Dysfunction", filed on Nov. 23, 2010 and issued
as U.S. Pat. No. 8,487,073 on Jul. 16, 2013.
[0114] In one aspect, the invention provides a cyclic heptapeptide
which contains a core sequence derived from His-Phe-Arg-Trp (SEQ ID
NO:3) within the cyclic portion, and where the amino acid in the
first position is outside the cyclic portion and has a side chain
including at least one primary amine, guanidine or urea group.
Representative amino acids which may be in the first position
include, but are not limited to, Dap, Dab, Orn, Lys, Cit or
Arg.
[0115] The core sequence derived from His-Phe-Arg-Trp will include
unsubstituted D-Phe, D-Nal 1 or D-Nal 2 in the Phe position, but
typically a variety of amino acids may be utilized for the
remaining amino acids in the core sequence. In general, the His
position may be a substituted or unsubstituted Pro, or may be an
amino acid with a side chain including at least one primary amine,
secondary amine, alkyl, cycloalkyl, cycloheteroalkyl, aryl,
heteroaryl, alcohol, ether, sulfide, sulfone, sufoxide, carbomyl or
carboxyl. The Arg position may be a substituted or unsubstituted
Pro, or may be an amino acid with a side chain including at least
one primary amine, secondary amine, guanidine, urea, alkyl,
cycloalkyl, cycloheteroalkyl, aryl, heteroaryl, or ether. The Trp
position may be an amino acid with a side chain including at least
one substituted or unsubstituted aryl or heteroaryl.
[0116] Lactam-bridges are preferred for making the peptide cyclic,
but other bridging groups are possible and contemplated, including
specifically the groups:
--(CH.sub.2).sub.x--C(.dbd.O)--NH--(CH.sub.2).sub.y--,
--(CH.sub.2).sub.x--NH--C(.dbd.O)--(CH.sub.2).sub.y--,
--(CH.sub.2).sub.x--S--S--(CH.sub.2).sub.y--,
--(CH.sub.2).sub.x--C(.dbd.O)--(CH.sub.2).sub.z--C(.dbd.O)--(CH.sub.2).s-
ub.y--,
--(CH.sub.2).sub.x--C(.dbd.O)--NH--C(.dbd.O)--(CH.sub.2).sub.y--,
or
--(CH.sub.2).sub.x--NH--C(.dbd.O)--NH--(CH.sub.2).sub.y--;
where x and y are each independently 1 to 5. For certain
indications and uses, including without limitation for peptides
more selective for MC4r than for MC1r or, in general, for other
melanocortin receptors, particularly preferred are the bridging
groups --(CH.sub.2).sub.2--C(.dbd.O)--NH--(CH.sub.2).sub.3-- and
--(CH.sub.2).sub.2--NH--C(.dbd.O)--(CH.sub.2).sub.2--. In this
context, the teachings of U.S. patent application Ser. No.
13/311,817, filed on Dec. 6, 2011, entitled "Lactam-Bridged
Melanocortin Receptor-Specific Peptides", and International
Application No. PCT/US2010/037584, filed on Jun. 7, 2010 and
published under the same title as International Publication No. WO
2010/144341, are incorporated herein by reference as if set forth
in full.
[0117] The MC4r agonist peptides may contain one or more asymmetric
elements such as stereogenic centers, stereogenic axes and the
like, so that the peptides can exist in different stereoisomeric
forms. For both specific and generically described peptides all
forms of isomers at all chiral or other isomeric centers, including
enantiomers and diastereomers, are intended to be covered herein.
The peptides may each include multiple chiral centers, and may be
used as a diastereomeric mixture or an enantiomerically enriched
mixture, in addition to use of the peptides of the invention in
enantiopure preparations. Typically, the peptides will be
synthesized with the use of chirally pure reagents, such as
specified L- or D-amino acids, using reagents, conditions and
methods such that enantiomeric purity is maintained, but it is
possible and contemplated that diastereomeric mixtures may be made.
Such racemic mixtures may optionally be separated using well-known
techniques and an individual enantiomer may be used alone. In cases
and under specific conditions of temperature, solvents and pH
wherein peptides may exist in tautomeric forms, each tautomeric
form is contemplated as being included within this invention
whether existing in equilibrium or predominantly in one form. Thus
a single enantiomer of a peptide, which is an optically active
form, can be obtained by asymmetric synthesis, synthesis from
optically pure precursors, or by resolution of the racemates.
[0118] The invention is further intended to include prodrugs of the
present peptides, which on administration undergo chemical
conversion by metabolic processes before becoming active
pharmacological peptides. In general, such prodrugs will be
functional derivatives of the present peptides, which are readily
convertible in vivo into an MC4r peptide. Prodrugs are any
covalently bonded compounds, which release the active parent
peptide drug 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.
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. Thus by
way of example and not limitation, a prodrug includes peptides
wherein an ester prodrug form is employed, such as, for example,
lower alkyl esters of an R group of the peptide, such as where R is
--OH, which lower alkyl esters may include from 1-8 carbons in an
alkyl radical or aralkyl esters which have 6-12 carbons in an
aralkyl radical. 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 peptide drug in vivo.
[0119] The subject invention also includes peptides in which one or
more atoms 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 and oxygen, such as .sup.2H, .sup.3H, .sup.13C, .sup.14C,
.sup.15N, .sup.18O and .sup.17O, respectively. MC4r agonist
peptides utilized in the present invention and pharmaceutically
acceptable salts or solvates which contain the aforementioned
isotopes and/or other isotopes of other atoms are within the scope
of this invention. Certain isotopically-labeled compounds, 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 in 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 peptides can generally be
prepared by substituting an isotopically labeled reagent for a
non-isotopically labeled reagent.
[0120] Tests and Assays Employed in Evaluation of MC4r Peptides of
the Present Invention.
[0121] The MC4r agonist peptides may be tested by a variety of
assay systems and animal models to determine binding, functional
status and efficacy.
[0122] A competitive inhibition binding assay may be performed
using membrane homogenates prepared from HEK-293 cells that express
recombinant hMC4r, hMC3r, or hMC5r, and from B-16 mouse melanoma
cells (containing endogenous MC1r). In some instances, HEK-293
cells that express recombinant hMC1r are employed. In the examples
that follow, all MC3r, MC4r and MC5r values are for human
recombinant receptors. MC1r values are for B-16 mouse melanoma
cells, unless the heading is "hMC1r", in which case the value is
for human recombinant MC1r. Assays were performed in 96 well GF/B
Millipore multiscreen filtration plates (MAFB NOB10) pre-coated
with 0.5% bovine serum albumin (Fraction V). Membrane homogenates
were incubated with 0.2 nM (for hMC4r) 0.4 nM (for MC3r and MC5r)
or 0.1 nM (for mouse B16 MC1r or hMC1r) [I.sup.125]-NDP-.alpha.-MSH
(Perkin Elmer) and increasing concentrations of test peptides of
the present invention 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 minutes at 37.degree. C., the assay mixture was
filtered and the membranes washed three times with ice-cold buffer.
Filters were dried and counted in a gamma counter for bound
radioactivity. Non-specific binding was 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%) was 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 was normalized
with respect to 100% specific binding to determine the percent
inhibition of [I.sup.125]-NDP-.alpha.-MSH binding. Each assay was
conducted in triplicate and the actual mean values are described,
with results less than 0% reported as 0%. Ki values for test
peptides of the present invention were determined using Graph-Pad
Prism.RTM. curve-fitting software.
[0123] Alternatively, a competitive inhibition binding assay may be
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 AcroWell 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.
[0124] Competitive binding studies using [I.sup.125]-AgRP (83-132)
can be conducted using membrane homogenates isolated from cells
that express hMC4r. The assays were performed in 96-well GF/B
Millipore multiscreen filtration plates (MAFB NOB10) pre-coated
with 0.5% bovine serum albumin (Fraction V). The assay mixture
contained 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, 0.5%
bovine serum albumin, membrane homogenates, radioligand
[I.sup.125]-AgRP (83-132) (Perkin Elmer) and increasing
concentrations of peptides of the present invention in a total
volume of 200 .mu.L. Binding was measured at radioligand
concentrations of 0.2 nM. After incubating for 1 hour at 37.degree.
C., the reaction mixture was filtered and washed with assay buffer
containing 500 mM NaCl. The dried discs were punched out from the
plate and counted on a gamma counter. Ki values for test peptides
of the present invention were determined using Graph-Pad Prism.RTM.
curve-fitting software.
[0125] Accumulation of intracellular cAMP was examined as a measure
of the ability of the MC4r peptides to elicit a functional response
in HEK-293 cells that express MC4r. Confluent HEK-293 cells that
express recombinant hMC4r were detached from culture plates by
incubation in enzyme-free cell dissociation buffer. Dispersed cells
were suspended in Earle's 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.
[0126] Food Intake and Body Weight Change.
[0127] Change in food intake and body weight can be evaluated using
animal models. Male Sprague-Dawley rats are obtained from Hilltop
Lab Animals, Inc. (Scottsdale, Pa.) or other vendors. Animals are
individually housed in conventional polystyrene hanging cages and
maintained on a controlled 12 hour on/off light cycle. Water and
pelleted food is provided ad libitum. The rats are dosed IV with
vehicle or selected peptides (0.3 to 1.0 mg/kg), or dosed
subcutaneously with vehicle or selected peptides (doses up to 30
mg/kg). The changes in body weight and food intake for the 24 hour
period after dosing is determined. The changes in body weight and
food intake for the 48 hour and 72 hour periods after dosing can
also be measured to determine reversal of changes in body weight
and food intake effects back to baseline levels.
[0128] Combination With Third Agent for Certain Indications.
[0129] The combination of MC4r agonist and GLP-1 receptor agonist,
including the ratio of MC4r agonist and GLP-1 receptor agonist
and/or pharmacokinetic properties, including the half-life of MC4r
agonist and GLP-1 receptor agonist, and methods for the
administration thereof, compositions and methods of the present
invention may be used for treatment of any of the foregoing
diseases, indications, conditions or syndromes in combination with
a third additional other pharmaceutically active compound. Such
combination administration may be by means of a single dosage form
which includes both the combination of MC4r agonist and GLP-1
receptor agonist, including the ratio of MC4r agonist and GLP-1
receptor agonist and methods for the administration thereof, of the
present invention and one more other pharmaceutically active
compounds, such single dosage form including a tablet, capsule,
spray, inhalation powder, injectable liquid or the like.
Alternatively, combination administration may be by means of
administration of two different dosage forms, with one dosage form
containing a peptide of the present invention, and the other dosage
form including another pharmaceutically active compound. In this
instance, the dosage forms may be the same or different. Without
meaning to limit combination therapies, the following exemplifies
certain combination therapies which may be employed.
[0130] In particular 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 may
be employed. Drugs that reduce energy intake include, in part,
various pharmacological agents, referred to as anorectic drugs,
which are used as adjuncts to behavioral therapy in weight
reduction programs.
[0131] Generally, a total dosage of the below-described obesity
control agents or medications, when used in combination with one or
more peptides of the present invention can range from 0.1 to 3,000
mg/day, preferably from about 1 to 1,000 mg/day and more preferably
from about 1 to 200 mg/day in single or 2-4 divided doses. The
exact dose, however, is determined by the attending clinician and
is dependent on such factors as the potency of the compound
administered, the age, weight, condition and response of the
patient.
[0132] The combination of MC4r agonist and GLP-1 receptor agonist,
including the ratio of MC4r agonist and GLP-1 receptor agonist and
methods for the administration thereof, of the invention may in
addition or alternatively further be combined with one or more
other pharmacologically active agent(s) that is (are) useful in the
treatment of diseases, 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.
[0133] According to a further aspect of the invention there is
provided a combination treatment comprising the administration of a
pharmacologically effective amount of the combination of MC4r
agonist and GLP-1 receptor agonist, including the ratio of MC4r
agonist and GLP-1 receptor agonist and methods for the
administration thereof, according to the invention, optionally
together with one or more pharmaceutically acceptable diluents or
carriers, with the simultaneous, sequential or separate
administration one or more of the following agents selected from:
[0134] insulin and insulin analogues; [0135] insulin secretagogues,
including sulphonylureas (e.g. glipizide) and prandial glucose
regulators (sometimes called "short-acting secretagogues"), such as
meglitinides (e.g. repaglinide and nateglinide); [0136] agents that
improve incretin action, for example dipeptidyl peptidase IV
(DPP-4) inhibitors (e.g. vildagliptin, saxagliptin, and
sitagliptin); [0137] insulin sensitising agents including
peroxisome proliferator activated receptor gamma (PPAR.gamma.)
agonists, such as thiazolidinediones (e.g. pioglitazone and
rosiglitazone), and agents with any combination of PPAR alpha,
gamma and delta activity; [0138] 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;
[0139] agents designed to reduce/slow the absorption of glucose
from the intestine, such as alpha-glucosidase inhibitors (e.g.
miglitol and acarbose); [0140] agents which antagonise the actions
of or reduce secretion of glucagon, such as amylin analogues (e.g.
pramlintide); [0141] agents that prevent the reabsorption of
glucose by the kidney, such as sodium-dependent glucose transporter
2 (SGLT-2) inhibitors (e.g. dapagliflozin); [0142] 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;
[0143] anti-dyslipidemia agents, such as HMG-CoA reductase
inhibitors (statins, e.g. rosuvastatin) and other
cholesterol-lowering agents; PPARa 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; [0144]
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 dihydropridines
(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); [0145] 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); 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);
[0146] 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); [0147]
feeding behavior modifying agents, such as orexin receptor
modulators and melanin-concentrating hormone (MCH) modulators;
[0148] neuropeptide Y (NPY)/NPY receptor modulators; [0149]
pyruvate dehydrogenase kinase (PDK) modulators; [0150] serotonin
receptor modulators; [0151] leptin/leptin receptor modulators;
[0152] ghrelin/ghrelin receptor modulators; or [0153] monoamine
transmission-modulating agents, such as selective serotonin
reuptake inhibitors (SSRI) (e.g. fluoxetine), noradrenaline
reuptake inhibitors (NARI), noradrenaline-serotonin reuptake
inhibitors (SNRI), triple monoamine reuptake blockers (e.g.
tesofensine), and monoamine oxidase inhibitors (MAOI) (e.g.
toloxatone and amiflamine), 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.
EXAMPLES
[0154] The invention is further exemplified by the following
non-limiting examples:
[0155] 1. The following MC4r agonist peptides were synthesized and
averaged MC4r Ki values for peptides were determined as indicated.
All Ki values were determined using [I.sup.125]-NDP-.alpha.-MSH
unless marked with an "k", in which event the values were
determined using Eu-NDP-.alpha.-MSH. Ki values marked "ND" were not
determined.
TABLE-US-00002 No. Amino Acid Sequence MC4r Ki(nM) 1
Ac-Arg-cyclo(Asp-His-D-Phe-Arg-Trp-Lys)-NH.sub.2 5 2
Ac-Arg-cyclo(Asp-Lys-D-Phe-Arg-Trp-Lys)-NH.sub.2 30 3
Ac-Arg-cyclo(Asp-Ser-D-Phe-Arg-Trp-Lys)-NH.sub.2 172 4
Ac-Arg-cyclo(Asp-Ser-D-Phe-Lys-Trp-Lys)-NH.sub.2 4293 5
Ac-Arg-cyclo(Asp-Ala-D-Phe-Cit-Trp-Lys)-NH.sub.2 2048* 6
Ac-Arg-cyclo(Asp-His-D-Phe-Lys-Trp-Lys)-NH.sub.2 533* 7
Ac-Arg-cyclo(Asp-Lys-D-Phe-Lys-Trp-Lys)-NH.sub.2 2957* 8
Ac-Arg-cyclo(Asp-Ala-D-Phe-Lys-Trp-Lys)-NH.sub.2 3018* 9
Ac-Arg-cyclo(Asp-Ala-D-Phe-Arg-Trp-Lys)-NH.sub.2 19* 10
Ac-Arg-cyclo(Asp-Phe-D-Phe-Arg-Trp-Lys)-NH.sub.2 51* 11
Ac-Arg-cyclo(Asp-Tyr-D-Phe-Arg-Trp-Lys)-NH.sub.2 43* 12
Ac-Arg-cyclo(Asp-Leu-D-Phe-Arg-Trp-Lys)-NH.sub.2 33* 13
Ac-Arg-cyclo(Asp-Nle-D-Phe-Arg-Trp-Lys)-NH.sub.2 30* 14
Ac-Arg-cyclo(Asp-Thr(Bzl)-D-Phe-Arg-Trp-Lys)-NH.sub.2 1 15
Ac-Arg-cyclo(Asp-Hyp(Bzl)-D-Phe-Arg-Trp-Lys)-NH.sub.2 0.7* 16
Ac-Arg-cyclo(Asp-Val-D-Phe-Arg-Trp-Lys)-NH.sub.2 23 17
Ac-Arg-cyclo(Asp-Aic-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 18
Ac-Arg-cyclo(Asp-Pro(4R-Bzl)-D-Phe-Arg-Trp-Lys)-NH.sub.2 3* 19
Ac-Arg-cyclo(Asp-His-D-Nal 1-Arg-Trp-Lys)-NH.sub.2 3 20
Ac-Arg-cyclo(Asp-D-Nle-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 21
Ac-Arg-cyclo(Asp-D-Ala-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 22
Ac-Arg-cyclo(Asp-D-Ser-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 23
Ac-Arg-cyclo(Asp-His-D-Phe-Ala-Trp-Lys)-NH.sub.2 ND 24
Ac-Arg-cyclo(Asp-His-D-Phe-Nle-Trp-Lys)-NH.sub.2 ND 25
Ac-Arg-cyclo(Asp-His-D-Phe-Val-Trp-Lys)-NH.sub.2 ND 26
Ac-Arg-cyclo(Asp-His-D-Phe-Ser-Trp-Lys)-NH.sub.2 ND 27
Ac-Arg-cyclo(Asp-Aib-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 28
Ac-Arg-cyclo(Asp-Arg-D-Phe-Arg-Trp-Lys)-NH.sub.2 4* 29
Ac-Arg-cyclo(Asp-Asn-D-Phe-Arg-Trp-Lys)-NH.sub.2 32* 30
Ac-Arg-cyclo(Asp-Asp-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 31
Ac-Arg-cyclo(Asp-Glu-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 32
Ac-Arg-cyclo(Asp-Gln-D-Phe-Arg-Trp-Lys)-NH.sub.2 13 33
Ac-Arg-cyclo(Asp-Gly-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 34
Ac-Arg-cyclo(Asp-Ile-D-Phe-Arg-Trp-Lys)-NH.sub.2 19* 35
Ac-Arg-cyclo(Asp-Thr-D-Phe-Arg-Trp-Lys)-NH.sub.2 140* 36
Ac-Arg-cyclo(Asp-Trp-D-Phe-Arg-Trp-Lys)-NH.sub.2 37* 37
Ac-Arg-cyclo(Asp-D-Val-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 38
Ac-Arg-cyclo(Asp-Met-D-Phe-Arg-Trp-Lys)-NH.sub.2 35* 39
Ac-Arg-cyclo(Asp-D-Arg-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 40
Ac-Arg-cyclo(Asp-D-Asp-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 41
Ac-Arg-cyclo(Asp-D-His-D-Phe-Arg-Trp-Lys)-NH.sub.2 95* 42
Ac-Arg-cyclo(Asp-D-Leu-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 43
Ac-Arg-cyclo(Asp-D-Lys-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 44
Ac-Arg-cyclo(Asp-D-Thr-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 45
Ac-Arg-cyclo(Asp-D-Trp-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 46
Ac-Arg-cyclo(Asp-D-Thr(Bzl)-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 47
Ac-Arg-cyclo(Asp-D-Cha-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 48
Ac-Arg-cyclo(Asp-Phe(2-C(.dbd.O)--NH.sub.2)-D-Phe-Arg-Trp-Lys)-NH.sub.-
2 23 49
Ac-Arg-cyclo(Asp-Phe-(3-(C(.dbd.O)--NH.sub.2))-D-Phe-Arg-Trp-Lys)-NH.s-
ub.2 15 50
Ac-Arg-cyclo(Asp-Phe(4-(C(.dbd.O)--NH.sub.2))-D-Phe-Arg-Trp-Lys)-NH.su-
b.2 30 51 Ac-Arg-cyclo(Asp-His-D-Phe-Arg-Trp-Lys)-OH 22* 52
Ac-Arg-cyclo(Asp-Ala-D-Phe-Arg-Trp-Lys)-OH 144* 53
Ac-Arg-cyclo(Asp-His-D-Phe-Cit-Trp-Lys)-OH 214* 54
Ac-Arg-cyclo(Asp-Hyp(Bzl)-D-Phe-Arg-Trp-Lys)-OH 3 55
Ac-Arg-cyclo(Asp-Pro(Bn)-D-Phe-Arg-Trp-Lys)-OH 12 56
Ac-Arg-cyclo(Asp-Asn-D-Phe-Arg-Trp-Lys)-OH 100 57
Ac-Arg-cyclo(Asp-Gln-D-Phe-Arg-Trp-Lys)-OH 115 58
Ac-Arg-cyclo(Asp-Orn-D-Phe-Arg-Trp-Lys)-OH 23 59
Ac-Arg-cyclo(Asp-Dap-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 60
Ac-Arg-cyclo(Asp-His-D-(alpha-Me)Phe-Arg-Trp-Lys)-NH.sub.2 47* 61
Ac-Arg-cyclo(Asp-His-(alpha-Me)-Phe-Arg-Trp-Lys)-NH.sub.2 ND 62
Ac-Arg-cyclo(Asp-Dap-D-Phe-Arg-Trp-Lys)-NH.sub.2 80* 63
Ac-Arg-cyclo(Asp-Sar-D-Phe-Arg-Trp-Lys)-NH.sub.2 15 64
Ac-Arg-cyclo(Glu-His-D-Phe-Arg-Trp-Orn)-NH.sub.2 1* 65
Ac-Arg-cyclo(Asp-His-D-Phe-Met(O2)-Trp-Lys)-NH.sub.2 ND 66
Ac-Arg-cyclo(Asp-His-D-Phe-Gln-Trp-Lys)-NH.sub.2 ND 67
Ac-Arg-cyclo(Asp-Orn-D-Phe-Arg-Trp-Lys)-NH.sub.2 7* 68
Ac-Arg-cyclo(Asp-His-D-Phe-D-Nle-Trp-Lys)-NH.sub.2 ND 69
Ac-D-Arg-cyclo(Asp-His-D-Phe-Arg-Trp-Lys)-NH.sub.2 8* 70
Ac-D-Arg-cyclo(Asp-Ala-D-Phe-Arg-Trp-Lys)-NH.sub.2 46* 71
Ac-D-Arg-cyclo(Asp-Gln-D-Phe-Arg-Trp-Lys)-NH.sub.2 45 72
Ac-D-Arg-cyclo(Asp-Pro(4R-Bzl)-D-Phe-Arg-Trp-Lys)-NH.sub.2 2* 73
Ac-Arg-cyclo(Asp-Arg-D-Phe-Arg-Trp-Lys)-OH 60* 74
Ac-Arg-cyclo(Glu-Gln-D-Phe-Arg-Trp-Orn)-NH.sub.2 3 75
Ac-Arg-cyclo(Orn-His-D-Phe-Arg-Trp-Glu)-NH.sub.2 0.6 76
Ac-Arg-cyclo(Asp-Arg-D-Phe-Arg-Trp-Lys)-NH.sub.2 14 77
Ac-Arg-cyclo(Asp-Met-(O.sub.2)-D-Phe-Arg-Trp-Lys)-NH.sub.2 7 78
Ac-Arg-cyclo(Asp-Gln-D-Phe-Arg-Trp-Lys)-NH.sub.2 86* 79
Ac-Arg-cyclo(Asp-Hyp-D-Phe-Arg-Trp-Lys)-NH.sub.2 4 80
Ac-Arg-cyclo(Asp-Pro(4R-NH.sub.2)-D-Phe-Arg-Trp-Lys)-NH.sub.2 5 81
Ac-Arg-cyclo(Glu-Gln-D-Phe-Arg-Trp-Orn)-OH 43 82
Ac-Arg-cyclo(Glu-His-D-Phe-Arg-Trp-Orn)-OH 4 83
Ac-Arg-cyclo(Orn-Gln-D-Phe-Arg-Trp-Glu)-OH ND 84
Ac-Arg-cyclo(Orn-His-D-Phe-Arg-Trp-Glu)-OH 12* 85
Ac-D-Arg-cyclo(Glu-His-D-Phe-Arg-Trp-Orn)-OH 40* 86
Ac-D-Arg-cyclo(Glu-Gln-D-Phe-Arg-Trp-Orn)-OH 66* 87
Ac-D-Arg-cyclo(Glu-Dab-D-Phe-Arg-Trp-Orn)-OH 171* 88
Ac-Arg-cyclo(Glu-Arg-D-Phe-Arg-Trp-Orn)-OH 37* 89
Ac-Arg-cyclo(Glu-Lys-D-Phe-Arg-Trp-Orn)-OH 113* 90
Ac-Arg-cyclo(Glu-Orn-D-Phe-Arg-Trp-Orn)-OH 9 91
Ac-Arg-cyclo(Glu-Asn-D-Phe-Arg-Trp-Orn)-OH 167 92
Ac-Arg-cyclo(Glu-Cit-D-Phe-Arg-Trp-Orn)-OH 184* 93
Ac-Arg-cyclo(Asp-Dab(AcetyI)-D-Phe-Arg-Trp-Lys)-NH.sub.2 33 94
Ac-Arg-cyclo(Asp-Dab(Glycly)-D-Phe-Arg-Trp-Lys)-NH.sub.2 110* 95
Ac-Arg-cyclo(Asp-Thr-D-Phe-Arg-Trp-Lys)-NH.sub.2 ND 96
Ac-Arg-cyclo(Lys-His-D-Phe-Arg-Trp-Asp)-NH.sub.2 1* 97
Ac-Arg-cyclo(Asp-Phe(4-(C(.dbd.O)--NH.sub.2))-D-Phe-Arg-Trp-Lys)-OH
ND 98 Ac-D-Arg-cyclo(Orn-His-D-Phe-Arg-Trp-Glu)-NH.sub.2 4* 99
Arg-cyclo(Orn-His-D-Phe-Arg-Trp-Glu)-NH.sub.2 6 100
Ac-Arg-cyclo(Glu-Met(O.sub.2)-D-Phe-Arg-Trp-Orn)-NH.sub.2 8 101
Ac-Arg-cyclo(Glu-Hyp-D-Phe-Arg-Trp-Orn)-NH.sub.2 3 102
Ac-Arg-cyclo(Glu-Arg-D-Phe-Arg-Trp-Orn)-NH.sub.2 0.7 103
Ac-Arg-cyclo(Glu-Lys-D-Phe-Arg-Trp-Orn)-NH.sub.2 15 104
Ac-Arg-cyclo(Glu-Orn-D-Phe-Arg-Trp-Orn)-NH.sub.2 9* 105
Ac-Arg-cyclo(Glu-Cit-D-Phe-Arg-Trp-Orn)-NH.sub.2 6 106
Ac-Arg-cyclo(Asp-Gln-D-Phe-Arg-Trp-Lys)-NH-cyclopropyl 64* 107
Ac-Arg-cyclo(Glu-Gln-D-Phe-Arg-Trp-Orn)-NH-Et 34* 108
Ac-Arg-cyclo(Glu-Gln-D-Phe-Arg-Trp-Orn)-NH-cyclopropyl 26* 109
Ac-Arg-cyclo(Glu-His-D-Phe-Arg-Trp-Orn)-NH-Et 6* 110
Ac-Arg-cyclo(Glu-His-D-Phe-Arg-Trp-Orn)-NH-cyclopropyl 1* 111
Ac-Arg-cyclo(Glu-Met(.dbd.O)-D-Phe-Arg-Trp-Orn)-NH.sub.2 9 112
Ac-Arg-cyclo(Glu-Pro(4R-NH.sub.2)-D-Phe-Arg-Trp-Orn)-NH.sub.2 6 113
Ac-Arg-cyclo(Asp-Thr(Bzl)-D-Phe-Arg-Trp-Lys)-OH ND 114
Ac-Arg-cyclo(Asp-Dab(Acetyl)-D-Phe-Arg-Trp-Lys)-NH.sub.2 88* 115
Ac-Arg-cyclo(Asp-His-D-Phe-Cit-Trp-Lys)-NH.sub.2 87 116
Ac-Arg-cyclo(Asp-Lys-D-Phe-Cit-Trp-Lys)-NH.sub.2 873 117
Ac-Arg-cyclo(Asp-Ser-D-Phe-Cit-Trp-Lys)-NH.sub.2 1446 118
Ac-Arg-cyclo(Asp-Dap(betaPro)-D-Phe-Arg-Trp-Lys)-NH.sub.2 145* 119
Ac-Arg-cyclo(Orn-Dab-D-Phe-Arg-Trp-Glu)-NH.sub.2 3* 120
Ac-Arg-cyclo(Orn-Ala-D-Phe-Arg-Trp-Glu)-NH.sub.2 19* 121
Ac-Arg-cyclo(Lys-Dab-D-Phe-Arg-Trp-Asp)-NH.sub.2 0.4 122
Ac-Arg-cyclo(Asp-Dab-D-Phe-Arg-Trp-Lys)-NH-cyclopropyl 3
123 Ac-Arg-cyclo(Asp-Pro(4R-2-Cl-Bzl)-D-Phe-Arg-Trp-Lys)-NH.sub.2
8* 124
Ac-Arg-cyclo(Asp-Pro(4R-3-Cl-Bzl)-D-Phe-Arg-Trp-Lys)-NH.sub.2 19*
125 Ac-Arg-cyclo(Asp-Pro(4R-4-Cl-Bzl)-D-Phe-Arg-Trp-Lys)-NH.sub.2
24* 126 Ac-Arg-cyclo(hGlu-His-D-Phe-Arg-Trp-Dab)-NH.sub.2 1* 127
Ac-Arg-cyclo(hGlu-Dab-D-Phe-Arg-Trp-Dab)-NH.sub.2 2* 128
Ac-Arg-cyclo(Dab-His-D-Phe-Arg-Trp-hGlu)-NH.sub.2 2* 129
Ac-Arg-cyclo(Dab-Dab-D-Phe-Arg-Trp-hGlu)-NH.sub.2 4* 130
Ac-Arg-cyclo(Orn-Dab-D-Phe-Arg-Trp-Glu)-OH 25 131
Ac-Arg-cyclo(Lys-Dab-D-Phe-Arg-Trp-Asp)-OH 35 132
Ac-Lys-cyclo(Orn-His-D-Phe-Arg-Trp-Glu)-NH.sub.2 3* 133
Ac-D-Lys-cyclo(Orn-His-D-Phe-Arg-Trp-Glu)-NH.sub.2 8* 134
Arg-cyclo(Asp-Ser(Bzl)-D-Phe-Arg-Trp-Lys)-OH 30 135
Ac-Lys-cyclo(Asp-His-D-Phe-Arg-Trp-Lys)-NH.sub.2 9* 136
Ac-Arg-cyclo(Dab-Dab-D-Phe-Arg-Trp-hGlu)-OH 35 137
Cyclohexanoyl-Arg-cyclo(Asp-Dab-D-Phe-Arg-Trp-Lys)-NH.sub.2 0.4 138
Cyclopentylacetyl-Arg-cyclo(Asp-Dab-D-Phe-Arg-Trp-Lys)-NH.sub.2 0.8
139 Cyclohexylacetyl-Arg-cyclo(Asp-Dab-D-Phe-Arg-Trp-Lys)-NH.sub.2
1 140 Phenylacetyl-Arg-cyclo(Asp-Dab-D-Phe-Arg-Trp-Lys)-NH.sub.2 2
141 Ac-Cit-cyclo(Asp-Dab-D-Phe-Arg-Trp-Lys)-NH.sub.2 9 142
Ac-Gln-cyclo(Asp-Dab-D-Phe-Arg-Trp-Lys)-NH.sub.2 15 143
Ac-Arg-cyclo(Glu-Dab(Acetyl)-D-Phe-Arg-Trp-Orn)-NH.sub.2 13 144
Ac-Arg-cyclo(hGlu-Dab-D-Phe-Arg-Trp-Dab)-OH 45 145
Ac-Arg-cyclo(hGlu-Met(O.sub.2)-D-Phe-Arg-Trp-Dab)-NH.sub.2 4 146
Ac-Arg-cyclo(hGlu-Hyp-D-Phe-Arg-Trp-Dab)-NH.sub.2 3 147
Ac-Arg-cyclo(hGlu-Gln-D-Phe-Arg-Trp-Dab)-NH.sub.2 20 148
Ac-Arg-cyclo(Asp-Orn(Acetyl)-D-Phe-Arg-Trp-Lys)-NH.sub.2 573 149
Ac-Arg-cyclo(Glu-Orn(Acetyl)-D-Phe-Arg-Trp-Orn)-NH.sub.2 7 150
Ac-Arg-cyclo(Orn-Gln-D-Phe-Arg-Trp-Glu)-NH.sub.2 5 151
Ac-Arg-cyclo(Lys-Gln-D-Phe-Arg-Trp-Asp)-NH.sub.2 4 152
Ac-D-Arg-cyclo(Orn-Gln-D-Phe-Arg-Trp-Glu)-NH.sub.2 6 153
n-C.sub.4-H.sub.9-CO-cyclo(Glu-His-D-Phe-Arg-Dab)-Trp-NH.sub.2 365
154 Ac-Arg-cyclo(Orn-Asn-D-Phe-Arg-Trp-Glu)-NH.sub.2 13 155
Ac-Arg-cyclo(Lys-Asn-D-Phe-Arg-Trp-Asp)-NH.sub.2 5 156
Ac-D-Arg-cyclo(Orn-Asn-D-Phe-Arg-Trp-Glu)-NH.sub.2 28 157
Arg-cyclo(Orn-Asn-D-Phe-Arg-Trp-Glu)-NH.sub.2 42 158
Ac-Orn-cyclo(Orn-Asn-D-Phe-Arg-Trp-Glu)-NH.sub.2 50 159
Ac-Arg-cyclo(Dab-Gln-D-Phe-Arg-Trp-hGlu)-NH.sub.2 4 160
Ac-Arg-cyclo(hGlu-Asn-D-Phe-Arg-Trp-Dab)-NH.sub.2 8 161
Ac-Arg-cyclo(Glu-Asn-D-Phe-Arg-Trp-Orn)-NH.sub.2 6 162
Ac-Arg-cyclo(Lys-Asn-D-Phe-Arg-Nal 2-Asp)-NH.sub.2 5 163
Ac-Arg-cyclo(Asp-Dab-D-Phe-Arg-Trp-Lys)-NH.sub.2 4 164
Ac-Arg-cyclo(Asp-Dab-D-Phe-Arg-Trp-Lys)-OH 57 165
Ac-Arg-cyclo(Glu-Dab-D-Phe-Arg-Trp-Orn)-NH.sub.2 0.65 166
Ac-Arg-cyclo(Glu-Dab-D-Phe-Arg-Trp-Orn)-OH 8
[0156] 2. The peptides of Nos. 126 through 129 were tested in
functional assays. The peptide No. 126 was determined to be an
agonist at MC4r, with intrinsic activity of 101% at MC4r where
NDP-.alpha.-MSH is 100%, and with an EC.sub.50 of 0.047 nM (average
of three studies). The peptide No. 127 was determined to be an
agonist at MC4r, with intrinsic activity of 98% at MC4r where
NDP-.alpha.-MSH is 100%, and with an EC.sub.50 of 0.06 nM (average
of two studies). The peptide No. 128 was determined to be an
agonist at MC4r, with intrinsic activity of 95% at MC4r where
NDP-.alpha.-MSH is 100%, and with an EC.sub.50 of 0.073 nM (average
of three studies). The peptide No. 129 was determined to be an
agonist at MC4r, with intrinsic activity of 96% at MC4r where
NDP-.alpha.-MSH is 100%, and with an EC.sub.50 of 0.065 nM (average
of two studies). Thus for the four peptides in the series,
EC.sub.50 values were on the order of one and one-half-fold to
two-fold less that the Ki values.
[0157] 3. Peptide No. 1 was evaluated for binding against MC1r,
MC3r and MC4r in competitive studies using Eu-labeled
NDP-.alpha.-MSH, and was found to have a Ki value of 4 nM at MC4r
(average of six studies), a Ki value of 4 nM for MC1r (average of
four studies) and a Ki value of 103 nM for MC3r (average of five
studies). In competitive studies using [1125]-NDP-.alpha.-MSH,
peptide No. 1 was found to have a Ki value of 2 nM at MC4r (one
study), 25 nM at MC3r (one study) and 3 nM at MC1r (one study). In
functional studies, Peptide No. 1 was determined to be an agonist,
with intrinsic activity of 91% at MC4r where NDP-.alpha.-MSH is
100%, and with an EC.sub.50 of 1 nM (average of three studies).
[0158] In rat feeding studies, using bremelanotide (a non-specific
MC4r agonist of the formula
Ac-Nle-cyclo(Asp-His-D-Phe-Arg-Trp-Lys)-OH)) as a positive control,
peptide No. 1 was found to reduce food intake and decrease the rate
of change in body weight. Using the methods as described above,
groups of 8 rats each rats received 1 mg/kg bremelanotide, 0.3
mg/kg of peptide No. 1, 1 mg/kg of peptide No. 1 or vehicle
control. For the 0-2, 0-4, and 0-20 hour periods, the decrease in
food consumption in rats receiving either 0.3 or 1 mg/kg of peptide
No. 1 was statistically significant compared to control. The 0-20
hour percent change in body weight was also statistically
significant compared to control for the group receiving 1 mg/kg of
peptide No. 1.
[0159] 4. Peptide No. 16 was evaluated for binding against MC1r,
MC3r and MC4r in competitive studies using Eu-labeled
NDP-.alpha.-MSH, and was found to have a Ki value of 25 nM at MC4r
(average of two studies), a Ki value of 323 nM for MC1r (one study)
and a Ki value of 1055 nM for MC3r (one study). In functional
studies, Peptide No. 16 was determined to be a partial agonist,
with intrinsic activity of 42% at MC4r where NDP-.alpha.-MSH is
100%, and with an EC.sub.50 of 40 nM (average of five studies).
[0160] In rat feeding studies, using bremelanotide as a positive
control, peptide No. 16 was found to reduce food intake and
decrease the rate of change in body weight. Using the methods as
described above, groups of 8 rats each rats received 1 mg/kg
bremelanotide, 0.3 mg/kg of peptide No. 16, 1 mg/kg of peptide No.
16 or vehicle control. For the 0-2, 0-4, and 0-20 hour periods, the
decrease in food consumption in rats receiving 1 mg/kg of peptide
No. 16 was statistically significant compared to control, and for
the 0-2 and 0-4 hour periods, the decrease in food consumption in
rats receiving 0.3 mg/kg of peptide No. 16 was statistically
significant compared to control. The 0-20 hour percent change in
body weight was also statistically significant compared to control
for the group receiving 1 mg/kg of peptide No. 16.
[0161] 5. Peptide No. 32 was evaluated for binding against MC1r and
MC4r in competitive studies using Eu-labeled NDP-.alpha.-MSH, and
was found to have a Ki value of 24 nM at MC4r (average of six
studies) and a Ki value of 673 nM for MC1r (average of three
studies). In competitive studies using [I.sup.125]-NDP-.alpha.-MSH,
peptide No. 32 was found to have a Ki value of 13 nM at MC4r (two
studies), 340 nM at MC3r (one study) and 133 nM at MC1r (two
studies). In functional studies, Peptide No. 32 was determined to
be an agonist, with intrinsic activity of 98% at MC4r where
NDP-.alpha.-MSH is 100%, and with an EC.sub.50 of 17 nM (average of
eight studies).
[0162] 6. Certain peptides were tested for binding selectivity (Ki)
to MC1r and MC4r, and also for functional selectivity (EC.sub.50
and intrinsic activity, referred to as "E.sub.max" below) to both
Mc1r and MC4r, with the results as shown below:
TABLE-US-00003 hMC4r hMC1r hMC4r hMC4r hMC1r hMC1r Peptide (Ki)
(Ki) EC.sub.50 E.sub.max EC.sub.50 E.sub.max No. (nM) (nM) (nM) (%)
(nM) (%) 71 45 3350 2 89 60 88 150 5 130 0.3 87 9 89 152 6 120 0.5
89 18 85 154 13 688 0.625 87 25 87 155 5 240 0.189 97 14 91 156 28
840 2 89 80 81 159 4 133 0.825 99 26 98 160 8 295 0.245 98 18 95
161 6 155 0.413 91 13 85 162 5 255 0.85 82 22 89
[0163] Thus while each of the foregoing peptides is an agonist at
both MC1r and MC4r (defined as 70% or greater intrinsic activity),
in all instances both binding selectivity and functional
selectivity was at least twenty times more specific at MC4r than at
MC1r.
[0164] 7. Peptide Nos. 154 and 155 were evaluated in
pharmacokinetic studies, and in a rat model with subcutaneous
injection of the formulation as in Section 9.7, each had a terminal
half-life of approximately 0.7 hours.
[0165] 8. Peptide No. 154 was evaluated with GLP-1 (Alfa Aesar,
catalog J66197, GLP-1 (7-36) amide, human) to determine effect on
body weight and feed intake in diet induced obese mice. Diet
induced obese mice were divided into groups and subcutaneously
implanted with Alzet continuous infusion pumps containing one of
the following formulations: [0166] Group 1: Vehicle [0167] Group 2:
30 mg/kg/day Peptide No. 154 (MC4r 154) by bolus administration
[0168] Group 3: 0.1 mg/kg/day GLP-1 by continuous infusion [0169]
Group 4: 30 mg/kg/day MC4r 154 by bolus administration and 0.1
mg/kg/day GLP-1 by continuous infusion
[0170] Groups 1, 2 and 4 were provided ad libitum with a 45% high
fat rodent diet. Animals in Group 3 were pair-fed to Group 2. The
amount of food consumed in Group 2 was the amount of food given to
Group 3 in the next 24 hour period. For group 3 (pair-fed group),
the pump was implanted 1 day later than other groups to account for
pair-feeding. On day -2 and day 6, animals from Groups 1, 2, and 4
were fasted overnight and baseline and terminal glucose tolerance
tests (GTT) was conducted. GTT and insulin levels in Group 3 were
conducted one day later. Body weight and food intake were measured
each day. To conduct the GTT, on study day -2 (-1 for Group 3)
(baseline test) and day 6 (7 for Group 3) (termination test), 1
g/kg glucose was orally administered. Glucose levels were taken
prior to glucose administration and 15, 30, 60, 90 and 120 minutes
post-glucose administration.
[0171] FIG. 1 shows the percent change in the area under the curve
(AUC) for the GTT from the baseline test to the termination test.
Data is expressed as percent change at the termination test,
.+-.standard deviation (SD) from the baseline test. This was
calculated as:
(AUC.sub.day 7-AUC.sub.day-2)/AUC.sub.day 7
In FIG. 1, diet induced obese mice were administered MC4r 154 (30
mg/kg/day) either with or without 0.1 mg/kg/day GLP-1, with one
group pair fed to the MC4r 154 group but administered only GLP-1
("Pair-Fed to MC4r 154+GLP-1").
[0172] FIG. 2 shows the body weight change from study day 0 (day
prior to initiation of administration of formulations) through
study day 5. Data is shown as the average of body weight change in
grams .+-.standard error of the mean (SEM) from study day 0. As is
shown in FIG. 2, the greatest weight loss was seen in Group 4,
which was concurrently administered both MC4r 154 and GLP-1.
Virtually no change in weight was seen in Group 3, which was
administered only GLP-1 and was pair-fed with Group 2.
[0173] 9. MC4r 154 was evaluated with GLP-1 to determine effect on
body weight and feed intake in diet induced obese Sprague-Dawley
rats. Rats were acquired at 17 weeks and pre-conditioned for 5
weeks on high fat diet prior to dosing. No sharp increase in weight
was noted, likely due to late onset of the high fat diet. At study
day -2, baseline GTT and insulin levels were assessed on 16-hour
fasted animals. Animals were then permitted a 2-day recovery
period, and on study day 0, animals were randomized into treatment
groups by body weight and Alzet continuous infusion pumps
containing either vehicle or GLP-1 (for dosing at either 10 or 100
.mu.g/kg/day) were implanted. Beginning on study day 1, body weight
and feed weights were taken in the mornings. Beginning on study day
4, twice daily (b.i.d.) subcutaneous dosing of saline was initiated
for all groups other than the treatment naive group to acclimate
the animals to handling and subcutaneous dosing. Beginning on study
day 7, and through study day 11, all groups other than treatment
naive, vehicle control and GLP-1 only groups were administered a
bolus injection of either 0.1 or 1 mg/kg of Peptide No. 154 b.i.d.,
with the final dose of Peptide No. 154 on study day 12, one hour
prior to GTT and insulin testing. Animals were fasted for the 16
hours prior to the GTT.
[0174] FIG. 3 shows the percent change for the glucose test from
the baseline test (study day -2) to the termination test (study day
12, after 16 hour fast and prior to GTT testing). Data is shown as
the percent change from study day -2 baseline test .+-.SEM. As show
in FIG. 3, there was little or no change in glucose levels in
groups receiving only GLP-1. The most pronounced decrease in fasted
plasma glucose was seen in animals receiving 100 .mu.g/kg/day GLP-1
and a bolus injection of 0.1 mg/kg Peptide No. 154 b.i.d., and in
animals receiving a bolus injection of 1.0 mg/kg Peptide No. 154
b.i.d, either alone or with either 10 or 100 .mu.g/kg/day
GLP-1.
[0175] FIG. 4 compares the fasted insulin levels at study day -2
and study day 12, with insulin measured after 16 hour fast and
prior to GTT testing. Data is shown as ng/mL insulin .+-.SEM. As
can be seen, the most pronounced decrease in insulin levels from
study day -2 to study day 12 (p-value <0.05, as shown by the
asterisk) was seen in groups receiving GLP-1 at either 10 or 100
.mu.g/kg/day in combination with the bolus injection of 1.0 mg/kg
Peptide No. 154 b.i.d.
[0176] 10. In other studies, it was observed that even in the
absence of a GLP-1 receptor agonist that subcutaneous bolus
injection of Peptide No. 154 resulted in greater weight loss, and
continued weight loss over time, compared to continuous infusion of
Peptide No. 154. Weight loss was compared between Group 2 in
Example 8 above, in which DIO mice received continuous infusion of
30 mg/kg/day, and a study in which DIO mice received 3 mg/kg of
Peptide No. 154 in a single daily bolus dose. As shown in FIG. 2,
with continuous infusion of 30 mg/kg/day animals lost weight only
for the first two days, and reached a maximum weight loss of 1.76
g.+-.0.34 SEM compared to baseline (study day 0). By contrast,
animals receiving a single daily subcutaneous dose of 3 mg/kg
continuously lost weight over 5 days, and reached a maximum weight
loss on day 5 of 4.2 g.+-.0.6 SEM compared to baseline (study day
0).
[0177] 11. Peptide No. 154 was evaluated with GLP-1 (GenScript) to
determine effect on body weight and feed intake in diet induced
obese rats. Rats were implanted with an Azlet pump 2 ML2 at day 0,
and received continuous infusion of GLP-1 (at a concentration of
1.5 mg/mL for the 300 .mu.g/kg/day dose and a concentration of 5
mg/mL for the 1000 .mu.g/kg/day dose) or vehicle (3.2% mannitol and
50 mM Tris in deionized water, pH 7.4) on study days 0 through 10,
and twice daily subcutaneous injection of Peptide No. 154 at a dose
of 0.3, 1.0 or 3.0 mg/kg or vehicle in a volume of 1.2 mL vehicle
on days 5 through 10. The rats were divided into the following
groups [0178] Group 1: Sham [0179] Group 2: Vehicle [0180] Group 3:
1000 .mu.g/kg/day continuous infusion GLP-1 and 0.3 mg/kg Peptide
No. 154 by bolus subcutaneous administration twice daily [0181]
Group 4: 1000 .mu.g/kg/day continuous infusion GLP-1 and 1.0 mg/kg
Peptide No. 154 by bolus subcutaneous administration twice daily
[0182] Group 5: 1000 .mu.g/kg/day continuous infusion GLP-1 and 3.0
mg/kg Peptide No. 154 by bolus subcutaneous administration twice
daily [0183] Group 6: 300 .mu.g/kg/day continuous infusion GLP-1
and 0.3 mg/kg Peptide No. 154 by bolus subcutaneous administration
twice daily [0184] Group 7: 300 .mu.g/kg/day continuous infusion
GLP-1 and 1.0 mg/kg Peptide No. 154 by bolus subcutaneous
administration twice daily [0185] Group 8: 300 .mu.g/kg/day
continuous infusion GLP-1 and 3.0 mg/kg Peptide No. 154 by bolus
subcutaneous administration twice daily [0186] Group 9: 1000
.mu.g/kg/day continuous infusion GLP-1 [0187] Group 10: 300
.mu.g/kg/day continuous infusion GLP-1 [0188] Group 11: 0.3 mg/kg
Peptide No. 154 by bolus subcutaneous administration twice daily
[0189] Group 12: 1.0 mg/kg Peptide No. 154 by bolus subcutaneous
administration twice daily [0190] Group 13: 3.0 mg/kg Peptide No.
154 by bolus subcutaneous administration twice daily All data was
calculated as means.+-.SEM. Each treatment group was compared to
the vehicle group using t-test, one-way ANOVA or two-way ANOVA
followed by Dunnett's multiple comparisons (GraphPad). A p value
<0.05 was considered to represent a significant difference.
[0191] Individual body weights were measured on study days 1-9.
Percent change in body weight is calculating according to this
formula: 100*(value-baseline)/baseline, where value is the weight
of the animal on study day in question and baseline is the weight
of the same animal on study day -1. Compared with vehicle,
subcutaneous dosing of Peptide No. 154 decreased animal's body
weight significantly. Combination of continuous infusion of GLP-1
at dose of 1000 .mu.g/kg/day with Peptide No. 154 showed more
decrease in body weight.
[0192] On days 1-4, feeds were weighed at 0, and 24 hours after
lights off. On days 5-9, feeds were weighed at 0, 2, 6 and 24 hours
after lights off. Compared with vehicle, there was no significant
difference on feed intake with continuous infusion of GLP-1. Food
intake decreased starting on day 5 following subcutaneous dosing of
Peptide No. 154 in both groups receiving only Peptide No. 154 and
groups receiving a combination of GLP-1 and Peptide No. 154.
Appetites in groups receiving only Peptide No. 154 recovered on day
9, but remained decreased in groups receiving a combination of
GLP-1 and Peptide No. 154. At 2 and 6 hours post lights off, both
groups receiving only Peptide No. 154 and groups receiving a
combination of GLP-1 and Peptide No. 154 showed significant
decreased feed intakes compared with the vehicle group.
[0193] FIG. 6 shows average body weight change in percent of
baseline in DIO rats from study day 0 through study day 9 in DIO
rats administered vehicle, 1000 .mu.g/kg/day of GLP-1, 3 mg/kg MC4r
154, or a combination of MC4r 154 and GLP-1. GLP-1 was administered
via subcutaneously implanted osmotic pump on study days 0 to 10.
MC4r 154 was administered twice daily via subcutaneous injection on
study days 5 through 10. Body weight was measured daily prior to
beginning of the feeding cycle and prior to the first dose of MC4r
154.
[0194] FIG. 7 shows average body weight change in percent of
baseline in DIO rats from study day 0 through study day 9 in DIO
rats administered vehicle, 1000 .mu.g/kg/day of GLP-1, 0.3 mg/kg
MC4r 154, or a combination of MC4r 154 and GLP-1. GLP-1 was
administered via subcutaneously implanted osmotic pump on study
days 0 to 10. MC4r 154 was administered twice daily via
subcutaneous injection on study days 5 through 10. Body weight was
measured daily prior to beginning of the feeding cycle and prior to
the first dose of MC4r 154.
[0195] FIG. 8 shows average body weight change in percent of
baseline in DIO rats from study day 0 through study day 9 in DIO
rats administered vehicle, 300 .mu.g/kg/day of GLP-1, 1 mg/kg MC4r
154, or a combination of MC4r 154 and GLP-1. GLP-1 was administered
via subcutaneously implanted osmotic pump on study days 0 to 10.
MC4r 154 was administered twice daily via subcutaneous injection on
study days 5 through 10. Body weight was measured daily prior to
beginning of the feeding cycle and prior to the first dose of MC4r
154.
[0196] 12. In the study of Example 11 above, on day -2 (baseline)
and day 10 (termination), glucose was administered at 1 g/kg orally
to each animal. Glucose levels were taken on baseline (prior to
glucose) and 15, 30, 60, 90 and 120 minutes post glucose
administration. As shown in FIG. 5, after 10 days of treatment,
compared with baseline level, only groups receiving a combination
of GLP-1 and Peptide No. 154 showed a significant inhibition of
blood glucose level (p<0.01). FIG. 5 shows percent change in the
AUC for a glucose tolerance test from the baseline test to the
termination test with DIO rats administered vehicle, MC4r 154, or a
combination of MC4r 154 and GLP-1. GLP-1 was administered via
subcutaneously implanted osmotic pump on study days 0 to 10. MC4r
154 was administered twice daily via subcutaneous injection on
study days 5 through 10.
[0197] 13. Formulations are made consisting of the following active
ingredients per dose:
TABLE-US-00004 Formulation GLP- 1 Agonist MC4r Agonist 13.1 0.6 mg
liraglutide 1.0 mg Peptide 154 13.2 0.6 mg liraglutide 1.75 mg
Peptide 154 13.3 0.6 mg liraglutide 5.0 mg Peptide 154 13.4 0.6 mg
liraglutide 10.0 mg Peptide 154 13.5 1.5 mg liraglutide 1.0 mg
Peptide 154 13.6 1.5 mg liraglutide 1.75 mg Peptide 154 13.7 1.5 mg
liraglutide 5.0 mg Peptide 154 13.8 1.5 mg liraglutide 10.0 mg
Peptide 154
The exenatide and Peptide 154 are free-base and anhydrous weights.
Each dose unit is supplied for subcutaneous injection in a sterile,
preserve isotonic solution in a volume of between 0.3 and 0.5 mL,
which also optionally includes metacresol as an antimicrobial
preservative, mannitol as a tonicity-adjusting agent, and glacial
acetic acid and sodium acetate trihydrate in water for injection as
a buffering solution at pH 4.5.
[0198] Any of the formulations 13.1 through 13.8 are administered
to patients in need of treatment of obesity or to induce weight
loss, or to patients in need of improved glycemic control, by once
daily subcutaneous injection.
[0199] 14. A formulation is made consisting of the following active
ingredients per dose:
TABLE-US-00005 Formulation GLP-1 Agonist MC4r Agonist 14.1 5 .mu.g
extenatide 1.0 mg Peptide 154 14.2 5 .mu.g extenatide 1.75 mg
Peptide 154 14.3 5 .mu.g extenatide 2.5 mg Peptide 154 14.4 5 .mu.g
extenatide 150 mg Peptide 154 14.5 10 .mu.g extenatide 1.0 mg
Peptide 154 14.6 10 .mu.g extenatide 1.75 mg Peptide 154 14.7 10
.mu.g extenatide 2.5 mg Peptide 154 14.8 10 .mu.g extenatide 5.0 mg
Peptide 154 14.9 20 .mu.g extenatide 1.0 mg Peptide 154 14.10 20
.mu.g extenatide 1.75 mg Peptide 154 14.11 20 .mu.g extenatide 2.5
mg Peptide 154 14.12 20 .mu.g extenatide 5.0 mg Peptide 154
The liraglutide and Peptide 154 are free-base and anhydrous
weights. Each dose unit is supplied for subcutaneous injection in a
sterile, preserve isotonic solution in a volume of between 0.3 and
0.5 mL, which optionally also includes disodium phosphate
dihydrate, propylene glycol, phenol, and water for injection.
[0200] Any of the formulations 14.1 through 14.12 are administered
to patients in need of treatment of obesity or to induce weight
loss, or to patients in need of improved glycemic control, by twice
daily subcutaneous injection, preferably at least one hour prior to
breakfast and dinner.
[0201] Although the invention has been described in detail with
particular reference to these preferred embodiments, other
embodiments can achieve the same results. Variations and
modifications of the present invention will be obvious to those
skilled in the art and it is intended to cover all such
modifications and equivalents. The entire disclosures of all
references, applications, patents, and publications cited above are
hereby incorporated by reference.
Sequence CWU 1
1
3130PRTHomo sapiensMOD_RES(30)..(30)AMIDATION 1His Ala Glx Gly Thr
Phe Thr Ser Asx Val Ser Ser Tyr Leu Glx Gly1 5 10 15Glx Ala Ala Lys
Glx Phe Ile Ala Trp Leu Val Lys Gly Arg 20 25 30213PRTArtificial
SequenceSynthetic melanocortin binding peptide derived from human
alpha-MSHMOD_RES(1)..(1)ACETYLATIONMOD_RES(13)..(13)AMIDATION 2Ser
Tyr Ser Met Glu His Phe Arg Trp Gly Lys Pro Val1 5
1034PRTArtificial SequenceSynthetic melanocortin binding peptide
derived from human alpha-MSH 3His Phe Arg Trp1
* * * * *