U.S. patent application number 12/869917 was filed with the patent office on 2010-12-23 for therapeutic and prophylactic methods for neuromuscular disorders.
This patent application is currently assigned to WYETH LLC. Invention is credited to XIANGPING LI, LISA-ANNE WHITTEMORE.
Application Number | 20100322942 12/869917 |
Document ID | / |
Family ID | 33511616 |
Filed Date | 2010-12-23 |
United States Patent
Application |
20100322942 |
Kind Code |
A1 |
WHITTEMORE; LISA-ANNE ; et
al. |
December 23, 2010 |
THERAPEUTIC AND PROPHYLACTIC METHODS FOR NEUROMUSCULAR
DISORDERS
Abstract
The disclosure provides methods for treating neuromuscular
disorders in mammals. The disclosed methods include administering
therapeutically effective amounts of a GDF-8 inhibitor and a
corticosteroid to a subject susceptible to, or having, a
neuromuscular disorder, so as to maintain desirable levels of
muscle function.
Inventors: |
WHITTEMORE; LISA-ANNE; (EAST
WALPOLE, MA) ; LI; XIANGPING; (WAYLAND, MA) |
Correspondence
Address: |
WYETH LLC/FINNEGAN HENDERSON, LLP
901 NEW YORK AVENUE, NW
WASHINGTON
DC
20001-4413
US
|
Assignee: |
WYETH LLC
MADISON
NJ
|
Family ID: |
33511616 |
Appl. No.: |
12/869917 |
Filed: |
August 27, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10858353 |
Jun 1, 2004 |
7785587 |
|
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12869917 |
|
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60474603 |
Jun 2, 2003 |
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Current U.S.
Class: |
424/142.1 ;
424/145.1; 424/158.1; 424/172.1; 514/1.1; 514/171; 514/9.7 |
Current CPC
Class: |
A61P 29/00 20180101;
A61P 17/00 20180101; A61P 19/02 20180101; A61K 39/39533 20130101;
A61P 25/00 20180101; A61P 21/00 20180101; A61K 2039/505 20130101;
A61P 37/02 20180101; A61K 39/39533 20130101; A61P 43/00 20180101;
A61P 3/14 20180101; A61P 37/06 20180101; A61K 2300/00 20130101;
A61P 9/04 20180101; A61P 11/00 20180101; A61P 25/02 20180101; A61P
21/04 20180101; A61P 37/08 20180101; A61P 17/06 20180101; C07K
16/24 20130101 |
Class at
Publication: |
424/142.1 ;
514/171; 424/158.1; 424/172.1; 514/9.7; 424/145.1; 514/1.1 |
International
Class: |
A61K 39/395 20060101
A61K039/395; A61K 31/56 20060101 A61K031/56; A61K 31/573 20060101
A61K031/573; A61K 38/22 20060101 A61K038/22; A61K 38/02 20060101
A61K038/02; A61P 21/00 20060101 A61P021/00; A61P 25/00 20060101
A61P025/00 |
Claims
1. A method of treating a mammal with a decrease of muscle
function, comprising administering to the mammal a therapeutically
effective amount of at least one GDF-8 inhibitor and a
therapeutically effective amount of at least one corticosteroid in
the amounts and for a period of time sufficient to treat decrease
of muscle function.
2. The method of claim 1, wherein the muscle function of at least
one muscle is evaluated by at least one parameter chosen from
muscle mass, muscle contraction force, serum CK concentration, or
muscle morphology.
3. The method of claim 1, wherein the muscle whose function is
treated is chosen from at least one of gastrocnemius, tibialis
anterior, quadriceps, extensor digitorum longus, cardiac muscle, or
diaphragm muscle.
4. The method of claim 1, wherein treating said mammal results in
increased body weight of said mammal.
5. The method of claim 1, wherein treating said mammal results in
increased grip strength.
6. The method of claim 1, wherein the mammal is human.
7. The method of claim 1, wherein the corticosteroid is chosen from
at least one of: (a) at least one of beclomethasone dipropionate,
budesonide, cortisol, dexamethasone, fluticason propionate,
mometasone furoate, prednisone, or triamcinolone acetonide; (b) a
derivative of at least one of beclomethasone dipropionate,
budesonide, cortisol, dexamethasone, fluticason propionate,
mometasone furoate, prednisone, or triamcinolone acetonide; or (c)
a pharmaceutically acceptable salt of at least one of
beclomethasone dipropionate, budesonide, cortisol, dexamethasone,
fluticason propionate, mometasone furoate, prednisone, or
triamcinolone acetoniden.
8. The method of claim 1, wherein the corticosteroid is prednisone
or prednisolone.
9. The method of claim 1, wherein the corticosteroid is
administered at a dose between 0.1 and 2.0 mg/kg/day.
10. The method of claim 1, wherein the corticosteroid is
administered orally.
11. The method of claim 1, wherein the method results in treating
of cardiomyopathy of said mammal.
12. The method of claim 1, wherein the administration of GDF-8
inhibitor and corticosteroid is concurrent.
13. The method of claim 1, wherein the administration of GDF-8
inhibitor and corticosteroid is consecutive.
14. The method of claim 1, wherein the GDF-8 inhibitor is a small
molecule inhibitor.
15. The method of claim 1, wherein the GDF-8 inhibitor is chosen
from an antibody to GDF-8, an antibody to a GDF-8 receptor, a
soluble GDF-8 receptor, a GDF-8 propeptide, follistatin, or a
follistatin-domain-containing protein.
16. The method of claim 15, wherein the antibody to GDF-8 is chosen
from JA-16, Myo29, Myo28, or Myo22.
17. The method of claim 15, wherein the GDF-8 propeptide is mutated
at an aspartate residue.
18. The method of claim 15, wherein the GDF-8 propeptide is joined
to the Fc portion of an immunoglobulin.
19. The method of claim 15, wherein the GDF-8 receptor is
ActRIIB.
20. The method of claim 15, wherein the GDF-8 receptor is joined to
the Fc portion of an immunoglobulin.
21. The method of claim 15, wherein the GDF-8 inhibitor is
follistatin.
22. The method of claim 15, wherein the
follistatin-domain-containing protein is GASP-1.
23. A method of treating muscle weakness, comprising administering
to a mammal a therapeutically effective amount of at least one
GDF-8 inhibitor and a therapeutically effective amount of at least
one corticosteroid in the amounts and for a period of time
sufficient to treat loss of muscle strength.
24. The method of claim 23, wherein the mammal is human.
25. The method of claim 23, wherein the corticosteroid is chosen
from at least one of: (a) at least one of beclomethasone
dipropionate, budesonide, cortisol, dexamethasone, fluticason
propionate, mometasone furoate, prednisone, or triamcinolone
acetonide; (b) a derivative of at least one of beclomethasone
dipropionate, budesonide, cortisol, dexamethasone, fluticason
propionate, mometasone furoate, prednisone, or triamcinolone
acetonide; or (c) a pharmaceutically acceptable salt of at least
one of beclomethasone dipropionate, budesonide, cortisol,
dexamethasone, fluticason propionate, mometasone furoate,
prednisone, or triamcinolone acetoniden.
26. The method of claim 23, wherein the corticosteroid is
prednisone or prednisolone.
27. The method of claim 23, wherein the corticosteroid is
administered at a dose between 0.1 and 2.0 mg/kg/day.
28. The method of claim 23, wherein the corticosteroid is
administered orally.
29. The method of claim 23, wherein the GDF-8 inhibitor is a small
molecule inhibitor.
30. The method of claim 23, wherein the GDF-8 inhibitor is chosen
from an antibody to GDF-8, an antibody to a GDF-8 receptor, a
soluble GDF-8 receptor, a GDF-8 propeptide, follistatin, or a
follistatin-domain-containing protein.
31. The method of claim 30, wherein the antibody to GDF-8 is chosen
from JA-16, Myo29, Myo28, or Myo22.
32. The method of claim 30, wherein the GDF-8 propeptide is mutated
at an aspartate residue.
33. The method of claim 30, wherein the GDF-8 propeptide is joined
to the Fc portion of an immunoglobulin.
34. The method of claim 30, wherein the GDF-8 receptor is
ActRIIB.
35. The method of claim 30, wherein the GDF-8 receptor is joined to
the Fc portion of an immunoglobulin.
36. The method of claim 30, wherein the GDF-8 inhibitor is
follistatin.
37. The method of claim 30, wherein the
follistatin-domain-containing protein is GASP-1.
38. A method of treating corticosteroid-induced muscle atrophy,
comprising administering to a mammal a therapeutically effective
amount of at least one GDF-8 inhibitor sufficient to treat the
corticosteroid-induced muscle atrophy.
39. The method of claim 38, wherein the mammal is human.
40. The method of claim 38, wherein the corticosteroid is chosen
from at least one of: (a) at least one of beclomethasone
dipropionate, budesonide, cortisol, dexamethasone, fluticason
propionate, mometasone furoate, prednisone, or triamcinolone
acetonide; (b) a derivative of at least one of beclomethasone
dipropionate, budesonide, cortisol, dexamethasone, fluticason
propionate, mometasone furoate, prednisone, or triamcinolone
acetonide; or (c) a pharmaceutically acceptable salt of at least
one of beclomethasone dipropionate, budesonide, cortisol,
dexamethasone, fluticason propionate, mometasone furoate,
prednisone, or triamcinolone acetoniden.
41. The method of claim 38, wherein the corticosteroid is
prednisone or prednisolone.
42. The method of claim 38, wherein the corticosteroid is
administered at a dose between 0.1 and 2.0 mg/kg/day.
43. The method of claim 38, wherein the corticosteroid is
administered orally.
44. The method of claim 38, wherein the GDF-8 inhibitor is a small
molecule inhibitor.
45. The method of claim 38, wherein the GDF-8 inhibitor is chosen
from an antibody to GDF-8, an antibody to a GDF-8 receptor, a
soluble GDF-8 receptor, a GDF-8 propeptide, follistatin, or a
follistatin-domain-containing protein.
46. The method of claim 45, wherein the antibody to GDF-8 is chosen
from JA-16, Myo29, Myo28, or Myo22.
47. The method of claim 45, wherein the GDF-8 propeptide is mutated
at an aspartate residue.
48. The method of claim 45, wherein the GDF-8 propeptide is joined
to the Fc portion of an immunoglobulin.
49. The method of claim 45, wherein the GDF-8 receptor is
ActRIIB.
50. The method of claim 45, wherein the GDF-8 receptor is joined to
the Fc portion of an immunoglobulin.
51. The method of claim 45, wherein the GDF-8 inhibitor is
follistatin.
52. The method of claim 45, wherein the
follistatin-domain-containing protein is GASP-1.
53. A method of treating a neuromuscular disorder, comprising
administering to a mammal having or at risk of the neuromuscular
disorder a therapeutically effective amount of at least one GDF-8
inhibitor and a therapeutically effective amount of at least one
corticosteroid in the amounts and for a period of time sufficient
to treat the neuromuscular disorder.
54. The method of claim 53, wherein the neuromuscular disorder is a
muscular dystrophy.
55. The method of claim 54, wherein the muscular dystrophy is
Duchenne muscular dystrophy.
56. The method of claim 54, wherein the muscular dystrophy is
Becker muscular dystrophy.
57. The method of claim 53, wherein the mammal is human.
58. The method of claim 53, wherein the corticosteroid is chosen
from at least one of: (a) at least one of beclomethasone
dipropionate, budesonide, cortisol, dexamethasone, fluticason
propionate, mometasone furoate, prednisone, or triamcinolone
acetonide; (b) a derivative of at least one of beclomethasone
dipropionate, budesonide, cortisol, dexamethasone, fluticason
propionate, mometasone furoate, prednisone, or triamcinolone
acetonide; or (c) a pharmaceutically acceptable salt of at least
one of beclomethasone dipropionate, budesonide, cortisol,
dexamethasone, fluticason propionate, mometasone furoate,
prednisone, or triamcinolone acetoniden.
59. The method of claim 53, wherein the corticosteroid is
prednisone or prednisolone.
60. The method of claim 53, wherein the corticosteroid is
administered at a dose between 0.1 and 2.0 mg/kg/day.
61. The method of claim 53, wherein the corticosteroid is
administered orally.
62. The method of claim 53, wherein the GDF-8 inhibitor is a small
molecule inhibitor.
63. The method of claim 53, wherein the GDF-8 inhibitor is chosen
from an antibody to GDF-8, an antibody to a GDF-8 receptor, a
soluble GDF-8 receptor, a GDF-8 propeptide, follistatin, or a
follistatin-domain-containing protein.
64. The method of claim 63, wherein the antibody to GDF-8 is chosen
from JA-16, Myo29, Myo28, or Myo22.
65. The method of claim 63, wherein the GDF-8 propeptide is mutated
at an aspartate residue.
66. The method of claim 63, wherein the GDF-8 propeptide is joined
to the Fc portion of an immunoglobulin.
67. The method of claim 63, wherein the GDF-8 receptor is
ActRIIB.
68. The method of claim 63, wherein the GDF-8 receptor is joined to
the Fc portion of an immunoglobulin.
69. The method of claim 63, wherein the GDF-8 inhibitor is
follistatin.
70. The method of claim 63, wherein the
follistatin-domain-containing protein is GASP-1.
71. The method of claim 63, wherein the method results in treating
of cardiomyopathy of said mammal.
72. The method of claim 63, wherein the administration of GDF-8
inhibitor and corticosteroid is concurrent.
73. The method of claim 63, wherein the administration of GDF-8
inhibitor and corticosteroid is consecutive.
Description
[0001] This is a continuation of application Ser. No. 10/858,353,
filed Jun. 1, 2004, which claims priority to U.S. provisional
application No. 60/474,603, filed on Jun. 2, 2003, all of which are
hereby incorporated by reference in their entirety.
FIELD OF THE INVENTION
[0002] The present invention relates to the field of clinical
pathophysiology, and more particularly to methods for treating
neuromuscular disorders, such as muscular dystrophies. The
invention also relates to pharmaceutical formulations containing
corticosteroids and inhibitors of growth and differentiation.
BACKGROUND OF THE INVENTION
[0003] Muscular dystrophies (MD) are progressive inherited
neuromuscular disorders that are characterized by muscle wasting
and weakness (Emery (2002) The Lancet, 359:687-695). Many forms of
muscular dystrophies are fatal and currently incurable.
[0004] Duchenne muscular dystrophy (DMD) is the most common
X-linked neuromuscular disease. The disease is caused by mutations
in the DMD gene coding for dystrophin. Alteration or absence of
this protein results in abnormal sarcolemmal membrane tearing. An
abnormal variation in diameter of muscle fibers (atrophic and
hypertrophic fibers) in proximal muscles and ongoing muscle damage
are hallmarks of the disease. Damaged muscle releases the
intracellular enzyme creatine kinase (CK). As a result, the serum
CK levels in DMD patients are characteristically high (up to 10
times the normal). The pathophysiologic cascade is compounded by
tissue inflammation, myofiber necrosis and replacement of muscle
with fibrofatty tissue.
[0005] Another allelic variant of the DMD gene causes a milder form
of MD known as Becker muscular dystrophy (BMD). BMD is clinically
similar to DMD but the onset of symptoms occurs later in life.
[0006] Many pharmacological agents have been tried in MD but none
has proved effective in arresting the course of the disease. The
current modality of treatment is still in the realm of physical
medicine and rehabilitation.
[0007] A number of trials using corticosteroids (e.g., prednisone
and/or its derivatives) have demonstrated improvement in
individuals with MD, particularly in the short-term. Although the
exact mechanism by which corticosteroids alleviate the disease
phenotype is unclear, corticosteroids are thought to act by
reducing inflammation, suppressing the immune system, improving
calcium homeostasis, upregulating expression of compensatory
proteins, and increasing myoblast proliferation (Khurana et al.
(2003) Nat. Rev. Drug Discovery 2:279-386). However,
corticosteroids administered over time can induce muscle atrophy,
which primarily affects proximal muscles--the very same muscles
that are affected in DMD and BMD. The corticosteroid-induced muscle
and other side effects may limit the long-term effectiveness of
corticosteroid therapy.
[0008] GDF-8 is a member of the TGF-.beta. superfamily and
functions as a negative regulator of muscle growth. Similarly to
other members of the superfamily, GDF-8 is synthesized as a
precursor molecule, but prior to secretion, it is cleaved into the
N-terminal inhibitory propeptide and C-terminal the active mature
GDF-8. Propeptide may remain bound to GDF-8 thereby inhibiting the
biological activity of mature GDF-8. Propeptide must dissociate
from the complex for GDF-8 to bind to activin type II receptor
(ActRIIB). Upon binding, ActRIIB initiates a signaling cascade,
ultimately leading to the inhibition of myoblast progression.
Antibody-mediated inhibition of GDF-8 in vivo has been shown to
significantly increase skeletal muscle size in normal adult mice
(Whittemore et al. (2003) BBRC, 300:965-971) and to alleviate the
dystrophic phenotype in the mdx mouse model of DMD (Bogdanovich et
al. (2002) Nature, 420(28):418-421).
SUMMARY OF THE INVENTION
[0009] It is one of the objects of the present invention to provide
methods and compositions for treating disorders characterized by or
associated with a risk of diminution of muscle function. Additional
objects of the invention will be set forth in part in the following
description, and in part will be understood from the description,
or may be learned by practice of the invention.
[0010] The present invention is based, in part, on the discovery
and demonstration that, in a mouse model of DMD, treatment by
administration of a neutralizing anti-GDF-8 antibody and prednisone
is more effective in increasing muscle mass and strength relative
to treatment with prednisone alone. The invention is further based,
in part, on the discovery and demonstration that administration of
anti-GDF-8 antibody with prednisone reduces prednisone-induced
muscle atrophy.
[0011] Accordingly, the present invention provides methods for
treating neuromuscular disorders in mammals. The disclosed methods
include administering to a subject susceptible to or having a
neuromuscular disorder therapeutically effective amounts of at
least one GDF-8 inhibitor and at least one corticosteroid so as to
maintain desirable levels of muscle integrity or function as
assessed by, for example, serum concentration of creatine kinase
(CK), muscle histology, tissue imaging, activities of daily living,
muscle strength and/or mass. The populations treated by the methods
of the invention include, but are not limited to, patients having
or at risk of developing muscular dystrophy such as, for example,
DMD or BMD, and subjects undergoing corticosteroid therapy for
these or other disorders.
[0012] The invention further provides methods of treating muscle
weakness and methods of treating corticosteroid-induced muscle
atrophy. The invention includes methods of treating
cardiomyopathy.
[0013] Methods of administration and compositions used in the
methods of the inventions are provided. In the disclosed methods, a
GDF-8 inhibitor and a corticosteroid are administered concurrently
or over alternating overlapping or non-overlapping intervals.
[0014] GDF-8 inhibitors, used in the methods of the present
invention, include, but are not limited to, antibodies to GDF-8;
antibodies to GDF-8 receptors; soluble GDF-8 receptors and
fragments thereof (e.g., ActRIIB fusion polypeptides as described
in U.S. patent application Ser. No. 10/689,677, including soluble
ActRIIB receptors in which ActRIIB is joined to the Fc portion of
an immunoglobulin); GDF-8 propeptide and modified forms thereof
(e.g., as described in WO 02/068650 or U.S. patent application Ser.
No. 10/071,499, now U.S. Pat. No. 7,202,210, including forms in
which GDF-8 propeptide is joined to the Fc portion of an
immunoglobulin and/or form in which GDF-8 is mutated at an
aspartate (asp) residue, e.g., asp-99 in murine GDF-8 propeptide
and asp-100 in human GDF-8 propeptide); a small molecule inhibitor
of GDF-8; follistatin (e.g., as described in U.S. Pat. No.
6,004,937) or follistatin-domain-containing proteins (e.g., GASP-1
or other proteins as described in U.S. patent application Ser. Nos.
10/369,736 and 10/369,738, now U.S. Pat. Nos. 7,192,717 and
7,572,763); and modulators of metalloprotease activity that affect
GDF-8 activation, as described in U.S. patent application Ser. No.
10/662,438.
[0015] In some embodiments, the GDF-8 inhibitor is a monoclonal
antibody or a fragment thereof that blocks GDF-8 binding to its
receptor. Nonlimiting illustrative embodiments include a nonhuman
monoclonal anti-GDF-8 antibody, e.g., murine monoclonal antibody
JA-16 (as described in U.S. patent application Ser. No. 10/253,532,
now U.S. Pat. No. 7,320,789; deposited with American Type Culture
Collection (ATCC), 10801 University Blvd., Manassas, Va., USA, on
Apr. 18, 2002, under the requirements of the Budapest Treaty and
accorded ATCC Deposit No. PTA-4236); derivatives thereof, e.g.,
humanized antibody; and fully human monoclonal anti-GDF-8
antibodies (e.g., Myo29, Myo28, and Myo22, as described in U.S.
patent application Ser. No. 10/688,925, now U.S. Pat. No.
7,261,893; ATCC Deposit Nos. PTA-4741, PTA-4740, and PTA-4739,
respectively) or derivatives thereof.
[0016] Corticosteroids, used in the method of the invention
include, but are not limited to, beclomethasone dipropionate,
budesonide, cortisol, dexamethasone, fluticason propionate,
mometasone furoate, prednisone, triamcinolone acetonide, and
derivatives thereof.
[0017] It is to be understood that both the foregoing general
description and the following detailed description are exemplary
and explanatory only and are not restrictive of the invention as
claimed.
BRIEF DESCRIPTION OF THE FIGURES
[0018] FIGS. 1A and 1B depict results of a histological analysis of
diaphragm muscle from mdx mice treated for four weeks with
anti-GDF-8 neutralizing antibody JA-16 (60 mg/kg, once weekly) and
prednisone (2 mg/kg, 3 times a week), prednisone alone, or vehicle
control alone. FIG. 1A shows severity of muscle fiber atrophy on a
0-4 scale at the end of the trial. FIG. 1B shows percentage of
affected (atrophied) muscle fibers at the end of the trial. Each
bar represents a single mouse.
DETAILED DESCRIPTION OF THE INVENTION
I. Definitions
[0019] In order that the present invention may be more readily
understood, certain terms are first defined. Additional definitions
are set forth throughout the detailed description.
[0020] The term "antibody," as used herein, refers to an
immunoglobulin or a part thereof and encompasses any polypeptide
comprising an antigen-binding site regardless of the source, method
of production, and other characteristics. As a non-limiting
example, the term "antibody" includes human, orangutan, mouse, rat,
goat, sheep, and chicken antibodies. The term includes but is not
limited to polyclonal, monoclonal, monospecific, polyspecific,
non-specific, humanized, single-chain, chimeric, synthetic,
recombinant, hybrid, mutated, and CDR-grafted antibodies. For the
purposes of the present invention, it also includes, unless
otherwise stated, antibody fragments such as Fab, F(ab')2, Fv,
scFv, Fd, dAb, and other antibody fragments that retain the
antigen-binding function.
[0021] Antibodies can be made, for example, via traditional
hybridoma techniques (Kohler and Milstein (1975) Nature, 256:
495-499), recombinant DNA methods (U.S. Pat. No. 4,816,567), or
phage display techniques using antibody libraries (Clackson et al.
(1991) Nature, 352: 624-628; Marks et al. (1991) J. Mol. Biol.,
222: 581-597). For various other antibody production techniques,
see Antibodies: A Laboratory Manual, eds. Harlow et al., Cold
Spring Harbor Laboratory, 1988.
[0022] The term "antigen-binding domain" refers to the part of an
antibody molecule that comprises the area specifically binding to
or complementary to a part or all of an antigen. Where an antigen
is large, an antibody may only bind to a particular part of the
antigen. The epitope or antigenic determinant is a portion of an
antigen molecule that is responsible for specific interactions with
the antigen-binding domain of an antibody. An antigen-binding
domain may be provided by one or more antibody variable domains
(e.g., a so-called Fd antibody fragment consisting of a VH domain).
An antigen-binding domain comprises an antibody light chain
variable region (VL) and an antibody heavy chain variable region
(VH).
[0023] The term "anti-GDF-8 antibody," or "antibody to GDF-8,"
refers to any antibody that specifically binds to at least one
epitope of GDF-8. The terms "GDF-8 receptor antibody" and "antibody
to a GDF-8 receptor" refer to any antibody that specifically binds
to at least one epitope of a GDF-8 receptor, such as ActRIIB. The
term "neutralizing antibody" refers to an antibody that is a GDF-8
inhibitor.
[0024] The term "specific interaction," or "specifically binds," or
the like, means that two molecules form a complex that is
relatively stable under physiologic conditions. The term is also
applicable where, e.g., an antigen-binding domain is specific for a
particular epitope, which may be present on a number of antigens.
Specific binding is characterized by a high affinity and a low to
moderate capacity. Nonspecific binding usually has a low affinity
with a moderate to high capacity. Typically, the binding is
considered specific when the affinity constant K.sub.a is higher
than 10.sup.6 M.sup.-1, than 10.sup.7 M.sup.-1, or preferably
higher than 10.sup.8 M.sup.-1. If necessary, non-specific binding
can be reduced without substantially affecting specific binding by
varying the binding conditions. Such conditions are known in the
art, and a skilled artisan using routine techniques can select
appropriate conditions. The conditions are usually defined in terms
of concentration of antibodies, ionic strength of the solution,
temperature, time allowed for binding, concentration of non-related
molecules (e.g., serum albumin, milk casein), etc.
[0025] The term "muscle function" refers to the ability of muscle
to perform a physiologic function, such as contraction as measured
by the amount of force generated during either twitch or tetanus.
Other methods for assessing muscle function are well known in the
art and include, but are not limited to, measurements of muscle
mass, grip strength, serum CK level, activities of daily living,
motion or strength tests, tissue histology (e.g., E&A staining,
or collagen III staining), or tissue imaging. Nonlimiting
illustrative methods for assessing muscle function are set forth in
the Examples.
[0026] The term "GDF-8" refers to a specific growth and
differentiation factor-8 and, where appropriate, factors that are
structurally or functionally related to GDF-8, for example, BMP-11
and other factors belonging to the TGF-.beta. superfamily. The term
refers to the full-length unprocessed precursor form of GDF-8 as
well as the mature and propeptide forms resulting from
post-translational cleavage. The term also refers to any fragments
and variants of GDF-8 that maintain at least some biological
activities associated with mature GDF-8, as discussed herein,
including sequences that have been modified. The present invention
relates to GDF-8 from all vertebrate species, including, but not
limited to, human, bovine, chicken, mouse, rat, porcine, ovine,
turkey, baboon, and fish (for sequence information, see, e.g.,
McPherron et al. (1997) Proc. Nat. Acad. Sci. U.S.A., 94:
12457-12461).
[0027] The term "mature GDF-8" refers to the protein that is
cleaved from the carboxy-terminal domain of the GDF-8 precursor
protein. The mature GDF-8 may be present as a monomer, homodimer,
or in a GDF-8 latent complex. Depending on conditions, mature GDF-8
may establish equilibrium between any or all of these different
forms. In its biologically active form, the mature GDF-8 is also
referred to as "active GDF-8."
[0028] The term "GDF-8 propeptide" refers to the polypeptide that
is cleaved from the amino-terminal domain of the GDF-8 precursor
protein. The GDF-8 propeptide is capable of binding to the
propeptide binding domain on the mature GDF-8.
[0029] The term "GDF-8 latent complex" refers to the complex of
proteins formed between the mature GDF-8 homodimer and the GDF-8
propeptide. It is believed that two GDF-8 propeptides associate
with two molecules of mature GDF-8 in the homodimer to form an
inactive tetrameric complex. The latent complex may include other
GDF inhibitors in place of or in addition to one or more of the
GDF-8 propeptides.
[0030] The term "GDF-8 activity" refers to one or more of
physiologically growth-regulatory or morphogenetic activities
associated with active GDF-8 protein. For example, active GDF-8 is
a negative regulator of skeletal muscle mass. Active GDF-8 can also
modulate the production of muscle-specific enzymes (e.g., creatine
kinase), stimulate myoblast proliferation, and modulate
preadipocyte differentiation to adipocytes. Exemplary procedures
for measuring GDF-8 activity in vivo and in vitro are found in U.S.
patent application Ser. No. 10/688,925, now U.S. Pat. No.
7,261,893, for example.
[0031] As used herein, "GDF-8 inhibitor" generally refers to any
compound that downregulates the activity of GDF-8, and includes any
agent capable of inhibiting activity, expression, processing, or
secretion of GDF-8. A GDF-8 inhibitor may, for example, affect
stability of or conversion of the precursor molecule to the active,
mature form; interfere with the binding of GDF-8 to one or more
receptors; or interfere with intracellular signaling of the GDF-8
receptor ActRIIB. Such inhibitors include proteins, antibodies,
peptides, peptidomimetics, ribozymes, anti-sense oligonucleotides,
double-stranded RNA, and other small molecules, which specifically
inhibit GDF-8. Such inhibitors are said to "inhibit," "neutralize,"
or "reduce" the biological activity of GDF-8.
[0032] The terms "neutralize," "neutralizing," "inhibitory," and
their cognates refer to a reduction in the activity of GDF-8 by a
GDF-8 inhibitor, relative to the activity of GDF-8 in the absence
of the same inhibitor. The reduction in activity is preferably at
least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or higher.
The methods for assessing neutralizing or inhibitory biological
activity of GDF-8 inhibitors are known in the art, and can be
performed, for example, using the ActRIIB assay (e.g., as described
in Whittemore et al. (2003) BBRC, 300:965-97; or U.S. patent
application Ser. No. 10/253,532, now U.S. Pat. No. 7,320,789) and
the RGA assays (as described in Thies (2001) Growth Factors,
18:251-259 or U.S. patent application Ser. No. 10/253,532, now U.S.
Pat. No. 7,320,789).
[0033] The term "therapeutically effective dose," or
"therapeutically effective amount," refers to that amount of a
compound that results in prevention, reduction in the risk of
occurrence, or amelioration of symptoms in a patient, or a desired
biological outcome, e.g., improved muscle function, delayed onset
of clinical symptoms, etc. The effective amount can be determined
as described in the subsequent sections.
[0034] The terms "treatment," "therapeutic method," and their
cognates refer to treatment or prophylactic/preventative measures.
Those in need of treatment may include individuals already having a
particular medical disorder as well as those who may ultimately
acquire the disorder. Treatment includes any reduction in any
symptom of a disorder described in this application. In addition to
a reduction or lessening of symptoms, treatment also includes
maintaining a patient's current status when worsening is expected,
or preventing the occurrence of a symptom in an individual in which
the onset of a symptom, disorder, or disease is expected. Treatment
may include a decrease or reduction in one or more physiologic
function from normal. It may also include a decrease compared to
expected symptoms or expected progression of the condition,
disorder, or disease.
II. Components for Use in the Methods of the Invention
[0035] In the methods of the present invention, one or more GDF-8
inhibitors are used in combination with one or more
corticosteroids.
[0036] A. GDF-8 Inhibitors
[0037] GDF-8 inhibitors, used in the methods of the present
invention, include, but are not limited to, antibodies to GDF-8;
antibodies to GDF-8 receptors; soluble GDF-8 receptors and
fragments thereof (e.g., ActRIIB fusion polypeptides as described
in U.S. patent application Ser. No. 10/689,677, including soluble
ActRIIB receptors in which ActRIIB is joined to the Fc portion of
an immunoglobulin); GDF-8 propeptide and modified forms thereof
(e.g., as described in WO 02/068650 or U.S. patent application Ser.
No. 10/071,499, now U.S. Pat. No. 7,202,210, including forms in
which GDF-8 propeptide is joined to the Fc portion of an
immunoglobulin and/or forms in which GDF-8 is mutated at an
aspartate (asp) residue, e.g., asp-99 in murine GDF-8 propeptide
and asp-100 in human GDF-8 propeptide); follistatin (e.g., as
described in U.S. Pat. No. 6,004,937) or
follistatin-domain-containing proteins (e.g., GASP-1 or other
proteins as described in U.S. patent application Ser. Nos.
10/369,736 and 10/369,738, now U.S. Pat. Nos. 7,192,717 and
7,572,763); and modulators of metalloprotease activity that affect
GDF-8 activation, as described in U.S. patent application Ser. No.
10/662,438.
[0038] In some embodiments, the GDF-8 inhibitor is a monoclonal
antibody or a fragment thereof that blocks GDF-8 binding to its
receptor. Nonlimiting illustrative embodiments include a nonhuman
monoclonal anti-GDF-8 antibody, e.g., murine monoclonal antibody
JA-16 (as described in U.S. patent application Ser. No. 10/253,532,
now U.S. Pat. No. 7,320,789; ATCC Deposit No. PTA-4236);
derivatives thereof, e.g., humanized antibodies; and fully human
monoclonal anti-GDF-8 antibodies (e.g., Myo29, Myo28, and Myo22, as
described in U.S. patent application Ser. No. 10/688,925, now U.S.
Pat. No. 7,261,893; ATCC Deposit Nos. PTA-4741, PTA-4740, and
PTA-4739, respectively), or derivatives thereof.
[0039] In some embodiments, the GDF-8 inhibitor blocks GDF-8 from
binding to its receptor, by binding to GDF-8 or to the GDF-8
receptor. In various embodiments, the GDF-8 inhibitor is an
anti-GDF-8 antibody that has the affinity to GDF-8, expressed as an
affinity constant (K.sub.a), wherein K.sub.a is at least 10.sup.5
M.sup.-1, 10.sup.6 M.sup.-1, 10.sup.7 M.sup.-1, 10.sup.8 M.sup.-1,
10.sup.9 M.sup.-1, 10.sup.10 M.sup.-1, 10.sup.11 M.sup.-1, or
10.sup.12 M.sup.-1. Also contemplated for use in humans are
inhibitors that are humanized forms and derivatives of nonhuman
antibodies derived from any vertebrate species described in patent
applications cited herein, or in Antibody Engineering, ed.
Borrebaeck, 2nd ed., Oxford University Press, 1995; and Antibodies:
A Laboratory Manual, eds. Harlow et al., Cold Spring Harbor
Laboratory, 1988.
[0040] B. Corticosteroids
[0041] Corticosteroids used in the methods of the present
invention, include, but are not limited to, beclomethasone
dipropionate, budesonide, cortisol, dexamethasone, fluticason
propionate, prednisone, mometasone furoate, triamcinolone
acetonide, and derivatives thereof.
[0042] Pharmaceutically acceptable salts of compounds disclosed
herein can also be used.
[0043] Corticosteroids are available commercially in various
pharmaceutical formulations (Physician's Desk Reference (PDR) 2003,
57th ed., Medical Economics Company, 2002). For example, oral
formulations are commercially available for cortisone,
hydrocortisone (Cortef.RTM.), prednisone (Deltasone.RTM.,
Meticorten.RTM., Orasone.RTM.), prednisolone (Delta-Cortef.RTM.,
Pediapred.RTM., Prelone.RTM.), triamcinolone (Aristocort.RTM.,
Kenacort.RTM.), methylprednisolone (Medrol.RTM.), dexamethasone
(Decadron.RTM., Dexone.RTM., Hexadrol.RTM.), betamethasone
(Celestone.RTM.), and deflazacort (Calcort.RTM.). Other
formulations of these and other corticosteroids can be used in the
methods of the invention.
[0044] C. Therapeutic and Prophylactic Methods
[0045] The invention provides method of treating mammalian
subjects, including methods to treat loss of muscle function,
muscle weakness, and/or corticosteroid-induced muscle atrophy.
[0046] Methods of the invention comprise administering to the
mammal a therapeutically effective amount of at least one GDF-8
inhibitor and a therapeutically effective amount of at least one
corticosteroid in the amounts and for a period of time sufficient
to treat at least one of loss of muscle function, muscle mass,
muscle weakness, muscle atrophy, or cardiomyopathy. The methods can
be used for treating neuromuscular disorders such as muscular
dystrophies. In some embodiments, muscle function is improved
relative to the same treatment either in the absence of the GDF-8
inhibitor or the corticosteroid. The muscles treated include, but
are not limited to, gastrocnemius, tibialis, anterior, quadriceps,
extensor digitorum, cardiac muscle, or diaphragm muscle.
[0047] Neuromuscular disorders include, but are not limited to, any
acute or chronic disease or disorder that compromises muscle
function, causes muscular injury, or otherwise causes a diminution
in muscle mass and/or function. A wide variety of diseases or
disorders is known and includes, for example, muscular dystrophies
such Duchenne muscular dystrophy, Becker muscular dystrophy, Emery
Dreifuss muscular dystrophy, limb girdle muscular dystrophy, rigid
spine syndrome, Ullrich syndrome, Fukuyama muscular dystrophy,
Walker-Warburg syndrome, muscle-eye-brain disease,
facioscapulohumeral muscular dystrophy, congenital muscular
dystrophy, myotonic dystrophy (Steinert's disease), nondystrophic
myotonia, periodic paralyses spinal muscular atrophy, familial
amytrophic lateral sclerosis, hereditary motor and sensory
neuropathy, Charcot-Marie-Tooth disease, chronic inflammatory
neuropathy, distal myopathy, myotubular/centronuclear myopathy,
nemaline myopathy, mini core disease, central core disease,
desminopathy, inclusion body myositis, mitochondrial myopathy,
congenital myasthenic syndrome, post-polio muscle dysfunction, and
disorders described in Emery (2002) The Lancet, 359:687-695; and
Khurana et al. (2003) Nat. Rev. Drug Disc., 2:379-386. Patients may
exhibit mild, moderate or severe muscle weakness, muscle wasting,
and effects on independent ambulation associated with such a
disorder. Patients having or at risk for developing these disorder
will benefit from GDF-8 inhibitor and a corticosteroid.
[0048] In general, a patient who will benefit from coadministration
of a GDF-8 inhibitor and a corticosteroid is one who exhibits a
2-10-fold or higher increase in the serum CK activity, a positive
family history, an abnormal variation in the diameter of muscle
fibers, a deficiency in dystrophin or a mutation in the dystrophin
gene, loss of muscle mass, muscle weakness, cardiomyopathy, and/or
loss of muscle strength. The diagnostic procedures, including the
appropriate genetic testing, are described in Diagnostic Criteria
for Neuromuscular Disorders, ed. Emery, 2nd ed., Royal Society of
Medicine Press, 1997. The combination treatment can be also
beneficial to subjects undergoing corticosteroid therapy for
disorders other than neuromuscular disorders and/or subjects with a
history of a long-term corticosteroid use so long these subjects
exhibit, or are at risk of diminution of muscle function such as
characterized by muscle weakness, loss of muscle mass, and/or
muscle atrophy, etc. Examples of disorders for which corticosteroid
therapy is often used include, but are nor limited to, asthma,
allergy, arthritis, dermatologic disorders (e.g., inflammatory
dermatoses, eczema, psoriasis, etc), lupus erythematosus, and other
chronic inflammatory conditions.
[0049] Methods of administration and compositions used in the
methods of the inventions are provided. Administration is not
limited to any particular delivery system and may include, without
limitation, parenteral (including subcutaneous, intravenous,
intramedullary, intraarticular, intramuscular, or intraperitoneal
injection) rectal, topical, transdermal, or oral (for example, in
capsules, suspensions, or tablets). Administration to an individual
may occur in a single dose or in repeat administrations, and in any
of a variety of physiologically acceptable salt forms, and/or with
an acceptable pharmaceutical carrier and/or additive as part of a
pharmaceutical composition. Physiologically acceptable salt forms
and standard pharmaceutical formulation techniques and excipients
are well known to persons skilled in the art (e.g., as described in
Physician's Desk Reference (PDR) 2003, 57th ed., Medical Economics
Company, 2002; and Remington: The Science and Practice of Pharmacy,
eds. Gennado et al., 20th ed, Lippincott, Williams & Wilkins,
2000).
[0050] A GDF-8 inhibitor and a corticosteroid are administered
concurrently or consecutively over overlapping or nonoverlapping
intervals. In the sequential administration, the GDF-8 inhibitor
and the corticosteroid can be administered in any order. In some
embodiments, the length of an overlapping or nonoverlapping
interval is more than 2, 4, 6, 12, 24, or 48 weeks.
[0051] For corticosteroids, the prescribing physician routinely
selects the dosage and regimen. For example, prednisone is used at
about 0.1-2 mg per kilogram of body weight per day, and most
commonly at 0.5-1 mg/kg/day, e.g., 0.75 mg/kg/day. The
corticosteroid may be administered at average weekly doses of
approximately 1-14 mg/kg body weight, including approximately 1, 2,
5, 7, 10, 12, or 15 mg/kg body weight per week, and the prescribing
physician may select a frequency of administration as appropriate.
Single dose, continuous or periodic corticosteroid administration
may be selected, including administration at hourly, daily,
bi-weekly, weekly, or other periodic intervals. Preferably,
corticosteroids are administered orally or by injection 1-4 times
per day. Corticosteroid dosage may be optimized as a combination
therapy, and dosage may be lowered to reduce significant side
effects of administration.
[0052] The GDF-8 inhibitors can be administered alone or in a
mixture with a corticosteroid or another compound. GDF-8 inhibitors
can be administered at a dose of approximately from 1 .mu.g/kg to
25 mg/kg, depending on physiology, the severity of the symptoms and
the progression of the disease. Single dose, continuous, or
periodic administration may be selected, with intervals between
GDF-8 inhibitor doses chosen from hourly, daily, bi-weekly, weekly,
bi-monthly, monthly, or other appropriate intervals. For example,
GDF-8 inhibitors such as antibodies may be administered in an
outpatient setting by weekly administration at about 0.1-10 mg/kg
dose by intravenous (IV) infusion, intraperitoneal, or subcutaneous
injection. In general, the appropriate therapeutically effective
dose of a GDF-8 inhibitor is selected by a treating clinician and
would range approximately from 1 .mu.g/kg to 20 mg/kg, from 1
.mu.g/kg to 10 mg/kg, from 1 .mu.g/kg to 1 mg/kg, from 10 .mu.g/kg
to 1 mg/kg, from 10 .mu.g/kg to 100 .mu.g/kg, from 100 .mu.g to 1
mg/kg, and from 500 .mu.g/kg to 5 mg/kg. Exemplary effective doses
of GDF-8 inhibitor include approximately 0.1, 0.3, 0.5, 1, 5, 10,
or 20 mg/kg/wk. Additionally, specific dosages indicated in the
Examples or in the Physician's Desk Reference (PDR) 2003, 57th ed.,
Medical Economics Company, 2002, can be used.
[0053] D. Methods of Testing Compounds for Therapeutic Efficacy
[0054] The invention further provides methods for testing in an
animal, e.g., a rodent or a primate, whether a therapeutic compound
is efficacious when administered in combination with at least one
GDF-8 inhibitor and at least one corticosteroid. In some
embodiments, the method of evaluating the efficacy of a compound
comprises: administering the compound to a first animal in
combination with a GDF-8 inhibitor and a corticosteroid;
administering the GDF-8 inhibitor and the corticosteroid to a
second animal; determining the level of muscle function in the
first and in the second animal after the administrations; and
comparing the levels of muscle function. If the level in the first
animal is lower than the level in the second animal, it indicates
that the compound or the combination is efficacious.
[0055] In other embodiments, the compound may be evaluated for
efficacy in treatment of muscular dystrophy when administered in
combination with a GDF-8 inhibitor and/or a corticosteroid.
[0056] Several animal models are available for such evaluative
purposes. For example, the mdx model has been described, for
example, by Torres et al. (1987) Brain, 110:269-299, and Hoffman et
al. (1987) Science, 238:347-350. Extremely high levels of CK are
consistently noted with dystrophin-deficiency in mdx mice and DMD
humans due to sarcolemmal damage (Bulfield et al. (1984) Proc.
Natl. Acad. Sci. USA, 81:1189-1192; and Matsuda et al. (1995) J.
Biochem. (Tokyo), 118: 959-64). As another example, two other
animal models can be used: utr-/- mdx mice (Gillis (2002)
Neuromuscul. Disord., 12(1):90-84; and Deconick et al. (1997) Cell,
90:729-738) and nu-/- mdx mice (Morrison et al. (2000) Lab.
Invest., 80:881-891).
EXAMPLES
Example 1
Effect of GDF-8 Neutralizing Antibody on Dystrophic Muscle
[0057] The ability of in vivo inhibition of GDF-8 to ameliorate
muscular dystrophy was tested in the mdx mouse model of DMD. Five
to seven week old male C57BL/10ScSn-mdx/j mice (Jackson Laboratory,
Bar Harbor, Me.) were treated with weekly intraperitoneal
injections of the GDF-8 neutralizing murine antibody JA-16 (60
mg/kg, double dosing at first week, n=11), and vehicle alone
(control group, n=10) for 12 weeks. These mice were also compared
to mice of the same background strain (C57BL/10, n=12) without the
dystrophin deficiency.
[0058] The body weight was monitored before, during and after
treatment. Mice in the treatment group gained weight relative to
mice in the vehicle control group. Results are shown in Table
1.
TABLE-US-00001 TABLE 1 Total body weight (g) Average values with
SEM Week JA-16 vehicle control vehicle control of trial (mdx) (mdx)
(non-mdx) 0 21.92 +/- 0.42 22.51 +/- 0.36 19.18 +/- 0.40 4 27.82
+/- 0.43 26.76 +/- 0.60 24.14 +/- 0.27 8 29.59 +/- 0.54 28.49 +/-
0.58 25.31 +/- 0.28 12 32.42 +/- 0.57 31.12 +/- 0.73 27.17 +/-
0.39
[0059] Mice were also subjected to a grip test after 6 and 10 weeks
of dosing. Mice in the treatment group at four and ten weeks had 9%
(p=0.09) and 19% (p<0.05) respectively greater grip strength
than mice in the vehicle control groups. Results are shown in Table
2.
TABLE-US-00002 TABLE 2 Grip strength (lb) Average values with SEM
Week JA-16 vehicle control vehicle control of trial (mdx) (mdx)
(non-mdx) 6 0.261 +/- 0.011 0.239 +/- 0.006 0.239 +/- 0.011 10
0.249 +/- 0.006 0.210 +/- 0.014 0.247 +/- 0.010
[0060] To quantify the difference in muscle mass between treatment
and vehicle control, animals were sacrificed and quadriceps and
gastrocnemius muscles dissected out and weighed. Quadriceps muscles
from the treated group of animals weighed 13% more than controls
(0.371.+-.0.009 vs. 0.317.+-.0.008 g; p<0.05). Gastrocnemius
muscles from the treated group of animals weighed 17% more than
controls (0.223.+-.0.008 vs. 0.197.+-.0.005 g; p<0.0005).
Example 2
Effect of GDF-8 Neutralizing Antibody and Prednisone on Normal and
Dystrophic Muscle
[0061] Male C57BL/10ScSn-mdx/j and C57BL/10 (Jackson Laboratory,
Bar Harbor, Me.). Mouse monoclonal anti-GDF-8 antibody JA-16,
prednisone (P-9901, Sigma), or vehicle (peanut oil) was injected
starting at age 5-7 weeks for 4 weeks. Mice were intraperitoneally
(IP) injected with JA-16 at a dose of 60 mg/kg per week (double
dosing at first week), or subcutaneously (SC) injected with
prednisone at 2 mg/kg, 3 times a week.
[0062] The body weight and grip strength were monitored before,
during and after treatment. Results are shown in Table 3 and Table
4, respectively.
TABLE-US-00003 TABLE 3 Total body weight (average .+-. SEM, g) Week
Prednisone + vehicle vehicle of JA-16 Prednisone control control
trial (mdx) (mdx) (mdx) (non-mdx) 0 17.7 .+-. 1.6 17.6 .+-. 1.8
16.0 .+-. 1.9 19.2 .+-. 0.4 1 22.1 .+-. 1.4 20.9 .+-. 1.8 19.1 .+-.
2.1 22.4 .+-. 0.3 2 25.9 .+-. 1.2 24.2 .+-. 1.6 23.7 .+-. 1.4 23.8
.+-. 0.4 3 26.5 .+-. 1.1 24.7 .+-. 1.5 24.8 .+-. 1.2 24.9 .+-. 0.4
4 28.1 .+-. 1.2 25.9 .+-. 1.6 25.7 .+-. 1.4 25.6 .+-. 0.5
TABLE-US-00004 TABLE 4 Grip strength (average .+-. SEM lb) Week
Prednisone + vehicle vehicle of JA-16 Prednisone control control
trial (mdx) (mdx) (mdx) (non-mdx) 0 0.161 .+-. 0.018 0.144 .+-.
0.010 0.164 .+-. 0.014 0.164 .+-. 0.009 3 0.219 .+-. 0.019 0.177
.+-. 0.006 0.168 .+-. 0.005 0.212 .+-. 0.010 4 0.281 .+-. 0.011
0.213 .+-. 0.011 0.217 .+-. 0.010 0.234 .+-. 0.018
[0063] At the end of the study, mice were sacrificed and muscle
mass was assessed by dissecting and weighing the gastrocnemius and
quadriceps. Results are shown in Table 5. To confirm biological
activity of prednisone, sera from a separate cohort of mice were
collected and analyzed for IL6 and IL1.beta. (Ani Lytics, Inc.,
Gaithersburg, Md.). Both cytokines were found to be reduced in the
sera of mice treated with prednisone.
TABLE-US-00005 TABLE 5 Muscle weight (average .+-. SEM, g)
Prednisone + vehicle vehicle Mus- JA-16 Prednisone control control
cle (mdx) (mdx) (mdx) (non-mdx) Gas- 0.364 .+-. 0.019 0.287 .+-.
0.023 0.299 .+-. 0.019 0.280 .+-. 0.010 troc Quad 0.527 .+-. 0.030
0.417 .+-. 0.029 0.415 .+-. 0.030 0.392 .+-. 0.010
[0064] Therefore, the results demonstrate that in muscular
dystrophy, administration of an inhibitor of GDF-8, i.e.,
anti-GDF-8 antibody, and prednisone is effective in increasing
muscle mass and strength relative to treatment with prednisone
alone or vehicle.
[0065] Furthermore, in these studies the effects of JA16 plus
prednisone treatment (Example 2) were greater than the effects of
treatment with JA16 alone (Example 1). The increase in body weight
compared to vehicle after four weeks of treatment was more dramatic
for JA16 plus prednisone treatment than for JA16 treatment alone.
The increase in grip strength compared to vehicle control after
four weeks of treatment with JA16 plus prednisone was greater that
the increase after six or ten weeks of treatment with JA16 alone.
The increase over vehicle control in muscle mass after four weeks
of treatment with JA16 plus prednisone was also greater than the
increase after twelve weeks of treatment with JA16 alone.
Example 3
Effect of GDF-8 Neutralizing Antibody on Prednisone-Induced Muscle
Atrophy
[0066] In the mice treated as described in Example 2, diaphragm
muscle was histologically examined as described in Example 1. The
morphological changes were evaluated by an independent pathology
lab that had no knowledge of the treatment group assignments.
Severity grades were assigned on a scale from 0 to 4 (0=none;
1=minimal; 2=mild; 3=moderate; and 4=marked). Results are shown in
FIG. 1A (severity scores) and FIG. 1B (percentage of muscle fibers
atrophied). The results show that administration of the anti-GDF-8
antibody with prednisone reduces prednisone-induced muscle
atrophy.
Example 4
Treatment of Muscular Dystrophies
[0067] As an example of treating MD in humans, the Myo29 antibody
is administered in combination with prednisone or prednisolone.
Nonlimiting exemplary treatment regimens and outcomes are
summarized in Table 6. Other treatment regimens can be determined
by a treating physician, with ranges of the corticosteroids and
GDF-8 inhibitors dosage and administration as discussed above.
TABLE-US-00006 TABLE 6 Patient No. Treatment Regimen Treatment Goal
Patient 1 Myo29 at 10 mg/kg/week, Maintenance and/or increase
administered by bi-weekly of muscle mass, strength, and injection
plus prednisone function over benefit of at 0.75 mg/kg/day for 2
prednisone alone years, or continuing treatment as needed Patient 2
Myo29 at 0.1 mg/kg/week, Maintenance and/or increase administered
by weekly IV of muscle mass, strength, and plus prednisone at 1.0
function over benefit of mg/kg/day, continuing prednisone alone
treatment as needed. Patient 3 Myo29 at 1 mg/kg/week Maintenance
and/or increase administered by monthly of muscle mass, strength
and injection plus prednisone function or increased at 0.50
mg/kg/day for 2 preservation of function for years, or continuing
muscle groups that are not treatment as needed already compromised
over benefit of prednisone alone Patient 4 Myo29 at 20 mg/kg/week,
Maintenance and/or increase administered in a single of muscle
mass, strength and dose by IV plus prednisone function or increased
at 0.75 mg/kg/day for 2 preservation of function over years, or
continuing benefit of prednisone alone treatment as needed Patient
5 Myo29 at .1 mg/kg/week, Maintenance and/or increase administered
in a single of muscle mass, strength and dose by IV plus prednisone
function over benefit of at 5 mg/kg/wk, as needed prednisone alone
Patient 6 Myo29 at 1 mg/kg/week, Maintenance and/or increase
administered weekly by IV of muscle mass, strength and plus
prednisone at 2 function over benefit of mg/kg/wk for at least 2
prednisone alone months, or as needed Patient 7 Myo29 at 10
mg/kg/week, Maintenance and/or increase administered in a single of
muscle mass, strength and dose by subcutaneous function over
benefit of injection plus prednisone prednisone alone at 7 mg/kg/wk
for at least 6 months, or continuing treatment as needed Patient 8
Myo29 at 20 mg/kg/week, Maintenance and/or increase administered
weekly by of muscle mass, strength and injection plus prednisone
function over benefit of at 14 mg/kg/wk for 2 years, prednisone
alone or continuing treatment as needed Patient 9 Myo29 at 1
mg/kg/week, Maintenance and/or increase administered bi-monthly by
of muscle mass, strength and IV plus prednisone at 10 function over
benefit of mg/kg/wk for at least 1 prednisone alone month, or
continuing treatment as needed Patient 10 Myo29 at 0.3 mg/kg/week,
Maintenance and/or increase administered monthly by of muscle mass,
strength and subcutaneous injection function or increased plus
prednisone at 0.75 preservation of function for mg/kg/day for 1
year, or muscle groups over benefit of continuing treatment as
prednisone alone needed
[0068] All publications and patents cited and sequences identified
by accession or database reference numbers in this disclosure are
incorporated by reference in their entirety.
* * * * *