WO 2006/105359 PCT/US2006/011799
METHODS FOR STIMULATING HAIR GROWTH BY ADMINISTERING BMPs
DESCRIPTION OF THE INVENTION
FIELD OF THE INVENTION
[001] The present invention relates to methods of stimulating hair growth, .
and to pharmaceutical compositions that stimulate hair growth. The present
invention also relates to methods of inhibiting the immune system, and to
pharmaceutical compositions that the immune system.
BACKGROUND OF THE INVENTION
[002] Tens of millions of Americans suffer from some type of hair loss. A
wide variety of conditions cause hair loss, including androgenic alopecia, or
common pattern baldness, anagen effluvium a chemotherapy-induced hair loss,
telogen effluvium, induced by stress, fever and drugs and alopecia areata, an .
autoimmune disease which afflicts an estimated four million people. Cotsarelis et
al., "Towards a molecular understanding of hair loss and its treatment," TRENDS
in Mol. Med. 7(7):293-301 (2001); NATIONAL INSTITUTES OF HEALTH, Questions &
Answers About Alopecia Areata (2003); Mac Donald, N., "Alopecia areata:
identification and current treatment approaches," Dermatol. Nurs. 11:356-359
(1999). While not life-threatening, hair-related disorders affect personal
appearance, and therefore frequently have profound impact on patients' social
interactions, self-esteem, and psychological well being.
[003] Androgenic alopecia affects both men and women, although women
tend to lose less hair, and in a more diffuse pattern than men. There is also
evidence that androgenic hormones, such as testosterone, coupled with a genetic
predisposition, are necessary for the development of male pattern baldness. It is
currently believed that the conversion of testosterone into dihydrotestosterone, a
compound which inhibits hair growth, by the enzyme 5-a-reductase, triggers
pattern baldness in men, but the mechanism of interaction'between the hormone
and hair follicles remains unknown/ Female pattern baldness is thought to result
from a decrease in estrogen, a hormone that normally counteracts the balding
effect of testosterone, although there is so far no consensus on whether pattern
baldness in women is truly androgen-dependent. Vierhapper et al., "Production
rates of testosterone and of dihydrotestosterone in female pattern hair loss,"
Metabolism 52(7):927-929 (2003).
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[004] Telogen effluvium manifests as excessive shedding of hair, which
occurs as cycling hair follicles prematurely enter the resting phase of the hair
growth cycle, called telogen. It may be precipitated by a multitude of stress-
related causes, including high fevers, childbirth, severe infections, severe chronic
illness, severe psychological stress, major surgery, an over- or under-active
thyroid gland, crash diets with inadequate protein, and a variety of medications,
including, e.g., retinoids, beta blockers, calcium channel blockers,
antidepressants, and non-steroidal anti-inflammatories, including ibuprofen and
acetominophen. Generally little treatment is possible beyond identifying and
either treating or discontinuing the inciting factor, whichever is appropriate. In
most cases, the lost hair will be replaced within six to twelve months. Paus, et al.,
"The biology of hair follicles," N. Engl. J. Med. 341(7):491-497 (1999).
[005] Most cases of drug-induced alopecia involve normal hairs entering
telogen prematurely, as in telogen effluvium. In contrast, anagen effluvium, the
most common type of chemotherapy-induced alopecia, results from the abrupt
cessation of mitotic activity in hair matrix cells of anagen hair follicles. This
induces the follicles to produce either no hair, or produce only narrow defective
hair sheaths which are predisposed to fracture and loss. This type of alopecia can
be seen to some degree in most anti-neoplastic therapies, depending on dosage
and route of administration. However, there are certain agents, such as
bleomycin, cisplatin, doxorubicin, vinblastine and vincristine, which induce
alopecia more frequently and severely. These agents display a synergistic effect
when used in combination and may cause severe and complete alopecia.
Anagen effluvium manifests within 1 to 2 weeks after the beginning of
chemotherapy but is most noticeable 1 to 2 months later. Initially, there may not
be total hair loss, since approximately 10% of follicles will not be in anagen phase
at the start of chemotherapy. Total hair loss eventually occurs with prolonged
therapy, which can also induce hair loss in other areas of the body. Hair regrowth
can usually be expected after the end of chemotherapy, although hair color and
texture may change.
[006] Alopecia areata usually presents as varying amounts of patchy hair
loss, most commonly on the scalp (though it can affect any hair-bearing surface),
but may also manifest as larger patches with little or no hair. Related forms of the
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disease include: (1) alopecia totalis, characterized by complete loss of all scalp
hair; and (2) alopecia universalis, characterized by loss of all body hair, including
eyelashes, eyebrows, underarm hair, and pubic hair. The latter form can cause
serious respiratory problems because the nostrils and sinuses are no longer
protected from airborne foreign particles. Hull et alM "Guidelines for the
management of alopecia areata," Brit. J. Dermatol. 149:692-699 (2003).
[007] Alopecia areata is an autoimmune disease in which cells of the
anagen hair bulb are attacked by T lymphocytes. In a process resembling
chemotherapy-induced hair loss, lymphocyte infiltration of the growing hair bulb
forces the anagen follicles into dystrophic catagen, causing the hair shaft to break
off. Possible targets of autoimmune attack in alopecia areata include matrix
keratinocytes, dermal papilla cells, and melanocytes. Cotsarelis et al., supra.
Linkage analyses indicate that this disease has a genetic component, though the
range of associated genes, including the major histocompatibility complex,
cytokine and immunoglobulin genes, suggests that any genetic predisposition is
likely multifactorial. Hull et al., supra. In any case, whether the underlying defect
in alopecia areata lies within the hair follicle, the immune system, or both, is not
known. Kalish, et al., "Alopecia, areata: autoimmunity—the evidence is.
compelling," J. Invest. Dermatol. 8(2):164-167 (2003).
[008] Medications approved for other purposes can help hair grow back in
those suffering from alopecia areata, at least temporarily, though none cure the
underlying disease. NATIONAL INSTITUTES OF HEALTH, supra. For example,
patients may be treated with corticosteroids {e.g., prednisone, dexamethasone, or
hydrocortisone) administered orally, topically, or by injection, with oral finasteride,
or with a topical solution of minbxidil. Because alopecia areata is an autoimmune
disorder, patients are sometimes treated with immunosuppressive compounds as
well, (see, e.g., U.S. Patent No. 5,342,625; U.S. Patent No. 5,284,826; and U.S.
Patent No. 4,996,193, describing the use of cyclosporin A and related
immunosuppressive compounds for hair revitalization, and citing the known use of
cyclosporin and related immunosuppressive compounds for hair growth), although
such drugs often have significant toxic side effects.
[009] There are two drugs currently approved by the Food & Drug
Administration (FDA) for the treatment of male pattern baldness: Rogaine®
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(topical minoxidil) and Propecia® (oral finasteride). Both were initially used to treat
other medical conditions. Minoxidil, a potassium channel agonist that potently
induces peripheral vasodilation, was originally used as a treatment for
hypertension. The mechanism by which minoxidil induces hair growth is
unknown. Dormois et al., "Minoxidil in severe hypertension: value when
conventional drugs have failed," Am. Heart J. 90:360-368 (1975); Messenger,
A.G. et al., "Minoxidil: mechanisms of action on hair growth," Brit. J. Dermatol.
150:186-194 (2004). Finasteride was originally used to treat urinary problems
. caused by enlargement of the prostate in men (called benign prostatic
hyperplasia). It blocks the activity of 5-a-reductase, an enzyme that converts
testosterone to dihydrotestosterone (DHT), a more active form of the hormone
which has been implicated in miniaturization of hairs, a precursor to catagen.
Brown et al., "A current review of medical therapy for benign prostatic
hyperplasia," J.Am. Osteopath, Assoc, 104(S2):S11-S16(2004).
[010] Minoxidil and finasteride both stimulate hair regrowth in some
patients, but only for the duration of drug use: new hair growth ends arid hair loss
resumes shortly after the patient stops treatment. After several months' use,
minoxidil successfully induces limited hair growth for approximately 1 in 3 patients,
and slows hair loss for roughly 9 in 10. Physician's Desk Reference® 2580 (49th
ed. 1995). Oral finasteride is generally more effective than topical minoxidil at
inducing hair growth, but both treatments are far less than 100% effective.
Further hair loss is prevented in most patients treated with finasteride. About half
of treated patients achieve some hair regrowth, and approximately one-third of
patients experience cosmetically important hair regrowth after two years of
continuous use. Foley, P.A., "Recent advances: dermatology," Brit. Med. J.
320:850-853 (2000).
[011] Both minoxidil and finasteride are sometimes accompanied by a .
number of potentially serious side effects. Possible side effects of minoxidil
include: scalp itching or rash; headaches; dizziness; decreased libido; elevated
heart rate; difficulty breathing; and weight gain. Physician's Desk Reference®
2581 (49th ed. 1995). Possible side effects of finasteride include: skin rash;
breast enlargement or tenderness; swelling of lips; testicular pain; decreased
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libido; decreased volume of ejaculate; and impotence. Physician's Desk
Reference® 2067-2069 (58th ed. 2004).
[012] A number of newer methods of treating hair loss employ topical
formulations of a variety of compounds, including nucleic acids and various small
molecules: (1) a nucleotide sequence encoding the cyclin-dependent kinase
inhibitor p21 (U.S. Patent No. 6,844,326); (2) estrogen receptor antagonists (U.S.
Patent Nos. 6,555,532; 6,204,258; and 5,965,551); (3) a modified, no.n-
immunosuppressive form of Cyclosporin A (U.S. Patent No. 6,521,595); (4)
ketoconazole (U.S. Patent No. 6,482,826); and (5) compositions containing an
aliphatic, alkoxy- or aryl-substituted cyclopropenone (U.S. Patent No. 4,985,464).
None of these treatments has yet received FDA approval.
[013] Some patients seek therapeutic options in addition to drug
treatment, including surgical intervention. The most common surgical treatment
for hair loss is transplantation, which transfers grafts of skin and hair from the
back of the scalp, where hair growth is full, to bald areas. Transplantation may
employ mini- or micro-grafts. By this technique,, as few as one or two hairs are
transplanted with each graft, with 100 or more grafts performed per session. This
technique provides a more natural hairline but requires more grafts, and hence
more time, than other methods. Bernstein, et al., "The aesthetics of follicular
transplantation," Dermatol. Surg. 23(9):785-799 (1997). A less common surgical
treatment for hair loss is scalp reduction/which involves removing areas of bald
scalp to bring existing areas of hair growth closer together. Sometimes the skin of
the scalp is too tight for this, and alternative treatments must be used.
[014] Autoimmune diseases result from abnormalities in immune cell
function or activity which cause inappropriately activated T cells to react against
self tissue, thereby triggering production of cytokines or autoantibpdies
responsible, for disease etiology and progression. Autoimmune disorders may be.
systemic, affecting multiple organs or tissues, or localized, affecting a single
organ, organ system or tissue. Limited treatment options focus on: (1) relieving
symptoms, whether by administration of analgesics or non-steroidal anti-
inflammatory drugs or by surgery; (2) preserving organ function, for example by
treating a patient suffering from diabetes mellitus with insulin injections; or (3)
targeting disease mechanisms by suppressing the immune system. These
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treatment options are generally unsatisfactory, because none of them cure the
underlying disease, but only ameliorate the symptoms temporarily. In addition,
prolonged use of immunosuppressive drugs frequently results in secondary
infections, because patients' immune systems cannot repel commonly
encountered fungal, bacterial, or viral pathogens.
SUMMARY OF THE INVENTION
[015] The present invention provides methods for treating hair loss.
disorders by administration of BMP compositions. The invention further provides
methods of treating hair loss disorders by administration of BMP compositions in
combination with other compounds used to treat such disorders, including
corticosteroids, calcineurin inhibitors, topical minoxidil, and oral finasteride. Some
aspects of the invention provide different pharmaceutical formulations of BMP
compositions to facilitate different routes of administration. Other aspects of the
invention provide different dosage ranges or treatment regimens to treat a wide
range of hair loss disorders. Exemplary hair loss disorders that may be treated
with the compositions of the invention include alopecia areata, alopecia totalis,
alopecia universalis, androgenic alopecia, telogen effluvium, anagen effluvium,
and chemotherapy-induced alopecia.
[016] Methods for treating autoimmune disorders by administration of
BMP compositions are also provided. The invention further provides methods of
treating autoimmune disorders by administration of BMP compositions in
combination with other compounds used to treat such disorders, including
calcineurin inhibitors and other compounds with known immunosuppressive
activity. Some aspects of the invention provide different pharmaceutical
formulations of BMP compositions to facilitate different routes of administration.
Other aspects of the invention provide different dosage ranges or treatment
regimens to treat a wide range of autoimmune disorders. Exemplary autoimmune
disorders that may be treated with the compositions of the invention include
Crohn's disease, inflammatory bowel disease, multiple sclerosis, psoriasis,
rheumatoid arthritis, or systemic lupus erythematosus.
[017] Embodiments of the invention include, without limitation, the
following.
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[018] Use of a composition comprising a therapeutically effective amount
of at least one BMP chosen from the group consisting of BMP-2, BMP-3, BMP-4,
BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13,
BMP-14, BMP-15, BMP-16, BMP-17, BMP-18,:and MP52/GDF-5 for manufacture
of a medicament for promoting hair growth in a patient suffering from a hair loss
disorder.
[019] Use of a therapeutically effective amount of at least one BMP.
chosen from the group consisting of BMP-2, BMP-3, BMP-4, BMP-5, BMP-6;
BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13, BMP-14, BMP-15,
BMP-16, BMP-17, BMP-18, and MP52/GDF-5; and a therapeutically effective
amount of at least one compound chosen from the group comprising: prednisone,
dexamethisone, hydrocortisone, cyclosporin A, pimecrolimus, tacrolimus,
minoxidil, and finasteride;for manufacture of a medicament for promoting hair
growth in a patient suffering from a hair loss disorder.
[020] Use of a composition comprising a therapeutically effective amount
of at least one BMP chosen from the group consisting of BMP-2, BMP-3, BMP-4,
BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13,
BMP-14, BMP-15, BMP-16, BMP-17, BMP-18, and MP52/GDF-5 for manufacture
of a medicament for promoting hair growth.
[021] Use of a composition comprising therapeutically effective amount
of at least one BMP chosen from the group consisting of BMP-2, BMP-3, BMP-4,
BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13,
BMP-14, BMP-15, BMP-16, BMP-17, BMP-18, and MP52/GDF-5 for cosmetic
treatment.
[022] Pharmaceutical compositions comprising at least one BMP chosen
from the group consisting of BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7,
BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13, BMP-14, BMP-15, BMP-16,
BMP-17, BMP-18, and MP52/GDF^5 and a sterile solution of 0.5% sucrose, 2.5%
glycirie, 5 mM L-glutamic acid, 5 mM NaCI, and 0.01% polysorbate 80, at pH 4.50.
[023] Pharmaceutical compositions comprising at least one BMP chosen
from the group consisting of BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7,
BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13, BMP-14, BMP-15, BMP-16,
BMP-17, BMP-18, and MP52/GDF-5; and at least one compound chosen from the
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group comprising: prednisone, dexamethisone, hydrocortisone, cyclosporin A,
pimec.rolimus, tacrolimus, minoxidil, and finasteride.
[024] The hair loss disorder may include, e.g., alopecia areata, alopecia
toialis, alopecia universalis, androgenic alopecia, telogen effluvium, ar.ageh
effluvium, and chemotherapy-induced alopecia.
[025] Use of a composition comprising a therapeutically effective amount
of at least one BMP chosen from the group consisting of BMP-2," BMP-3, BMP-4,
BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13,
BMP-14, BMP-15, BMP-16, BMP-17, BMP-18, and MP52/GDF-5 for manufacture
of a medicament for inhibiting immune system activity in a patient suffering from
an autoimmune disease.
[026] Use of a composition comprising a therapeuticaiiy effective amount
of at least one BMP chosen from the group consisting of BMP-2, BMP-3, BMP-4,
BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13,
BMP-14, BMP-15, BMP-16, BMP-17, BMP-18, and MP52/GDF-5; and a
therapeutically effective amount of at least one compound chosen from the group
consisting of cyclosporin A, pimecrolimus, tacrolimus, azathioprine,
mycophenolate mofetil, rapamycin, CCI-779, methotrexate, leflunomide,
interferon-B, copaxone, budenoside, epidermal growth factor, sulfasalazine, 6-
mercaptopurine, azathioprine, metronidazole, mesalamine, olsalazine,
ciprofloxacin, and lignocaine; for manufacture of a medicament for inhibiting
immune system activity in a patient suffering from an autoimmune disorder.
[027] Pharmaceutical compositions comprising.at least one BMP chosen
from the group consisting of BMP-2, BMP-3, BMP-4, BMP:5, BMP-6, BMP-7,
BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13, BMP-14, BMP-15, BMP-16,
BMP-17, BMP-18, and MP52/GDF-5; and at least one compound chosen from the
group consisting of cyclosporin A, pimecrolimus, tacrolimus, azathioprine,
mycophenolate mofetil, rapamycin, CCI-779, methotrexate, leflunomide,
interferon-8, copaxone, budenoside, epidermal growth factor, sulfasalazine, 6-
mercaptopurine, azathioprine, metronidazole, mesalamine, olsalazine,
ciprofloxacin, and lignocaine.
[028] The autoimmune disorder may be chosen from the group
consisting of ankylosing spondylitis, antiphospholipid syndrome, Addison's
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Disease, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune
lymphoproliferative syndrome (ALPS), autoimmune thrombocytopenic purpura
(ATP), Behcet's Disease, bulbous pemphigoid, cardiomyopathy, celiac disease,
Dermatitis Herpetiformis, chronic fatigue immune dysfunction syndrome (CFiDS),
chronic inflammatory demyelinating polyneuropathy (CIDP), cicatricial
pemphigoid, systemic sclerosis (CREST Syndrome), cold agglutinin disease,
Crohn's Disease, cutaneous vasculitis, Degos1 Disease, dermatomyositis, juvenile
dermatomyositis, discoid lupus erythematosus, essential mixed cryoglobulinemia,
fibromyalgia, Goodpasture's Syndrome, Graves' Disease, Guillain-Barre
Syndrome, Hashimoto's Thyroiditis, idiopathic pulmonary fibrosis, idiopathic
thrombocytopenia purpura (ITP), immunoglobulin A nephropathy, insulin-
dependent diabetes mellitus, juvenile arthritis, Kawasaki's Disease, lichen planus,
membranous glomerulonephritis, Meniere's Disease, mixed connective tissue
disease, multifocal motor neuropathy, multiple sclerosis, myasthenia gravis,
pemphigus vuigaris, pernicious anemia, polyarteritis nodosa, polychondritis,
pdlyglandular syndrome, polymyalgia rheumatica, pplymyositis and
dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis,
psoriasis, Raynaud's Phenomenon, Reiter's Syndrome, rheumatic fever,
rheumatoid arthritis, sarcoidosis, scleroderma, Sjogren's Syndrome, Stiff-Man
Syndrome, systemic lupus erthryomatosus, Takayasu arteritis, temporal
arteritis/giant cell arteritis, ulcerative colitis, uveitis, vasculitis, vitiligo, and
Wegener*s Granulomatosis.
[029] The composition may comprise a therapeutically effective amounts
of both BMP-2 and BMP-4. The therapeutically effective amount of a BMP may
be chosen from about 0.05 to about 500 mg, from about 0.5 to about 50 mg, from
about 1 to about 25 mg, and from about 5 to about 10 mg. The therapeutically
effective amount of the BMP may be administered at a concentration chosen from
about 0.001 mg/ml to about 100 mg/ml, from about 0.01 mg/ml to about 50 mg/ml,
from about 0.1 mg/ml to about 25 mg/ml, and from about 1 mg/ml to about 5
mg/ml. The medicament may beadministered to the patient at intervals chosen
from: daily, weekly, monthly,.bi-monthly, tri-monthly, bi-annually, and annually, for
a period of time chosen from: about 1 to about 4 weeks, about 5 to about 24
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weeks, about 25 to about 52 weeks, about 1 to about 2 years, about 2 to about 5
years, about 5 to about 10 years and about 10 to about 20 years.
[030] The medicament may be administered by injection, such as, e.g.,
intraqsseousiy, intravenously, parenterally, percutaneously or extra-corporea'.iy.
[031] The patient may be a mammal, such as, e.g., a human.
[032] Additional objects and advantages of the invention will be set forth in
part in the description which follows, and in part will be obvious from the .
description, or may be learned by practice of the invention. The objects and
advantages of the invention will be realized and attained by means of the
elements and combinations particularly pointed out in the appended claims.
[033] 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
[034] Figure 1 shows a photo of patient 602/J-T after treatment with
recombinant human BMP-2.
DETAILED DESCRIPTION OF THE INVENTION
[035] 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.
[036] As used herein, the term "autoimmune disorder" includes
disorders and diseases caused by abnormalities in any immune cell or immune
cell function or activity, or any disease or disorder characterized by aberrantly or
abnormally elevated immune response. Autoimmune disorders may be systemic,
affecting multiple organs or tissues, or localized, affecting a single organ or tissue.
Examples of systemic autoimmune diseases include, but are not limited to,
rheumatoid arthritis, systemic lupus erythematosus, scleroderma, Sjogren's
syndrome, Goodpasture's syndrome, Wegener's granulbmatosis, polymyalgia
rheumatica, and temporal arteritis or giant cell arteritis. Examples of localized
autoimmune diseases include, but are not limited to, alopecia areata, type 1
diabetes mellitus, Hashimoto's thyroiditis, Graves' disease, ulcerative colitis,
Crohn's disease, celiac disease, multiple sclerosis, Guillain-Barre syndrome,
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Addison's disease, primary biliary sclerosis, sclerosing cholangitis, autoimmune
hepatitis, and Raynaud's phenomenon.
[037] As used herein, the term "BMP activity" refers to biological activity
with the ability to stimulate hair growth or to inhibit immune system activity
performed by a BMP composition in a biological system, which may refer to in
vitro or in vivo models or therapeutic subjects, depending on the context.
[038] As used herein, the terms "bone morphogenetic protein" or
"BMP" refer to any mammalian gene, RNA, or protein of the BMP family of TGF-β
proteins, including, but not limited to, BMPs 2-18 and MP52/GDF-5. In particular,
a BMP will have an identifying pattern of seven conserved cysteine residues in the
mature, carboxy-terminal portion of the protein, as described in Rosen et al.,
"Bone Morphogenetic Proteins" Principles of Bone Biology 2:919-928 (2002); and
Wozney, J.M., "Bone morphogenetic proteins and their gene expression,"
CELLULAR AND MOLECULAR BIOLOGY OF BONE 131-167 (Noda, M. ed. 1993). These
terms also refer to variants, allelic variants, fragments of, and mutant BMPs,
including but not limited to deletion mutants, insertion mutants, and substitution
mutants sharing at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%. and.
99% amino acid sequence identity with a full-length BMP, or having conservative
substitutions at 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, and 1% of amino acid
residues, excluding the seven conserved cysteine residues, that retain BMP
activity. .
[039] As used herein, the term "conservative amino acid substitution"
refers to the replacement of a native amino acid by another amino acid of the
same or similar physicochemical properties. For example, substitution of one
acidic amino acid for another acidic amino acid (e.g., replacing an aspartic acid
residue with a glutamic acid residue), one basic amino acid for another basic
amino acid (e.g., replacing a histidine residue with a lysine residue), or one polar,
uncharged amino acid for another polar, uncharged amino acid (e.g., replacing a
serine residue for a threonine residue). The possibility and potential utility of
conservative amino acid substitutions in a protein of known sequence is well-
understood by one of ordinary skill in the art.
[040] As used herein, the "ED50" (effective dose 50%) is the amount of a
compound required to produce a specified effect in 50% of an animal population.
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As used herein, the "IC50" (inhibitory concentration 50%) is the concentration of a
compound which achieves 50% inhibition of its target.
[041]' As used herein, the term "immune cell" includes cells that are of
hematopoietic origin and that play a role in the immune response, as well as any .
epithelial or mesenchymal antigenrpresenting cells. Immune cells include B
lymphocytes, T lymphocytes; natural killer cells and myeloid cells,.such as
monocytes, macrophages, eosinophils, mast cells, basophils, and granulocytes.
[042] As used herein, the term "immune response" or "immune system
activity" includes T and/or B lymphocyte responses, i.e., cellular and/or humoral
immune responses. The immune response of a subject can be determined by, for
example, assaying antibody production, immune cell proliferation, the release of
cytokines, the expression of cell surface markers, cytotoxicity, or by monitoring
other indicators of immune system activity. . .
[043] The term "in combination" as used herein means that.a BMP
composition containing at least one BMP and a second therapeutic composition
are given either simultaneously or sequentially. If given sequentially, at the onset
of administration of the second compound, the first of the two compounds may still
be detectable at effective concentrations at the site of treatment.. It will be
understood that, if two therapeutic compositions are administered sequentially, the
interval between their administration will be determined by the exigencies of the
therapeutic situation and the experience of the treating physician.
[044] The methods disclosed herein can employ either a short,
intermediate, or extended course of therapy. As used herein.the language
"short course of therapy" includes a therapeutic regimen that is of relatively
short duration relative to the course of the illness being treated. For example, a
short course of therapy may last between about one to about four weeks. In .-...
contrast, an "intermediate course of therapy" includes a therapeutic regimen
that is of longer duration than a short course of therapy. For example, an
intermediate course of therapy can last from more than one month to about six
months (e.g., between about five to about 26 weeks). An "extended course of
therapy" includes those therapeutic regimens that last longer than about six
months, e.g.., from about seven months on. For example, an extended course of
therapy may last from about seven months to as long as the illness persists. The
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appropriateness of one or more of the courses of therapy described above for any
one individual can readily be determined by one of ordinary skill in the art. In
addition, the treatment appropriate for a subject may be changed over time as
required.
[045] In.the course of therapy, doses can be administered-early or late.
As used herein, "early dosing" includes a therapeutic regimen where BMP
compositions are administered to a patient at the onset of disease, e.g., at the
onset of clinical symptoms. Alternatively, "late dosing" includes a therapeutic
regimen where BMP compositions are administered after disease onset, e.g., after
diagnosis or establishment of the disease. . .
[046] As used herein, the term "TGF-β protein superfamily" refers to a
family of structurally-related growth factors. This family of related growth factors is
well-known in the art. Kingsley et al., "The TGF-β superfamily: new members,
new receptors, and new genetic tests of function in different organisms," Genes
Dev. 8:133-146 (1994); Hoodless et al., "Mechanism and function of signaling by
the TGF-β superfamily," Cure. Topics Microbiol. Immunol. 228:235-272 (1998).
The TGF-β superfamily includes bone morphogenetic proteins (BMPs), activin,
inhibin, mullerian-inhibiting substance, glial-derived neurotrophic factor, and a still
growing number of growth and differentiation factors (GDFs), such as GDF-8
(myostatin). Piek et al., "Specificity, diversity, and regulation in TGF-β superfamily
signaling," FASEB J. 13:2105-2124 (1999).
[047] As used herein, a "therapeuticaily effective amount" of at least
one BMP is an amount sufficient to stimulate increased hair growth, or to inhibit
immune system activity. That amount will depend upon the type and severity of
the disorder being treated. The optimal dose of BMP given may even vary in the
same patient depending upon the time at which it is administered.'
I. BONE MORPHOGENETIC PROTEIN COMPOSITIONS
'. [048] • According to the present invention, compositions are provided for
patients who exhibit signs of the autoimmune disease alopecia areata, other hair
loss disorders, or a variety of other autoimmune conditions. Some compositions
of the present invention are prepared by mixing at least one active agent with a
variety of pharmaceutically acceptable carriers and/or optional excipientsto form a
liquid, gel, or cream for topical (e.g., transdermal) application. Other compositions
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WO 2006/105359 PCT/US2006/011799
of the present invention are prepared by mixing at least one active agent with a
variety of pharmaceutically acceptable carriers and optional excipients to form a
liquid, gel or solid for administration by injection (e.g.; intraosseously,
intravenously, parenterally, or percutaneously). In some embodiments of the
invention, immune cells may be extracted from a patient suffering from a hair loss
disorder or an autoimmune disease, treated with the compositions of the
invention, and re-injected into the patient.
A. BONE MORPHOGENETIC PROTEINS
[049] The active agent may be selected from the family of proteins known
as the transforming growth factor-beta (TGF-β) superfamily of proteins, which
includes the activins, inhibins, and bone morphogenetic proteins (BMPs). In one
embodiment, the active agent includes at least one protein selected from the
subclass of proteins known generally as BMPs. The first BMPs (BMPs-1-4) were
identified by their ability to induce new bone formation in muscle tissue. Urist et
al., "Bone Formation By Autoinduction" Science 150:893-99 (1965). Additional
members of the protein subfamily were cloned based on homology with the
sequences of BMPs 1-4.
[050] BMPs have been shown to possess a wide range of growth and
differentiation activities, including induction of the growth and differentiation of
bone, connective, kidney, heart, and neuronal tissues. Rengachary, "Bone
Morphogenetic Proteins: Basic Concepts" Neurosurg. Focus 13(6): 1-6 (2002).
See, for example, descriptions of BMPs in the following publications: BMP-2,
BMP-3,. BMP-4, BMP-5, BMP-6, and BMP-7 (disclosed, for example, in. U.S.
Patent Nos. 5,013,649 (BMP-2 and B.MP-4); 5,116,738 (BMP-3); 5,106,748 (BMP-
5); 5,187,076 (BMP-6); and 5,141,905 (BMP-7)); BMP-8 (disclosed in PCT WO
91/18098); BMP-9 (disclosed in PCT WO 93/00432); BMP-10 (disclosed in PCT
WO 94/26893); BMP-11 (disclosed in PCT WO 94/26892); BMP-12 and BMP-13
(disclosed in PCT WO 95/16035); BMP-15 (disclosed iri U.S. Patent No.
5,635,372); BMP-1.6 (disclosed in U.S. Patent No. 6,331,612); MP52/GDF-5
(disclosed in PCT WO 93/16099); and BMP-17 and BMP-18 (disclosed in U.S.
Patent No. 6,027,917). Other TGF-B proteins that may be useful as the active
agent in the present invention include Vgr-2, Jones et.al., "Isolation of Vgr-2, a
novel member of the transforming growth factor-β-related gene family," Mol.
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WO 2006/105359 PCT/US2006/011799
Endocrinol. 6:1961-1968 (1992), and any of the growth and differentiation factors
(GDFs), including those described in PCT applications WO 94/15965; WO
94/15949; WO 95/01801; WO 95/01802; WO 94/21681; WO 94/15966; WO
95/10539; WO 96/01845; WO 96/02559 and others.
[051] A subset of BMPs that may be used in certain embodiments of the
present invention includes BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-
8, BMP-9, BMP-10, BMP-11, BMP-12 and BMP-13. In an illustrative embodiment,
the active agent is BMP-2, the sequence of which is disclosed in United States
Patent No. 5,013,649. In another exemplary embodiment, the active agent is
BMP-4, the sequence of which is also disclosed in United States Patent No.
5,013,649. In another embodiment, the composition contains two active agents, .
for example, BMP-2 and BMP-4. Other BMPs and TGF-β proteins known in the
art may also be used.
[052] The active agent may be recombinantly produced, or purified from
another source. The active agent, if a TGF-β protein such as a BMP, or other
dimeric protein, may be homodimeric, or may be heterodimeric with other BMPs
(e.g., a heterodimer composed of one monomer each of BMP-2 and BMP-5) or
with other members of the TGF-β superfamily, such as activins, inhibins and TGF-
β1 (e.g., a heterodimer composed of one monomer each of a BMP and a related
member of the TGF-β superfamily). Examples of such heterodimeric proteins are
described, for example in published PCT Patent Application WO 93/09229.
B. PHARMACEUTICAL FORMULATIONS AND ROUTES OF
ADMINISTRATION
[053] A pharmaceutical composition comprising at least one BMP may
contain a pharmaceutically acceptable carrier to render the composition suitable
for administration to a subject, and a therapeutically effective amount of the active
agent. The term subject is intended to include living organisms in which hair
growth or an immune response can be elicited, e.g., mammals. Examples of
subjects include, but are not limited to, humans, dogs, cats, mice, rats, and
transgenic species thereof. The at least one BMP can be administered by various
routes well known to persons of ordinary skill in the art, including, but not limited
to, parenterally, intravenously, percutaneously, intraosseously, or extra- .
corporeally.
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WO 2006/105359 PCT/US2006/011799
[054] A pharmaceutical composition for injection could be made up to
contain 5 ml sterile buffer containing 0.5% sucrose, 2.5% glycine, 5 mM L-
glutamic acid, 5 mM NaCI, and 0.01% polysorbate 80, at pH 4.50, and 5 mg of
BMP-2, for a final concentration of 1 mg/mi. A typical pharmaceutical composition
for intravenous infusion could be made up to contain 250 ml of sterile Ringer's
solution, and 0.05 to 500 mg of at least one BMP. Actual methods of preparing
compositions for administration by a variety of routes, including intraosseously,
intravenously, parenterally, or percutaneously will be known or apparent to those
skilled in the art and are described in more detail in, for example, PHILADELPHIA
COLLEGE OF PHARMACEUTICAL SCIENCES, REMINGTON'S PHARMACEUTICAL SCIENCES
(18th ed. 1990).
[055] Solutions or suspensions used for intraosseous, intravenous,
parenteral, or percutaneous, application typically include one or more of the
following components: a sterile diluent such as water for injection, saline solution,
fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic
solvents; antibacterial agents such as benzyl alcohol or methyl parabens;
antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as
ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates or
phosphates; and agents for the adjustment of tonicity such as sodium chloride or
dextrose. The pH can be adjusted with acids or bases, such as hydrochloric acid
or sodium hydroxide. Such preparations may be enclosed in ampoules,
disposable syringes or multiple.dose vials made of glass or plastic.
[056]. Pharmaceutical compositions suitable for injection include sterile
aqueous solutions or dispersions and sterile powders for the extemporaneous
preparation of sterile injectable solutions or dispersions. For intravenous
administration, suitable carriers include physiological saline, bacteriostatic water,.
Cremophor® EL (BASF, Parsippany, NJ) or phosphate buffered saline (PBS). In
all cases, the composition must be sterile and should be sufficiently fluid that it is
easily injectable via a standard sterile, disposable syringe. It 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.
[057] The carrier can be a solvent or dispersion medium containing, for
example, water, ethanol, polyol (e.g., glycerol, propylene glycol, or liquid
16

WO 2006/105359 FCT/US2006/011799
polyetheylene glycol, and the like), and suitable mixtures thereof. The proper
fluidity can be maintained, for example, by the use of a coating such as lecithin, by
the maintenance of the required particle size in the case of dispersion, and by the
use of surfactants. Prevention of the action of microorganisms can be achieved
by various antibacterial and antifungal agents, for example, parabens,
chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it
will be preferable to include isotonic agents, for example, sugars, polyalcohols .
such as mannitol or sorbitol, or sodium chloride in the composition. Prolonged
absorption of the injectable compositions can be brought about by including in the
composition an agent which delays absorption, for example, aluminum
monostearate or gelatin.
[058] In another illustrative embodiment, the compositions that stimulate
hair growth or inhibit immune system activity are prepared with carriers that will
protect the compound against rapid elimination from the body, such as a
controlled release formulation, including implants and microencapsulated delivery
systems. Biodegradable, biocompatible polymers can be used, such as ethylene
vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and
polylactic acid. Methods for preparation of such formulations will be apparent to .
those skilled in the art. The materials can also be obtained commercially from
Alza Corporation (Mountain View, CA) or from other commercial suppliers.
Liposomal suspensions may also be used as pharmaceutically acceptable
carriers. Such suspensions may be targeted to specific subpopulations of cells or
specific subcellular compartments, and can be prepared according to methods
known to those skilled in the art, for example, as described in U.S. Patent No.
4,522,811.
II. METHODS OF TREATMENT
A. CONDITIONS TO BE TREATED
[059] Subjects suffering from a wide variety of disorders may be treated by
administering the compositions of the invention. For example, the present
invention provides methods of treatment for alopecia areata, as well as the related
disorders alopecia totalis and alopecia universalis. The invention also provides
methods of treatment for other hair loss disorders, including, but not limited to,
androgenic alopecia (affecting both male and female patients), telogen effluvium,
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WO 2006/105359 PCT/US2006/011799
anagen effluvium, and chemotherapy-induced hair loss. The compositions of the
invention may be administered alone, or in combination with additional
compounds well known to one of ordinary skill in the art that are used to treat
other pathologic conditions or. diseases resulting in hair loss.
[060] The present invention further provides methods of treatment for a
wide variety of immune system disorders, including, but not limited to, ankylosing
spondylitis, antiphospholipid syndrome, Addison's Disease, autoimmune
hemolytic anemia, autoimmune hepatitis, autoimmune lymphoproliferative
syndrome (ALPS), autoimmune thrombocytopenic purpura (ATP), Behcet's
Disease, bulbous pemphigoid, cardiomyopathy, celiac disease, Dermatitis
Herpetiformis, chronic fatigue immune dysfunction syndrome (CFIDS), chronic
inflammatory demyelinating polyneuropathy (CIDP), cicatricial pemphigoid,
systemic sclerosis (CREST Syndrome), cold agglutinin disease, Crohn's Disease,
cutaneous vasculitis, Degos1 Disease, dermatomyositis, juvenile dermatomyositis,
discoid lupus erythematosus, essential mixed cryoglobulinemia, fibromyalgia,
Goodpasture's Syndrome, Graves' Disease, Guillain-Barre Syndrome,
Hashimoto's Thyroiditis, idiopathic pulmonary fibrosis, idiopathic
thrombocytopenia purpura (ITP), immunogiobulin A nephropathy, insulin-
dependent diabetes mellitus, juvenile arthritis, Kawasaki's Disease, lichen planus,
membranous glomerulonephritis, Meniere's Disease, mixed connective tissue
disease, multifocal motor neuropathy, multiple sclerosis, myasthenia gravis,
pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polychondritis,
.polyglandular syndrome, polymyalgia rheumatica, polymyositis and
dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis,
psoriasis, Raynaud's Phenomenon, Reiter's Syndrome, rheumatic fever,
rheumatoid arthritis, sarcoidosis, scleroderma, Sjogren's Syndrome, Stiff-Man
Syndrome, systemic lupus erthryomatosus, Takayasu arteritis, temporal
arteritis/giant cell arteritis, ulcerative colitis, uveitis, vasculitis, vitiligo, and
Wegener's Granulomatosis. The compositions of the invention may be
. administered alone, or in combination with additional compounds well known to
one of ordinary skill in the art to inhibit the immune response.
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WO 2006/105359 PCT/US2006/011799
B. MECHANISMS OF ACTION
[061] The BMP compositions of the invention may act in any or all of the
following ways to stimulate hair growth and/or to inhibit the immune system. The
BMP compositions of the invention may act upon a patient's population of
circulating T lymphocytes to alter existing equilibria and cause systemic changes
in the immune system, thereby inducing hair growth and/or ameliorating the
phenotype of an immune system disorder. The BMP compositions may also be
absorbing excess amounts of BMP antagonists, such as anti-BMP, antibodies,
noggin, chordin, and Cerberus proteins, or other BMP antagonists, to systemicaily
alter BMP metabolism or the activity of the antagonists themselves, thereby
inducing hair growth and/or ameliorating the phenotype of an immune system
disorder. Finally, the BMP compositions of the invention may increase circulating
concentrations of a BMP throughout a subject's body to levels sufficient
systemicaily to stimulate hair growth and/or ameliorate the phenotype of an
immune system disorder.
C. RANGE OF DOSAGE
[062] It is advantageous to formulate compositions administered by
injection or inhalation in dosage unit form for ease of administration and uniformity
of dosage. Dosage unit form as used herein refers to physically discrete units
prepared as unitary dosages for the mammalian subjects to be treated, with each
unit containing a predetermined quantity of active compound calculated to.
produce the desired therapeutic effect in association with the required
pharmaceutical carrier; .
[063] Data obtained from cell culture.assays and animal studies can be
used in formulating a range of dosage for use in humans. The dosage may vary
within this range depending upon the dosage form employed and the route of.
administration utilized. For any compound used in the method of the invention,
the therapeutically effective dose can be estimated initially from cell culture
assays and dosages refined to more accurately determine useful, therapeutically
effective doses in humans,
[064] In an exemplary embodiment of the present invention, a
therapeutically effective amount of at least one of BMP-2, BMP-3, BMP-4; BMP-5,
BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13, BMP-14,
19

WO 2006/105359 PCT/US2006/011799
BMP-15, BMP-16, BMP-17, BMP-18, or MP52/GDF-5 may be administered to a
subject. In another embodiment, a therapeutically effective amount of at least one
of BMP-2, BMP-4, BMP-5, BMP-6, BMP-7, BMP-10, BMP-12 orBMP-13 maybe
administered to a subject. In still another embodiment, a therapeutically effective
amount of at least one of BMP-2 or BMP-4 may be administered to a patient. In
another embodiment, a therapeutically effective amount of both BMP-2 and BMP-
4 may be administered to a subject. The optimal dose of at least one BMP given
may even vary in the same patient depending upon the time at which it is
administered. Generally, the amount of protein to be delivered may range from
about 0.05 to about 500 mg, from about 0.5 to about 50 mg, from about 1 to about
25 mg, and from about 5 to about 10 mg per dose. In general, a therapeutically
effective amount of at least one BMP would be delivered in solution at a
concentration from about 0.001 mg/ml to about 100 mg/ml, from about 0.01 mg/ml
to about 50 mg/ml, from about 0.1 mg/m! to about 25 mg/ml, and from about 1
mg/ml to about 5 mg/ml.
[065] The specific dose can be readily calculated by one of ordinary skill in
the art, according to the approximate body weight or volume, or body surface area
of the patient. The dose will also be calculated dependent upon the particular
route of administration selected. Further refinement of the calculations necessary
to determine the appropriate dosage for treatment is routinely made by those of
ordinary skill in the art. Such calculations can be made without undue
experimentation by one skilled in the art, based upon standard dose-response
studies. It will be understood that the amount of the composition actually
administered will be determined by a practitioner, in light of relevant
circumstances including the condition or conditions to be treated, the choice of
composition to be administered, the age, weight, and response of the individual
patient, the severity of the patient's symptoms, and the chosen route of
administration.
[066] Toxicity and therapeutic efficacy of such compounds can be
determined by standard pharmaceutical procedures in cell cultures or
experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the
population) and the ED50 (the dose therapeutically effective in 50% of the
population). The dose ratio between toxic and therapeutic effects is the
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WO 2006/105359 PCT/US2006/011799
therapeutic index, which can be expressed as the ratio LD5O/ED50. Compounds
which exhibit large therapeutic indices are preferred. While compounds that
exhibit toxic side effects may be used, care should be taken to design a delivery
system that targets such compounds to the site of affected tissue in order to
minimize potential damage to uninfected cells, thereby reducing side effects.
[067] The data obtained from cell culture assays and/or animal studies
can be used in formulating a range of dosage for use in humans. The dosage of
such compounds lies preferably within a range of circulating concentrations that
include the ED50 with little or no toxicity. The dosage may vary within this range
depending upon the dosage form, employed and the route of administration
utilized. For any compound used in the methods of the invention, the
therapeutically effective dose can be estimated initially from cell culture assays,
and dosages refined without undue experimentation to more accurately determine
useful doses in humans. For example, plasma levels of the active agent may be
measured by high performance liquid chromatography.
[068] Thus, the dosage of any of the subject agents, can be easily
determined by one of ordinary skill in the art. The dose may vary depending on
the age, health and weight of the recipient, the extent of disease, kind of •
concurrent treatment, if any, frequency of treatment and the nature of the effect
desired. It should be noted that the dose of BMP given to one subject may vary
during the course of the treatment.
P. TIMING OF TREATMENT
[069] The skilled artisan will appreciate that certain factors may influence
the dosage required to effectively treat a subject, including but not limited to the
severity of the disease or disorder, previous, treatments, the general health and/or
age of the subject, and other diseases present. Moreover, treatment of a subject
can include a single treatment or can include a series of treatments administered
daily, weekly, monthly, bi-monthly, tri-monthly, bi-annually, and annually. In some
cases it may be necessary to continue treatment for an indefinite period, or for as
long as hair growth or a decreased immune response is desired. It will also be .
appreciated that the therapeutically effective dosage of a BMP used to stimulate
hair growth or to inhibit the immune system may increase or decrease over the
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WO 2006/105359 PCT/US2006/011799
course of a particular treatment. Changes in dosage may result from relapse of
the disease or.worsening of symptoms.
[070] The present compositions comprising BMPs can be administered for
prophylactic and/or therapeutic treatments, in prophylactic application,
compositions can be administered to a patient especially susceptible or otherwise
expecting to suffer hair loss or increased immune system activity, for example,
resulting from a course of chemotherapy. In therapeutic application, compositions
can be administered to a patient already suffering from a disease, for example,
alopecia areata, alopecia totalis, alopecia universalis, androgenic alopecia,
rheumatoid arthritis or other autoimmune disease, in an amount sufficient to
ameliorate the symptoms of the disease and associated complications. Single or
multiple administrations of the compositions can be carried out with dose levels.
and pattern sufficient effectively to treat the patient being selected by the treating
physician. -
E. COMBINATION THERAPY FOR STIMULATING HAIR GROWTH
[071] Administration of the compositions of the invention as described
herein may be as a therapeutically effective formulation containing a
therapeutically active amount of at least one BMP alone or in combination with
any other therapeutic composition or molecule. The combination therapy is useful
for treating pathological conditions or disorders resulting in hair loss. The term "in
combination" in this context means that the BMP composition and a second
therapeutic composition are given either simultaneously or sequentially. If given
sequentially, at the onset of administration of the. second compound, the first of
the two compounds may still be detectable at effective concentrations at the site
of treatment.
[072]. For example, the combination therapy may include at least one BMP
composition co-formulated with, and/or co-administered with, at least one
additional therapeutic agent for stimulating hair growth. The additional agents
may include at least one of the following, administered either orally, topically, by
inhalation or by injection: (1) corticosteroids, such as prednisone,
dexamethasone, or hydrocortisone; (2) calcineurin inhibitors known to have
immunosuppressive activity, such as cyclosporin A, pimecrolimus or tacrolimus;
(3) minoxidil; or (4) finasteride. Such combination therapies may advantageously
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WO 2006/105359 PCT/US2006/011799
utilize lower dosages of the administered therapeutic agents, thus avoiding
possible toxicities or complications associated with the various monotherapies
alone. Moreover, the additional therapeutic agents disclosed herein act upon
metabolic pathways other than those regulating BMP metabolism, and thus are
expected to enhance and/or synergize with the effects of the BMP compositions.
[073] Administration of a therapeutically active amount of the compositions
of the present invention is defined as an amount effective, at dosages and for
periods of time necessary to achieve the desired result. For example, a.
therapeutically active amount of at least one BMP, a corticosteroid, a calcineurin
inhibitor, minoxidil, or finasteride may vary according to factors such as the
disease state, age, sex, and weight of the individual, and the ability of the
compound to elicit a desired response in the individual. A dosage regime may be
adjusted to provide the optimum therapeutic response. For example, several
divided doses may be administered daily, a single dose may be administered
daily, weekly, monthly or at longer intervals, or the dose may be proportionally
reduced as indicated by the exigencies of the therapeutic situation.
F. COMBINATION THERAPY FOR INHIBITING IMMUNE SYSTEM
ACTIVITY
[074] Administration of the compositions of the invention as described
herein can be as a therapeutically effective formulation containing a
therapeutically active amount of at least one BMP alone or in,combination with
any other therapeutic composition or molecule. The combination therapy is useful
for treating pathological conditions or disorders of the immune system, especially
those characterized by aberrant autoimmune activity. The term "in combination"
in this context means that the BMP composition and a second therapeutic
composition are given either simultaneously or sequentially. If given sequentially,
at the onset of administration of the second compound, the first of the two
compounds may still be detectable at effective concentrations at the site of
treatment.
[075] For example, the combination therapy can include at least one BMP
composition co-formulated with, and/or co-administered with, at least one
additional therapeutic agent for reducing immune system activity. The additional
agents may include at least one of the following, administered either orally,
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WO 2006/105359 PCT/US2006/011799
topically, by inhalation or by injection: (1) calcineurin inhibitors known to have
immunosuppressive activity, such as cyclosporin A, pimecrolimus or tacrolimus; or
(2) other compounds with immunosuppressive activity, for example, azathioprine,
rnycophenoiate rnofeti!, raparp.ycin or rapamycin analogs (e.g., CCI-779).
[076] Administration of a therapeutically active amount of the compositions
of the present invention is defined as an amount effective, at dosages and for
periods of time necessary to achieve the desired result. For example, a
therapeutically active amount of at least one BMP, a calcineurin inhibitor, or
another immunosuppressive compound may vary according to factors such as the
disease state, age, sex, and weight of the individual, and. the ability of the
compound to elicit a desired response in the individual. A dosage regime may be
adjusted to provide the optimum therapeutic response. For example, several
divided doses may be administered daily, a single dose may be administered
daily, weekly, monthly or at longer intervals, or the dose may be proportionally
reduced as indicated by the exigencies of the therapeutic situation.
[077] The BMP compositions disclosed herein can be used in combination
with other therapeutic agents to treat specific immune disorders as discussed in
further detail below.
[078] Non-limiting examples of agents for treating arthritic disorders (e.g.,
rheumatoid arthritis, inflammatory arthritis, rheumatoid arthritis, juvenile .
rheumatoid arthritis, osteoarthritis and psoriatic arthritis), with which a BMP
composition may be combined include at least one of the following: TNF
antagonists (such as anti-TNF antibodies); soluble fragments of TNF receptors
(e.g., human p55 and p75) and derivatives thereof (such as p55 kdTNFR-lgG (55
kD TNF receptor-IgG fusion protein, Lenercept®) and 75 kd TNFR-IgG (75 kD
TNF receptor-IgG fusion protein, Enbrel®)); TNF enzyme antagonists ^such as
TNFa converting enzyme, orTACE, inhibitors); antagonists of 1L-12, IL-15, IL-17,
IL-18, and IL-22; T cell and B cell depleting agents (such, as anti-CD4 or anti-
CD22 antibodies); small molecule inhibitors (such as methotrexate and
leflunomide); COX-2 and cPLA2 inhibitors; non-steroidal anti-inflammatory drugs
(NSAIDs); p38, TPL-2, Mk-2, and NFKB inhibitors; receptor for advanced glycation
end products (RAGE) or soluble RAGE; P-selectin or PSGL-1 inhibitors (such as
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WO 2006/105359 PCT/US2006/011799
small molecule inhibitors and antibodies thereto); estrogen receptor (3 (ER(3)
agonists, and ERβ-NFkB antagonists.
[079] Non-limiting examples of agents for treating multiple sclerosis with
which a BMP composition may be combined include interferon-β (for example,
IFNβ-1a and IFNP-1b), copaxone, corticosteroids, IL-I inhibitors, TNF inhibitors,
antibodies to CD40 ligand, antibodies to CD80, and IL-12 antagonists.
[080] Non-limiting examples of agents for treating inflammatory bowel
disease or Crohn's disease with which BMP compositions may be combined
. include budenoside; epidermal growth factor; corticosteroids; cyclosporin;
sulfasalazine; aminosalicylates; 6-mercaptopurine; azathioprine; metronidazoie;
lipoxygenase inhibitors; mesalamine; olsalazine; balsalazide; antioxidants;
thromboxane inhibitors; IL-1 receptor antagonists; anti-IL-1 monoclonal
antibodies; anti-IL-6 monoclonal antibodies; growth factors; elastase inhibitors;
pyridinyl-imidazole compounds; TNF antagonists as described herein; IL-4, IL-10,
IL-13, and/or TGFb or agonists thereof (e.g., agonist antibodies); IL-11;
glucuronide- or dextran-conjugated prodrugs of prednisolone, dexamethasone or
budesonide; ICAM-1 antisense phosphorothioate oligodeoxynucleotides (ISIS
2302; Isis Pharmaceuticals, Inc.); soluble complement receptor 1 (TP10; T Cell
Sciences, Inc.); slow-release mesalazine; methotrexate; antagonists of Platelet
Activating Factor (PAF); ciprofloxacin; and lignocaine.
G. METHODS FOR EVALUATING THE ABILITY OF
PHARMACEUTICAL COMPOSITIONS TQ STIMULATE HAIR
GROWTH
[081J The ability of the compositions of the invention as described herein
to stimulate hair growth in a subject can be determined by, for example, assaying
expression of hair- or hair follicle-specific markers of cell growth and
differentiation, including keratin 6; keratin 16, keratin 17, (3-catenin, and
trichohyalin, by measuring uptake of bromodeoxyuridine in dividing hair follicle
stem cells or by measuring alkaline phosphatase activity in dermal papilla cells.
The compositions of the invention may also be tested for their ability to stimulate
hair growth in vitro on cultures of matrix keratinocytes, on dissected human hair
follicles, or on whole skin explants grown on collagen sponges for the ability to
induce cell and/or hair growth and proliferation. Alternatively, the compositions of
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WO 2006/105359 PCT/US2006/011799
the invention may be tested for their ability to stimulate hair growth on mice after
clipping or shaving a portion of their body hair, as described in Example 2.
[082] It will be understood that each of the various hair loss disorders •
treatable by the methods of the present invention has a characteristic molecular
phenotype measurable using techniques well known to persons of ordinary skill in
the art. That phenotype may be characterized by increased or decreased levels
of circulating auto-antibodies, increased or decreased levels of T lymphocytes or
subpopulations thereof, altered expression levels or patterns of hair follicle-
specific cell surface antigens, or by other molecular markers of cell growth and
proliferation. These phenotypes may be assayed before and after treatment with
the compositions of the invention using a variety of routine methods, including flow
cytometry, immunohistochemistry, enzyme-linked immunosorbent assays
(ELISA), Western blotting, reverse transcription-polymerase chain reaction (RT-
PCR), and/or transcription profiling.
H. METHODS FOR EVALUATING THE ABILITY OF
PHARMACEUTICAL COMPOSITIONS TO INHIBIT IMMUNE
SYSTEM ACTIVITY .
[083] The ability of the compositions of the invention as described herein
to inhibit immune system activity in a subject can be determined by, for example,
assaying serum antibody levels, immune cell proliferation or markers thereof, the
release of cytokines, the expression of cell surface markers, cytotoxicity, or by
monitoring other indicators of immune system activity. It will be understood that
each of the various autoimmune disorders treatable by the methods of the present
invention has a characteristic molecular phenotype measurable using techniques
well known to persons of ordinary skill in the art. That phenotype may be
characterized by increased or decreased levels of circulating auto-antibodies,.
increased or decreased levels of T or B lymphocytes or subpopulations thereof,
altered expression levels or patterns of T lymphocyte- or B lymphocyte-specific
cell surface antigens, or by other molecular markers of cell growth and
proliferation. These phenotypes may be assayed before and after treatment with
the compositions of the invention using a variety of routine methods, including flow
cytometry, immunohistochemistry, enzyme-linked immunosorbent assays.
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WO 2006/105359 PCT/US2006/011799
(ELISA), Western blotting, reverse transcription-polymerase chain reaction (RT-
PCR), and/or transcription profiling.
[084] For example, patients suffering from systemic lupus erythematosus
(SLE) frequently have high serum levels of antibodies directed to DNA, or to
ribonucleoproteins (RNPs). The efficacy of the methods of treatment described
herein may be evaluated by quantifying the amount of circulating anti-DNA or anti-
RNP antibodies in the serum of a patient suffering from SLE before and after
treatment using, for example, a sandwich ELISA. A method of treatment as.
described herein would be deemed effective if it resulted in a decrease in
circulating levels of anti-DNA or anti-RNP auto-antibodies compared to pre-
treatment levels.
EXAMPLES
EXAMPLE 1: TREATING A 21-YEAR-OLD MAN HAVING ALOPECIA
UNIVERSALIS WITH BMP-2
[085] A 21-year-old man presented to the dermatology department of
Norfolk & Norwich University in March, 2004, with alopecia. The patient had first
noticed patches of scalp hair loss 2 years previously. Hair loss progressed rapidly
from this first observation, and within 6 months the patient had lost every hair from
all body sites. The patient's past medical history includes asthma and childhood
eczema, as well as a family history of alopecia areata (both his maternal
grandmother and one cousin had the disease). Examination confirmed the
diagnosis of alopecia universalis in an otherwise fit young man. Routine blood
tests, including full blood count, urea and electrolytes, liver function tests and
auto-immune serology, were all within normal range.
[086] One month later, the patient fractured his left tibia while playing
soccer. His tibia was set with a Russell-Taylor tibial nail inserted/Six days later,
on April 25, 2004, the patient consented to participate in a trial using recombinant
human BMP-2 (rhBMP-2), potentially to speed the healing of the tibial fracture.
The closed tibial fracture was injected with 5 ml of rhBMP-2 at 1.0 mg/ml in a
calcium phosphate matrix, and the patient was subsequently discharged from
hospital. Six weeks later, the patient noticed hair re-growth initially on his scalp
(see Figure 1). Hair growth became more extensive, with hair eventually returning
to eyebrows, axillae, arms and pubic area. This hair re-growth was maintained for
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WO 2006/105359 PCT/US2006/011799
six months following the treatment with rhBMP-2. The tibia! fracture has also
healed satisfactorily.
EXAMPLE 2: BMP-2 AND BMP-4 STIMULATE HAIR GROWTH IN C57BL/6
MICE
[087] The ability of the compositions of the invention to stimulate hair
growth is evaluated using a population of 45 sex- and age-matched C57BL/6
mice, separated into three groups of 15: two experimental groups and one control
group. Hair from a roughly 4-cm2 region of dorsal skin on all animals is clipped to
0.1 mm in length. The experiment will test a range of five different doses of BMP-
2 and BMP-4: 500 ng, 1 μg, 2.5 μg, 5 μg, 10 μg. Each single use dose is
prepared in an isotonic saline solution at BMP-2 or BMP-4 concentrations
sufficient to allow each dose to be administered in a total voiume of 25 μl.
[088] Three animals in the first experimental group, are injected with 25 μl
each of isotonic saline containing 500 ng of BMP-2 (20 ng/μl), three are injected
with 25 μl each of isotonic saline containing 1 μg of BMP-2 (40 ng/μl), three are
injected with 25 μl each of isotonic.saline containing 2.5 μg of BMP-2 (100 ng/μl),
three are injected with 25 μl each of isotonic saline containing 5 μg of BMP-2 (200
ng/μl), and three are injected with 25 μl each of isotonic saline containing 10 μg of
BMP-2 (400 ng/μl). Three animals in the second experimental group are injected
with 25 μl each of isotonic saline containing 500 ng of BMP-4 (20 ng/μl), three are
injected with 25 μl each of isotonic saline containing 1 μg of BMP-4 (40 ng/μl),
three are injected with 25 μl each of isotonic saline containing 2.5 μg of BMP-4
(100 ng/μl), three are injected with 25 μl each of isotonic saline containing 5 μg of
BMP-4 (200 ng/μl), and three are injected with 25 μl each of isotonic saline
containing 10 μg of BMP-4 (400 ng/μl).
[089] Similarly, animals in the control group are separated into five groups
of three. The first group of three is injected with 25 μl each of an isotonic saline
solution containing 500 ng bovine serum albumin (BSA) (20 ng/μl), the second
group is injected with 25 μl each of an isotonic saline solution containing 1 μg BSA
(40 ng/μl), the third group is injected with 25 μl each of an isotonic saline solution
containing 2.5 μg BSA (100 ng/μl), the fourth group is injected with 25 μl each of
an isotonic saline solution containing 5 μg BSA (200 ng/μl), and the fifth group is
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WO 2006/105359 PCT/US2006/011799
injected with 25 μl| each of an isotonic saline solution containing 10 μg BSA (400
ng/μl).
[090] All mice in both groups are examined twice daily for hair growth for a
period of twenty-one days. It is expected that hair regrowth will be faster in mice
receiving BMP-2 or BMP-4.than in mice receiving only BSA. It is further expected
that the stimulation of hair regrowth will correlate with the amount of BMP-2 or
BMP-4 received. Thus it is expected that the difference in hair regrowth between
the control group and the experimental groups will be most significant and
noticeable for those mice receiving either 5 μg (200 ng/μl) or 10 μg (400 ng/μl) of
BMP-2 or BMP-4. Similarly, it is expected that the difference in hair regrowth will
be least significant and noticeable for those mice receiving either 500 ng (20
ng/μl) or 1 μg (40 ng/μl) of BMP-2 or BMP-4. . .
[091] After twenty-one days, the mice are euthanized and skin sections"
are prepared for histologica! and immunohistochemical analysis of markers of hair
follicle growth and development, including bromodeoxyuridine (BrdU) labelling,
triohohyalin and hair keratin expression, and alkaline phosphatase activity in
dermal papilla cells. It is expected that BrdU labeling, a marker of proliferation in
epidermal stem cells of the hair follicle, will increase with increasing doses of
BMP-2 or BMP-4. Similarly, expression of hair follicle'Specific keratins and
trichohyalin is expected to increase with increasing doses of BMP-2 or BMP-4, as
measured by real time RT-PCR and transcription profiling.
EXAMPLE 3: TREATING PATIENTS SUFFERING FROM ANDROGENIC
ALOPECIA WITH BMP-2
[092] In one study, a cohort of normal, healthy men suffering from
andrqgenic alopecia is identified. Initially, all subjects are evaluated by routine
blood tests, including full blood count, urea and electrolytes, blood pressure and
liver function tests as well as auto-immune serology. In accord with generally
accepted practice for the conduct of a standard randomized, double-blind clinical
trial, the patients are given identification numbers separating them into an
experimental group that will receive BMP-2, and a control group that will receive a
placebo. The trial will test three treatment regimens, ranging from a single
injected dose of 5 mg, to three bi-monthly injections of 5 mg each, to six
consecutive monthly injections of 5 mg each. Whether administered alone or in
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WO 2006/105359 . PCT/US2006/011799
combination (see below), the BMP-2 will be administered by a single injection of 5.
ml of BMP-2 prepared at a concentration of 1 mg/ml, formulated in an appropriate
sterile pharmaceutical solution. Outcome assessments are based on weekly
visual examinations of hair growth and.density, as we!! as on levels of markers of
hair follicle growth and development measured in tissue taken from scalp biopsies
before treatment, and three, six and twelve months after treatment. It is expected
that increased hair growth and density, as well as increased expression of hair,
follicle-specific intermediate filament proteins such as keratins and trichohyalin,
will correlate directly with increased doses of BMP-2,
[093] In another study, a second cohort of normal, healthy men suffering
from androgenic alopecia is identified.' As before, all subjects are evaluated by
routine blood tests, including full blood count, urea and electrolytes, blood
pressure and liver function tests as well as auto-immune serology. In accord with
generally accepted practice for the conduct of a standard randomized, double-
blind clinical trial, the patients are given identification numbers separating them
into an experimental group that will receive BMP-2, and a control group that will
receive a placebo. The trial will test four combination therapies, each including
three bi-monthly injections of 5 mg BMP-2 combined with (1) prednisone, (2)
cyclosporin A, (3) a 5% topical solution of minoxidil, or (4) oral finasteride. Doses
of the latter four medications are administered in accord with accepted clinical
practice as determined by the physicians overseeing the trial. Outcome
assessments are based on weekly visual examinations of hair growth and density,
as well as on levels of markers of hair follicle growth and development measured
in tissue taken from scalp biopsies before treatment, and three, six and twelve
months after treatment. It is expected that increased hair growth and density, as
well as increased expression of hair follicle-specific intermediate filament proteins
such as keratins and trichohyalin, will correlate directly with increased doses of.
BMP-2 in combination with corticosteroids, calcineurin inhibitors, minoxidil or
finasteride.
EXAMPLE 4: TREATING PATIENTS SUFFERING FROM SYSTEMIC LUPUS
ERYTHEMATOSUS WITH BMP-2
[094] In one study, a cohort of normal, otherwise healthy female patients
suffering from systemic lupus erythematosus (SLE) is identified. Initially, all.
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WO 2006/105359 PCT/US2006/011799
subjects are evaluated by routine blood tests, including full blood count, urea and
electrolytes, blood pressure and liver function tests as well as auto-immune
serology. In accord with generally accepted practice for the conduct of a standard
randomized; double-blind clinical trial, the patients are given identification
numbers separating them into an experimental group that will receive BMP-2, and
a control group that will receive a placebo. The trial will test three treatment
regimens, ranging from a single injected dose of 5 mg, to three bi-monthly
injections of 5 mg each, to six consecutive monthly injections of 5 mg each.
Whether administered alone or in combination (see below), the BMP-2 will be
administered by a single injection of 5 ml of BMP-2 prepared at a concentration of
1 mg/ml, formulated in an appropriate sterile pharmaceutical, solution. Outcome
assessments are based on circulating serum concentrations of antibodies :
characteristic.of SLE, including anti-dsDNA (directed against double stranded
DNA), anti-Sm (directed against six different small, nuclear RNA molecules), and
anti-RNP (directed against U1 RNA), as measured by conventional ELISA before
treatment, and weekly for twelve months after treatment commences. It is
expected that a decrease in serum levels of some or all of these auto-antibodies
will correlate with increasing doses of BMP-2.
[095] In another study, a second cohort of normal, healthy women
suffering from SLE is identified. As before, all subjects are evaluated by.routine
blood tests, including full blood count, urea and electrolytes, blood pressure and
liver function tests as well as auto-immune serology. !n accord with generally
accepted practice for the conduct of a standard randomized, double-blind clinical
trial, the patients are given identification numbers separating them into an-
experimental group that will receive BMP-2 in combination with one of two types
of immunosuppressive compounds, and a control group that will receive a
placebo. The trial will test two combination therapies, each including three bi-
monthly injections of 5 mg BMP-2 combined with (1) cyclosporin A, or (2).
rapamycin. Doses of the latter two medications are administered in accord with
accepted clinical practice as determined by the physicians overseeing the trial.
Outcome assessments are based on circulating serum concentrations of
antibodies characteristic of SLE, including anti-dsDNA (directed against double
stranded DNA), anti-Sm (directed against six different small, nuclear RNA
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WO 2006/105359 PCT/US2006/011799
molecules), and anti-RNP (directed against U1 RNA), as measured by
conventional ELISA before treatment, and weekly for twelve months after
treatment commences. It is expected that a decrease in serum levels of some or
all of these auto-antibodies will correlate with increasing doses of BMP-2 in
combination with a calcineurin inhibitor or a known immunosuppressor.
[096] The specification is most thoroughly understood in light of the
teachings of the references cited within the specification. The embodiments within
the specification provide an illustration of embodiments of the invention and
should not be construed to limit the scope of the invention. The skilled artisan
readily recognizes that many other embodiments are encompassed by the
invention. All publications and patents cited in this disclosure are incorporated by
reference in their entirety. To the extent the material incorporated by reference
contradicts or is inconsistent with this specification, the specification will
supercede any such material. The citation of any references herein is not an
admission that such references are prior art to the present invention.
[097] Unless otherwise indicated, all numbers expressing quantities of
ingredients, reaction conditions, and so forth used in the specification, including
claims, are to be understood as being modified in a!! instances by the term
"about". Accordingly, unless otherwise indicated to the contrary, the numerical .
parameters are approximations and may vary depending upon the desired
properties sought to be obtained by the present invention. At the very least, and
not as an attempt to limit the application of the doctrine of equivalents to the. scope
of the claims, each numerical parameter should be construed in light of the
number of significant digits and ordinary rounding approaches. . .
[098] Unless otherwise indicated, the term "at least" preceding a series of
elements is to be understood to refer to every element in the series. Those skilled
in the art will recognize, or be able to ascertain using no more than routine
experimentation, many equivalents to the specific embodiments of the invention
described herein. Such equivalents are intended to be encompassed by the
following claims.
REFERENCES
[099] Bernstein, et al., "The aesthetics of follicular transplantation,"
Dermatol. Surg. 23(9):785-799 (1997).
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WO 2006/105359 PCT/US2006/011799
[0100] Botchkarev, V. A., "Bone morphogenetic proteins and their
antagonists in skin and hair follicle biology,"' J. Invest Dermatol. 120(1):36-47
(2002).
[0101] Botchkarev et a!., "BMP'signaling in the control of skin development
and hair follicle growth," Differentiation 72:512-526 (2004).
[0102] Brown et al., "A current review of medical therapy for benign
prostatic hyperplasia," J. Am. Osteopath. Assoc. 104(S2):S11-S16 (2004).
[0103] Cotsarelis et al., "Towards a molecular understanding of hair loss
and its treatment," TRENDS in Mol. Med. 7(7):293-301 (2001).
[0104] Dormois et al., "Minoxidil in severe hypertension: value when
conventional drugs have failed," Am. Heart J. 90:360-368 (1975).
[0105] Foley, P.A., "Recent advances: dermatology," Brit. Med. J. 320:850-
853(2000).
[0106] Hull et al., "Guidelines for the management of alopecia areata," Br.
J. Dermatol. 149:692-699 (2003).
[0107] Hoodless et al., "Mechanism and function of signaling by the TGF-S
superfamily," Curr. Topics Microbiol. Immunol. 228:235-272 (1998).
[0108] Jones et al., "Isolation of Vgr-2, a novel member of the transforming
growth factor-B-related gene family," Mol. Endocrinol. 6:1961-1968 (1992).
[0109] Kalish, et al., "Alopecia areata: autoimmunity—the evidence is
compelling," J. Invest. Dermatol. 8(2):164-167 (2003).
[0110] Kingsley et al., "The TGF-S superfamily: new members, new
receptors, and new genetic tests of function in different organisms," Genes Dev.
8:133-146(1994).
[0111] MacDonald, N., "Alopecia areata: identification and current
treatment approaches," Dermatol. Nurs. 11:356-359 (1999).
[0112] Messenger, A.G. et al., "Minoxidil: mechanisms of action on hair
growth," Brit. J. Dermatol. 150:186-194 (2004).
[0113] NATIONAL INSTITUTES OF HEALTH, Questions & Answers About
Alopecia Areata (2003).
[0114] Paus, et al., "The biology of hair follicles," N. Engl. J. Med.
341 (7):491 -497 (1999).
[0115] Physician's Desk Reference® (49th ed. 1995).
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[0116] Physician's Desk Reference® (58th ed. 2004).
[0117] Piek et al., "Specificity, diversity, and regulation in TGF-β
superfamily signaling," FASEB J. 13:2105-2124 (1999).
[0118] Philadelphia College of Pharmaceutical Sciences, Remington's
Pharmaceutical Sciences (18th ed. 1990).
[0119] Rengachary, "Bone Morphogenetic Proteins: Basic Concepts"
Neurosurg. Focus 13(6): 1-6 (2002).
[0120] Rosen et al., "Bone Morphogenetic Proteins" Principles of Bone
Biology 2:919-928 (2002).
[0121] Stenn et al., "Controls of Hair Follicle Cycling," Physiol. Rev. 81:449-
494(2001).
[0122] Urist et al., "Bone Formation By Autoinduction" Science 150:893-99
(1965).
[0123] Vierhapper et al., "Production rates of testosterone and of
dihydrotestosterone in female pattern hair loss," Metabolism 52(7):927-929
(2003). .
[0124] Wdzney, J.M., "Bone morphogenetic proteins and their gene
expression," CELLULAR AND MOLECULAR BIOLOGY OF BONE 131-167 (Noda, M. ed.
1993).
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WHAT IS CLAIMED IS:
1. A method of treating a hair loss disorder comprising:
(a) identifying a patient suffering from a hair loss disorder;
(b) administering a composition comprising a therapeutically
effective amount of at least one BMP chosen from the group consisting of BMP-2,
BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9..BMP-10, BMP-11, BMP-
12, BMP-13, BMP-14, BMP-15, BMP-16, BMP-17, BMP-18, and MP52/GDF-5;
and
(c) allowing the at least one active agent to promote hair growth.
2. A method of treating a hair loss disorder comprising:
(a) identifying a patient suffering from a hair loss disorder;
(b) administering a composition comprising a therapeutically
effective amount of at least one BMP chosen from the group consisting of BMP-2,
BMP-3,. BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-.
12, BMP-13, BMP-14, BMP-15, BMP-16, BMP-17, BMP-18, and MP52/GDF-5;
(c) administering a therapeutically effective amount of at least
one compound chosen from the group consisting of prednisone, dexamethisone,
hydrocortisone, cyclosporin A, pimecroiimus, tacrolimus, minoxidil, and
finasteride; and
(d) allowing the active agents to promote hair growth.

3. The method of claim 1, wherein the hair loss disorder is chosen from
the group comprising: alopecia areata, alopecia totalis, alopecia universalis,
androgemc alopecia, telogen effluvium, anagen effluvium, and chemotherapy-
induced alopecia.
4. The method of claim 3, wherein the hair loss disorder is alopecia
areata, alopecia totalis, or alopecia universalis.

5. The method of claim 1, wherein the composition comprises a
therapeutically effective amount of BMP-2.
6. The method of claim 1, wherein the composition comprises a
therapeutically effective amount of BMP-4.
. 7. The method ofclaim 1, wherein the composition comprises a
therapeutically effective amount of BMP-2 and a therapeutically effective amount
of BMP-4.
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WO 2006/105359 PCT/US2006/011799
8. The method of claim 1 /wherein the subject is a mammal.
9. The method of claim 1, wherein the subject is a human.
10. The' method of claim 1, wherein the composition is administered by
injection, intraosseously, intravenously, parenterally. percutaneously or extra-
corporeally.
11. The method of claim 1, wherein the therapeutically effective amount
of at least one BMP is chosen from about 0.05 to about 500 mg, from about 0.5 to
about 50 mg, from about 1 to about 25 mg, and from about 5 to about 10 mg.
12. The method of ciaim 1, wherein the therapeutically effective amount
of at least one BMP is administered at a concentration chosen from about 0.001
mg/ml to about 100 mg/ml, from about 0.01 mg/ml to about 50 mg/ml, from about
0.1 mg/ml to about 25 mg/ml, and from about 1 mg/m! to about 5 mg/rni:
13. The method of claim 1, wherein the therapeutically effective amount
of at least one BMP is administered to the subject at intervals chosen from: daily,
weekly, monthly, bi-monthly, tri-monthly, bi-annually, and annually.
14. The method of claim 1, wherein the therapeuticaily effective amount
of at least one BMP is administered to the subject for a period of time chosen
from: about 1 to about 4 weeks, about 5 to about 24 weeks, about 25 to about 52
weeks, about 1 to about 2 years, about 2 to about 5 years, about 5 to about 10
years and about 10 to about 20 years.
15. A method of treating an autoimmune disorder comprising:

(a) identifying a patient suffering from an autoimmune disorder;
(b) administering a composition comprising a therapeutically
effective amount of at least one BMP chosen from the group consisting of BMP-2,
BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-
12, BMP-13, BMP-14, BMP-15, BMP-16, BMP-17, BMP-18, and MP52/GDF-5;
and
(c) allowing the at least one BMP to inhibit immune system .
activity.
16.. A method of treating an autoimmune disorder comprising:
(a) identifying a patient suffering from an autoimmune disorder;
(b) administering a composition comprising a therapeutically
effective amount of at least one BMP chosen from the group consisting of BMP-2,
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WO 2006/105359 PCT/US2006/011799
BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-
12, BMP-13, BMP-14, BMP-15, BMP-16, BMP-17, BMP-18, and MP52/GDF-5;
(c) administering a therapeutically effective amount of at least
one compound chosen, from the group consisting of cyclosporin A, pimecrolimus,
tacrolimus, azathioprine, mycophenolate mofetil, rapamycin, CCl-779,
methotrexate, leflunomide, interferon-(3, copaxone, budenoside, epidermal growth
factor, sulfasalazine, 6-mercaptopurine, azathioprine, metronidazole, mesalamine,
olsalazine, ciprofioxacin, and lignocaine; and
(d) allowing the BMPs to inhibit immune system activity..
17. The method of claim 15 or claim 16, wherein the autoimmune
disorder is chosen from the group comprising: ankylosing spondylitis,
antiphospholipid syndrome, Addison's Disease, autoimmune hemolytic anemia,
autoimmune hepatitis, autoimmune lymphoproliferative syndrome (ALPS),
autoimmune thrombocytopenic purpura (ATP), Behcet's Disease, bulbous
pemphigoid, cardiomyopathy, celiac disease, Dermatitis Herpetiformis, chronic
fatigue immune dysfunction syndrome (CFIDS), chronic inflammatory
demyelinating polyneuropathy (CIDP), cicatricial pemphigoid, systemic sclerosis
(CREST Syndrome), cold aggiutinin disease, Crohn's Disease, cutaneous
vasculitis, Degos1 Disease, dermatomyositis, juvenile dermatomyositis, discoid
lupus erythematosus, essential mixed cryoglobulinemia, fibromyalgia,
Goodpasture's Syndrome, Graves' Disease, Guillain-Barre Syndrome,
Hashimoto's Thyroiditis, idiopathic pulmonary fibrosis, idiopathic
thrombocytopenia purpura (ITP), immunoglobulin A nephropathy, insulin-
dependent diabetes mellitus, juvenile arthritis, Kawasaki's Disease, lichen planus,
membranous glomerulonephritis, Meniere's Disease, mixed connective tissue
disease, multifocal motor neuropathy, multiple sclerosis, myasthenia gravis,
pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polychondritis,
polyglandular syndrome, polymyalgia rheumatica, polymyositis and
dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis,
psoriasis, Raynaud's Phenomenon, Reiter's Syndrome, rheumatic fever,
rheumatoid arthritis, sarcoidosis, scleroderma, Sjogren's Syndrome, Stiff-Man
Syndrome, systemic lupus erthryomatosus, Takayasu arteritis, temporal
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arteritis/giant cell arteritis, ulcerative colitis, uveitis, vasculitis, vitiligo, and
Wegener's Granulomatosis.
18. The method of claim 17, wherein the composition comprises a
therapeutically effective amount of BMP-2,
19. The method of claim 17, wherein the composition comprises a
therapeutically effective amount of BMP-4.
20. The method of claim 17, wherein the composition comprises a
therapeutically effective amount of BMP-2 and a therapeutically effective amount
of BMP-4.
21. The method of claim 17, wherein the subject is a mammal.
22. The method of claim 17, wherein the subject is a human.
23. The method of claim 17, wherein the composition is administered by
injection, intraosseously, intravenously, parenterally, percutaneously or extra-
corporeally.
24.. The method of claim 17, wherein the therapeutically-effective
amount of at least one BMP is chosen from about 0.05 to about 500 mg, from
about 0.5 to about 50 mg, from about 1 to about 25 mg, and from about 5 to about
10 mg..
25. The method of claim 17, wherein the therapeutically effective
amount of at least one BMP is administered at a concentration chosen from about
0.001 mg/ml to about 100 mg/ml, from about 0.01 mg/ml to about 50 mg/ml, from
about 0.1 mg/ml to about 25 mg/ml, and from about 1 mg/ml to about 5 mg/ml.
26. The method of claim 17, wherein the therapeutically effective
amount of at least one BMP is administered to the subject at intervals chosen
from: daily, weekly, monthly, bi-monthly, tri-monthly, bi-annually, and annually.
27. The method of claim 17, wherein the therapeutically effective
amount of at least one BMP is administered to the subject for a period of time
chosen from: about 1 to about 4 weeks, about 5 to about 24 weeks, about 25 to
about 52 weeks, about 1 to about 2 years, about 2 to about 5 years, about 5 to
about 10 years and about 10 to about 20 years.
28. The method of claim 18, wherein the autoimmune disorder is
Crohn's disease, inflammatory bowel disease, multiple sclerosis, psoriasis,
rheumatoid arthritis, or systemic lupus erythematosus.
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WO 2006/105359 PCT/US2006/011799
29. Use of a composition comprising a therapeutically effective amount
of at least one BMP chosen from the group consisting of BMP-2, BMP-3, BMP-4,
BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13,
BMP-14, BMP-15, BMP-16, BMP-17, BMP-18, and MP52/GDF-5 for manufacture
of a medicament for promoting hair growth in a patient suffering from a hair loss
disorder.
30. Use of a composition comprising:

(a) a therapeutically effective amount of at least one BMP chosen
from the group consisting of BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7,
BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13, BMP-14, BMP-15, BMP-16,
BMP-17, BMP-18, and MP52/GDF-5; and
(b) a therapeutically effective amount of at least one compound
chosen from the group consisting of prednisone, dexamethisone, hydrocortisone,
cyclosporin A, pimecrolimus, tacrolimus, minoxidil, and finasteride,
for manufacture of a medicament for promoting hair growth in a patient
suffering from a hair loss disorder.
31. The use of claim 29 or claim 30, wherein the hair loss disorder is
chosen from the group consisting of alopecia areata, alopecia totalis, alopecia
universalis, androgenic alopecia, telogen effluvium, anagen effluvium, and
chemotherapy-induced alopecia.
32. The use of claim 31, wherein the hair loss disorder is alopecia
areata, alopecia totalis, or alopecia universalis.
33. The use according to any one of claims 29 to 32, wherein the
composition comprises a therapeutrcally effective amount of BMP-2.
34. The use according to any one of claims 29 to 32, wherein the
composition comprises a therapeutically effective amount of BMP-4.
35. The use according to any one of claims 29 to 32, wherein the-
composition comprises a therapeutically effective amount of BMP-2 and a
therapeutically effective amount of BMP-4.
36. The use according to any one of claims 29 to 35, wherein the patient
is a mammal. .
37. . The use according to any one of claims 29 to 36, wherein the "patient
is a human.
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WO 2006/105359 PCT/US2006/011799
38. The use according to any one of claims 29 to 37, wherein the
composition is administered by injection, intraosseously, intravenously,
parenterally, percutaneously or extra-corporeally.
39. The use according to any one of claims 28 to 38 wherein, the
therapeutically effective amount of at least one BMP is chosen from about 0.05 to
about 500 mg, from about 0.5 to about 50 mg, from about 1 to about 25 mg, and
from about 5 to about 10 mg.
40. The use according to any one of claims 29 to 38, wherein the
therapeutically effective amount of at least one BMP is administered at a
concentration chosen from about 0.001 mg/mi to about 100 mg/ml, from about
0.01 mg/ml to about 50 mg/ml, from about 0.1 mg/ml to about 25 mg/ml, and from
about 1 mg/ml to about 5 mg/mlP .
41. The use according to any one of claims 29 to 40, wherein the
therapeutically effective amount of at least one BMP is administered to the patient
at intervals chosen from: daily, Weekly, monthly, bi-monthly, tri-monthly, bi-
annually, and annually.
42. The use according to any one of claims 29 to 41, wherein the
therapeutically effective amount of at least one BMP is administered to the patient
for a period of time chosen from: about 1 to about 4 weeks, about 5 to about 24
weeks, about 25 to about 52 weeks, about 1 to about 2 years, about 2 to about 5
years, about 5 to about 10 years and about 10 to about 20 years.

43. Use of a composition comprising a therapeutically effective amount
of at least one BMP chosen from the group consisting of BMP-2, BMP-3, BMP-4,
BMP-5, BMP-6, BMP-7, BMP-a,.BMP-9, BMP-10, BMP-11, BMP-12, BMP-13,
BMP-14.BMP-15, BMP-16, BMP-17, BMP-18, and MP52/GDF-5 for manufacture
of a medicament for inhibiting Immune system activity in a patient suffering from
an autoimmune disorder.
44. Use of a composition comprising:
(a) a therapeutically effective amount of at least one BMP chosen
from the group consisting of BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7,
BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13, BMP-14, BMP-15, BMP-16,
BMP-17, BMP-18, and MP52/GDF-5; and .
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WO 2006/105359 PCT/US2006/011799
(b) a therapeutically effective amount of at least one compound
chosen from the group consisiting of cyclosporin A, pimecrolimus, tacrolimus,-
azathioprine, mycophenolate mofetil, rapamycin, CCI-779, methotrexate,
lefiunomide, interferon-β, copaxone, budenoside, epiderrnai growth factor,
sulfasalazine, 6-mercaptopurine, azathioprine, metronidazole, mesalamine,
olsalazine, ciproflqxacin, and lignocaine,
for manufacture of a medicament for inhibiting immune system activity in a
patient suffering from an autoimmune disorder.
45. . The use of claim 43 or claim 44, wherein the autoimmune disorder is
chosen from the group consisting of ankylosing spondylitis, antiphosphoiipid
syndrome, Addison's Disease, autoimmune hemolytic anemia, autoimmune
hepatitis, autoimmune lymphoproliferative syndrome (ALPS), autoimmune
thrombocytopenic purpura (ATP), Behcet's Disease, bulbous pemphigoid,
cardiomyopathy, celiac disease, Dermatitis Herpetiformis, chronic fatigue immune
dysfunction syndrome (CFIDS), chronic inflammatory demyelinating
polyneuropathy (CIDP), cicatricial pemphigoid, systemic sclerosis (CREST
Syndrome), cold agglutinin disease, Crohn's Disease, cutaneous vasculitis,
Degos' Disease, dermatomyositis, juvenile dermatomyositis, discoid lupus-
erythernatosus, essential mixed cryoglobulinemia, fibromyalgia, Goodpasture's
Syndrome, Graves' Disease, Guillain-Barre Syndrome, Hashimoto's Thyroiditis,
idiopathic pulmonary fibrosis, idiopathic thrombocytopenia purpura (ITP),
immunoglobulin A nephropathy, insulin-dependent diabetes mellitus, juvenile
arthritis, Kawasaki's Disease, lichen planus, membranous glomerulonephritis,,
Meniere's Disease, mixed connective tissue disease, multifocal motor neuropathy,
multiple sclerosis, myasthenia gravis, pemphigus vulgaris, pernicious anemia,
polyarteritis nodosa, polychondritis, polyglandular syndrome, polymyalgia
rheumatica, polymyositis and dermatomyositis, primary agammaglobulinemia,
primary biliary cirrhosis, psoriasis, Raynaud's Phenomenon, Reiter's Syndrome,
rheumatic fever, rheumatoid arthritis, sarcoidosis, scleroderma, Sjogren's
Syndrome, Stiff-Man Syndrome, systemic lupus erthryomatosus, Takayasu
arteritis, temporal arteritis/giant cell arteritis, ulcerative colitis, uveitis, vasculitis,
vitiligp, and Wegener's Granulomatosis.
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WO 2006/105359 PCT/US2006/011799
46. The use according to any one of claims 43 to 45, wherein the
composition comprises a therapeutically effective amount of BMP-2.
47. The use according to any one of claims 43 to 45, wherein the
composition comprises a therapeutical!'/ effective amount of BMP-4..
48. The use according to any one of claims 43 to 45, wherein the
composition comprises a. therapeutically effective amount of BMP-2 and a
therapeutically effective amount of BMP-4.
49. The use according to any one of claims 43 to 48, wherein the
subject is a mammal.
50. The use according to any one of claims 43 to 49, wherein the
subject is a human.
51. The use according to any one of. claims 43 to 50, wherein the
composition is administered by injection, intraosseously, intravenously,
parenterally, percutaneously or extra-corporeally.
52. The use according to any one of claims 43 to 51, wherein the
therapeutically effective amount of at least one BMP is chosen from about 0.05 to
about 500 mg, from about 0.5 to about 50 mg, from about 1 to about 25 mg, and
from about 5 to about 10 mg..
53. The use according to any one of claims 43 to 51, wherein the
therapeutically-effective amount.of at least one BMP is administered at a
concentration chosen from about 0.001 mg/ml to about 100 mg/ml, from about
0.01 mg/ml to about 50 mg/ml, from about 0.1 mg/ml to about 25 mg/ml, and from,
about 1 mg/ml to about 5 mg/ml.
54. The use according to any one of claims 43 to 53, wherein the
therapeutically effective amount of at least one BMP is administered to the subject
at intervals chosen from: daily, weekly, monthly, bi-monthly, tri-monthly, bi-
annually, and annually. • '
55. The use according to any one of claims 43 to 54, wherein the
therapeutically effective amount of at least one BMP is administered to the subject
for a period of time chosen from: about 1 to about 4 weeks, about 5. to about 24
weeks, about 25 to about 52 weeks, about 1 to about 2 years, about 2 to about 5
years, about 5 to about 10 years and about 10 to about 20 years.
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WO 2006/105359 PCT/US2006/011799
56. The use according to any one of claims 43 to 55, wherein the
autoimmune disorder is Crohn's disease, inflammatory bowel disease, multiple
sclerosis, psoriasis, rheumatoid arthritis, or systemic lupus erythematosus.
57. Use of a composition comprising a therapeutically effective amount
of at least one BMP chosen from the group consisting of BMP-2, BMP-3, BMP-4,
BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13,
BMP-14, BMP-15, BMP-16, BMP-17, BMP-18, and MP52/GDF-5 for manufacture
of a medicament for promoting hair growth.
58. A pharmaceutical composition comprising:

(a) at least one BMP chosen from the group consisting of BMP-2,
BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-
12, BMP-13, BMP-14, BMP-15, BMP-16, BMP-17, BMP-18, and MP52/GDF-5;
(b) a sterile solution of 0.5% sucrose, 2.5% glycine, 5 mM L-
glutamic acid, 5 mM NaCI, and 0.01% polysorbate 80, at pH 4.50.
59. A pharmaceutical composition for promoting hair growth comprising:
(a) at least one BMP chosen from the group consisting of BMP-
2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11,
BMP-12, BMP-13, BMP-14, BMP-15, BMP-16, BMP-17, BMP-18, and '
MP52/GDF-5; and
(b) at least one compound chosen from the group consisting of
prednisone, dexamethisone, hydrocortisone, cyclosporin A, pimecrolimus,
tacrolimus, minoxidil, and finasteride.
60. A pharmaceutical composition for inhibiting immune system activity
comprising at least one BMP chosen from the group consisting of BMP-2, BMP-3,
BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9, BMP-10, BMP-11, BMP-12,.
BMP-13, BMP-14.BMP-15, BMP-16, BMP-17, BMP-18, and MP52/GDF-5, and at
least one compound chosen from the group consisting of cyclosporin A,
pimecrolimus, tacrolimus, azathioprine, mycophenolate mofetil, rapamycin, CCI-
779, methotrexate, leflunomide, interferon-(3, copaxone, budenoside, epidermal
growth factor, sulfasalazine, 6-mercaptopurine, azathioprine, metronidazole,
mesalamine, olsalazine, ciprofloxacin, and lignocaine.
43

Methods and compositions for stimulating
hair growth and inhibiting immune system activity by
administering BMPs are provided. The methods and
compositions can be used for treating or preventing
disorders resulting in loss of hair, as well as a wide range of
autoimmune disorders.