U.S. patent application number 17/390096 was filed with the patent office on 2022-01-20 for ascaroside treatment of autoimmune and inflammatory diseases.
The applicant listed for this patent is Boyce Thompson Institute for Plant Research, California Institute of Technology. Invention is credited to Andrea Choe, Frank C, Schroeder, Paul W. Sternberg.
Application Number | 20220016182 17/390096 |
Document ID | / |
Family ID | |
Filed Date | 2022-01-20 |
United States Patent
Application |
20220016182 |
Kind Code |
A1 |
Choe; Andrea ; et
al. |
January 20, 2022 |
ASCAROSIDE TREATMENT OF AUTOIMMUNE AND INFLAMMATORY DISEASES
Abstract
The present invention relates to the use of ascr#7 for
preventing or treating IL-6 and/or IL-1.beta.-mediated
diseases.
Inventors: |
Choe; Andrea; (Pasadena,
CA) ; Sternberg; Paul W.; (San Marino, CA) ;
Schroeder; Frank C,; (Ithaca, NY) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
California Institute of Technology
Boyce Thompson Institute for Plant Research |
Pasadena
Ithaca |
CA
NY |
US
US |
|
|
Appl. No.: |
17/390096 |
Filed: |
July 30, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16327659 |
Feb 22, 2019 |
11077151 |
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PCT/US17/48665 |
Aug 25, 2017 |
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17390096 |
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62379651 |
Aug 25, 2016 |
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International
Class: |
A61K 35/62 20060101
A61K035/62; A61P 1/04 20060101 A61P001/04; A61K 31/4178 20060101
A61K031/4178; A61K 31/7028 20060101 A61K031/7028; A61K 45/06
20060101 A61K045/06; A61P 37/06 20060101 A61P037/06; A61P 3/10
20060101 A61P003/10; A61K 31/573 20060101 A61K031/573; G01N 33/68
20060101 G01N033/68; A61K 31/4439 20060101 A61K031/4439; A61K 31/56
20060101 A61K031/56; A61K 31/7042 20060101 A61K031/7042; A61K 47/26
20060101 A61K047/26 |
Claims
1.-16. (canceled)
17. A composition comprising ascr#7 and a corticosteroid.
18. The composition of claim 17, wherein the corticosteroid is
selected from aldosterone, betamethasone, budesonide,
corticosterone, cortisol, cortisone, dexamethasone, fluticasone,
hydrocortisone, methylprednisolone, prednisolone, and
prednisone.
19. The composition of claim 17, wherein ascr#7 and a
corticosteroid are administered in the same formulation.
20. The composition of claim 17, further comprising a
pharmaceutically acceptable carrier.
21. The composition of claim 17, wherein the composition has
synergistic effects of ascr#7 and a corticosteroid.
22. The composition of claim 17, wherein ascr#7 is in a
pharmaceutically acceptable salt or prodrug form.
23. The composition of claim 17, wherein the composition can be
used for preventing or treating an IL-6- and/or IL-1.beta.-mediated
disease in a subject.
24. The composition of claim 23, wherein the IL-6- and/or
IL-1.beta.-mediated disease is an autoimmune disease or an
inflammatory disease.
25. The composition of claim 23, wherein the subject has an
elevated level of IL-6 and/or IL-1.beta..
26. The composition of claim 23, wherein the level of IL-6 and/or
IL-1.beta. in an affected tissue of the subject exceeds 200% of a
normal level in the affected tissue.
27. The composition of claim 23, wherein the disease is
Agammaglobulinemia, Amyloidosis, Ankylosing spondylitis,
Anti-GBM/Anti-TBM nephritis, Antiphospholipid syndrome, Autoimmune
hepatitis, Autoimmune inner ear disease, Atopic dermatitis, Asthma,
Castleman disease, Celiac disease, Chagas disease, Chronic
recurrent multifocal osteomyelitis, Cogan's syndrome, Cold
agglutinin disease, CREST syndrome, Crohn's disease,
Dermatomyositis, Devic's disease (neuromyelitis optica), Discoid
lupus, Endometriosis, Eosinophilic esophagitis, Eosinophilic
fasciitis, Evan's syndrome, Fibromyalgia, Giant cell arteritis,
Giant cell myocarditis, Glomerulonephritis, Goodpasture's syndrome,
Granulomatosis with polyangiitis, Graves' disease, Guillain-Barre
syndrome, Hashimoto's thyroiditis, Hemolytic anemia,
Henoch-Schonlein purpura, Hypogammaglobulinemia, Hypoproliferative
anemia, IgA Nephropathy, Inclusion body myositis, Interstitial
cystitis, Inflammatory Bowel Disease, Juvenile arthritis,
Juvenile/Type 1 Diabetes, Juvenile myositis, Kawasaki syndrome,
Lichen planus, Lichen sclerosus, Lupus (SLE), Meniere's disease,
Multiple sclerosis, Myasthenia gravis, Microscopic polyangiitis,
Optic neuritis, Pemphigus, Polyarteritis nodosa, Polymyalgia
rheumatica, Polymyositis, Primary biliary cirrhosis, Primary
sclerosing cholangitis, Psoriasis, Psoriatic arthritis, Rheumatic
fever, Rheumatoid arthritis, Sarcoidosis, Sjogren's syndrome,
Temporal arteritis/Giant cell arteritis, Transverse myelitis,
Ulcerative colitis, Uveitis, Vasculitis, Vitiligo, Viral
myocarditis, or Wegener's granulomatosis (Granulomatosis with
Polyangiitis (GPA)).
28. The composition of claim 27, wherein the disease is asthma.
29. The composition of claim 27, wherein the disease is
inflammatory bowel disease.
30. The composition of claim 27, wherein the disease is type 1
diabetes.
31. The composition of claim 27, wherein the disease is not
eosinophilic esophagitis (EoE).
32. The composition of claim 23, wherein the subject is a
mammal.
33. The composition of claim 32, wherein the mammal is a human.
34. The composition of claim 17, wherein the composition reduces
levels of IL-6 and/or IL-1.beta..
35. The composition of claim 17, wherein the composition reduces
disease progression of autoimmune and/or inflammatory disease.
36. The composition of claim 35, wherein the autoimmune and/or
inflammatory disease is asthma, type 1 diabetes, or inflammatory
bowel disease.
Description
RELATED APPLICATIONS
[0001] This application claims the benefit of and priority to U.S.
Provisional Patent Application Ser. No. 62/379,651, filed Aug. 25,
2016, hereby incorporated by reference in its entirety.
BACKGROUND
[0002] Many nematode species are known to parasitize humans, a
relationship that is thought to have existed for thousands of
years.
[0003] It has long been known that there is an inverse relationship
between areas endemic with parasitic nematodes and areas with a
high incidence of autoimmune disease. This observation spurred the
hypothesis that parasitic nematodes may protect against autoimmune
disease, and it has been corroborated by numerous human clinical
trials and animal model studies, where ingestion of parasitic
nematodes has alleviated disease symptoms.
[0004] Some nematode-based approaches for treating autoimmune and
inflammatory diseases have been identified. A need exists for
additional disease treatments for which factors and/or isolates of
parasitic nematodes may offer possibilities.
SUMMARY
[0005] The present invention is based, at least in part, on the
discovery that ascarosides, a family of small-molecule nematode
pheromones, modulate the immune response and have a therapeutic
effect on various autoimmune and inflammatory disorders. One of the
ascarosides, ascr#7, markedly inhibits the hallmark pathologies in
mouse models of asthma, inflammatory bowel disease, and type 1
diabetes, and significantly reduces both IL-6 and IL-1.beta.
secretion. These findings suggest that ascr#7 has a therapeutic
effect in diseases where elevated levels of IL-6 and/or IL-1.beta.
contribute to the diseases' pathogeneses.
[0006] In some aspects, the invention provides a method of
preventing, alleviating, or treating an IL-6- and/or
IL-1.beta.-mediated disease in a subject. The method comprises
administering to the subject a composition comprising ascr#7 or a
pharmaceutically acceptable salt or prodrug thereof.
[0007] Numerous embodiments are described that can be applied to
any aspect of the present invention and/or combined with any other
embodiment described herein. For example, in some embodiments, the
subject has an elevated level of IL-6 and/or IL-1.beta.. In some
other embodiments, IL-6 and/or IL-1.beta. contribute to one or more
symptoms of the disease. In such embodiments, the method may
comprise assessing the level of IL-6 and/or IL-1.beta. in an
affected tissue of the subject and, if the level exceeds 200% of a
normal level for the affected tissue, administering ascr#7 or a
pharmaceutically acceptable salt or prodrug thereof.
[0008] In some embodiments, the disease is Agammaglobulinemia,
Amyloidosis, Ankylosing spondylitis, Anti-GBM/Anti-TBM nephritis,
Antiphospholipid syndrome, Autoimmune hepatitis, Autoimmune inner
ear disease, Atopic dermatitis, Asthma, Castleman disease, Celiac
disease, Chagas disease, Chronic recurrent multifocal
osteomyelitis, Cogan's syndrome, Cold agglutinin disease, CREST
syndrome, Crohn's disease, Dermatomyositis, Devic's disease
(neuromyelitis optica), Discoid lupus, Endometriosis, Eosinophilic
esophagitis, Eosinophilic fasciitis, Evan's syndrome, Fibromyalgia,
Giant cell arteritis, Giant cell myocarditis, Glomerulonephritis,
Goodpasture's syndrome, Granulomatosis with polyangiitis, Graves'
disease, Guillain-Barre syndrome, Hashimoto's thyroiditis,
Hemolytic anemia, Henoch-Schonlein purpura, Hypogammaglobulinemia,
Hypoproliferative anemia, IgA Nephropathy, Inclusion body myositis,
Interstitial cystitis, Inflammatory Bowel Disease, Juvenile
arthritis, Juvenile/Type 1 Diabetes, Juvenile myositis, Kawasaki
syndrome, Lichen planus, Lichen sclerosus, Lupus (SLE), Meniere's
disease, Multiple sclerosis, Myasthenia gravis, Microscopic
polyangiitis, Optic neuritis, Pemphigus, Polyarteritis nodosa,
Polymyalgia rheumatica, Polymyositis, Primary biliary cirrhosis,
Primary sclerosing cholangitis, Psoriasis, Psoriatic arthritis,
Rheumatic fever, Rheumatoid arthritis, Sarcoidosis, Sjogren's
syndrome, Temporal arteritis/Giant cell arteritis, Transverse
myelitis, Ulcerative colitis, Uveitis, Vasculitis, Vitiligo, Viral
myocarditis, or Wegener's granulomatosis (Granulomatosis with
Polyangiitis (GPA)).
[0009] In certain preferred embodiments, the disease is asthma,
inflammatory bowel disease, or type 1 diabetes. In some preferred
embodiments, the disease is eosinophilic esophagitis (EoE), while
in other embodiments the disease is not EoE.
[0010] In some embodiments, the method further comprises
administering an agent with an anti-inflammatory effect, such as a
corticosteroid.
[0011] In some embodiments, the method further comprises
administering a corticosteroid to the subject. In some embodiments,
the corticosteroid is selected from aldosterone, betamethasone,
budesonide, corticosterone, cortisol, cortisone, dexamethasone,
fluticasone (e.g., fluticasone propionate), hydrocortisone,
methylprednisolone, prednisolone, and prednisone. In some
embodiments, the subject is a mammal. In certain embodiments, the
mammal is a mouse or a human. In certain preferred embodiments, the
mammal is a human.
[0012] In some aspects, the invention provides a method of reducing
IL-6 and/or IL-1.beta. production from a cell, comprising
contacting the cell with a composition comprising ascr#7 or a
pharmaceutically acceptable salt or prodrug thereof.
BRIEF DESCRIPTION OF THE DRAWINGS
[0013] FIG. 1 shows that ascr#7 diminishes IL-6 production by
testing dendritic cells in an in vitro assay.
[0014] FIG. 2 shows that ascr#7 diminishes production by testing
dendritic cells in an in vitro assay.
[0015] FIG. 3A shows a diagram of a mouse in a respiratory
chamber.
[0016] FIG. 3B shows that ascr#7 prevents hallmark pathologies of
asthma. FIG. 3B shows that ascr#7 diminishes resistance to air flow
in a mouse model of asthma.
[0017] FIG. 3C shows that ascr#7 decreases the inflammatory
buildup, airway restriction, and mucus secretion based on the
staining of the histological sections of lungs.
[0018] FIG. 4A shows a diagram of a mouse having a blood sample
taken from the tail vein.
[0019] FIG. 4B shows that ascr#7 delays onset of type 1 diabetes in
Non-obese diabetic (NOD) mice.
[0020] FIG. 4C shows that ascr#7 delays onset of type 1 diabetes in
Non-obese diabetic (NOD) mice.
[0021] FIG. 5 shows that ascr#7 markedly reduces colon shortening
and diarrhea in an immune dysregulation model of Inflammatory Bowel
Disease (IBD). The inset shows a diagram of the colon.
DETAILED DESCRIPTION
[0022] It has been determined herein that ascr#7 has a therapeutic
effect in autoimmune and/or inflammatory diseases, e.g., where
elevated levels of IL-6 and/or IL-1.beta. contribute to the
diseases' pathogeneses. Accordingly, the present invention relates,
in part, to methods of alleviating, treating, or preventing an
IL-6- and/or IL-1.beta.-mediated disease in a subject by
administering to the subject a composition comprising ascr#7 or a
pharmaceutically acceptable salt or prodrug thereof. In other
aspects, the present invention relates to methods of reducing IL-6
and/or IL-1.beta. production from a cell by contacting the cell, or
affiliated cells, with a composition comprising ascr#7 or a
pharmaceutically acceptable salt or prodrug thereof.
I. Definitions
[0023] For convenience, certain terms employed in the
specification, and appended claims are collected here.
[0024] The term "comprise" or variations such as "comprises" or
"comprising" will be understood to imply the inclusion of a stated
integer (or components) or group of integers (or components), but
not the exclusion of any other integer (or components) or group of
integers (or components).
[0025] The singular forms "a," "an," and "the" include the plurals
unless the context clearly dictates otherwise.
[0026] The term "including" is used to mean "including but not
limited to." "Including" and "including but not limited to" are
used interchangeably.
[0027] The terms "subject" refer to either a human or a non-human
animal. This term includes mammals such as humans, primates,
livestock animals (e.g., bovines, porcines), companion animals
(e.g., canines, felines) and rodents (e.g., mice, rabbits and
rats).
[0028] "About" and "approximately" shall generally mean an
acceptable degree of error for the quantity measured given the
nature or precision of the measurements. Typically, exemplary
degrees of error are within 20%, preferably within 10%, and more
preferably within 5% of a given value or range of values.
Alternatively, and particularly in biological systems, the terms
"about" and "approximately" may mean values that are within an
order of magnitude, preferably within 5-fold and more preferably
within 2-fold of a given value. Numerical quantities given herein
are approximate unless stated otherwise, meaning that the term
"about" or "approximately" can be inferred when not expressly
stated.
[0029] The term "prodrug" is intended to encompass compounds which,
under physiologic conditions, are converted into the
therapeutically active agents of the present invention (e.g.,
ascr#7). A common method for making a prodrug is to include one or
more selected moieties which are hydrolyzed under physiologic
conditions to reveal the desired molecule. In other embodiments,
the prodrug is converted by an enzymatic activity of the subject.
For example, esters or carbonates (e.g., esters or carbonates of
alcohols or carboxylic acids) are preferred prodrugs of the present
invention. In certain embodiments, some or all of the compounds of
the present invention (e.g., ascr#7) in a formulation represented
above can be replaced with the corresponding suitable prodrug,
e.g., wherein a hydroxyl in the parent compound is presented as an
ester or a carbonate or carboxylic acid present in the parent
compound is presented as an ester.
[0030] The term "treating" includes prophylactic and/or therapeutic
treatments. The term "prophylactic or therapeutic" treatment is
art-recognized and includes administration to the host of one or
more of the subject compositions. If it is administered prior to
clinical manifestation of the unwanted condition (e.g., disease or
other unwanted state of the host animal) then the treatment is
prophylactic (i.e., it protects the host against developing the
unwanted condition), whereas if it is administered after
manifestation of the unwanted condition, the treatment is
therapeutic, (i.e., it is intended to diminish, alleviate,
ameliorate, or stabilize the existing unwanted condition or side
effects thereof).
[0031] As used herein, a therapeutic that "prevents" a disorder or
condition refers to a compound that, in a statistical sample,
reduces the occurrence of the disorder or condition in the treated
sample relative to an untreated control sample, or delays the onset
or reduces the severity of one or more symptoms of the disorder or
condition relative to the untreated control sample.
[0032] Unless otherwise defined herein, scientific and technical
terms used in this application shall have the meanings that are
commonly understood by those of ordinary skill in the art.
Generally, nomenclature and techniques relating to chemistry,
molecular biology, cell and cancer biology, immunology,
microbiology, pharmacology, and protein and nucleic acid chemistry,
described herein, are those well-known and commonly used in the
art.
II. Compositions
Ascr#7
[0033] Ascr#7 is an ascaroside produced by a nematode, e.g.,
Nippostrongylus brasiliensis.
[0034] Ascr#7 has the structure:
##STR00001##
[0035] The compositions and methods disclosed herein may use any
suitable form, prodrug, and/or salt of ascr#7 that exhibits the
desired therapeutic effects in subjects.
Pharmaceutical Compositions
[0036] In certain embodiments, the present invention provides
pharmaceutical compositions comprising ascr#7 or a pharmaceutically
acceptable salt or prodrug thereof and a pharmaceutically
acceptable carrier.
[0037] The compositions and methods of the present invention may be
utilized to treat a subject in need thereof. In certain
embodiments, the subject is a mammal such as a human, or a
non-human mammal. When administered to an animal, such as a human,
the composition or the compound is preferably administered as a
pharmaceutical composition comprising, for example, a compound of
the invention and a pharmaceutically acceptable carrier.
Pharmaceutically acceptable carriers are well known in the art and
include, for example, aqueous solutions such as water or
physiologically buffered saline or other solvents or vehicles such
as glycols, glycerol, oils such as olive oil, or injectable organic
esters. In a preferred embodiment, when such pharmaceutical
compositions are for human administration, particularly for
invasive routes of administration (i.e., routes, such as injection
or implantation, that circumvent transport or diffusion through an
epithelial barrier), the aqueous solution is pyrogen-free, or
substantially pyrogen-free. The excipients can be chosen, for
example, to effect delayed release of an agent or to selectively
target one or more cells, tissues or organs. The pharmaceutical
composition can be in dosage unit form such as tablet, capsule
(including sprinkle capsule and gelatin capsule), granule, lyophile
for reconstitution, powder, solution, syrup, suppository, injection
or the like. The composition can also be present in a transdermal
delivery system, e.g., a skin patch. The composition can also be
present in a solution suitable for topical administration, such as
an eye drop.
[0038] A pharmaceutically acceptable carrier can contain
physiologically acceptable agents that act, for example, to
stabilize, increase solubility or to increase the absorption of a
compound such as a compound of the invention. Such physiologically
acceptable agents include, for example, carbohydrates, such as
glucose, sucrose or dextrans, antioxidants, such as ascorbic acid
or glutathione, chelating agents, low molecular weight proteins or
other stabilizers or excipients. The choice of a pharmaceutically
acceptable carrier, including a physiologically acceptable agent,
depends, for example, on the route of administration of the
composition. The preparation or pharmaceutical composition can be a
self-emulsifying drug delivery system or a self-microemulsifying
drug delivery system. The pharmaceutical composition (preparation)
also can be a liposome or other polymer matrix, which can have
incorporated therein, for example, a compound of the invention.
Liposomes, for example, which comprise phospholipids or other
lipids, are nontoxic, physiologically acceptable and metabolizable
carriers that are relatively simple to make and administer.
[0039] The phrase "pharmaceutically acceptable" is employed herein
to refer to those compounds, materials, compositions, and/or dosage
forms which are, within the scope of sound medical judgment,
suitable for use in contact with the tissues of a subject without
excessive toxicity, irritation, allergic response, or other problem
or complication, commensurate with a reasonable benefit/risk
ratio.
[0040] The phrase "pharmaceutically acceptable carrier" as used
herein means a pharmaceutically acceptable material, composition or
vehicle, such as a liquid or solid filler, diluent, excipient,
solvent or encapsulating material. Each carrier must be
"acceptable" in the sense of being compatible with the other
ingredients of the formulation and not injurious to the subject.
Some examples of materials which can serve as pharmaceutically
acceptable carriers include: (1) sugars, such as lactose, glucose
and sucrose; (2) starches, such as corn starch and potato starch;
(3) cellulose, and its derivatives, such as sodium carboxymethyl
cellulose, ethyl cellulose and cellulose acetate; (4) powdered
tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such
as cocoa butter and suppository waxes; (9) oils, such as peanut
oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil
and soybean oil; (10) glycols, such as propylene glycol; (11)
polyols, such as glycerin, sorbitol, mannitol and polyethylene
glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13)
agar; (14) buffering agents, such as magnesium hydroxide and
aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water;
(17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol;
(20) phosphate buffer solutions; and (21) other non-toxic
compatible substances employed in pharmaceutical formulations.
[0041] The formulations may conveniently be presented in unit
dosage form and may be prepared by any methods well known in the
art of pharmacy. The amount of active ingredient which can be
combined with a carrier material to produce a single dosage form
will vary depending upon the subject being treated, the particular
mode of administration. The amount of active ingredient that can be
combined with a carrier material to produce a single dosage form
will generally be that amount of the compound which produces a
therapeutic effect. Generally, out of one hundred percent, this
amount will range from about 1 percent to about ninety-nine percent
of active ingredient, preferably from about 5 percent to about 70
percent, most preferably from about 10 percent to about 30
percent.
[0042] Methods of preparing these formulations or compositions
include the step of bringing into association an active compound,
such as a compound of the invention, with the carrier and,
optionally, one or more accessory ingredients. In general, the
formulations are prepared by uniformly and intimately bringing into
association a compound of the present invention with liquid
carriers, or finely divided solid carriers, or both, and then, if
necessary, shaping the product.
[0043] Formulations of the invention suitable for oral
administration may be in the form of capsules (including sprinkle
capsules and gelatin capsules), cachets, pills, tablets, lozenges
(using a flavored basis, usually sucrose and acacia or tragacanth),
lyophile, powders, granules, or as a solution or a suspension in an
aqueous or non-aqueous liquid, or as an oil-in-water or
water-in-oil liquid emulsion, or as an elixir or syrup, or as
pastilles (using an inert base, such as gelatin and glycerin, or
sucrose and acacia) and/or as mouth washes and the like, each
containing a predetermined amount of a compound of the present
invention as an active ingredient. Compositions or compounds may
also be administered as a bolus, electuary or paste.
[0044] To prepare solid dosage forms for oral administration
(capsules (including sprinkle capsules and gelatin capsules),
tablets, pills, dragees, powders, granules and the like), the
active ingredient is mixed with one or more pharmaceutically
acceptable carriers, such as sodium citrate or dicalcium phosphate,
and/or any of the following: (1) fillers or extenders, such as
starches, lactose, sucrose, glucose, mannitol, and/or silicic acid;
(2) binders, such as, for example, carboxymethylcellulose,
alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia;
(3) humectants, such as glycerol; (4) disintegrating agents, such
as agar-agar, calcium carbonate, potato or tapioca starch, alginic
acid, certain silicates, and sodium carbonate; (5) solution
retarding agents, such as paraffin; (6) absorption accelerators,
such as quaternary ammonium compounds; (7) wetting agents, such as,
for example, cetyl alcohol and glycerol monostearate; (8)
absorbents, such as kaolin and bentonite clay; (9) lubricants, such
a talc, calcium stearate, magnesium stearate, solid polyethylene
glycols, sodium lauryl sulfate, and mixtures thereof; (10)
complexing agents, such as, modified and unmodified cyclodextrins;
and (11) coloring agents. In the case of capsules (including
sprinkle capsules and gelatin capsules), tablets and pills, the
pharmaceutical compositions may also comprise buffering agents.
Solid compositions of a similar type may also be employed as
fillers in soft and hard-filled gelatin capsules using such
excipients as lactose or milk sugars, as well as high molecular
weight polyethylene glycols and the like.
[0045] A tablet may be made by compression or molding, optionally
with one or more accessory ingredients. Compressed tablets may be
prepared using binder (for example, gelatin or hydroxypropylmethyl
cellulose), lubricant, inert diluent, preservative, disintegrant
(for example, sodium starch glycolate or cross-linked sodium
carboxymethyl cellulose), surface-active or dispersing agent.
Molded tablets may be made by molding in a suitable machine a
mixture of the powdered compound moistened with an inert liquid
diluent.
[0046] The tablets, and other solid dosage forms of the
pharmaceutical compositions, such as dragees, capsules (including
sprinkle capsules and gelatin capsules), pills and granules, may
optionally be scored or prepared with coatings and shells, such as
enteric coatings and other coatings well known in the
pharmaceutical-formulating art. They may also be formulated so as
to provide slow or controlled release of the active ingredient
therein using, for example, hydroxypropylmethyl cellulose in
varying proportions to provide the desired release profile, other
polymer matrices, liposomes and/or microspheres. They may be
sterilized by, for example, filtration through a bacteria-retaining
filter, or by incorporating sterilizing agents in the form of
sterile solid compositions that can be dissolved in sterile water,
or some other sterile injectable medium immediately before use.
These compositions may also optionally contain opacifying agents
and may be of a composition that they release the active
ingredient(s) only, or preferentially, in a certain portion of the
gastrointestinal tract, optionally, in a delayed manner. Examples
of embedding compositions that can be used include polymeric
substances and waxes. The active ingredient can also be in
micro-encapsulated form, if appropriate, with one or more of the
above-described excipients.
[0047] Liquid dosage forms useful for oral administration include
pharmaceutically acceptable emulsions, lyophiles for
reconstitution, microemulsions, solutions, suspensions, syrups and
elixirs. In addition to the active ingredient, the liquid dosage
forms may contain inert diluents commonly used in the art, such as,
for example, water or other solvents, cyclodextrins and derivatives
thereof, solubilizing agents and emulsifiers, such as ethyl
alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl
alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol,
oils (in particular, cottonseed, groundnut, corn, germ, olive,
castor and sesame oils), glycerol, tetrahydrofuryl alcohol,
polyethylene glycols and fatty acid esters of sorbitan, and
mixtures thereof.
[0048] Besides inert diluents, the oral compositions can also
include adjuvants such as wetting agents, emulsifying and
suspending agents, sweetening, flavoring, coloring, perfuming and
preservative agents.
[0049] Suspensions, in addition to the active compounds, may
contain suspending agents as, for example, ethoxylated isostearyl
alcohols, polyoxyethylene sorbitol and sorbitan esters,
microcrystalline cellulose, aluminum metahydroxide, bentonite,
agar-agar and tragacanth, and mixtures thereof.
[0050] Formulations of the pharmaceutical compositions for
administration to the mouth may be presented as a mouthwash, or an
oral spray, or an oral ointment.
[0051] Alternatively or additionally, compositions can be
formulated for delivery via a catheter, stent, wire, or other
intraluminal device. Delivery via such devices may be especially
useful for delivery to the bladder, urethra, ureter, rectum, or
intestine.
[0052] Formulations which are suitable for vaginal administration
also include pessaries, tampons, creams, gels, pastes, foams or
spray formulations containing such carriers as are known in the art
to be appropriate.
[0053] Dosage forms for the topical or transdermal administration
include powders, sprays, ointments, pastes, creams, lotions, gels,
solutions, patches and inhalants. The active compound may be mixed
under sterile conditions with a pharmaceutically acceptable
carrier, and with any preservatives, buffers, or propellants that
may be required.
[0054] The ointments, pastes, creams and gels may contain, in
addition to an active compound, excipients, such as animal and
vegetable fats, oils, waxes, paraffins, starch, tragacanth,
cellulose derivatives, polyethylene glycols, silicones, bentonites,
silicic acid, talc and zinc oxide, or mixtures thereof.
[0055] Powders and sprays can contain, in addition to an active
compound, excipients such as lactose, talc, silicic acid, aluminum
hydroxide, calcium silicates and polyamide powder, or mixtures of
these substances. Sprays can additionally contain customary
propellants, such as chlorofluorohydrocarbons and volatile
unsubstituted hydrocarbons, such as butane and propane.
[0056] Transdermal patches have the added advantage of providing
controlled delivery of a compound of the present invention to the
body. Such dosage forms can be made by dissolving or dispersing the
active compound in the proper medium. Absorption enhancers can also
be used to increase the flux of the compound across the skin. The
rate of such flux can be controlled by either providing a rate
controlling membrane or dispersing the compound in a polymer matrix
or gel.
[0057] Ophthalmic formulations, eye ointments, powders, solutions
and the like, are also contemplated as being within the scope of
this invention. Exemplary ophthalmic formulations are described in
U.S. Publication Nos. 2005/0080056, 2005/0059744, 2005/0031697 and
2005/004074 and U.S. Pat. No. 6,583,124, the contents of which are
incorporated herein by reference. If desired, liquid ophthalmic
formulations have properties similar to that of lacrimal fluids,
aqueous humor or vitreous humor or are compatable with such fluids.
A preferred route of administration is local administration (e.g.,
topical administration, such as eye drops, or administration via an
implant).
[0058] The phrases "parenteral administration" and "administered
parenterally" as used herein means modes of administration other
than enteral and topical administration, usually by injection, and
includes, without limitation, intravenous, intramuscular,
intraarterial, intrathecal, intracapsular, intraorbital,
intracardiac, intradermal, intraperitoneal, transtracheal,
subcutaneous, subcuticular, intraarticular, subcapsular,
subarachnoid, intraspinal and intrasternal injection and
infusion.
[0059] Pharmaceutical compositions suitable for parenteral
administration comprise one or more active compounds in combination
with one or more pharmaceutically acceptable sterile isotonic
aqueous or nonaqueous solutions, dispersions, suspensions or
emulsions, or sterile powders which may be reconstituted into
sterile injectable solutions or dispersions just prior to use,
which may contain antioxidants, buffers, bacteriostats, solutes
which render the formulation isotonic with the blood of the
intended recipient or suspending or thickening agents.
[0060] Examples of suitable aqueous and nonaqueous carriers that
may be employed in the pharmaceutical compositions of the invention
include water, ethanol, polyols (such as glycerol, propylene
glycol, polyethylene glycol, and the like), and suitable mixtures
thereof, vegetable oils, such as olive oil, and injectable organic
esters, such as ethyl oleate. Proper fluidity can be maintained,
for example, by the use of coating materials, such as lecithin, by
the maintenance of the required particle size in the case of
dispersions, and by the use of surfactants.
[0061] These compositions may also contain adjuvants such as
preservatives, wetting agents, emulsifying agents and dispersing
agents. Prevention of the action of microorganisms may be ensured
by the inclusion of various antibacterial and antifungal agents,
for example, paraben, chlorobutanol, phenol sorbic acid, and the
like. It may also be desirable to include isotonic agents, such as
sugars, sodium chloride, and the like into the compositions. In
addition, prolonged absorption of the injectable pharmaceutical
form may be brought about by the inclusion of agents that delay
absorption such as aluminum monostearate and gelatin.
[0062] In some cases, in order to prolong the effect of a drug, it
is desirable to slow the absorption of the drug from subcutaneous
or intramuscular injection. This may be accomplished by the use of
a liquid suspension of crystalline or amorphous material having
poor water solubility. The rate of absorption of the drug then
depends upon its rate of dissolution, which, in turn, may depend
upon crystal size and crystalline form. Alternatively, delayed
absorption of a parenterally administered drug form is accomplished
by dissolving or suspending the drug in an oil vehicle.
[0063] Injectable depot forms are made by forming microencapsulated
matrices of the subject compounds in biodegradable polymers such as
polylactide-polyglycolide. Depending on the ratio of drug to
polymer, and the nature of the particular polymer employed, the
rate of drug release can be controlled. Examples of other
biodegradable polymers include poly(orthoesters) and
poly(anhydrides). Depot injectable formulations are also prepared
by entrapping the drug in liposomes or microemulsions that are
compatible with body tissue.
[0064] For use in the methods of this invention, active compounds
can be given per se or as a pharmaceutical composition containing,
for example, 0.1 to 99.5% (more preferably, 0.5 to 90%) of active
ingredient in combination with a pharmaceutically acceptable
carrier.
[0065] Methods of introduction may also be provided by rechargeable
or biodegradable devices. Various slow release polymeric devices
have been developed and tested in vivo in recent years for the
controlled delivery of drugs, including proteinacious
biopharmaceuticals. A variety of biocompatible polymers (including
hydrogels), including both biodegradable and non-degradable
polymers, can be used to form an implant for the sustained release
of a compound at a particular target site.
[0066] Actual dosage levels of the active ingredients in the
pharmaceutical compositions may be varied so as to obtain an amount
of the active ingredient that is effective to achieve the desired
therapeutic response for a particular patient, composition, and
mode of administration, without being toxic to the patient.
[0067] The selected dosage level will depend upon a variety of
factors including the activity of the particular compound or
combination of compounds employed, or the ester, salt or amide
thereof, the route of administration, the time of administration,
the rate of excretion of the particular compound(s) being employed,
the duration of the treatment, other drugs, compounds and/or
materials used in combination with the particular compound(s)
employed, the age, sex, weight, condition, general health and prior
medical history of the subject being treated, and like factors well
known in the medical arts.
[0068] A physician or veterinarian having ordinary skill in the art
can readily determine and prescribe the therapeutically effective
amount of the pharmaceutical composition required. For example, the
physician or veterinarian could start doses of the pharmaceutical
composition or compound at levels lower than that required in order
to achieve the desired therapeutic effect and gradually increase
the dosage until the desired effect is achieved. By
"therapeutically effective amount" is meant the concentration of a
compound that is sufficient to elicit the desired therapeutic
effect. It is generally understood that the effective amount of the
compound will vary according to the weight, sex, age, and medical
history of the subject. Other factors which influence the effective
amount may include, but are not limited to, the severity of the
subject's condition, the disorder being treated, the stability of
the compound, and, if desired, another type of therapeutic agent
being administered with the compound of the invention. A larger
total dose can be delivered by multiple administrations of the
agent. Methods to determine efficacy and dosage are known to those
skilled in the art (Isselbacher et al. (1996) Harrison's Principles
of Internal Medicine 13 ed., 1814-1882, herein incorporated by
reference).
[0069] In general, a suitable daily dose of an active compound used
in the compositions and methods of the invention will be that
amount of the compound that is the lowest dose effective to produce
a therapeutic effect. Such an effective dose will generally depend
upon the factors described above.
[0070] If desired, the effective daily dose of the active compound
may be administered as one, two, three, four, five, six or more
sub-doses administered separately at appropriate intervals
throughout the day, optionally, in unit dosage forms. In certain
embodiments of the present invention, the active compound may be
administered two or three times daily. In preferred embodiments,
the active compound will be administered once daily.
[0071] The subject receiving this treatment is any animal in need,
including primates, in particular humans, and other mammals such as
equines, cattle, swine and sheep; and poultry and pets in
general.
[0072] In certain embodiments, compounds of the invention may be
used alone or conjointly administered with another type of
therapeutic agent. As used herein, the phrase "conjoint
administration" refers to any form of administration of two or more
different therapeutic compounds such that the second compound is
administered while the previously administered therapeutic compound
is still effective in the body (e.g., the two compounds are
simultaneously effective in the subject, which may include
synergistic effects of the two compounds). For example, the
different therapeutic compounds can be administered either in the
same formulation or in a separate formulation, either concomitantly
or sequentially. In certain embodiments, the different therapeutic
compounds can be administered within one hour, 12 hours, 24 hours,
36 hours, 48 hours, 72 hours, or a week of one another. Thus, a
subject who receives such treatment can benefit from a combined
effect of different therapeutic compounds.
[0073] This invention includes the use of pharmaceutically
acceptable salts of compounds of the invention in the compositions
and methods of the present invention. The term "pharmaceutically
acceptable salt" as used herein includes salts derived from
inorganic or organic acids including, for example, hydrochloric,
hydrobromic, sulfuric, nitric, perchloric, phosphoric, formic,
acetic, lactic, maleic, fumaric, succinic, tartaric, glycolic,
salicylic, citric, methanesulfonic, benzenesulfonic, benzoic,
malonic, trifluoroacetic, trichloroacetic, naphthalene-2-sulfonic,
and other acids.
[0074] In further embodiments, contemplated salts of the invention
include, but are not limited to, alkyl, dialkyl, trialkyl or
tetra-alkyl ammonium salts. In certain embodiments, contemplated
salts of the invention include, but are not limited to, L-arginine,
benenthamine, benzathine, betaine, calcium hydroxide, choline,
deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol,
ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine,
1H-imidazole, lithium, L-lysine, magnesium,
4-(2-hydroxyethyl)morpholine, piperazine, potassium,
1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine,
tromethamine, and zinc salts. In certain embodiments, contemplated
salts of the invention include, but are not limited to, Na, Ca, K,
Mg, Zn or other metal salts. Pharmaceutically acceptable salt forms
can include forms wherein the ratio of molecules comprising the
salt is not 1:1. For example, the salt may comprise more than one
ascr#7 molecule per molecule of base, such as two ascr#7 molecules
per molecule of compound of base. As another example, the salt may
comprise less than one ascr#7 molecule per molecule of base, such
as two molecules of ascr#7 per molecule of lysine, or per atom of
magnesium or zinc.
[0075] The pharmaceutically acceptable acid addition salts can also
exist as various solvates, such as with water, methanol, ethanol,
dimethylformamide, and the like. Mixtures of such solvates can also
be prepared. The source of such solvate can be from the solvent of
crystallization, inherent in the solvent of preparation or
crystallization, or adventitious to such solvent.
[0076] Wetting agents, emulsifiers and lubricants, such as sodium
lauryl sulfate and magnesium stearate, as well as coloring agents,
release agents, coating agents, sweetening, flavoring and perfuming
agents, preservatives and antioxidants can also be present in the
compositions.
[0077] Examples of pharmaceutically acceptable antioxidants
include: (1) water-soluble antioxidants, such as ascorbic acid,
cysteine hydrochloride, sodium bisulfate, sodium metabisulfite,
sodium sulfite and the like; (2) oil-soluble antioxidants, such as
ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated
hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol,
and the like; and (3) metal-chelating agents, such as citric acid,
ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid,
phosphoric acid, and the like.
III. Indications
[0078] In certain aspects, the invention provides methods of
preventing, alleviating, or treating an autoimmune and/or
inflammatory disease, e.g., an IL-6- and/or IL-1.beta.-mediated
disease, in a subject, comprising administering to the subject a
composition comprising ascr#7 or a pharmaceutically acceptable salt
and/or prodrug thereof.
[0079] In certain aspects, the invention provides use of ascr#7 in
the manufacture of a medicament for the treatment of an autoimmune
and/or inflammatory disease, e.g., an IL-6- and/or
IL-1.beta.-mediated disease.
[0080] Elevated levels of IL-6 and/or IL-1.beta. have been found in
patients with various diseases, and studies have shown that IL-6
and/or IL-1.beta. contribute to these diseases' pathogeneses. These
IL-6- and/or IL-1.beta.-mediated diseases may include diseases that
affect musculoskeletal or connective tissues. For example, IL-1RA,
IL-6, and IL-8 have been found to be increased in patients with
fibromyalgia (Rodriguez-Pinto et al. Immunology Letters 2014;
161:200-203). Elevated IL-6 and other inflammation markers have
been found in patients with chronic nonbacterial osteomyelitis and
chronic recurrent multifocal osteomyelitis in the absence of
high-titer auto-antibodies (Hedrich et al. Pediatric Rheumatology
2013; 11:47). Analysis of serum from patients with juvenile
dermatomyositis also showed significantly higher levels of MCP-1
and IL-6, than samples taken from healthy controls (Nistala et al.
Arthritis Research & Therapy 2013; 15:R131). IL-6 was
significantly elevated in patients with ankylosing spondylitis
compared to the controls, and serum leptin and IL-6 levels
correlated significantly with Bath Ankylosing Spondylitis Disease
Activity Index (Elolemy et al. The Egyptian Rheumatologist 2013;
35:65-70). In addition, IL-6 is likely responsible in part for Th17
activity in cutaneous lupus erythematosus (Achtman and Werth,
Arthritis Research & Therapy 2015; 17:182). IL-6 has a
significant role in the pathophysiology of active systemic lupus
erythematosus (SLE), and there is a good correlation between IL-6
and SLE disease activity (Umare et al. J Postgrad Med. 2017;
63(2):92-95). Expression of IL-1RI, and IL-1RII and their ligands,
i.e., IL-1.alpha. and IL-1.beta. was significantly higher in
patients with polymyositis and dermatomyositis than in controls,
which supports the hypothesis of a crucial role of IL-1 in the
pathogenesis of polymyositis and dermatomyositis (Grundtman et al.
ARTHRITIS & RHEUMATISM 2007; 56(2):674-687).
[0081] The IL-6- and/or IL-1.beta.-mediated diseases may include
diseases that affect renal system. For example, elevations of IL-1
and IL-6 were identified within 3 days of onset of anti-GBM
glomerulonephritis (Kim et al. Kidney International 2004;
66:1826-1837). Another study shows that IL-6 plays an important
role in the development glomerulonephritis with multinucleated
giant cells, and regression of clinical symptoms was well
correlated with a reduction in serum IL-6 levels (Ito et al. Am J
Kidney Dis. 1995; 26(1):72-9). IL-6 and histamine were
significantly greater in patients with interstitial cystitis than
in the control (Lamale et al. Adult Urology 2006; 68(4)42-46).
Higher production of IL-6, IL-8, monocyte chemotactic protein 1,
TGF-.beta. and fibronectin was found in patients with IgA
nephropathy compared to that in healthy volunteers (Liang et al.
Cell Physiol Biochem 2015; 36:1793-1808).
[0082] The IL-6- and/or IL-1.beta.-mediated diseases may be
dermatological disorders. For example, serum levels of both IL-6
and GM-CSF of the patients with both focal and generalized type of
vitiligo were significantly higher than those of controls, while
the serum level of IL-1.beta. of patients with generalized type of
vitiligo were significantly higher than those of controls and with
segmental type of vitiligo. In addition, IL-6 level of the
generalized type in progressive stage were significantly higher
than those in stable states. These findings suggest that IL-6 and
GM-CSF may be involved in the autoimmune mechanism of non-segmental
vitiligo (Tu et al. Journal of Dermatological Science 2003;
31:73-78).
[0083] The IL-6- and/or IL-1.beta.-mediated diseases may include
diseases that affect reproductive system. For example, IL-6 was
significantly higher in the serum of subjects with endometriosis
than in the control group. Therefore, serum IL-6 provides a
promising marker for nonsurgical prediction of endometriosis
(Othman et al. European Journal of Obstetrics & Gynecology and
Reproductive Biology 2008; 137:240-246).
[0084] The IL-6- and/or IL-1.beta.-mediated diseases may include
diseases that affect cardiovascular system. For example, in both
giant cell myocarditis and coxsackie virus induced viral
myocarditis, levels of IL-1.beta. and IL-6 in the pericardial
effusion were elevated compared with the serum cytokines at the
peak of inflammation (Nakayama et al. Jpn Circ J. 2000;
64(12):977-81). Increased levels of the Th17 enhancing cytokines,
TGF.beta., IL-6, and IL-23 can be found in the sera of patients
with granulomatosis with polyangiitis (Wilde et al. Arthritis
Research & Therapy 2012; 14: R227). The concentration of serum
IL-6 was also significantly increased in patients with
Henoch-Schonlein Purpura compared to controls (Su et al.
International Immunopharmacology 2016; 39:246-250). Patients with
Wegener's granulomatosis had elevated levels of IL-6 as well
(Kobold et al. Ann Rheum Dis 1999; 58:237-245). In addition, serum
IL-6 was elevated in patients with Giant Cell Arteritis and
Polymyalgia Rheumatica, and IL-6 had the strongest association with
disease activity compared to other cytokines measured in the serum
(Van der Geest et al. Rheumatology 2015; 54:1397-1402). IL-6 showed
significant correlations with the C-reactive protein (CRP) and
erythrocyte sedimentation rates in 17 children with active
rheumatic fever, and significant correlation was found between IL-6
levels and the white blood cell (WBC) counts (Yegin et al. Eur J
Pediatr. 1997; 156(1):25-9). IL-6 was elevated in patients with
Kawasaki Disease, and IL-6 is a critical cytokine in the Kawasaki
Disease pathogenesis of autoimmune vasculitis (Tan et al. Indian
Pediatrics 2013; 50:501-503).
[0085] The IL-6- and/or IL-1.beta.-mediated diseases may include
diseases that affect respiratory system. For example, IL-6 levels
were specifically elevated in asthmatic subjects compared with
healthy controls, and the relationship between asthma and lung
function could be mediated by IL-6 (Neveu et al. Respiratory
Research 2010; 11:28).
[0086] The IL-6- and/or IL-1.beta.-mediated diseases may include
diseases that affect hematological system. For example, circulating
IL-6 may persist elevated in patients with giant cell arteritis and
remain higher in patients who have experienced relapse
(Garcia-Martinez et al. Arthritis Care & Research 2010;
62(6):835-841). Serum Th2 cytokine levels such as IL-6 and IL-10
were high and Th1 cytokines such as IL2 and IFN-.gamma. were low at
onset in a 1-year-old girl with Evans syndrome coexisting with
histologically confirmed Langerhans cell histiocytosis (Tsuji et
al. Int J Hematol 2008; 87:75-77). IVIG infusion significantly
increased serum concentration of IL-6 in patients with primary
hypogammaglobulinemia (Ibanez et al. Biodrugs 2005; 19(1):59-65).
Patients with Primary Anti-Phospholipid Syndrome had higher levels
of IL-6 compared to controls, and IL-6 is known to induce the
maturation of B-cells into plasma cells, which then secrete
antibodies and contribute to the disease progress (Soltesz et al.
Rheumatology 2008; 47: 1628-1634). IL-6 is elevated in patients
with Chronic Cold Agglutinin Disease, and IL-6 plays a role in
fever induction, which is thought to lead to blood hemolysis
(Ulvestad et al. Scand. J. Immunol. 2001; 54:239-242,). In
addition, cytokine production by salivary gland epithelial cells
from Sjogren's syndrome (SS) patients showed increased IL-6 and
decreased TGF.beta. secretion. This imbalance may be critical in
the regulation of Treg/Th17 cells and may foster a pathogenic
milieu that may be causative and predictive in SS (Kawanami et al.
PLOS ONE 2012; 7(10):101-106).
[0087] The IL-6- and/or IL-1.beta.-mediated diseases may include
diseases that affect gastrointestinal system. For example, the
levels of pro-inflammatory cytokines, IL-2, IL-4, IL-6, IL-12p70,
and IFN.gamma. were increased 6-9-fold in the blood serum of
patients with IBDs compared to that from controls (Toptygina et al.
Biokhimiya, 2014; 79(12):1673-1679). IL-6 level was significantly
elevated (281.5 pg/ml in the new/active EoE patient group compared
to 16.2 pg/ml in the control group) in biopsy culture supernatant
from esophageal samples from patients with eosinophilic esophagitis
(Sayej et al. Clinical & Translational Immunology 2016;
5:25-36). The average serum IL-6 level in the EoE patients is also
at least two-fold higher than the level in the healthy controls
(Blanchard et al. J Allergy Clin Immunol. 2011; 127(1):208-17). In
addition, the IL-1.beta. gene was also significantly elevated
(about 7-fold using microarray analysis and about 5.5-fold using
real-time PCR) in biopsies from esophageal samples from patients
with eosinophilic esophagitis (Blanchard et al. J Allergy Clin
Immunol. 2011; 127(1):208-17). There was also a significant
increase in median IL-6 staining in both the lamina propria and the
epithelium of untreated celiac disease patients, when compared with
treated celiac disease patients (Przemioslo et al. Gut 1994;
35:1398-1403). In addition, Type I diabetic children living in
moderate altitude or at sea level showed elevated levels of IL-6,
along with other pro-inflammatory cytokines (Allam et al. High
Altitude Medicine & Biology 2014; 15(3):59-66). Patients with
type 1 diabetes had higher levels of mannose binding lectin (MBL)
compared to siblings of these patients, and increased MBL levels
positively correlated with IL-1.beta., IL-12, and IL-18 (Sildorf et
al. Molecular Immunology 2014; 62:71-76). The hepatic expression of
IL-17, IL-23, IL-6, and IL-1.beta., etc. in patients with
autoimmune hepatitis were also significantly increased and were
associated with increased inflammation and fibrosis (Zhao et al.
PLOS ONE 2011; 6(4):124-131). Muramyl dipeptides, the minimal
NOD2/CARD15 (mutations in which increases the susceptibility for
Crohn's disease) activating component of peptidoglycan induced
IL-1.beta. and IL-8 transcripts (Li et al. Human Molecular
Genetics, 2004; 13(16):1715-1725).
[0088] The IL-6- and/or IL-1.beta.-mediated diseases may include
diseases that physically affect the nervous system. For example,
levels of IL-6 among the transverse myelitis patient population
directly correlated with markers of tissue injury and the severity
of paralysis (Kaplin et al. J Clin Invest. 2005; 115(10):2731-41).
IL-2 and IL-6 were also increased in patients with Guillain-Barre
syndrome (Creange et al. J Neurol Neurosurg Psychiatry 1998;
64:162-165). In addition an IL-6 receptor antibody has been found
to treat AA amyloidosis (Ando, Rinsho Shinkeigaku (Cln Neurol)
2015; 55:797-803).
[0089] In addition, steroid-resistant patients with autoimmune
inner ear disease have increased expression of IL-1.beta. (Rauch S
D, J Clin Invest. 2014; 124(9):3685-3687).
[0090] The IL-6- and/or IL-1.beta.-mediated diseases may include
diseases that affect endocrine system. For example, IL-6 was
elevated in patient group with Graves' disease (Niyazoglu et al.
Gene 547 (2014) 226-232). Levels of IL-6 was also significantly
increased in Hashimoto thyroiditis patients compared to the control
group, and higher IL-6 levels are associated with increased risk
factor in Hashimoto thyroiditis (Koc et al. Immunobiology 2014;
219:531-536). IL-6 was markedly increased in Graves' disease
patients compared to normal controls (Al-Humaidi M A, Saudi Medical
Journal 2000; 21(7):639-644).
[0091] The IL-6- and/or IL-1.beta.-mediated diseases may include
diseases that affect multiple tissues or organs systematically. For
example, IL-6 is associated with diabetes, Crohn's disease, and
rheumatoid arthritis (Maggio et al. J Gerontol A Biol Sci Med Sci.
2006; 61(6): 575-584).
[0092] As IL-6 and/or IL-1.beta. plays an important role in many
autoimmune and inflammatory diseases, targeting IL-6 and/or
IL-1.beta. is being used as a therapy for these diseases. For
example, IL6 receptor antibody has been found to be effective in
treating Amyloid A (AA) amyloidosis (Ando Y., Rinsho Shinkeigaku.
2015; 55(11):797-803). With the treatment of humanized anti-IL-6
receptor antibody, the serum hemoglobin level of patients with
autoimmune hemolytic anemia gradually increased and normalized
within 4 months (Kunitomi et al. Int J Hematol. 2004;
80:246-249.).
[0093] Tocilizumab, a humanized anti-IL-6 receptor antibody, has
been approved as a biological drug for the treatment of rheumatoid
arthritis, Castleman's disease and systemic juvenile idiopathic
arthritis, and is expected to be applicable to various other
autoimmune and chronic inflammatory diseases (Tanaka et al. Int J
Biol Sci 2012; 8(9):1227-1236.). Other inflammatory diseases for
which Tocilizumab is therapeutically effective include SLE, adult
onset Still disease, Takayasu arthritis, polyarteritis nodosa,
systemic sclerosis, reactive arthritis, dermatomyositis, and
polymyositis (Murakami et al. Current Opinion in Rheumatology 2011;
23:273-277). In addition, significant elevation of IL-6 has been
found in ocular fluids derived from refractory/chronic uveitis
patients, and Tocilizumab has been shown to be effective in
treating uveitis (Mesquida et al. British Society for Immunology,
Clinical and Experimental Immunology 2014; 176:301-309). IL-6 serum
levels were also significantly increased in patients with
ANCA-associated vasculitis, and treatment with Tocilizumab was able
to induce a complete and sustained disease remission in a patient
with severe multisystemic microscopic polyangiitis (Berti et al.
Seminars in Arthritis and Rheumatism 2015; 45:48-54). Inflammatory
response and quality of life measures were successfully improved by
Tocilizumab of a 69-year-old man with systemic manifestations of
Cogan's syndrome relapsed after prednisone induced remission for 1
year (Shibuya et al. Mod Rheumatol 2013; 23:577-581). After one
year of treatment with Tocilizumab, subjects with neuromyelitis
optica experienced decreased relapse rates, neuropathic pain, and
general fatigue (Araki et al. Neurology 2014; 82:1302-1306). The
use of Tocilizumab allowed for prednisone withdrawal at 3 months,
and remission of both articular and cutaneous manifestations of
eosinophilic fasciitis at 6 months (Espinoza et al. Joint Bone
Spine 2015; 82:460-461). Atopic dermatitis is a multifactorial
disorder associated with Th2 polarized CD4+ T cells that can
overproduce IL-6. Blocking IL-6 receptor by Tocilizumab in 3
patients with severe atopic dermatitis decreased clinical activity
of atopic dermatitis (Navarini et al. J Allergy Clin Immunol
2011:1128-1130).
[0094] Similarly, targeting IL-1.beta. is also being used as a
therapeutic intervention for immune disorders (Dinarello et al. Nat
Rev Drug Discov. 2012; 11(8):633-652). For example, IL-1 promotes
the production of IL-6, a key pathogenic factor for Castleman's
disease. In the case of a 61-year-old woman with Castleman's
disease refractory to treatments including cladribine, rituximab,
steroids, etanercept, and anti-IL-6 monoclonal antibody, her
symptoms markedly improved with recombinant IL-1 receptor
antagonist (El-Osta et al. Mol Cancer Ther. 2010;
9(6):1485-1488).
[0095] The IL-6- and/or IL-1.beta.-mediated diseases that can be
treated with a composition comprising ascr#7 or a pharmaceutically
acceptable salt and/or prodrug thereof include but are not limited
to the diseases discussed herein.
[0096] In some embodiments, the IL-6- and/or IL-1.beta.-mediated
disease is Agammaglobulinemia, Amyloidosis, Ankylosing spondylitis,
Anti-GBM/Anti-TBM nephritis, Antiphospholipid syndrome, Autoimmune
hepatitis, Autoimmune inner ear disease, Atopic dermatitis, Asthma,
Castleman disease, Celiac disease, Chagas disease, Chronic
recurrent multifocal osteomyelitis, Cogan's syndrome, Cold
agglutinin disease, CREST syndrome, Crohn's disease,
Dermatomyositis, Devic's disease (neuromyelitis optica), Discoid
lupus, Endometriosis, Eosinophilic esophagitis, Eosinophilic
fasciitis, Evan's syndrome, Fibromyalgia, Giant cell arteritis,
Giant cell myocarditis, Glomerulonephritis, Goodpasture's syndrome,
Granulomatosis with polyangiitis, Graves' disease, Guillain-Barre
syndrome, Hashimoto's thyroiditis, Hemolytic anemia,
Henoch-Schonlein purpura, Hypogammaglobulinemia, Hypoproliferative
anemia, IgA Nephropathy, Inclusion body myositis, Interstitial
cystitis, Inflammatory Bowel Disease, Juvenile arthritis,
Juvenile/Type 1 Diabetes, Juvenile myositis, Kawasaki syndrome,
Lichen planus, Lichen sclerosus, Lupus (SLE), Meniere's disease,
Multiple sclerosis, Myasthenia gravis, Microscopic polyangiitis,
Optic neuritis, Pemphigus, Polyarteritis nodosa, Polymyalgia
rheumatica, Polymyositis, Primary biliary cirrhosis, Primary
sclerosing cholangitis, Psoriasis, Psoriatic arthritis, Rheumatic
fever, Rheumatoid arthritis, Sarcoidosis, Sjogren's syndrome,
Temporal arteritis/Giant cell arteritis, Transverse myelitis,
Ulcerative colitis, Uveitis, Vasculitis, Vitiligo, Viral
myocarditis, or Wegener's granulomatosis (Granulomatosis with
Polyangiitis (GPA)).
[0097] In certain embodiments, the disease is asthma, inflammatory
bowel disease, or type 1 diabetes. In certain preferred
embodiments, the disease is Eosinophilic esophagitis (EoE).
[0098] The invention also provides methods of reducing IL-6 and/or
IL-1.beta. production from a cell, comprising contacting the cell
with a composition comprising ascr#7 or a pharmaceutically
acceptable salt or prodrug thereof. Such methods may be performed
in vivo or in vitro.
IV. Subjects
[0099] The subject may be a mammal. The subject may be a rodent,
lagomorph, feline, canine, porcine, ovine, bovine, equine, or
primate. In some embodiments, the subject is a human. The subject
may be a female or male. The subject may be an infant, child, or
adult. In some embodiments, the subject has an elevated level of
IL-6 and/or IL-1.beta..
V. ROUTES OF Administration
[0100] A pharmaceutical composition (preparation) can be
administered to a subject by any of a number of routes of
administration including, for example, orally (for example,
drenches as in aqueous or non-aqueous solutions or suspensions,
tablets, capsules (including sprinkle capsules and gelatin
capsules), boluses, powders, granules, pastes for application to
the tongue); absorption through the oral mucosa (e.g.,
sublingually); anally, rectally or vaginally (for example, as a
pessary, cream or foam); parenterally (including intramuscularly,
intravenously, subcutaneously or intrathecally as, for example, a
sterile solution or suspension); nasally; intraperitoneally;
subcutaneously; transdermally (for example as a patch applied to
the skin); and topically (for example, as a cream, ointment or
spray applied to the skin, or as an eye drop). The compound may
also be formulated for inhalation. In certain embodiments, a
compound may be simply dissolved or suspended in sterile water.
Details of appropriate routes of administration and compositions
suitable for same can be found in, for example, U.S. Pat. Nos.
6,110,973, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and
4,172,896, as well as in patents cited therein.
[0101] One of skill in the art would appreciate that a method of
administering a therapeutically effective substance formulation or
composition of the disclosure would depend on factors such as the
age, weight, and physical condition of the subject being treated,
and the disease or condition being treated. The skilled worker
would, thus, be able to select a method of administration optimal
for a subject on a case-by-case basis.
[0102] The invention now being generally described, it will be more
readily understood by reference to the following examples, which
are included merely for purposes of illustration of certain aspects
and embodiments of the present invention, and are not intended to
limit the invention. As such, it will be readily apparent that any
of the disclosed beneficial substances and therapies can be
substituted within the scope of the present disclosure.
EXAMPLES
[0103] Many types of autoimmune and inflammatory diseases, have
shared components of their disease mechanism. One of the main
shared components involves signaling molecules, known as cytokines,
which guide and direct many cellular processes. IL-6 and/or
IL-1.beta. are found to be significantly elevated in many
autoimmune and/or inflammatory disorders.
[0104] The Examples variously demonstrate that ascr#7 significantly
reduced levels of both IL-6 and IL-1.beta. in an in-vitro model of
inflammation. They further demonstrate that ascr#7 significantly
reduces disease progression in a broad range of autoimmune and
inflammatory diseases, specifically a mouse model of asthma, a
mouse model of type 1 diabetes, and a mouse model of inflammatory
bowel disease.
Example 1
[0105] Ascr#7 significantly reduces both IL-6 and IL-1.beta.
secretion in an in vitro model of inflammation. Specifically,
ascr#7 reduces IL-6 and IL-1.beta. in a stimulated culture of bone
marrow-derived mouse dendritic cells.
[0106] a. Generation of Bone Marrow Dendritic Cells
[0107] Bone marrow was extracted from femurs of C57BL/6 mice (6-12
weeks of age). These mice were euthanized by CO.sub.2 inhalation
and their femurs were removed. After removing the remaining flesh
from the bone, they were cut at the metaphysis. Bone marrow was
flushed out using a syringe filled with RPMI. The extracted marrow
was further broken down by resuspension with syringe. The mixture
was centrifuged and the pellet was re-suspended in RPMI. Cells from
the bone marrow were seeded in 6 wells such that each well had a
density of 10.sup.6 cells/ml. 20 ng of GM-CSF was added to each
well. At day 3, and every 3 days thereafter, the media was
replaced. Cells were used for experiments between day 7-9.
[0108] b. Stimulation of Dendritic Cells: Testing the Effect of
Ascr#7
[0109] Bone marrow dendritic cells were collected between day 7-9
of culture. Cells were washed twice and counted. Ascr#7 in water or
RPMI were placed in corresponding wells, and are allowed to dry.
10.sup.6 cells were then plated in each well in a 24 well culture
plate. Cells were left to incubate with ascr#7 for 5 minutes.
Following the brief incubation, different TNF.alpha. was added to
the wells. These bone marrow dendritic cells were then cultured at
37.degree. C. for 18-72 hours. Cells were collected via the pellet
generated by centrifuge. The supernatant was also preserved.
EBioscience ELISA kits was used to determine the concentration of
different cytokines present in each well. Results are shown in FIG.
1 and FIG. 2.
[0110] The left side of FIG. 1 shows that the dendritic cells, when
not stimulated with TNF-.alpha., have very low levels of IL-6, and
that this remains true with administration of different
concentrations of ascr#7. The right side of FIG. 1 shows that
dendritic cells, when stimulated with TNF-.alpha., has very high
levels of secreted IL-6 and that administration of different
concentrations of ascr#7 reduces these levels significantly. The
left side of FIG. 2 shows that the dendritic cells, when not
stimulated with TNF-.alpha., have very low levels of IL-1.beta.,
and that this remains true with administration of different
concentrations of ascr#7. The right side of FIG. 2 shows that
dendritic cells, when stimulated with TNF-.alpha., has very high
levels of secreted IL-.beta. and that administration of different
concentrations of ascr#7 reduces these levels significantly.
Example 2
[0111] Ascr#7 reduces the trademark disease pathologies in a mouse
model of asthma. Specifically, ascr#7 reduces the development of
eosinophilia, goblet cell metaplasia, and airway hyper-reactivity
in the ovalbumin model of murine asthma.
[0112] Adult C57BL/6 mice were sensitized by administering 2 ug OVA
(Sigma-Aldrich) and 1 mg alum (Thermo) with 50 ng ascr#7 or saline,
via intraperitoneal injection, on Day 0 and 14. On day 24, the mice
were challenged with intranasal administration of saline containing
100 .mu.g OVA, under isoflurane anesthesia. Histologic sections
were stained with H&E or PAS to score for eosinophilia, goblet
cell metaplasia, and airway hyper-reactivity. Results are shown in
FIG. 3B and FIG. 3C. Mice were also placed in a computer controlled
small animal ventilator (SCIREQ) to score for airway
hyper-reactivity following methacholine challenge. FIG. 3A shows a
diagram of a mouse in a respiratory chamber, which has been
calibrated to calculate the resistance of air flow as the animal
breathes. Animals with asthma have higher resistance of flow, which
would be represented as higher "RL" values in FIG. 3B. FIG. 3B
shows that asthmatic mice, here labeled "Control", have higher
resistance of air flow, whereas a normal mouse, here labeled "No
Immunization" has low resistance of air flow. Asthmatic mice that
have been administered ascr#7 demonstrate low resistance of air
flow, with comparable levels to normal mice. Lungs were collected,
postmortem, and then sectioned and stained to show the presence of
eosinophils and mucus. These findings are shown in FIG. 3C. In FIG.
3C, the normal mouse lung, here labeled "No Immunization" shows
thin boundaries between the central bronchioles and the surrounding
alveoli, wherein the thin membranes allow for oxygen to easily
diffuse. In FIG. 3C, the asthmatic mouse lung, here labeled
"Control" has a large number of mucus and eosinophils that surround
the bronchioles and occupy the alveolar space; wherein the thick
barriers would prohibit oxygen from easily diffusing. In FIG. 3C,
the lung of asthmatic mice that were administered ascr#7 has thin
boundaries between the bronchioles and the alveolar space,
comparable to the normal mice. This demonstrates that ascr#7
reduces the trademark pathologies found in asthmatic mice,
specifically resistance to air flow and its associated
histopathologies.
Example 3
[0113] Ascr#7 reduces the signature disease pathologies in a mouse
model of Type 1 Diabetes. Specifically, ascr#7 delays conversion to
hyperglycemia in a multi-genetic animal model of Type 1
Diabetes.
[0114] The NOD/ShiLtJ (Jackson Laboratories) mouse model of Type 1
Diabetes was used These mice typically develop hyperglycemia around
17-18 weeks of life. Either saline or 100 ng ascr#7 was
administered three times a week via intraperitoneal injection.
Blood glucose levels were determined in the same mouse over time,
via the collected of tail vein blood (as shown in FIG. 4A) at
multiple time points. FIG. 4B shows the individual blood glucose
values of 16 NOD/ShiLtJ mice over 18 weeks. 8 mice were injected
with saline and 8 mice were injected with ascr#7, as described
above. FIG. 4B demonstrates that most of the untreated mice,
injected with saline (blank circles), indeed had high blood glucose
values by week 18. FIG. 4B demonstrates that most of the mice
treated with ascr#7 (solid circles) had significantly reduced blood
glucose levels throughout 18 weeks. FIG. 4C shows the combined
average and statistical error bars of the mice shown in FIG. 4B.
FIG. 4C shows that at week 18, the mice treated with ascr#7 had
significantly lower blood glucose levels compared to that of the
untreated group.
Example 4
[0115] Ascr#7 reduces the signature disease pathologies in a mouse
model of IBD. Specifically, ascr#7 markedly reduces the development
of colon shortening, abnormal lumen dilation, and diarrhea in an
immune dysregulation model of inflammatory bowel disease.
[0116] A T cell transfer model of colitis where 10.sup.6CD4+ T
cells were transferred to adult Rag -/- mice was used, and either
saline or 300 ng ascr#7 were administered via intraperitoneal
injection every 3 days between day 14 and day 28. On day 28, the
mice were euthanized and colon analysis was performed. n=3 mice per
group. Results are shown in FIG. 5. The inset shows a diagram of
the colon. FIG. 5 shows the normal tissue architecture of a
dissected caecum and colon. The normal mouse colon is long, skinny,
and has discrete fecal pellets. The diseased mouse colon, with IBD,
has a shortened, dilated colon, with diarrhea instead of discrete
fecal pellets. The colon of a mouse with IBD, which has been
administered ascr#7 has a normal appearing colon, which is long,
skinny, and contains discrete fecal pellets.
INCORPORATION BY REFERENCE AND VARIATIONS
[0117] All publications and patents mentioned herein are hereby
incorporated by reference in their entirety as if each individual
publication or patent was specifically and individually indicated
to be incorporated by reference. In case of conflict, the present
specification, including its specific definitions, will
control.
[0118] While specific aspects of the subject matter have been
discussed, the above specification is illustrative and not
restrictive. Many variations will become apparent to those skilled
in the art upon review of this specification and the claims below.
The full scope of the disclosure should be determined by reference
to the claims, along with their full scope of equivalents, and the
specification, along with such variations.
* * * * *