U.S. patent application number 13/388411 was filed with the patent office on 2012-05-31 for treatment of macrophage-related disorders.
Invention is credited to Arasteh Ari Azhir, Michael S. McGrath.
Application Number | 20120134929 13/388411 |
Document ID | / |
Family ID | 43544843 |
Filed Date | 2012-05-31 |
United States Patent
Application |
20120134929 |
Kind Code |
A1 |
McGrath; Michael S. ; et
al. |
May 31, 2012 |
TREATMENT OF MACROPHAGE-RELATED DISORDERS
Abstract
The present invention provides a method of treating a macrophage
related disease comprising administering to a subject in need
thereof an effective amount of an oxidative agent or an
immunosuppressive agent. The present invention also provides a
method of modulating macrophage accumulation or activation
comprising administering to a subject in need thereof an effective
amount of an oxidative agent or an immunosuppressive agent. The
oxidative agent can be chlorite or a chlorite containing
compound.
Inventors: |
McGrath; Michael S.;
(Burlingame, CA) ; Azhir; Arasteh Ari; (Los Altos,
CA) |
Family ID: |
43544843 |
Appl. No.: |
13/388411 |
Filed: |
July 23, 2010 |
PCT Filed: |
July 23, 2010 |
PCT NO: |
PCT/US10/43150 |
371 Date: |
February 1, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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61231989 |
Aug 6, 2009 |
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61232678 |
Aug 10, 2009 |
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61238609 |
Aug 31, 2009 |
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Current U.S.
Class: |
424/9.2 ;
424/665; 435/7.92; 436/501; 514/390; 514/604 |
Current CPC
Class: |
A61P 11/06 20180101;
A61P 25/00 20180101; A61K 31/10 20130101; A61K 45/06 20130101; A61K
31/137 20130101; A61K 31/7076 20130101; A61K 39/3955 20130101; C07K
16/2839 20130101; A61K 33/18 20130101; A61P 25/16 20180101; A61P
1/02 20180101; A61P 9/12 20180101; A61K 33/40 20130101; A61K 31/327
20130101; A61K 31/185 20130101; Y02A 50/409 20180101; A61P 9/10
20180101; G01N 2333/70596 20130101; A61K 33/16 20130101; A61P 25/28
20180101; A61P 35/00 20180101; Y02A 50/30 20180101; A61P 3/04
20180101; A61K 33/38 20130101; A61P 3/10 20180101; A61K 33/00
20130101; A61K 33/04 20130101; A61K 33/32 20130101; A61P 11/00
20180101; A61P 37/06 20180101; A61K 33/24 20130101; A61P 7/00
20180101; A61P 43/00 20180101; A61P 31/00 20180101; A61K 33/20
20130101; A61P 29/00 20180101; A61P 33/02 20180101; A61P 21/00
20180101; A61K 31/10 20130101; A61K 2300/00 20130101; A61K 31/185
20130101; A61K 2300/00 20130101; A61K 31/327 20130101; A61K 2300/00
20130101; A61K 33/00 20130101; A61K 2300/00 20130101; A61K 33/04
20130101; A61K 2300/00 20130101; A61K 33/16 20130101; A61K 2300/00
20130101; A61K 33/18 20130101; A61K 2300/00 20130101; A61K 33/20
20130101; A61K 2300/00 20130101; A61K 33/24 20130101; A61K 2300/00
20130101; A61K 33/32 20130101; A61K 2300/00 20130101; A61K 33/38
20130101; A61K 2300/00 20130101; A61K 33/40 20130101; A61K 2300/00
20130101 |
Class at
Publication: |
424/9.2 ;
424/665; 514/390; 514/604; 436/501; 435/7.92 |
International
Class: |
A61K 33/00 20060101
A61K033/00; A61K 31/18 20060101 A61K031/18; G01N 33/53 20060101
G01N033/53; A61P 3/10 20060101 A61P003/10; A61P 25/28 20060101
A61P025/28; A61P 25/16 20060101 A61P025/16; A61K 31/4166 20060101
A61K031/4166; A61K 49/00 20060101 A61K049/00 |
Claims
1. A method of reducing the side effects of an immunomodulator
administered to a subject in need thereof, said method comprising
administering an oxidative agent to said subject in combination
with said immunomodulator.
2. The method of claim 1, wherein said immunomodulator is
administered for the treatment of a macrophage-related disorder
selected from the group consisting of cancer, multiple sclerosis,
rheumatoid arthritis, macrophage activation syndrome,
atherosclerosis, Type I diabetes, Type II diabetes, Kawasaki
disease, asthma, hemophagocytic lymphohistiocytosis, sarcoidosis,
periodontitis, Whipple's disease, pulmonary alveolar proteinosis,
macrophage-related pulmonary disease, Leishmaniasis,
hemodialysis-related inflammation, microbial infection,
transplant-related complications, metabolic syndrome, hypertension,
and inflammatory neurological diseases.
3. The method of claim 1, wherein said immunomodulator is selected
from the group consisting of immunosuppressants, macrophage
migration inhibitors, anti-inflammatory agents, and
immunomodulatory antibodies.
4. The method of claim 1, wherein said oxidative agent is selected
from i) non-halogen activated-oxygen compounds selected from
potassium nitrate (KNO.sub.3), permanganate salts, ammonium
cerium(IV) nitrate, hexavalent chromium compounds,
chromate/dichromate compounds, ammonium silver nitrate, sulfoxides,
persulfuric acid, osmium tetroxide (OsO.sub.4), nitric acid,
nitrous oxide (N.sub.2O), hydrogen peroxide, organic peroxides,
superoxides, and ozone; ii) non-oxygen activated-halogen compounds
selected from fluorine, chlorine, bromine, and iodine; iii)
activated-halogen activated-oxygen compounds selected from sodium
chlorite and tetrachlorodecaoxygen; and iv) N-halo compounds
selected from the group consisting of N-halophthalimide,
N-halosuccinimide, N-halosaccharin, N,N-dihalourethane,
N-haloacetanilide, 1,3-dihalo-5,5-dimethylhydantoin,
trihaloisocyanuric acid, and sodium dihaloisocyanurate.
5. The method of claim 1, wherein said oxidative agent is sodium
chlorite.
6. A method of potentiating the effect of an immunomodulator
administered to a subject in need thereof, said method comprising
administering an oxidative agent to said subject in combination
with said immunomodulator.
7. The method of claim 6, wherein said immunomodulator is
administered for the treatment of a macrophage-related disorder
selected from the group consisting of cancer, multiple sclerosis,
rheumatoid arthritis, macrophage activation syndrome,
atherosclerosis, Type I diabetes, Type II diabetes, Kawasaki
disease, asthma, hemophagocytic lymphohistiocytosis, sarcoidosis,
periodontitis, Whipple's disease, pulmonary alveolar proteinosis,
macrophage-related pulmonary disease, Leishmaniasis,
hemodialysis-related inflammation, microbial infection,
transplant-related complications, metabolic syndrome, hypertension,
and inflammatory neurological diseases.
8. The method of claim 6, wherein said immunomodulator is selected
from the group consisting of immunosuppressants, macrophage
migration inhibitors, anti-inflammatory agents, and
immunomodulatory antibodies.
9. The method of claim 6, wherein said oxidative agent is selected
from i) non-halogen activated-oxygen compounds selected from
potassium nitrate (KNO.sub.3), permanganate salts, ammonium
cerium(IV) nitrate, hexavalent chromium compounds,
chromate/dichromate compounds, ammonium silver nitrate, sulfoxides,
persulfuric acid, osmium tetroxide (OsO.sub.4), nitric acid,
nitrous oxide (N.sub.2O), hydrogen peroxide, organic peroxides,
superoxides, and ozone; ii) non-oxygen activated-halogen compounds
selected from fluorine, chlorine, bromine, and iodine; iii)
activated-halogen activated-oxygen compounds selected from sodium
chlorite and tetrachlorodecaoxygen; and iv) N-halo compounds
selected from the group consisting of N-halophthalimide,
N-halosuccinimide, N-halosaccharin, N,N-dihalourethane,
N-haloacetanilide, 1,3-dihalo-5,5-dimethylhydantoin,
trihaloisocyanuric acid, and sodium dihaloisocyanurate.
10. The method of claim 6, wherein said oxidative agent is sodium
chlorite.
11. A method of treating a disease associated with migration of
activated macrophages, said method comprising administering a
therapeutically-effective amount of an oxidative agent to a subject
in need thereof, wherein said oxidative agent is selected from the
group consisting of non-halogen activated-oxygen compounds,
non-oxygen activated-halogen compounds, activated-halogen
activated-oxygen compounds, and N-halo compounds.
12. The method of claim 11, wherein i) the non-halogen
activated-oxygen compounds are selected from potassium nitrate
(KNO.sub.3), permanganate salts, ammonium cerium(IV) nitrate,
hexavalent chromium compounds, chromate/dichromate compounds,
ammonium silver nitrate, sulfoxides, persulfuric acid, osmium
tetroxide (OsO.sub.4), nitric acid, nitrous oxide (N.sub.2O),
hydrogen peroxide, organic peroxides, superoxides, and ozone; ii)
the non-oxygen activated-halogen compounds are selected from
fluorine, chlorine, bromine, and iodine; iii) the activated-halogen
activated-oxygen compounds are selected from sodium chlorite and
tetrachlorodecaoxygen; and iv) the N-halo compounds are selected
from the group consisting of N-halophthalimide, N-halosuccinimide,
N-halosaccharin, N,N-dihalourethane, N-haloacetanilide,
1,3-dihalo-5,5-dimethylhydantoin, trihaloisocyanuric acid, and
sodium dihaloisocyanurate.
13. The method of claim 11, wherein said oxidative agent is sodium
chlorite.
14. A method of treating a disease associated with excess
activation of monocytes to activated macrophages, said method
comprising administering a therapeutically-effective amount of an
oxidative agent to a subject in need thereof, wherein said
oxidative agent is selected from the group consisting of
non-halogen activated-oxygen compounds, non-oxygen
activated-halogen compounds, and N-halo compounds.
15. The method of claim 14, wherein i) the non-halogen
activated-oxygen compounds are selected from potassium nitrate
(KNO.sub.3), permanganate salts, ammonium cerium(IV) nitrate,
hexavalent chromium compounds, chromate/dichromate compounds,
ammonium silver nitrate, sulfoxides, persulfuric acid, osmium
tetroxide (OsO.sub.4), nitric acid, nitrous oxide (N.sub.2O),
hydrogen peroxide, organic peroxides, superoxides, and ozone; ii)
the non-oxygen activated-halogen compounds are selected from
fluorine, chlorine, bromine, and iodine; and iii) the N-halo
compounds are selected from the group consisting of
N-halophthalimide, N-halosuccinimide, N-halosaccharin,
N,N-dihalourethane, N-haloacetanilide,
1,3-dihalo-5,5-dimethylhydantoin, trihaloisocyanuric acid, and
sodium dihaloisocyanurate.
16. The method of claim 14, wherein said oxidative agent is
selected from 1,3-dichloro-5,5-dimethylhydantoin and
chloramine-T.
17. The method of claim 14, wherein said disease is associated with
excess CD14CD16 expression.
18. The method of claim 14, wherein said disease is a
neurodegenerative disease selected from the group consisting of
amyotrophic lateral sclerosis (ALS), Parkinson's Disease (PD),
Alzheimer's Disease (AD), and complications thereof.
19. A sodium chlorite compound, wherein said compound is a
crystalline solid of greater than 95% purity.
20. The compound of claim 19, wherein said compound is a
crystalline solid of greater than 99% purity.
21. The compound of claim 19, wherein said compound has an x-ray
powder diffraction pattern with peaks expressed in degrees 20 at
about 21, 30, 31, 32, 34, and 39.
22. A pharmaceutical composition comprising one or more
pharmaceutical excipients and the compound of claim 19, wherein
said composition is a solid.
23. A pharmaceutical composition comprising: (a) the compound of
claim 19; (b) a pH adjusting agent; and (c) a pharmaceutically
acceptable excipient or carrier, wherein said composition is a
liquid that exhibits 25% less pH drift compared to an identical
composition without said pH adjusting agent.
24. The composition of claim 23, wherein said pH adjusting agent is
sodium phosphate dibasic.
25. A method of treating Type II diabetes or related complications
comprising administering to a subject in need thereof an effective
amount of a pharmaceutical composition comprising chlorite or a
chlorite-containing agent.
26. A method of treating a disease associated with migration of
monocytes or activated macrophages, said method comprising
administering a therapeutically-effective amount of an oxidative
agent to a subject in need thereof, wherein said oxidative agent is
selected from the group consisting of non-halogen activated-oxygen
compounds, non-oxygen activated-halogen compounds,
activated-halogen activated-oxygen compounds, and N-halo
compounds.
27. The method of claim 26, wherein said disease is characterized
by elevated CD16 expression levels in CD14+ cells.
28. The method of claim 27, wherein said oxidative agent is
selected from sodium chlorite, 1,3-dichloro-5,5-dimethylhydantoin
and chloramine-T.
29. The method of claim 26, wherein said disease is characterized
by migration of PBMCs in response to chemoattractant.
30. The method of claim 29, wherein said oxidative agent is sodium
chlorite.
31. A method of treating a disease associated with excess
production of sCD14 and/or sCD163 by activated macrophages, said
method comprising administering a therapeutically-effective amount
of an oxidative agent to a subject in need thereof, wherein said
oxidative agent is selected from the group consisting of
non-halogen activated-oxygen compounds, non-oxygen
activated-halogen compounds, activated-halogen activated-oxygen
compounds, and N-halo compounds.
32. The method of claim 31, wherein said oxidative agent is sodium
chlorite.
33. A method of diagnosing a macrophage related disease in a
subject comprising measuring the level of a biomarker in the
subject and correlating the measured level of biomarker to normal
and diseased levels of said biomarker, wherein said biomarker is
selected from CD16 expression in CD14+ cells, sCD14, sCD163,
expression of chemoattractants by macrophages, and combinations
thereof.
34. A method of determining efficacy of treatment with an oxidative
agent for a macrophage related disease in a subject comprising: i)
initiating treatment with an oxidative agent; ii) measuring the
level of a biomarker in the subject; and iii) correlating the
measured level of biomarker to normal and diseased levels of said
biomarker and/or levels of biomarker in said subject prior to
treatment; wherein said biomarker is selected from CD16 expression
in CD14+ cells, sCD14, sCD163, expression of chemoattractants by
macrophages, and combinations thereof.
35. The method of claim 34, wherein said oxidative treatment is
sodium chlorite.
Description
[0001] This application claims benefit under 35 U.S.C. .sctn.119(e)
to Provisional Application Nos. U.S. 61/231,989, filed Aug. 6,
2009, U.S. 61/232,678, filed Aug. 10, 2009, and U.S. 61/238,609,
filed Aug. 31, 2009, each of which is herein incorporated by
reference in its entirety.
BACKGROUND OF THE INVENTION
[0002] Macrophages are white blood cells produced by the division
of monocytes. Monocytes and macrophages are phagocytes, and play a
role in innate immunity (non-specific immune defenses) as well as
helping to initiate adaptive immunity (specific defense
mechanisms). These cells phagocytose (engulf and then digest)
cellular debris and pathogens either as stationary or as mobile
cells. When activated by pathogens or by other mechanisms,
macrophages stimulate and recruit lymphocytes and other immune
cells to respond to the insult.
[0003] Although macrophages play a vital role in host immune
defenses, activated macrophages are also involved in the
progression of a number of diseases and disorders. Activated
macrophages elicit massive leukocyte infiltration and flood the
surrounding tissue with inflammatory mediators, pro-apoptotic
factors, and matrix degrading proteases. These actions can result
in inflammation that can dismantle tissues to the point of
inflicting serious injury. Tissue destruction perpetrated by
macrophage-induced inflammation has been associated with the
development of tumors, autoimmune disorders, and other
conditions.
[0004] Oxidative agents such as chlorite can return macrophages to
their inactivated state. Immunosuppressant agents can mitigate
macrophage activation. The present invention provides methods for
the treatment of macrophage-related diseases and related conditions
with oxidative agents or immunosuppressant agents.
SUMMARY OF THE INVENTION
[0005] In one aspect, the present invention provides a method of
treating a macrophage related disease comprising administering to a
subject in need thereof an effective amount of a pharmaceutical
composition comprising an oxidative agent.
[0006] In another aspect, the present invention provides a method
of treating a macrophage related disease comprising administering
to a subject in need thereof an effective amount of a
pharmaceutical composition comprising: chlorite; a pH adjusting
agent; and a pharmaceutically acceptable excipient. In some
embodiments, the pH adjusting agent comprises monosodium phosphate
and/or disodium phosphate. In some embodiments, the pH of the
composition is between about 7.1 and about 7.7. In some
embodiments, the weight ratio of chlorite:chlorate is greater than
100:1.5.
[0007] In still another aspect, the present invention provides a
method of treating a macrophage related disease comprising
administering to a subject in need thereof an effective amount of a
pharmaceutical composition comprising an immunomodulatory
agent.
[0008] In a further aspect, the present invention provides a method
of modulating macrophage accumulation or activation comprising
administering to a subject in need thereof an effective amount of a
pharmaceutical composition comprising an oxidative agent. A method
is also provided to modulate macrophage accumulation or activation
comprising administering to a subject in need thereof an effective
amount of a pharmaceutical composition comprising an
immunomodulatory agent. In some embodiments, the macrophage
activation is enhanced. In some embodiments, the macrophage
activation is reduced or inhibited.
[0009] Macrophage related diseases that are treated by the
invention include, but are not limited to, cancer, autoimmune
disease, macrophage activation syndrome, atherosclerosis, diabetes
mellitus, Kawasaki disease, asthma, hemophagocytic
lymphohistiocytosis, sarcoidosis, periodontitis, Whipple's disease,
pulmonary alveolar proteinosis, macrophage related pulmonary
disease, Leishmaniasis, obesity complications, hemodialysis related
inflammation, microbial infection, viral infection, inflammation,
and complications thereof. The cancer can be, without limitation,
adrenal cortical cancer, anal cancer, aplastic anemia, bile duct
cancer, bladder cancer, bone cancer, bone metastasis, central
nervous system (CNS) cancers, peripheral nervous system (PNS)
cancers, breast cancer, Castleman's Disease, cervical cancer,
childhood Non-Hodgkin's lymphoma, colon and rectum cancer,
endometrial cancer, esophagus cancer, Ewing's family of tumors
(e.g., Ewing's sarcoma), eye cancer, gallbladder cancer,
gastrointestinal carcinoid tumors, gastrointestinal stromal tumors,
gestational trophoblastic disease, hairy cell leukemia, Hodgkin's
disease, Kaposi's sarcoma, kidney cancer, laryngeal and
hypopharyngeal cancer, acute lymphocytic leukemia, acute myeloid
leukemia, children's leukemia, chronic lymphocytic leukemia,
chronic myeloid leukemia, liver cancer, lung cancer, lung carcinoid
tumors, Non-Hodgkin's lymphoma, male breast cancer, malignant
mesothelioma, multiple myeloma, myelodysplastic syndrome,
myeloproliferative disorders, nasal cavity and paranasal cancer,
nasopharyngeal cancer, neuroblastoma, oral cavity and oropharyngeal
cancer, osteosarcoma, ovarian cancer, pancreatic cancer, penile
cancer, pituitary tumor, prostate cancer, retinoblastoma,
rhabdomyosarcoma, salivary gland cancer, sarcoma (adult soft tissue
cancer), melanoma skin cancer, non-melanoma skin cancer, stomach
cancer, testicular cancer, thymus cancer, thyroid cancer, uterine
cancer (e.g., uterine sarcoma), vaginal cancer, vulvar cancer, or
Waldenstrom's macroglobulinemia.
[0010] Autoimmune diseases treated by the methods of the invention
include, without limitation, acute disseminated encephalomyelitis,
Addison's disease, alopecia areata, ankylosing spondylitis,
antiphospholipid antibody syndrome, autoimmune hemolytic anemia,
autoimmune hepatitis, Bullous pemphigoid, Celiac disease, Crohn's
disease, dermatomyositis, diabetes mellitus type 1, Goodpasture's
syndrome, Graves' disease, Guillain-Barre syndrome, Hashimoto's
disease, Idiopathic thrombocytopenic purpura, Lupus erythematosus,
Mixed Connective Tissue Disease, multiple sclerosis, myasthenia
gravis, narcolepsy, Pemphigus vulgaris, Pernicious anemia,
Polymyositis, Primary biliary cirrhosis, Rheumatoid arthritis,
Sjogren's syndrome, Temporal arteritis, Ulcerative Colitis,
Vasculitis, or Wegener's granulomatosis.
[0011] Various disease complications treated by the methods of the
invention comprise one or more of retinopathy, neuropathy, foot
problems, gastroparesis, skin complications, bacterial infections,
fungal infections, itching, dermopathy, necrobiosis lipoidica
diabeticorum, diabetic blisters, eruptive xanthomatosis,
nephropathy, hypertension, coronary heart disease, stroke,
rheumatic heart disease, myocardial infraction, metabolic syndrome,
ischemic cardiac disease, coronary artery disease, cerebrovascular
disease, vascular dementia, preeclampsia, heart disease, stroke,
atherogenesis, thrombogenesis, carotid, coronary vascular disease,
transplant-related complications, acute atheroma, metabolic
syndrome, tobacco-related disease, liver complications,
AIDS-related complications, or inflammatory neurological
diseases.
[0012] In one aspect, the present invention provides a method of
treating a macrophage related disease comprising administering to a
subject in need thereof an effective amount of a pharmaceutical
composition comprising an oxidative agent, wherein the macrophage
related disease is a neurological disease. In another aspect, the
present invention provides a method of treating a neurological
disease comprising administering to a subject in need thereof an
effective amount of a pharmaceutical composition comprising an
oxidative agent. In another aspect, the present invention provides
a method of treating a neurological disease comprising
administering to a subject in need thereof an effective amount of a
pharmaceutical composition comprising chlorite or a
chlorite-containing agent, optionally in combination with another
therapeutic agent or intervention used for treating the
neurological disorder.
[0013] In another aspect, the present invention provides a method
of treating a symptom of a neurological disease comprising
administering to a subject in need thereof an effective amount of a
pharmaceutical composition comprising chlorite or a
chlorite-containing agent in combination with another therapeutic
agent or intervention used for treating the symptom. Symptoms
treated by the methods of the invention include but are not limited
to motor impairment, cognitive impairment, dementia, pain,
disorganization, sleep disorder, infection, depression, anxiety,
pneumonia, compulsive behavior, injuries, dysphagia, constipation,
incontinence, akinesia, restless legs syndrome, tremor, agitation,
sensory impairment, visual impairment, smell impairment, hearing
impairment, or osteoporosis.
[0014] The methods the invention can be used to treat any number of
neurological diseases, disorders, or symptoms of any thereof. In
some embodiments, the neurological disease is amyotrophic lateral
sclerosis (ALS). For ALS, the second therapeutic agent or
intervention includes but is not limited to riluzole, KNS-760704,
minocycline, RNAi targeting SOD1 gene, physical therapy,
insulin-like growth factor-1(IGF1), brain-derived neurotrophic
factor (BDNF), nerve growth factor (NGF) like factors,
5-hydroxytryptophan (5-HTP), tyrosine, levodopa (L-DOPA),
tryptophan, cysteine, exercise, ventilatory assistance or
communication assistance. For ALS, the second therapeutic agent or
intervention can also comprise 2-PMPA, adenosine, anisomycin,
apocynin, apomorphine, arimoclomol, aspergillin, BMP-7,
carboxyfullerenes, ceftriaxone, celastrol, geldanamycin, celecoxib
(Celebrex.RTM.), cyclooxygenase 2, CGP 3466B, chlorpromazine,
clioquinol, clozapine, ciliary neurotrophic factor (CTNF),
colchicine, colivelin, copaxone, copper chelators, lipoic acid,
coenzyme Q10 (CoQ10), creatine, curcumin, cytotoxic T-lymphocyte
antigen 4 antibody fusions (CTLA4-Ig), cycloheximide, cobra venom
factor (CVF), cycloserine, cyclosporin, d-penicillamine,
JAK3/Dapsone, Dapsone/Gusperimus/JAK3 Cocktail,
Diethyldithiocarbamate DDC, desferoxamine, desipramine,
.alpha.-difluoromethylornithine (DFMO), dietary restriction,
dihydrotestosterone, 5,5-dimethyl-pyrroline N-oxide (DMPO),
excitatory amino acid transporter 2 (EAAT2),
erythro-9-[3-(2-hydroxynonyl)] adenine (EHNA), emetine, estradiol
benzoate, exercise, FK-506, fluorouracil, glial cell line-derived
neurotrophic factor (GDNF), decreased spinal copper levels,
genistein, glutamate receptor 3 (GLUR3) antisense, hepatocyte
growth factor (HGF), hNT neurons, anti-oxidant SOD1 protein, human
umbilical cord blood mononuclear cells, hydroxyurea,
interleukin-1beta-converting enzyme (ICE) inhibition, IGF-1 or
isoforms thereof, intravenous immunoglobulin (IVIG), indomethacin,
hydroquinone hydrochloride derivative of 17-AAG (IPI-504), iron
porphyrin, ivermectin (22,23-dihydroavermectin
B1a+22,23-dihydroavermectin B1b), L-acetyl-carnitine, lactacystin,
leflunomide, lentiviral RNAi SOD1 gene silencing, leukemia
inhibitory factor (LIF), lithium, lyophilized red wine extract,
magnesium supplementation, melatonin, memantine, metalloporphyrins
(MnTE-Py-P (AEOL10113 and AEOL10150)), metallothioneins, metformin,
methotrexate, mechano growth factor (MGF; IGF-I Ec peptide; mIGF-1
isoform), minocycline, minocycline/creatine,
minocycline/riluzole/nimodipine cocktail, mithramycin,
1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and/or
3-nitropropionic acid (3NP), N-acetyl-L-cysteine, N-acetylcysteine,
2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo[f]quinoxaline-2,3-dione
(NBQX), nordihydroguaiaretic acid (NDGA), neurofilament heavy
(NF-H) protein, neurofilament light (NF-L) protein, nimesulide,
nitric oxide synthase inhibitors, 17 beta-estradiol, p75
neurotrophin receptor, p75 neurotrophin agonist, p75 neurotrophin
antisense, parvalbumin, sodium phenyl butyrate (PBA), peripheral
axotomy, phosphatidyl choline-bound Cu/Zn SOD, pioglitazone,
polyamine-modified catalase, porphyrin, prednisolone, progesterone,
puromycin, putrescine-modified catalase (PUT-CAT), quinacrine, R(+)
pramipexole, radicicol, rasagiline, resveratrol/red wine extract,
riluzole, ritonavir, anti-myostatin mAb, vascular endothelial
growth factor (VEGF), RNAi targeting human SOD1 gene, rofecoxib,
rolipram, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid
antagonist RPR 119990, 5-hydroxytryptophan (5-HTP), sodium
valproate, stem cells, sulindac, tamoxifen, propargylamine TCH346,
tepoxalin, testosterone, thalidomide, trehalose, trichostatin A,
trientine/ascorbate, vincristine, vitamin E/riluzole/gabapentin,
Janus kinase 3 (JAK3) inhibitor such as WHI-P131, bone marrow
transplant, zileuton, zinc sulfate, or a noncompetitive AMPA
antagonist such as ZK 187638. The invention further contemplates
combination therapy for ALS comprising administering chlorite
and/or a chlorite containing agent and any appropriate combination
of other agents or interventions.
[0015] In some embodiments, the neurological disease treated by the
methods of the invention is Parkinson's disease (PD). For
Parkinson's disease, the second therapeutic agent or intervention
includes but is not limited to antioxidant, immunosuppressive
calcineurin inhibitor, NOS inhibitor, sigma-1 modulator, AMPA
antagonist, Ca2+ channel blocker, estrogen agonist, MAO-B
inhibitor, selegiline, rasagiline, kinase inhibitor, mitochondrial
modulator or enhancer, alpha synuclein modulator, glycoprotein
IIb/IIIa antagonist, erythropoietin, astaxanthin, boswellia,
caffeine, curcumin, E vitamins, tocotrienol, flavonoid, naringenin,
huperzine, ubiquinol, levodopa (L-DOPA), levodopa-carbidopa,
co-beneldopa, a slow-release levodopa mono-therapy or combination
formulation, amantadine, benztropine, procyclidine,
trihexyphenidyl, a COMT inhibitor, tolcapone, entacapone, a
dopamine agonist, bromocriptine, pergolide, pramipexole,
ropinirole, cabergoline, apomorphine, lisuride, surgery, deep brain
stimulation, neurorehabilitation, or exercise. The invention
further contemplates combination therapy for Parkinson's disease
comprising administering chlorite and/or a chlorite containing
agent and any appropriate combination of other agents or
interventions.
[0016] In some embodiments, the neurological disease treated by the
methods of the invention is Alzheimer's disease (AD). For
Alzheimer's disease, the second therapeutic agent or intervention
includes but is not limited to an acetylcholinesterase inhibitor,
donepezil, rivastigmine, galantamine, THA, an NMDA receptor
antagonist, memantine, vitamin E, vitamin D, progesterone, or a
behavioral, emotional, cognitive, or stimulation-oriented
intervention. The invention further contemplates combination
therapy for Alzheimer's disease comprising administering chlorite
and/or a chlorite containing agent and any appropriate combination
of other agents or interventions.
[0017] In some embodiments, the oxidative agent of the invention is
chlorite or a chlorite-based derivative or functional product. In
some embodiments, the oxidative agent is Tetrachlorodecaoxygen
(TCDO). In some embodiments, the oxidative agent is chloramine-T or
a hydrate thereof or taurine chloramine In some embodiments, the
oxidative agent is WF10. In some embodiments, the oxidative agent
is heme oxygenase 1 (HO -1) or a functional product thereof. In
some embodiments, the oxidative agent is potassium nitrate (KNO3),
hypochlorite, a hypohalite compound, fluorine (F), chlorine (Cl),
bromine (Br), iodine (I), astatine (At), chlorite, chlorate,
perchlorate, a halogen-containing compound, a permanganate salt,
ammonium cerium(IV) nitrate, a Cerium(IV) compound, a hexavalent
chromium compound, chromic acid, dichromic acid, chromium trioxide,
pyridinium chlorochromate (PCC), a chromate/dichromate compounds, a
peroxide, Tollens' reagent, a sulfoxide, persulfuric acid, ozone,
osmium tetroxide (OsO4), nitric acid, nitrous oxide (N2O), or a
derivative of any thereof.
[0018] In some embodiments, the immunomodulatory agent is an
immunosuppressive agent. The immunomodulatory agent can also be an
immunostimulator. In some embodiments, the immunomodulatory agent
is a glucocorticoid, hydrocortisone (cortisol), cortisone acetate,
prednisone, prednisolone, methylprednisolone, dexamethasone,
betamethasone, triamcinolone, beclometasone, fludrocortisone
acetate, deoxycorticosterone acetate (doca) aldosterone, a
non-glucocorticoid steroid, a cytostatic agent, an alkylating
agent, nitrogen mustard (cyclophosphamide), nitrosourea, a platinum
compound, an antimetabolite, a purine analog, azathioprine,
mercaptopurine, mycophenolic acid, a pyrimidine synthesis
inhibitor, leflunomide, teriflunomide, a folic acid analog,
methotrexate, a cytotoxic antibiotic, dactinomycin, anthracyclines,
mitomycin C, bleomycin, mithramycin, an antibody or fusion thereof,
anti-thymocyte globulin, anti-lymphocyte globulin, an anti-IL-2
receptor antibody, an anti-CD3 antibody, OKT3 (muromonab),
otelixizumab, teplizumab, visilizumab, an anti-CD4 antibody,
clenoliximab, keliximab, zanolimumab, an anti-CD11a antibody,
efalizumab, an anti-CD18 antibody, erlizumab, rovelizumab, an
anti-CD20 antibody, afutuzumab, ocrelizumab, ofatumumab,
pascolizumab, rituximab, an anti-CD23 antibody, lumiliximab, an
anti-CD40 antibody, teneliximab, toralizumab, an anti-CD40L
antibody, ruplizumab, an anti-CD62L antibody, aselizumab, an
anti-CD80 antibody, galiximab, an anti-CD147 antibody, gavilimomab,
a B-Lymphocyte stimulator (BLyS) inhibiting antibody, belimumab, an
CTLA4-Ig fusion protein, abatacept, belatacept, an anti-CTLA4
antibody, ipilimumab, tremelimumab, an anti-eotaxin 1 antibody,
bertilimumab, an anti-.alpha.4-integrin antibody, natalizumab, an
anti-IL-6R antibody, tocilizumab, an anti-LFA-1 antibody,
odulimomab, an anti-CD25 antibody, basiliximab, daclizumab,
inolimomab, an anti-CD5 antibody, zolimomab, an anti-CD2 antibody,
siplizumab, nerelimomab, faralimomab, atlizumab, atorolimumab,
cedelizumab, dorlimomab aritox, dorlixizumab, fontolizumab,
gantenerumab, gomiliximab, lebrilizumab, maslimomab, morolimumab,
pexelizumab, reslizumab, rovelizumab, talizumab, telimomab aritox,
vapaliximab, vepalimomab, aflibercept, alefacept, rilonacept, an
immunophilin modulating agent, rapamycin, a calcineurin inhibitor,
tacrolimus, ciclosporin (cyclosporin), pimecrolimus, abetimus,
gusperimus, ridaforolimus, everolimus, temsirolimus, zotarolimus, a
TNF inhibitor, infliximab, adalimumab, certolizumab pegol,
golimumab, etanercept, thalidomide, lenalidomide, pentoxifylline,
bupropion, curcumin, catechin, an IL-1 receptor antagonist,
anakinra, an anti-IL-5 antibody, mepolizumab, an IgE inhibitor,
omalizumab, talizumab, an IL12 inhibitor, an IL23 inhibitor,
ustekinumab, an opiod, an IMPDH inhibitor, mycophenolic acid,
myriocin, fingolimod, an NF-.kappa.B inhibitor, raloxifene,
drotrecogin alfa, denosumab, an NF-.kappa.B signaling cascade
inhibitor, disulfiram, olmesartan, dithiocarbamate, a proteasome
inhibitor, bortezomib, MG132, Pro1, NPI-0052, curcumin, genistein,
resveratrol, parthenolide, thalidomide, lenalidomide, flavopiridol,
non-steroidal anti-inflammatory drugs (NSAIDs), arsenic trioxide,
dehydroxymethylepoxyquinomycin (DHMEQ),
I3C(indole-3-carbinol)/DIM(di-indolmethane) (I3C/DIM), Bay 11-7082,
luteolin, cell permeable peptide SN-50, I.kappa.B.alpha.-super
repressor overexpression, NF.kappa.B decoy oligodeoxynucleotide
(ODN), or a derivative or analog of any thereof.
[0019] The pharmaceutical compositions of the invention can be
administered intravenously. In some embodiments, the subject
treated using the methods of the invention is a mammal. In some
embodiments, the mammal is a human.
[0020] In some embodiments, modulating of macrophage activation or
accumulation according to the invention comprises modulating one or
more molecules involved in one or more cellular pathways selected
from the group consisting of NFkB, toll-like receptor (TLR)-2,
TLR-4, Tie-2/Ang-2, TRIF/TBK1/IRF3, NFAT, and hypoxia-induced
pathways, lipopolysaccharide (LPS), prostaglandin E2 (PGE2),
interferon (IFN)-.alpha., IFN-.beta., IFN-.gamma., interleukin
(IL)-1, IL-4, IL-6, sIL1Ra, IL-8, IL-10, IL-12, IL-12p40, IL-13,
IP10, MHCII, TNF-.alpha., macrophage inflammatory protein 1 alpha
(MIP1-.alpha.), IFN-gamma-inducing factor (IGIF),
macrophage-stimulating protein (MSP), inter-cellular adhesion
molecule 1(ICAM-1), colony stimulating factor 1 (CSF-1R),
L-arginine, nitric oxide signaling pathways, and macrophage
migration inhibitory factor (MIF). In some embodiments, the
modulating of macrophage activation with the agent has effect on
one or more molecules selected from the group consisting of TLR-2,
TLR-4, mkp-1, COX-2, SOCS-3, Fc.gamma.R1, IFN-.alpha., IFN-.beta.,
IL-4, IL-6, IL-1Ra, IGIF, IL-.beta., MHCI, MHCII IAA, MHCII IAB,
MHCII IEB, IP10, IL-10, cathepsin H, lysozyme, CathB, stk,
TNF-.alpha., IL-12p35, IL-12p40, MIP-1.alpha., ICAM-1, INOS, mig,
Cat-2, CIITA, ICSBP, CathL, CSF1R, GM-CSF, IRF1, IRF-2, c-fos,
VEGF, IL-8, bFGF, CSF-1, EGF, MMP-2, MMP-7, MMP-9, MMP-12, EMAPII,
endothelin 2, HIF-1, HIF-2, CXCL8, TGF.beta., PGE2, and MDF.
[0021] In some embodiments, the methods of the invention further
comprise administering a second therapeutic agent sequentially or
simultaneously.
[0022] In the case of proliferative diseases such as cancer, the
second therapeutic agent can comprise a chemotherapeutic,
immunotherapeutic, or radiotherapeutic agent. Such agents can be
selected from the group consisting of alkylating agents,
antimetabolites, natural antineoplastic agents, hormonal
antineoplastic agents, angiogenesis inhibitors, differentiating
reagents, RNA inhibitors, antibodies or immunotherapeutic agents,
gene therapy agents, small molecule enzymatic inhibitors,
biological response modifiers, and anti-metastatic agents.
[0023] In the case of autoimmune diseases, the second therapeutic
agent can comprise an immunosuppressant or anti-inflammatory agent.
Such agents can be selected from the group consisting of alkylating
agent, an antimetabolite, a cytotoxic antibiotic, a folic acid
analog, a purine analog, an antibody, a TNF binding protein, an
interferon, an opioid, a mycophenolate, a calcineurin inhibitor, or
an analog thereof.
[0024] In some embodiments, the second therapeutic agent comprises
a therapeutic agent for treating type II diabetes or related
complications. Such agents include without limitation a biguanide,
metformin, a thiazolidinedione, ciglitazone, pioglitazone,
troglitazone, rosiglitazone, a dipeptidyl-peptidase-4 inhibitors,
vildagliptin, sitagliptin, a glucagonlike peptide-1 ("GLP-1")
receptor agonist, exanatide, a GLP-1 mimetic, a PPAR gamma agonist
or partial agonist, a dual PPAR alpha-PPAR gamma agonist or partial
agonist, a dual PPAR delta-PPAR gamma agonist or partial agonist, a
pan PPAR agonist or partial agonist, dehydroepiandrosterone or its
conjugated sulphate ester, an antiglucocorticoid, a TNF-alpha
inhibitor, an alpha-glucosidase inhibitor, acarbose, miglitol,
voglibose, a sulfonylurea, chlorpropamide, tolbutamide,
acetohexamide, tolazamide, glyburide, gliclazide, glynase,
glimepiride, glipizide, pramlintide, an insulin secretogogue,
repaglinide, gliquidone, nateglinide, insulin, an insulin mimetic,
a glucagon receptor antagonist, gastric inhibitory peptide ("GIP"),
a GIP mimetic, ketotifen fumarate or cromolyn.
[0025] In some embodiments, the second therapeutic agent comprises
a therapeutic agent used for treating a vascular disease and/or
related disorders. Such agents include without limitation an
endothelin receptor antagonist, bosentan, darusentan, enrasentan,
tezosentan, atrasentan, ambrisentan sitaxsentan, a smooth muscle
relaxant, a PDE5 inhibitor, minoxidil, an angiotensin converting
enzyme (ACE) inhibitor, captopril, enalapril, lisinopril,
fosinopril, perindopril, quinapril, trandolapril, benazepril,
ramipril, an angiotensin II receptor blocker, irbesartan, losartan,
valsartan, eprosartan, olmesartan, candesartan, telmisartan, a beta
blocker, atenolol, metoprolol, nadolol, bisoprolol, pindolol,
acebutolol, betaxolol, propranolol, a diuretic, thiazide,
hydrochlorothiazide, furosemide, torsemide, metolazone, a calcium
channel blocker, amlodipine, felodipine, nisoldipine, nifedipine,
verapamil, diltiazem, an alpha receptor blocker, doxazosin,
terazosin, alfuzosin, tamsulosin, a central alpha agonist,
clonidine, a statin, atovastatin, fluvastatin, lovastatin,
pravastatin, rosuvastatin calcium, simvastatin, nicotinic acid, an
agent that stimulate PPAR alpha, a fibrate, gemfibrozil,
fenofibrate, bezafibrate, ciprofibrate, a bile acid sequestrant,
cholestyramine, colestipol, a cholesterol absorption inhibitor, a
COX-1 inhibitor, aspirin, an NSAID, or a COX-2 inhibitor.
[0026] In some embodiments, the second therapeutic agent comprises
a therapeutic agent used for treating obesity and/or related
disorders. Such agents include without limitation
phenylpropanolamine, phenteramine, diethylpropion, mazindol,
fenfluramine, dexfenfluramine, phentiramine, a beta-3 adrenoceptor
agonist agent, sibutramine, a gastrointestinal lipase inhibitor,
orlistat, a leptin, a cannabinoid-1 ("CB-1") receptor antagonist,
rimonabant, a PPAR delta agonist or partial agonist, a dual PPAR
alpha-PPAR delta agonist or partial agonist, a dual PPAR delta-PPAR
gamma agonist or partial agonist, a pan PPAR agonist or partial
agonist, neuropeptide Y, enterostatin, cholecytokinin, bombesin,
amylin, a histamine H3 receptor, a dopamine D2 receptor, melanocyte
stimulating hormone, corticotrophin releasing factor, galanin,
gamma amino butyric acid (GABA), ketotifen fumarate or
cromolyn.
[0027] In some embodiments, the second therapeutic agent comprises
a therapeutic agent used for treating atherosclerosis. Such agents
include without limitation an antihyperlipidemic agent, a plasma
HDL-raising agent, an antihypercholesterolemic agent, a cholesterol
biosynthesis inhibitor, an hydroxymethylglutaryl (HMG) CoA
reductase inhibitor, a statin, lovastatin, simvastatin,
pravastatin, fluvastatin, and atorvastatin, an HMG-CoA synthase
inhibitor, a squalene epoxidase inhibitor, a squalene synthetase
inhibitor, a squalene synthase inhibitor, an acyl-coenzyme A
cholesterol acyltransferase (ACAT) inhibitor, melinamide, probucol,
nicotinic acid and salts thereof, niacinamide, a cholesterol
absorption inhibitor, beta-sitosterol, a bile acid sequestrant
anion exchange resin, cholestyramine, colestipol, a
dialkylaminoalkyl derivative of a cross-linked dextran, an LDL
receptor inducer, a fibrate, clofibrate, bezafibrate, fenofibrate,
gemfibrizol, vitamin B6 (pyridoxine) and pharmaceutically
acceptable salts thereof, vitamin B6 HCl salt, vitamin B12
(cyanocobalamin), vitamin B3 (nicotinic acid or niacinamide), an
anti-oxidant vitamin, vitamin C, vitamin E, beta carotene, a
beta-blocker, an angiotensin II antagonist, an angiotensin
converting enzyme inhibitor, a PPAR alpha agonist or partial
agonist, a PPAR delta agonist or partial agonist, a PPAR gamma
agonist or partial agonist, a dual PPAR alpha-PPAR delta agonist or
partial agonist, a dual PPAR alpha-PPAR gamma agonist or partial
agonist, a dual PPAR delta-PPAR gamma agonist or partial agonist, a
pan PPAR agonist or partial agonist, a platelet aggregation
inhibitor, a fibrinogen receptor antagonists, a glycoprotein
IIb/IIIa fibrinogen receptor antagonist, or aspirin.
[0028] In some embodiments, the second therapeutic agent comprises
a therapeutic agent used for treating hyperlipidemia. Such agents
include without limitation a statin, atorvastatin, fluvastatin,
lovastatin, pravastatin, simvastatin, a CETP inhibitor,
torcetrapib, a cholesterol absorption inhibitor, ezetimibe, a PPAR
alpha agonist or partial agonist, a PPAR delta agonist or partial
agonist, a dual PPAR alpha-PPAR delta agonist or partial agonist, a
dual PPAR alpha-PPAR gamma agonist or partial agonist, a dual PPAR
delta-PPAR gamma agonist or partial agonist, a pan PPAR agonist or
partial agonist, a fenofibric acid derivative, gemfibrozil,
clofibrate, fenofibrate, bezafibrate, a bile acid-binding resin,
colestipol, cholestyramine, nicotinic acid and salts thereof,
probucol, betacarotene, vitamin E, or vitamin C.
[0029] In some embodiments the invention provides a method of
treating Type II diabetes or related complications comprising
administering to a subject in need thereof an effective amount of a
pharmaceutical composition comprising chlorite or a
chlorite-containing agent.
[0030] In some embodiments the invention provides a method of
treating a disease associated with migration of monocytes or
activated macrophages, the method comprising administering a
therapeutically-effective amount of an oxidative agent to a subject
in need thereof, where the oxidative agent is selected from the
group consisting of non-halogen activated-oxygen compounds,
non-oxygen activated-halogen compounds, activated-halogen
activated-oxygen compounds, and N-halo compounds. In some
embodiments, the disease is characterized by elevated CD16
expression levels in CD14+ cells. In some embodiments, the
oxidative agent is selected from sodium chlorite,
1,3-dichloro-5,5-dimethylhydantoin and chloramine-T. In some
embodiments, the disease is characterized by migration of PBMCs in
response to chemoattractant. In some embodiments, the oxidative
agent is sodium chlorite.
[0031] In some embodiments, the invention provides a method of
treating a disease associated with excess production of sCD14
and/or sCD163 by activated macrophages, the method comprising
administering a therapeutically-effective amount of an oxidative
agent to a subject in need thereof, where the oxidative agent is
selected from the group consisting of non-halogen activated-oxygen
compounds, non-oxygen activated-halogen compounds,
activated-halogen activated-oxygen compounds, and N-halo compounds.
In some embodiments, the oxidative agent is sodium chlorite.
[0032] In some embodiments, the method of diagnosing a macrophage
related disease in a subject comprising measuring the level of a
biomarker in the subject and correlating the measured level of
biomarker to normal and diseased levels of said biomarker, where
the biomarker is selected from CD16 expression in CD14+ cells,
sCD14, sCD163, expression of chemoattractants by macrophages, and
combinations thereof.
[0033] In some embodiments, the invention provides a method of
determining efficacy of treatment with an oxidative agent for a
macrophage related disease in a subject comprising the steps of: i)
initiating treatment with an oxidative agent; ii) measuring the
level of a biomarker in the subject; and iii) correlating the
measured level of biomarker to normal and diseased levels of the
biomarker and/or levels of biomarker in said subject prior to
treatment; where the biomarker is selected from CD16 expression in
CD14+ cells, sCD14, sCD163, expression of chemoattractants by
macrophages, and combinations thereof. In some embodiments,
oxidative treatment is sodium chlorite.
[0034] In some embodiments, the invention provides a sodium
chlorite compound, where the compound is a crystalline solid of
greater than 95% purity. In some embodiments, the compound is a
crystalline solid of greater than 99% purity. In some embodiments,
the compound has an x-ray powder diffraction pattern with peaks
expressed in degrees 20 at about 21, 30, 31, 32, 34, and 39. In
some embodiments, the invention provides a pharmaceutical
composition comprising one or more pharmaceutical excipients and
the sodium chlorite compound. In some embodiments, the invention
provides a pharmaceutical composition comprising: (a) the sodium
chlorite compound; (b) a pH adjusting agent; and (c) a
pharmaceutically acceptable excipient or carrier, wherein said
composition is a liquid that exhibits 25% less pH drift compared to
an identical composition without said pH adjusting agent. In some
embodiments, the pH adjusting agent is sodium phosphate
dibasic.
[0035] In another aspect, the present invention provides a kit
comprising: (a) one or more unit dose forms comprising an effective
amount of a pharmaceutical composition comprising an oxidative
agent and/or an immunomodulatory agent; and (b) one or more of
packaging and instructions for use to treat a macrophage related
disease. In some embodiments, the oxidative agent is chlorite or a
chlorite-based compound. In some embodiments, the unit dose form is
ready for administration to a subject. In some embodiments, the
unit dose form is to be diluted prior to administration to a
subject.
INCORPORATION BY REFERENCE
[0036] All publications and patent applications mentioned in this
specification are herein incorporated by reference to the same
extent as if each individual publication or patent application was
specifically and individually indicated to be incorporated by
reference.
BRIEF DESCRIPTION OF THE DRAWINGS
[0037] The novel features of the invention are set forth with
particularity in the appended claims. A better understanding of the
features and advantages of the present invention will be obtained
by reference to the following detailed description that sets forth
illustrative embodiments, in which the principles of the invention
are utilized, and the accompanying drawings of which:
[0038] FIG. 1 shows median CD14CD16 cell surface expression levels
after exposure of five samples of normal PBMCs to WF10 or chlorite
at various concentrations for three days.
[0039] FIG. 2 shows that treatment of cells from five amyotrophic
lateral sclerosis (ALS) blood samples with chlorite (striped bars)
causes down regulation of CD16 expression on CD14+ cells.
[0040] FIG. 3 shows osteopontin secretion by monocytes regulated by
chlorite. WF10 (OPN-W) and chlorite (OPN-C) inhibit osteopontin
secretion by monocytes to a similar extent.
[0041] FIG. 4 shows cytokine secretion by ALS peripheral blood
mononuclear cells (PBMCs) regulated by chlorite. Both WF10 and
chlorite inhibit MCP-1, osteopontin (OPN), and MMP-9 secretion by
PBMCs from amyotrophic lateral sclerosis (ALS) blood to a similar
extent. WF10 and chlorite do not have a significant effect on IL-6
secretion by ALS PBMCs as compared to the control.
[0042] FIG. 5 shows a Thermo-Gravimetric Analysis (TGA) of a sample
of sodium chlorite purified according to the invention. The
thermogram shows the loss of a total of 40.0% weight from ambient
temperature to about 160.degree. C.
[0043] FIG. 6 shows an X-ray powder diffraction (XRPD) pattern for
a sample of sodium chlorite purified according to the invention.
The XRPD pattern indicates that the material is crystalline.
[0044] FIG. 7 shows various pathways for modulating macrophage
related disorders.
[0045] FIG. 8 shows the effect of various compounds on CD16
expression in CD14+ cells as well as monocyte toxicity.
[0046] FIGS. 9, 10, and 11 show the experimental design for Example
8, the effect of ALS patient macrophage supernatant on blood
monocyte migration, and the effect of sodium chlorite on blood
monocyte migration.
[0047] FIG. 12 shows the effect of sodium chlorite on ALS patient
monocytes in blocking production of sCD14 and sCD163.
DETAILED DESCRIPTION OF THE INVENTION
[0048] In one aspect, the present invention provides a method of
treating a macrophage related diseases and conditions comprising
administering to a subject in need thereof an effective amount of
an oxidative agent. Macrophage related diseases and conditions
include those which result directly from aberrant macrophage
action, and also include any disease in which macrophages play a
role, e.g., by inducing inflammation. In another aspect, the
present invention provides a method of modulating macrophage
accumulation or activation comprising administering to a subject in
need thereof an effective amount of an oxidative agent. In some
embodiments, macrophage activation is enhanced with the oxidative
agent of the present invention. In other embodiments, macrophage
activation is reduced or inhibited with the oxidative agent of the
present invention. In still another aspect, the present invention
provides a method of treating a macrophage related disease
comprising administering to a subject in need thereof an effective
amount of an immunomodulating agent, e.g., an immunosuppressive
agent. Modulating macrophage function according to the present
invention can have direct therapeutic effect, e.g., by reducing
harmful inflammation. In addition, other therapeutic agents may
work more effectively in the non-inflamed environment.
[0049] In some embodiments, the oxidative agent for use with the
present invention comprises chlorite. In some embodiments, a
chlorite formulation comprises chlorite, a buffer, e.g., monosodium
phosphate buffer or disodium phosphate buffer, and a pharmaceutical
excipient. In some embodiments, the oxidative agent is WF10,
chloramine-T or taurine chloramine
[0050] Immunomodulators, eg, immunosuppressants for use with the
invention include without limitation cyclophosphamide,
dexamethasone, cyclosporine, azathioprine, methotrexate, FK-506 and
rapamycin. Antibodies against cell surface receptors which modulate
the immune response can also be used. For instance, some antibodies
and soluble receptor ligands that block ligand binding to cellular
receptors on B cells, T cells, NK cells, and macrophages can be
used to downregulate the macrophage response directly or
indirectly.
[0051] Macrophage-related diseases that can be treated by the
methods of the invention include but are not limited to cancer,
autoimmune diseases such as rheumatoid arthritis and multiple
sclerosis, metabolic disorder, diabetes, peripheral vascular and
cardiovascular disorders, infection, inflammation, and adverse
effects of obesity. In some embodiments, oxidative agents or
immunomodulators are used to treat a disease that is related to
macrophages or monocytes. In some embodiments, the macrophage
related disease is an inflammatory disease. In some embodiments,
the macrophage related disease involves granuloma formation. In
some embodiments, the macrophage related disease is a cancer. In
some embodiments, the oxidative agent or immunomodulating agent
modulates a pathway involved in macrophage activation. The
macrophage activation related cellular pathways that can be
modulated by the methods of the present invention include but are
not limited to TLR4, CAT2, ICSBP, IFNR-I, IFNR-II, IRF1, IRF2,
Raf-1, MEK1, MEK2, ERK1, ERK2, p38, MAPKK4, MAPKK6, PKC, JAK1,
JAK2, STAT1, STAT3, Elk1, JNK/SAPK, AP1, Pu1, NFkB, NFAT, iNOS,
USF1, ISGF3, SP1, Bc16, ATF2, c-Jun, and COX-2. Molecules important
to macrophage activation, function or effects that can be
modulated, either directly or indirectly, by the methods of the
present invention include but are not limited to TLR-2, TLR-4,
mkp-1, COX-2, SOCS-3, Fc.gamma.R1, IFN-.alpha., IFN-.beta., IL-4,
IL-6, IL-1Ra, IGIF, IL-1.beta., MHCI, MHCII IAA, MHCII IAB, MHCII
IEB, IP10, IL-10, cathepsin H, lysozyme, CathB, stk, TNF-.alpha.,
IL-12p35, IL-12p40, MIP-1.alpha., ICAM-1, INOS, mig, Cat-2, CIITA,
ICSBP, CathL, CSF1R, GM-CSF, IRF1, IRF-2, c-fos, VEGF, IL-8, bFGF,
CSF-1, EGF, MMP-2, MMP-7, MMP-9, MMP-12, EMAPII, endothelin 2,
HIF-1, HIF-2, CXCL8, TGF.beta., PGE2, and MDF.
A. Oxidative Agents
[0052] In one aspect, the present invention provides a method of
treating a macrophage related disease comprising administering to a
subject in need thereof an effective amount of an oxidative agent.
In another aspect, the present invention provides a method of
modulating macrophage accumulation or activation comprising
administering to a subject in need thereof an effective amount of
an oxidative agent.
[0053] I. Chlorite and Other Oxidative Agents
[0054] Substances that have the ability to oxidize other substances
are typically referred to as oxidative and are known as oxidizing
agents, oxidants, or oxidizers, which are used interchangeably
herein. An oxidizing agent (also called an oxidant, oxidizer) can
be defined as either: a chemical compound that readily transfers
oxygen atoms, or a substance that gains electrons in a redox
chemical reaction. In both cases, the oxidizing agent becomes
reduced in the process. Various common oxidizers contain oxygen
(e.g., KClO.sub.4) and can be considered as storage forms of
oxygen. Alternatively, the term "oxidizing agent" also includes any
time where formal charge is increased (losing electrons), and
applies to substances that contain no oxygen, typically halogens
comprising fluorine, (F); chlorine, (Cl); bromine, (Br); iodine,
(I); and astatine, (At), and substances rich in these elements.
[0055] Common oxidizing or oxidative agents that can be used in the
methods of the present invention include but are not limited to
potassium nitrate (KNO.sub.3), hypochlorite and other hypohalite
compounds, iodine and other halogens, chlorite, chlorate,
perchlorate, and other analogous halogen compounds, permanganate
salts, ammonium cerium(IV) nitrate and related cerium(IV)
compounds, hexavalent chromium compounds such as chromic and
dichromic acids and chromium trioxide, pyridinium chlorochromate
(PCC), and chromate/dichromate compounds; peroxide compounds,
Tollens' reagent, sulfoxides, persulfuric acid, ozone, osmium
tetroxide (OsO.sub.4), nitric acid, and nitrous oxide (N.sub.2O).
In one embodiment, the oxidative agent is non-toxic to CD14
monocytes at physiologically effective concentrations.
[0056] In one aspect, the oxidative agents of the invention are
compounds that readily transfer oxygen atoms and/or provide compact
storage of oxygen while containing no halogens. As used herein,
such compounds are referred to as non-halogen activated-oxygen
compounds, and include but are not limited to potassium nitrate
(KNO.sub.3), permanganate salts, ammonium cerium(IV) nitrate and
related cerium(IV) compounds, hexavalent chromium compounds such as
chromic and dichromic acids and chromium trioxide, and
chromate/dichromate compounds; peroxide compounds, Tollens'
reagent, ammonium silver nitrate, sulfoxides, persulfuric acid,
ozone, osmium tetroxide (OsO.sub.4), nitric acid, nitrous oxide
(N.sub.2O), hydrogen peroxide, organic peroxides, superoxides, and
ozone.
[0057] In one aspect, the oxidative agents of the invention are
compounds that contain both readily-transferrable oxygen and
halogen atoms, including but not limited to hypochlorite and other
hypohalite compounds, chlorite, chlorate, perchlorate and other
analogous halogen compounds, and pyridinium chlorochromate (PCC).
As used herein, such compounds are referred to as activated-oxygen
activated-halogen compounds.
[0058] Alternatively, the oxidative agent may be a substance that
contains no oxygen, typically halogens comprising fluorine, (F);
chlorine, (Cl); bromine, (Br); iodine, (I); and astatine, (At). As
used herein, such compounds are referred to non-oxygen
activated-halogen compounds.
[0059] In one embodiment, the oxidative agent may be an N-halo
compound. Such compounds include but are not limited to
N-halophthalimide, N-halosuccinimide, N-halosaccharin,
N,N-dihalourethane, N-haloacetanilide,
1,3-dihalo-5,5-dimethylhydantoin, trihaloisocyanuric acid, and
sodium dihaloisocyanurate, where the halogen is selected from
fluorine, chlorine, bromine, and iodine. Preferably, the oxidative
agent may be an N-chloro compound. Such compounds include but are
not limited to N-chlorophthalimide, N-chlorosuccinimide,
N-chlorosaccharin, N,N-dichlorourethane, N-chloroacetanilide,
1,3-dichloro-5,5-dimethylhydantoin, trichloroisocyanuric acid,
sodium dichloroisocyanurate, chloramine-T (including hydrate), and
halazone (4-(N,N-dichlorosulfamoyl)benzoic acid). In one
embodiment, the oxidative agent is selected from the group
consisting of 1,3-dichloro-5,5-dimethylhydantoin and
chloramine-T.
[0060] Many oxidative compounds have demonstrated protective and
anti-inflammatory activities, likely due to induction of endogenous
defense pathways. For example, metabolites of the stress induced
enzyme heme oxygenase 1 (HO-1) such as carbon monoxide (CO) and
biliverdin exert potent anti-inflammatory effects (Otterbein L E et
al. Nat. Med. 6 (2000) 422-428). The catalytic products of HO-1
including the oxidants CO, Fe2+, and biliverdin are capable of
down-regulating inflammatory reactions. Similar cell-protective
properties have been described for the redox-active molecule
thioredoxin (Hirota K. et al. J. Biol. Chem. 274 (1999)
27891-27897). The use of chlorite to treat various diseases and
conditions is described in U.S. Pat. No. 4,725,437; U.S. Pat. No.
4,851,222; McGrath et al., Development of WF10, a novel
macrophage-regulating agent, Curr Opin Investig Drugs, 3(3):365-73
(March 2002); U.S. Pat. No. 6,086,922; US Patent Pub. No.
2005/0181068 (Ser. No. 11/042,816), filed Jan. 24, 2005 and
entitled "Chlorite in the Treatment of Neurodegenerative Disease";
and US Patent Pub. No. 2007/0145328, filed Dec. 21, 2006 and
entitled "Chlorite Formulations, and Methods of Preparation and Use
Thereof," all of which are incorporated herein by reference in
their entirety.
[0061] In one aspect, the present invention provides for the
treatment of a macrophage related disease using chlorite. The
chlorite ion is ClO.sub.2.sup.-. A chlorite (compound) is a
compound that contains this group, with chlorine in oxidation state
+3. Chlorites are also known as salts of chlorous acid. Chlorine
can assume oxidation states of -1, +1, +3, +5, or +7 within the
corresponding anions Cl.sup.-, ClO.sup.-, ClO.sub.2.sup.-,
ClO.sub.3.sup.-, or ClO.sub.4.sup.-, known commonly and
respectively as chloride, hypochlorite, chlorite, chlorate, and
perchlorate.
[0062] II. Tetrachlorodecaoxide (TCDO) and WF10
[0063] In some embodiments, the present invention provides methods
using one or more chlorite containing agents. The source of
chlorite ions for administration of chlorite according to the
present invention can be provided in a variety of forms. For
example, chlorite can be administered as a chlorite salt, for
example, alkali metal salt, e.g. sodium chlorite, potassium
chlorite, and the like, or a mixture of chlorite salts, where the
chlorite salts are preferably pharmaceutically acceptable. In
addition or alternatively, chlorite can be administered as a matrix
of chlorite ions, e.g., described in U.S. Pat. No. 4,507,285. In
one embodiment, the chlorite ions as provided in a composition
having the general formula
ClO.sub.2.times.nO.sub.2
wherein "n" can be a value of about 0.1-0.25. Such agents can have
an O.sub.2 band at 1562 cm.sup.-1 in the Raman spectrum and an O--O
interval of 123 pm. Production of such agents is known in the art,
see e.g., U.S. Pat. No. 4,507,285.
[0064] In one embodiment, the method of treatment involves
administration of an aqueous solution of a product known as
"tetrachlorodecaoxygen anion complex", commonly known as TCDO.
Production of TCDO is well known, see e.g., Example 1 of U.S. Pat.
No. 4,507,285. In some embodiments, the chlorite containing agents
that can be used in the methods of the present invention for
treating diabetes or related disorders include but are not limited
to chlorite salt, such as alkali metal salt, sodium chlorite,
potassium chlorite, and the like, a matrix of chlorite salts, a
matrix of chlorite ions, e.g., compositions having the general
formula ClO.sub.2.times.nO.sub.2, where "n" can be a value or about
0.1-0.25. One example is TCDO. One of the aqueous TCDO formulations
is WF10. WF10 is an aqueous formulation of the drug OXO-K993.
Oxoferin is a topical formulation of the same drug and is
registered and marketed as a wound healing agent in Europe and
Asia. WF10 is a sterile, pyrogen-free, aqueous 10% (w/v) solution
of OXO-K993 with no additional inactive ingredients and is intended
for intravenous infusion. TCDO is analytically characterized as a
solution containing 4.25% chlorite, 1.9% chloride, 1.5% chlorate,
0.7% sulfate, and sodium as the cation. The active principle is
defined by the chlorite ion content. In one embodiment, WF10
solution contains about 63 mmol/l of chlorite.
[0065] Tetrachlorodecaoxide (TCDO) is a chlorite-containing drug
used for the dressing of wounds, immunomodulation and as radiation
protective agent. Due to its oxidizing properties, TCDO can destroy
most pathogens although it is not regarded as antibiotic. But the
main reason for its use for dressing of wounds is not its
bactericidal activity. This drug is regarded as immunomodulating,
that is, it acts by stimulating the immune system of the body.
Tetrachlorodecaoxide combines with the heme part of hemoglobin,
myoglobin and peroxidase, forming a TCDO-hemo complex. This in turn
activates the macrophages and accelerates the process of
phagocytosis which engulfs most of the pathogens and cell debris
present on the surface of the wound, thus cleaning the wound
surface and helping in the regenerative process.
Tetrachlorodecaoxide is also mitogenic and chemotactic. The
mitogenic impulse gives rise to two factors, MDGF (Macrophage
derived growth factor) and WAF (Wound angiogenesis factor). The
MDGF deposits fibroblasts and synthesizes collagen fibers, which
fill the gap in the wounds, the WAF helps in the formation of new
capillaries which further enhances the healing process. The
chemotactic impulse acts on the myocyte (muscle cell) and causes it
to contract, thereby bringing the wound edges closer and reducing
the wound surface. Simultaneous influence of all these factors
accelerates the wound healing with minimal scarring.
[0066] WF10 is a 1:10 dilution of tetrachlorodecaoxide (TCDO)
formulated for intravenous injection. WF10 specifically targets
macrophages. WF10 potentially modulates disease-related
up-regulation of immune responses both in vitro and in vivo. Thus
immune response is influenced in a way that inappropriate
inflammatory reactions are downregulated (Arzneimittelforschung.
2001; 51(7):554-62. Schempp H, et al). WF10 is currently being
studied for treatment of late-stage HIV disease, as well as
recurrent prostate cancer, late post-radiation cystitis, autoimmune
disease and chronic active hepatitis C disease. WF10 is approved
for use in Thailand under the name IMMUNOKINE in patients with
post-radiation chronic inflammatory disease including cystitis,
proctitis and mucositis.
[0067] In vivo studies have investigated the effects of WF10 on
monocytes, macrophages and lymphocytes, on humoral and cellular
immunity, and on response to local or total body irradiation
(reviewed by McGrath M S et al. Current Opinion in Investigational
Drugs 2002 3(3)). WF10 increased the number of macrophages
infiltrating a skin blister in a human wound healing model (Hansel
M et al. Skin Pharmacol 1988 1:64). In rats, WF10 increased the
proportion of granulocytes, peripheral blood monocytes (PBMCs) and
large granular lymphocytes (LGLs), and stimulated erythropoiesis
after total body X-irradiation (Ivankovic S et al. OXO Study Report
1988 March; Ivankovic S et al. Radiat Res 1988 115: 115-123). In
mice, WF10 stimulated regeneration of hematopoietic stem cells
receiving sublethal doses of J-irradiation (Mason K A et al. Radiat
Res 1993 136: 229-235). In other studies, WF10 displayed direct
antitumor effects against radiation-induced, hemical-induced and
metastatic malignant and benign tumors (Kempf S R et al.
International Symposium on Tissue Repair 1990 Thailand; Milas L.
OXO Study Report 1991 September; Kempf S R et al. Radiat Res 1994
139: 226-231). WF10 altered proportions of T-helper and
suppressor/cytotoxic cells in spleen and thymus and increased both
the humoral and cellular immune responses (Gillissen G et al. OXO
Study Report 1993).
[0068] Without being bound by theory, it has been suggested that
WF10 causes marked inhibition of inducible genes related to T-cell
proliferation and cause reproducible upregulation of inflammatory
gene expression in macrophages in vitro, which is thought to
contribute to the higher rate of apoptosis in activated
macrophages. These data, coupled with an earlier report of WF10
inhibition of T-cell activation (McGrath M S et al. Transplant
Proc. 1998 30: 4200-4202), show that WF10 causes profound changes
in T-cell function through regulation of macrophage activation. The
WF 10 oxygen/chlorite matrix is stable until interaction with
heme-associated iron, whereupon it is converted to an active
chlorite molecule through a Michealis-Menten reaction and
intermediate production of a reactive compound I. Chlorite is the
active form of the drug thought to mediate the immunological
effects in macrophages.
[0069] A dose-ranging clinical study was conducted from 1993 to
1994 in 44 HIV-positive patients with <500 CD4+ T cells/mm
(Raffanti S P et al. Infection 1998 26: 201-206). The study
established the maximum tolerated dose as 0.5 ml/kg/day of WF10,
when administered in four 5-day cycles, with each cycle followed by
16 days of without treatment. No significant adverse events or
clinical laboratory toxicity were observed at this dosage. Plasma
CD8+ T-cell counts increased in a dose-dependent manner over four
cycles of WF10 administration. This study demonstrated that WF10 at
a dose of 0.5 ml/kg was associated with a sustained immunological
response, i.e., sustained elevation of CD8+ T cell numbers,
consistent with the proposed mechanism of action. Furthermore, a
single-center, phase I/II study, was conducted in 1997 to evaluate
safety and the effects of WF10 on the kinetics of red blood cell
(RBC) survival, selective immunological markers of HIV disease,
macrophage activation and viral kinetics (Herndier B et al.
Keystone Symposia on Molecular and Cellular Biology. 1998). Changes
in immunological parameters of cells from HIV+ patients in response
to WF10 treatment are summarized in Table 1 in McGrath M S et al.
Current Opinion in Investigational Drugs 2002 3(3), including an
increase in CD3+CD4+ cells, an increase in CD3+ CD8+ cells, an
increase in CD3+ CD4+ CD38- cells, an increase in CD3+ CD8+ CD38-
cells, an increase in CD3+ CD8+ CD28- cells, a decrease in CD3+
CD8+ CD28+ cells, a decrease in CD3+ CD4+ CD38+ cells, a decrease
in all CD14+ cells, and a decrease in CD20+ HLR-DR+ cells. The
results suggested that WF10 reduced antigen presentation while
concurrently inducing phagocytosis in macrophages with impaired
function. WF10 had no effect on HIV load over the course of the
trial. No significant differences were detected between the WF10
and placebo group in hematological and blood chemistry values,
including parameters specifically associated with hemolysis.
[0070] As appropriate, agents that provide a source of chlorite
ions can be administered in a free base or free acid form, i.e., as
the free compound and not as a salt. In some embodiments, the
chlorite formulation contains about 150 .mu.M chlorite.
[0071] Additionally, any pharmaceutically acceptable salt(s) of the
compound(s) can also be used. Pharmaceutically acceptable salts are
those salts which retain the biological activity of the free
compounds and which are not biologically or otherwise undesirable.
As appropriate, stereoisomers of the compounds disclosed can also
be used in the invention, including diastereomers and enantiomers,
as well as mixtures of stereoisomers, including but not limited to
racemic mixtures. Unless stereochemistry is explicitly indicated in
a structure, the structure is intended to embrace all possible
stereoisomers of the compound depicted.
[0072] In some embodiments, the oxidative compound is WF10. WF10 is
a chlorite-based compound. After interaction with heme proteins,
the chlorite matrix of WF10 acquires oxidizing and chlorinating
properties (Schempp H. et al. 1999). It has been suggested that
WF10 exerts potent immunomodulatory effects most likely through
generating physiologic oxidative compounds namely chloramines.
Chloramines have been reported to exert cell-protective and
anti-inflammatory activities (Choray M. et al. Amino Acids 23
(2002) 407-413).
[0073] Another stable oxidant that can be used in the methods of
the present invention is taurine-chloramine (TauCl), which is
produced by activated neutrophils in the course of inflammation and
was shown to possess cell-protective as well as anti-inflammatory
activities (Park E. et al. Clin. Immunol 102 (2002) 179-184). Its
precursor, taurine, represents the most abundant free amino acid
not incorporated into proteins in the cytosol of leukocytes.
Taurine acts as a scavenger for HOCl, a microbicidal agent produced
physiologically by the myeloperoxidase-hydrogen peroxide-halide
system of activated neutrophils and monocytes during the oxidative
burst (Cunningham C et al. Biochem J. 330 (1998) 939-945). The
reaction of taurine and HOCl forms taurine-chloramine, a long-lived
oxidant that is much less toxic than HOCl. TauCl itself represents
a potent biologic effector molecule, which contributes to
self-limitation of inflammatory processes (Marcinkiewicz J et al.
J. Leuko. Biol 58 (1995) 667-674). It has been demonstrated that
incubation of human peripheral mononuclear cells with WF10 leads to
a rapid and stable generation of chloramines (Giese T. et al.
Cellular Immunology, 229 (2004) 149-158) and TauCl represents the
most relevant functional product that is formed under the influence
of WF10.
[0074] In some embodiments, the present invention provides methods
of modulating macrophage activation or treating a macrophage
related disease comprising administering an effective amount of
taurine-chloramine into a subject. TauCl possesses the ability to
regulate the production of macrophage derived pro-inflammatory
mediators such as NO, IL-1, TNF-alpha, and PGE2. The mechanisms
involved include transcriptional as well as post-translational
regulation.
[0075] Pro-oxidative substances also have a direct effect on
transcriptional activities of the NFAT species of transcription
factors. The nuclear translocation of NFAT requires their
dephosphorylation by the calcium/calmodulin dependent
serine/threonine phosphatase calcineurin. The phosphatase activity
of calcineurin is redox sensitive. WF10 is able to inhibit antigen
receptor driven lymphocyte proliferation. Expression of NFAT
regulated genes is strongly suppressed by WF10, and the nuclear
translocation of NFATc is inhibited. The WF10 associated inhibition
of NFAT regulated genes in activated T cells, in concert with the
induction of several monocyte associated pro-inflammatory genes,
suggest activation of the innate myeloid functions concomitant with
the inactivation of adaptive proliferative lymphocyte response.
This approach represents a novel method of targeting
redox-regulation for the treatment of inflammatory disorders. In
some embodiments, the macrophage related diseases that can be
treated using the methods of the present invention are inflammatory
diseases.
[0076] III. Chlorite Purity and pH
[0077] Methods of formulating chlorite have been described in
described in US Patent Pub. No. 20070145328, filed Dec. 21, 2006
and entitled "Chlorite Formulations, and Methods of Preparation and
Use Thereof," which is incorporated herein by reference in its
entirety. Such formulations are suitable for various modes of
administration, including but not limited to non-topical,
parenteral, systemic, or intravenous administration.
[0078] In some embodiments, the present invention makes use of
chlorite formulated in aqueous solution in which the chlorite is
97-99% pure. As used herein, the "purity" of chlorite in a sample
is calculated as the percent weight of chlorite salt to the total
weight of the sample. In determining the purity of chlorite in a
solution, the weight of the solvent (e.g., water in an aqueous
solution) is not included. Purity may be evaluated using ion
chromatography and an ion detector, by calibrated integration of
the respective peaks; for example, chlorite, chloride, chlorate,
phosphate and sulfate in the compound or formulation. For example,
chlorite is commercially available as sodium chlorite, technical
grade, at a purity of 80% (catalog No. 244155 Sigma-Aldrich).
[0079] Alternatively, cyrstalline sodium chlorite is provided in a
purity greater than 95%, greater than 96%, greater than 97%,
greater than 98%, greater than 99%, or greater than 99.9%. Solid
pharmaceutical formulations comprising crystalline sodium chlorite
in a purity greater than 95%, greater than 96%, greater than 97%,
greater than 98%, greater than 99%, or greater than 99.9% in
addition to one or more pharmaceutical excipients are also
encompassed.
[0080] In some embodiments, the chlorite formulations for use with
the present invention comprise low amounts of chlorate, sulfate or
chloride. As used herein, a formulation is "substantially free" of
a molecule if the molecule comprises no more than 1 part in 1000
per weight of non-solvent molecules in the formulation. In certain
embodiments, the weight ratio of chlorite to chlorate is greater
than 100:1.5, greater than 100:0.5, greater than 100:1, or greater
than 100:0.1. In one embodiment, the composition is substantially
free of chlorate. In another embodiment, the weight ratio of
chlorite to chloride is greater than 100:45.5 or greater than
100:8.5. In one embodiment the composition is substantially free of
chloride. In a further embodiment, the weight ratio of chlorite to
sulfate is greater than 100:16.4 or greater than 100:1.6. In one
embodiment the composition is substantially free of sulfate.
[0081] The pH of a chlorite formulation for use with the present
invention can be adjusted to between about 7 and about 11.5. In
some embodiments, the pH of a chlorite formulation is lowered to
between about 7 and about 11.5 using a pH adjusting compound that
does not expose the formulation to high local acidity. In some
embodiments, the pH adjusting compound is any one or more of
monosodium phosphate, disodium phosphate, or acetic acid.
[0082] Also described herein are methods of preparing chlorite
formulations and pharmaceutical formulations, including but not
limited to the chlorite formulations specifically described herein.
Also described herein are kits and methods of administration of the
formulations and pharmaceutical formulations described herein.
Various exemplary aspects and variations of the invention are
described in the "Brief Summary of the Invention," as well as
elsewhere herein, including but not limited to the Examples. It is
also understood that the invention includes embodiments comprising,
consisting essentially of, and/or consisting of one or more
elements as described herein.
[0083] In some embodiments, the invention makes use of aqueous
formulations comprising chlorite. In some embodiments, the chlorite
formulation comprises an aqueous solvent, and optionally one or
more other solvents for chlorite. In some embodiments, the
formulations comprise chlorite and an aqueous solvent for chlorite,
and have a pH of about 7 to about 11.5.
[0084] Solvents or combinations of solvents for use in the
formulations described herein can be determined by a variety of
methods known in the art. One nonlimiting example includes (1)
theoretically estimating solvent solubility parameter value(s) and
choosing the one(s) that match with chlorite, using standard
equations in the field; and (2) experimentally determining the
saturation solubility of chlorite in the solvent(s), and (3)
choosing one or more that exhibits the desired solubility, and (4)
selecting a solvent or solvents that do not diminish the activity
of chlorite, or that do not or only minimally react with chlorite.
In some embodiments, the liquid formulations described herein
comprise a plurality of solvents.
[0085] In some embodiments, the chlorite formulations comprise an
aqueous solvent. In some variations, water is the principal solvent
in the aqueous formulations. In some variations, water is at least
about 50% by volume of the solvent component of an aqueous
formulation. In some variations, water is at least about 50% by
volume of the aqueous formulation. In some variations, water is any
of between about 50 to about 60, between about 60 to about 70,
between about 70 to about 80, between about 80 to about 90, between
about 90 to about 99, at least about 50, at least about 60, at
least about 70, at least about 80, at least about 90, or at least
about 95, about 50, about 60, about 70, about 80, about 90, or
about 95 percent by volume of the solvent component. In some
variations, water is any of between about 50 to about 60, between
about 60 to about 70, between about 70 to about 80, between about
80 to about 90, between about 90 to about 99, at least about 50, at
least about 60, at least about 70, at least about 80, at least
about 90, or at least about 95, percent by volume of the aqueous
formulation. In some variations, water is at least about 95% by
volume of the aqueous formulation. In some variations, water is
between about 80 to about 90% by volume of the aqueous formulation.
In some variations, water is between about 90 to about 99% by
volume of the aqueous formulation.
[0086] The formulations may have differing concentration of
chlorite. In some embodiments, the concentration of chlorite in the
formulation is high, and then is diluted to a less concentrated
form prior to administration. In some embodiments, a formulation
described herein is diluted any of about 2.5.times., about
5.times., about 7.5.times., about 10.times., about 20.times., about
25.times., about 50.times., about 100.times., about 200.times.,
about 250.times., about 300.times., about 500.times., or about
1000.times.. In some embodiments, a formulation described herein is
diluted about 2.5.times., about 5.times., about 10.times., about
20.times., about 25.times., about 50.times., about 100.times.,
about 200.times., about 250.times., about 300.times., about
500.times., about 1000.times.; between about 2.times. and about
10.times., between about 10.times. and about 50.times., between
about 50.times. and about 100.times., between about 100.times. and
about 500.times., or between about 500.times. and about
1000.times.. In some embodiments, a formulation as described herein
is diluted between about 2.times. and about 10.times.. In some
embodiments, a formulation as described herein is diluted between
about 10.times. and about 50.times.. In some embodiments, a
formulation as described herein is diluted about 7.5.times.. In
some embodiments, a formulation as described herein is diluted
about 25.times.. In some embodiments, a formulation as described
herein is diluted about 200.times.. In some embodiments, the
concentration of chlorite in the formulations described herein is
between about 1 .mu.M and about 1.5 M. In another embodiments, the
concentration of chlorite in the formulations described herein is
between any of about 1 M and about 1.5 M; between about 1 .mu.M and
about 100 mM; between about between about 10 .mu.M and about 100
mM; between about 0.1 mM and about 10 mM; between about 0.1 mM and
about 500 mM; between about 0.1 mM and about 200 mM; between about
1 mM and about 100 mM; between about 0.1 mM and about 5 mM; between
about 50 mM and about 100 mM; between about 55 mM and about 70 mM;
between about 60 mM and about 65 mM; between about 100 mM and about
500 mM; between about 200 mM and about 400 mM; between about 300 mM
and about 700 mM; about 1 mM; about 1.5 mM; about 2 mM; about 2.5
mM; about 3 mM; about 3.5 mM; about 4 mM; about 5 mM; about 10 mM;
about 20 mM; about 30 mM; about 40 mM; about 50 mM; about 60 mM;
about 62 mM; about 65 mM; about 70 mM; about 80 mM; about 90 mM;
about 100 mM; at least about 0.1 mM; at least about 1 mM; at least
about 2 mM; at least about 5 mM; at least about 10 mM; at least
about 20 mM; at least about 30 mM; at least about 40 mM; at least
about 50 mM; at least about 60 mM; at least about 70 mM; at least
about 80 mM; at least about 90 mM; or at least about 100 mM. In
some embodiments, the concentration of chlorate in the formulations
described herein is between about 50 mM and about 100 mM. In some
embodiments, the concentration of chlorate in the formulations
described herein is between about 55 mM and about 75 mM. In some
embodiments, the concentration of chlorate in the formulations
described herein is between about 0.1 mM and about 10 mM. In some
embodiments, the concentration of chlorate in the formulations
described herein is between about 1 mM and about 5 mM.
[0087] In some embodiments, the chlorite formulation has a pH no
greater than about 12.0. In some embodiments, the pH of the
formulation is any of no greater than about 11.5, about 11.0, about
10.5, about 10.0, about 9.5, about 9.0, about 8.5, about 8.0, about
7.5, about 7.0, about 6.5, or about 6.0. In some embodiments, the
pH of the formulation is no greater than about 11.5. In some
embodiments, the pH of the formulation is no greater than about
10.5. In some embodiments, the pH of the formulation is no greater
than about 8.5. In some embodiments, the pH of the formulation is
no greater than about 7.5. In some embodiments, the pH of the
formulation is between any one or more of about 7 and about 12;
between about 7 and about 11.5; between about 7 and about 10.5;
between about 7 and about 10; between about 7 and about 9.5;
between about 7 and about 9.0; between about 7 and about 8.5;
between about 7 and about 8.0; between about 7 and about 7.5;
between about 7.5 and about 8; between about 7.5 and about 8.5;
between about 7 and about 8; between about 8 and about 9; between
about 7.0 and about 8.5; between about 8 and about 8.5; between
about 8.5 and about 9; between about 7.1 and about 7.7; between
about 7.2 and about 7.6; between about 7.3 and about 7.4; about
7.0; about 7.1; about 7.2; about 7.3; about 7.4; about 7.5; about
7.6; about 7.7; about 7.8; about 7.9; about 8.0; about 8.1; about
8.2; about 8.3; about 8.4; about 8.5; about 8.6; about 8.7; about
8.8; or about 8.9. In some embodiments, the chlorite formulation
has a pH of about 7.0 to about 9.0. In some embodiments, the
chlorite formulation has a pH of about 7.0 to about 8.5. In some
embodiments, the chlorite formulation has a pH of about 6.0 to
about 8.5. In some embodiments, the chlorite formulation has a pH
of about 7.0 to about 8.0. In some embodiments, the chlorite
formulation has a pH of about 7.4.
[0088] In some embodiments, the chlorite formulations have a pH as
described above, and are formulated for any one or more of
parenteral, systemic, or intravenous administration.
[0089] In some embodiments, the chlorite formulations have a pH as
described above, and have a percentage chlorite purity as described
herein.
[0090] In some embodiments, the formulations described herein have
a pH as described above, and have a concentration of chlorite as
described herein. In some embodiments, the aqueous formulations
described herein have a pH between about 7 and about 11.5, or
between about 7.0 and about 10, or between about 7.0 and about 9.0,
or between about 7.0 and about 8.5, or between about 7.1 and about
7.7, and have a concentration of chlorite between about 1 and about
100 mM. In some embodiments, the aqueous formulations described
herein have a pH between about 7 and about 11.5, or between about
7.0 and about 10, or between about 7.0 and about 9.0, or between
about 7.0 and about 8.5, or between about 7.1 and about 7.7, and
have a concentration of chlorite between about 1 and about 5 mM. In
some embodiments, the aqueous formulations described herein have a
pH between about 7 and about 11.5, or between about 7.0 and about
10, or between about 7.0 and about 9.0, or between about 7.0 and
about 8.5, or between about 7.1 and about 7.7, and have a
concentration of chlorite between about 50 and about 80 mM.
[0091] In some embodiments, the aqueous formulations described
herein have a pH between about 7 and about 11.5, or between about
7.0 and about 10, or between about 7.0 and about 9.0, or between
about 7.0 and about 8.5, or between about 7.1 and about 7.7,
wherein the pH was adjusted with a pH adjusting agent that is any
one or more of a phosphate, or acetic acid.
[0092] In some embodiments, the formulations described herein are
stable with respect to one or more of pH or chlorite degradation
over a period of any of at least about 1 day, at least about 2
days, at least about 3 days, at least about 4 days, at least about
5 days, at least about 6 days, at least about 1 week, at least
about 2 weeks, at least about 3 weeks, at least about 4 weeks, at
least about 5 weeks, at least about 6 weeks, at least about 7
weeks, at least about 8 weeks, at least about 1 month, at least
about 2 months, at least about 3 months, at least about 4 months,
at least about 5 months, or at least about 6 months. In some
embodiments, the formulations described herein are stable with
respect to one or more of pH or chlorite degradation over a period
of any of at least about 1 week. In some embodiments, the
formulations are stable with respect to one or more of pH or
chlorite degradation over a period of any of at least about 1
month. In some embodiments, the formulations described herein are
stable with respect to one or more of pH or chlorite degradation at
one or more of room temperature, refrigerated conditions, or
approximately 4.degree. C. In some embodiments, the formulations
described herein are stable with respect to one or more of pH or
chlorite degradation under conditions of diminished light or
storage in a container that limits the amount of light to which the
formulation is subjected. In some embodiments, the formulations
described herein are stable with respect to one or more of pH or
chlorite degradation when stored in the dark. Examples of stable
pH, as used herein, means that the pH of the formulation changes by
less than any of about 0.1, about 0.2, about 0.3, about 0.4, about
0.5, about 0.6, about 0.7, about 0.8, about 0.9, or about 1
relative to the pH of the formulation as initially prepared. In
some embodiments, the pH of the formulation changes by less than
about 0.2 relative to the pH of the formulation as initially
prepared. The pH may be measured using, for example, a pH meter.
Examples of stable chlorite formulations include those in which
less than any of about 0.1%, less than about 0.2%, less than about
0.3%, less than about 0.4%, less than about 0.5%, less than about
0.6%, less than about 0.7%, less than about 0.8%, less than about
0.9%, less than about 1%, less than about 2%, less than about 3%,
less than about 4%, less than about 5%, less than about 6%, less
than about 7%, less than about 8%, less than about 9%, or less than
about 10% of the chlorite degrades into a non-chlorite ion relative
to the amount of chlorite present in the formulation as initially
prepared. In some embodiments, less than about 2% of the chlorite
degrades into a non-chlorite compound relative to the amount of
chlorite present in the formulation as initially prepared. In some
embodiments, less than about 0.5% of the chlorite degrades into a
non-chlorite compound relative to the amount of chlorite present in
the formulation as initially prepared. The presence of non-chlorite
elements may be measured, for example, using gas chromatography
(GC), mass spectrometry, or other methods known by those of skill
in the art.
[0093] In some embodiments, the chlorite formulations described
herein comprise no greater than about 5% by weight of deleterious
non-chlorite elements of other commercially available formulations.
In some embodiments, the chlorite formulations described herein
comprise any of no greater than about 4%, about 3%, about 2%, about
1%, about 0.5%, about 0.3%, about 0.25%,about 0.2%, about 0.1%,
about 0.05%, or about 0.02%, by weight of deleterious non-chlorite
elements of other commercially available formulations. In some
embodiments, the chlorite formulations described herein comprise
any of no greater than about 4% by weight of deleterious
non-chlorite elements of other commercially available formulations.
In some embodiments, the chlorite formulations described herein
comprise any of no greater than about 2% by weight of deleterious
non-chlorite elements of other commercially available formulations.
In some embodiments, the chlorite formulations described herein
comprise any of no greater than about 0.5% by weight of deleterious
non-chlorite elements of other commercially available formulations.
In some embodiments, the chlorite formulations described herein
comprise any of no greater than about 0.05% by weight of
deleterious non-chlorite elements of other commercially available
formulations. In some embodiments, the chlorite formulations
described herein are substantially free of the deleterious
non-chlorite elements of other commercially available formulations.
Nonlimiting examples of methods of detection of non-chlorite
components include HPLC; SPCS, for example using a Novosep A2
column with 3.6 mM Sodium Carbonate as a mobile phase, 5.mu.,
250.times.4.0 mm, flow rate 0.8 mL/min; DS-Plus Suppressor, for
example using a Novosep A2 column with 3.6 mM Sodium Carbonate as a
mobile phase, 5.mu., 250.times.4.0 mm, flow rate 0 8 mL/min; an
Allsep A-2 Anion column using 2.1mM NaHCO.sub.3/1.6 mM
Na.sub.2CO.sub.3 as a mobile phase, 100.times.4.6 mm, flow rate 2.0
mL/min; an anion HC column using 2.8 mM NaHCO.sub.3:2.2 mM
Na.sub.2CO.sub.3 in 10% Methanol as a mobile phase, 150.times.4.6
mm, flow rate 1.4 mL/min; or an Allsep A-2 Anion column using 2.1
mM NaHCO.sub.3/1.6 mM Na.sub.2CO.sub.3 as a mobile phase, 5.mu.,
100.times.4.6 mm, flow rate 1.0 mL/min. See, for example, the
Alltech Associates, Inc. Grace Davison line of products and product
information for details. In some embodiments, formulations
described herein comprise no greater than about 10%, about 20%,
about 30%, about 40%, about 50%, about 60%, about 70%, about 75%,
about 80%, about 85%, about 90%, or about 95% of the amount of a
member of the group consisting of, or alternatively any one or more
of, chlorate ions and sulfate ions present in an equal
weight/volume percent of chlorite formulated as WF10 or a dilution
thereof. That is, in some embodiments, when a non-WF10 formulation
as described herein comprises a certain percent w/v of chlorite,
such formulation has no greater than about the stated percentage of
the amount of one or more of the specified non-chlorite components
in WF10 or a dilution thereof, wherein the WF10 or dilution thereof
comprises the same percent w/v of chlorite as is found in the
non-WF10 formulation with which it is being compared. In some
embodiments, the formulations described herein comprise no greater
than about 75% of the amount of a member of the group consisting
of, or alternatively any one or more of, chlorate ions and sulfate
ions present in an equal weight/volume percent of chlorite
formulated as WF10. In some embodiments, the formulations described
herein comprise no greater than about 85% of the amount of a member
of the group consisting of, or alternatively any one or more of,
chlorate ions and sulfate ions present in an equal weight/volume
percent of chlorite formulated as WF10. In some embodiments, the
formulations described herein comprise no greater than about 50% of
the amount of a member of the group consisting of, or alternatively
any one or more of, chlorate ions and sulfate ions present in an
equal weight/volume percent of chlorite formulated as WF10.
[0094] It can be understood from the product insert of WF10 that
WF10 reportedly includes a ratio of chlorite to chlorate of
100:35.7 (4.25% to 1.5%), a ratio of chlorite to chloride of
100:45.5 (4.25% to 1.9%) and a chlorite to sulfate ratio of
100:16.4 (4.25% to 0.7%).
[0095] Examples of deleterious non-chlorite components include
non-chlorite components that cause an adverse reaction when
administered to physiological systems. In some variations, a
deleterious non chlorite component is associated with one or more
indicia of toxicity in one or more of in vitro or in vivo assays
known in the art, or are associated with one or indicia of toxicity
when administered to a physiological system, including but not
limited to a subject, including but not limited to a human subject.
Deleterious non chlorite components include but are not limited to
sulfate, chlorine dioxide, chlorate, and borate. In some
embodiments, the chlorite formulations described herein are
substantially free of the deleterious non-chlorite elements of
WF10. In some variations, the chlorite formulations described
herein are substantially free of sulfate and chlorate ions.
[0096] In some embodiments, the chlorite formulations described
herein contain less than about 1.9% of chloride ions. In some
embodiments, the chlorite formulation contains any of less than
about 1.9%, less than about 1.8%; less than about 1.5%; less than
about 1.0%; less than about 0.5%; less than about 0.3%; less than
about 0.1%; less than about 0.05%; less than about 0.01%; less than
about 0.001%; between about 0.001 to about 0.1%; between about 0.1
to about 0.5%; between about 0.5 to about 1.0%; between about 1.0
to about 1.5%; or between about 1.5 to about 1.8%by weight of
chloride ions. In some embodiments, the chlorite formulation
contains less than about 0.5% by weight of chloride ions. In some
embodiments, the chlorite formulation contains less than about
0.24% by weight of chloride ions. In some embodiments, the chlorite
formulation contains less than about 0.2% by weight of chloride
ions. In some embodiments, the chlorite formulation contains less
than about 0.1% by weight of chloride ions. In some embodiments,
the chlorite formulation is substantially free of chloride ions. In
some embodiments, the level of chloride ions is below the level of
detection using HPLC.
[0097] In some embodiments, the chlorite formulation contains less
than about 1.5% of chlorate ions. In some embodiments, the chlorite
formulation contains any of less than about 1.4%, less than about
1.3%; less than about 1.0%; less than about 0.5%; less than about
0.3%; less than about 0.1%; less than about 0.01%; less than about
0.001%; between about 0.001 to about 0.1%; between about 0.001 to
about 0.01%; between about 0.01 to about 0.1%; between about 0.1 to
about 0.5%; between about 0.5 to about 1.0%; or between about 1.0
to about 1.4% of chlorate ions. In some embodiments, the chlorite
formulation is substantially free of chlorate ions. In some
embodiments, the chlorite formulation contains less than about 0.5%
by weight of chlorate ions. In some variations, the chlorite
formulation is substantially free of chlorate ions. In some
embodiments, the chlorite formulation contains less than about
0.19% by weight of chlorate ions. In some embodiments, the chlorite
formulation contains less than about 0.1% by weight of chlorate
ions. In some embodiments, the level of chlorate ions is below the
level of detection using HPLC.
[0098] In some embodiments, the chlorite formulation contains less
than about 0.7% of sulfate ions. In some embodiments, the chlorite
formulation contains any of less than about 0.65%; less than about
0.6%; less than about 0.5%; less than about 0.4%; less than about
0.3%; less than about 0.2%; less than about 0.1%; less than about
0.08%; less than about 0.07%; less than about 0.06%; less than
about 0.05%; less than about 0.005%; less than about 0.0005%;
between about 0.001 to about 0.1%; between about 0.01 to about
0.1%; between about 0.01 to about 0.5%; between about 0.06 to about
0.08%; or between about 0.5 to about 0.65% of sulfate ions. In some
embodiments, the chlorite formulation contains between about 0.5 to
about 0.65% of sulfate ions. In some embodiments, the chlorite
formulation is substantially free of sulfate ions. In some
embodiments, the chlorite formulation contains less than about 0.5%
by weight of sulfate ions. In some embodiments, the chlorite
formulation is substantially free of sulfate ions. In some
embodiments, the chlorite formulation contains less than about
0.08% by weight of sulfate ions. In some embodiments, the level of
sulfate ions is below the level of detection using HPLC.
[0099] In some embodiments, the chlorite formulations described
herein comprise phosphate ions. In some embodiments, the chlorite
formulations described herein comprise sodium ions. In some
embodiments, a chlorite formulation comprises chlorite, an aqueous
solvent, sodium, and phosphate ions. In some variations, the
aqueous solvent consists essentially of water. In some embodiments,
a chlorite formulation consists essentially of chlorite, water,
sodium, and phosphate, and is substantially free of chlorate. In
some embodiments, a chlorite formulation consists essentially of
chlorite, water, sodium, and phosphate, and is substantially free
of chlorate, and further comprises a pharmaceutically acceptable
diluent. In some embodiments, sodium and phosphate are provided in
whole or in part as monosodium phosphate or disodium phosphate. In
some embodiments, the pharmaceutically acceptable diluent is a
saline solution.
[0100] In some embodiments, the chlorite formulations described
herein comprise no greater than about 10% by weight of by products
or impurities present in commercially available technical grade
chlorite. Nonlimiting examples of by-products or impurities present
in commercially available technical grade chlorite include
chlorate, sulfate, chlorine dioxide, chloride, sodium bicarbonate,
and sodium carbonate. In some embodiments, the chlorite
formulations described herein comprise no greater than about any of
15%, about 12%, about 10%, about 9%, about 8%, about 7%, about 6%,
about 5%, about 4%, about 3%, about 2%, about 1%, about 0.5%, about
0.3%, about 0.1%, between about 0.1 to about 5%; between about 5 to
about 10%; or between about 10 to about 15% by weight of one or
more degradation products or impurities present in commercially
available technical grade chlorite, including but not limited to
one or more of chlorate or sulfate. In some embodiments, the
chlorite formulations described herein comprise no greater than
about 0.5% by weight of degradation products or impurities present
in commercially available technical grade chlorite, including but
not limited to one or more of chlorate or sulfate. In some
embodiments, the chlorite formulations described herein comprise no
greater than about 5% by weight of degradation products or
impurities present in commercially available technical grade
chlorite, including but not limited to one or more of chlorate or
sulfate. In some embodiments, the chlorite formulations described
herein are substantially free of the degradation products or
impurities present in commercially available technical grade
chlorite, including but not limited to chlorate or sulfate.
[0101] In some embodiments, the formulations described herein are
less toxic to a subject than previously reported chlorite
formulations at the same concentration of chlorite, when
administered by at least one of the routes of administration
described herein, including but not limited to by non-topical,
systemic, parenteral, or intravenous administration. In some
embodiments, the toxicity of a chlorite formulation is analyzed for
toxicity using an in vivo or in vitro toxicity assay, including
well-known toxicity assays. In some embodiments, the chlorite
formulation is analyzed for toxicity using a non-specific in vitro
toxicity assay.
[0102] In another variation, toxicity is measured according to
various response indicia of toxicity in a subject after
administration of the chlorite formulations described herein, as
compared to administration of other commercially available chlorite
formulations. In some variations, toxicity is measured relative to
systemic administration of chlorite formulated as WF10. In another
variation, toxicity is measured relative to intravenous
administration of chlorite formulated as WF10 to a subject. In some
variations, toxicity is measured after administration to a
mammalian subject, including but not limited to a human subject. In
some variations, toxicity is measured as one or more of irritation
to the surface to which the chlorite formulation is exposed,
including but not limited to the gastrointestinal tract, nausea,
vomiting, diarrhea, abdominal pain, hemolysis, methemoglobinemia,
cyanosis, anuria, coma, convulsions, liver damage, kidney damage,
loss of appetite, or weight loss. In some variations, toxicity is
measured as one or more of asthenia, injection site pain, headache,
rhinitis, or diarrhea. See, e.g., McGrath M S, "Development of
WF10, A Novel Macrophage-Regulating Agent," Curr Opin Investig
Drugs, 3(3):365-73 (March 2002), which is incorporated by reference
in its entirety. In another variation, toxicity is measured as
anemia. See, e.g., Kempf et al., "Comparative Study on the Effects
of Chlorite Oxygen Reaction Product TCDO (Tetrachlorodecaoxygen)
and Sodium Chlorite Solution (NaClO2) With Equimolar Chlorite
Content on Bone Marrow and Peripheral Blood of BDIX Rats," Drugs
Under Experimental and Clinical Research, 19(4):165-1 (1993). In
some variations, toxicity is measured as asthenia. In some
variations, toxicity is measured as injection site reaction. In
some variations, toxicity is measured as injection site pain.
[0103] IV. Methods of Purifying Chlorite
[0104] Various methods can be used for purifying chlorite, e.g., to
produce the formulations or pharmaceutical formulations described
herein. However, the formulations and pharmaceutical formulations
described herein may also be produced by other methods, and the
formulations and pharmaceutical formulations described herein are
not limited to those produced by the methods described herein.
[0105] In some embodiments, the purification is by subjecting a
mixture comprising chlorite to conditions in which chlorite is in
solution but one or more impurities are insoluble. The chlorite is
separated from the insoluble impurities. In some embodiments, the
chlorite is further purified by crystallization of the chlorite
from the mixture, and separation of the chlorite from the remaining
mixture. In some embodiments, the chlorite is purified from a
mixture comprising sodium chlorite.
[0106] Generally, the chlorite ions may be from any source
containing chlorite. For example, chlorite may be a chlorite salt,
for example alkali metal salts, sodium chlorite, potassium
chlorite, and the like, or a mixture of chlorite salts.
Alternatively, the source of chlorite may be from a formulation
comprising chlorite. In some embodiments, chlorite is purified from
a formulation comprising TCDO or WF10. In some embodiments,
chlorite is from a solution comprising sodium chlorite.
[0107] In some embodiments, impure chlorite, including but not
limited to impure sodium chlorite, is dissolved in a solvent or a
solvent system. In some variations, any solvent in which chlorite
dissolves is used. In some embodiments, any solvent in which
chlorite dissolves and with which chlorite does not react is used.
In some embodiments, the solvent is distilled water. In some
embodiments, the solvent is a non-organic solvent.
[0108] In some embodiments, impure sodium chlorite is between about
0.1% to about 99% per weight of the starting material. As
non-limiting example of the purity of the chlorite starting
material, the chlorite is between about any of 0.1% and about 5%;
between about 1% and about 5%; between about 4% and about 10%;
between about 1% and about 15%; between about 15% and about 25%;
between about 5% and about 25%; between about 25% and about 50%;
between about 50% and about 75%; between about 75% and about 85%;
between about 85% to about 95%; between about 60% and about 90%;
between about 95% and about 99% per weight of the starting
material; at least about 50, at least about 60, at least about 70,
at least about 80, at least about 90, or at least about 95% pure.
If the impure chlorite is in a solvent, the percent purity is
relative to the non-solvent components. In some embodiments, the
chlorite is between about 75% and about 85% pure. In some
embodiments, the chlorite is between about 85% and about 95% pure.
In some embodiments, the chlorite is at least about 85% pure.
[0109] In some embodiments, small amounts of hydrogen peroxide are
added to the dissolved chlorite. While not bound by theory, the
addition of small amounts of hydrogen peroxide may reduce sodium
chlorate to sodium chlorite. If desired, unreacted hydrogen
peroxide may be subsequently removed. In some embodiments, hydrogen
peroxide is added after the initial dissolution of chlorite step
and subsequently removed by filtration, for example, by centrifugal
filtration.
[0110] In some embodiments, an impure solution of chlorite is
subjected to conditions wherein the chlorite remains soluble, but
one or more of the impurities is no longer soluble. One such method
is described in Russian Patent No. SU327132, which is hereby
incorporated by reference in its entirety. In some variations, the
impure solution is concentrated at an elevated temperature until
one or more impurities precipitate. It is envisioned that the
precipitated impurities can include but are not limited to
chlorate, chloride, and sulfate. In some embodiments, the impure
solution is concentrated at a temperature that is any of between
about 60 to about 100.degree. C.; between about 65 to about
75.degree. C.; between about 60 to about 80.degree. C.; between
about 60 to about 100.degree. C.; between about 70 to about
90.degree. C.; at about 60.degree. C.; at about 65.degree. C.; at
about 70.degree. C.; at about 75.degree. C.; or at about 80.degree.
C. In some variations, the impure solution is concentrated at a
temperature that is between about 65 to about 75.degree. C.
[0111] In some embodiments, the impure solution is concentrated
using the methods described herein and under reduced pressure. A
skilled artisan is familiar with a range of suitable techniques for
providing reduced pressure including but not limited to the
application of a vacuum during concentration.
[0112] The degree to which the chlorite solution is concentrated
may be varied. In some variations, the solid to liquid phase volume
ratio in the suspension is no greater than about 1 part to about 12
parts. By way of one non-limiting example, at least 50% water
removal from a starting solution of chlorite has been demonstrated
to result in the elimination of a significant amount of impurities,
e.g., chloride and/or chlorate, while maintaining chlorite in
solution. One or more impurities are thereafter separated from the
chlorite. In some variations, the impurities are removed while the
mixture is still subject to conditions wherein the chlorite remains
soluble, but one or more of the impurities are no longer soluble.
One method of removing the impurities is by filtration. If
filtration is used, the temperature upon filtration may be, e.g.,
at an elevated temperature that is similar to the temperature at
which the concentration was performed. In some variations
filtration is used, and the filtration occurs shortly after
concentration. The concentrated chlorite solution includes greater
than 80% pure chlorite by weight. The purity of chlorite in the
concentrated solution can be greater than 85% pure or greater than
90% pure by weight.
[0113] The resulting chlorite solution may optionally be again
subjected to conditions wherein the chlorite remains soluble, but
one or more of the impurities is no longer soluble. The conditions
may be optimized to reduce the same or a different impurity as was
reduced in the first purification.
[0114] In some embodiments, the chlorite is subjected to conditions
wherein the chlorite is not soluble, but the impurities are
soluble. In some embodiments, chlorite is purified by inducing
chlorite to crystallize from a solution. In some embodiments, the
chlorite is induced to crystallize from a concentrated filtrate
prepared by the methods described herein. As one non-limiting
example, chlorite may be induced to crystallize by cooling the
chlorite solution. In some variations, chlorite is induced to
crystallize by exposure of a chlorite solution to a temperature
that is any of no greater than about 10.degree. C., no greater than
about 0.degree. C., no greater than about -10.degree. C.; no
greater than about -20.degree. C.; no greater than about
-30.degree. C.; no greater than about -40.degree. C.; between about
-15.degree. C. and about -35.degree. C., between about -20.degree.
C. and about -30.degree. C.; about 10.degree. C.; about 0.degree.
C.; about -10.degree. C.; about -15.degree. C.; about -20.degree.
C.; about -25.degree. C.; about -30.degree. C.; about -35.degree.
C.; or about -40.degree. C. In some embodiments, the chlorite is
induced to crystallize by exposure of a chlorite solution to a
temperature no greater than about -20.degree. C. In some
embodiments, the chlorite is induced to crystallize by exposure of
a chlorite solution to a temperature between about -20.degree. C.
and about -30.degree. C.
[0115] In general, the chlorite may be cooled at different rates,
such as a stepwise placement into increasingly cool environments,
or the chlorite formulation may be placed in a single cooling
environment. The chlorite formulation may be cooled over a period
of, for example, 12 to 24 hours. Longer or shorter periods may also
be utilized. In some embodiments, the chlorite formulation is
cooled over a period of between about 12 and about 14, about 14 and
about 16, about 16 and about 18, about 18 and about 20, about 20
and about 22, or about 22 to about 24 hours.
[0116] In some embodiments, chlorite is harvested from a mixture
comprising chlorite solids. The solids may be harvested by various
methods known by those of skill in the art, including but not
limited to by filtration. In some variations, chlorite solids are
harvested from a mixture comprising chlorite that has been cooled
to crystallize the chlorite solids.
[0117] In some embodiments, a mixture comprising chlorite solids is
filtered as a frozen mixture comprising chlorite melts. When
filtration is specified, those of skill in the art can determine an
appropriate method of filtration. In some embodiments, suction
filtration is used to separate chlorite solids from a mixture. The
chlorite solids can be in the form of a slurry or slush. In another
embodiment, centrifugal filtration is used to separate chlorite
solids from a mixture. During centrifugal centrifugation, water
(ice) remaining in the mixture containing chlorite solids melts and
can be eliminated while the chlorite solids remain associated with
a filter. In some embodiments, centrifugal filtration is performed
using a 50 micron filter at 1200 rpm. In some embodiments, the
filter is about 50 to about 250 microns. In some embodiments, the
rpm are about 1000 to about 3600. In some embodiments, the force of
gravity is used to separate the solids from the rest of the
mixture. The resulting chlorite solids can be in a hydrated
form.
[0118] The chlorite may optionally be recrystallized by the above
or a different method. In some embodiments, the chlorite is
recrystallized to give chlorite of increased purity relative to the
first recrystallization.
[0119] The present invention can use mixtures comprising chlorite,
wherein the chlorite is any of greater than about 70%, greater than
about 80%, greater than about 85%, greater than about 90%, greater
than about 95%, greater than about 99%; between about 70-80%;
between about 80-90%; between about 90-99%; about 70%; about 75%;
about 80%; about 85%; about 90%; about 95%; or about 99% pure. If a
solvent is present, including but not limited to water, the
percentage purity is relative to the non-solvent components. In
some embodiments, the chlorite is between about 70-80% pure. In
some embodiments, the chlorite is between about 80-90% pure.
[0120] Purified chlorite may be dissolved in an aqueous solvent to
give a chlorite solution of the desired concentration or molarity.
As one example, the purified chlorite may be dissolved in distilled
water or a saline solution, or any solvent, mixture of solvents, or
solvent system that is capable of dissolving chlorite, or a solvent
that is pharmaceutically acceptable for administration in a
subject. Such a solvent is readily identified by those of skill in
the art. See, e.g., Remington: The Science and Practice of
Pharmacy, Twentieth Edition, Lippincott Williams & Wilkins;
20th edition (Dec. 15, 2000), which is incorporated herein by
reference in its entirety. In some embodiments, the solvent is
water. The resulting chlorite solution can include greater than 90%
pure chlorite ions by weight. In some embodiments, the purity of
chlorite in the solution can be greater than 95% pure chlorite ions
or greater than 99% pure chlorite ions by weight. In some
embodiments, wherein the purified chlorite is dissolved in water to
an approximately 1M concentration, the solution includes a pH,
e.g., of about pH 8.5 to about pH 10.
[0121] Alternatively, the chlorite may be suspended in a suspending
medium, including but not limited to any suspending medium that is
capable of suspending chlorite, or a suspending medium that is
pharmaceutically acceptable for administration in a subject. Such a
suspending medium is readily identified by those of skill in the
art. See, e.g., Remington, cited above.
[0122] Briefly, one method of preparing a formulation comprising
chlorite as disclosed herein can be achieve through the steps of:
(a) concentrating a chlorite solution at a temperature between
60.degree. C. to about 100.degree. C., whereby impurities
precipitate from the solution, (b) removing the impurities from the
concentrated solution by filtration, (c) inducing crystallization
of chlorite from the concentrated solution, (d) harvesting the
resulting chlorite solids by filtration, and (e) dissolving the
chlorite solids in an aqueous solvent. It is envisioned that in
some variations the resulting aqueous formulation of chlorite
comprises a purity of at least 80% chlorite, at least 85% chlorite,
at least 90% chlorite, at least 95% chlorite or at least 99%
chlorite.
[0123] The chlorite may also be emulsified in an emulsification
system, including but not limited to any emulsification system that
is capable of emulsifying chlorite, or an emulsification system
that is pharmaceutically acceptable. Such an emulsification system
is readily identified by those of skill in the art. See, e.g.,
Remington, cited above.
[0124] (a) Good Manufacturing Practice (GMP) Formulations and
Methods
[0125] The chlorite formulations for use with the present invention
can be prepared under a standard relating to manufacture and
quality control of pharmaceutical goods, called GMP (Good
Manufacturing Practice), which has been enacted in many countries.
GMP specifies, because of the importance of pharmaceutical goods
which can decide one's life, quality control measures such as
chemical analysis, to maintain optimum equipment and environments
for manufacturing pharmaceutical goods, and to take care of all
manufacturing practices including manufacture, packaging, display,
and storage of pharmaceutical products and materials. GMP further
refers to the Good Manufacturing Practice Regulations promulgated
by the US Food and Drug Administration (FDA) under the authority of
the Federal Food, Drug, and Cosmetic Act. GMP is also sometimes
referred to as "cGMP". The "c" stands for "current" reminding
manufacturers that they must employ technologies and systems which
are up-to-date in order to comply with the regulation. For example,
systems and equipment used to prevent contamination, mix-ups, and
errors, which were state-of-the-art 20 years ago, may be less than
adequate by current standards.
[0126] Accordingly, GMP is well known to those of skill in the art
in relation to the manufacture of pharmaceutical goods. It is
envisioned that the compositions and methods described herein can
be manufactured and performed under GMP or cGMP.
[0127] (b) Methods of Adjusting the pH of Formulations Sensitive to
pH
[0128] Various methods can be used to adjust the pH of formulations
and pharmaceutical formulations comprising chlorite. It is intended
that the methods described herein can be used to produce the
formulations or pharmaceutical formulations described herein for
use with the present invention. However, the formulations and
pharmaceutical formulations described herein may also be produced
by other methods, and the formulations and pharmaceutical
formulations described herein are not limited to those produced by
the methods described herein.
[0129] Some compounds or formulations are sensitive to high local
acidity or alkalinity, requiring proper methods to adjust the pH of
such compounds or formulations. Preferred pH adjusting agent(s) or
pH adjusting compound(s) are weak acids or weak bases having a pKa
of about 4 to about 9, a pKa of about 5 to about 9, or a pKa of
about 5 to about 8, or a pKa of about 6 to about 7.5. Examples
include, but are not limited to a phosphate buffer having a pKa of
about 4 to about 9 as well known in the field, for example,
monobasic phosphates, or monosodium phosphate and/or disodium
phosphate and lower alkanoic acids, for example, acetic acid or
propionic acid. In some embodiments, the pH of a formulation
sensitive to acidity is lowered to between about 7 and about 11.5
using a pH adjusting compound that does not expose the formulation
to acidity, including but not limited to a high local acidity in
the area around the pH adjusting compound. In some embodiments, the
pH of a formulation sensitive to acidity is lowered to between
about 7 and about 10 using a pH adjusting compound that does not
expose the formulation to acidity, including but not limited to a
high local acidity in the area around the pH adjusting compound. In
some embodiments, the pH of a formulation sensitive to acidity is
lowered to between about 7 and about 9.5 using a pH adjusting
compound that does not expose the formulation to acidity, including
but not limited to a high local acidity in the area around the pH
adjusting compound. In some embodiments, the pH of a formulation
sensitive to acidity is lowered to between about 7 and about 9.0
using a pH adjusting compound that does not expose the formulation
to acidity, including but not limited to a high local acidity in
the area around the pH adjusting compound. In some embodiments, the
pH of a formulation sensitive to acidity is lowered to between
about 7 and about 8.5 using a pH adjusting compound that does not
expose the formulation to acidity, including but not limited to a
high local acidity in the area around the pH adjusting compound. In
some embodiments, the pH of a formulation sensitive to acidity is
lowered to between about 7.1 and about 7.7 using a pH adjusting
compound that does not expose the formulation to acidity, including
but not limited to a high local acidity in the area around the pH
adjusting compound.
[0130] "High local acidity," as used herein, refers to the pKa of
one or more molecules local to a chlorite molecule, as opposed to
the overall acidity of a solution as would be measured, for
example, using a pH meter. To determine whether a pH-adjusting
agent will subject chlorite to high local acidity, the pKa of the
pH adjusting agent can be identified using, for example, the CRC
Handbook of Chemistry and Physics (86th Edition, David R. Lide ed.,
CRC Press, 2005).
[0131] Lowering the pH of chlorite formulations has been
challenging because many pH adjusting agents expose compounds or
formulations to high acidity in the local area of the molecules of
the pH-adjusting compound. In the presence of high local acidity,
some amount of non-chlorite compounds are generated, e.g., chlorate
and/or chlorine dioxide. See, e.g., Ullmann's Encyclopedia of
Industrial Chemistry, Vol. A6, Ed. Wolfgang Gerhartz, 5th Ed.
(1986), which is incorporated herein by reference in its entirety.
Such degradation products may not be desired in formulations for
parenteral or systemic administration to physiological systems,
e.g., because they are not inactive in physiological systems. Some
such degradation products result in toxicity, including but not
limited to the toxicities, including but not limited to
non-specific toxicity, described herein.
[0132] Unless the context makes clear, the pH of any of the
formulations or pharmaceutical formulations described herein may be
adjusted using the methods described herein.
[0133] In some variations, the activity of a therapeutic agent,
including but not limited to chlorite, is diminished by exposure to
high local acidity. "Diminished activity," as used herein, refers
to an activity of a therapeutic agent that is qualitatively or
quantitatively inferior to that of the therapeutic agent prior to
the exposure to high local acidity. As one example, a changed
activity that is qualitatively or quantitatively inferior to that
of the therapeutic agent prior to the exposure to high local
acidity would be a lesser efficacy of wound healing, or a lesser
efficacy in treating one or more of the diseases or conditions
described herein. In some variations, the changed activity is any
of at least about 3%, at least about 5%, at least about 10%, at
least about 15%, at least about 20%, or at least about 25% lower
than the activity of the therapeutic agent prior to the exposure to
high local acidity. In some variations, the changed activity is at
least about 5% lower than the activity of the therapeutic agent
prior to the exposure to high local acidity.
[0134] In some embodiments, the pH of a chlorite formulation is
adjusted to any one or more of the pH levels described in the
formulations section or elsewhere herein. In some embodiments, the
pH of a chlorite formulation described between about 7 and about
11.5. In some embodiments, the method comprises lowering the pH of
a formulation comprising chlorite to any of between about between
about 7 and about 11; between about 7 and about 10.5; between about
7 and about 10; between about 7 and about 9.5; between about 7 and
about 9; between about 7 and about 8.5; between about 7 and about
8.0; between about 7 and about 7.5; between about 7.5 and about 8;
between about 7.5 and about 8.5; between about 7 and about 8;
between about 7.1 and about 7.7; between about 7.2 and about 7.6;
between about 7.3 and about 7.5; between about 8 and about 9;
between about 8 and about 8.5; between about 8.5 and about 9; about
7.0; about 7.1; about 7.2; about 7.3; about 7.4; about 7.5; about
7.6; about 7.7; about 7.8; about 7.9; about 8.0; about 8.1; about
8.2; about 8.3; about 8.4; about 8.5; about 8.6; about 8.7; about
8.8; or about 8.9 using a pH adjusting agent that does not expose
the chlorite to a high local acidity. In some embodiments, the
method comprises lowering the pH of a formulation comprising
chlorite to between about 7 and about 8.5. In some embodiments, the
method comprises lowering the pH of a formulation comprising
chlorite to between about 7 and about 8.0. In some embodiments, the
method comprises lowering the pH of a formulation comprising
chlorite to between about 7.1 and about 7.7. In some embodiments,
the method comprises lowering the pH of a formulation comprising
chlorite to about 7.4.
[0135] In one nonlimiting example, the pH of a mixture comprising
chlorite is adjusted using a pH adjusting agent that does not
subject the chlorite to a local pH of below 7 when exposed to the
mixture comprising chlorite. In some embodiments, the pH adjusting
agent is monosodium phosphate, disodium phosphate, or a mixture
thereof. In some embodiments, monosodium phosphate and/or disodium
phosphate is used as a solid or in solution. In some embodiments,
the pH adjusting agent is acetic acid.
[0136] In some embodiments, the pH of chlorite is adjusted by
adding chlorite or an aqueous mixture comprising chlorite to a
solution containing buffer. In some embodiments, the pH of chlorite
is adjusted by adding chlorite or an aqueous mixture comprising
chlorite to a solution of a phosphate buffer.
[0137] In some variations, one or more pH-adjusting agents are used
to adjust the pH of a chlorite solution or mixture, and the
resulting solution or mixture is analyzed for the presence of
degradation products of chlorite, including but not limited to
degradation products generated by high local acidity. In some
variations, pH-adjusting agents such as acetic acid, monosodium
phosphate, and/or disodium phosphate are used to adjust the pH of a
chlorite solution or mixture, and the resulting solution or mixture
is analyzed for the presence of chlorate or chlorine dioxide.
[0138] In some embodiments, the resulting solution or mixture is
analyzed for degradation products using well known analytical
methods such as HPLC, mass spectrometry, etc. In some embodiments,
the resulting solution or mixture is analyzed for degradation
products using a toxicity assay, including well-known toxicity
assays. In some embodiments, the resulting solution or mixture is
analyzed for impurities using a non-specific toxicity assay.
[0139] In some embodiments, the pH of a chlorite formulation is
adjusted after a chlorite purification step.
[0140] In some embodiments, the pH of a chlorite formulation is
adjusted to between about 7 and about 11.5 without the generation
of chlorite degradation products that are a result of high local
acidity. In some embodiments, the pH of a chlorite formulation is
adjusted to between about 7 and about 8.0 without the generation of
chlorite degradation products that are a result of high local
acidity. In some embodiments, the pH of the chlorite formulation is
adjusted to any of between about 7 and about 11; between about 7
and about 10.5; between about 7 and about 10; between about 7 and
about 9.5; between about 7 and about 9; between about 7 and about
8.5; between about 7 and about 8; between about 7 and about 7.5;
between about 7.5 and about 8; between about 7.5 and about 8.5;
between about 7 and about 8; between about 8 and about 9; between
about 8 and about 8.5; or between about 8.5 and about 9 without the
generation of chlorite degradation products that are a result of
high local acidity.
[0141] V. Pharmaceutical Formulations
[0142] Unless the context clearly indicates otherwise, any of the
formulations described herein may be used in any of the
pharmaceutical formulations described herein. In a preferred
embodiment, the pharmaceutical composition comprises: (a) chlorite;
(b) a pH adjusting agent; and (c) a pharmaceutically acceptable
excipient. In some embodiments, the pH adjusting agent comprises
monosodium phosphate and/or disodium phosphate. In some
embodiments, the pH of the composition is between about 7.1 and
about 7.7, e.g., 7.4. The formulations can have low levels of
harmful chlorate, e.g., the weight ratio of chlorite:chlorate can
be greater than 100:1.5. In some embodiments, such formulations are
formulated to be administered intravenously.
[0143] In some embodiments, the pharmaceutical formulations
described herein are suitable for administration to a subject. By
"suitable for administration to a subject" is meant that the
pharmaceutical formulation, when obtained from a newly opened
bottle and administered via the desired route, causes no greater
than a clinically acceptable level of deleterious side effects.
[0144] In some embodiments, the formulations or pharmaceutical
formulations described herein further comprise a saline solution. A
saline solution, as used herein, refers to a physiologically
acceptable solution with a physiologically acceptable level of
sodium chloride. In some embodiments, the saline solution is
isotonic.
[0145] The chlorite formulations for use with the present invention
are pharmaceutically acceptable chlorite formulations comprising
one or more pharmaceutically acceptable excipients. Excipients, as
used herein, refer to any non-chlorite, non-water, or non-saline
element of a pharmaceutical formulation. Excipients include but are
not limited to carriers, adjuvants, diluents, stabilizers, wetting
agents, emulsifiers, buffers, preservatives, flavorings, inactive
ingredients, gel formulations, erodible and non-erodible polymers,
microspheres, liposomes, etc., including combinations of the
foregoing, known to skilled artisans and described further herein.
In some embodiments, the percent by weight of the excipient per the
total volume of the formulation or pharmaceutical formulation is no
greater than any of about 10%, about 9%, about 8%, about 7%, about
6%, about 5%, about 4%, about 3%, about 2%, about 1%, about 0.5%,
about 0.4%, about 0.3%, about 0.2%, about 0.1%, or about 0.05%. In
some embodiments, the percent by weight of the excipient per the
total volume of the formulation or pharmaceutical formulation is no
greater than about 1%. In some embodiments, the percent by weight
of the excipient per the total volume of the formulation or
pharmaceutical formulation is no greater than about 3%.
[0146] Below is a non-limiting and non-exhaustive list of
excipients that are commonly used in the pharmaceutical arts. These
excipients are commonly used in various types of formulations,
including those formulated for intravenous, oral, intramuscular, or
parenteral administration. Given the reactivity of chlorite, it is
likely that some of the excipients listed below are inappropriate
for a given pharmaceutical formulation. Whether or not a particular
excipient is inappropriate for a given pharmaceutical formulation
may depend upon the amount of the excipient being added to the
pharmaceutical formulation. Before adding one or more of any
excipient, including but not limited to the excipients described
herein, to a pharmaceutical formulation of chlorite, it is
important to consider the reactivity of the excipient with
chlorite. Some organic molecules that are commonly used as
excipients react with chlorite in such a way that the excipient is
changed, including but not limited to a change that results in
increased toxicity of the pharmaceutical formulation prior to
exposure of the excipient to chlorite. In some embodiments, the
pharmaceutical formulations described herein comprise one or more
pharmaceutically acceptable excipients that do not react with
chlorite. Preferably, the pharmaceutical formulations described
herein comprise one or more pharmaceutically acceptable excipients
that do not diminish the therapeutic effect of the pharmaceutical
formulation relative to prior to exposure to the excipient.
[0147] In some embodiments, the chlorite formulations described
herein comprise one or more pharmaceutically acceptable excipients
that do not generate one or more of the deleterious non-chlorite
elements of other commercially available chlorite formulations. In
some embodiments, the chlorite formulations described herein
comprise an excipient, and are substantially free of one or more of
the deleterious non-chlorite elements of other commercially
available chlorite formulations. In some embodiments, the chlorite
formulations described herein comprise an excipient, and are
substantially free of one or more of the degradation products or
impurities of other commercially available chlorite formulations as
described herein.
[0148] In some embodiments, the chlorite formulation comprises a
stabilizer. Stabilizers include but are not limited to agents that
will do any of (1) improve the compatibility of excipients with a
container, including a glass bottle or an encapsulating materials
such as gelatin, (2) improve the stability of chlorite (e.g.,
prevent degradation), or (3) improve formulation stability.
[0149] Stabilizers may be selected from, for example, fatty acids,
fatty alcohols, alcohols, long chain fatty acid esters, long chain
ethers, hydrophilic derivatives of fatty acids, polyvinyl
pyrrolidones, polyvinyl ethers, polyvinyl alcohols, hydrocarbons,
hydrophobic polymers, moisture-absorbing polymers, and combinations
thereof. Amide analogues of stabilizers can also be used. The
chosen stabilizer may change the hydrophobicity of the formulation
(e.g., oleic acid, waxes), or improve the mixing of various
components in the formulation (e.g., ethanol), control the moisture
level in the formula (e.g., PVP or polyvinyl pyrrolidone), control
the mobility of the phase (substances with melting points higher
than room temperature such as long chain fatty acids, alcohols,
esters, ethers, amides etc. or mixtures thereof; waxes), and/or
improve the compatibility of the formula with encapsulating
materials (e.g., oleic acid or wax). Some of these stabilizers may
be used as solvents/co-solvents (e.g., ethanol). Stabilizers may be
present in sufficient amount to inhibit chlorite's degradation.
[0150] The formulations described herein may contain one or more of
a gelling agent or a release modifying agent.
[0151] The formulations described herein may contain one or more
adjuvants appropriate for the indicated route of administration.
Again, prior to the addition of any excipient to the formulations
described herein, the reactivity of chlorite should be considered
with respect to whether the resulting pharmaceutical formulation
will be appropriate for administration via the desired route of
administration. Adjuvants with which the therapeutic agent may be
admixed with include but are not limited to lactose, sucrose,
starch powder, cellulose esters of alkanoic acids, stearic acid,
talc, magnesium stearate, magnesium oxide, sodium and calcium salts
of phosphoric and sulphuric acids, acacia, gelatin, sodium
alginate, polyvinylpyrrolidine, and/or polyvinyl alcohol. When a
solubilized formulation is required the therapeutic agent may be in
a solvent including but not limited to polyethylene glycol of
various molecular weights, propylene glycol of various molecular
weights, carboxymethyl cellulose colloidal solutions, methanol,
ethanol, DMSO, corn oil, peanut oil, cottonseed oil, sesame oil,
tragacanth gum, and/or various buffers. Other adjuvants and modes
of administration are well known in the pharmaceutical art and may
be used in the practice of the methods and formulations described
herein. The carrier or diluent may include time delay material,
such as glyceryl monostearate or glyceryl distearate alone or with
a wax, or other materials well known in the art. The formulations
for use as described herein may also include gel formulations,
erodible and non-erodible polymers, microspheres, and
liposomes.
[0152] Additives and diluents normally utilized in the
pharmaceutical arts can optionally be added to the pharmaceutical
composition and the liquid formulation. These include thickening,
granulating, dispersing, flavoring, sweetening, coloring, and
stabilizing agents, including pH stabilizers, other excipients,
anti-oxidants (e.g., tocopherol, BHA, BHT, TBHQ, tocopherol
acetate, ascorbyl palmitate, ascorbic acid propyl gallate, and the
like), preservatives (e.g., parabens), and the like. Exemplary
preservatives include, but are not limited to, benzylalcohol,
ethylalcohol, benzalkonium chloride, phenol, chlorobutanol, and the
like. Some antioxidants provide oxygen or peroxide inhibiting
agents and may be used in the formulations described herein,
including but not limited to, butylated hydroxytoluene,
butylhydroxyanisole, propyl gallate, ascorbic acid palmitate,
a-tocopherol, and the like. Thickening agents, such as lecithin,
hydroxypropylcellulose, aluminum stearate, and the like, may be
used if desired, for example to improve one or more qualities of
the formulation, such as the texture.
[0153] In some variations, the chlorite formulations for use with
the invention are sterile. Sterilization can be by any method that
is compatible with chlorite. In some embodiments, sterilization is
via a method that does not generate a substantial amount of a
degradation product of chlorite. In some embodiments, sterilization
is via a method that does not cause a structural change in
chlorite. In some embodiments, the formulations described herein
are sterile pharmaceutical formulations for parenteral or
intravenous administration. In some embodiments, the chlorite
formulations described herein are sterile filtered, for example,
through a sterile 0.22 micron filter.
[0154] In some embodiments, the formulations or pharmaceutical
formulations are sterile-filterable. In some embodiments, the
chlorite formulations described herein are formulated for
administration by one or more of the routes of administration
described herein. A formulation that is "formulated for
administration" by a specified route of administration, as used
herein, is a formulation that does not include pharmaceutical
excipients that are considered inappropriate for the route of
administration by those of skill in the relevant art. As one
example, a formulation that is suitable for intravenous
administration would not include a toothpaste excipient or carrier
intended for topical administration, where the excipient or carrier
is considered inappropriate for the specified route of
administration by those of skill in the relevant art.
[0155] Chlorite-containing agent in any form disclosed herein can
be provided in any suitable formulation, which can be selected
according to the desired route of administration as disclosed
herein. In one embodiment, the formulation of the drug product
comprises purified sodium chlorite which may include a certain
amount of water content, buffer such as sodium phophate dibasic,
and sterile water for injection (USP) as a vehicle. In one
embodiment, the amount of purified sodium chlorite is about 5.6
mg/mL (including a batch factor to reflect the water content of the
batch), the amount of sodium phosphate dibasic is 0.107 mg/mL, and
sterile water to bring the volume up to 1 mL. In certain
embodiments, a formulation according to the invention consists
essentially of purified sodium chlorite, buffer, and sterile water
for injection (USP) as the vehicle. In certain embodiments, the
formulated drug product is stable for up to 3 months at 25.degree.
C./60% relative humidity and/or 40.degree. C./75% relative humidity
conditions.
[0156] U.S. Pat. No. 4,725,437 describes an aqueous solution of a
chemically stabilized chlorite matrix suitable for intravenous
administration in a dosed amount of about 6.2.times.10.sup.-6 mole
of ClO.sub.2 to 9.3.times.10.sup.-5 mole of ClO.sub.2 per kg of
body weight in humans and non-human animals. The solution contains
the chlorite matrix in a concentration of about 12 to 72 micromol
of ClO.sub.2 per ml. Further chlorite formulations are described in
U.S. Pat. Nos. 4,507,825, and 4,725,437, which are herein
incorporated by reference in their entireties.
[0157] In some embodiments, the present invention provides methods
of treating disease such as type II diabetes or diabetes related
diseases or complications comprising administering an effective
amount of TCDO in a subject. Formulations of TCDO are provided in
this application. In one example, the TCDO formulation is WF10.
WF10 is also known as Oxoferin.RTM. and is available commercially.
In another example, the chlorite formulation contains chlorite.
Other formulations of TCDO or chlorite are encompassed within the
scope of the present invention. Alternatively, in some embodiments
TCDO and/or WF10 can be excluded in part or in whole.
[0158] Chlorite-containing compositions, such as TCDO, can be
formulated for parenteral or enteral administration, generally
parenteral administration. Accordingly, formulations of chlorite,
or chlorite-containing agents such as TCDO and WF10, are suitable
for parenteral, topical or transdermal administration, usually
intravenous, intramuscular, or subcutaneous administration, and may
be suitable for administration by bolus injection, sustained
release (including controlled release), infusion, and the like.
More details on the route of administration are disclosed herein
below. In some embodiments, the administration of the chlorite
containing agents is by infusion e.g., by subcutaneous or
intravenous infusion, or in the form of suppositories.
[0159] VI. Administration and Dosing of Chlorite or Chlorite
Containing Agents
[0160] Unless the context indicates otherwise, all of the
formulations and pharmaceutical formulations described herein may
be administered by any of systemic, parenteral (e.g.,
intramuscular, intraperitoneal, intravenous, ICV, intracisternal
injection or infusion, subcutaneous injection, or implant), by
inhalation spray, nebulized or aerosolized using aerosol
propellants, nasal, vaginal, rectal, sublingual, urethral (e.g.,
urethral suppository), by infusion, intraarterial, intrathecal,
intrabronchial, subcutaneous, intradermal, intravenous,
intracervical, intraabdominal, intracranial, intrapulmonary,
intrathoracic, intratracheal, nasal routes, oral administration
that delivers the therapeutic agent systemically, drug delivery
device, or by a dermal patch that delivers the therapeutic agent
systemically, transdermally or transbuccally. In some variations,
the formulation is formulated for other than oral or transbuccal
administration.
[0161] In some variations, the formulations described herein are
not administered topically.
[0162] In some embodiments, the formulations, pharmaceutical
formulations, and methods of administration and treatment described
herein are suitable for use in any warm- or cold-blooded animal In
some embodiments, the formulations, pharmaceutical formulations,
and methods of administration and treatment described herein are
suitable for use in a mammal, including in the veterinary context,
including domestic pets (such as cats, dogs, rabbits, birds,
horses, etc.) and agricultural animals (such as bovine, ovine,
fowl, etc.). In some variations, the formulations, pharmaceutical
formulations, and methods of administration and treatment described
herein are suitable for use in primates, including but not limited
to humans.
[0163] Chlorite formulations are generally dosed in vivo
corresponding to the body weight of the subject. Due to the
continuous breakdown of the active agent in the blood, the agent is
normally administered at regular intervals. Those of skill in the
art will readily appreciate that actual dosages and regimen will
vary as a function of the agent, formulation, the severity of the
symptoms, the susceptibility of the subject to treatment and/or
side effects, and the like. Dosages are readily and routinely
determined by those of skill in the art by a variety of means.
[0164] Exemplary doses of chlorite-containing formulations can vary
between about 0.1 ml/kg to about 1.5 ml/kg, preferably about 0.5
ml/kg of body weight and at a concentration of about 40 to about 80
mmol ClO.sub.2.sup.- per liter, usually about 60 mMol ClO.sub.2 per
liter, respectively. In the case of TCDO, in some embodiments, WF10
is administered intravenously to patients with diabetes or a
diabetes related disease or complication at a maximum dose of
approximately 0.5 ml/kg. Other suitable doses may be approximately
0.25 ml/kg.
[0165] The regimen of administration e.g. dose combined with
frequency of administration will generally involve administration
in an amount and at a frequency to provide a desired effect, e.g.
administration of an amount effective to provide for improvement in
one or more symptoms of a patient suffering from diabetes or a
diabetes related disease or complication, such as a cardiovascular
disease, a metabolic disease such as metabolic syndrome, and
macular degeneration symptoms. For example, chlorite or a
chlorite-containing agent can be administered for 2, 3, 4, 5, 6, 7,
8, 9, 10 or more consecutive days, which administration period may
be reinitiated after 1, 2, 3 or more weeks following the last
dose.
[0166] Chlorite according to the invention can be administered on a
daily basis. In some embodiments, chlorite is administered on a
daily basis at a dose of about 0.2 mg/kg/day of chlorite to about
3.3 mg/kg/day of chlorite. In some embodiments, chlorite is
administered on a daily basis at a dose of about 0.2 mg/kg/day of
chlorite per day, about 0.4 mg/kg/day of chlorite per day,about 0.5
mg/kg/day of chlorite, about 0.6 mg/kg/day of chlorite, about 0.7
mg/kg/day of chlorite, about 0.8 mg/kg/day of chlorite, about 0.9
mg/kg/day of chlorite, about 1.0 mg/kg/day of chlorite, about 1.1
mg/kg/day of chlorite, about 1.2 mg/kg/day of chlorite, about 1.3
mg/kg/day of chlorite, about 1.4 mg/kg/day of chlorite, about 1.5
mg/kg/day of chlorite, about 1.6 mg/kg/day of chlorite, about 1.7
mg/kg/day of chlorite, about 1.8 mg/kg/day of chlorite, about 1.9
mg/kg/day of chlorite, about 2.0 mg/kg/day of chlorite, about 2.1
mg/kg/day of chlorite, about 2.2 mg/kg/day of chlorite, about 2.3
mg/kg/day of chlorite, about 2.4 mg/kg/day of chlorite, about 2.5
mg/kg/day of chlorite, about 2.6 mg/kg/day of chlorite, about 2.7
mg/kg/day of chlorite, about 2.8 mg/kg/day of chlorite, about 2.9
mg/kg/day of chlorite, about 3.0 mg/kg/day of chlorite, about 3.1
mg/kg/day of chlorite, about 3.2 mg/kg/day of chlorite, about 3.3
mg/kg/day of chlorite, about 3.4 mg/kg/day of chlorite, or about
3.5 mg/kg/day of chlorite.
[0167] In some embodiments, the pharmaceutical composition used in
the methods of the invention can be further administered in a
cycle. An exemplary cycle consists of: a) a first period of time
wherein the pharmaceutical composition is administered at a first
dose for a first number of times; and b) a second period of time
wherein the pharmaceutical composition is administered at a second
dose for a second number of times. In some embodiments, the first
period of time is about one week, the first number of times is
about five, the second period of time is about two weeks, and the
second number of times is zero. In other embodiments, the first
period of time is about one week, the first number of times is
about three, the second period of time is about one week, and the
second number of times is zero. The first dose can be about 0.4
mg/kg/day of chlorite to about 3.3 mg/kg/day of chlorite. For
example, the first dose can be about 2.1 mg/kg/day of chlorite. The
cycle can be performed multiple times, e.g., about 2, 3, 4, 5, 6,
7, 8, 9, 10 or 10 or more times. In some embodiments, the cycle is
performed about 2-4 times.
[0168] In some embodiments, the dosing schedule consists of periods
of administration alternating with periods of non-administration.
For example, chlorite might be administered in a three week cycle,
comprising dosing chlorite up to 5 times in a week followed by two
weeks without treatment. The cycle could be repeated as necessary
to achieve the desired result. In another embodiment, chlorite is
administered in a two week cycle, e.g., up to 3 times in a week
followed by a week without administration. In some embodiments, a
total of 2-4 cycles are performed. In an exemplary embodiment, the
dosing regimen comprises administration of 2.1 mg/kg/day of
chlorite for a total of 2-4 three week cycles.
B. Immunomodulating Agents
[0169] In one aspect, the present invention provides a method of
treating a macrophage related disease comprising administering to a
subject in need thereof an effective amount of an immunomodulating
agent. In another aspect, the present invention provides a method
of modulating macrophage accumulation or activation comprising
administering to a subject in need thereof an effective amount of
an immunomodulating agent. In some embodiments, the
immunomodulating agent is an immunosuppressor and downregulates the
immune response. In some embodiments, the agent has
immunostimulatory effects. The immunosuppressor can act directly
and/or indirectly on activated macrophage. In some embodiments, the
immunosuppressor acts directly on macrophage, e.g., by inhibiting
macrophage activity. In some embodiments, the immunosuppressor
disrupts immune cell signaling, e.g., by interfering with the
production or activity of cytokines produced by immune cells such
as T-cell, B-cells or macrophage. Any desirable combination of
immunmodulatory effects may be observed. In one embodiment, the
present invention provides for methods of treating non-MS
neuroinflammation with an immunomodulatory agent capable of
blocking migration of monocytes or activated macrophages.
[0170] In some embodiments, the present invention provides for
lower dosages of the immunomodulating agents than typically used,
e.g., when used as monotherapy. For example, the immunomodulating
agent can be used in combination with an oxidative agent, including
but not limited to chlorite or a chlorite-containing agent, thereby
allowing a lower dosage of the immunomodulating agent to achieve
the desired therapeutic effect. In such instances, the oxidative
agent potentiates the effect of the immunomodulating agent.
Alternatively, the oxidative agent can be administered to
ameliorate or reduce toxicity or other side effects of the
immunomodulating agent, thereby allowing for a higher dosage of he
immunomodulating agent than would otherwise be tolerated during
monotherapy with the immunomodulating agent. For example, in
certain embodiments, combination therapy with the oxidative agent
allows for administration of an amount of immunomodulating agent
that might otherwise result in an unacceptable level of
opportunistic infections.
[0171] Immunosuppressive drugs or immunosuppressive agents inhibit
or prevent activity of the immune system. Immunosuppressive therapy
has a number of uses, including but not limited to preventing the
rejection of transplanted organs and tissues (e.g., bone marrow,
heart, kidney, liver), treating autoimmune diseases or diseases
that are most likely of autoimmune origin (e.g., rheumatoid
arthritis, multiple sclerosis, myasthenia gravis, systemic lupus
erythematosus, Crohn's disease, pemphigus, and ulcerative colitis),
and also treating some other non-autoimmune inflammatory diseases
(e.g., long term allergic asthma control).
[0172] Immunosuppressants have certain side-effects and risks. For
example, in some cases, the immune system is less able to resist
infections and the spread of malignant cells. Other side-effects
include hypertension, dyslipidemia, hyperglycemia, peptic ulcers,
liver, and kidney injury. Immunosuppressive agent can also interact
with and affect the metabolism and action of other therapeutic
agents. In some embodiments, the present invention provides for
lower dosages of the immunosuppressive agents than typically used,
e.g., for transplant rejection. For example, the immunosuppressive
agent can be used in combination with an oxidative agent, including
but not limited to chlorite or a chlorite-containing agent, thereby
allowing lower dosage of the immunosuppressant to achieve the
desired therapeutic effect. In such instances, the oxidative agent
potentiates the effect of the immunosuppressant.
[0173] Immunosuppressive drugs can generally be classified as
glucocorticoids, cytostatics, antibodies, drugs acting on
immunophilins, or other drugs.
[0174] Glucocorticoids are a class of steroid hormones that bind to
and activate the glucocorticoid receptor. The activated
glucocorticoid receptor complex in turn up-regulates the expression
of anti-inflammatory proteins in the nucleus (a process known as
transactivation) and represses the expression of pro-inflammatory
proteins in the cytosol by preventing the translocation of other
transcription factors from the cytosol into the nucleus
(transrepression). Glucocorticoids suppress cell-mediated immunity
by inhibiting genes that code for the cytokines IL-1, IL-2, IL-3,
IL-4, IL-5, IL-6, IL-8, and TNF-.gamma.. They also suppress IL-2
and IL-2 receptor production by B-cells, thereby reducing B cell
clonal expansion and antibody production.
[0175] Glucocorticoids induce the synthesis of lipocortin-1
(annexin-1), leading to diminished pro-inflammatory eicosanoid
production. The expression of cyclooxygenases COX-1 and COX-2 is
also suppressed, further inhibiting inflammation. Glucocorticoids
also stimulate the lipocortin-1 escaping to the extracellular
space, where it binds to the leukocyte membrane receptors and
inhibits various inflammatory events: epithelial adhesion,
emigration, chemotaxis, phagocytosis, respiratory burst, and the
release of various inflammatory mediators (lysosomal enzymes,
cytokines, tissue plasminogen activator, chemokines, etc.) from
neutrophils, macrophages, and mastocytes. Glucocorticoids
down-regulate the expression of Fc receptors on macrophages,
thereby decreased phagocytosis of opsonised cells.
[0176] Glucocorticoids for use with the present invention include
without limitation hydrocortisone (cortisol), cortisone acetate,
prednisone, prednisolone, methylprednisolone, dexamethasone,
betamethasone, triamcinolone, beclometasone, fludrocortisone
acetate, deoxycorticosterone acetate (doca) and aldosterone.
[0177] Cytostatics inhibit cell division. Used at lower levels that
those used to treat cancer, cytostatics can affect the
proliferation of immune cells, e.g., B-cells and T-cells.
Alkylating agents used as immunosuppressants include nitrogen
mustards (cyclophosphamide), nitrosoureas, platinum compounds, and
others. Antimetabolites interfere with the synthesis of nucleic
acids. These include purine synthesis inhibitors azathioprine,
mercaptopurine and mycophenolic acid; pyrimidine synthesis
inhibitors leflunomide and teriflunomide; and the folic acid analog
methotrexate. Cytotoxic antibiotics include without limitation
dactinomycin, anthracyclines, mitomycin C, bleomycin, and
mithramycin.
[0178] Antibodies can also be used as immune modulators. An
"antigen" is an entity to which an antibody specifically binds. The
term "antibody" or "immunoglobulin" is used to include intact
antibodies and binding fragments thereof. Typically, fragments
compete with the intact antibody from which they were derived for
specific binding to an antigen fragment including separate heavy
chains, light chains Fab, Fab', F(ab').sub.2, F(ab)c, and Fv.
Fragments may be produced by recombinant DNA techniques, or by
enzymatic or chemical separation of intact immunoglobulins. The
term "antibody" also includes one or more immunoglobulin chains
that are chemically conjugated to, or expressed as, fusion proteins
with other proteins. The term "antibody" also includes bispecific
antibody. A bispecific or bifunctional antibody is an artificial
hybrid antibody having two different heavy/light chain pairs and
two different binding sites. Bispecific antibodies can be produced
by a variety of methods including fusion of hybridomas or linking
of Fab' fragments (See, e.g., Songsivilai and Lachmann, Clin. Exp.
Immunol , 79:315-321 (1990); Kostelny et al., J. Immunol,
148:1547-53 (1992)). An antibody or fragment thereof may be labeled
with a detectable marker or conjugated to a second molecule, such
as a therapeutic agent (e.g., a cytotoxic agent) thereby resulting
in an immunoconjugate. For example, the therapeutic agent includes,
but is not limited to, an anti-tumor drug, a toxin, a radioactive
agent, a cytokine, a second antibody or an enzyme. The
immunoconjugate also includes an embodiment wherein the antibody of
the invention is linked to an enzyme that converts a prodrug into a
cytotoxic drug.
[0179] Heterologous polyclonal antibodies can affect all
lymphocytes and cause general immunosuppression. Anti-thymocyte
globulin and anti-lymphocyte globulin comprise horse and rabbit
polyclonal antibodies against human T cells. Preparations available
to the market in Atgam.RTM., obtained from horse serum, and
Thymoglobuline.RTM., obtained from rabbit serum. Because of a high
immunogenicity of heterologous polyclonal antibodies, almost all
patients have an acute reaction to the treatment. Monoclonal
antibodies are directed towards defined target antigens and cause
fewer side-effects. Common treatments include IL-2 receptor-
(CD25-) and CD3-directed antibodies. OKT3 (also called muromonab)
is a murine anti-CD3 monoclonal antibody that prevents T-cell
activation and proliferation by binding the T-cell receptor complex
present on all differentiated T cells. It is one of the most potent
immunosuppressive agents. CD3 is also targeted by the monoclonal
antibodies otelixizumab, teplizumab, and visilizumab.
[0180] CD4 (cluster of differentiation 4) is a glycoprotein
expressed on the surface of T helper cells, regulatory T cells,
monocytes, macrophages, and dendritic cells. Antibodies against CD4
include clenoliximab, keliximab, and zanolimumab.
[0181] Efalizumab (Raptiva.RTM.) is a recombinant humanized
monoclonal antibody that binds to CD11a and acts as an
immunosuppressant by inhibiting white blood cell migration out of
blood vessels into tissues.
[0182] Erlizumab and rovelizumab are humanized monoclonal
antibodies that bind CD18. They are intended to suppress white
blood cells that become overly active, e.g., during shock.
[0183] CD20 is a non-glycosylated phosphoprotein expressed on the
surface of mature B-cells. Anti-CD20 antibodies include afutuzumab,
ocrelizumab, ofatumumab and pascolizumab. Rituximab is a chimeric
monoclonal antibody against CD20, which is primarily found on the
surface of B cells. Rituximab thereby destroys both normal and
malignant B cells that express CD20.
[0184] Lumiliximab is a chimeric monoclonal antibody that targets
against CD23 to acts as an immunomodulator.
[0185] CD40 is a costimulatory protein found on antigen presenting
cells and is required for their activation. In the macrophage, the
primary signal for activation is IFN-.gamma. from Th1 type CD4 T
cells. The secondary signal is CD40 ligand (CD40L or CD154) on the
T cell which binds CD40 on the macrophage cell surface. As a
result, the macrophage expresses more CD40 and TNF receptors on its
surface which helps increase the level of activation. Activation
results in the induction of potent microbicidal substances in the
macrophage, including reactive oxygen species and nitric oxide,
which can destroy ingested microbes. Teneliximab and Toralizumab
are monoclonal antibodies against CD40. Ruplizumab is an anti-CD40L
monoclonal antibody.
[0186] Aselizumab is a monoclonal antibody that binds to CD62L
(L-selectin) a cell adhesion molecule found on leukocytes.
[0187] Galiximab is a chimeric monoclonal antibody that binds CD80.
CD80 is found on activated B cells and monocytes and provides a
costimulatory signal necessary for T cell activation and survival.
It is also known as B7.1.
[0188] Gavilimomab is a mouse monoclonal antibody (also known as
ABX-CBL) that binds antigen CD147 (basigin), which is expressed by
many cell types, including epithelial cells, endothelial cells and
leukocytes.
[0189] Belimumab is a fully human monoclonal antibody that
specifically recognizes and inhibits the biological activity of
B-Lymphocyte stimulator (BLyS), also known as B cell activation
factor of the TNF family (BAFF). BLyS is secreted, sometimes under
the influence of interferon-gamma, by a variety of cells, including
monocytes and macrophages.
[0190] CTLA4 (Cytotoxic T-Lymphocyte Antigen 4) also known as CD152
(Cluster of differentiation 152) is expressed on the surface of
Helper T cells and transmits an inhibitory signal to T cells. The
CTLA4-Ig fusion proteins abatacept and belatacept act as
immunosuppressants. In contrast, ipilimumab and tremelimumab are
monoclonal antibodies that bind CTLA4 and to stimulate immune
response.
[0191] Bertilimumab is a human monoclonal antibody that binds to
eotaxin 1, a chemokine that selectively recruits eosinophils by
inducing their chemotaxis.
[0192] Natalizumab is a humanized monoclonal antibody against the
cellular adhesion molecule .alpha.4-integrin. Natalizumab is
believed to reduce the ability of inflammatory immune cells to
attach to and pass through the cell layers lining the intestines
and blood-brain barrier. Natalizumab may be available as
Tysabri.RTM..
[0193] Tocilizumab is a humanized monoclonal antibody against IL-6R
(interleukin-6 receptor). IL-6 is an interleukin that acts as both
a pro-inflammatory and anti-inflammatory cytokine. It is secreted
by T cells and macrophages to stimulate immune response to trauma,
especially burns or other tissue damage leading to
inflammation.
[0194] Lymphocyte function-associated antigen 1 (LFA-1) is found on
T-cells, B-cells, macrophages and neutrophils and is involved in
recruitment to the site of infection. Odulimomab is a mouse
monoclonal antibody that binds LFA-1.
[0195] Interleukin-2 (IL-2) plays a role in the clone expansion and
survival of activated T lymphocytes. Basiliximab (Simulect.RTM.),
daclizumab (Zenapax.RTM.) and inolimomab are monoclonal antibodies
that bind the IL-2a receptor's a chain (i.e., CD25), thereby
preventing the IL-2 induced clonal expansion of activated
lymphocytes and shortening their survival.
[0196] Zolimomab aritox is a conjugate of a murine monoclonal
antibody with the A chain of the toxin ricin. It is directed
against the CD5 antigen of human lymphocytes.
[0197] Siplizumab (MEDI-507) is a monoclonal antibody with a human
IgG1, kappa directed to CD2. The agent has shown potent
immunomodulatory effects, selectively suppressing the function of T
and NK cells.
[0198] Other antibodies and fusion proteins that act as
immunosuppressants include nerelimomab, faralimomab, atlizumab,
atorolimumab, cedelizumab, dorlimomab aritox, dorlixizumab,
fontolizumab, gantenerumab, gomiliximab, lebrilizumab, maslimomab,
morolimumab, pexelizumab, reslizumab, rovelizumab, talizumab,
telimomab aritox, vapaliximab, vepalimomab, aflibercept, alefacept
and rilonacept.
[0199] Immunophilins are peptidyl-prolyl isomerases that play a
role in immunoregulation and basic cellular processes involving
protein folding and trafficking. Modulators of members of the
immunophilin FKBP/cyclophilin/calcineurin pathway can inhibit
cytokine production and activity, such as that of IL-2. Such agents
include rapamycin, tacrolimus, ciclosporin, pimecrolimus, abetimus
and gusperimus. The mammalian target of rapamycin (mTOR) protein,
also known as FK506 binding protein 12-rapamycin associated protein
1 (FRAP1), is a serine/threonine protein kinase that regulates cell
growth, cell proliferation, cell motility, cell survival, protein
synthesis, and transcription. mTOR inhibitors include rapamycin
(sirolimus) and related drugs, including ridaforolimus, everolimus,
temsirolimus, and zotarolimus.
[0200] Other classes of immunosuppressive agents include
interferons, tumor necrosis factor-alpha (TNF.alpha. or TNF)
modulators, IL-1 receptor antagonists, opiods, mycophenolate and
other small biological agents. Interferon-.beta. (IFN-.beta.)
suppresses the production of Th1 cytokines and the activation of
monocytes. TNF is a cytokine that promotes the inflammatory
response. TNF inhibitors include monoclonal antibody such as
infliximab (Remicade.RTM.), adalimumab (Humira.RTM.), certolizumab
pegol (Cimzia.RTM.), and golimumab (Simponi.TM.), or with a
circulating receptor fusion protein such as etanercept
(Enbrel.RTM.). Small molecule TNF inhibitors include thalidomide,
lenalidomide, pentoxifylline and bupropion. Natural TNF inhibitors
include curcumin and catechin.
[0201] Therapeutic agents are available to modulate other
cytokines. The cytokine IL-1 also plays a role in the inflammatory
response. Therapeutic agents that interfere with signal reception
include the IL-1 receptor antagonist anakinra, a recombinant,
non-glycosylated version of human IL-1. Mepolizumab is a humanized
monoclonal antibody that recognizes interleukin-5 (IL-5) and
interferes with proliferation of eosinophil granulocytes.
Omalizumab is another humanized monoclonal antibody that inhibits
the binding of IgE to the high-affinity IgE receptor Fc.epsilon.RI.
Talizumab also binds IgE. Ustekinumab is a human monoclonal
antibody that recognizes and inhibits Interleukin 12 and
Interleukin 23, which help activiate certain T-cells.
[0202] Opiods can suppress both innate and adaptive immunity. A
decrease in proliferation as well as immune function has been
observed in macrophages and lymphocytes. Mycophenolic acid acts as
a non-competitive, selective, and reversible inhibitor of
Inosine-5'-monophosphate dehydrogenase (IMPDH), which is a key
enzyme in the de novo guanosine nucleotide synthesis. Unlike other
human cell types, lymphocytes B and T are very dependent on this
process. Myriocin is an atypic amino acid that can inhibit the
proliferation of IL-2-dependent cytotoxic T cells. Fingolimod is
chemically similar to the myriocin and prevents egress of
lymphocytes from lymph nodes.
[0203] NF-.kappa.B (nuclear factor kappa-light-chain-enhancer of
activated B cells; NFkB) is a dimeric protein complex that controls
DNA transcription. NF-.kappa.B is a rapid responder to cellular
stimuli such as stress, cytokines, free radicals, ultraviolet
irradiation, oxidized LDL, and bacterial or viral antigens.
NF-.kappa.B plays a key role in regulating the immune response to
infection. Improper regulation of NF-.kappa.B has been linked to
cancer, inflammatory and autoimmune diseases, septic shock, viral
infection, and improper immune development. NF-.kappa.B has also
been implicated in processes of synaptic plasticity and memory.
NF-.kappa.B is one of the most crucial transcription factors
regulating the inflammatory repertoire of macrophages, particularly
their expression of proinflammatory cytokines, costimulatory
molecules, and other activation markers in response to diverse
environmental cues (e.g., stress signals, inflammatory cytokines,
pathogens, and hypoxia).
[0204] Prior to stimuli as described above, NF-.kappa.B dimers are
sequestered in the cytoplasm by a family of inhibitors, called
I.kappa.Bs (Inhibitor of .kappa.B), which are proteins that contain
multiple ankyrin repeat sequences. Activation of the NF-.kappa.B is
initiated by the signal-induced degradation of I.kappa.B proteins
by the proteasome. This signal occurs primarily via activation of a
kinase called the I.kappa.B kinase (IKK). With the degradation of
the I.kappa.B inhibitor, the NF-.kappa.B complex can then enter the
nucleus and activate the expression of specific genes that have
nearby DNA-binding sites for NF-.kappa.B. The activation of these
genes by NF-.kappa.B then leads to the given physiological
response, for example, an inflammatory or immune response, a cell
survival response, or cellular proliferation. NF-.kappa.B also
activates the production of its repressor I.kappa.B.alpha., thereby
acting to inhibit its own synthesis.
[0205] Inhibitors of NF-.kappa.B can act at various steps of the
NF-.kappa.B signaling cascade. Some inhibitors of NF-.kappa.B are
described in U.S. Pat. No. 6,410,516. Other agents include
raloxifene, an oral selective estrogen receptor modulator, and
drotrecogin alfa, a recombinant form of human activated protein C
that has anti-thrombotic, anti-inflammatory, and profibrinolytic
properties. The monoclonal antibody denosumab indirectly inhibits
NF-.kappa.B. The drugs disulfiram, olmesartan and dithiocarbamates
can inhibit the NF-.kappa.B signaling cascade. Other direct or
indirect inhibitors of NF-.kappa.B include but are not limited to
proteasome inhibitors (e.g., bortezomib, MG132, Pro1, NPI-0052),
natural small molecules (curcumin, genistein, resveratrol,
parthenolide), thalidomide, lenalidomide, flavopiridol,
non-steroidal anti-inflammatory drugs (NSAIDs), arsenic trioxide,
dehydroxymethylepoxyquinomycin (DHMEQ),
I3C(indole-3-carbinol)/DIM(di-indolmethane) (I3C/DIM), Bay 11-7082,
luteolin, cell permeable peptides such as SN-50, and gene therapy
such as overexpression of I.kappa.B.alpha.-super repressor and
NF.kappa.B decoy oligodeoxynucleotide (ODN). See, e.g., Melisi D,
Chiao P J. NF-kappa B as a target for cancer therapy. Expert Opin
Ther Targets. 2007 February; 11(2):133-44; Baud V, Karin M. Is
NF-kappaB a good target for cancer therapy? Hopes and pitfalls. Nat
Rev Drug Discov. 2009 January; 8(1):33-40.
[0206] Immunomodulators that directly or indirectly modulate
NF-.kappa.B can be used in the methods of the invention. In some
embodiments, immunomodulators are used that suppress the activity
of NF-.kappa.B. In some embodiments, immunomodulators are used that
promote the activity of NF-.kappa.B.
[0207] Immunosuppressants that suppress the activation of
macrophages, or suppress the activity of activated macrophages can
be used in the methods of the invention. In some embodiments,
agents that suppress the production or activity of stimulatory
cytokines and other immune modulatory signals are used. In some
embodiments, agents that block recruitment of immune cells are
used. The methods of the invention also include the treatment of
macrophage related diseases and disorders with immunomodulating
agents through indirect effects on macrophages. For example,
macrophage migration inhibitory factor (MIF) is a known
pro-inflammatory cytokine. See U.S. Patent Publication No.
2009/0170951 ("Description of the Related Art"). Methods according
to the invention include inhibitors and/or antagonists of MIF, such
as N-acetyl-p-benzoquinone imine (NAPQI),
(S,R)-3-(4-hydroxyphenyl)-4,5-dihydro-5-isoxazole acetic acid
methyl ester (ISO-1),
2-[3-(4-hydroxy-phenyl)-4,5-dihydro-isoxazol-5-yl]-3-phenyl-propanoic
acid and its esters, including the methyl ester,
phenylmethylsulfonyl fluoride (PMSF), Cyc-Oxi-11,
2-oxo-4-phenyl-3-butynoate, AVP-13546, phenylpyruvate,
2-[(4-hydroxybenzylidene)amino]-3-(1H-indol-3-yl)propionic acid
methyl ester, and anti-MIF antibodies. For additional example, see
Garai et al., Curr. Med. Chem. 2009; 16(9): 1091-114, and U.S. Pat.
No. 7,662,843, each of which is hereby incorporated by reference in
its entirety. In various embodiments, immunomodulators according to
the invention may include inhibitors and/or antagonists of monocyte
chemo attractant protein-1 (MCP-1).
[0208] Immunomodulators that directly or indirectly modulate cell
adhesion can be used in the methods of the invention. In some
embodiments, immunomodulators are used that suppress the expression
or activity of cell adhesion proteins. In various embodiments, such
cell adhesion molecules may include IgSF type molecules (N-CAM
(Myelin protein zero), ICAM (1,5), VCAM-1, PE-CAM, L1-CAM),
Integrins (LFA-1, Integrin alphaXbeta2, Macrophage-1 antigen,
VLA-4, Glycoprotein IIb/IIIa), Cadherins (Desmoglein (DSG1, DSG2,
DSG3, DSG4), Desmocollin (DSC1, DSC2, DSC3), T-cadherin,
Protocadherin, CDH1, CDH2, CDH3, CDH4, CDH5, CDH6, CDH8, CDH11,
CDH12, CDH15, CDH16, CDH17), Selectins (E-selectin, L-selectin,
P-selectin), or other types of molecules (Carcinoembryonic antigen,
CD22, CD24, CD44, CD146, CD164).
[0209] In some embodiments, the macrophage related diseases that
can be treated using the methods of the present invention are
inflammatory diseases.
C. Macrophage Activation
[0210] In one aspect, the present invention provides a method of
modulating macrophage accumulation or activation comprising
administering an effective amount of an immunomodulatory agent in
combination with an oxidative agent to a subject in need thereof to
treat macrophage related diseases or related conditions. The
oxidative agent may include, but is not limited to, chlorite.
[0211] Macrophages are released from the bone marrow as immature
monocytes, circulate in the blood stream, and can eventually
migrate into tissues to undergo final differentiation into resident
macrophages. Resident macrophages include Kupffer cells in the
liver, alveolar macrophages in the lung, and osteoclasts in the
bone. Monocytes and macrophages are phagocytes, acting in innate
immunity as well as to help adaptive immunity of vertebrate
animals. Their role is to phagocytose (engulf and then digest)
cellular debris and pathogens either as stationary or mobile cells,
and to stimulate lymphocytes and other immune cells to respond to
the pathogen. They can be identified by specific expression of a
number of proteins including CD14, CD11b, F4/80 (mice)/EMR1
(human), Lysozyme M, MAC-1/MAC-3 and CD68 by flow cytometry or
immunohistochemical staining (Khazen W, et al. 2005 FEBS Lett. 579
(25): 5631-4). When a monocyte enters damaged tissue through the
endothelium of a blood vessel (a process known as the leukocyte
extravasation), it undergoes a series of changes to become a
macrophage. Monocytes are attracted to a damaged site by chemical
substances through chemotaxis, triggered by a range of stimuli
including damaged cells, pathogens and cytokines released by
macrophages already at the site. At some sites such as the testis,
macrophages have been shown to populate the organ through
proliferation. Unlike short-lived neutrophils, macrophages survive
longer in the body up to a maximum of several months.
[0212] Macrophages perform a multitude of functions essential for
tissue remodeling, inflammation, and immunity, including but not
limited to phagocytosis, cytotoxicity, and secretion of a variety
of cytokines, growth factors, lysozymes, proteases, complement
components, coagulation factors, and prostaglandins. One important
role of the macrophage is the removal of necrotic cellular debris
in the lungs. Removing dead cell material is important in chronic
inflammation as the early stages of inflammation are dominated by
neutrophil granulocytes, which are ingested by macrophages if they
come of age. The removal of necrotic tissue is to a greater extent
handled by fixed macrophages, which typically stay at strategic
locations such as the lungs, liver, neural tissue, bone, spleen and
connective tissue, where microbial invasion or accumulation of dust
is likely to occur, ingesting foreign materials such as pathogens,
recruiting additional macrophages if needed. Macrophages can
express paracrine functions within organs that are specific to the
function of that organ. In the testis for example, macrophages have
been shown to be able to interact with Leydig cells by secreting
25-hydroxycholesterol, an oxysterol that can be converted to
testosterone by neighboring Leydig cells. Also, testicular
macrophages may participate in creating an immune privileged
environment in the testis, and in mediating infertility during
inflammation of the testis. A list of different types of
macrophages in tissues is shown in Table 1.
TABLE-US-00001 TABLE 1 Name of cell Location Dust cells/Alveolar
macrophages pulmonary alveolus of lungs Histiocytes connective
tissue Kupffer cells liver Microglia neural tissue Epithelioid
cells granulomas Osteoclasts bone Sinusoidal lining cells spleen
Mesangial cells kidney
[0213] Macrophages as scavengers remove dying cells and other
debris from the body. They are a type of antigen presenting cells
which play a crucial role in initiating an immune response. As
secretory cells, monocytes and macrophages are vital to the
regulation of immune responses and the development of inflammation
as they produce monokines including enzymes, complement proteins,
and regulatory factors such as interleukin-1. Macrophages also
carry receptors for lymphokines for lymphocyte activation important
for killing microbes and tumor cells. After digesting a pathogen, a
macrophage presents the antigen on a MHC class II molecule to the
corresponding helper T cell. Eventually the antigen presentation
results in the production of antibodies that bind to the antigens
of pathogens, leading to phagocytosis or antibody-dependent cell
cytotoxicity by macrophages. The antigen presentation on the
surface of infected macrophages (in the context of MHC class II) in
a lymph node stimulates TH1 (type 1 helper T cells) to proliferate
(mainly due to IL-12 secretion from the macrophage). When a B-cell
in the lymph node recognizes the same unprocessed surface antigen
on the microbe with its surface bound antibody, the antigen is
endocytosed and processed. The processed antigen is then presented
in MHCII on the surface of the B-cell. TH1 receptor that has
proliferated recognizes the antigen-MHCII complex (with
co-stimulatory factors- CD40 and CD40L) and causes the B-cell to
produce antibodies that help opsonisation of the antigen so that
the pathogen can be better cleared by macrophages.
[0214] Macrophages provide yet another line of defense against
tumor cells and somatic cells infected with fungus or parasites.
Once a T cell has recognized its particular antigen on the surface
of an aberrant cell, the T cell becomes an activated effector cell
producing lymphokines including families of interleukins,
chemokines and interferons that further stimulate and activate
macrophages. These activated macrophages can then engulf and digest
affected cells more efficiently. The macrophage does not generate a
response specific for an antigen, but attacks the cells present in
the local area in which it was activated.
[0215] Macrophages also play a role in muscle regeneration. A
previous study has shown macrophage influences on muscle repair of
soleus muscle on mice (Tidball J G, Wehling-Henricks M, 2007, The
Journal of Physiology 578: 327-336). Macrophage depletion also
reduces muscle growth during a growth period.
[0216] I. Classically Activated Macrophages
[0217] In one aspect, the present invention provides a method of
modulating macrophage accumulation or activation comprising
administering an effective amount of an oxidative agent or an
immunomodulatory agent. In some embodiments, the oxidative agent is
chlorite. In some embodiments, the oxidative agent is WF10. In some
embodiments, the immunomodulatory agent is an immunosuppressant. In
some embodiments, the oxidative or immunomodulating agent modulates
the stimulation of macrophages via receptors expressed by
macrophages including but not limited to interferon (IFN)-gamma
receptor, CD14/LPS receptor, MHC II molecule, or interleukin
receptors such as IL-4 and IL-13 receptors. In some embodiments,
the oxidative or immunomodulating agent modulates the release of
chemokines by macrophages. In some embodiments, the oxidative or
immunomodulating agent modulates the release of pro-inflammatory
cytokines such as IL-1, IL-6, and TNF-alpha, or anti-inflammatory
cytokines such as IL-10 and TGF-beta by macrophages. In some
embodiments, the oxidative or immunomodulating agent modulates the
release of proteolytic enzymes by macrophages. In some embodiments,
the oxidative or immunomodulating agent modulates the release of
extracellular matrix (ECM) related molecules by macrophages.
[0218] A model of two major macrophage classes has developed
(Gordon, S. (1999) Fundamental Immunology, 4th Ed., Paul, W. E.,
ed., Lippincott-Raven Publishers, Philidelphia, pp. 533-545; Stein,
M. et al. (1992) J. Exp. Med. 176:287). Classically activated
macrophages typically exhibit a Th1-like phenotype, promoting
inflammation, extracellular matrix (ECM) destruction, and
apoptosis, while alternatively activated macrophages typically
display a Th2-like phenotype, promoting ECM construction, cell
proliferation, and angiogenesis. Although both phenotypes are
important components of both the innate and adaptive immune
systems, the classically activated macrophage tends to elicit
chronic inflammation and tissue injury whereas the alternatively
activated macrophage tends to resolve inflammation and facilitate
wound healing (See reviews: Duffield, J. S. (2003) Clin. Sci.
104:27; Gordon, S. (2003) Nat. Rev. Immunol 3:23; Ma, J. et al.
(2003) Cell. Mol. Life Sci. 60:2334; Mosser, D. M. (2003) J.
Leukoc. Biol. 73:209).
[0219] Differentiation of classically activated macrophages
requires a priming signal in the form of IFN-gamma via the
IFN-gamma R (Dalton, D. K. et al. (1993) Science 259:1739; Huang,
S. et al. (1993) Science 259:1742). When the primed macrophage
subsequently encounters an appropriate stimulus, such as bacterial
LPS, it becomes classically activated. LPS is first bound by
soluble LBP and then by either soluble or membrane-bound CD14. CD14
delivers LPS to the LPS recognition complex (Janeway, C. A. &
R. Medzhitov (2002) Annu. Rev. Immunol 20:197), which consists of
at least TLR410 and MD-2 (Nagai, Y. et al. (2002) Nat. Immunol.
3:667). Pathogens and pathogen components are subsequently taken up
by phagocytosis (Honey, K. & A. Y. Rudensky (2003) Nat. Rev.
Immunol. 3:472) and delivered to lysosomes where they are exposed
to a variety of degradation enzymes including several cathepsin
cysteine proteases. Suitable antigens are processed and loaded onto
MHC class II molecules in late endocytic compartments and
antigen/MHCII complexes as well as co-stimulatory B7 family members
are presented to T cells (Harding, C. V. et al. (2003) Curr. Opin.
Immunol 15:112).
[0220] These events are followed closely by a significant change in
cellular morphology and a dramatic alteration in the secretory
profile of the cell. A variety of chemokines including IL-8/CXCL8,
IP-10/CXCL10, MIP-1 alpha/CCL3, MIP-1 beta/CCL4, and RANTES/CCL5,
are released as chemoattractants for neutrophils, immature
dendritic cells, natural killer cells, and activated T cells
(Luster, A. D. (2002) Curr. Opin. Immunol. 14:129). Further,
several pro-inflammatory cytokines are released including IL-1
beta/IL-1F2, IL-6, and TNF-alpha/TNFSF1A. TNF-alpha also
contributes to the pro-apoptotic activity of the classically
activated macrophage (Boyle, J. J. et al. (2003) Arterioscler.
Thromb. Vasc. Biol. 23:1553; Duffield, J. S. et al. (2001) Am. J.
Pathol. 159:1397; Song, E. et al. (2000) Cell. Imunol. 204:19).
TNF-alpha is accompanied by Fas Ligand/TNFSF6 secretion and NO
release as a result of iNOS upregulation (Hesse, M. et al. (2001)
J. Immunol. 167:6533; Thomassen, M. J. & M. S. Kavuru (2001)
Int. Immunopharmacol. 1:1479; Duffield, J. S. et al. (2000) J.
Immunol 164:2110; Munder, M. et al. (1998) J. Immunol 160:5347). In
addition, the classically activated macrophage releases proteolytic
enzymes including MMP-1, -2, -7, -9, and -12, which degrade
collagen, elastin, fibronectin, and other ECM components
(Chizzolini, C. et al. (2000) J. Immunol. 164:5952; Gibbs, D. F. et
al. (1999) Am. J. Respir. Cell Mol. Biol. 20:1136; Gibbs, D. F. et
al. (1999) Am. J. Respir. Cell Mol. Biol. 20:1145).
[0221] Although the release of these molecules is important for
host defense and direction of the adaptive immune system, when
uncontrolled their release can levy significant collateral damage
on the microenvironment. By eliciting massive leukocyte
infiltration and flooding the surrounding tissue with inflammatory
mediators, pro-apoptotic factors, and matrix degrading proteases,
the classically activated macrophage is capable of dismantling
tissues to the point of inflicting serious injury. Tissue
destruction perpetrated by chronic inflammation has been associated
with the development of tumors, type 1 autoimmune diseases, and
glomerulonephritis among other pathologies (Gordon, S. (2003) Nat.
Rev. Immunol. 3:23; Mosser, D. M. (2003) J. Leukoc. Biol.
73:209).
[0222] In some embodiments, the methods of the present invention
comprise administering an oxidative compound, e.g., chlorite, or an
immunomodulating agent, e.g., an immunosuppressant, for the
treatment of a macrophage related disease including but not limited
to tumors, type 1 autoimmune diseases such as multiple sclerosis,
glomerulonephritis, rheumatoid arthritis, diabetes,
atherosclerosis, Kawasaki disease, bacteria infections, and viral
infection, e.g., HIV. In some embodiments, the present invention
provides a method for treating a macrophage related disease with an
oxidative agent or an immunomodulatory agent by modulating
IFN-gamma receptor. In some embodiments, the present invention
provides a method for treating a macrophage related disease with an
oxidative agent or an immunomodulatory agent by modulating LPS
receptor. In some embodiments, the present invention provides a
method for treating a macrophage related disease with an oxidative
agent or an immunomodulatory agent by modulating MHC II antigen
presentation pathway. In some embodiments, the present invention
provides a method for treating a macrophage related disease with an
oxidative agent or an immunomodulatory agent by modulating release
of chemokines including but not limited to IL-8/CXCL8,
IP-10/CXCL10, MIP-1 alpha/CCL3, MIP-1 beta/CCL4, and RANTES/CCL5.
In some embodiments, the present invention provides a method for
treating a macrophage related disease with an oxidative agent or an
immunomodulatory agent by modulating release of pro-inflammatory
cytokines including but not limited to IL-1 beta/IL-1F2, IL-6, and
TNF-alpha/TNFSF1A. In some embodiments, the present invention
provides a method for treating a macrophage related disease with an
oxidative agent by modulating release of proteolytic enzymes
including but not limited to MMP-1, -2, -7, -9, and -12.
[0223] II. Alternatively Activated Macrophages
[0224] Differentiation of alternatively activated macrophages does
not require any priming. IL-4 and/or IL-13 can act as sufficient
stimuli (Stein, M. et al. (1992) J. Exp. Med. 176:287; Doherty, T.
M. et al. (1993) J. Immunol. 151:7151). The binding of these
factors to their respective receptors is followed by fluid-phase
pinocytosis of soluble antigen (Brombacher, F. (2000) BioEssays
22:646; Montaner, L. J. et al. (1999) J. Immunol 162:4613; Conner,
S. D. & S. L. Schmid (2003) Nature 422:37). Soluble antigen is
then loaded onto MHC class II molecules and antigen/MHCII complexes
and co-stimulatory B7 family members are subsequently displayed to
T cells (Harding, C. V. et al. (2003) Curr. Opin. Immunol
15:112).
[0225] Similar to the classically activated macrophage, the
alternatively activated macrophage changes its cellular morphology
and secretory pattern as a result of appropriate stimulation.
Leukocytes are attracted by the macrophage via its release of
chemokines including MDC/CCL22 (Andrew, D. P. et al. (1998) J.
Immunol 161:5027; Imai, T. et al. (1999) Int. Immunol 11:81),
PARC/CCL18 (Kodelja, V. et al. (1998) J. Immunol. 160:1411; Goerdt,
S. et al. (1999) Pathobiology 67:222) and TARC/CCL17. Inflammation
is counteracted by the release of factors such as IL-1ra/IL-1F3
(Mantovani, A. et al. (2001) Trends Immunol 22:328), Ym1, Ym2,
RELMa (Raes, G. et al. (2002) J. Leukoc. Biol. 71:597; Loke, P. et
al. (2002) BMC Immunol. 3:7), IL-10, and TGF-beta. TGF-beta also
functions indirectly to promote ECM building by inducing nearby
fibroblasts to produce ECM components. The alternatively activated
macrophage itself secretes the ECM components, Fibronectin and
bIG-H3 (Gratchev, A. et al. (2001) Scand. J. Immunol 53:386), the
ECM cross-linking enzyme, Trans-glutaminase (Haroon, Z. A. et al.
(1999) Lab. Invest. 79:1679), and Osteopontin, which is involved in
cell adhesion to the ECM (Murry, C. E. et al. (1994) Am. J. Pathol.
145:1450).
[0226] In addition, alternatively activated macrophages upregulate
the enzyme Arginase I, which is involved in proline as well as
polyamine biosynthesis. Proline promotes ECM construction while
polyamines are involved in cell proliferation (Hesse, M. et al.
(2001) J. Immunol. 167:6533). Other factors secreted by the
alternatively activated macrophage that promote cell proliferation
include PDGF, IGF, and TGF-beta (Song, E. et al. (2000) Cell.
Imunol. 204:19; Cao, B. et al. (2000) Chin. Med. J. 113:776). These
factors, along with FGF basic, TGF-alpha, and VEGF, also
participate in angiogenesis (Cao, B. et al. (2000) Chin. Med. J.
113:776; Sunderkotter, C. et al. (1991) Pharmac. Ther. 51:195).
[0227] The molecules secreted by the alternatively activated
macrophage work toward resolution of inflammation and promotion of
wound repair due to their anti-inflammatory, fibrotic,
proliferative, and angiogenic activities. This macrophage is also
especially efficient at combating parasitic infections such as
Schistosomiasis. In addition to its beneficial activities, the
alternatively activated macrophage has been implicated in several
pathologies, the most prominent of which are allergy and asthma
(Duffield, J. S. (2003) Clin. Sci. 104:27; Gordon, S. (2003) Nat.
Rev. Immunol. 3:23).
[0228] In some embodiments, the methods of the present invention
comprise administering an oxidative compound, e.g., chlorite, or an
immunomodulatory agent, e.g., an immunosuppressor, for the
treatment of a macrophage related disease including but not limited
to allergy and asthma among other pathologies. In some embodiments,
the present invention provides a method for treating a macrophage
related disease with an oxidative agent or an immunomodulatory
agent by modulating the IL-4 receptor. In some embodiments, the
present invention provides a method for treating a macrophage
related disease with an oxidative agent or an immunomodulatory
agent by modulating the IL-13 receptor. In some embodiments, the
present invention provides a method for treating a macrophage
related disease with an oxidative agent or an immunomodulatory
agent by modulating release of chemokines including but not limited
to MDC/CCL22, PARC/CCL18 and TARC/CCL17. In some embodiments, the
present invention provides a method for treating a macrophage
related disease with an oxidative agent or an immunomodulatory
agent by modulating release of factors including but not limited to
IL-1ra/IL-1F3, Ym1, Ym2, RELMa, IL-10, and TGF-beta. In some
embodiments, the present invention provides a method for treating a
macrophage related disease with an oxidative agent or an
immunomodulatory agent by modulating release of ECM related
molecules including but not limited to ECM components, fibronectin
and bIG-H3, ECM cross-linking enzyme, trans-glutaminase and
osteopontin.
[0229] The present invention also encompasses methods of modulating
macrophage accumulation or activation with an oxidative agent or an
immunomodulatory agent targeting a signaling pathway including but
not limited to lipopolysaccharide (LPS), toll-like receptor (TLR),
prostaglandin E2 (PGE2), interferon (IFN)-.alpha., IFN-.beta.,
IFN-.gamma., interleukin (IL)-1, IL-4, IL-6, sIL1Ra, IL-10, IL-12,
IL-12p40, IL-13, IP10, MHC (major histocompatibility complex) Class
II molecules (MHCII), TNF-.alpha., macrophage inflammatory protein
1 alpha (MIP1-.alpha.), IFN-gamma-inducing factor (IGIF),
macrophage-stimulating protein (MSP), inter-cellular adhesion
molecule 1(ICAM-1), colony stimulating factor 1 (CSF-1R),
L-arginine, and nitric oxide signaling pathways. The oxidative or
immunomodulatory agent of the present invention may target or have
an effect on any receptor, cytosolic or nuclear intermediate
signaling molecule, or transcription factor involved in any one of
the signaling pathways disclosed herein. Examples of important
signaling molecules as part of one or more signaling pathways that
can be modulated by the oxidative agent or immunomodulatory agent
of the present invention include but are not limited to TLR2, TLR4,
CAT2, ICSBP, IL1-R, Tie-2, TRIF/IRF3, IFNR-I, IFNR-II, IRF1, IRF2,
Raf-1, MEK1, MEK2, ERK1, ERK2, p38, MAPKK4, MAPKK6, PKC, JAK1,
JAK2, STAT1, STAT3, Elk1, JNK/SAPK, AP1, Pu1, NFkB, NFAT, iNOS,
USF1, ISGF3, SP1, Bc16, ATF2, c-Jun, and COX-2. Molecules important
to macrophage activation or effects that can be modulated, either
directly or indirectly, by the oxidative agent or immunomodulatory
agent of the present invention include those that belong to
transcription factors, cell surface receptors, cytokines,
chemokines, cytokine or chemokine receptors, growth factors,
interferons, interferon receptors, and adhesion molecules.
Specifically, the oxidative or immunomodulatory agent of the
present invention can modulate molecules including but not limited
to TLR-2, TLR-4, mkp-1, COX-2, SOCS-3, Fc.gamma.R1, IFN-.alpha.,
IFN-.beta., IL-4, IL-6, IL-1Ra, IGIF, IL-.beta., MHCI, MHCII IAA,
MHCII IAB, MHCII IEB, IP10, IL-10, cathepsin H, lysozyme, CathB,
stk, TNF-.alpha., IL-12p35, IL-12p40, MIP-1a, ICAM-1, INOS, mig,
Cat-2, CIITA, ICSBP, CathL, CSF1R, GM-CSF, IRF1, IRF-2, c-fos,
VEGF, IL-8, bFGF, CSF-1, EGF, MMP-2, MMP-7, MMP-9, MMP-12, EMAPII,
endothelin 2, HIF-1, HIF-2, CXCL8, TGF.beta., PGE2, and/or MDF.
[0230] III. Monocytes
[0231] Monocytes are a type of white blood cell. Monocytes have two
main functions in the immune system: (1) replenish resident
macrophages and dendritic cells under normal states; and (2) in
response to inflammation signals, monocytes can move quickly to
sites of infection in the tissues and divide/differentiate into
macrophages and dendritic cells to elicit an immune response.
Monocytes are produced by the bone marrow from haematopoietic stem
cell precursors called monoblasts. Monocytes circulate in the
bloodstream for about one to three days and then typically move
into tissues throughout the body. In the tissues monocytes mature
into different types of macrophages at different anatomical
locations. Monocytes which migrate from the bloodstream to other
tissues will then differentiate into tissue resident macrophages or
dendritic cells. Macrophages are responsible for protecting tissues
from foreign substances but are also suspected to be the
predominant cells involved in triggering atherosclerosis. They are
cells that possess a large smooth nucleus, a large area of
cytoplasm and many internal vesicles for processing foreign
material.
[0232] There are two types of monocytes in human blood: a) the
classical monocyte, which is characterized by high level expression
of the CD14 cell surface receptor (CD14++ monocyte) and b) the
non-classical, pro-inflammatory monocyte with low level expression
of CD14 and with additional co-expression of the CD16 receptor
(CD14+CD16+ monocyte). After stimulation with microbial products
the CD14+CD16+ monocytes produce high amounts of pro-inflammatory
cytokines such as tumor necrosis factor and interleukin-12.
[0233] An increase or decrease in the number of CD14+CD16+
monocytes has been indicated in various diseases (Loems
Ziegler-Heitbrock, Journal of Leukocyte Biology, Vol 81, 2007).
These CD14+CD16+ monocytes may play a role in giving rise to
macrophages that contribute to the inflammation of a disease.
CD14+CD16+ monocytes are involved in many inflammatory diseases
including but not limited to rheumatoid arthritis, diabetes,
hemodialysis, atherosclerosis, Kawasaki disease, as well as
bacteria infections and viral infections, which are disclosed in
more details herein below. In some embodiments, the present
invention provides a method of treating a macrophage related
disease comprising administering to a subject in need thereof an
effective amount of an oxidative and/or immunomodulatory agent,
wherein the agent modulates or has an effect on CD14+CD16+
monocytes.
[0234] IV. Tumor-Associated Macrophages (TAM)
[0235] In some embodiments, the present invention provides a method
of modulating tumor associated macrophages comprising administering
an oxidative and/or an immunomodulatory agent into a subject.
Macrophages are prominent in the stromal compartment of virtually
all types of malignancy. Macrophages respond to the presence of
stimuli in different parts of tumors with the release of a distinct
repertoire of growth factors, cytokines, chemokines, and enzymes
that regulate tumor growth, angiogenesis, invasion, and/or
metastasis. The distinct microenvironments where tumor-associated
macrophages (TAM) act include: 1) areas of invasion where TAMs
promote cancer cell motility; 2) stromal and perivascular areas
where TAMs promote metastasis; and 3) avascular and perinecrotic
areas where hypoxic TAMs stimulate angiogenesis (reviewed by Lewis
C E et al. Cancer Res. 2006 (66) 605-612). TAMs have a phenotype
that are relatively immature, characterized by low expression of
the differentiation-associated macrophage antigens,
carboxypeptidase M and CD51, high constitutive expression of IL-1
and IL-6, and low expression of TNF-.alpha..
[0236] TAM infiltration correlates positively with tumor cell
proliferation as measured by MIB-1 levels in breast carcinomas,
Ki67 levels in endometrial carcinomas, or mitotic index in renal
cell carcinoma (reviewed by Lewis C E et al. Cancer Res. 2006 (66)
605-612). Various studies have shown that TAMs express a number of
factors that stimulate tumor cell proliferation and survival,
including epidermal growth factor (EGF) (Goswami S. et al. Cancer
Res 2005; 65; 5278-83; Lewis C E et al. Lancet 1993; 342; 148-9),
platelet-derived growth factor (PDGF), TGF-h1, hepatocyte growth
factor, MMP-9, and basic fibroblast growth factor (bFGF). TAMs also
play an important part in regulating angiogenesis. TAMs release a
number of potent proangiogenic cytokines and growth factors, such
as vascular endothelial growth factor (VEGF), TNF-a, IL-8, and
bFGF. Additionally, they express a broad array of
angiogenesis-modulating enzymes, including MMP-2, MMP-7, MMP-9,
MMP-12, and cyclooxygenase-2 (COX-2) (Sunderkotter C. et al.
Pharmacol Ther 1991; 51: 195-216; Klimp A H et al. Cancer Res.
2001; 61: 7305-9). TAMs respond to tumor hypoxia by upregulating
the hypoxia-inducible transcription factors HIF-1 and HIF-2.
Macrophages also upregulate VEGF and other proangiogenic factors in
response to hypoxia. For example, macrophages synthesize elevated
levels of MMP-7 when exposed to hypoxia in vitro and in avascular
areas of human tumors. A cDNA array study has identified
upregulation of messages encoding more than 30 other proangiogenic
genes in primary macrophages exposed to hypoxia, including CXCL8,
angiopoietin, COX-2 and other factors (White J R, et al. Genomics
2004; 83: 1-8).
[0237] TAMs have also been implicated in the regulation of
metastasis. High numbers of TAMs in primary tumors have been
correlated with early establishment of metastases in a number of
tumor types (Hanada T et al. Int J. Urol 2000; 7: 263-9). TAMs play
roles in both the release of metastatic cells from the primary
tumor as well as the establishment of secondary tumors at distant
sites.
[0238] TAMs also play a role in tumor immunosuppression. Unlike
macrophages from healthy tissues, which are capable of presenting
tumor-associated antigens, lysing tumor cells, and stimulating the
antitumor functions of T cells and NK cells, TAMs in the tumor
microenvironment lack these activities, leaving the host without
the ability to mount an effective antitumor immune response. A
number of studies have shown that tumor-derived molecules, like
cytokines, growth factors, chemotactic molecules, and proteases,
influence TAM functions (Elgert K D et al. J Leukoc Biol 1998; 64:
275-90). For example, tumor cells secrete proteins that can inhibit
the cytotoxic activity of TAMs, e.g., IL-4, IL-6, IL-10, MDF,
TGF-h1 and PGE 2 (Ben-Baruch, Semin Cancer Biol 2005). Moreover,
TGF-h1, IL-10, and PGE 2 may suppress the expression of MHC class
II molecules by macrophages in the tumor microenvironment as well
as distant sites like the spleen and peritoneum. This effect may
limit the ability of TAMs to present tumor-associated antigens to T
cells effectively in these areas. Another important aspect of TAM
involvement in antitumor immune mechanisms is the ability of these
cells to release immunostimulatory cytokines. For example,
macrophage expression of IL-12, a cytokine known to stimulate both
the proliferation and cytotoxicity of T cells and NK cells, is
markedly suppressed in tumors, possibly by exposure to IL-10, PGE
2, and TGF-h1 (Mitsuhashi M. et al. J Leuko Biol 2004; 76: 322-23).
Hypoxia in the tumor microenvironment is likely to contribute
suppressing the antitumor activity of TAMs as it stimulates the
release of the potent immunosuppressive factors PGE 2 and IL-10.
They act on TAMs to reduce their cytotoxicity activity toward tumor
cells. Hypoxia also inhibits the ability of macrophages to
phagocytose dead or dying cells and present antigens to T cells.
One mechanism by which this may be achieved is by reduced surface
expression of CD80, a costimulatory molecule needed for the full
activation of T-cell responses to antigenic peptides.
[0239] Many signaling pathways are important to TAM functions.
Exemplary signaling pathways regulating TAM function include but
are not limited to NFkB pathway, TLR pathways, specifically
TLR/IL-1R signaling, TLR2 and TLR4 signaling, the Tie-2/Ang-2
pathway, the TRIF/TBK1/IRF3 pathway, and hypoxia-induced pathways.
NFkB is one of the most crucial transcription factors regulating
the inflammatory repertoire of macrophages, particularly their
expression of proinflammatory cytokines, costimulatory molecules,
and other activation markers in response to diverse environmental
cues (e.g., stress signals, inflammatory cytokines, pathogens, and
hypoxia). TLR/IL-1R signaling is an important upstream component of
NFkB activation in macrophages. In inflammation-induced cancers,
activation of TLR/IL-1R on stromal macrophages may be triggered by:
1) direct interaction with bacteria at sites of chronic infection
(e.g., enteric bacteria in colitis-associated colon cancer or H.
pylori in gastric cancer) (Karin M et al. Cell 2006 124: 823-835);
or 2) interaction with tumor-cell-derived proinflammatory cytokines
like IL-1; and/or 3) recognition of components of necrotic tumor
cell debris like HMGB1 (high mobility group box 1) or S100
(reviewed by Biswas S K et al. J. Immunol. 2008 180: 2011-2017).
TLR4 activation on human lung cancer cells promotes production of
the immunosuppressive cytokine TGF-.beta. and the proangiogenic
factors VEGF and CXCL8 as well as conferring resistance to
TNF-.alpha.-induced apoptosis and tumor cell survival (He W et al.
Mol. Immunol 2007 44: 2850-2859). A preferential role of TLR2
activation in triggering an M2 (immunosuppressive)-like cytokine
profile (IL-12 low, IL-10 high) in dendritic cells and macrophages
through ERK/MAPK phosphorylation has been reported (Dillon S et al.
J Immunol 2004 172: 4733-4743).
[0240] Tie-2-expressing monocytes (TEM) exist in human and murine
tumors (De Palma et al 2005 Cancer Cell 8: 211-226). Endothelial
cells as well as tumor cells are known to up-regulate Ang-2, a
ligand for Tie-2 in tumors. It has been suggested that
tumor-derived Ang-2 may facilitate the recruitment of Tie-2
monocytes/macrophages into tumors (Murdoch C et al. J Immunol 178:
7405-7411). Importantly, Ang-2 also significantly inhibits the
release of proinflammatory cytokines like TNF-.alpha. and IL-12 by
Tie-2 monocytes in vitro (Biswas S K et al. J. Immunol. 2008 180:
2011-2017), an effect more pronounced in hypoxia. These findings
suggest that the Ang-2/Tie-2 axis may represent another potential
mechanism for dampening the antiangiogenic phenotype and prompting
the immunosuppressive phenotype of TAM, especially in hypoxic areas
of tumors.
[0241] Preferential activation of the TRIF-dependent IRF3/STAT1
pathway (where TRIF is TLR/IL-1R domain-containing adaptor inducing
IFN-.beta., TBK is TANK-binding kinase, and IRF is IFN regulatory
factor) has been demonstrated in TAM in murine fibrosarcoma (Biswas
S et al. Blood 107: 2112-2122). This was evident from the
constitutive activation of STAT1 and the up-regulation of type I
IFN-inducible genes including CCL5, CXCL9, and CXCL10 in the TAM
under basal and LPS-activated conditions (Biswas S K et al. J.
Immunol 2008 180: 2011-2017). IL-10 transcription has also been
shown to be regulated by the TRIF/IRF3 pathway via TRAF3 and type I
IFNs (Chang E Y et al. J Immunol 178: 6705-6709). Taken together,
TRIF pathway members such as TBK1 and IRF3 may play a role in
mediating the effects of TAM and may represent a potential
therapeutic target.
[0242] As mentioned hereinabove, hypoxia has profound effects on
macrophage functions including their migration into tumors and
patterns of gene expression, especially those encoding
proangiogenic cytokines and enzymes. Hypoxia induces gene
expression in these cells through up-regulation of the
transcription factors hypoxia-inducible factors (HIF) 1 and 2
(HIF-1 and HIF-2). Macrophages up-regulate both HIFs and
subsequently a wide array of HIF target genes in hypoxic/necrotic
areas of human tumors (Murdoch C et al. 2005 Int J Cancer, 117:
701-708). Most importantly, hypoxia is a potent inducer of both
VEGF and MMP7 in TAM, both of which are known to support tumor
angiogenesis, invasion, and metastasis. In addition, hypoxia
up-regulates the expression of M2 macrophage markers like IL-10,
arginase, and PGE 2. It also modulates expression of
proinflammatory genes like TNF-.alpha., IL-1, migration inhibitory
factor (MIF), CCL3, and COX2.
[0243] In some embodiments, the present invention provides a method
of treating cancer comprising administering an oxidative agent or
an immunomodulatory agent. In some embodiments, macrophage
activation or function is modulated by the oxidative or
immunomodulatory agent of the present invention such that the
antitumor activity is enhanced. In some embodiments, the oxidative
agent of the present invention modulates one or more pathways
involved in macrophage activation or function, wherein the pathways
include but are not limited to the NFkB pathway, TLR pathway,
Tie-2/Ang-2 pathway, TRIF/TBK1/IRF3 pathway, hypoxia-induced
pathway and any pathway involving any molecule disclosed
herein.
D. Methods of Treatment of Disease
[0244] The present invention provides methods for the treatment of
a variety of diseases and disorders using an oxidative agent,
including but not limited to chlorite or chlorite-containing
agents, alone or in combination with another agent effective in
treating the disease. The present invention also provides methods
for the treatment of a variety of diseases and disorders using an
immunomodulatory agent, including but not limited to
immunosuppressive agents, alone or in combination with another
agent effective in treating the disease.
[0245] In some embodiments, an oxidative agent and an
immunomodulatory agent are used together for combination therapy.
By combination therapy is meant the simultaneous administration of
an immunomodulator and an oxidative agent in the same formulation
or in separate formulations, or the administration of an
immunomodulator and an oxidative agent at separate times. In one
aspect, the present invention provides for a method of treating a
disease that responds to an immunomodulator by administering to a
subject in need thereof an immunomodulator in combination with an
oxidative agent. In an alternative embodiment, the present
invention provides for a method of ameliorating the side effects of
immunomodulator therapy, the method comprising administering an
oxidative agent to a subject experiencing or at risk of
experiencing side effects from immunomodulator therapy.
[0246] Unless the context indicates otherwise, all of the
formulations and pharmaceutical formulations described herein may
be used in the methods of treatment provided by the invention. As
used herein and as well understood in the art, examples of
treatment include obtaining beneficial or desired results,
including clinical results. Non-limiting examples of beneficial or
desired clinical results include one or more of alleviation or
amelioration of one or more symptoms, diminishment of extent of a
condition, including a disease, stabilized (i.e., not worsening)
state of a condition, including diseases, preventing spread of
disease, delay or slowing of condition, including disease,
progression, amelioration or palliation of the condition, including
disease, state, and remission (whether partial or total), whether
detectable or undetectable. In some variations, oxidative and/or
immunomodulatory agents as described herein are used to achieve one
or more of treating, preventing, delaying the onset of, or causing
the regression of the diseases or conditions described herein.
[0247] Reference herein to "treatment" and "prophylaxis" is to be
considered in its broadest context. The term "treatment" does not
necessarily imply that a subject is treated until total recovery.
Similarly, "prophylaxis" does not necessarily mean that the subject
will not eventually contract a disease condition. Accordingly,
treatment and prophylaxis can include amelioration of the symptoms
of a particular condition or preventing or otherwise reducing the
risk of developing a particular condition. The term "prophylaxis"
may be considered as reducing the severity or onset of a particular
condition. "Treatment" may also reduce the severity of an existing
condition. Treating or treatment of a disease or condition with
oxidative and/or immunomodulatory agents includes: (1) preventing
or reducing the risk of developing the disease, i.e., causing the
clinical symptoms of the disease not to develop in a subject that
may be exposed to or predisposed to the disease but does not yet
experience or display symptoms of the disease; (2) inhibiting the
disease, i.e., arresting or reducing the development of the disease
or its clinical symptoms; or (3) relieving the disease, i.e.,
causing regression of the disease or its clinical symptoms. In some
embodiments, treatment may be understood to not include prevention.
The term "modulate" can refer to the treating, prevention,
suppression, enhancement or induction of a function or condition.
For example, compounds can modulate macrophage activation by
enhancing macrophage activation or inhibiting macrophage
activation. In some embodiments, the term "modulate" may be
understood to not include prevention of a function or
condition.
[0248] A "subject" or "patient" includes humans and non-human
mammals. Examples of subject or patients include, but are not
limited to mice, rats, hamsters, guinea pigs, pigs, rabbits, cats,
dogs, goats, sheep, cows, and humans. The term "mammal" includes,
without limitation, humans, domestic animals (e.g., dogs or cats),
farm animals (cows, horses, or pigs), and laboratory animals (mice,
rats, hamsters, guinea pigs, pigs, rabbits, dogs, or monkeys).
[0249] The terms "disease" and "disorder" are used interchangeably
herein. In general, a therapeutically effective amount of a
formulation is administered to a subject. An "effective amount,"
which is also referred to herein as a "therapeutically effective
amount," of a therapeutic agent for administration as described
herein is that amount of the therapeutic agent that provides the
therapeutic effect sought when administered to the subject. A
therapeutically effective amount may be achieved in a single
administration or after multiple administrations. The achieving of
different therapeutic effects may require different effective
amounts of therapeutic agent. For example, the therapeutically
effective amount of a therapeutic agent used for preventing a
disease or condition may be different from the therapeutically
effective amount used for treating, inhibiting, delaying the onset
of, or causing the regression of the disease or condition. In
addition, the therapeutically effective amount may depend on the
age, weight, the bioavailability of the compound, the severity of
the disease or condition, and other health conditions of the
subject as is well know to those versed in the disease or condition
being addressed. Thus, the therapeutically effective amount may not
be the same in every subject to which the therapeutic agent is
administered. In some embodiments, the therapeutically effective
amount of an oxidative or immunomodulatory agent of the invention
is reduced by combination therapy with another oxidative or
immunomodulatory agent, or other therapy or intervention.
[0250] To determine whether a level of therapeutic agent is a
"therapeutically effective amount" to treat the diseases or
conditions described herein, the chlorite formulations may be
administered in appropriate animal models for the diseases or
conditions of interest, and the effects may be observed to
determine whether the treatment was effective in the animal model.
The appropriate level for a different subject, including but not
limited to a human subject, may be estimated using methods known by
those of skill in the art.
[0251] Effective dosages may be estimated initially from in vitro
assays. For example, an initial dosage for use in animals may be
formulated to achieve a circulating blood or serum concentration of
active compound that is at or above an IC50 of the particular
compound as measured in an in vitro assay. Calculating dosages to
achieve such circulating blood or serum concentrations, taking into
account the bioavailability of the particular active agent, is well
within the capabilities of skilled artisans. For guidance, the
reader is referred to Fingl & Woodbury, "General Principles,"
In: Goodman and Gilman's The Pharmaceutical Basis of Therapeutics,
Chapter 1, pp. 1-46, latest edition, Pergamagon Press, which is
hereby incorporated by reference in its entirety, and the
references cited therein.
[0252] The oxidative agent or immunomodulatory agent of the present
invention can be administered alone or as part of a combination
therapy, e.g., administered in combination with or adjunctive to
other therapies or interventions for treating the diseases or
conditions described herein. Administration of the oxidative agent
or immunomodulatory agent may be prior to, subsequent to, or
concurrent with one or more other treatments, including but not
limited to treatments using other active agents or
non-pharmaceutical therapies such as radiotherapy. In some
variations the oxidative agent, e.g., chlorite, or immunomodulatory
agent are used in accordance with their standard or common dosages,
as specified in the prescribing information accompanying other
commercially available chlorite formulations. See, e.g., the
prescribing information in the 2005 Edition of The Physician's Desk
Reference, the disclosures of which are incorporated herein by
reference.
[0253] Combination therapy includes administration of a single
pharmaceutical dosage formulation that contains an oxidative agent,
e.g., chlorite as described herein, and/or an immunomodulatory
agent, e.g., an immunosuppressive agent, and one or more additional
active agents, as well as administration of an oxidative agent and
each active agent in its own separate pharmaceutical dosage
formulation. In a non-limiting example, chlorite and a DPP-IV
inhibitor can be administered to a subject having Type II diabetes
together in a single dosage composition, or each agent can be
administered in separate dosage formulations. Where separate dosage
formulations are used, oxidative agents, immunomodulatory agents,
and/or one or more additional active agents can be administered at
essentially the same time (i.e., concurrently), or at separately
staggered times (i.e., sequentially). Combination therapy is
understood to include all these regimens.
[0254] One of skill in the art will appreciate that the effective
amount may be adjusted when therapeutic agents are used in
combination with the oxidative or immunomodulatory agents of the
present invention. In a non-limiting example, a chlorite
composition provided by the present invention may be used in
combination with a DPP4 inhibitor, e.g., sitagliptin. When such
combinations are used, the dose of one or more of the agents may be
reduced to a level below the level required for a desired efficacy
when the one or more agents are used alone.
[0255] The term "diagnosing" refers to determining the presence or
absence of a particular disease or condition. Additionally, the
term refers to determining the level or severity of a particular
disease or condition, as well as monitoring of the disease or
condition to determine its response to a particular therapeutic
regimen.
[0256] In one aspect, the present invention provides a method of
treating a macrophage related disease comprising administering to a
subject in need thereof an effective amount of a pharmaceutical
composition comprising an oxidative agent, e.g., chlorite or a
chlorite-containing agent, or an immunomodulatory agent, e.g., an
immunosuppressant. In some embodiments, the chlorite composition
further comprises a pH adjusting agent and a pharmaceutically
acceptable excipient. In another aspect, the present invention
provides a method of treating a macrophage related disease
comprising administering to a subject in need thereof an effective
amount of an oxidative and/or immunomodulatory agent. In some
embodiments, an oxidative agent and an immunomodulatory agent are
used together for combination therapy. In the present invention,
the term "macrophage related diseases" encompasses diseases related
to monocytes as well.
[0257] Due to their role in phagocytosis, macrophages are involved
in many diseases of the immune system. For example, they
participate in the formation of granulomas, which are inflammatory
lesions that are caused by a large number of diseases. Some
disorders of ineffective phagocytosis and macrophage function have
been described. Macrophages are the predominant cells involved in
creating the progressive plaque lesions of atherosclerosis (Lucas A
D, Greaves D R Expert Rev Mol Med 3 (25): 1-18). Macrophages also
play a role in human immunodeficiency virus (HIV) infection.
Similar to T cells, macrophages can be infected with HIV, and
become a reservoir of ongoing virus replication throughout the
body. Furthermore, macrophages are believed to help cancer cells
proliferate as well. They are attracted to oxygen-starved (hypoxic)
tumor cells and promote chronic inflammation. Inflammatory
compounds such as tumor necrosis factor (TNF) released by the
macrophage activates the gene switch nuclear factor-kappa B. NF-kB
then enters the nucleus of a tumor cell and turns on production of
anti-apoptotic proteins that prevent apoptosis and promote cell
proliferation and inflammation (Gary Stix 2007 Scientific American:
46-49). Several non-limiting examples of macrophage related
diseases are described below.
[0258] Diseases related to macrophage presence or function are
encompassed within the scope of the methods of the present
invention. In one embodiment, macrophage related diseases are
diseases characterized by activated macrophages. The macrophages
may be chronically activated or acutely activated, or both. While
not wishing to be bound by theory, treatments according to certain
embodiments of the invention may interfere with the activation of
monocytes to macrophages, or may increase deactivation of
macrophages, or both.
[0259] Macrophage related diseases that can be treated or prevented
by the methods of the present invention include but are not limited
to cancer, autoimmune diseases such as multiple sclerosis and
rheumatoid arthritis, macrophage activation syndrome,
atherosclerosis, diabetes mellitus, Kawasaki disease, asthma,
hemophagocytic lymphohistiocytosis, sarcoidosis, periodontitis,
Whipple's disease, pulmonary alveolar proteinosis, macrophage
related pulmonary disease, Leishmaniasis, obesity complications,
hemodialysis related inflammation, microbial infection, retroviral
infection such as HIV infection, and inflammation. In addition, for
many of the diseases, although macrophages may not be the primary
trigger of the disease, their involvement is evident in the disease
related complications or secondary manifestations. Such macrophage
related complications can be treated or prevented by the methods of
the present invention. For example, macrophages contribute to
neurological signs in acquired immunodeficiency syndrome (AIDS) and
non-AIDS-related diseases. Other examples of complications that may
involve macrophages and can be treated with the subject methods
include but are not limited to transplant-related complications,
acute atheroma complication, metabolic syndrome, hypertension,
obesity, diabetic complications (nephropathy, neuropathy and
retinopathy), complications of the tobacco-related disease, liver
complications, inflammatory neurological diseases, and a variety of
other disorders. In one embodiment, the diseases to be treated
exclude neurodegenerative diseases. In one embodiment, the diseases
to be treated are limited to diseases of the organs of the thorax.
Alternatively, the diseases to be treated involve the peripheral
nervous system.
[0260] In one embodiment, the diseases to be treated according to
the invention are related to or can be identified by elevated
levels of soluble CD14 (sCD14) in plasma. Such diseases may include
hemodialysis patients, periodontitis, chronic heart failure,
pulmonary tuberculosis, oral lichen planus and burning mouth
syndrome, inflammatory bowel disease, carrageenan-primed endotoxin
shock mouse model, sepsis, inactive Crohn's Disease, brucellosis,
severe acute pancreatitis, atopic syndrome including atopic
conditions in children, aortic stiffness, chronic Hepatitis B and
C, transplant-related conditions including lung transplant
patients, multiple organ dysfunction syndrome, Non-Hodgkin's
lymphoma, systemic lupus erythematosus, status asthmaticus, Lyme
Disease, arthritis such as rheumatoid arthritis, Kawasaki Disease,
acute respiratory distress syndrome, scleroderma, acute plasmodium
falciparum malaria, and sarcoidoisis. In one embodiment, the
diseases to be treated according to the invention are related to or
can be identified by elevated levels of soluble CD14 (sCD14) in
plasma which can be attributed to a pathogen. Such diseases may
include periodontitis, pulmonary tuberculosis, oral lichen planus
and burning mouth syndrome, carrageenan-primed endotoxin shock
mouse model, sepsis, brucellosis, chronic Hepatitis B and C, Lyme
Disease, pathogen-induced Kawasaki Disease, and acute plasmodium
falciparum malaria. In one embodiment, the diseases to be treated
according to the invention are related to or can be identified by
elevated levels of soluble CD14 (sCD14) in plasma where the
diseases do not directly involve the central nervous system.
Alternately, the disease to be treated may have an unknown
etiology, such as arthritics, asthma, scleroderma, inflammatory
bowel disease, systemic lupus erythematosus, and atopic
syndrome.
[0261] In one embodiment, the diseases to be treated according to
the invention are related to or can be identified by elevated
levels of soluble CD163 (sCD163) in plasma. Such diseases may
include inflammatory disorders and sepsis, Type 2 diabetes, acute
malaria, bacteraemia, acute liver failure, fulminant hepatic
failure, multiple sclerosis, Macrophage Activation Syndrome,
pneumoniae bacteremia, coeliac disease, Hepatitis C and B, coronary
atherosclerosis, reactive hemophagocytic syndrome, Gaucher's
Disease, myeloid leukaemia, and rheumatoid arthritis. In one
embodiment, the diseases to be treated according to the invention
are related to or can be identified by elevated levels of soluble
CD163 (sCD163) in plasma which can be attributed to a pathogen.
Such diseases may include sepsis, acute malaria, bacteraemia,
pneumoniae bacteremia, and Hepatitis C and B, Alternately, the
disease to be treated may have an unknown etiology, or a
non-pathogenic etiology.
[0262] In one embodiment, the diseases to be treated according to
the invention are related to or can be identified by elevated
levels of more than one biomarker in plasma. For example, levels of
sCD14 and sCD163 may both be measured and correlated. In certain
embodiments, treatment according to the invention reduces the
elevated levels of biomarker, such as sCD14 and/or sCD163.
[0263] In one preferred embodiment, the disease to be treated shows
upregulation of differentiation of monocytes to activated
macrophages, where treatment according to the invention reduces
such upregulation. The reduction in upregulation may in some
embodiments occur before upregulation, or alternatively, by
downregulation after the fact, or both. In one embodiment,
macrophage deactivation is characterized by levels of expression of
CD16 in monocytes or macrophages, for example, median CD16
expression levels in CD14+ cells. A preferred oxidative agent
according to this embodiment is an N-halo compound such as
1,3-dichloro-5,5-dimethylhydantoin or chloramine T, alone or in
combination with an immunomodulatory agent such as natalizumab
(Tysabri.RTM.), fingolimod, or cladribine. In one embodiment, the
disease to be treated is multiple sclerosis or Crohn's disease.
[0264] Diseases to be treated according to the invention include
but are not limited to the following diseases.
[0265] I. Metabolic and Vascular Disorders
[0266] In some embodiments, the methods of the present invention
comprise administration of oxidative agent and/or immunomodulatory
agent in combination with other therapeutic agents and/or
interventions that are commonly used for the treatment of diabetes
and related disorders. In such embodiments, combination therapy can
be used for modulating (preventing the onset of the symptoms or
complications associated with) diabetes, or treating, preventing or
reducing the risk of developing, diabetes and its related symptoms,
complications, and disorders. In non-limiting examples, an
oxidative agent and/or immunomodulatory agent can be used in
combination with, for example, biguanides (such as metformin);
thiazolidinediones (such as ciglitazone, pioglitazone,
troglitazone, and rosiglitazone); dipeptidyl-peptidase-4 ("DPP-IV")
inhibitors (such as vildagliptin, sitagliptin, saxagliptin,
linagliptin and alogliptin); glucagonlike peptide-1 ("GLP-1")
receptor agonists (such as exanatide) or GLP-1 mimetics; PPAR gamma
agonists or partial agonists; dual PPAR alpha, PPAR gamma agonists
or partial agonists; dual PPAR delta, PPAR gamma agonists or
partial agonists; pan PPAR agonists or partial agonists;
dehydroepiandrosterone (also referred to as DHEA or its conjugated
sulphate ester, DHEA-SO.sub.4); antiglucocorticoids; TNF-alpha
inhibitors; alpha-glucosidase inhibitors (such as acarbose,
miglitol, and voglibose); sulfonylureas (such as chlorpropamide,
tolbutamide, acetohexamide, tolazamide, glyburide, gliclazide,
glynase, glimepiride, and glipizide); pramlintide (a synthetic
analog of the human hormone amylin); other insulin secretogogues
(such as repaglinide, gliquidone, and nateglinide); insulin (or
insulin mimetics); glucagon receptor antagonists; gastric
inhibitory peptide ("GIP"); or GIP mimetics.
[0267] In some embodiments, the methods of the present invention
comprise administration of an oxidative agent and/or
immunomodulatory agent in combination with other therapeutic agents
and/or interventions that are commonly used for the treatment of
complications of macrophage related diseases. Such agents include
but are not limited to agents used to treat vascular disease,
including but not limited to endothelin receptor antagonists
commonly used for the treatment of hypertension and other
endothelial dysfunction-related disorders, such as bosentan,
darusentan, enrasentan, tezosentan, atrasentan, ambrisentan
sitaxsentan; smooth muscle relaxants such as PDE5 inhibitors
(indirect-acting) and minoxidil (direct-acting); angiotensin
converting enzyme (ACE) inhibitors such as captopril, enalapril,
lisinopril, fosinopril, perindopril, quinapril, trandolapril,
benazepril, ramipril; angiotensin II receptor blockers such as
irbesartan, losartan, valsartan, eprosartan, olmesartan,
candesartan, telmisartan; beta blockers such as atenolol,
metoprolol, nadolol, bisoprolol, pindolol, acebutolol, betaxolol,
propranolol; diuretics such as thiazide, hydrochlorothiazide,
furosemide, torsemide, metolazone; calcium channel blockers such as
amlodipine, felodipine, nisoldipine, nifedipine, verapamil,
diltiazem; alpha receptor blockers doxazosin, terazosin, alfuzosin,
tamsulosin; and central alpha agonists such as clonidine. Such
agents include but are not limited to agents used to treat
hyperlipidemia, including but not limited to agents that lower LDL
such as statins (atovastatin, fluvastatin, lovastatin, pravastatin,
rosuvastatin calcium, simvastatin) and nicotinic acid, agents that
stimulate PPAR alpha such as fibrates, gemfibrozil, fenofibrate,
bezafibrate, ciprofibrate, agents that bind and prevent
readsorption of bile acids and reduce cholesterol levels such as
bile acid sequestrants, cholestyramine and colestipol, and
cholesterol absorption inhibitors. Such agents include those that
reduce risk of heart attack, including COX-1 inhibitors including
aspirin and non-steroidal anti-inflammatory drugs (NSAIDs), or
COX-2 inhibitors. Such agents include but are not limited to agents
used to treat microbial infections, including antibiotics,
antiviral and antifungal. Such agents include but are not limited
to agents used to treat inflammation, including various
immunosuppressants, for example glucocorticoids, non-glucocorticoid
steroids, cytostatics, antibodies, and drugs acting on
immunophilins. Examples of immunosuppressants that can be used in
combination with the oxidative agents of the present invention for
treatment of autoimmune diseases and various inflammatory diseases
include but are not limited to a cytostatic, such as an alkylating
agent, an antimetabolite, or a cytotoxic antibiotic, a folic acid
analog such as methotrexate, or a purine analog such as
azathioprine and mercaptopurine, an antibody, a TNF binding
protein, methotrexate, azathioprine, mercaptopurine, an interferon,
an opioid, or mycophenolate, a calcineurin inhibitor such as
cyclosporin, tacrolimus, sirolimus, or an analog of any
thereof.
[0268] In some embodiments, the methods of the present invention
comprise administration of an oxidative agent and/or
immunomodulatory agent in combination with other therapeutic agents
and/or interventions that are used for the treatment of obesity or
obesity-related disorders. In some embodiments, the methods
comprise administration of the oxidative and/or immunomodulatory
agent in combination with, e.g., phenylpropanolamine, phenteramine;
diethylpropion; mazindol; fenfluramine; dexfenfluramine;
phentiramine, beta-3 adrenoceptor agonist agents; sibutramine;
gastrointestinal lipase inhibitors (such as orlistat); and leptins.
Other agents used in treating obesity or obesity-related disorders
wherein the compounds of the present invention can be effectively
used in combination with, e.g., cannabinoid-1 ("CB-1") receptor
antagonists (such as rimonabant); PPAR delta agonists or partial
agonists; dual PPAR alpha, PPAR delta agonists or partial agonists;
dual PPAR delta, PPAR gamma agonists or partial agonists; pan PPAR
agonists or partial agonists; neuropeptide Y; enterostatin;
cholecytokinin; bombesin; amylin; histamine H3 receptors; dopamine
D2 receptors; melanocyte stimulating hormone; corticotrophin
releasing factor; galanin; and gamma amino butyric acid (GABA).
[0269] In some embodiments, the methods of the present invention
comprise administration of an oxidative agent and/or
immunomodulatory agent in combination with other therapeutic agents
and/or interventions that are used for the treatment of
hyperlipidemia and related complications, e.g., statins (such as
atorvastatin, fluvastatin, lovastatin, pravastatin, and
simvastatin), CETP inhibitors (such as torcetrapib); a cholesterol
absorption inhibitor (such as ezetimibe); PPAR alpha agonists or
partial agonists; PPAR delta agonists or partial agonists; dual
PPAR alpha, PPAR delta agonists or partial agonists; dual PPAR
alpha, PPAR gamma agonists or partial agonists; dual PPAR delta,
PPAR gamma agonists or partial agonists; pan PPAR agonists or
partial agonists; fenofibric acid derivatives (such as gemfibrozil,
clofibrate, fenofibrate, and bezafibrate); bile acid-binding resins
(such as colestipol or cholestyramine); nicotinic acid; probucol;
betacarotene; vitamin E; or vitamin C.
[0270] In some embodiments, the methods of the present invention
comprise administration of an oxidative agent and/or
immunomodulatory agent in combination with other therapeutic agents
and/or interventions that are used for the treatment of
atherosclerosis. In some embodiments, the methods comprise
administration of the oxidative and/or immunomodulatory agent in
combination with one or more of the following active agents: an
antihyperlipidemic agent; a plasma HDL-raising agent; an
antihypercholesterolemic agent, such as a cholesterol biosynthesis
inhibitor, e.g., an hydroxymethylglutaryl (HMG) CoA reductase
inhibitor (also referred to as statins, such as lovastatin,
simvastatin, pravastatin, fluvastatin, and atorvastatin); an
HMG-CoA synthase inhibitor; a squalene epoxidase inhibitor; or a
squalene synthetase inhibitor (also known as squalene synthase
inhibitor); an acyl-coenzyme A cholesterol acyltransferase (ACAT)
inhibitor, such as melinamide; probucol; nicotinic acid and the
salts thereof and niacinamide; a cholesterol absorption inhibitor,
such as beta-sitosterol; a bile acid sequestrant anion exchange
resin, such as cholestyramine, colestipol or dialkylaminoalkyl
derivatives of a cross-linked dextran; an LDL receptor inducer;
fibrates, such as clofibrate, bezafibrate, fenofibrate, and
gemfibrizol; vitamin B6 (also known as pyridoxine) and the
pharmaceutically acceptable salts thereof, such as the HCl salt;
vitamin B12 (also known as cyanocobalamin); vitamin B3 (also known
as nicotinic acid and niacinamide); anti-oxidant vitamins, such as
vitamin C and E and beta carotene; a beta-blocker; an angiotensin
II antagonist; an angiotensin converting enzyme inhibitor; PPAR
alpha agonists or partial agonists; PPAR delta agonists or partial
agonists; PPAR gamma agonists or partial agonists; dual PPAR alpha,
PPAR delta agonists or partial agonists; dual PPAR alpha, PPAR
gamma agonists or partial agonists; dual PPAR delta, PPAR gamma
agonists or partial agonists; pan PPAR agonists or partial
agonists; and a platelet aggregation inhibitor, such as fibrinogen
receptor antagonists (i.e., glycoprotein IIb/IIIa fibrinogen
receptor antagonists) and aspirin. The oxidative agent, e.g.,
chlorite, or immunomodulatory agent, eg, immunosuppressor, can be
administered in combination with more than one additional active
agent, e.g., a combination of chlorite with an HMG-CoA reductase
inhibitor (e.g., atorvastatin, fluvastatin, lovastatin,
pravastatin, and simvastatin) and aspirin, or in combination with
an HMG-CoA reductase inhibitor and a blocker.
[0271] In some embodiments, the methods of the present invention
comprise administration of an oxidative agent and/or
immunomodulatory agent in combination with other therapeutic agents
and/or interventions that are used for the treatment of
hyperlipidemia. In some embodiments, the methods comprise
administration of the oxidative and/or immunomodulatory agent
combination with one or more of the following active agents for
modulating hyperlipidemia (treating hyperlipidemia and its related
complications) including but not limited to statins (such as
atorvastatin, fluvastatin, lovastatin, pravastatin, and
simvastatin), CETP inhibitors (such as torcetrapib); a cholesterol
absorption inhibitor (such as ezetimibe); PPAR alpha agonists or
partial agonists; PPAR delta agonists or partial agonists; dual
PPAR alpha, PPAR delta agonists or partial agonists; dual PPAR
alpha, PPAR gamma agonists or partial agonists; dual PPAR delta,
PPAR gamma agonists or partial agonists; pan PPAR agonists or
partial agonists; fenofibric acid derivatives (such as gemfibrozil,
clofibrate, fenofibrate, and bezafibrate); bile acid-binding resins
(such as colestipol or cholestyramine); nicotinic acid; probucol;
betacarotene; vitamin E; or vitamin C.
[0272] Additionally, the methods of the present invention comprise
administration of an oxidative agent and/or immunomodulatory agent
in combination with a therapeutically effective amount of one or
more active agents selected from the group consisting of: an
antihyperlipidemic agent; a plasma HDL-raising agent; an
antihypercholesterolemic agent, such as a cholesterol biosynthesis
inhibitor, for example, an HMG-CoA reductase inhibitor; an HMG-CoA
synthase inhibitor; a squalene epoxidase inhibitor, or a squalene
synthetase inhibitor (also known as squalene synthase inhibitor);
an acyl-coenzyme A cholesterol acyltransferase inhibitor; probucol;
nicotinic acid and the salts thereof; CETP inhibitors such as
torcetrapib; a cholesterol absorption inhibitor such as ezetimibe;
PPAR alpha agonists or partial agonists; PPAR delta agonists or
partial agonists; dual PPAR alpha, PPAR delta agonists or partial
agonists; dual PPAR alpha, PPAR gamma agonists or partial agonists;
dual PPAR delta, PPAR gamma agonists or partial agonists; pan PPAR
agonists or partial agonists; niacinamide; a cholesterol absorption
inhibitor; a bile acid sequestrant anion exchange resin; a LDL
receptor inducer; clofibrate, fenofibrate, and gemfibrozil; vitamin
B6 and the pharmaceutically acceptable salts thereof; vitamin B 12;
an anti-oxidant vitamin; a beta-blocker; an angiotensin II
antagonist; an angiotensin converting enzyme inhibitor; a platelet
aggregation inhibitor; a fibrinogen receptor antagonist; aspirin;
phentiramines, beta-3 adrenergic receptor agonists; sulfonylureas,
biguanides, alpha-glucosidase inhibitors, other insulin
secretogogues, and insulin.
[0273] In some embodiments, the present invention provides a method
to administer an oxidative agent and/or immunomodulatory agent in
combination with one or more other therapeutic agents and/or
interventions used for the treatment of metabolic syndrome (or
treating metabolic syndrome and its related symptoms, complications
and disorders), wherein the compounds of the present invention can
be effectively used in combination with, for example, the active
agents discussed above for modulating or treating diabetes,
obesity, hyperlipidemia, atherosclerosis, and/or their respective
related symptoms, complications and disorders.
[0274] In a further embodiment, the present invention provides a
method to administer an oxidative agent and/or immunomodulatory
agent in combination with halofenic acid, an ester of halofenic
acid, or another prodrug of halofenic acid, preferably with
(-)-(4-chlorophenyl)-(3-trifluoromethylphenoxy)-acetic acid
2-acetylaminoethyl ester (metaglidasen).
[0275] In some embodiments, the present invention provides a method
that comprises administering an oxidative agent and/or
immunomodulatory agent in combination with one or more allergy
drugs for the treatment of diabetes and obesity. Two common
over-the-counter allergy medications are effective in reducing
complications of both obesity and Type 2 diabetes in mice. The
medications, ketotifen fumarate and cromolyn, stabilize a
population of inflammatory immune cells called mast cells in people
suffering from asthma or allergy. Theses anti-allergy or asthma
drugs may have therapeutic effects for the treatment of macrophage
related diseases including but not limited to diabetes and obesity
in humans. Researchers have also found that a regulatory T cell
also acts as a liaison between the metabolic and immune systems-in
this case, controlling inflammation in fat tissue. Fat tissue from
obese and insulin-resistant mice and people is marked by a dramatic
absence of regulatory T cells or Tregs, in dramatic contrast to an
already reported overabundance in fat tissue of inflammatory
macrophages (Calisha, "Common allergy drug reduces obesity and
diabetes in mice", Fierce Biotech July 2009). Although obese and
diabetic fat tissue was full of inflammatory macrophages and nearly
absent of Tregs, normal-weight fat tissue was the diametric
opposite. It is possible that the inflammation caused by
macrophages results in insulin resistance. It is further likely
that Tregs keep the macrophages in check in normal fat tissue, thus
preventing inflammation.
[0276] II. Macrophage-Activation Syndrome
[0277] In some embodiments, the present invention provides a method
of treating a macrophage-activation syndrome comprising
administering an effective amount of an oxidative agent, including
but not limited to chlorite, chlorite containing agents, or
derivatives thereof. In some embodiments, the present invention
provides a method of treating a macrophage-activation syndrome
comprising administering an effective amount of an immunomodulatory
agent, including but not limited to immunosuppressants. In some
embodiments, an oxidative agent and an immunomodulatory agent are
used together for combination therapy to treat
macrophage-activation syndrome.
[0278] Macrophage-activation syndrome (MAS) is a severe,
potentially life-threatening, complication of several chronic
rheumatic diseases of childhood. It occurs most commonly with
systemic-onset juvenile idiopathic arthritis (SoJIA), which is also
known as Still's disease. In addition, MAS has been described in
association with diseases including but not limited to systemic
lupus erythematosus (SLE), Kawasaki disease, and adult-onset
Still's disease. It is thought to be closely related and
pathophysiologically very similar to reactive (secondary)
hemophagocytic lymphohistiocytosis (HLH). The hallmark clinical and
laboratory features include high fever, hepatosplenomegaly,
lymphadenopathy, pancytopenia, liver dysfunction, disseminated
intravascular coagulation, hypofibrinogenemia, hyperferritinemia,
and hypertriglyceridemia. Despite marked systemic inflammation, the
erythrocyte sedimentation rate (ESR) is paradoxically depressed,
caused by low fibrinogen levels. A bone marrow biopsy or aspirate
usually shows hemophagocytosis. There is uncontrolled activation
and proliferation of macrophages and T lymphocytes, with a marked
increase in circulating cytokines, such as IFN-gamma, and
granulocyte-macrophage colony-stimulating factor (GM-CSF). In many
cases of MAS, decreased natural killer cell (NK-cell) function is
observed. Most commonly used treatments include high-dose
glucocorticoids, and cyclosporine. In refractory cases treatment
regimens are used similar to that in HLH (Pinto L, et al. J Assoc
Physicians India. 2007 55:185-7).
[0279] III. Autoimmune Diseases
[0280] In some embodiments, the present invention provides a method
of treating a macrophage related disease comprising administering
an effective amount of an oxidative agent, including but not
limited to chlorite, chlorite containing agents, or derivatives
thereof, wherein the macrophage related disease is an autoimmune
disease. In some embodiments, the present invention provides a
method of treating a macrophage related disease comprising
administering an effective amount of an immunomodulatory agent,
including but not limited to immunosuppressants, wherein the
macrophage related disease is an autoimmune disease. In some
embodiments, an oxidative agent and an immunomodulatory agent are
used together for combination therapy to treat autoimmune
diseases.
[0281] Autoimmunity is the failure of an organism to recognize its
own constituent parts as self, which allows an immune response
against its own cells and tissues. Diseases that results from such
aberrant immune response can be considered autoimmune diseases.
Prominent examples include Coeliac disease, diabetes mellitus type
1 (IDDM), systemic lupus erythematosus (SLE), Sjogren's syndrome,
Churg-Strauss Syndrome, Hashimoto's thyroiditis, Graves' disease,
idiopathic thrombocytopenic purpura, and rheumatoid arthritis (RA).
The treatment of autoimmune diseases is typically
immunosuppressive, anti-inflammatory, or palliative. Hormone levels
have been shown to affect the severity of some autoimmune diseases
such as multiple sclerosis. Other causes may include the presence
of fetal cells in the maternal bloodstream, i.e., microchimerism,
and infections with some viruses and bacteria. The autoimmune
diseases that can be treated with the methods of the present
invention include but are not limited to acute disseminated
encephalomyelitis, Addison's disease, alopecia areata, ankylosing
spondylitis, antiphospholipid antibody syndrome, autoimmune
hemolytic anemia, autoimmune hepatitis, Bullous pemphigoid, Coeliac
disease, Crohns disease, dermatomyositis, diabetes mellitus type 1,
Goodpasture's syndrome, Graves' disease, Guillain-Barre syndrome,
Hashimoto's disease, Idiopathic thrombocytopenic purpura, Lupus
erythematosus, Mixed Connective Tissue Disease, multiple sclerosis,
myasthenia gravis, narcolepsy, Pemphigus vulgaris, Pernicious
anemia, Polymyositis, Primary biliary cirrhosis, Rheumatoid
arthritis, Sjogren's syndrome, Temporal arteritis, Ulcerative
Colitis, Vasculitis, and Wegener's granulomatosis.
[0282] IV. Multiple Sclerosis
[0283] In some embodiments, the present invention provides a method
of treating multiple sclerosis comprising administering an
effective amount of an oxidative agent, including but not limited
to chlorite, chlorite containing agents, or derivatives thereof. In
some embodiments, the present invention provides a method of
treating multiple sclerosis comprising administering an effective
amount of an immunomodulatory agent, including but not limited to
immunosuppressants. In some embodiments, an oxidative agent and an
immunomodulatory agent are used together for combination therapy to
treat multiple sclerosis.
[0284] Multiple sclerosis (MS) is an autoimmune disease in which a
body's immune response attacks the central nervous system, leading
to demyelination (Compston A, Coles A 2002 Lancet 359
(9313):1221-31). MS affects the ability of nerve cells in the brain
and spinal cord to communicate with each other. Nerve cells
communicate by sending electrical signals called action potentials
down axons, which are wrapped in myelin. In MS, the body's own
immune system destroys oligodendrocytes, the cells responsible for
creating and maintaining the myelin sheath, which helps the neurons
carry electrical signals. MS results in a thinning or complete loss
of myelin, and, as the disease advances, the cutting (transection)
of the neuron's extensions or axons. When the myelin is lost, a
neuron can no longer effectively conduct electrical signals. The
name multiple sclerosis refers to scars in the white matter of the
brain and spinal cord, which is mainly composed of myelin. Almost
any neurological symptom can appear with the disease, and often
progresses to physical and cognitive disability and
neuropsychiatric disorder.
[0285] Apart from demyelination, inflammation is the other
pathologic hallmark of MS. According to a strictly immunological
explanation of MS, the inflammatory process is caused by T cells.
In MS, T cells gain entry into the brain via the blood-brain
barrier. The blood-brain barrier is normally not permeable to these
types of cells, unless triggered by infection or a virus, which
decreases the integrity of the tight junctions forming the barrier.
When the blood-brain barrier regains its integrity, usually after
infection or virus has cleared, the T cells are trapped inside the
brain. The T cells recognize myelin as foreign and attack it as if
it were an invading virus. This triggers inflammatory processes,
stimulating other immune cells and the production of soluble
factors including cytokines and antibodies. Leaks form in the
blood-brain barrier, which in turn cause a number of other damaging
effects such as swelling, activation of macrophages, and more
activation of cytokines and other destructive proteins. Macrophages
are present within active plaques of MS patients (L. Steinman, A
molecular trio in relapse and remission in multiple sclerosis,
Nature Reviews Immunology 9, 440-447 (2009)). Immunohistochemistry
with macrophage markers shows that in the pathogenesis of
experimental autoimmune encephalomyetitis, which is a mouse disease
model of multiple sclerosis, different populations of macrophages
(i.e., perivascular cells, microglia and infiltrating blood-borne
macrophages) are present in the central nervous system (Bauer J, et
al. The Histochemical Journal 28:83-97, 1996).
[0286] Administration of high doses of intravenous corticosteroids,
such as methylprednisolone, has been the standard therapy for acute
relapses of MS. Several studies have shown that treatment with
interferons during an initial attack can decrease the chance that a
patient will develop MS. These results support the use of
interferon after a first clinical demyelinating event and indicate
that there may be modest beneficial effects of immediate treatment
compared with delayed initiation of treatment (Jacobs L D, et al.
(2000). N Engl J Med 343 (13): 898-904). Several disease-modifying
treatments have been approved by regulatory agencies of different
countries. Interferon formulations including those of interferon
beta-1a (Avonex.RTM., which is injected weekly; Rebif.RTM.,
injected three times a week; and CinnoVex.TM., a
biosimilar/biogeneric), and interferon beta-1b (Betaseron.RTM. in
the U.S. and Betaferon.RTM. in Europe and Japan, injected every
other day). Another MS medication is glatiramer acetate
(Copaxone.RTM.), injected daily, a mixture of polypeptides which
may protect myelin proteins by substituting itself as the target of
immune system attack (Ziemssen T, Schrempf W (2007). Int. Rev.
Neurobiol. 79: 537-70). Still another approved MS medication,
mitoxantrone, is an immunosuppressant also used in cancer
chemotherapy. Finally, natalizumab is a humanized monoclonal
antibody against the cellular adhesion molecule .alpha.4-integrin
and is believed to reduce the ability of inflammatory immune cells
to attach to and pass through the cell layers lining the intestines
and blood-brain barrier.
[0287] In some embodiments, the present invention provides a method
of treating multiple sclerosis by modulating macrophage activation
with an oxidative agent, including but not limited to chlorite and
chlorite-containing agents. In some embodiments, the present
invention provides a method of treating multiple sclerosis by
modulating macrophage activation with an immunomodulatory agent. In
some embodiments, the macrophage activation is reduced or
inhibited.
[0288] V. Rheumatoid Arthritis
[0289] In some embodiments, the present invention provides a method
of treating rheumatoid arthritis comprising administering an
effective amount of an oxidative agent including but not limited to
chlorite, a chlorite containing agent, or a derivative or any
thereof. In some embodiments, the present invention provides a
method of treating rheumatoid arthritis comprising administering an
effective amount of an immunomodulatory agent, e.g., an
immunosuppressant. In some embodiments, an oxidative agent and an
immunomodulatory agent are used together for combination therapy to
treat rheumatoid arthritis.
[0290] Rheumatoid arthritis (RA) is a chronic, systemic
inflammatory disorder that may affect many tissues and organs, but
principally attacks the joints producing an inflammatory synovitis
that often progresses to destruction of the articular cartilage and
ankylosis of the joints. Rheumatoid arthritis can also produce
diffuse inflammation in the lungs, pericardium, pleura, and sclera,
and also nodular lesions, most common in subcutaneous tissue under
the skin. Although the cause of rheumatoid arthritis is unknown,
autoimmunity plays a pivotal role in its chronicity and
progression. The rheumatoid nodule, which is often subcutaneous, is
the feature most characteristic of rheumatoid arthritis. The nodule
has a central area of fibrinoid necrosis that may be fissured and
which corresponds to the fibrin-rich necrotic material found in and
around an affected synovial space. Surrounding the necrosis is a
layer of palisading macrophages and fibroblasts, corresponding to
the intimal layer in synovium and a cuff of connective tissue
containing clusters of lymphocytes and plasma cells, corresponding
to the subintimal zone in synovitis.
[0291] RA is an autoimmune disease. Once the abnormal immune
response has become established, plasma cells derived from B
lymphocytes produce rheumatoid factors and ACPA of the IgG and IgM
classes in large quantities. These are not deposited in the way
that they are in systemic lupus. Rather, they appear to activate
macrophages through Fc receptor and perhaps complement binding This
can contribute to inflammation of the synovium, in terms of edema,
vasodilation and infiltration by activated T-cells (mainly CD4 in
nodular aggregates and CD8 in diffuse infiltrates). Synovial
macrophages and dendritic cells further function as antigen
presenting cells by expressing MHC class II molecules, leading to
an established local immune reaction in the tissue. The disease
progresses in concert with formation of granulation tissue at the
edges of the synovial lining (pannus) with extensive angiogenesis
and production of enzymes that cause tissue damage. Modern
pharmacological treatments of RA target these mediators. Once the
inflammatory reaction is established, the synovium thickens, the
cartilage and the underlying bone begins to disintegrate and
evidence of joint destruction accrues.
[0292] Pharmacological treatment of RA can be divided into
disease-modifying antirheumatic drugs (DMARDs), anti-inflammatory
agents and analgesics (Vital E, Emery P (2005) Am Fam Physician 72
(6): 1002, 1004). Traditional small molecular mass drugs include
chemically synthesized DMARDs such as azathioprine, ciclosporin
(cyclosporine A), D-penicillamine, gold salts, hydroxychloroquine,
leflunomide, methotrexate (MTX), minocycline, and sulfasalazine
(SSZ). Biological agents (biologics) are produced through genetic
engineering, and include but are not limited to tumor necrosis
factor alpha (TNF.alpha.) blockers--etanercept (Enbrel.RTM.),
infliximab (Remicade.RTM.), adalimumab (Humira.RTM.); Interleukin 1
(IL-1) blockers--anakinra (Kineret.RTM.); monoclonal antibodies
against B cells--rituximab (Rituxan.RTM.); T cell costimulation
blocker--abatacept (Orencia.RTM.); and Interleukin 6 (IL-6)
blockers--tocilizumab (an anti-IL-6 receptor antibody) (RoActemra,
Actemra.TM.). Anti-inflammatory agents include but are not limited
to glucocorticoids and non-steroidal anti-inflammatory drug
(NSAIDs). Analgesics include but are not limited to paracetamol
(acetaminophen in US and Canada); opiates, diproqualone, and
lidocaine topical. Other therapies for RA include but are not
limited to weight loss, occupational therapy, podiatry,
physiotherapy, immunoadsorption therapy, joint injections, and
special tools to improve hard movements.
[0293] The abundance and activation of macrophages in the inflamed
synovial membrane/pannus significantly correlates with the severity
of rheumatoid arthritis (RA). Although unlikely to be the
`initiators` of RA, macrophages possess widespread
pro-inflammatory, destructive, and remodeling capabilities that can
critically contribute to acute and chronic disease. Also,
activation of the monocytic lineage is not locally restricted, but
extends to systemic parts of the mononuclear phagocyte system
(Kinne R W et al, Arthritis Res. 2000; 2(3):189-202). A rise in the
presence of CD14+ CD16+ monocytes has been shown in active RA
(Kawanaka N, et al, Arthritis Rheum 46, 2578-2586). Macrophages in
synovial fluid also express CD16 and so do macrophages in the
synovial membrane-lining layer (Wahl S M, J Immunol 1992, 148,
485-490; Iwahashi M, et al, Arthritis Rheum 2004, 50, 1457-1467).
Thus, selective counteraction of macrophage activation remains an
efficacious approach to diminish local and systemic inflammation,
as well as to prevent irreversible joint damage.
[0294] In some embodiments, the present invention provides a method
of treating rheumatoid arthritis by modulating macrophage
activation with an oxidative agent, including but not limited to
chlorite and chlorite-containing agents. In some embodiments, the
present invention provides a method of treating rheumatoid
arthritis by modulating macrophage activation with an
immunomodulatory agent, including but not limited to
immunosuppressants. In some embodiments, the macrophage activation
is reduced or inhibited.
[0295] VI. Atherosclerosis
[0296] In some embodiments, the present invention provides a method
of treating atherosclerosis comprising administering an effective
amount of an oxidative agent, including but not limited to
chlorite, chlorite-containing agents or derivatives thereof. In
some embodiments, the present invention provides a method of
treating atherosclerosis comprising administering an effective
amount of an immunomodulatory agent, including but not limited to
immunosuppressants. In some embodiments, an oxidative agent and an
immunomodulatory agent are used together for combination therapy to
treat atherosclerosis.
[0297] Atherosclerosis is the condition in which an artery wall
thickens as the result of a build-up of fatty materials such as
cholesterol. It is a syndrome affecting arterial blood vessels, a
chronic inflammatory response in the walls of arteries, in large
part due to the accumulation of macrophage white blood cells and
promoted by low density (especially small particle) lipoproteins
(plasma proteins that carry cholesterol and triglycerides) without
adequate removal of fats and cholesterol from the macrophages by
functional high density lipoproteins (HDL). It is caused by the
formation of multiple plaques within the arteries (Maton, et al.
(1993). Human Biology and Health. Englewood Cliffs, N.J., USA:
Prentice Hall). The atheromatous plaque is divided into three
distinct components: the atheroma, which is the nodular
accumulation at the center of large plaques, composed of
macrophages nearest the lumen of the artery; underlying areas of
cholesterol crystals; and calcification at the outer base of
older/more advanced lesions.
[0298] The first step of atherogenesis is the development of fatty
streaks, which are small sub-endothelial deposits of
monocyte-derived macrophages. The primary documented driver of this
process is oxidized lipoprotein particles within the wall, beneath
the endothelial cells, though upper normal or elevated
concentrations of blood glucose also plays a major role and not all
factors are fully understood. Fatty streaks may appear and
disappear. Low Density Lipoprotein particles in blood plasma, when
they invade the endothelium and become oxidized, create a risk for
cardiovascular disease. A complex set of biochemical reactions
regulates the oxidation of LDL, chiefly stimulated by presence of
enzymes, e.g., Lp-LpA2 and free radicals in the endothelium or
blood vessel lining
[0299] The initial damage to the blood vessel wall results in an
inflammatory response. Monocytes enter the artery wall from the
bloodstream, with platelets adhering to the area of insult. This
may be promoted by redox signaling induction of factors such as
VCAM-1, which recruit circulating monocytes. The monocytes
differentiate macrophages, which ingest oxidized LDL, slowly
turning into large "foam cells"--so-described because of their
changed appearance resulting from the numerous internal cytoplasmic
vesicles and resulting high lipid content. Foam cells eventually
die, and further propagate the inflammatory process. There is also
smooth muscle proliferation and migration from tunica media to
intima responding to cytokines secreted by damaged endothelial
cells. This would cause the formation of a fibrous capsule covering
the fatty streak.
[0300] In terms of treatment for atherosclerosis, in general, the
group of medications referred to as statins has been the most
popular and are widely prescribed for treating atherosclerosis. The
statins, and some other medications, have been shown to have
antioxidant effects, possibly part of their basis for some of their
therapeutic success in reducing cardiac events. Combinations of
statins, niacin, intestinal cholesterol absorption-inhibiting
supplements (ezetimibe and others, and to a much lesser extent
fibrates) have been the most successful in changing common but
sub-optimal lipoprotein patterns and group outcomes. Diet and
dietary supplements are also used to help treat atherosclerosis.
For example, vitamin C acts as an antioxidant in vessels and
inhibits inflammatory process (Bohm F, et al. (2007)
Atherosclerosis 190 (2): 408-15). Patients at risk for
atherosclerosis-related diseases are increasingly being treated
prophylactically with low-dose aspirin and a statin.
[0301] The actions of macrophages drive atherosclerotic plaque
progression Immunomodulation of atherosclerosis is the term for
techniques which modulate immune system function in order to
suppress this macrophage action (Jan Nilsson; et al (2005)
Arteriosclerosis, Thrombosis, and Vascular Biology 5: 18-28). In
some embodiments, the present invention provides a method of
treating atherosclerosis by modulating macrophage accumulation or
activation with an oxidative agent, including but not limited to
chlorite and chlorite-containing agents. In some embodiments, the
present invention provides a method of treating atherosclerosis by
modulating macrophage accumulation or activation with an
immunomodulatory agent, including but not limited to
immunosuppressants. In some embodiments, the macrophage activation
is reduced or inhibited.
[0302] VII. Stroke and Spinal Cord Pathology
[0303] In some embodiments, the present invention provides methods
for treatment of stroke, post-stroke brain damage or spinal cord
pathology with an oxidative agent, including but not limited to
chlorite and chlorite-containing agents. In some embodiments, the
present invention provides methods for treatment of stroke,
post-stroke brain damage or spinal cord pathology with an
immunomodulatory agent, including but not limited to
immunosuppressants. In some embodiments, an oxidative agent and an
immunomodulatory agent are used together for combination therapy to
treat stroke, post-stroke brain damage or spinal cord
pathology.
[0304] A stroke is the rapidly developing loss of brain function(s)
due to disturbance in the blood supply to the brain due to ischemia
(lack of blood supply) caused by thrombosis, embolism or
hemorrhage. Certain classes of immune cells flood the brain soon
after a stroke, causing inflammation and more neurological damage.
Mice deficient in these immune cells may suffer far less brain
damage after a stroke compared to normal mice. While the initial
damage from a stroke cannot be prevented, drugs can be used to
limit secondary damage caused by immune cells that rush to the site
of the infarction, or stroke. The initial damage happens
immediately after a stroke, it cannot be efficiently blocked
because it is very rapid. But after this neural damage, macrophages
and T-cells are recruited and this inflammation induces the growth
of the infarction. This secondary damage can be blocked by
suppressing the inflammation. In mice with induced stroke, it has
been observed that the subsequent recruitment of immune cells
caused inflammation and more damage. One of the first cytokines
found expressed within the brain was interleukin-23 (IL-23). IL-23
activates other immune cells including T-cells and macrophages,
which then attack the brain. Mice genetically engineered to be
deficient in IL-23 suffered the least brain damage. IL-23 operates
immediately after stroke or one day later. Thus the sooner the
intervention to block IL-23, the more protective it is for the
brain.
[0305] Most patients suffering from stroke come to hospital within
a day after a stroke. A therapeutic method could prevent further
the expansion of infarction by modulating macrophages and their
infiltration to the inflammatory or injury site.
[0306] In some embodiments, the present invention provides methods
for treatment of stroke with an oxidative agent, including but not
limited to chlorite and chlorite-containing agents. In some
embodiments, the present invention provides methods for treatment
of stroke with an immunomodulatory agent, including but not limited
to immunosuppressants. In some embodiments, the treatment is
administered in combination with blockers of IL-23, e.g.,
ustekinumab, an anti-IL-23 antibody. In some embodiments, the
oxidative or immunosuppressive agent of the present invention can
be used to treat other inflammatory diseases, for example,
inflammatory bowel disease.
[0307] VIII. Diabetes Mellitus
[0308] In some embodiments, the present invention provides methods
for treatment of Diabetes mellitus (diabetes) with an oxidative
agent, including but not limited to chlorite and
chlorite-containing agents. For example, in one aspect, the present
invention provides a method of treating Type II diabetes comprising
administering to a subject in need thereof an effective amount of a
pharmaceutical composition comprising: a) chlorite, b) a pH
adjusting agent, and c) a pharmaceutically acceptable excipient. In
some embodiments, the present invention provides a method of
treating diabetic complications comprising administering to a
subject in need thereof an effective amount of a pharmaceutical
composition comprising: a) chlorite, b) a pH adjusting agent, and
c) a pharmaceutically acceptable excipient. In some embodiments,
the present invention provides methods for treatment of Diabetes
mellitus (diabetes) and/or diabetic complications with an
immunosuppressive agent, including but not limited to
immunosuppressants. In some embodiments, an oxidative agent and an
immunomodulatory agent are used together for combination therapy to
treat Diabetes mellitus.
[0309] Diabetes mellitus, or diabetes, refers to a disease or
condition that is generally characterized by metabolic defects in
production and utilization of glucose that result in the failure to
maintain appropriate blood sugar levels in the body. The result of
these defects is elevated blood glucose, referred to as
"hyperglycemia." Other types of disorders of glucose homeostasis
include impaired glucose tolerance, which is a metabolic stage
intermediate between normal glucose homeostasis and diabetes, and
gestational diabetes mellitus, which is glucose intolerance in
pregnancy in women without previous diabetic history.
[0310] Diabetes is a metabolic syndrome, which refers to a cluster
of metabolic abnormalities including obesity, insulin resistance,
glucose intolerance, diabetes, hypertension and dyslipidemia. These
abnormalities are known to be associated with an increased risk of
vascular events.
[0311] Diabetes can be divided into two clinical syndromes, Type I
and Type II diabetes mellitus. According to the American Diabetes
Association, about 8% of the U.S. population has diabetes and the
numbers are rising. See
www.diabetes.org/diabetes-statistics/prevalence.jsp. About 90% of
these cases are Type II diabetes.
[0312] Type I diabetes, or insulin-dependent diabetes mellitus, is
a chronic autoimmune disease characterized by the extensive loss of
beta cells in the pancreatic islets of Langerhans, which produce
insulin. As these cells are progressively destroyed, the amount of
secreted insulin decreases, eventually leading to hyperglycemia
(abnormally high level of glucose in the blood) when the amount
secreted drops below the level required for euglycemia (normal
blood glucose level).
[0313] Type II diabetes, or non-insulin-dependent diabetes
mellitus, develops when muscle, fat and liver cells fail to respond
normally to insulin. This failure to respond (called insulin
resistance) may be due to reduced numbers of insulin receptors on
these cells, or a dysfunction of signaling pathways within the
cells, or both. The beta cells initially compensate for this
insulin resistance by increasing their insulin output. Over time,
these cells become unable to produce enough insulin to maintain
normal glucose levels, indicating progression to Type II
diabetes.
[0314] The fasting hyperglycemia that characterizes Type II
diabetes occurs as a consequence of the combined lesions of insulin
resistance and beta cell dysfunction. The beta cell defect has two
components: the first component, an elevation of basal insulin
release (occurring in the presence of low, non-stimulatory glucose
concentrations), is observed in obese, insulin-resistant
pre-diabetic stages as well as in Type II diabetes. The second
component is a failure to increase insulin release above the
already elevated basal output in response to a hyperglycemic
challenge. This lesion is absent in pre-diabetes and appears to
define the transition from normo-glycemic insulin-resistant states
to frank diabetes.
[0315] Insulin resistance includes the diminished ability of
insulin to exert its biological action across a broad range of
concentrations producing less than the expected biologic effect
(see, e.g., Reaven G M, J. Basic & Clin. Phys. & Pharm.
9:387-406 and Flie J, Ann Rev. Med. (1983) 34:145-60). Insulin
resistant persons have a diminished ability to properly metabolize
glucose and respond poorly, if at all, to insulin therapy.
Manifestations of insulin resistance include insufficient insulin
activation of glucose uptake, oxidation and storage in muscle and
inadequate insulin repression of lipolysis in adipose tissue and of
glucose production and secretion in liver. Insulin resistance can
cause or contribute to polycystic ovarian syndrome, impaired
glucose tolerance, gestational diabetes, metabolic syndrome,
hypertension, obesity, atherosclerosis and a variety of other
disorders. Eventually, the insulin resistant individuals can
progress to a point where a diabetic state is reached.
[0316] Symptoms of diabetes, include, but are not limited to,
polyuria, polydipsia, and polyphagia. "Polyuria" refers to the
passage of a large volume of urine during a given period;
"polydipsia" refers to chronic, excessive thirst; and "polyphagia"
refers to excessive eating. Other symptoms of diabetes include,
e.g., increased susceptibility to certain infections (especially
fungal and staphylococcal infections), nausea, and ketoacidosis
(enhanced production of ketone bodies in the blood).
[0317] Diabetic complications include, but are not limited to,
microvascular complications and macrovascular complications.
Microvascular complications are those complications that generally
result in small blood vessel damage. These complications include,
e.g., retinopathy (the impairment or loss of vision due to blood
vessel damage in the eyes); neuropathy (nerve damage and foot
problems due to blood vessel damage to the nervous system);
gastroparesis (impaired ability of the stomach to empty contents
due to nerve damage, e.g., to the vagus nerve); skin complications
(bacterial infections, fungal infections, itching, dermopathy,
necrobiosis lipoidica diabeticorum, diabetic blisters, and eruptive
xanthomatosis); and nephropathy (kidney disease due to blood vessel
damage in the kidneys) and other renal disorders. Macrovascular
complications are those complications that generally result from
large blood vessel damage. These complications include, e.g.,
cardiovascular disease and peripheral vascular disease.
[0318] Cardiovascular disease refers to diseases of blood vessels
of the heart. See, e.g., Kaplan R M, et al., "Cardiovascular
diseases" in Health and Human Behavior, pp. 206-242 (McGraw-Hill,
New York 1993). Cardiovascular disease is generally one of several
forms, including, e.g., hypertension (also referred to as high
blood pressure), coronary heart disease, stroke, and rheumatic
heart disease. Peripheral vascular disease refers to diseases of
any of the blood vessels outside of the heart. It is often a
narrowing of the blood vessels that carry blood to leg and arm
muscles. Cardiovascular disorders include, but are not limited to,
hypertension, myocardial infraction, metabolic syndrome, ischemic
cardiac disease, coronary artery disease, cerebrovascular disease,
vascular dementia, preeclampsia, heart disease, stroke,
atherogenesis, thrombogenesis, atheroscleorsis, inflammatory
disease or peripheral, carotid, or coronary vascular disease.
[0319] Other conditions related to diabetes include metabolic
syndrome, hyperlipidemia, obesity, and adverse effects thereof.
[0320] The guidelines for diagnosis of Type II diabetes and related
conditions, e.g., impaired glucose tolerance and gestational
diabetes, have been outlined by the American Diabetes Association
(see, e.g., The Expert Committee on the Diagnosis and
Classification of Diabetes Mellitus, Diabetes Care, (1999) Vol 2
(Suppl 1):55-19).
[0321] There is currently no cure for Type II diabetes.
Conventional treatments are limited, and focus on attempting to
control blood glucose levels in order to minimize or delay
complications. Type II diabetics can suffer from both hypoglycemia
and hyperglycemia. Hypoglycemia is also called insulin reaction.
Signs of hypoglycemia include shakiness, dizziness, sweating,
hunger, headache, pale skin, sudden moodiness or behavior changes,
clumsiness or jerkiness, difficulty paying attention, tingling in
the mouth, or in some cases seizure. Immediate care comprises sugar
intake, e.g., in the form of sugar pills, candy, or direct
injection of glucagon. Signs and symptoms of hyperglycemia include
high blood glucose, high levels of sugar in the urine, frequent
urination, and increased thirst. Failure to treat hyperglycemia can
lead to ketoacidosis, or diabetic coma. Ketoacidosis occurs when
the body does not have enough insulin and to use glucose as fuel,
and instead breaks down fat to provide energy. The latter process
produces ketones as a by-product, which builds up in the body and
can result in coma or death.
[0322] Diabetes control involves diligent glucose monitoring and
some Type II diabetics require insulin. Insulin is broken down
during digestion and is administered through injections, insulin
pumps, through the skin, or inhaled. Several other classes of
therapeutic agents are also available. Conventional oral
medications sold today in the United States belong to one of five
classes of drugs: sulfonylureas, meglitinides, biguanides,
thiazolidinediones, DPP4 inhibitors and alpha-glucosidase
inhibitors. These treatments can act as sensitizers to reduce
insulin resistance, e.g., metformin or thiazolidinediones ("TZDs"),
or can act as secretagogues to enhance insulin secretion from beta
cells, e.g., sulphonylureas or meglitinides.
[0323] Sulphonylureas stimulate beta cells to produce more insulin.
They have the side effect of hypoglycemia since they cause insulin
secretion independent of circulating glucose levels. First
generation sulphoylureas include chlorpropamide, which is still in
use, and tolbutamide, acetohexamide and tolazamide. The second
generation sulfonylureas are used in smaller doses than the
first-generation drugs. These include glipizide, glyburide,
gliclazide and glimepiride.
[0324] Meglitinides, e.g., repaglinide and nateglinide, also
stimulate beta cells to release insulin. They can be taken before
meals to boost insulin response to food. Like sulfonylureas, they
can have the side effect of hypoglycemia. They can also lead to
weight gain.
[0325] Biguanides include the first-line treatment metformin.
Biguanides lower blood glucose levels primarily by reducing the
amount of glucose produced by the liver. They may also make muscle
tissue more amenable to glucose uptake. Side effects of metformin
include diarrhea. Other biguanides include phenformin and buformin,
both of which were withdrawn over lactic acidosis risk.
[0326] Similar to biguanides, thiazolidinediones help insulin work
better in the muscle and fat and also reduce glucose production in
the liver. Thiazolidinediones activate PPARs (peroxisome
proliferator-activated receptors), specifically PPARy, leading to a
number of effects including lowering insulin resistance. Approved
members include troglitazone, rosiglitazone and pioglitazone,
although troglitazone was pulled from the market for causing
drug-induced hepatitis. Other TZDs include MCC-555, rivoglitazone,
and ciglitazon. In some patients, TZDs cause water retention which
may lead to edema and heart failure. A related class of drugs with
potential to treat symptoms of metabolic disease includes "dual,"
"balanced" or "pan" PPAR agonists. These include aleglitazar,
muraglitazar and tesaglitazar.
[0327] Sitagliptin, sold by Merck under the trade name Januvia.TM.,
is the first approved drug of a class of compounds that inhibit
dipeptidyl peptidase IV (DPP4). These drugs can increase blood
levels of incretin hormones, which can further increase insulin
secretion, reduce glucagon secretion and have other less well
characterized effects. However, sitagliptin and other DPP4
inhibitors may also influence the tissue levels of other hormones
and peptides, and the long-term consequences of this broader effect
have not been fully investigated. In addition to sitagliptin, a
number of DPP4 inhibitors have been developed, includin
vildagliptin, saxagliptin, linagliptin and alogliptin.
[0328] Alpha-glucosidase inhibitors include acarbose, meglitol and
voglibose. These drugs do not directly effect insulin secretion of
sensitivity. Rather, they inhibit the breakdown of starches and
some sugars, thereby helping to maintain lower blood glucose levels
after a meal. They are taken with a meal. Side effects include gas,
diarrhea and weight gain.
[0329] Two other injectable drugs have been approved for Type II
diabetes. The first, pramlintide, is a synthetic form of the
hormone amylin. Amylin is secreted from beta cells along with
insulin, and interacts with insulin to help maintain blood glucose
levels. The second approved drug, exanatide, is an agonist of the
incretin glucagon-like peptide-1 (GLP-1). Incretins are a group of
hormones that increase insulin secretion in response to food
intake. Incretins include GLP-1 and gastric inhibitory peptide
("GIP"). GLP-1 stimulates insulin secretion but is rapidly degraded
by DPP4. Exanatide stimulates insulin secretion only in the
presence of high glucose, but is not orally available and must be
injected. Similar drugs in development include liraglutide, a GLP-1
analog, albiglutide, and taspoglatide.
[0330] The above drugs are often taken in combination. For example,
metformin and a sulfonylurea may be used together.
[0331] Despite the currently available treatments, the incidence of
diabetes is rising and has reached epidemic proportions. There is a
continued need for improved treatments. In some embodiments, the
present invention provides methods for treatment of diabetes using
an oxidative agent, including but not limited to chlorite and
chlorite-containing agents. In some embodiments, the present
invention provides methods for treatment of diabetes using an
immunomodulatory agent, including but not limited to
immunosuppressants.
[0332] IX. Kawasaki Disease
[0333] In some embodiments, the present invention provides a method
of treating Kawasaki disease comprising administering an effective
amount of an oxidative agent, including but not limited to chlorite
and chlorite-containing agents. In some embodiments, the present
invention provides a method of treating Kawasaki disease comprising
administering an effective amount of an immunomodulatory agent,
including but not limited to immunosuppressive agent. In some
embodiments, an oxidative agent and an immunomodulatory agent are
used together for combination therapy to treat Kawasaki
disease.
[0334] Kawasaki disease (also known as lymph node syndrome,
mucocutaneous node disease, infantile polyarteritis and Kawasaki
syndrome) is an inflammation (vasculitis) of the middle-sized
arteries. It affects many organs, including the skin, mucous
membranes, lymph nodes, and blood vessel walls, but the most
serious effect is on the heart where it can cause severe aneurysmal
dilations. There is often a pre-existing viral infection that may
play some role in pathogenesis. The conjunctival and oral mucosa,
along with the epidermis (skin), become erythmatous (red and
inflammed). Edema is often seen in the hands and feet and the
cervical lymph nodes are often enlarged.
[0335] Like other autoimmune diseases, the cause of Kawasaki
disease is presumably the interaction of genetic and environmental
factors, possibly including an infection. The specific cause is
unknown, but current theories center primarily on immunological
causes for the disease. An association has been identified with a
SNP in the ITPKC gene, which codes an enzyme that negatively
regulates T-cell activation (Onouchi Y, Gunji T, Burns J C, et al.
2008 Nat. Genet. 40 (1): 35-42). The HLA-B51 serotype has been
found to be associated with endemic instances of the disease.
[0336] Intravenous immunoglobulin (IVIG) is the standard treatment
for Kawasaki disease (Oates-Whitehead R M, et al. (2003) Cochrane
Database Syst Rev (4)) and is administered in high doses with
marked improvement usually noted within 24 hours. Salicylate
therapy, particularly aspirin, remains an important part of the
treatment but salicylates alone are not as effective as Intravenous
immunoglobulin. Corticosteroids have also been used, especially
when other treatments fail or symptoms recur.
[0337] In the acute phase of the Kawasaki disease, an expansion of
the CD14+CD16+ monocytes has been described (Nakatani K, et al.,
1999 Clin. Exp. Immunol, 117, 418-422). In some embodiments, the
present invention provides a method of treating Kawasaki disease by
modulating macrophage accumulation or activation with an oxidative
agent, including but not limited to chlorite and
chlorite-containing agents. In some embodiments, the present
invention provides a method of treating Kawasaki disease by
modulating macrophage accumulation or activation with an
immunomodulatory agent, including but not limited to
immunosuppressive agent. In some embodiments, the macrophage
activation is reduced or inhibited.
[0338] X. Asthma
[0339] In some embodiments, the present invention provides a method
of treating asthma comprising administering an effective amount of
an oxidative agent, including but not limited to chlorite and
chlorite-containing agents. In some embodiments, the present
invention provides a method of treating asthma comprising
administering an effective amount of an immunomodulatory agent,
including but not limited to immunosuppressive agent. In some
embodiments, an oxidative agent and an immunomodulatory agent are
used together for combination therapy to treat asthma.
[0340] Asthma is a chronic inflammation of the lungs in which the
airways (bronchi) are reversibly narrowed. During attacks
(exacerbations), the smooth muscle cells in the bronchi constrict,
and the airways become inflamed and swollen. Attacks can be
prevented by avoiding triggering factors and by drug treatment.
Drugs are used for acute attacks, commonly inhaled
.beta.2-agonists. In more serious cases, drugs are used for
long-term prevention, starting with inhaled corticosteroids, and
then long-acting .beta.2-agonists if necessary. Leukotriene
antagonists are less effective than corticosteroids but have no
side effects. Monoclonal antibodies such as mepolizumab and
omalizumab are sometimes effective. Prognosis is good with
treatment. Symptomatic control of episodes of wheezing and
shortness of breath is generally achieved with fast-acting
bronchodilators. Relievers include but are not limited to
short-acting, selective beta.sub.2-adrenoceptor agonists, such as
salbutamol, levalbuterol, terbutaline and bitolterol. Older, less
selective adrenergic agonists, such as inhaled epinephrine and
ephedrine tablets, have also been used. Anticholinergic
medications, such as ipratropium bromide may be used instead.
Inhaled glucocorticoids are usually considered preventive
medications while oral glucocorticoids are often used to supplement
treatment of a severe attack. Long-acting bronchodilators (LABD)
are similar in structure to short-acting selective
beta.sub.2-adrenoceptor agonists, but have much longer side chains
resulting in a 12-hour effect, and are used to give a smoothed
symptomatic relief. Currently available long-acting
beta.sub.2-adrenoceptor agonists include salmeterol, formoterol,
bambuterol, and sustained-release oral albuterol. Combinations of
inhaled steroids and long-acting bronchodilators are becoming more
widespread; the most common combination currently in use is
fluticasone/salmeterol. Another combination is
budesonide/formoterol which is commercially known as Symbicort.
[0341] Expansion of the CD14+ CD16+ monocytes has been shown in
asthma (Rivier A et al, Clin. Exp. Immunol, 100, 314-318). The
alveolar macrophage is the predominant immune effector cell
resident in the alveolar spaces and conducting airways, and it is
responsible for activating inflammatory responses sufficient to
eliminate the interlopers (Hocking, W., and D. Golde. 1979. N.
Engl. J. Med. 301:580-587, 639-644). Alveolar macrophage may exert
anti-asthmatic effects (see American Journal of Respiratory Cell
and Molecular Biology. Vol. 31, pp. 3-7, 2004).
[0342] In some embodiments, the present invention provides a method
of treating asthma by modulating macrophage accumulation or
activation with an oxidative agent, including but not limited to
chlorite and chlorite-containing agents. In some embodiments, the
present invention provides a method of treating asthma by
modulating macrophage accumulation or activation with an
immunomodulatory agent. In one example, the macrophage is alveolar
macrophage. In some embodiments, the alveolar macrophages are
activated for the treatment of asthma.
[0343] XI. Hemophagocytic Lymphohistiocytosis
[0344] In some embodiments, the present invention provides a method
of treating hemophagocytic lymphohistiocytosis comprising
administering an effective amount of an oxidative agent, including
but not limited to chlorite and chlorite-containing agents. In some
embodiments, the present invention provides a method of treating
hemophagocytic lymphohistiocytosis comprising administering an
effective amount of an immunomodulatory agent, including but not
limited to immunosuppressive agents. In some embodiments, an
oxidative agent and an immunomodulatory agent are used together for
combination therapy to treat hemophagocytic
lymphohistiocytosis.
[0345] Hemophagocytic lymphohistiocytosis, abbreviated HLH and also
known as hemophagocytic syndrome, is an uncommon hematologic
disorder that, typically, clinically manifests as fever,
splenomegaly and jaundice, with laboratory findings of
lymphocytosis and histiocytosis, and has the pathologic finding of
hemophagocytosis. HLH can arise in a number of settings and is
thought to arise from T-cell dysregulation. HLH comprises familial
(primary) hemophagocytic lymphohistiocytosis (FHL) and secondary
HLH (SHLH). FHL, an autosomal recessive disorder, is associated
with defective triggering of apoptosis and reduced cytotoxic
activity, resulting in a widespread accumulation of T lymphocytes
and activated macrophages. HLH also shows expansions of CD14+ CD16+
monocytes (Emminger W et al, Eur J. Immunol. 31, 1716-1719). In
terms of treatment, etoposide, cyclosporin, and methotrexate are
among the medications that have been proposed (Henter J I, et al.
2002 Blood 100 (7): 2367-2373). Use of the CHOP protocol has been
described. 1003231h some embodiments, the present invention
provides a method of treating hemophagocytic lymphohistiocytosis by
modulating macrophage accumulation or activation with an oxidative
agent, including but not limited to chlorite and
chlorite-containing agents. In some embodiments, the present
invention provides a method of treating hemophagocytic
lymphohistiocytosis by modulating macrophage accumulation or
activation with an immunomodulatory agent, including but not
limited to immunosuppressive agents. In some embodiments, the
macrophage activation is reduced or inhibited.
[0346] XII. Sarcoidosis
[0347] In some embodiments, the present invention provides a method
of treating sarcoidosis comprising administering an effective
amount of an oxidative agent, including but not limited to chlorite
and chlorite-containing agents. In some embodiments, the present
invention provides a method of treating sarcoidosis comprising
administering an effective amount of an immunomodulatory agent,
including but not limited to immunosuppressive agents. In some
embodiments, an oxidative agent and an immunomodulatory agent are
used together for combination therapy to treat sarcoidosis.
[0348] Sarcoidosis, also called sarcoid or Besnier-Boeck disease,
is a multisystem disorder characterized by non-caseating granulomas
(small inflammatory nodules). Virtually any organ can be affected;
however, granulomas most often appear in the lungs or the lymph
nodes. Sarcoidosis has a paradoxical effect on inflammatory
processes; it is characterized by increased macrophage and CD4
helper T-cell activation resulting in accelerated inflammation,
however, immune responses to antigen challenges such as tuberculin
are suppressed. This paradoxic state of simultaneous hyper- and
hypo-activity is suggestive of a state of anergy. The anergy may
also be responsible for the increased risk of infections and
cancer. It appears that regulatory T-lymphocytes in the periphery
of sarcoid granulomas suppress IL-2 secretion which is hypothesized
to cause the state of anergy by preventing antigen-specific memory
responses (Kettritz R, et al. 2006 Nephrol. Dial. Transplant. 21
(10): 2690-4). While it is widely believed that TNF-alpha plays an
important role in the formation of granulomas it was observed that
sarcoidosis can be triggered by treatment with the TNF-alpha
antagonist etanercept (Verschueren K, et al. 2007 Clin. Rheumatol.
26 (11): 1969-71).
[0349] Sarcoidosis frequently causes a dysregulation of vitamin D
production with an increase in extrarenal production (Barbour G L,
et al. N Engl J Med 1981 305:440-43). Specifically, macrophages
inside the granulomas convert vitamin D to its active form,
resulting in elevated levels of the hormone 1,25-dihydroxyvitamin D
and symptoms of hypervitaminosis D that may include fatigue, lack
of strength or energy, irritability, metallic taste, temporary
memory loss or cognitive problems. Hypercalcemia (high calcium
levels) and its symptoms may be the result of excessive conversion
of vitamin D to its active form by epithelioid macrophages.
[0350] In some embodiments, the present invention provides a method
of treating sarcoidosis by modulating macrophage accumulation or
activation with an oxidative agent, including but not limited to
chlorite and chlorite-containing agents. In some embodiments, the
present invention provides a method of treating sarcoidosis by
modulating macrophage accumulation or activation with an
immunomodulatory agent, including but not limited to
immunosuppressive agents. In some embodiments, the macrophage
activation is reduced or inhibited.
[0351] XIII. Periodontitis
[0352] In some embodiments, the macrophage related disease that can
be treated with the method of the present invention is
periodontitis. Periodontitis typically refers to a number of
inflammatory diseases affecting the periodontium--that is, the
tissues that surround and support the teeth. Periodontitis involves
progressive loss of the alveolar bone around the teeth, and if left
untreated, can lead to the loosening and subsequent loss of teeth.
In periodontitis, overgrowth of Gram-negative bacteria and access
of lipopolysaccharide (LPS) to circulation may activate macrophages
leading to foam cell formation. Macrophages are the important
immune cells that are prominent at inflammatory periodontal sites.
It has been shown that the infected/inflamed area in periodontitis
is associated with macrophage activation via increased serum LPS
concentration (Pussinen, P J et al. Arteriosclerosis, Thrombosis,
and Vascular Biology 2004; 24:2174-2180). Furthermore, an expansion
of CD14+ CD16+ monocytes has been observed in periodontitis
(Nagasawa T, et al. J. Periodontal Res. 39, 72-78).
[0353] In some embodiments, the present invention provides a method
of treating periodontitis by modulating macrophage accumulation or
activation with an oxidative agent, including but not limited to
chlorite and chlorite-containing agents. In some embodiments, the
present invention provides a method of treating periodontitis by
modulating macrophage accumulation or activation with an
immunomodulatory agent, including but not limited to
immunosuppressive agents. In some embodiments, the macrophage
activation is reduced or inhibited. In some embodiments, an
oxidative agent and an immunomodulatory agent are used together for
combination therapy to treat periodontitis.
[0354] XIV. Whipple's Disease (WD)
[0355] In some embodiments, the present invention provides a method
of treating Whipple's disease comprising administering an effective
amount of an oxidative agent, including but not limited to chlorite
or a chlorite-containing compound, or a derivative of any thereof.
In some embodiments, the present invention provides a method of
treating Whipple's disease comprising administering an effective
amount of an immunomodulatory agent, including but not limited to
an immunosuppressive agent. In some embodiments, an oxidative agent
and an immunomodulatory agent are used together for combination
therapy to treat Whipple's disease.
[0356] Whipple's disease is a rare, systemic infectious disease
caused by the bacterium Tropheryma whipplei (Puechal X (2002).
Joint Bone Spine 69 (2): 133-40). Diagnosis is made by intestinal
biopsy, which reveals presence of the organism as Periodic
acid-Schiff (PAS)-positive macrophage inclusions. Intestinal
macrophages can induce an effective innate response and they
restrict inflammation via anergy. During WD, the small intestinal
mucosa of most patients is characterized by a loss of microvilli
and the infiltration of large foamy macrophages, which are filled
with PAS-positive material. These macrophages contained numerous
intracytoplasmic PAS-positive granules. A model has been proposed
that T. whipplei organisms are engulfed by resident intestinal
macrophages, which then shift toward the M2/alternatively activated
phenotype. They produce high levels of CCL18, IL-10, and TYRO
binding protein (DAP12), which may attract other macrophages and
naive T cells and orient the local immune response toward a Th2
response. Newly recruited macrophages engulf bacteria and produce
IL-16 and IL-1.beta. and undergo apoptosis. T whipplei infection
can then spread gradually (Desnues B. et al. Clin Vaccine Immunol.
2006 February; 13(2): 170-178).
[0357] In some embodiments, the present invention provides a method
of treating Whipple's disease by modulating macrophage activation
with an oxidative agent, including but not limited to chlorite and
chlorite-containing agents. In some embodiments, the present
invention provides a method of treating Whipple's disease by
modulating macrophage activation with an immunomodulatory agent,
including but not limited to immunosuppressive agents. In some
embodiments, the macrophage is intestinal macrophage.
[0358] XV. Pulmonary Alveolar Proteinosis
[0359] In some embodiments, the present invention provides a method
of treating pulmonary alveolar proteinosis comprising administering
an effective amount of an oxidative agent, including but not
limited to chlorite and chlorite-containing agents. In some
embodiments, the present invention provides a method of treating
pulmonary alveolar proteinosis comprising administering an
effective amount of an immunomodulatory agent, including but not
limited to immunosuppressive agents. In some embodiments, an
oxidative agent and an immunomodulatory agent are used together for
combination therapy to treat pulmonary alveolar proteinosis.
[0360] Pulmonary alveolar proteinosis (PAP) is a rare lung disease
in which abnormal accumulation of surfactant occurs within the
alveoli, interfering with gas exchange. PAP can occur in a primary
form or secondarily in the settings of malignancy (especially in
myeloid leukemia), pulmonary infection, or environmental exposure
to dusts or chemicals. Lung macrophages obtained by segmental
lavage from patients with pulmonary alveolar proteinosis have been
shown to exhibit morphologic abnormalities including excessive
lipid accumulation and giant secondary lysosome formation. These
cells survive poorly in tissue culture, show impaired chemotactic
activity, and have decreased adhesiveness to glass. They
phagocytize normally but have substantially decreased capacity to
kill ingested Candida pseudotropicalis. Previous studies suggest
that the lung macrophage in alveolar proteinosis is a defective
cell as a consequence of an abnormal pulmonary environment (Golde D
W et al. Annals of Internal Medicine 1976 85:304-309).
[0361] In some embodiments, the present invention provides a method
of treating pulmonary alveolar proteinosis by modulating macrophage
activation with an oxidative agent, including but not limited to
chlorite and chlorite-containing agents. In some embodiments, the
present invention provides a method of treating pulmonary alveolar
proteinosis comprising administering an effective amount of an
immunomodulatory agent, including but not limited to
immunosuppressive agents.
[0362] XVI. Macrophage-Related Pulmonary Diseases
[0363] In some embodiments, the present invention provides a method
of treating macrophage-related pulmonary diseases comprising
administering an effective amount of an oxidative agent, including
but not limited to chlorite and chlorite-containing agents. In some
embodiments, the present invention provides a method of treating
macrophage-related pulmonary diseases comprising administering an
effective amount of an immunomodulatory agent, including but not
limited to immunosuppressive agents. In some embodiments, an
oxidative agent and an immunomodulatory agent are used together for
combination therapy to treat macrophage-related pulmonary
diseases.
[0364] Macrophage-related pulmonary diseases are a heterogeneous
group of disorders characterized by macrophage accumulation or
activation. These conditions include without limitation
smoking-related interstitial lung diseases, metabolic disorders
such as Niemann-Pick or Gaucher's disease, and rare primary lung
tumors. High-resolution computed tomography abnormalities include
pulmonary ground-glass opacification secondary to infiltration by
macrophages, centrilobular nodules or interlobular septal
thickening reflecting peribronchiolar or septal macrophage
accumulation, respectively, emphysema caused by macrophage
dysfunction, and honeycombing following macrophage-related lung
matrix remodeling.
[0365] Niemann-Pick disease, also known as sphingomyelin lipidosis,
comprises a group of disorders characterized by foam cell
infiltration of the reticuloendothelial system. In the classic
infantile type A variant, a missense mutation causes complete
deficiency of sphingomyelinase. Sphingomyelin is a component of
cell membrane including the organellar membrane and so the enzyme
deficiency blocks degradation of lipid, resulting in the
accumulation of sphingomyelin within lysosomes in the
macrophage-monocyte phagocyte system. Histology demonstrates lipid
laden macrophages in the marrow, as well as "sea-blue histiocytes"
on pathology. Numerous small vacuoles of relatively uniform size
are created, imparting a foamness to the cytoplasm. Foam cells in
Niemann-Pick become engorged with sphingomyelin and, to a lesser
extent, other membrane lipids including cholesterol. Niemann-Pick
is typically caused by inactivation of the enzyme sphingomyelinase
in Type A and B disease, with 27-fold more residual enzyme activity
in Type B. Mutations in the sphingomyelin phosphodiesterase 1, acid
lysosomal (acid sphingomyelinase), also known as SMPD1 gene, cause
Niemann-Pick disease types A and B, and mutations in NPC1 and NPC2
cause Niemann-Pick disease, type C (NPC). NPC1 gene encodes a
putative integral membrane protein containing sequence motifs
consistent with a role in intracellular transport of cholesterol to
post-lysosomal destinations.
[0366] Gaucher's disease is one of the most common lysosomal
storage diseases. It is caused by a hereditary deficiency of the
enzyme glucocerebrosidase (also known as acid .beta.-glucosidase).
The enzyme acts on a fatty substance glucocerebroside (also known
as glucosylceramide, GlcCer). Glucocerebrosidase normally catalyzes
the hydrolysis of glucocerebroside to glucose and ceramide. When
the enzyme is defective, glucocerebroside accumulates, particularly
in cells of the mononuclear cell lineage, e.g., macrophages. Fatty
material can collect in the spleen, liver, kidneys, lungs, brain
and bone marrow. The macrophages that clear these cells are unable
to eliminate the waste product, which accumulates in fibrils, and
turn into Gaucher cells, which appear on light microscopy to
resemble crumpled-up paper. Diagnosis can be implied by biochemical
abnormalities such as high alkaline phosphatase,
angiotensin-converting enzyme (ACE) and immunoglobulin levels, or
by cell analysis showing "crinkled paper" cytoplasm and
glycolipid-laden macrophages.
[0367] In some embodiments, the present invention provides a method
of treating macrophage-related pulmonary diseases, including but
not limited to Niemann-Pick disease and Gaucher's disease, by
modulating macrophage accumulation or activation with an oxidative
agent, including but not limited to chlorite and
chlorite-containing agents. In some embodiments, the present
invention provides a method of treating macrophage-related
pulmonary diseases, including but not limited to Niemann-Pick
disease and Gaucher's disease, by modulating macrophage
accumulation or activation with an immunomodulatory agent,
including but not limited to immunosuppressive agents.
[0368] XVII. Leishmaniasis
[0369] In some embodiments, the present invention provides a method
of treating Leishmaniasis by modulating macrophage activation or
function with an oxidative agent, including but not limited to
chlorite and chlorite-containing agents. In some embodiments, the
present invention provides a method of treating Leishmaniasis by
modulating macrophage activation or function with an
immunomodulatory agent, including but not limited to
immunosuppressive agents. In some embodiments, an oxidative agent
and an immunomodulatory agent are used together for combination
therapy to treat Leishmaniasis.
[0370] Leishmaniasis is a disease caused by protozoan parasites
that belong to the genus Leishmania. Most forms of the disease are
transmissible only from animals (zoonosis), but some can be spread
between humans. Leishmaniasis is transmitted by the bite of female
phlebotomine sandflies. The sandflies inject the infective stage,
metacyclic promastigotes, during blood meals. Metacyclic
promastigotes that reach the puncture wound are phagocytized by
macrophages and transform into amastigotes. Amastigotes multiply in
infected cells and affect different tissues, depending in part on
which Leishmania species is involved. These differing tissue
specificities cause the differing clinical manifestations of the
various forms of leishmaniasis. Sandflies become infected during
blood meals on an infected host when they ingest macrophages
infected with amastigotes. In the sandfly's midgut, the parasites
differentiate into promastigotes, which multiply, differentiate
into metacyclic promastigotes and migrate to the proboscis.
Amastigotes are seen with monocytes or, less commonly in neutrophil
in peripheral blood and in macrophages in aspirates. In terms of
treatment, one study has reported that the intracellular growth of
Leishmania parasites was controlled by nelfinavir and ritonavir in
a human monocyte cell line and also in human primary
monocyte-derived macrophages (Trudel N. et al. (2008) Journal of
Infectious Diseases 198 (9): 1292-1299).
[0371] In some embodiments, the present invention provides a method
of treating Leishmaniasis by modulating macrophage activation or
function with an oxidative agent, including but not limited to
chlorite and chlorite-containing agents. In some embodiments, the
present invention provides a method of treating Leishmaniasis by
modulating macrophage activation or function with an
immunomodulatory agent, including but not limited to
immunosuppressive agents.
[0372] XVIII. Obesity
[0373] In some embodiments, the present invention provides a method
of treating obesity-related complications comprising administering
an effective amount of an oxidative agent to modulate macrophage
accumulation or activation. The oxidative agents include but are
not limited to chlorite and chlorite-containing compounds. In some
embodiments, the present invention provides a method of treating
obesity-related complications comprising administering an effective
amount of an immunomodulatory agent to modulate macrophage
accumulation or activation. In some embodiments, an oxidative agent
and an immunomodulatory agent are used together for combination
therapy to treat obesity-related complications.
[0374] Obesity alters adipose tissue metabolic and endocrine
function and leads to an increased release of fatty acids,
hormones, and proinflammatory molecules that contribute to obesity
associated complications. It has been demonstrated that obesity is
associated with increased macrophage infiltration into adipose
tissue and these adipose tissue macrophages participate in
inflammatory pathways that are activated in adipose tissues of
obese individuals. (Stuart P. Weisberg et al. Clin. Invest.
112(12): 1796-1808).
[0375] XIX. Hemodialysis
[0376] In some embodiments, the present invention provides a method
of treating or preventing post-hemodialysis inflammation comprising
administering an effective amount of an oxidative agent to modulate
macrophage accumulation or activation. The oxidative agents include
but are not limited to chlorite and chlorite-containing compounds.
In some embodiments, the present invention provides a method of
treating or preventing post-hemodialysis inflammation comprising
administering an effective amount of an immunomodulatory agent to
modulate macrophage accumulation or activation. In some
embodiments, an oxidative agent and an immunomodulatory agent are
used together for combination therapy to treat post-hemodialysis
inflammation.
[0377] Hemodialysis and extracorporal cardiopulmonary bypass
systems are based on direct contact of patient blood with
artificial membranes to allow for clearance of toxic metabolites
and for gas exchange, respectively. Exposure of leukocytes to such
membrane can induce an inflammatory response with increased
cytokine production. An increased number of CD14+ CD16+ monocytes
in hemodialysis patients have been observed (Saionji K, 2001 Acta
Haematol 105, 21-26). Adhesion of such monocytes to the membrane
and cytokine release by these monocytes may contribute to the
post-hemodialysis inflammatory syndrome (Caglar K, et al. Kidney
Int. 62, 1408-1416). In some embodiments, the oxidative agent,
including but not limited to chlorite and chlorite-containing
agents, is administered to a subject to treat or prevent
hemodialysis related inflammations by modulating macrophage
adhesion and activation. In some embodiments, the immunomodulatory
agent, including but not limited to immunosuppressive agents, is
administered to a subject to treat or prevent hemodialysis related
inflammations by modulating macrophage adhesion and activation.
[0378] XX. Microbial Infection
[0379] In some embodiments, the present invention provides a method
of treating microbial infection comprising administering an
effective amount of an oxidative agent to modulate macrophage
activation. In some embodiments, the present invention provides a
method of treating microbial infection comprising administering an
effective amount of an immunomodulatory agent to modulate
macrophage activation. In some embodiments, an oxidative agent and
an immunomodulatory agent are used together for combination therapy
to treat microbial infection.
[0380] Macrophages are the resident immune cells of the lung,
sensing bacteria and initiating subsequent host responses.
Following phagocytosis, macrophages recruit activated inflammatory
cells to the site of infection, as well as processing and
presenting bacterial antigens, thereby linking innate and adaptive
immunity Macrophages play an important part in the host defense
against infection because they are capable of phagocytosing
pathogens and their Fc.gamma. receptors recognize antibody-coated
bacteria. Natural killer cell activation also primes macrophages to
clear bacterial infection. Furthermore, pronounced increases of
CD14+ CD16+ monocytes have been observed in patients with a variety
of bacterial infections including bacterial sepsis, bacteremia,
neonatal sepsis, erysipelas, a skin infection, and hemolytic uremic
syndrome.
[0381] In some embodiments, the present invention provides a method
of treating microbial infection by modulating macrophage activation
or function with an oxidative agent, including but not limited to
chlorite and chlorite-containing agents. In some embodiments, the
present invention provides a method of treating microbial infection
by modulating macrophage activation or function with an
immunomodulatory agent, including but not limited to
immunosuppressive agents.
[0382] XXI. HIV Infection
[0383] In some embodiments, the present invention provides a method
of treating HIV infection comprising administering an effective
amount of an oxidative agent, including but not limited to chlorite
and chlorite-containing agents, to modulate macrophage activation.
In some embodiments, the present invention provides a method of
treating HIV infection comprising administering an effective amount
of an immunomodulatory agent, including but not limited to
immunosuppressive agents, to modulate macrophage activation. In
some embodiments, an oxidative agent and an immunomodulatory agent
are used together for combination therapy to treat HIV
infection.
[0384] Many viruses have been shown to modulate emerging immune
responses by infecting monocyte-macrophages. For example, monocytes
have been demonstrated to be productively infected by human
immunodeficiency virus (HIV) in vivo (Lambotte O, et al. J. Acquir.
Immune Defic. Syndr. 23, 114-119). The innate immune functions of
monocyte-macrophages, such as recruitment, differentiation and
chemokine secretion or activation of adaptive responses through
cytokine secretion, may alter the capacity of HIV-1 to infect and
replicate in macrophages. In addition to the central role of
macrophages in contributing to the immunopathology leading to AIDS,
macrophage infection in the CNS is directly associated with
viral-induced neuropathology. HIV-infected macrophages induce a
pro-inflammatory response when interacting with endothelial cells
(Pereira, C. F., et al. (2000) J. Leukoc. Biol. 68, 423-428) and
induce apoptosis in bystander astrocytes (Aquaro, S., et al. (2000)
J. Leukoc. Biol. 68, 429-435).
[0385] Results from clinical and non-clinical studies suggest that
the chlorite-containing agent WF10 may strengthen the patient's
native immunity against a variety of opportunistic infections and
malignancies, and inhibit the adverse effects of inflammatory
cytokines, which are stimulated as a result of HIV infection. See
McGrath et al., Development of WF10, a novel macrophage-regulating
agent, Curr Opin Investig Drugs, 3(3):365-73 (March 2002). Despite
stable antiretroviral therapy, patients with late-stage HIV
infection are less likely to respond to new antiretroviral drugs
and, thus, have few other treatment options. These patients
progress with diseases that are normally controlled by proper
macrophage function, e.g., opportunistic infections such as
cytomegalovirus (CMV) and Epstein-Barr virus (EBV). A small,
double-blind, placebo-controlled trial has indicated the potential
clinical benefit of WF10 on HIV infection. The rate of OIs and
hospitalizations was significantly lower in the WF10 group than in
the placebo group and long-term follow-up suggested a survival
benefit to WF 10-treated patients with HIV infection.
[0386] Regulation of macrophage function with oxidative agents may
provide a treatment option that is not dependent upon inhibition of
virus replication. Administered in conjunction with HAART,
oxidative agents, including but not limited to chlorite or
chlorite-containing agents, can enhance treatment effectiveness in
the management of HIV disease by restoring immune function through
regulation of macrophage function to downregulate inappropriate
T-cell activation Immunomodulatory agents can be used to provide
similar effects. In some embodiments, oxidative and
immunomodulatory agents are used together to enhance treatment
effectiveness in HIV. On the basis of the balanced macrophage
activation model of human disease, over time, such oxidative agents
could partially restore normal macrophage function to even advanced
AIDS patients. Clinical manifestations of poor macrophage function,
such as pneumocystis pneumonia and mycobacterial infections
normally cleared by phagocytosis should decrease, normal macrophage
activation of appropriate T-cell function, although limited, may
prevent recurrence of viral diseases (such as EBV lymphoma or CMV
retinitis), and anti-inflammatory activity may decrease the
frequency of AIDS-related dementia and wasting syndrome.
Considering that macrophages are longlived cells with defined
long-term baseline functional characteristics, oxidative
agent-mediated re-establishment of normal baseline function may
persist long after the actual period of drug administration.
[0387] In some embodiments, an oxidative agent, including but not
limited to chlorite or chlorite-containing agents, is administered
in a subject to treat microbial infections including bacterial
infections and viral infections by modulating macrophage activation
or function. In some embodiments, an immunomodulatory agent,
including immunostimulating and immunosuppressive agents, is
administered in a subject to treat microbial infections including
bacterial infections and viral infections by modulating macrophage
activation or function. In some embodiments, the viral infection
comprises HIV. In some embodiments, the macrophage activation is
enhanced. In some embodiments, the macrophage activation is
inhibited.
[0388] XXII. Cancer
[0389] In some embodiments, the present invention provides a method
of treating cancer comprising administering an effective amount of
an oxidative agent, including but not limited to chlorite or
chlorite-containing agents, to modulate macrophage activation. In
some embodiments, the present invention provides a method of
treating cancer comprising administering an effective amount of an
immunomodulatory agent to modulate macrophage activation. In some
embodiments, an oxidative agent and an immunomodulatory agent are
used together for combination therapy to treat cancer.
[0390] Cancer (malignant neoplasm) is a class of diseases in which
a group of cells display uncontrolled growth, invasion, and
sometimes metastasis to other locations in the body via lymph or
blood. These three malignant properties of cancers differentiate
them from benign tumors, which are self-limited, and do not invade
or metastasize.
[0391] The role of macrophages in tumor growth and development is
complex and multifaceted. Although there is limited evidence that
tumor-associated macrophages (TAMs) can be directly tumoricidal and
stimulate the anti-tumor activity of T cells, there is contrasting
evidence that tumor cells are able to block or evade the activity
of TAMs at the tumor site. In some cases, tumor-derived molecules
even redirect TAM activities to promote tumor survival and growth.
Evidence has emerged for a symbiotic relationship between tumor
cells and TAMs, in which tumor cells attract TAMs and sustain their
survival, with TAMs then responding to micro-environmental factors
in tumors such as hypoxia by producing important mitogens as well
as various growth factors and enzymes that stimulate tumor
angiogenesis.
[0392] Macrophages are generally not tumoricidal for tumor cells
unless activated, for example, by antibodies or classic macrophages
stimulants such as IFN-y or lipopolysaccharide LPS. Once activated,
direct cytotoxicity is exerted towards tumor cells, or indirect
cytotoxicity is exerted via the secretion of factors that stimulate
the anti-tumor functions of other cell types. Direct cytotoxicity
can be further divided into macrophage-mediated tumor cytotoxicity
and antibody-dependent cellular cytotoxicity. IFN-y produced by T
cells and NK cells induces macrophages to release reactive oxygen
and nitrogen species that have anti-tumor effects. On the other
hand, TAM may also have pro-tumor effects. For example, TAM-derived
cytokines may stimulate tumor cells to produce angiogenin,
resulting in tumor angiogenesis (Bingle L, et al. Journal of
Pathology, 2002 196, 254-265). Moreover, TAM accumulation in
hypoxic tumor areas has important implications. Human macrophages
have been shown to respond to environmental hypoxia with an
increased release of VEGF, which is a key component of the
angiogenic process in a variety of human tumors (Ferrara N. Tumor
Angiogenesis, Oxford University Press: Oxford, 1997 185-199; Lewis
J S et al. J Leukoc Biol 1999 66:889-900).
[0393] Given that tumor cells attract macrophages and sustain their
survival and TAMs produce a myriad of factors to promote tumor
growth and angiogenesis, activated or genetically modified
macrophages may be used to restrain tumor growth. Moreover,
macrophage products such as cytokines with anti-tumor effects may
have utility in the treatment of cancer. In some embodiments, an
oxidative agent such as chlorite is administered to a subject to
treat cancer by modulating macrophage activation or function. In
some embodiments, an immunomodulatory agent is administered to a
subject to treat cancer by modulating macrophage activation or
function.
[0394] The types of cancer that can be treated using the methods of
the present invention include but are not limited to adrenal
cortical cancer, anal cancer, aplastic anemia, bile duct cancer,
bladder cancer, bone cancer, bone metastasis, central nervous
system (CNS) cancers, peripheral nervous system (PNS) cancers,
breast cancer, Castleman's Disease, cervical cancer, childhood
Non-Hodgkin's lymphoma, colon and rectum cancer, endometrial
cancer, esophagus cancer, Ewing's family of tumors (e.g., Ewing's
sarcoma), eye cancer, gallbladder cancer, gastrointestinal
carcinoid tumors, gastrointestinal stromal tumors, gestational
trophoblastic disease, hairy cell leukemia, Hodgkin's disease,
Kaposi's sarcoma, kidney cancer, laryngeal and hypopharyngeal
cancer, acute lymphocytic leukemia, acute myeloid leukemia,
children's leukemia, chronic lymphocytic leukemia, chronic myeloid
leukemia, liver cancer, lung cancer, lung carcinoid tumors,
Non-Hodgkin's lymphoma, male breast cancer, malignant mesothelioma,
multiple myeloma, myelodysplastic syndrome, myeloproliferative
disorders, nasal cavity and paranasal cancer, nasopharyngeal
cancer, neuroblastoma, oral cavity and oropharyngeal cancer,
osteosarcoma, ovarian cancer, pancreatic cancer, penile cancer,
pituitary tumor, prostate cancer, retinoblastoma, rhabdomyosarcoma,
salivary gland cancer, sarcoma (adult soft tissue cancer), melanoma
skin cancer, non-melanoma skin cancer, stomach cancer, testicular
cancer, thymus cancer, thyroid cancer, uterine cancer (e.g.,
uterine sarcoma), vaginal cancer, vulvar cancer, and Waldenstrom's
macroglobulinemia.
[0395] In some embodiments, the cancer comprises Acute
Lymphoblastic Leukemia. In other embodiments, the cancer comprises
Acute Myeloid Leukemia. In other embodiments, the cancer comprises
Adrenocortical Carcinoma. In other embodiments, the cancer
comprises an AIDS-Related Cancer. In other embodiments, the cancer
comprises AIDS-Related Lymphoma. In other embodiments, the cancer
comprises Anal Cancer. In other embodiments, the cancer comprises
Appendix Cancer. In other embodiments, the cancer comprises
Childhood Cerebellar Astrocytoma. In other embodiments, the cancer
comprises Childhood Cerebral Astrocytoma. In other embodiments, the
cancer comprises a Central Nervous System Atypical
Teratoid/Rhabdoid Tumor. In other embodiments, the cancer comprises
Basal Cell Carcinoma, or other Skin Cancer (Nonmelanoma). In other
embodiments, the cancer comprises Extrahepatic Bile Duct Cancer. In
other embodiments, the cancer comprises Bladder Cancer. In other
embodiments, the cancer comprises Bone Cancer, such as Osteosarcoma
or Malignant Fibrous Histiocytoma. In other embodiments, the cancer
comprises Brain Stem Glioma. In other embodiments, the cancer
comprises an Adult Brain Tumor. In other embodiments, the cancer
comprises Brain Tumor, Central Nervous System Atypical
Teratoid/Rhabdoid Tumor, Childhood. In other embodiments, the
cancer comprises a Brain Tumor comprising Cerebral
Astrocytoma/Malignant Glioma. In other embodiments, the cancer
comprises a Craniopharyngioma Brain Tumor. In other embodiments,
the cancer comprises a Ependymoblastoma Brain Tumor. In other
embodiments, the cancer comprises a Ependymoma Brain Tumor. In
other embodiments, the cancer comprises a Medulloblastoma Brain
Tumor. In other embodiments, the cancer comprises a
Medulloepithelioma Brain Tumor. In other embodiments, the cancer
comprises Brain Tumors including Pineal Parenchymal Tumors of
Intermediate Differentiation. In other embodiments, the cancer
comprises Brain Tumors including Supratentorial Primitive
Neuroectodermal Tumors and Pineoblastoma. In other embodiments, the
cancer comprises a Brain Tumor including Visual Pathway and
Hypothalamic Glioma. In other embodiments, the cancer comprises
Brain and Spinal Cord Tumors. In other embodiments, the cancer
comprises Breast Cancer. In other embodiments, the cancer comprises
Bronchial Tumors. In other embodiments, the cancer comprises
Burkitt Lymphoma. In other embodiments, the cancer comprises
Carcinoid Tumor. In other embodiments, the cancer comprises
Gastrointestinal Carcinoid Tumor. In other embodiments, the cancer
comprises Carcinoma of Unknown Primary Origin. In other
embodiments, the cancer comprises Central Nervous System Atypical
Teratoid/Rhabdoid Tumor. In other embodiments, the cancer comprises
Central Nervous System Embryonal Tumors. In other embodiments, the
cancer comprises Primary Central Nervous System Lymphoma. In other
embodiments, the cancer comprises Cerebellar Astrocytoma. In other
embodiments, the cancer comprises Cerebral Astrocytoma/Malignant
Glioma. In other embodiments, the cancer comprises Cervical Cancer.
In other embodiments, the cancer comprises Childhood Cancers. In
other embodiments, the cancer comprises Chordoma. In other
embodiments, the cancer comprises Chronic Lymphocytic Leukemia. In
other embodiments, the cancer comprises Chronic Myelogenous
Leukemia. In other embodiments, the cancer comprises Chronic
Myeloproliferative Disorders. In other embodiments, the cancer
comprises Colon Cancer. In other embodiments, the cancer comprises
Colorectal Cancer. In other embodiments, the cancer comprises
Craniopharyngioma. In other embodiments, the cancer comprises
Cutaneous T-Cell Lymphoma, including Mycosis Fungoides and Sezary
Syndrome. In other embodiments, the cancer comprises Central
Nervous System Embryonal Tumors. In other embodiments, the cancer
comprises Endometrial Cancer. In other embodiments, the cancer
comprises Ependymoblastoma. In other embodiments, the cancer
comprises Ependymoma. In other embodiments, the cancer comprises
Esophageal Cancer. In other embodiments, the cancer comprises the
Ewing Family of Tumors. In other embodiments, the cancer comprises
Extracranial Germ Cell Tumor. In other embodiments, the cancer
comprises Extragonadal Germ Cell Tumor. In other embodiments, the
cancer comprises Extrahepatic Bile Duct Cancer. In other
embodiments, the cancer comprises Intraocular Melanoma Eye Cancer.
In other embodiments, the cancer comprises Retinoblastoma Eye
Cancer. In other embodiments, the cancer comprises Gallbladder
Cancer. In other embodiments, the cancer comprises Gastric
(Stomach) Cancer. In other embodiments, the cancer comprises
Gastrointestinal Carcinoid Tumor. In other embodiments, the cancer
comprises Gastrointestinal Stromal Tumor (GIST). In other
embodiments, the cancer comprises Gastrointestinal Stromal Cell
Tumor. In other embodiments, the cancer comprises Extracranial Germ
Cell Tumor. In other embodiments, the cancer comprises Extragonadal
Germ Cell Tumor. In other embodiments, the cancer comprises Ovarian
Germ Cell Tumor. In other embodiments, the cancer comprises
Gestational Trophoblastic Tumor. In other embodiments, the cancer
comprises Glioma. In other embodiments, the cancer comprises Brain
Stem Glioma. In other embodiments, the cancer comprises Cerebral
Astrocytoma Glioma. In other embodiments, the cancer comprises
Visual Pathway or Hypothalamic Glioma. In other embodiments, the
cancer comprises Hairy Cell Leukemia. In other embodiments, the
cancer comprises Head and Neck Cancer. In other embodiments, the
cancer comprises Hepatocellular (Liver) Cancer. In other
embodiments, the cancer comprises Hodgkin Lymphoma. In other
embodiments, the cancer comprises Hypopharyngeal Cancer. In other
embodiments, the cancer comprises Intraocular Melanoma. In other
embodiments, the cancer comprises Islet Cell Tumors (Endocrine
Pancreas). In other embodiments, the cancer comprises Kaposi
Sarcoma. In other embodiments, the cancer comprises Kidney (Renal
Cell) Cancer. In other embodiments, the cancer comprises Laryngeal
Cancer. In other embodiments, the cancer comprises Acute
Lymphoblastic Leukemia. In other embodiments, the cancer comprises
Acute Myeloid Leukemia. In other embodiments, the cancer comprises
Chronic Lymphocytic Leukemia. In other embodiments, the cancer
comprises Chronic Myelogenous Leukemia. In other embodiments, the
cancer comprises Hairy Cell Leukemia. In other embodiments, the
cancer comprises Lip Cancer. In other embodiments, the cancer
comprises Oral Cavity Cancer. In other embodiments, the cancer
comprises Primary Liver Cancer. In other embodiments, the cancer
comprises Non-Small Cell Lung Cancer. In other embodiments, the
cancer comprises Small Cell Lung Cancer. In other embodiments, the
cancer comprises AIDS-Related Lymphoma. In other embodiments, the
cancer comprises Burkitt Lymphoma. In other embodiments, the cancer
comprises Cutaneous T-Cell Lymphoma. In other embodiments, the
cancer comprises Mycosis Fungoides and Sezary Syndrome. In other
embodiments, the cancer comprises Hodgkin Lymphoma. In other
embodiments, the cancer comprises Non-Hodgkin Lymphoma. In other
embodiments, the cancer comprises Primary Central Nervous System
Lymphoma. In other embodiments, the cancer comprises Waldenstrom
Macroglobulinemia. In other embodiments, the cancer comprises
Malignant Fibrous Histiocytoma of Bone or Osteosarcoma. In other
embodiments, the cancer comprises Medulloepithelioma. In other
embodiments, the cancer comprises Melanoma. In other embodiments,
the cancer comprises Intraocular (Eye) Melanoma. In other
embodiments, the cancer comprises Merkel Cell Carcinoma. In other
embodiments, the cancer comprises Mesothelioma. In other
embodiments, the cancer comprises Metastatic Squamous Neck Cancer
with Occult Primary. In other embodiments, the cancer comprises
Mouth Cancer. In other embodiments, the cancer comprises Multiple
Endocrine Neoplasia Syndrome. In other embodiments, the cancer
comprises Multiple Myeloma/Plasma Cell Neoplasm. In other
embodiments, the cancer comprises Mycosis Fungoides. In other
embodiments, the cancer comprises Myelodysplastic Syndromes. In
other embodiments, the cancer comprises Myelodysplastic or
Myeloproliferative Diseases. In other embodiments, the cancer
comprises Chronic Myelogenous Leukemia. In other embodiments, the
cancer comprises Acute Myeloid Leukemia. In other embodiments, the
cancer comprises Multiple Myeloma. In other embodiments, the cancer
comprises Chronic Myeloproliferative Disorders. In other
embodiments, the cancer comprises Nasal Cavity or Paranasal Sinus
Cancer. In other embodiments, the cancer comprises Nasopharyngeal
Cancer. In other embodiments, the cancer comprises Nasopharyngeal
Cancer. In other embodiments, the cancer comprises Neuroblastoma.
In other embodiments, the cancer comprises Non-Hodgkin Lymphoma. In
other embodiments, the cancer comprises Non-Small Cell Lung Cancer.
In other embodiments, the cancer comprises Oral Cancer. In other
embodiments, the cancer comprises Oral Cavity Cancer. In other
embodiments, the cancer comprises Oropharyngeal Cancer. In other
embodiments, the cancer comprises Osteosarcoma. In other
embodiments, the cancer comprises Malignant Fibrous Histiocytoma of
Bone. In other embodiments, the cancer comprises Ovarian Cancer. In
other embodiments, the cancer comprises Ovarian Epithelial Cancer.
In other embodiments, the cancer comprises Ovarian Germ Cell Tumor.
In other embodiments, the cancer comprises Ovarian Low Malignant
Potential Tumor. In other embodiments, the cancer comprises
Pancreatic Cancer. In other embodiments, the cancer comprises Islet
Cell Tumor Pancreatic Cancer. In other embodiments, the cancer
comprises Papillomatosis. In other embodiments, the cancer
comprises Paranasal Sinus Cancer. In other embodiments, the cancer
comprises Nasal Cavity Cancer. In other embodiments, the cancer
comprises Parathyroid Cancer. In other embodiments, the cancer
comprises Penile Cancer. In other embodiments, the cancer comprises
Pharyngeal Cancer. In other embodiments, the cancer comprises
Pheochromocytoma. In other embodiments, the cancer comprises Pineal
Parenchymal Tumors of Intermediate Differentiation. In other
embodiments, the cancer comprises Pineoblastoma or Supratentorial
Primitive Neuroectodermal Tumors. In other embodiments, the cancer
comprises Pituitary Tumor. In other embodiments, the cancer
comprises Plasma Cell Neoplasm/Multiple Myeloma. In other
embodiments, the cancer comprises Pleuropulmonary Blastoma. In
other embodiments, the cancer comprises Primary Central Nervous
System Lymphoma. In other embodiments, the cancer comprises
Prostate Cancer. In other embodiments, the cancer comprises Rectal
Cancer. In other embodiments, the cancer comprises Renal Cell
(Kidney) Cancer. In other embodiments, the cancer comprises Renal
Pelvis and Ureter, Transitional Cell Cancer. In other embodiments,
the cancer comprises Respiratory Tract Carcinoma Involving the NUT
Gene on Chromosome 15. In other embodiments, the cancer comprises
Retinoblastoma. In other embodiments, the cancer comprises
Rhabdomyosarcoma. In other embodiments, the cancer comprises
Salivary Gland Cancer. In other embodiments, the cancer comprises
Sarcoma of the Ewing Family of Tumors. In other embodiments, the
cancer comprises Kaposi Sarcoma. In other embodiments, the cancer
comprises Soft Tissue Sarcoma. In other embodiments, the cancer
comprises Uterine Sarcoma. In other embodiments, the cancer
comprises Sezary Syndrome. In other embodiments, the cancer
comprises Nonmelanoma Skin Cancer. In other embodiments, the cancer
comprises Melanoma Skin Cancer. In other embodiments, the cancer
comprises Merkel Cell Skin Carcinoma. In other embodiments, the
cancer comprises Small Cell Lung Cancer. In other embodiments, the
cancer comprises Small Intestine Cancer. In other embodiments, the
cancer comprises Squamous Cell Carcinoma, e.g., Nonmelanoma Skin
Cancer. In other embodiments, the cancer comprises Metastatic
Squamous Neck Cancer with Occult Primary. In other embodiments, the
cancer comprises Stomach (Gastric) Cancer. In other embodiments,
the cancer comprises Supratentorial Primitive Neuroectodermal
Tumors. In other embodiments, the cancer comprises Cutaneous T-Cell
Lymphoma, e.g., Mycosis Fungoides and Sezary Syndrome. In other
embodiments, the cancer comprises Testicular Cancer. In other
embodiments, the cancer comprises Throat Cancer. In other
embodiments, the cancer comprises Thymoma or Thymic Carcinoma. In
other embodiments, the cancer comprises Thyroid Cancer. In other
embodiments, the cancer comprises Transitional Cell Cancer of the
Renal Pelvis and Ureter. In other embodiments, the cancer comprises
Gestational Trophoblastic Tumor. In other embodiments, the cancer
comprises a Carcinoma of Unknown Primary Site. In other
embodiments, the cancer comprises an Unusual Cancer of Childhood.
In other embodiments, the cancer comprises Ureter and Renal Pelvis
Transitional Cell Cancer. In other embodiments, the cancer
comprises Urethral Cancer. In other embodiments, the cancer
comprises Endometrial Uterine Cancer. In other embodiments, the
cancer comprises Uterine Sarcoma. In other embodiments, the cancer
comprises Vaginal Cancer. In other embodiments, the cancer
comprises Visual Pathway and Hypothalamic Glioma. In other
embodiments, the cancer comprises Vulvar Cancer. In other
embodiments, the cancer comprises Waldenstrom Macroglobulinemia. In
other embodiments, the cancer comprises Wilms Tumor. In other
embodiments, the cancer comprises Women's Cancers.
[0396] In some embodiments, the present invention provides a method
of treating cancer comprising administering an effective amount of
an oxidative agent, including but not limited to chlorite or
chlorite-containing agents. In some embodiments, the present
invention provides a method of treating cancer comprising
administering an effective amount of an immunomodulatory agent,
including but not limited to immunosuppressive agents. The
oxidative agent or immunomodulatory agent of the invention can be
used in combination with another anti-cancer therapy, including but
not limited to chemotherapy, surgery, radiation therapy (e.g., X
ray), gene therapy, immunotherapy, DNA therapy, adjuvant therapy,
neoadjuvant therapy, viral therapy, RNA therapy, and
nanotherapy.
[0397] (a) Breast Cancer
[0398] In one embodiment, the present invention provides a method
of treating breast cancer comprising administering an effective
amount of an oxidative agent to modulate macrophage activation. In
one embodiment, the present invention provides a method of treating
breast cancer comprising administering an effective amount of an
immunomodulatory agent to modulate macrophage activation. In some
embodiments, an oxidative agent and an immunomodulatory agent are
used together for combination therapy to treat breast cancer.
[0399] Macrophages may play an important role in breast cancer
development. Research performed with an MMTV-PyMT mouse model of
mammary carcinogenesis has revealed a tumor-promoting role for
TH2-CD4 T effector cells, suggesting the immune system modulates
the early onset of cancer development in specific organs, and
confirmed epidemiological studies showing that increased macrophage
presence correlates with higher tumor grade and decreased survival
(Rochman S. et al. BMC Proceedings 2009, 3(Suppl 5):I1).
Eliminating the T effector cells did not regulate primary disease,
and also made it more likely that the cancer would metastasize to
the lung, behavior that appeared to be regulated by
macrophages.
[0400] In one embodiment, the present invention provides a method
of treating breast cancer comprising administering an effective
amount of an oxidative agent, including but not limited to chlorite
or chlorite-containing agents. In one embodiment, the present
invention provides a method of treating breast cancer comprising
administering an effective amount of an immunomodulatory agent,
e.g., to modulate the activity of macrophages. The oxidative agent
or immunomodulatory agent of the invention can be used in
combination with a second anti-breast cancer therapy including but
not limited to chemotherapy, surgery, radiation therapy (e.g., X
ray), gene therapy, immunotherapy, DNA therapy, adjuvant therapy,
neoadjuvant therapy, viral therapy, RNA therapy, and
nanotherapy.
[0401] (b) Anti-hyperproliferative Agents
[0402] In some embodiments, the present invention provides a method
that comprises administering an oxidative agent and/or
immunomodulatory agent in combination with one or more other
therapeutic agents and/or interventions used for the treatment of
cancer or a hyperproliferative disease. The therapeutic agents that
can be used in combination with oxidative agent of the present
invention for the treatment of cancer or a hyperproliferative
disease include but are not limited to the following
categories.
[0403] (i) Antineoplastic chemotherapeutic agents
[0404] Suitable antineoplastic anti-tumor agents that can be used
in the present invention include, but are not limited to,
alkylating agents, antimetabolites, natural antineoplastic agents,
hormonal antineoplastic agents, angiogenesis inhibitors,
differentiating reagents, RNA inhibitors, antibodies or
immunotherapeutic agents, gene therapy agents, small molecule
enzymatic inhibitors, biological response modifiers, and
anti-metastatic agents.
[0405] (ii) Alkylating agents
[0406] Alkylating agents are known to act through the alkylation of
macromolecules such as the DNA of cancer cells, and are usually
strong electrophiles. This activity can disrupt DNA synthesis and
cell division. Examples of alkylating reagents suitable for use
herein include nitrogen mustards and their analogues and
derivatives including, cyclophosphamide, ifosfamide, chlorambucil,
estramustine, mechlorethamine hydrochloride, melphalan, and uracil
mustard. Other examples of alkylating agents include alkyl
sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine,
lomustine, and streptozocin), triazenes (e.g., dacarbazine and
temozolomide), ethylenimines/methylmelamines (e.g., altretamine and
thiotepa), and methylhydrazine derivatives (e.g., procarbazine).
Included in the alkylating agent group are the alkylating-like
platinum-containing drugs comprising carboplatin, cisplatin, and
oxaliplatin.
[0407] (iii) Antimetabolites
[0408] Antimetabolic antineoplastic agents structurally resemble
natural metabolites, and are involved in normal metabolic processes
of cancer cells such as the synthesis of nucleic acids and
proteins. They differ enough from the natural metabolites so that
they interfere with the metabolic processes of cancer cells.
Suitable antimetabolic antineoplastic agents that can be used in
the present invention can be classified according to the metabolic
process they affect, and can include, but are not limited to,
analogues and derivatives of folic acid, pyrimidines, purines, and
cytidine. Members of the folic acid group of agents suitable for
use herein include, but are not limited to, methotrexate
(amethopterin), pemetrexed and their analogues and derivatives.
Pyrimidine agents suitable for use herein include, but are not
limited to, cytarabine, floxuridine, fluorouracil (5-fluorouracil),
capecitabine, gemcitabine, and their analogues and derivatives.
Purine agents suitable for use herein include, but are not limited
to, mercaptopurine (6-mercaptopurine), pentostatin, thioguanine,
cladribine, and their analogues and derivatives. Cytidine agents
suitable for use herein include, but are not limited to, cytarabine
(cytosine arabinodside), azacitidine (5-azacytidine) and their
analogues and derivatives.
[0409] (iv) Natural antineoplastic agents
[0410] Natural antineoplastic agents comprise antimitotic agents,
antibiotic antineoplastic agents, camptothecin analogues, and
enzymes. Antimitotic agents suitable for use herein include, but
are not limited to, vinca alkaloids like vinblastine, vincristine,
vindesine, vinorelbine, and their analogues and derivatives. They
are derived from the Madagascar periwinkle plant and are usually
cell cycle-specific for the M phase, binding to tubulin in the
microtubules of cancer cells. Other antimitotic agents suitable for
use herein are the podophyllotoxins, which include, but are not
limited to etoposide, teniposide, and their analogues and
derivatives. These reagents predominantly target the G2 and late S
phase of the cell cycle.
[0411] Also included among the natural antineoplastic agents are
the antibiotic antineoplastic agents. Antibiotic antineoplastic
agents are antimicrobial drugs that have anti-tumor properties
usually through interacting with cancer cell DNA. Antibiotic
antineoplastic agents suitable for use herein include, but are not
limited to, belomycin, dactinomycin, doxorubicin, idarubicin,
epirubicin, mitomycin, mitoxantrone, pentostatin, plicamycin, and
their analogues and derivatives.
[0412] The natural antineoplastic agent classification also
includes camptothecin analogues and derivatives which are suitable
for use herein and include camptothecin, topotecan, and irinotecan.
These agents act primarily by targeting the nuclear enzyme
topoisomerase I. Another subclass under the natural antineoplastic
agents is the enzyme, L-asparaginase and its variants.
L-asparaginase acts by depriving some cancer cells of L-asparagine
by catalyzing the hydrolysis of circulating asparagine to aspartic
acid and ammonia.
[0413] (v) Hormonal antineoplastic agents
[0414] Hormonal antineoplastic agents act predominantly on
hormone-dependent cancer cells associated with prostate tissue,
breast tissue, endometrial tissue, ovarian tissue, lymphoma, and
leukemia. Such tissues may be responsive to and dependent upon such
classes of agents as glucocorticoids, progestins, estrogens, and
androgens. Both analogues and derivatives that are agonists or
antagonists are suitable for use in the present invention to treat
tumors. Examples of glucocorticoid agonists/antagonists suitable
for use herein are dexamethasone, cortisol, corticosterone,
prednisone, mifepristone (RU486), their analogues and derivatives.
The progestin agonist/antagonist subclass of agents suitable for
use herein includes, but is not limited to, hydroxyprogesterone,
medroxyprogesterone, megestrol acetate, mifepristone (RU486),
ZK98299, their analogues and derivatives. Examples from the
estrogen agonist/antagonist subclass of agents suitable for use
herein include, but are not limited to, estrogen, tamoxifen,
toremifene, RU58668, SR16234, ZD164384, ZK191703, fulvestrant,
their analogues and derivatives. Examples of aromatase inhibitors
suitable for use herein, which inhibit estrogen production,
include, but are not limited to, androstenedione, formestane,
exemestane, aminoglutethimide, anastrozole, letrozole, their
analogues and derivatives. Examples from the androgen
agonist/antagonist subclass of agents suitable for use herein
include, but are not limited to, testosterone, dihydrotestosterone,
fluoxymesterone, testolactone, testosterone enanthate, testosterone
propionate, gonadotropin-releasing hormone agonists/antagonists
(e.g., leuprolide, goserelin, triptorelin, buserelin),
diethylstilbestrol, abarelix, cyproterone, flutamide, nilutamide,
bicalutamide, their analogues and derivatives.
[0415] (vi) Angiogenesis inhibitors
[0416] Angiogenesis inhibitors work by inhibiting the
vascularization of tumors. Angiogenesis inhibitors encompass a wide
variety of agents including small molecule agents, antibody agents,
and agents that target RNA function. Examples of angiogenesis
inhibitors suitable for use herein include, but are not limited to,
ranibizumab, bevacizumab, SU11248, PTK787, ZK222584, CEP-7055,
angiozyme, dalteparin, thalidomide, suramin, CC-5013,
combretastatin A4 Phosphate, LY317615, soy isoflavones, AE-941,
interferon alpha, PTK787/ZK 222584, ZD6474, EMD 121974, ZD6474, BAY
543-9006, celecoxib, halofuginone hydrobromide, bevacizumab, their
analogues, variants, or derivatives.
[0417] (vii) Differentiating agents
[0418] Differentiating agents inhibit tumor growth through
mechanisms that induce cancer cells to differentiate. One such
subclass of these agents suitable for use herein includes, but is
not limited to, vitamin A analogues or retinoids, and peroxisome
proliferator-activated receptor agonists (PPARs). Retinoids
suitable for use herein include, but are not limited to, vitamin A,
vitamin A aldehyde (retinal), retinoic acid, fenretinide,
9-cis-retinoid acid, 13-cis-retinoid acid, all-trans-retinoic acid,
isotretinoin, tretinoin, retinal palmitate, their analogues and
derivatives. Agonists of PPARs suitable for use herein include, but
are not limited to, troglitazone, ciglitazone, tesaglitazar, their
analogues and derivatives.
[0419] (viii) Antibodies/immunotherapeutic agents
[0420] Antibody agents bind targets selectively expressed in cancer
cells and can either utilize a conjugate to kill the cell
associated with the target, or elicit the body's immune response to
destroy the cancer cells. Immunotherapeutic agents can either be
comprised of polyclonal or monoclonal antibodies. The antibodies
may be comprised of non-human animal (e.g., mouse) and human
components, or be comprised of entirely human components
("humanized antibodies"). Examples of monoclonal immunotherapeutic
agents suitable for use herein include, but are not limited to,
rituximab, tosibtumomab, ibritumomab which target the CD-20
protein. Other examples suitable for use herein include
trastuzumab, edrecolomab, bevacizumab, cetuximab, carcinoembryonic
antigen antibodies, gemtuzumab, alemtuzumab, mapatumumab,
panitumumab, EMD 72000, TheraCIM hR3, 2C4, HGS-TR2J, and
HGS-ETR2.
[0421] (ix) Gene therapy agents
[0422] Gene therapy agents insert copies of genes into a specific
set of a patient's cells, and can target both cancer and non-cancer
cells. The goal of gene therapy can be to replace altered genes
with functional genes, to stimulate a patient's immune response to
cancer, to make cancer cells more sensitive to chemotherapy, to
place "suicide" genes into cancer cells, or to inhibit
angiogenesis. Genes may be delivered to target cells using viruses,
liposomes, or other carriers or vectors. This may be done by
injecting the gene-carrier composition into the patient directly,
or ex vivo, with infected cells being introduced back into a
patient. Such compositions are suitable for use in the present
invention.
[0423] (x) Nanotherapy
[0424] Nanometer-sized particles have novel optical, electronic,
and structural properties that are not available from either
individual molecules or bulk solids. When linked with
tumor-targeting moieties, such as tumor-specific ligands or
monoclonal antibodies, these nanoparticles can be used to target
cancer-specific receptors, tumor antigens (biomarkers), and tumor
vasculatures with high affinity and precision. The formuation and
manufacturing process for cancer nanotherapy is disclosed in U.S.
Pat. No. 7,179,484, filed Nov. 6, 2003 and entitled
"Protein-stabilized liposomal formulations of pharmaceutical
agents," and article M. N. Khalid, P. Simard, D. Hoarau, A.
Dragomir, J. Leroux, Long Circulating Poly(Ethylene
Glycol)Decorated Lipid Nanocapsules Deliver Docetaxel to Solid
Tumors, Pharmaceutical Research, 23(4), 2006, all of which are
herein incorporated by reference in their entireties.
[0425] (xi) RNA therapy
[0426] RNA including but not limited to siRNA, shRNA, microRNA may
be used to modulate gene expression and treat cancers. Double
stranded oligonucleotides are formed by the assembly of two
distinct oligonucleotide sequences where the oligonucleotide
sequence of one strand is complementary to the oligonucleotide
sequence of the second strand; such double stranded
oligonucleotides are generally assembled from two separate
oligonucleotides (e.g., siRNA), or from a single molecule that
folds on itself to form a double stranded structure (e.g., shRNA or
short hairpin RNA). These double stranded oligonucleotides known in
the art all have a common feature in that each strand of the duplex
has a distinct nucleotide sequence, wherein only one nucleotide
sequence region (guide sequence or the antisense sequence) has
complementarity to a target nucleic acid sequence and the other
strand (sense sequence) comprises nucleotide sequence that is
homologous to the target nucleic acid sequence.
[0427] MicroRNAs (miRNA) are single-stranded RNA molecules,
typically of about 21-23 nucleotides in length, which regulate gene
expression. miRNAs are encoded by genes that are transcribed from
DNA but not translated into protein (non-coding RNA); instead they
are processed from primary transcripts known as pri-miRNA to short
stem-loop structures called pre-miRNA and finally to functional
miRNA. Mature miRNA molecules are partially complementary to one or
more messenger RNA (mRNA) molecules, and their main function is to
downregulate gene expression.
[0428] Certain RNA inhibiting agents may be utilized to inhibit the
expression or translation of messenger RNA ("mRNA") that is
associated with a cancer phenotype. Examples of such agents
suitable for use herein include, but are not limited to, short
interfering RNA ("siRNA"), ribozymes, and antisense
oligonucleotides. Specific examples of RNA inhibiting agents
suitable for use herein include, but are not limited to, Cand5,
Sirna-027, fomivirsen, and angiozyme.
[0429] (xii) Small molecule enzymatic inhibitors
[0430] Certain small molecule therapeutic agents are able to target
the tyrosine kinase enzymatic activity or downstream signal
transduction signals of certain cell receptors such as epidermal
growth factor receptor ("EGFR") or vascular endothelial growth
factor receptor ("VEGFR"). Such targeting by small molecule
therapeutics can result in anti-cancer effects. Examples of such
agents suitable for use herein include, but are not limited to,
imatinib, gefitinib, erlotinib, lapatinib, canertinib, ZD6474,
sorafenib (BAY 43-9006), ERB-569, and their analogues and
derivatives.
[0431] (xiii) Biological response modifiers
[0432] Certain protein or small molecule agents can be used in
anti-cancer therapy through either direct anti-tumor effects or
through indirect effects. Examples of direct-acting agents suitable
for use herein include, but are not limited to, differentiating
reagents such as retinoids and retinoid derivatives.
Indirect-acting agents suitable for use herein include, but are not
limited to, agents that modify or enhance the immune or other
systems such as interferons, interleukins, hematopoietic growth
factors (e.g., erythropoietin), and antibodies (monoclonal and
polyclonal).
[0433] (xiv) Anti-metastatic agents
[0434] The process whereby cancer cells spread from the site of the
original tumor to other locations around the body is termed cancer
metastasis. Certain agents have anti-metastatic properties,
designed to inhibit the spread of cancer cells. Examples of such
agents suitable for use herein include, but are not limited to,
marimastat, bevacizumab, trastuzumab, rituximab, erlotinib,
MMI-166, GRN163L, hunter-killer peptides, tissue inhibitors of
metalloproteinases (TIMPs), their analogues, derivatives and
variants.
[0435] (xv) Chemopreventative agents
[0436] Certain pharmaceutical agents can be used to prevent initial
occurrences of cancer, or to prevent recurrence or metastasis.
Examples of chemopreventative agents suitable for use herein
include, but are not limited to, tamoxifen, raloxifene, tibolone,
bisphosphonate, ibandronate, estrogen receptor modulators,
aromatase inhibitors (letrozole, anastrozole), luteinizing
hormone-releasing hormone agonists, goserelin, vitamin A, retinal,
retinoic acid, fenretinide, 9-cis-retinoid acid, 13-cis-retinoid
acid, all-trans-retinoic acid, isotretinoin, tretinoid, vitamin B6,
vitamin B 12, vitamin C, vitamin D, vitamin E, cyclooxygenase
inhibitors, non-steroidal anti-inflammatory drugs (NSAIDs),
aspirin, ibuprofen, celecoxib, polyphenols, polyphenol E, green tea
extract, folic acid, glucaric acid, interferon-alpha, anethole
dithiolethione, zinc, pyridoxine, finasteride, doxazosin, selenium,
indole-3-carbinal, alpha-difluoromethylornithine, carotenoids,
beta-carotene, lycopene, antioxidants, coenzyme Q10, flavonoids,
quercetin, curcumin, catechins, epigallocatechin gallate,
N-acetylcysteine, indole-3-carbinol, inositol hexaphosphate,
isoflavones, glucanic acid, rosemary, soy, saw palmetto, and
calcium. Cancer vaccines are an additional example of
chemopreventative agents suitable for use in the present invention.
These can be created through immunizing a patient with all or part
of a cancer cell type that is targeted by the vaccination
process.
[0437] (xvi) Side-effect limiting agents
[0438] Treatment of macrophage related diseases such as cancer with
oxidative or immunomodulatory agents alone or in combination with
other therapeutic compounds such as antineoplastic agents may be
accompanied by administration of pharmaceutical agents that can
alleviate the side effects produced by the antineoplastic agents.
Such agents suitable for use herein include, but are not limited
to, anti-emetics, anti-mucositis agents, pain management agents,
infection control agents, and anti-anemia/anti-thrombocytopenia
agents. Examples of anti-emetics suitable for use herein include,
but are not limited to, 5-hydroxytryptamine 3 receptor antagonists,
metoclopramide, steroids, lorazepam, ondansetron, cannabinoids,
their analogues and derivatives. Examples of anti-mucositis agents
suitable for use herein include, but are not limited to, palifermin
(keratinocyte growth factor), glucagon-like peptide-2, teduglutide,
L-glutamine, amifostin, and fibroblast growth factor 20. Examples
of pain management agents suitable for use herein include, but are
not limited to, opioids, opiates, and non-steroidal
anti-inflammatory compounds. Examples of agents used for control of
infection suitable for use herein include, but are not limited to,
antibacterials such as aminoglycosides, penicillins,
cephalosporins, tetracyclines, clindamycin, lincomycin, macrolides,
vancomycin, carbapenems, monobactams, fluoroquinolones,
sulfonamides, nitrofurantoins, their analogues and derivatives.
Examples of agents that can treat anemia or thrombocytopenia
associated with chemotherapy suitable for use herein include, but
are not limited to, erythropoietin and thrombopoietin.
[0439] XXIII. Neurological Diseases
[0440] In one embodiment, the present invention provides a method
of a neurological disease, such as amyotrophic lateral sclerosis
(ALS), Parkinson's disease and Alzheimer's disease, comprising
administering to a subject in need thereof an effective amount of a
chlorite-containing agent and optionally another therapy for
treating the disease. In some embodiments, the chlorite containing
agent is TCDO. In one example, the chlorite formulation is
WF10.
[0441] In one embodiment, the present invention provides a method
of treatment of neurological disease characterized by
neuroinflammation. Such diseases may include amyotrophic lateral
sclerosis (ALS), Parkinson's disease and Alzheimer's disease.
Multiple sclerosis (MS) may be excluded. In one embodiment, agents
capable of blocking migration as discussed herein such as
natalizumab (Tysabri.RTM.), fingolimod, or cladribine are used for
the treatment of non-MS neuroinflammation. Agents blocking
epithelial adhesion and/or chemotaxis as discussed herein are also
provided for treatment of non-MS neuroinflammation.
[0442] (a) Amyotrophic Lateral Sclerosis (ALS)
[0443] In some embodiments, the present invention provides methods
for treatment of amyotrophic lateral sclerosis (ALS) comprising
administering to a subject in need thereof an effective amount of
an oxidative agent (for example, chlorite or a chlorite-containing
agent, chloramine-T and hydrates thereof, and/or
1,3-dichloro-5,5-dimethylhydantoin) alone or in combination with
another agent used for treating the disease. The complications
related to ALS that can be treated with the methods of the present
invention include but are not limited to breathing problems, pain,
respiratory failure, eating disorders, malnutrition, dehydration,
choking, pneumonia, and dementia such as frontotemporal
dementia.
[0444] Amyotrophic lateral sclerosis is a form of motor neuron
disease. ALS, commonly referred to as Lou Gehrig's Disease, or
sometimes as Maladie de Charcot, is a progressive, fatal,
neurodegenerative disease caused by the degeneration of motor
neurons, the nerve cells in the central nervous system that control
voluntary muscle movement. The disorder causes muscle weakness and
atrophy throughout the body neurodegeneration of the motor neurons
in the brain (upper motor neurons) and spinal cord (lower motor
neurons). As a result, the neurons cease sending messages to
muscles. Unable to function, the muscles gradually weaken, develop
fasciculations because of denervation, and eventually atrophy. The
patient may ultimately lose the ability to initiate and control all
voluntary movement. They can also have difficulty breathing.
However, bladder and bowel sphincters and the muscles responsible
for eye movement are usually spared. Cognitive function is
generally unaffected by ALS except in certain situations such as
when ALS is associated with frontotemporal dementia (Phukan J,
Pender N P, Hardiman O 2007 Lancet Neurol 6 (11): 994-1003).
Sensory nerves and the autonomic nervous system, which controls
functions such as sweating, generally remain functional.
[0445] ALS affects people worldwide of all ages, ethnicity and
social classes. The onset of ALS has been linked to several
factors, including: a virus; exposure to neurotoxins or heavy
metals; DNA defects; immune system abnormalities; occupational
factors such as military service and elite sports; and enzyme
abnormalities. Surgeries involving the spinal cord have also been
thought to play a role in the onset of ALS due to the disruption of
nerve fibers. There is a known hereditary factor in familial ALS
(FALS); however, there is no known hereditary component in the
90-95% cases diagnosed as sporadic ALS. An inherited genetic defect
on chromosome 21 is associated with approximately 20% of familial
cases of ALS (Conwit, Robin A. (December 2006). Journal of the
Neurological Sciences 251 (1-2): 1-2; Al-Chalabi, Ammar; P. Nigel
Leigh (August 2000). Current Opinion in Neurology 13 (4): 397-405).
This mutation is believed to be autosomal dominant The children of
those diagnosed with familial ALS have a higher risk factor for
developing the disease; however, those who have close family
members diagnosed with sporadic ALS have no greater a risk factor
than the general population. Dietary intake of polyunsaturated
fatty acids (PUFA) has been shown in several studies to decrease
the risk of developing ALS (Veldink J H, et al. April 2007, J.
Neurol. Neurosurg. Psychiatr. 78 (4): 367-71).
[0446] The cause of ALS is not known, however, mutations in the
gene that produces the Cu/Zn superoxide dismutase (SOD1) enzyme are
associated with some cases (approximately 20%) of familial ALS.
This enzyme is an antioxidant that protects the body from damage
caused by superoxide, a toxic free radical. Free radicals can
accumulate and cause damage to DNA and proteins within cells.
Studies involving transgenic mice have yielded several theories
about the role of SOD1 in mutant SOD1 familial amyotrophic lateral
sclerosis. Mice lacking the SOD1 gene entirely do not customarily
develop ALS, although they do exhibit an acceleration of
age-related muscle atrophy (sarcopenia) and a shortened lifespan.
This indicates that the toxic properties of the mutant SOD1 are a
result of a gain in function rather than a loss of normal function.
In addition, aggregation of proteins has been found to be a common
pathological feature of both familial and sporadic ALS. In mutant
SOD1 mice, aggregates (misfolded protein accumulations) of mutant
SOD1 were found only in diseased tissues, and greater amounts were
detected during motor neuron degeneration (Furukawa Y, et al. 2006
Proc Natl Acad Sci USA 103 (18): 7148-53). It is speculated that
aggregate accumulation of mutant SOD1 plays a role in disrupting
cellular functions by damaging mitochondria, proteasomes, protein
folding chaperones, or other proteins (Boillee S, et al. 2006
Neuron 52 (1): 39-59). Other factors that may be associated with
ALS include but are not limited to glutamate.
[0447] There is no cure for ALS. Riluzole is currently the only
U.S. Food and Drug Administration (FDA) approved drug for ALS and
targets glutamate transporters. Riluzole is marketed by
Sanofi-Aventis S.A. with the brand name Rilutek.RTM.. Riluzole is
believed to reduce damage to motor neurons by decreasing the
release of glutamate via activation of glutamate transporters. In
addition, the drug offers a wide array of other neuroprotective
effects, by means of sodium and calcium channel blockades (Hubert J
P, Delumeau J C, Glowinski J, Premont J, Doble A. (1994). Br. J.
Pharmacol. 113 (1): 261-267), inhibition of protein kinase C, and
the promotion of NMDA (N-methyl d-aspartate) receptor antagonism
(Noh K M, Hwang J Y, Shin H C, Koh J Y. (2000). A Novel
Neuroprotective Mechanism of Riluzole: Direct Inhibition of Protein
Kinase C. Neurobiol Dis. 7 (4): 375-383; Beal M F, Lang A E,
Ludolph A C. (2005). Neurodegenerative Diseases: Neurobiology,
Pathogenesis and Therapeutics. Cambridge: Cambridge University
Press. p. p. 775). Clinical trials with ALS patients showed that
riluzole lengthens survival by several months, and may have a
greater survival benefit for those with a bulbar onset. The drug
also extends the time before a patient needs ventilation support.
Riluzole does not reverse the damage already done to motor neurons,
and patients taking the drug must be monitored for liver damage and
other possible side effects.
[0448] A small, open-label study recently suggested that the drug
lithium which traditionally is used for the treatment of bipolar
affective disorder may slow progression in both animal models and
the human form of ALS (Fornai F, Longone P, Cafaro L, et al.
(2008). Proc. Natl. Acad. Sci. U.S.A. 105: 2052).
[0449] The antibiotic ceftriaxone has demonstrated an unexpected
effect on glutamate and appears to be a beneficial treatment for
ALS in animal models. Ceftriaxone sodium is marketed by Hoffinan-La
Roche under the trade name Rocephin.RTM.. Ceftriaxone is currently
being tested in clinical trials.
[0450] Other drugs are undergoing development for ALS. For example,
KNS-760704 is under clinical investigation in ALS patients. It is
the enantiomer of pramipexole, which is approved for the treatment
of Parkinson's disease and restless legs syndrome (Abramova N A et
al. J Neurosci Res. 2002 Feb. 15; 67(4):494-500). However,
KNS-760704, which has been manufactured to a high degree of
enantiomeric purity and which is essentially inactive at dopamine
receptors, is not dose limited by the potent dopaminergic properies
of pramipexole (Gribkoff V and Bozik M. CNS Neurosci Ther. 2008
Fall; 14(3):215-26). The potential utility of KNS-760704 in ALS is
being advanced in clinical studies by Knopp Neurosciences Inc. of
Pittsburgh, Pa. The tetracycline antibiotic minocycline is also
under investigation for the treatment of ALS among other
neurological disorders. RNAi has been used in lab rats to shut off
specific genes that lead to ALS. RNAi gene silencing technology has
been used to target the mutant SOD1 gene (Xia X, Zhou H, Huang Y,
Xu Z (September 2006). Neurobiol Dis. 23 (3): 578-86). The mutant
SOD1 gene is responsible for causing ALS in a subset of the 10% of
all ALS patients who suffer from the familial, or genetic, form of
the disease.
[0451] The orally-administered drug arimoclomol is currently in
clinical evaluation as a therapeutic treatment for ALS. Arimoclomol
has been shown to extend life in an animal model of ALS. Kalmar B,
et al. Late stage treatment with arimoclomol delays disease
progression and prevents protein aggregation in the SOD1 mouse
model of ALS. J. Neurochem. 107:339-50 (2008).
[0452] Insulin-like growth factor 1 has also been studied as
treatment for ALS. IPLEX, which is a recombinant IGF-1 with Binding
Protein 3(IGF1BP3), has been issued to be used in a clinical trial
for ALS patients in Italy. Furthermore, methylcobalamin is being
studied in Japan and preliminary results show it significantly
lengthens survival time of ALS patients (Izumi Y, Kaji R (October
2007). Brain Nerve 59 (10): 1141-7).
[0453] Other treatments for ALS are designed to relieve symptoms
and improve the quality of life for patients. Physicians can
prescribe medications to help reduce fatigue, ease muscle cramps,
control spasticity, and reduce excess saliva and phlegm. Drugs also
are available to help patients with pain, depression, sleep
disturbances, dysphagia, and constipation. Physical therapy and
special equipment such as assistive technology can enhance patient
independence and safety throughout the course of ALS. Gentle,
low-impact aerobic exercise such as walking, swimming, and
stationary bicycling can strengthen unaffected muscles, improve
cardiovascular health, and help patients fight fatigue and
depression. Range of motion and stretching exercises can help
prevent painful spasticity and shortening (contracture) of muscles.
ALS patients who have difficulty speaking may benefit from working
with a speech-language pathologist. As ALS progresses,
speech-language pathologists can recommend the use of augmentative
and alternative communication such as voice amplifiers,
speech-generating devices (or voice output communication devices)
and/or low tech communication techniques such as alphabet boards or
yes/no signals. These methods and devices help patients communicate
when they can no longer speak or produce vocal sounds. When the
muscles that assist in breathing weaken, use of nocturnal
ventilatory assistance (intermittent positive pressure ventilation
(IPPV) or bilevel positive airway pressure (BIPAP)) may be used to
aid breathing during sleep. Such devices artificially inflate the
patient's lungs from various external sources that are applied
directly to the face or body. When muscles are no longer able to
maintain oxygen and carbon dioxide levels, these devices may be
used full-time. Patients may eventually consider forms of
mechanical ventilation (respirators) in which a machine inflates
and deflates the lungs. To be effective, this may require a tube
that passes from the nose or mouth to the trachea and for long-term
use, an operation such as a tracheostomy, in which a plastic
breathing tube is inserted directly in the patient's windpipe
through an opening in the neck. In addition, both animal and human
research suggest calorie restriction (CR) may be contraindicated
for those with ALS. Research on a transgenic mouse model of ALS
demonstrates that CR may hasten the onset of death in ALS (Hamadeh
M J, et al. (February 2005). Muscle Nerve 31 (2): 214-20). It has
also been found that in the ALS mouse model, CR "accelerates the
clinical course" of the disease and had no benefits (Pedersen W A,
Mattson M P (June 1999). Brain Res. 833 (1): 117-20), suggesting
that a calorically dense diet may slow ALS, a ketogenic diet in the
ALS mouse model has been shown to slow the progress of disease
(Zhao Z, Lange D J, Voustianiouk A, et al. (2006). BMC Neurosci 7:
29).
[0454] Potential therapeutic agents for ALS compounds include
immune modulators, complement and major histocompatibility complex
(MHC) inhibitors, protective agents for myelination and synaptic
connections, cellular stress agents, inhibitors of protein
expression, and neurotrophins. The present invention provides
methods of enhancing the efficacy of such agents by treating a
patient in need thereof with the agent in combination with chlorite
or a chlorite-containing agent.
[0455] In some embodiments, chlorite or a chlorite-containing
agents of the present invention are used in combination with immune
modulators for treatment of ALS. The immune response is believed to
play a role in the pathogenesis of ALS. In some cases, immune
factors may mitigate disease but in other cases exacerbate the
disease. Immune modulatory agents that provide treatment can be
used in combination with chlorite or a chlorite-containing agent.
The complement cascade may be upregulated in ALS patients. In some
embodiments, modulators of the complement cascade and major
histocompatibility complex (MHC) inhibitors can be used for
treatment of ALS in combination with chlorite or a
chlorite-containing agent.
[0456] In some embodiments, chlorite or a chlorite-containing agent
of the present invention are used in combination with protective
agents for myelination and synaptic connections for treatment of
ALS. Motor unit dysfunction in ALS may begin at the axon and at
synaptic connections. Agents and interventions that protect the
myelin sheath, remyelination, and synaptic remodeling can have
beneficial effects in ALS patients.
[0457] In some embodiments, chlorite or a chlorite-containing agent
of the present invention are used in combination with cellular
stress agents for treatment of ALS. Motor neurons in ALS may be
less capable of responding to stress caused by over-activity,
oxidative damage, misfolded proteins or other insults, when
compared to normal cells. Therapeutic agents that reduce stress or
bolster the vulnerable cells' natural stress response mechanisms
can have beneficial effect in ALS patients.
[0458] In some embodiments, chlorite or a chlorite-containing agent
of the present invention are used in combination with inhibitors of
protein expression for treatment of ALS. For example, inhibitors of
expression of genes that are known or suspected to produce or
exacerbate the ALS disease state can be used to treat ALS. In a
non-limiting example, RNAi can be used to downregulate the
expression of ALS genes including mutant SOD and TDP-43.
[0459] Chlorite or a chlorite-containing oxidative agent disclosed
herein can be used in combination with agents that are associated
with nerve regrowth for treatment of ALS. Factors that promote the
survival of neurons are referred to as neurotrophic factors.
Neurotrophic factors are secreted by target tissue and act by
preventing the associated neuron from initiating programmed cell
death, and induce differentiation of progenitor cells to form
neurons. Neurotrophins are a family of secreted proteins that
induce the survival, development and function of neurons.
Neurotrophins include nerve growth factor (NGF), brain-derived
neurotrophic factor (BDNF), neurotrophin-3 (NT-3), neurotrophin-4
(NT-4) and novel neurotrophin-1 (NNT1). Insulin-like growth
factor-1 (IGF1) acts on a wide variety of cell types to regulate
cell growth and development, especially in nerve cells. In some
embodiments of the present invention, chlorite or a
chlorite-containing agent of the present invention are used in
combination with neurotrophic factors and other factors that
stimulate nerve cell survival and differentiation to treat ALS.
Such factors can be delivered in various ways, e.g., via viral or
gene therapy or intraperitoneal injection. In some embodiments of
the present invention, chlorite or a chlorite-containing agent of
the present invention are used in combination with agents that
stimulate the activity or production of neurotrophic factors and
other factors that stimulate nerve cell survival and
differentiation to treat ALS.
[0460] In some embodiments, chlorite or a chlorite-containing agent
of the present invention are used in combination with molecules
that affect neurotransmitter levels for treatment of ALS. Examples
of such molecules include but are not limited to
5-hydroxytryptophan (5-HTP), tyrosine, levodopa (L-DOPA),
tryptophan, and cysteine.
[0461] Potential therapeutic agents for ALS are commonly tested in
ALS animal models, including the human Cu/Zn superoxide dismutase
(hSOD1) G93A transgene ALS mouse model (hSOD1-G93A mouse), which
exhibits high velocity of disease progression and short lifespan. A
comprehensive archive of ALS animal studies is available at the web
site of the ALS Therapy Development Institute (ALSTDI). In some
embodiments, chlorite or chlorite-containing agents are used to
treat ALS patients in combination with therapeutic agents or
interventions that have been tested in animal models, including but
not limited to 2-PMPA, adenosine, anisomycin, apocynin,
apomorphine, arimoclomol, aspergillin, BMP-7, carboxyfullerenes,
ceftriaxone, celastrol, geldanamycin, celecoxib (Celebrex.RTM.),
cyclooxygenase 2, CGP 3466B, chlorpromazine, clioquinol, clozapine,
ciliary neurotrophic factor (CTNF), colchicine, colivelin,
copaxone, copper chelators, lipoic acid, coenzyme Q10 (CoQ10),
creatine, curcumin, cytotoxic T-lymphocyte antigen 4 antibody
fusions (CTLA4-Ig), cycloheximide, cobra venom factor (CVF),
cycloserine, cyclosporin, d-penicillamine, JAK3/Dapsone,
Dapsone/Gusperimus/JAK3 Cocktail, Diethyldithiocarbamate DDC,
desferoxamine, desipramine, .alpha.-difluoromethylornithine (DFMO),
dietary restriction, dihydrotestosterone, 5,5-dimethyl-pyrroline
N-oxide (DMPO), excitatory amino acid transporter 2 (EAAT2),
erythro-9-[3-(2-hydroxynonyl)] adenine (EHNA), emetine, estradiol
benzoate, exercise, FK-506, fluorouracil, glial cell line-derived
neurotrophic factor (GDNF), decreased spinal copper levels,
genistein, glutamate receptor 3 (GLUR3) antisense, hepatocyte
growth factor (HGF), hNT neurons, anti-oxidant SOD1 protein, human
umbilical cord blood mononuclear cells, hydroxyurea,
interleukin-1beta-converting enzyme (ICE) inhibition, IGF-1 or
isoforms thereof, intravenous immunoglobulin (IVIG), indomethacin,
hydroquinone hydrochloride derivative of 17-AAG (IPI-504), iron
porphyrin, ivermectin (22,23-dihydroavermectin
B1a+22,23-dihydroavermectin B1b), L-acetyl-carnitine, lactacystin,
leflunomide, lentiviral RNAi SOD1 gene silencing, leukemia
inhibitory factor (LIF), lithium, lyophilized red wine extract,
magnesium supplementation, melatonin, memantine, metalloporphyrins
(MnTE-Py-P (AEOL10113 and AEOL10150)), metallothioneins, metformin,
methotrexate, mechano growth factor (MGF; IGF-I Ec peptide; mIGF-1
isoform), minocycline, minocycline/creatine,
minocycline/riluzole/nimodipine cocktail, mithramycin,
1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and/or
3-nitropropionic acid (3NP), N-acetyl-L-cysteine, N-acetylcysteine,
2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo[f]quinoxaline-2,3-dione
(NBQX), nordihydroguaiaretic acid (NDGA), neurofilament heavy
(NF-H) protein, neurofilament light (NF-L) protein, nimesulide,
nitric oxide synthase inhibitors, 17 beta-estradiol, p75
neurotrophin receptor, p75 neurotrophin agonist, p75 neurotrophin
antisense, parvalbumin, sodium phenyl butyrate (PBA), peripheral
axotomy, phosphatidyl choline-bound Cu/Zn SOD, pioglitazone,
polyamine-modified catalase, porphyrin, prednisolone, progesterone,
puromycin, putrescine-modified catalase (PUT-CAT), quinacrine, R(+)
pramipexole, radicicol, rasagiline, resveratrol/red wine extract,
riluzole, ritonavir, anti-myostatin mAb, vascular endothelial
growth factor (VEGF), RNAi targeting human SOD1 gene, rofecoxib,
rolipram, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid
antagonist RPR 119990, 5-hydroxytryptophan (5-HTP), sodium
valproate, stem cells, sulindac, tamoxifen, propargylamine TCH346,
tepoxalin, testosterone, thalidomide, trehalose, trichostatin A,
trientine/ascorbate, vincristine, vitamin E/riluzole/gabapentin,
Janus kinase 3 (JAK3) inhibitor such as WHI-P131, bone marrow
transplant, zileuton, zinc sulfate, or noncompetitive AMPA
antagonist such as ZK 187638. In some embodiments, these agents
have a beneficial effect in animal models of ALS and human ALS
patients. In some embodiments, these agents have a beneficial
effect in animal models of ALS but have attenuated or no effect in
human ALS patients. Indeed, there are many therapeutic agents that
show activity in animal models but have reduced activity, or no
beneficial activity, when administered to human patients. Without
being bound by any theory, it is believed that therapeutic agents
that address neuronal function (e.g., riluzole and agents that act
on glutamate receptors or facilitate mitochondria repair) and
neuronal regrowth (e.g., IGF1) would be more effective if the
diseased inflammatory environment is at least partially resolved.
Thus, many of the ALS treatments that show activity in animal
models (e.g., the hSOD1-G93A mouse) but have failed in humans will
be more effective for treating ALS after the inflammatory
environment has been treated, e.g., in combination with chlorite or
a chlorite-containing agent of the present invention. Thus,
chlorite and chlorite-containing agents can be used to "rescue"
drugs and other therapies or interventions.
[0462] Similarly, some of the above agents or interventions have
shown a beneficial effect during in vitro studies but have
attenuated or no effect in animal models of ALS or in human ALS
patients. In some embodiments, such agents are efficacious in vivo
when used in combination with chlorite or a chlorite-containing
agent.
[0463] The methods of the invention provide administering chlorite
or a chlorite-containing agent in combination with one or more
other therapeutic agents or interventions that are being developed
to treat ALS or are promising for treating ALS. In some
embodiments, chlorite or a chlorite-containing agent is
administered in combination with therapeutic agents during
preclinical and/or clinical development of such agents. In some
embodiments, these therapeutic agents have shown therapeutic
benefit during in vitro or in vivo studies. In some embodiments,
these agents are known to treat other diseases, and have an
activity or benefit that is suspected to be useful in the ALS
setting. In a non-limiting example, neuroprotective or
anti-inflammatory agents, e.g., agents known to be useful for other
diseases or disorders, may be suspected to also have a beneficial
effect in ALS patients. Such promising agents can be administered
in combination with chlorite or a chlorite-containing agent to
further enhance their beneficial effect. A number of potential
points of therapeutic intervention have been identified for
treatment of ALS. ALS targets include those related to apoptosis,
axon growth/transport, cell cycle, cell replacement,
excitotoxicity, functional intervention, inflammation, oxidative
stress and transcription. Targets further include trophic factors,
biomarkers, proteins, mitochondria, and muscle factors. In some
embodiments, the invention provides methods to treat ALS in a
subject in need thereof comprising administration of chlorite or a
chlorite-containing agent in combination with another agent that
acts upon one or more ALS targets.
[0464] In some embodiments, the target for ALS treatment is a 5-HT
receptor. Non-limiting examples of target related treatments
include Buspirone. In some embodiments, the target for ALS
treatment is a 5-LOX Inhibitor. Non-limiting examples of target
related treatments include GPI-20359. In some embodiments, the
target for ALS treatment is an ACE inhibitor. Non-limiting examples
of target related treatments include Ramipril and Teveten. In some
embodiments, the target for ALS treatment is an ADA inhibitor.
Non-limiting examples of target related treatments include EHNA. In
some embodiments, the target for ALS treatment is an Adenosine
receptor. Non-limiting examples of target related treatments
include CGS21680. In some embodiments, the target for ALS treatment
is an adhesion molecule. Non-limiting examples of target related
treatments include natalizumab (Tysabri.RTM.), Efomycine M,
LGD5552, Lovenox or purified phenolic glycolipid (PGL). In some
embodiments, the target for ALS treatment is an Adjunct Therapy.
Non-limiting examples of target related treatments include
BMS-387032, Cereport or Nicotine. In some embodiments, the target
for ALS treatment is an Aggregate or Prion. Non-limiting examples
of target related treatments include
2-amino-4,7-dimethyl-benzothiazol-6-ol, 2-methoxyestradiol, Apan,
Chlorpromazine, CPHPC, Cyclodextrin, Fibrillex, Guanidine
hydrochloride, Heparin, hSOD1, MG-132, Piroxicam, Porphyrin,
Promethazine, Protoporphyrin, Y-27632, or Melatonin. In some
embodiments, the target for ALS treatment is an AIDS drug.
Non-limiting examples of target related treatments include
Indinavir, or Kaletra.RTM. (ritonavir+lopinavir). In some
embodiments, the target for ALS treatment is an AKT Inhibitor.
Non-limiting examples of target related treatments include KP372-1,
LY294002, Perifosine, Tricirbine, or Wortmannin. In some
embodiments, the target for ALS treatment is an Alternate cellular
energy source. Non-limiting examples of target related treatments
include ATP or Trehalose. In some embodiments, the target for ALS
treatment is an AMPA Receptor. Non-limiting examples of target
related treatments include GLUR3 antisense, GYKI-47261, NBQX, RPR
119990, RPR117824, Temocapril, ZK 187638, or Ivermectin. In some
embodiments, the target for ALS treatment is an Anti-FAS.
Non-limiting examples of target related treatments include
Antisense vs FAS, MFL3 or R-125224. In some embodiments, the target
for ALS treatment is an Anti-Microtubule. Non-limiting examples of
target related treatments include Liposome-Based Taxol or Taxol. In
some embodiments, the target for ALS treatment is an
anticholinesterase. Non-limiting examples of target related
treatments include neostigmine or physostigmine. In some
embodiments, the target for ALS treatment is an Anticoagulant.
Non-limiting examples of target related treatments include
Desmoteplase or Enoxaparin. In some embodiments, the target for ALS
treatment is an antioxidant. Non-limiting examples of target
related treatments include glutathione, Glutathione peroxidase,
N-acetyl-L-cysteine, procysteine, PUT-CAT, TR500, Underexpression
of MnSOD, Melatonin or hSOD1. In some embodiments, the target for
ALS treatment is an Antiviral agent. Non-limiting examples of
target related treatments include IFN-beta, Pleconaril or tilorone.
In some embodiments, the target for ALS treatment is apoptosis.
Non-limiting examples of target related treatments include Bc1-2.
In some embodiments, the target for ALS treatment is
Astrocyte/Microglial replacement. Non-limiting examples of target
related treatments include Cord Blood cells, Cord Blood ICV or
Diflunisal. In some embodiments, the target for ALS treatment is an
Astrocytes/Microglia. Non-limiting examples of target related
treatments include Alpha GPE, Aminoadipic acid, Anti-PS receptor
neutralizing antibody, Campath, IL-4PE38, Interferon gamma,
Melatonin, Mycophenolate Mofetil, RI273, Tetrathiomolybdate, tissue
plasminogen activator antisense, Tranilast or Neuroserpin. In some
embodiments, the target for ALS treatment is Autophagy.
Non-limiting examples of target related treatments include
3-methyladenine or Rapamycin. In some embodiments, the target for
ALS treatment is Axonal regeneration. Non-limiting examples of
target related treatments include Chondroitinase ABC, NEP1-40,
NgR(310)ecto-Fc or inosine. In some embodiments, the target for ALS
treatment is Axonal transport. Non-limiting examples of target
related treatments include Brimonidine or EHNA. In some
embodiments, the target for ALS treatment is a Bax Inhibitor.
Non-limiting examples of target related treatments include Bax
antisense, BIP, hTERT, Humanin, XIAP or colivelin. In some
embodiments, the target for ALS treatment is a Bc1 inducer.
Non-limiting examples of target related treatments include
ginsenoside Rb1 and Rg1, or Genasense. In some embodiments, the
target for ALS treatment is a calcium channel blocker. Non-limiting
examples of target related treatments include nimodipine or
verapamil. In some embodiments, the target for ALS treatment is a
Calpain Inhibitor. Non-limiting examples of target related
treatments include E64, Neurodur, PD150606, SJA6017 or calpeptin.
In some embodiments, the target for ALS treatment is a Caspase
Inhibitor. Non-limiting examples of target related treatments
include ESPA-1002, IDN-6556, M826, Q-VD-OPH, zVAD-FMK or variants
thereof or VX-740. In some embodiments, the target for ALS
treatment is a CDK inhibitor. Non-limiting examples of target
related treatments include Alsterpaullone, aragusterol A, AT7519,
AT9311, Butryrolactone, CDC25 Inhibitor, CGP60474, CINK-4, CYC202,
E7389, fascaplysin, Indirubin, Ken Kosik compounds 2 or 3,
kenpaullone, KN-93, Olomoucine, PD 0183812, Roscovitine,
indolo[6,7-a]pyrrolo[3,4-c]carbazoles, SQ-67563, Staurosporine,
TNP-470 or UCN-01. In some embodiments, the target for ALS
treatment is Cell Adhesion. Non-limiting examples of target related
treatments include KDI tripeptide. In some embodiments, the target
for ALS treatment is Cell Therapy. Non-limiting examples of target
related treatments include Immune modulation therapy, Neural stem
cells, Stem cells in ALS Rat or BM-NSC. In some embodiments, the
target for ALS treatment is a Ceramide. Non-limiting examples of
target related treatments include Cycloserine, D-Cycloserine or
ISP-1. In some embodiments, the target for ALS treatment is a
Chaperone Inducer or Co-inducer. Non-limiting examples of target
related treatments include celastrol, curcumin, radicicol. In some
embodiments, the target for ALS treatment is a chelator.
Non-limiting examples of target related treatments include
clioquinol, D-penicillamine or trientine/ascorbate. In some
embodiments, the target for ALS treatment is a Chemokine receptor
antagonist. Non-limiting examples of target related treatments
include AMD-3100, BX471, MLN1202, SCH-D, TAK-779 or vMIPII. In some
embodiments, the target for ALS treatment is a Complement
inhibitor. Non-limiting examples of target related treatments
include CVF. In some embodiments, the target for ALS treatment is a
copper chelator. Non-limiting examples of target related treatments
include penicillamine. In some embodiments, the target for ALS
treatment is Corticosteroid. Non-limiting examples of target
related treatments include Adrenalectomy. In some embodiments, the
target for ALS treatment is a COX inhibitor. Non-limiting examples
of target related treatments include sulindac. In some embodiments,
the target for ALS treatment is a selective COX-2 Inhibitor.
Non-limiting examples of target related treatments include
valdecoxib (Bextra.RTM.), celecoxib (Celebrex.RTM.), nimesulide,
rofecoxib or SC-236. In some embodiments, the target for ALS
treatment is a COX/LOX inhibitor. Non-limiting examples of target
related treatments include licofelone or tepoxalin. In some
embodiments, the target for ALS treatment is Cross Linking.
Non-limiting examples of target related treatments include ALT-711.
In some embodiments, the target for ALS treatment is a CXCR3
Antagonist. In some embodiments, the target for ALS treatment is a
Cysteine Protease Inhibitor. Non-limiting examples of target
related treatments include CA-074, CRA-3316 or Cystatin C. In some
embodiments, the target for ALS treatment is a Cytochrome Enyme
Inducer. Non-limiting examples of target related treatments include
Isoniazid. In some embodiments, the target for ALS treatment is a
Cytokine. In some embodiments, the target for ALS treatment is a
Cytokine Inhibitor. Non-limiting examples of target related
treatments include CP-SOCS3, etanercept (Enbrel.RTM.), adalimumab
(Humira.RTM.), infliximab (Remicade.RTM.), LMP-420 or naltrexone.
In some embodiments, the target for ALS treatment is a DNA
intercalator. Non-limiting examples of target related treatments
include Cyclophosphamide, Mithramycin, Mitomycin C or Quinacrine.
In some embodiments, the target for ALS treatment is DNA synthesis.
Non-limiting examples of target related treatments include MB7133,
Procarbazine, Raltitrexed or Leucovorin. In some embodiments, the
target for ALS treatment is Dopamine. Non-limiting examples of
target related treatments include Apomorphine. In some embodiments,
the target for ALS treatment is a dopamine agonist. Non-limiting
examples of target related treatments include bromocriptine. In
some embodiments, the target for ALS treatment is E2F-1/Sp1.
Non-limiting examples of target related treatments include E2F
Decoy, microgonotropen, Mitoxantrone Liposome or
Tallimustine-Distamycin. In some embodiments, the target for ALS
treatment is EAAT2. Non-limiting examples of target related
treatments include Ceftriaxone, Sodium Valproate or ONO-2506. In
some embodiments, the target for ALS treatment is ER stress.
Non-limiting examples of target related treatments include
salubrinal or IPI-504. In some embodiments, the target for ALS
treatment is Estrogen. Non-limiting examples of target related
treatments include estrone quinol, Genistein, raloxifene or
Tamoxifen. In some embodiments, the target for ALS treatment is
excitotoxicity. Non-limiting examples of target related treatments
include dextromethorphan, L-threonine, lamotrigine, levodopa or
topiramate. In some embodiments, the target for ALS treatment is a
Farnesyl transferase (FTase) inhibitor. Non-limiting examples of
target related treatments include BMS-214662 or tipifarnib. In some
embodiments, the target for ALS treatment is a GAPDH Inhibitor.
Non-limiting examples of target related treatments include
Pentalenolactone or TCH-346. In some embodiments, the target for
ALS treatment is Gene Therapy. Non-limiting examples of target
related treatments include CT GalNAc transferase or rabies G (RabG)
protein pseudotyped lentiviral vector EIAV.LacZ. In some
embodiments, the target for ALS treatment is a glucocorticoid
receptor agonist or Dexamethasone. In some embodiments, the target
for ALS treatment is Glutamate release. Non-limiting examples of
target related treatments include Riluzole. In some embodiments,
the target for ALS treatment is a Growth Factor. Non-limiting
examples of target related treatments include Agrin, Aranesp, CEPO,
Epoetin Beta, Erythropoietin, HF0299, MGF, MMP-9 and stem cells,
oprelvekin (Neumega.RTM.), RI624, thrombospindin or RK35. In some
embodiments, the target for ALS treatment is a GSK3beta inhibitor.
In some embodiments, the target for ALS treatment is a HAT
inhibitor. Non-limiting examples of target related treatments
include garcinol. In some embodiments, the target for ALS treatment
is a HDAC activator. Non-limiting examples of target related
treatments include Resveratrol or other red wine extract. In some
embodiments, the target for ALS treatment is a HDAC inhibitor.
Non-limiting examples of target related treatments include FK228,
Flavin analog DPD, MNTF, MS-27-275, PBA, Pivanex, PXD101, SAHA
(Suberoylanilide hydroxamine), Scriptaid, Trichostatin A or Sodium
Valproate. In some embodiments, the target for ALS treatment is a
Heat Shock Protein. Non-limiting examples of target related
treatments include arimoclomol. In some embodiments, the target for
ALS treatment is a Heat Shock Protein Inhibitor/Activator.
Non-limiting examples of target related treatments include 17 DMAG
or IPI-504. In some embodiments, the target for ALS treatment is a
HIF-1 Inducer. Non-limiting examples of target related treatments
include Cobalt. In some embodiments, the target for ALS treatment
is a HIV Protease inhibitor. Non-limiting examples of target
related treatments include amprenavir, fosamprenavir, nelfinavir,
nelfinavir/ritonavir or saquinavir. In some embodiments, the target
for ALS treatment is Hypoxia. Non-limiting examples of target
related treatments include Blood Substitute. In some embodiments,
the target for ALS treatment is an IL-1 inhibition. In some
embodiments, the target for ALS treatment is Immune Regulation.
Non-limiting examples of target related treatments include
Alzhemed, Ambrotose, Anthrax lethal factor, Anti-IL15 Antibody,
Anti-TNF antibody, Brefeldin A, CAMPATH-1H, CD45 antibody,
Copaxone, Dapsone, Dapsone-Gusperimus-JAK3 Cocktail,
Dapsone-Gusperimus Combo, Dapsone ICV, Desipramine,
DFMO-Chlorpromazine, DFMO-JAK3-Ritonavir,
DFMO-Ritonavir-Chlorpromazine, DHM2EQ, Early Pregnancy Factor,
Fludarabine, GMDP, Granzyme inhibitor, IL-10, IL-10 Gene Therapy,
Imiquimod, Immunokine, inosiplex, interferon alpha, isoprinosine,
levamisole, Luteolin, Mac 1 SAP, Mycotoxins, NK 1.1, Omeprazole,
Prednisolone, rFAS-ligand, RS-1178, Squalestatin, Suramin, total
lymphoid irradiation, TP10 or sCR1, Cyclosporin, Lipitor or
methylprednisolone. In some embodiments, the target for ALS
treatment is inflammation. Non-limiting examples of target related
treatments include bovine brain gangliosides, gangliosides, VX 148
or Sanglifehrin A. In some embodiments, the target for ALS
treatment is an ion channel inhibitor. Non-limiting examples of
target related treatments include amantadine and guanidine. In some
embodiments, the target for ALS treatment is a JAK/STAT.
Non-limiting examples of target related treatments include
leflunomide or WHI-P131. In some embodiments, the target for ALS
treatment is a JNK Inhibitor. Non-limiting examples of target
related treatments include CC-105, Colostrinin, AS601245 or
SP600125. In some embodiments, the target for ALS treatment is a
Kinase Inhibitor. Non-limiting examples of target related
treatments include PKC412, GW5074 or Y-27632. In some embodiments,
the target for ALS treatment is kinase upregulation. Non-limiting
examples of target related treatments include forskolin. In some
embodiments, the target for ALS treatment is a PARP inhibitor.
Non-limiting examples of target related treatments include GPI15427
& 16539, inosine or PJ-34. In some embodiments, the target for
ALS treatment is a phosphodiesterase inhibitor. Non-limiting
examples of target related treatments include Ariflo, MEM1414,
Roflumilast, Rolipram, SelCID-3, Viagra or EHNA. In some
embodiments, the target for ALS treatment is a Polo-like Kinase
Inhibitor. Non-limiting examples of target related treatments
include HMN-214. In some embodiments, the target for ALS treatment
is a Polyamine analog. Non-limiting examples of target related
treatments include Tetramines. In some embodiments, the target for
ALS treatment is a Polyamine inhibitor. Non-limiting examples of
target related treatments include mitoguazone or Xyloside. In some
embodiments, the target for ALS treatment is a potassium channel
blocker. Non-limiting examples of target related treatments include
3,4-diaminopyridine. In some embodiments, the target for ALS
treatment is a Protease Inhibitor. In some embodiments, the target
for ALS treatment is a Proteasome. Non-limiting examples of target
related treatments include Aclacinomycin A, Alpha-methylomuralide,
Aspergillin, CBZ-GPFL-CHO, CVT-634, Dantrolene, Dihydroeponemycin,
Epoxomicin, ICV Ritonavir, IDE Antibody, Lactacystin, Lovastatin,
p27 E3, Rasagiline, Ritonavir, Simvastatin, Torbafylline or
Velcade. In some embodiments, the target for ALS treatment is a
Protein Kinase C inhibitor. Non-limiting examples of target related
treatments include ISSI-3521, LY-333531, PEP005 or Propofol. In
some embodiments, the target for ALS treatment is a Protein
Synthesis Inhibitor. Non-limiting examples of target related
treatments include Hygromycin B, linezolid (Zyvox.RTM.) or
anisomycin. In some embodiments, the target for ALS treatment is
Protein Transport. Non-limiting examples of target related
treatments include 17-beta-Estradiol. In some embodiments, the
target for ALS treatment is REDOX. Non-limiting examples of target
related treatments include AGIX-4207, Ebselen, pyrroloquinoline
quinone or Resveratrol/red wine extract. In some embodiments, the
target for ALS treatment is a Ribonucleotide reductase inhibitor.
Non-limiting examples of target related treatments include
Desferoxamine, Hydroxyurea or Trimidox. In some embodiments, the
target for ALS treatment is a RNA synthesis inhibitor. In some
embodiments, the target for ALS treatment is a Serine Protease. In
some embodiments, the target for ALS treatment is a
serine/threonine kinase inhibitor. Non-limiting examples of target
related treatments include VX 680. In some embodiments, the target
for ALS treatment is SOD1. Non-limiting examples of target related
treatments include DDC or Temozolamide. In some embodiments, the
target for ALS
treatment is a SOD1 Inhibitor. Non-limiting examples of target
related treatments include Dietary copper deficiency (with
Clioquinol), Estradiol Benzoate, Histidine or Norethindrone. In
some embodiments, the target for ALS treatment is a stem cell.
Non-limiting examples of target related treatments include Human
umbilical cord blood. In some embodiments, the target for ALS
treatment is a TACE inhibitor (TNF-a). Non-limiting examples of
target related treatments include TMI-1. In some embodiments, the
target for ALS treatment is a TGF beta inhibitor. Non-limiting
examples of target related treatments include AP12009, inhibitors
of E3 ubiquitin ligases or Pirfenidone. In some embodiments, the
target for ALS treatment is a TNF-Alpha. Non-limiting examples of
target related treatments include Bupropion, Gemfibrozil,
Pirfenidone, Sepsis Vaccine or Thalidomide. In some embodiments,
the target for ALS treatment is a Topoisomerase I inhibitor.
Non-limiting examples of target related treatments include ARQ501,
Liposome Encapsulated SN38, Rebeccamycin Derivative or SN38. In
some embodiments, the target for ALS treatment is a Trophic Factor.
Non-limiting examples of target related treatments include
AAV-GDNF, Basic fibroblast growth factor, BDNF, BMP-7,
cardiotrophin-1, CNTF, GDNF, adenovirus-mediated GDNF, GDNF AAV,
growth hormone, HGF, Lantus, leukemia inhibitory factor (LIF),
neurophilin ligands, NF-H or NF-L overexpression, Olmesartan,
p75NTR Antagonist, rHCNTF, Thymic, thyrotropin, TRH or Xaliproden
hydrochloride. In some embodiments, the target for ALS treatment is
a Tyrosine kinase inhibitor. Non-limiting examples of target
related treatments include GLEEVEC.RTM. (imatinib mesylate) or
IRESSA.RTM. (gefitinib). In some embodiments, the target for ALS
treatment is Ubiquitin inhibition. Non-limiting examples of target
related treatments include Leu-Ala or UCHL1. In some embodiments,
the target for ALS treatment is a miscellaneous target.
Non-limiting examples of related treatments include Cisplatin,
Deprenyl or Nelfinavir/Combivir. In some embodiments, the target
for ALS treatment is Zinc Finger Transcription Disruption.
Non-limiting examples of target related treatments include
DIBA.
[0465] In some embodiments, the present invention provides methods
for treatment of amyotrophic lateral sclerosis (ALS) comprising
administering to a subject in need thereof chlorite or a
chlorite-containing agent in combination with another therapy. In
some embodiments, the present invention provides methods for
treatment of amyotrophic lateral sclerosis (ALS) comprising
administering to a subject in need thereof chlorite or a
chlorite-containing agent in combination with any one or more of
the drugs disclosed hereinabove. In some embodiments, chlorite or a
chlorite containing agent of the present invention is used in
combination with riluzole. In some embodiments, chlorite or a
chlorite containing agent of the present invention is used in
combination with KNS-760704. In some embodiments, chlorite or a
chlorite containing agent of the present invention is used in
combination with minocycline. In some embodiments, chlorite or a
chlorite containing agent of the present invention is used in
combination with RNAi targeting SOD1 gene. In some embodiments,
chlorite or a chlorite containing agent of the present invention is
used in combination with insulin-like growth factor 1 for the
treatment of ALS.
[0466] (b) Parkinson's disease (PD)
[0467] In some embodiments, the present invention provides methods
for treatment of Parkinson's disease or Parkinson's-like disease
comprising administering to a subject in need thereof an effective
amount of chlorite or a chlorite-containing agent. The
complications related to Parkinson's disease that can be treated
with the methods of the present invention include but are not
limited to motor impairment, severe tremor, injury related to motor
impairment, pain, dysphagia, constipation, bladder control and
urinary incontinence, speech problems, depression, compulsive
behavior, cognitive impairment, dementia, muscle rigidity
(akinesia), hallucination, visual problems, sleep disorders,
restless legs syndrome, leg cramps, impaired sexuality, worsened
sense of smell, and osteoporosis.
[0468] Parkinson's disease (PD) is a progressive degenerative
disease of the central and peripheral nervous systems. The risk of
developing Parkinson's disease increases with age, and afflicted
individuals are usually adults over 40. Parkinson's disease occurs
in all parts of the world, and affects more than one million
individuals in the United States alone. There are several other
conditions that have the features of Parkinson's disease and are
referred to as Parkinson's-like diseases. Parkinson's and
Parkinson's-like diseases can be characterized by tremor,
hypokinesia, rigidity, and postural instability.
[0469] The underlying causes of Parkinson's disease and
Parkinson's-like diseases are numerous, and diagnosis can be
complex. Parkinson's disease or Parkinson's-like disease is
characterized by degeneration of dopaminergic neurons of the
substantia nigra. The substantia nigra is a portion of the lower
brain or brain stem that helps control voluntary movements. The
shortage of dopamine in the brain caused by the loss of these
neurons may cause the observable disease symptoms.
[0470] Parkinson's disease is a chronic disorder that requires
broad-based management including but not limited to patient and
family education, support group services, general wellness
maintenance, physiotherapy, exercise, and nutrition. Medications or
surgical intervention can provide relief from the symptoms.
Medications for Parkinson's disease include but are not limited to
levodopa (L-3,4-dihydroxyphenylalanine; L-DOPA), catechol-O-methyl
transferase (COMT) inhibitors, dopamine agonists, monoamine oxidase
B (MAO-B) inhibitors, surgery and deep brain stimulation, and
neurorehabilitation.
[0471] The most widely used form of treatment is L-DOPA in various
forms. L-DOPA is transformed into dopamine in the dopaminergic
neurons by L-aromatic amino acid decarboxylase (often known by its
former name dopa-decarboxylase). However, only 1-5% of L-DOPA
enters the dopaminergic neurons. The remaining L-DOPA is often
metabolised to dopamine elsewhere, causing a wide variety of side
effects. Due to feedback inhibition, L-DOPA results in a reduction
in the endogenous formation of L-DOPA, and so eventually becomes
counterproductive. Carbidopa and benserazide are dopa decarboxylase
inhibitors. They help to prevent the metabolism of L-DOPA before it
reaches the dopaminergic neurons and are generally given as
combination preparations of carbidopa/levodopa (co-careldopa)
(e.g., Sinemet.RTM., Parcopa.RTM.) and benserazide/levodopa
(co-beneldopa) (e.g., Madopar.RTM.). There are also controlled
release versions of Sinemet.RTM. and Madopar.RTM. that spread out
the effect of the L-DOPA. Duodopa.RTM. is a combination of levodopa
and carbidopa, dispersed as a viscous gel. Using a patient-operated
portable pump, the drug is continuously delivered via a tube
directly into the upper small intestine, where it is rapidly
absorbed. Medications combining carbidopa, levodopa and entacapone
are also used (Stalevo.RTM.). The use of amantadine
(Symmetrel.RTM.), benztropine (Cogentin.RTM.), procyclidine
(Kemadrin.RTM.), or trihexyphenidyl (Artane.RTM.) with
levodopa-carbidopa can enhance the anti-Parkinson's effects of
levodopa.
[0472] COMT inhibitors can be used for treating PD. Tolcapone
inhibits the COMT enzyme, thereby prolonging the effects of L-DOPA,
and so has been used to complement L-DOPA. However, due to its
possible side effects such as liver failure, it is limited in its
availability. A similar drug, entacapone has not been shown to
cause significant alterations of liver function and maintains
adequate inhibition of COMT over time (Gelb D, Oliver E, Gilman S
(1999) Arch Neurol 56 (1): 33-9).
[0473] The dopamine agonists bromocriptine, pergolide, pramipexole,
ropinirole, cabergoline, apomorphine, and lisuride are moderately
effective for treating PD. These have their own side effects
including those listed above in addition to somnolence,
hallucinations and/or insomnia. Several forms of dopamine agonism
have been linked with a markedly increased risk of problem
gambling. Dopamine agonists initially act by stimulating some of
the dopamine receptors. However, they cause the dopamine receptors
to become progressively less sensitive, thereby eventually
increasing the symptoms.
[0474] MAO-B inhibitors such as selegiline and rasagiline reduce
the symptoms by inhibiting monoamine oxidase-B (MAO-B), thereby
inhibiting the breakdown of dopamine secreted by the dopaminergic
neurons. Metabolites of selegiline include levoamphetamine and
levomethamphetamine. This might result in side effects such as
insomnia A potential side effect of selegiline in conjunction with
L-DOPA is stomatitis, an inflammation of the mucous lining of any
of the structures in the mouth.
[0475] In addition to medication, deep brain stimulation is
presently the most used surgical means of treatment, but other
surgical therapies that have shown promise include surgical lesion
of the subthalamic nucleus and of the internal segment of the
globus pallidus, a procedure known as pallidotomy (Guridi J, Obeso
J A (2001) Brain 124 (Pt 1): 5-19; Fukuda M, Kameyama S, Yoshino M,
Tanaka R, Narabayashi H (2000) Stereotactic and functional
neurosurgery 74 (1): 11-20). Regular physical exercise and/or
therapy can be beneficial to the patient for maintaining and
improving mobility, flexibility, strength, gait speed, and quality
of life; and speech therapy may improve voice and speech
function.
[0476] In some embodiments, chlorite or a chlorite-containing agent
of the present invention can be used in combination with levodopa,
COMT inhibitors, dopamine agonists, MAO-B inhibitors, surgery, deep
brain stimulation, or neurorehabilitation for the treatment of
Parkinson's disease or Parkinson's-like diseases.
[0477] In some embodiments, the methods of the present invention
comprise administration of chlorite or a chlorite-containing
compound in combination with other therapeutic agents and/or
interventions that are used for the treatment of Parkinson's
disease and related complications. In some embodiments, chlorite or
a chlorite-containing agent of the present invention can be used in
combination with a neuroprotective agent or therapy for treating
Parkinson's disease. The neuroprotective agent or therapy can be
exercise, antioxidants, immunosuppressive calcineurin inhibitors,
nitric oxide synthase (NOS) inhibitors, sigma-1 modulators, AMPA
antagonists, Ca.sup.2+ channel blockers, estrogen agonists, MAO-B
inhibitors, kinase inhibitors, mitochondrial modulators or
enhancers, alpha synuclein modulators, glycoprotein IIb/IIIa
antagonists, erythropoietin, astaxanthin, boswellia, caffeine,
curcumin, E vitamins, tocotrienols, flavonoids, naringenin,
huperzine, or ubiquinol. In one embodiment, chlorite or a
chlorite-containing agent of the present invention is used in
combination with levodopa. In another embodiment, chlorite or a
chlorite-containing agent of the present invention is used in
combination with a COMT inhibitor such as tolcapone and entacapone.
In another embodiment, chlorite or a chlorite-containing agent of
the present invention is used in combination with a dopamine
agonist including but not limited to bromocriptine, pergolide,
pramipexole, ropinirole, cabergoline, apomorphine, and lisuride. In
another embodiment, chlorite or a chlorite-containing agent of the
present invention is used in combination with a MAO inhibitor
including but not limited to selegiline and rasagiline. In yet
another embodiment, chlorite or a chlorite-containing agent of the
present invention is used in combination with deep brain
stimulation and/or neurorehabilitation such as physical exercise or
therapy.
[0478] (c) Alzheimer's Disease (AD)
[0479] In some embodiments, the present invention provides methods
for treatment of Alzheimer's disease comprising administering to a
subject in need thereof an effective amount of chlorite or a
chlorite-containing agent. The complications related to Alzheimer's
disease that can be treated with the methods of the present
invention include but are not limited to memory loss,
sleeplessness, agitation, disorganization, wandering, anxiety,
depression, cognition disintegration, personality disintegration,
cognitive impairment, infections, severe urinary tract infections,
slow surgical recovery, chronic brain failure, anosmia, Hirano
body, pneumonia, and injuries from falls and other motor
impairment.
[0480] Alzheimer's disease (AD), also called Alzheimer disease,
Senile Dementia of the Alzheimer Type (SDAT) or simply Alzheimer's,
is the most common form of dementia. Generally it is diagnosed in
people over 65 years of age, although the less-prevalent
early-onset Alzheimer's can occur much earlier. Although each
sufferer experiences Alzheimer's in a unique way, there are many
common symptoms. The earliest observable symptoms are often
mistakenly thought to be `age-related` concerns, or manifestations
of stress. In the early stages, the most commonly recognized
symptom is memory loss, such as difficulty in remembering recently
learned facts. When a doctor or physician has been notified, and AD
is suspected, the diagnosis is usually confirmed with behavioral
assessments and cognitive tests, often followed by a brain scan if
available. As the disease advances, symptoms include confusion,
irritability and aggression, mood swings, language breakdown,
long-term memory loss, and the general withdrawal of the sufferer
as their senses decline (Waldemar G, Dubois B, Emre M, et al.
(January 2007). Eur J Neurol 14 (1): e1-26). Gradually, bodily
functions are lost, ultimately leading to death. Individual
prognosis is difficult to assess, as the duration of the disease
varies. AD develops for an indeterminate period of time before
becoming fully apparent, and it can progress undiagnosed for
years.
[0481] Research indicates that the disease is associated with
plaques and tangles in the brain (Tiraboschi P, Hansen L A, Thal L
J, Corey-Bloom J (June 2004). Neurology 62 (11): 1984-9). The
disease course is divided into four stages, with a progressive
pattern of cognitive and functional impairment: pre-dementia, early
dementia, moderate dementia, and advanced dementia Alzheimer's
disease is characterized by loss of neurons and synapses in the
cerebral cortex and certain subcortical regions. This loss results
in gross atrophy of the affected regions, including degeneration in
the temporal lobe and parietal lobe, and parts of the frontal
cortex and cingulate gyrus. Both amyloid plaques and
neurofibrillary tangles are clearly visible by microscopy in brains
of those afflicted by AD.
[0482] Alzheimer's disease has been identified as a protein
misfolding disease (proteopathy), caused by accumulation of
abnormally folded A-beta and tau proteins in the brain (Hashimoto
M, Rockenstein E, Crews L, Masliah E (2003) Neuromolecular Med. 4
(1-2): 21-36). Plaques are made up of small peptides, 39-43 amino
acids in length, called beta-amyloid (also written as A-beta or
A.beta.). Beta-amyloid is a fragment from a larger protein called
amyloid precursor protein (APP), a transmembrane protein that
penetrates through the neuron's membrane. APP is critical to neuron
growth, survival and post-injury repair (Priller C, et al. 2006 J.
Neurosci. 26 (27): 7212-21). In Alzheimer's disease, an unknown
process causes APP to be divided into smaller fragments by enzymes
through proteolysis (Hooper N M (April 2005) Biochem. Soc. Trans.
33 (Pt 2): 335-8). One of these fragments gives rise to fibrils of
beta-amyloid, which form clumps that deposit outside neurons in
dense formations known as senile plaques (Ohnishi S, Takano K
(March 2004) Cell. Mol. Life Sci. 61 (5): 511-24). AD is also
considered a tauopathy due to abnormal aggregation of the tau
protein. Every neuron has a cytoskeleton, an internal support
structure partly made up of structures called microtubules. Tau
protein stabilizes the microtubules when phosphorylated, and is
therefore called a microtubule-associated protein. In AD, tau
undergoes chemical changes, becoming hyperphosphorylated; it then
begins to pair with other threads, creating neurofibrillary tangles
and disintegrating the neuron's transport system (Hernandez F,
Avila J September 2007 Cell. Mol. Life Sci. 64 (17): 2219-33).
[0483] It is not known exactly how disturbances of production and
aggregation of the beta amyloid peptide gives rise to the pathology
of AD. The amyloid hypothesis traditionally points to the
accumulation of beta amyloid peptides as the central event
triggering neuron degeneration. Accumulation of aggregated amyloid
fibrils, which are believed to be the toxic form of the protein
responsible for disrupting the cell's calcium ion homeostasis,
induces apoptosis (Yankner B A, Duffy L K, Kirschner D A (October
1990) Science (journal) 250 (4978): 279-82). It is also known that
A.beta. selectively builds up in the mitochondria in the cells of
Alzheimer's-affected brains, and it also inhibits certain enzyme
functions and the utilization of glucose by neurons (Chen X, Yan S
D (December 2006) IUBMB Life 58 (12): 686-94). Various inflammatory
processes and cytokines may also have a role in the pathology of
Alzheimer's disease. Inflammation is a general marker of tissue
damage in any disease, and may be either secondary to tissue damage
in AD or a marker of an immunological response (Greig N H, Mattson
M P, Perry T, et al. (December 2004) Ann. N.Y. Acad. Sci. 1035:
290-315).
[0484] Four medications are currently approved by regulatory
agencies such as the U.S. Food and Drug Administration (FDA) and
the European Medicines Agency (EMEA) to treat the cognitive
manifestations of AD: three are acetylcholinesterase inhibitors and
the other is memantine, an N-methyl-D-aspartic acid (NMDA) receptor
antagonist. Reduction in the activity of the cholinergic neurons is
a well-known feature of Alzheimer's disease (Geula C, Mesulam M M
(1995). Alzheimer Dis Assoc Disord 9 Suppl 2: 23-28).
Acetylcholinesterase inhibitors are employed to reduce the rate at
which acetylcholine (ACh) is broken down, thereby increasing the
concentration of ACh in the brain and combating the loss of ACh
caused by the death of cholinergic neurons (Stahl S M (2000). J
Clin Psychiatry 61 (11): 813-814). Examples of the cholinesterase
inhibitors approved for the management of AD symptoms include
donepezil, galantamine, and rivastigmine. There is evidence for the
efficacy of these medications in mild to moderate Alzheimer's
disease, and some evidence for their use in the advanced stage
(Birks J, Harvey R J (2006). Cochrane Database Syst Rev (1):
CD001190). Only donepezil is approved for treatment of advanced AD
dementia. The common side effects associated with cholinesterase
inhibitors include nausea and vomiting, muscle cramps, decreased
heart rate (bradycardia), decreased appetite and weight, and
increased gastric acid production.
[0485] Donepezil, a cholinesterase inhibitor, is the most widely
used drug for Alzheimer's disease. Donepezil hydrochloride is
marketed by Eisai Inc. and Pfizer Inc. under the brand name
Aricept.RTM.. It helps increase the levels of acetylcholine. In
Alzheimer's disease there is a deficiency in acetlycholine in some
areas of the brain, which accounts for some of the symptoms of the
disease. Donepezil works by slowing down the breakdown of
acetylcholine. Donepezil is the only treatment approved by the FDA
for all stages of Alzheimer's disease: mild, moderate, and severe.
Donepezil does not cure Alzheimer's, but studies have shown that in
some patients it can improve mental function, which includes
effects on memory and daily living.
[0486] Rivastigmine tartrate by Novartis Pharmaceutical Corporation
is branded as Exelon.RTM. and Exelon.RTM. Patch. Galantamine
hydrobromide is branded as Razadyne.RTM. and Razadyne.RTM. ER by
Ortho-McNeil-Janssen Pharmaceuticals, Inc. These drugs also work by
inhibiting the breakdown of acetylcholine. They are most effective
when given in the earlier stages of Alzheimer's disease. They also
have side effects similar to donepezil.
[0487] Tacrine, a reversible cholinesterase inhibitor, also works
by slowing the breakdown of acetylcholine. Tacrine hydrochloride is
sold by Sciele Pharma, Inc. as Cognex.RTM.. Side effects of the
drug include nausea, vomiting, diarrhea, abdominal pain, skin rash,
and indigestion.
[0488] Memantine is a noncompetitive NMDA receptor antagonist first
used as an anti-influenza agent. Memantine hydrochloride is
marketed under the brands Axura.RTM. and Akatinol.RTM. by Merz
Pharmaceuticals, Namenda.RTM. by Forest Pharmaceuticals, Inc.,
Ebixa.RTM. and Abixa.RTM. by H. Lundbeck A/S and Memox.RTM. by
Unipharm Ltd. It acts on the glutamatergic system by blocking NMDA
receptors and inhibiting their overstimulation by glutamate (Lipton
S A (2006) Nat Rev Drug Discov 5 (2): 160-170). Memantine has been
shown to be moderately efficacious in the treatment of moderate to
severe Alzheimer's disease. Memantine may have increased benefit
when used in combination with donepezil, rivastigmine, galantamine,
or THA. Side effects of memantine include tiredness, dizziness,
confusion, and headache.
[0489] Antipsychotic drugs are modestly useful in reducing
aggression and psychosis in Alzheimer's patients with behavioral
problems, but are associated with serious adverse effects, such as
cerebrovascular events, movement difficulties or cognitive decline,
that do not permit their routine use (Ballard C, et al. 2009 Lancet
Neurology 8: 151).
[0490] In addition to these medicines, the American Academy of
Neurology has stated that vitamin E supplements (alpha-tocopherol)
likely delay the time to clinical worsening in patients with
Alzheimer's disease.
[0491] Glutamate is a useful excitatory neurotransmitter of the
nervous system, although excessive amounts in the brain can lead to
cell death through a process called excitotoxicity which consists
of the overstimulation of glutamate receptors. Excitotoxicity
occurs not only in Alzheimer's disease, but also in other
neurological diseases such as Parkinson's disease and multiple
sclerosis. Progesterone and vitamin D may afford neuroprotection
against excitotoxicity.
[0492] Psychosocial interventions are used as an adjunct to
pharmaceutical treatment for AD and can be classified within
behavior-, emotion-, cognition- or stimulation-oriented approaches.
Behavioral interventions attempt to identify and reduce the
antecedents and consequences of problem behaviors. This approach
has not shown success in improving overall functioning, but can
help to reduce some specific problem behaviors, such as
incontinence. Emotion-oriented interventions for treating AD
include but are not limited to reminiscence therapy, validation
therapy, supportive psychotherapy, sensory integration, also called
snoezelen, and simulated presence therapy. Reminiscence therapy
(RT) involves the discussion of past experiences individually or in
group, many times with the aid of photographs, household items,
music and sound recordings, or other familiar items from the past.
Although there are few quality studies on the effectiveness of RT,
it may be beneficial for cognition and mood. Simulated presence
therapy (SPT) is based on attachment theories and involves playing
a recording with voices of the closest relatives of the person with
Alzheimer's disease. There is preliminary evidence indicating that
SPT may reduce anxiety and challenging behaviors. Finally,
validation therapy is based on acceptance of the reality and
personal truth of another's experience, while sensory integration
is based on exercises aimed to stimulate senses. The aim of
cognition-oriented treatments, which include reality orientation
and cognitive retraining, is the reduction of cognitive deficits.
Reality orientation consists in the presentation of information
about time, place or person in order to ease the understanding of
the person about its surroundings and his or her place in them. On
the other hand cognitive retraining tries to improve impaired
capacities by exercitation of mental abilities.
Stimulation-oriented treatments include art, music and pet
therapies, exercise, and any other kind of recreational activities.
Stimulation has modest support for improving behavior, mood, and,
to a lesser extent, function.
[0493] In some embodiments, the methods of the present invention
comprise administration of chlorite or a chlorite-containing
compound in combination with other therapeutic agents and/or
interventions that are used for the treatment of Alzheimer's
disease and related complications. In one embodiment, chlorite or a
chlorite-containing compound of the present invention is used in
combination with an acetylcholinesterase inhibitor such as
donepezil, rivastigmine, galantamine, or THA. In another
embodiment, chlorite or a chlorite-containing compound of the
present invention is used in combination with an NMDA receptor
antagonist such as memantine. In another embodiment, chlorite or a
chlorite-containing compound of the present invention is used in
combination with vitamin E, progesterone or vitamin D. In other
embodiments, chlorite or a chlorite-containing compound of the
present invention is used in combination with one or more
psychosocial interventions including behavior, emotion, cognition,
and stimulation-oriented therapies.
E. Methods of Diagnois
[0494] According to an embodiment of the invention, all of the
formulations and pharmaceutical formulations described herein may
be used in methods of diagnosis of disease or disorders. Methods of
diagnosis may be combined with methods of treatment. In some
embodiments, methods of diagnosis and treatment may be integrated.
For example, in some embodiments, methods of diagnosis may be used
to evaluate the efficacy of treatment and/or whether additional
iterations of treatment are necessary. In some embodiments, methods
of treatment may be interspersed with methods of diagnosis bassed
on biomarkers to evaluate the stage of the disease.
[0495] According to an embodiment of the invention, a
macrophage-related disease is diagnosed by measuring elevated
levels of one or more biomarkers such as OPN, sCD14, and/or sCD163
in a subject. For example, the plasma levels of sCD14 and/or sCD163
are measured and then correlated against the normal plasma levels
to make or confirm a diagnosis of disease. Similarly,
responsiveness to treatment with an oxidative agent or with an
immunomodulator in combination with an oxidative agent is evaluated
by measuring reduced levels of one or more biomarkers, such as OPN,
sCD14, and/or sCD163 following treatment with an oxidative agent or
with an immunomodulator in combination with an oxidative agent.
According to certain embodiments, elevated levels of OPN, sCD14,
and/or sCD163 indicate pathology, and reduction of the levels of
the biomarkers are an indication of efficacy of the treatment with
an oxidative agent or with an immunomodulator in combination with
an oxidative agent. In certain embodiments, in vitro treatment with
an oxidative agent is performed on a sample from a subject to
determine if the subject's disease will be responsive to treatment
with an oxidative agent. In certain embodiments, one or more
bioassays are performed after treatment with the oxidative agent to
measure levels of biomarker to determine in vivo efficacy. In
various embodiments, the biomarkers of interest are sCD14 and/or
sCD163.
[0496] Methods of diagnosis according to the invention may be used
individually or in combination with other methods of diagnosis. For
example, there is no single test used to make an ALS diagnosis. An
ALS diagnosis is typically based on a person's symptoms, such as
spasticity and muscle weakness. Tests used to rule out other
diseases before making an accurate ALS diagnosis include but are
not limited to electromyography (EMG), nerve conduction velocity
(NCV), magnetic resonance imaging (MRI), blood and urine tests, and
muscle biopsy. Electromyography (EMG) is a test that uses a special
recording technique that detects electrical activity in muscles. As
muscles contract, they emit a weak electrical signal that can be
detected, amplified, and tracked, providing information about how
well the muscles are working. These responses are abnormal in cases
of ALS. Another common test measures nerve conduction velocity
(NCV). Specific abnormalities in the NCV results may suggest that
the patient has a form of peripheral neuropathy (damage to
peripheral nerves) or myopathy (muscle disease) rather than ALS.
MRI is a noninvasive procedure that uses a magnetic field and radio
waves to take detailed images of the brain and spinal cord. MRI
scans can reveal evidence of other problems that may be causing the
symptoms, such as a spinal cord tumor, a herniated disk in the
neck, syringomyelia (a cyst in the spinal cord), or cervical
spondylosis (arthritis of the neck). Blood tests may be used to
detect the presence of heavy metals such as lead in the blood.
Laboratory tests may detect abnormal proteins or hormone levels
associated with other neurological diseases. A lumbar puncture or
spinal tap may be performed to analyze the cerebrospinal fluid for
genetic abnormalities (e.g., viral, autoimmune, neurotoxic). In
order to make a definitive ALS diagnosis, a physician will
investigate the patient's full medical history and conduct a
neurological exam. The examination can be performed at regular
intervals to assess whether possible symptoms of ALS are getting
progressively worse.
[0497] In some embodiments, ALS is diagnosed by measuring chemokine
production of macrophages from a subject being screened for ALS.
For example, ALS may be diagnosed by measuring levels of sCD14
and/or sCD163 in a subject and comparing the measured levels to the
levels in non-ALS subjects. In various embodiments, efficacy of ALS
treatment may be diagnosed or monitored by measuring levels of
sCD14 and/or sCD163 in a subject and comparing such levels to
normal or to the subject's previous levels. In some embodiments,
the ALS diagnosis may be based on median CD16 expression levels in
CD14+ cells before treatment (i.e., diagnosis) or in conjunction
with treatment (i.e., monitoring efficacy of treatment). In various
embodiments, treatment may be with an oxidative agent such as
sodium chlorite, chloramine-T (including the hydrate), and/or
1,3-dichloro-5,5-dimethylhydantoin.
[0498] In some embodiments, the subjects that are treated with the
methods of the present invention are subjects who experience one or
more of the symptoms including but not limited to muscle weakness,
atrophy of muscles, hyperreflexia, and spasticity. In some
embodiments, the subjects that can be treated with the methods of
the present invention are patients who have been diagnosed with ALS
based on EMG, NCV, MRI, blood or urine tests, or muscle biopsy. In
some embodiments, methods of diagnosis and/or treatment are
directed to biomarkers for ALS, including, for example, ubiquitin,
TAR DNA-binding protein (TARDBP, TDP-43), Nogo-A, or SOD1. See, for
example, Pradat et al., Mol. Diagn. Ther., 2009:13(2):115-25,
hereby incorporated by reference.
[0499] In terms of diagnosis for Parkinson's disease (PD), there is
no specific test or marker for PD. Typically, the diagnosis is
based on medical history and neurological examination conducted by
interviewing and observing the patient in person, which may include
using the Unified Parkinson's Disease Rating Scale. A radiotracer
for SPECT scanning machines called DaTSCAN is specialized for
diagnosing dopamine loss characteristic of Parkinson's disease. The
disease can be difficult to diagnose accurately, especially in its
early stages due to symptom overlap with other causes of
Parkinsonism. Physicians may need to observe the person for some
time until it is apparent that the symptoms are consistently
present. CT and MRI brain scans of people with PD are normal and
therefore, not useful for diagnosis. However, doctors may sometimes
request brain scans or laboratory tests in order to evaluate for
other diseases that may produce signs of Parkinsonism.
[0500] To diagnose PD, the physician will perform a standard
neurological examination, involving various simple tests of
reactions, reflexes, and movements. Diagnosis of PD generally
depends on the presence of at least two of the three major signs:
tremor at rest, rigidity, and bradykinesia, as well as the absence
of a secondary cause, such as antipsychotic medications or multiple
small strokes in the regions of the brain controlling movement.
Patients tend to be most aware of tremor and bradykinesia, and less
so of rigidity. Bradykinesia is tested by determining how quickly
the person can tap the finger and thumb together, or tap the foot
up and down. Tremor is determined by simple inspection. The
physician assesses rigidity by moving the neck, upper limbs, and
lower limbs while the patient relaxes, feeling for resistance to
movement. Postural instability is tested with the "pull test," in
which the examiner stands behind the patient and asks the patient
to maintain their balance when pulled backwards. The examiner pulls
back briskly to assess the patient's ability to recover, being
careful to prevent the patient from falling. The examination also
involves recording a careful medical history, especially for
exposure to medications that can block dopamine function in the
brain.
[0501] In some embodiments, the subjects that can be treated with
the methods of the present invention are patients who experience
one or more of the symptoms including but not limited to tremor of
hands, arms, legs, jaw and face, stiffness or rigidity of the arms,
legs and trunk, slowness of movement, poor balance and
coordination, and postural instability. In some embodiments, the
subjects that can be treated with the methods of the present
invention are patients who have been diagnosed with Parkinson's
disease by a physician. In some embodiments, the subjects that can
be treated with the methods of the present invention are patients
who have not been diagnosed with Parkinson's disease but are
experiencing symptoms of PD.
[0502] There is currently no single test that accurately diagnoses
Alzheimer's disease. Diagnosing Alzheimer's involves several types
of evaluations. Evaluations commonly performed for diagnosis of
Alzheimer's disease include but are not limited to: medical history
which comprises an interview or questionnaire to identify past
medical problems, difficulties in daily activities and prescription
drug use, among other things. The doctor may wish to speak to a
close family member to supplement information. Physical examination
can include evaluation of hearing and sight, as well as blood
pressure and pulse readings. Standard laboratory tests may include
blood and urine tests designed to help eliminate other possible
conditions. These will measure things like blood count, thyroid and
liver function, and levels of glucose and other blood-based
indicators of illness. A depression screening is also typically
conducted. In some cases, a small sample of spinal fluid may be
collected for testing. Neuropsychological testing can be performed
in which doctors use a variety of tools to assess memory,
problem-solving, attention, vision-motor coordination and abstract
thinking, such as performing simple mental calculations. The goal
is to better characterize the types of cognitive symptoms present,
which might provide clues to the underlying cause. Brain-imaging
scan which is a "structural" brain scan such as CT or MRI is
recommended to rule out brain tumors or blood clots in the brain as
the reason for symptoms. Other brain-imaging techniques might be
able to identify telltale signs of early Alzheimer's reliably
enough to be used as diagnostic tools.
[0503] In some embodiments, the subjects that are treated with the
methods of the present invention are patients who experience one or
more of the symptoms of Alzheimer's disease including but not
limited to disturbances in short-term memory, problems with
attention and spatial orientation, changes in personality, language
difficulties and unexplained mood swings. In some embodiments, the
subjects that are treated with the methods of the present invention
have been diagnosed with Alzheimer's disease based on one or more
diagnostic tests, including but not limited to those tests
disclosed herein. In some embodiments, the subjects that are
treated with the methods of the present invention are patients who
have not been diagnosed with Alzheimer's disease but are
experiencing symptoms of Alzheimer's disease. In some embodiments,
the subjects that are treated with the methods of the present
invention have been diagnosed with Stage 1 (mild) Alzheimer's
disease. In some embodiments, the subjects that are treated with
the methods of the present invention have been diagnosed with Stage
2 (moderate) Alzheimer's disease. In some embodiments, the subjects
that are treated with the methods of the present invention have
been diagnosed with Stage 3 (severe) Alzheimer's disease.
F. Kits and Articles of Manufacture
[0504] Unless the context makes otherwise clear, all of the
formulations and pharmaceutical formulations described herein may
be used in the kits described herein. In some embodiments, the kits
are intended for administration of an oxidative agent, including
without limitation a chlorite or a chlorite-containing agent, or
pharmaceutical formulations comprising such oxidative agents. In
some embodiments, the kits are intended for administration of an
immunomodulatory agent, including without limitation an
immunosuppressive agent, or pharmaceutical formulations comprising
such immunomodulatory agents. The kits may include a unit dosage
amount of the agents or formulations as described herein. In some
variations, the kits comprise suitable packaging. In some
variations, the kits comprise instructions for use of the oxidative
and/or immunomodulatory agent. In a non-limiting example, the kit
may contain instructions for using chlorite formulations to treat
macrophage related disorders. Accordingly, the kits may be used for
any of the treatment methods described herein, and in some
embodiments contain suitable instructions for practicing any of the
treatment methods described herein. In some embodiments, the kits
are used to treat any one or more of the diseases or conditions
described herein. Kits may also comprise an aid to administration
of the oxidative or immunomodulatory agent formulation, such as an
inhaler, spray dispenser (e.g., nasal spray), syringe for injection
or pressure pack for capsules, tablets, or suppositories.
[0505] The chlorite formulations described herein may be assembled
in the form of kits. In some variations the kit provides the
chlorite and reagents to prepare an aqueous chlorite formulation
for administration. In some embodiments, the formulations in the
kits of the invention comprise chlorite-containing agents,
non-chlorite oxidative agents, or immunosuppressive agents. In some
variations the formulation is an aqueous solution. In some
variations the formulation is a sterile solution. In some
variations, a kit provides a pharmaceutically acceptable diluent,
either already mixed with the formulations or formulations
described herein or provided in a separate container from the
formulations or pharmaceutical formulations described herein. In
some variations, the diluent is a saline solution. In some
variations, the composition comprises a dry (such as lyophilized)
composition that can be reconstituted or dissolved to form the
formulations or pharmaceutical formulations described herein. When
the formulation is in a dry form, the kit may comprise one or more
of a pharmaceutically acceptable solvent, diluent, and a pH
adjusting agent, either separately from or as part of the diluent.
In some variations, a kit or article of manufacture comprises a dry
form of the active agent, e.g., chlorite, a pharmaceutically
acceptable solvent, and pH adjusting agent. In some variations the
pH adjusting agent is incorporated into the solvent. In some
variations, a kit or article of manufacture comprises the active
agent in dry form and a pharmaceutically acceptable diluent. In
some variations the pH adjusting agent is incorporated into the
diluent. In some variations, the formulations or pharmaceutical
formulations described herein are sterile, reconstituted
formulations. In some variations, the formulations or
pharmaceutical formulations described herein are sterile,
reconstituted formulations in unit dosage form. In some variations,
the formulations or pharmaceutical formulations described herein
are sterile, reconstituted formulations in unit dosage form in
suitable packaging.
[0506] The kit may contain a device for administration or for
dispensing the compositions, including, but not limited to one or
more syringes, pipettes, transdermal patches, or inhalants.
[0507] The kit may include other therapeutic compounds or
formulations for use in conjunction with the formulations described
herein. These compounds may be provided in a separate form, or
mixed with the oxidative and/or immunomodulatory agent formulations
or pharmaceutical formulations described herein. As a non-limiting
example, a kit may comprise a chlorite formulation and a
formulation for metformin. When the therapeutic agent is contained
in a different formulation than the oxidative and/or
immunomodulatory agent of the present invention, they can be
administered sequentially or substantially simultaneously.
[0508] In some variations the kit includes instructions for
preparation and administration of the formulation. In some
variations the kit includes instructions as to side effects of the
formulation. In another variation the kit optionally includes any
other relevant information. The instructions may be in any suitable
format, including, but not limited to, printed matter, videotape,
computer readable disk, or optical disc. The instructions may be
located inside the housing or outside the housing, and may be
printed on the interior or exterior of any surface forming the
housing that renders the instructions legible.
[0509] Described herein are kits for treating an individual who
suffers from or is susceptible to a macrophage related disease
treatable by the oxidative agent, for example, chlorite
formulations described herein, comprising a container comprising a
unit dosage amount of a chlorite formulation as described herein,
and instructions for use. Further described herein are kits for
treating an individual who suffers from or is susceptible to a
macrophage related disease treatable by immunomodulatory agents,
for example, immunosuppressants, comprising a container comprising
a unit dosage amount of the agent and instructions for use. The
container may be any of those known in the art and appropriate for
storage and delivery of oral, intravenous, systemic, parenteral,
rectal, urethral, transdermal, or inhalation formulations.
[0510] Kits may also be provided that contain sufficient dosages of
the oxidative agent formulation to provide effective treatment for
an individual for an extended period, including but not limited to
any of about a week, about 2 weeks, about 3 weeks, about 4 weeks,
about 6 weeks or about 8 weeks or more.
[0511] Also provided herein are articles of manufacture comprising
the formulations or pharmaceutical formulations described herein,
or unit dosage forms in suitable packaging, including but not
limited to vials or vessels, including but not limited to sealed
vials or vessels and sterile sealed vials or vessels. Non-limiting
examples of suitable packaging for the formulations and
pharmaceutical formulations described herein are known in the art,
and include, for example, any of vials (such as sealed vials),
vessels (such as sealed vessels), ampules, bottles, jars, flexible
packaging (e.g., sealed Mylar or plastic bags), and the like. Such
packaging may optionally limit the amount of light to which the
formulation is exposed. These articles of manufacture may further
be sterilized and/or sealed.
[0512] The invention is further illustrated by the following
non-limiting examples.
EXAMPLES
Example 1
Purification of Chlorite
[0513] This procedure was performed in diminished light, e.g., with
overhead lights off, and out of direct sunlight.
[0514] Sodium Chlorite (80 wt %, Sigma-Aldrich lot #09911CD) was
dissolved in 1000 mL of distilled water. The flask was mounted to a
rotary evaporator, and the bath temperature set to 70.degree. C.
Vacuum was applied, and increased until the water began to distill
over in a controlled manner. The vacuum was applied until the
mixture put down a load of solids, and 550 mL of water had
distilled over. Using a coarse sintered glass funnel, the solids
were removed by suction filtration of the hot solution. These
solids were mostly sodium chloride.
[0515] The filtrate was stored at -25.degree. C. for a period of
time sufficient to precipitate the chlorite (approximately 24
hours). The entire mixture froze solid. The frozen mixture was
broken up and centrifugally filtered while cold. Purified sodium
chlorite was collected as the frozen solid melted. The centrifuge
had a 12-inch stainless steel basket, 50 micron polypropylene bag,
and was run at 2000 rpm. HPLC analysis using an ion-separating
column and ion detector showed 99.04% purity. The material is
presumed to be a mixture of hydrate and non-hydrate.
Example 2
Purification of Chlorite
[0516] The method described in Example 1 was performed, but using
coarse sintered glass suction filtration rather than centrifugal
filtration for the cold filtration. After the first filtration,
chlorite purity after the first crystallization was 91.9%. The
crystallization step was repeated a second time. After the second
recrystallization/suction filtration, the chlorite was 99.5%
pure.
[0517] In one method, a suspension of NaClO.sub.2 (technical grade,
80% purity) is triturated with water at 17-25.degree. C., the
suspension is stored at -25.degree. C. to -5.degree. C., and the
cold mixture is filtered. The series of steps is repeated until the
sodium chlorite assay shows .gtoreq.99.0% area under the curve the
ion chromatography.
[0518] FIG. 5 shows a Thermo-Gravimetric Analysis (TGA) of a sample
of sodium chlorite purified according to the invention. The
thermogram shows the loss of a total of 40.0% weight from ambient
temperature to about 160.degree. C. FIG. 6 shows an X-ray powder
diffraction (XRPD) pattern for a sample of sodium chlorite purified
according to the invention. The XRPD pattern indicates that the
material is crystalline.
Example 3
Adjustment of Chlorite Formulation pH
[0519] To prepare a chlorite formulation at a lower pH, sodium
chlorite purified by the method of Example 2 was dissolved in
distilled water and stirred using a magnetic stirrer. A calibrated
pH probe was put in the solution. Small amounts of monosodium
phosphate monohydrate were added, until the pH reached and
stabilized at 7.62. In the event of the pH drifting lower than the
target pH, the pH can be adjusted back with 0.1 N NaOH.
[0520] This solution was sampled, and assayed for sodium chlorite
content by HPLC. Column: Novosep A-2 Alltech 250.times.4 mm;
eluant: 3.6 mM sodium carbonate. Rate: 0.8 mL/min. Detected with a
suppressed Alltech 650 conductivity detector. Quantitation was
performed by standard iodimetry. See Inorganic Syntheses, section
under Chlorine (IV) Oxide; Sodium Chlorite analysis, p. 156. The
concentration was determined to be 1.36 M. To prepare a 4.25 wt %
solution (0.47 M), 200 mL were diluted to 580 mL.
Example 4
Treatment of a Macrophage Related Disease-Multiple Sclerosis
[0521] Experimental Autoimmune Encephalomyelitis (EAE) is a disease
that is given to laboratory animals that produces symptoms similar
to those of multiple sclerosis (MS) in humans. It is mostly used in
mice and rats, but can also be produced in monkeys, rabbits and
guinea pigs.
[0522] Female C57BL/6, mice 5-8 weeks old are immunized
subcutaneously with 200 .mu.g of MOG.sub.35-55 peptide emulsified
in CFA supplemented with 5 mg/ml of Mycobacterium tuberculosis. The
mice receive intraperitoneal injections with 250 ng pertussis toxin
(Sigma-Aldrich, St. Louis, Mo., USA) at the time of immunization
and 48 hours later. After 7 days, the mice receive an identical
booster immunization with MOG/CFA without pertussis toxin. Clinical
disease of EAE usually commences between day 16 and day 20 after
immunization.
[0523] Macrophages are important effector cells involved in the
pathogenesis of demyelination in multiple sclerosis. These EAE mice
will be administered chlorite to reduce or inhibit macrophage
activation and thereby ameliorating the disease. The mice will be
treated in two ways: 1) short term--i.e., just long enough to
observe the desired change in macrophage phenotype--which may
require only one or two injections of chlorite; and 2) longer term
monitor disease progression. Setting up the model, treating the
mice, and monitoring CNS lesions and neurological degeneration are
straightforward to those of skill in the art.
[0524] The mice are scored four times per week as follows: 0, no
detectable signs of EAE; 0.5, limp distal tail; 1, complete limp
tail; 1.5, limp tail and hind limb weakness; 2, unilateral partial
hind limb paralysis; 2.5, bilateral partial hind limb paralysis; 3,
complete bilateral hind limb paralysis; 3.5, complete hind limb
paralysis and unilateral forelimb paralysis; 4, total paralysis of
both forelimbs and hind limbs; 5, death. Mice scoring greater than
4 but less than 5 are euthanized.
[0525] The results will suggest that chlorite is effective in
treating EAE in mice.
Example 5
Evaluation of Oxidative Drugs for Immune Regulating Activity in
Vitro and in Vivo
[0526] A panel of in vitro tests shows activities of drug in dose
dependent manner duplicating in vitro activities of reference drug
that has in vivo activities. The goal of this example is to
duplicate the in vivo regulatory activities of WF10 chlorite.
Examplary in vivo regulatory activities of WF10 chlorite include
but are not limited to regulation of monocyte HLA-DR expression,
regulation of TNF-.alpha. RNA expression, and restoration of
macrophage phagocytic function in vivo. Oxidative drugs are tested
as compared to WF10. One way of evaluating oxidative drugs for
equivalency is to test them in dose ranging studies for effects on
monocyte cell surface antigen expression and effects on secretory
molecules.
[0527] In vitro drug study: Treatment of PBMC (peripheral blood
mononuclear cells) with chlorite or WF 10.
[0528] The purpose of this study is to assess the effect of
chlorite and WF10 (a chlorite-based compound) on primary cultures
of peripheral blood mononuclear cells (PBMC) and compare the
differences between chlorite and WF10. The procedure is performed
in a biological safety cabinet using standard aseptic techniques
and universal precautions for handling human blood samples well
known to one skilled in the art.
[0529] PBMCs are cultured in RPMI-1640 medium with 2.0 g/L Glucose,
0.3 g/L L-glutamine, and 2.0 g/L NaHCO.sub.3 containing 10% fetal
bovine serum and 11 g/L sodium pyruvate. Peripheral blood
mononuclear cells (PBMC) are isolated from Heparinized Blood and
collected in BD Vacutainer Blood Collection Tubes with sodium
heparin according to standard techniques known in the art and the
manufacturer's protocols. PBMC concentration is adjusted to
1.times.10.sup.6 cells/mL in the culture media for chlorite/WF 10
treatment.
[0530] In terms of the treatment of PBMC with chlorite or WF10,
PBMCs are isolated and cell density is adjusted to 1.times.10.sup.6
cells/mL in the culture media. PBMC suspensions (1.times.10.sup.6
cells/mL) are transferred to polypropylene tubes: .ltoreq.2 mL cell
suspensions are transferred into 12.times.75 mm polypropylene
tubes; >3 mL cell suspensions are transferred to 50mL
prolypropylene tubes. For dose response experiments, 1 tube of
PBMCs is treated for each dose of chlorite or WF10, and each dose
is assayed with triplicate culture tubes. Chlorite and WF10 are
prepared as shown in the tables below: chlorite/WF10 diluted
solutions are prepared immediately before use.
TABLE-US-00002 Initial Intermediate Final concentration
concentration concentration (.mu.M) (.mu.M) (.mu.M) Chlorite Stock
concentration = 57 mM, stored in the dark at 4.degree. C. 57000
1:9.5 (210.5 .mu.L of 6000 1:10 (100 .mu.L of 600 chlorite stock +
6000 .mu.M chlorite 1789.5 .mu.L media) solution + 900 .mu.L of
cell suspension) 6000 1:2 (1000 .mu.L of 3000 1:10 (100 .mu.L of
300 6000 .mu.M chlorite 3000 .mu.M chlorite solution + 1000 .mu.L
solution + 900 .mu.L of media) cell suspension) 3000 1:3 (700 .mu.L
of 3000 .mu.M 1000 1:10 (100 .mu.L of 100 chlorite solution + 1000
.mu.M chlorite 1400 .mu.L media) solution + 900 .mu.L of cell
suspension) 3000 1:10 (100 .mu.L of 300 1:10 (100 .mu.L of 30 3000
.mu.M chlorite 300 .mu.M chlorite solution + 900 .mu.L solution +
900 .mu.L of media) cell suspension) 1000 1:10 (100 .mu.L of 100
1:10 (100 .mu.L of 10 1000 .mu.M chlorite 100 .mu.M chlorite
solution + 900 .mu.L solution + 900 .mu.L of media) cell
suspension) WF10 Stock concentration = 62 mM, stored in the dark at
4.degree. C. 62000 1:10.333 (241.9uL of 6000 1:10 (100 .mu.L of 600
WF10 stock + 6000 .mu.M WF10 2258.1 .mu.L media) solution + 900
.mu.L of cell suspension) 6000 1:2 (1000 .mu.L of 3000 1:10 (100
.mu.L of 300 6000 .mu.M WF10 3000 .mu.M WF10 solution + 1000 .mu.L
solution + 900 .mu.L of media) cell suspension) 3000 1:3 (700 .mu.L
of 3000 .mu.M 1000 1:10 (100 .mu.L of 100 WF10 solution + 1000
.mu.M WF10 1400 .mu.L media) solution + 900 .mu.L of cell
suspension) 3000 1:10 (100 .mu.L of 300 1:10 (100 .mu.L of 30 3000
.mu.M WF10 300 .mu.M WF10 solution + 900 .mu.L solution + 900 .mu.L
of media) cell suspension) 1000 1:10 (100 .mu.L of 100 1:10 (100
.mu.L of 10 1000 .mu.M WF10 100 .mu.M WF10 solution + 900 .mu.L
solution + 900 .mu.L of media) cell suspension)
[0531] Varying doses of chlorite or WF10 are added to each tube of
PBMC suspensions and mixed gently by slowly pipeting up and down a
few times. For the control tube containing no drug, the same volume
of blank media is added instead. Cell suspensions are incubated in
a humidified incubator at 37.degree. C. with a 5% CO.sub.2
atmosphere for 3 days. On day 3, PBMCs are centrifuged at 1520 RPM
(300.times.g) for 10 minutes at 25.degree. C. and cell culture
supernatants are collected and stored at -80.degree. C. for
cytokine assays. Cell pellets are washed once by
PBS(Mg.sup.+2/Ca.sup.+2 Free) for immunophenotype flow cytometric
analysis.
[0532] Immunophenotype flow cytometry assay: Flow cytometry is a
standard technique for assaying immunophenotype and it is well
known to one of ordinary skill in the art. Briefly, PBMCs treated
as described hereinabove are resuspended in PBS
(Mg.sup.+2/Ca.sup.+2Free) to make the final concentration
.about.0.5-1.times.10.sup.6 cells/mL, and aliquoted at 100 .mu.l
cell suspendsion into 12.times.75 mm polystyrene tube. PBMCs are
stained with CD16-PE and CD14-PerCP for 20 minutes in the dark at
room temperature. Negative controls consist of aliquots stained
with isotype IgG-PE and IgG-PerCP (all staining are performed per
manufacturer's specifications). The stained PBMCs are washed by
adding 2mL of PBS(Mg.sup.+2/Ca.sup.+2 Free), and then centrifuged
at 1520 RPM (300.times.g) for 10 minutes at 25.degree. C. Cells are
fixed by add 0.5 mL of fixative solution (1%
paraformaldehyde/NaN.sub.3/PBS). Fluorescent emission is measured
using FACSCAN flow cytometor (Becton-Dickinson).
[0533] For the culture supernatant quantitative cytokine assay,
some of the PBMC culture supernatants are sent out to a commercial
laboratory, AssayGate, Inc, Ijamsville Md., USA, for cytokine
quantitative analysis. Other quantitative cytokine assays are
performed with commercially available ELISA kits according to the
manufacturer's instructions.
[0534] FIGS. 1-4 illustrate examples testing two different
formulations of chlorite or WF 10. Chlorite and WF10 regulate cell
surface and secreted antigens of monocytes in vitro. Cell surface
antigen CD16 was measured on CD14+ cells, i.e., monocytes, for both
normal peripheral blood mononuclear cells (PBMC) and ALS. For
measuring secreted molecules, normal PBMC culture supernatants were
collected and measured three days after exposure to the oxidative
drug in various concentrations. Levels of secreted molecules such
as osteopontin, MMP-9, TNF-.alpha., IL-6, IL-8, and MCP-1 were
measured. All results were normalized to chlorite
concentration.
[0535] FIG. 1 shows median CD16 expression levels on CD14+ cells
after exposure of five samples of normal PBMCs to WF 10 (triangle)
or chlorite (square) at various concentrations for three days. As
the concentration of WF10 or chlorite increased, the levels of
CD14CD16 on PBMCs in all five samples were decreased. FIG. 2 shows
that treatment of five ALS blood samples with a chlorite
formulation containing 150 .mu.M chlorite caused down-regulation of
CD16 surface expression on CD14+ cells, i.e., monocytes. Cell
surface expression of a molecule can be measured by a variety of
techniques known in the art, for example, flow cytometry.
[0536] Chlorite also regulates the secretion of molecules by
monocytes. Monocytes were exposed to chlorite or WF 10 at various
concentrations for three days. Culture supernatants were collected
and the levels of secreted molecules in the supernatants were
measured by enzyme-linked immunosorbent assay (ELISA), which is
well known to one of ordinary skill in the art. An example of such
secreted molecule is osteopontin, which is a macrophage chemotactic
protein. As FIG. 3 shows, osteopontin secretion by monocytes was
down-regulated by both WF10 (OPN-W) and chlorite (OPN-C) to a
similar extent as increasing concentrations of chlorite inhibited
osteopontin secretion, suggesting an effect of chlorite on
macrophage activation and accumulation.
[0537] Cytokine secretion by PMBCs upon exposure to chlorite was
also measured. PBMCs from ALS blood samples were exposed to
chlorite or WF10 for three days. Supernatants were harvested and
levels of cytokines including monocyte chemotactic protein-1
(MCP-1), OPN, matrix metallopeptidase 9 (MMP-9), and IL-6 were
determined by ELISA. As shown in FIG. 4, chlorite downregulated the
secretion of MCP-1, OPN, and MMP-9, while it did not have
significant effect on IL-6 secretion as compared to the control.
Chlorite and WF10 had similar effects on the cytokine secretion by
PBMCs from ALS blood samples. As shown in FIG. 12, ALS PBMCs were
treated with 300 micromolar sodium chlorite for 3 days, and
supernatant mean levels of sCD14 and sCD163 were determined by
ELISA (n=13), which showed that chlorite downregulated the
secretion of sCD14 and sCD163.
[0538] CD14 cells typically require C16+ differentiation state to
migrate from blood into tissues. FIG. 8 shows the effect of sodium
chlorite (n=4) and various additional compounds at 300 micromolar
concentration on CD16 expression in CD14+ cells as well as monocyte
toxicity. Experimental conditions were generally as described
above. Sodium chlorite and N-chloro compounds decreased CD16
expression. Toxicity data showed the reduction in % CD14 monocytes
according to the following qualitative scale: "No" was an increase
or no change in % CD14 monocytes following treatment with the
compound; +/- was less than 5% reduction; + was reduction between
11-20%; ++ was reduction between 21-30%; +++ was reduction between
31-40%; and ++++ was reduction between 41-50%.
[0539] As shown, chlorite affected both cell surface antigen
expression and cytokine secretion by monocytes.
Example 6
Treatment of Type II Diabetes
[0540] Obese mice develop insulin resistance and glucose
intolerance equivalent to Type II diabetes. After being placed on a
high fat/high calorie diet, the mice experience an inflammatory
immune response in the visceral adipose tissue (VAT). Without being
bound by theory, the inflammatory response may be a causative agent
of the disease because the inflammatory response and the disease
can be reversed for at least several months by brief systemic
treatment with an anti-CD3 (anti-T cell) antibody. The immune cell
content in human visceral fat of obese patients appears very
similar to that seen in obese mice. Inflammatory macrophages appear
in the VAT of obese diabetic mice and these cells appear to revert
to non-inflammatory macrophages after anti-CD3 antibody treatment,
suggesting that the effect of anti-CD3 antibody treatment may be at
least partially mediated through the change in macrophages.
[0541] The diet induced obese (DIO) mouse model is an excellent
model of Type II diabetes. To create the model, normal mice, e.g.,
C57Black/6, are put on a high fat/high calorie diet. DIO mice will
be administered chlorite to reverse their macrophage phenotype and
the disease. The mice will be treated in two ways: 1) short
term--i.e., just long enough to observe the desired change in
macrophage phenotype--which may require only one or two injections
of chlorite; and 2) longer term monitor disease progression.
Setting up the model, treating the mice, and monitoring glucose
tolerance and insulin sensitivity are straightforward to those of
skill in the art. VAT will be assessed for cellular analysis using
flow cytometry.
Example 7
Treatment of ALS
[0542] Laboratory models of amyotrophic lateral sclerosis (ALS) can
help understand the basic process of the disease, with an eye
towards developing new therapies for ALS. The mainstay of animal
model for ALS has been a mouse that bears the mutated human gene
associated with familial ALS. Mutation of the SOD1 gene can produce
many aspects of ALS. The mouse bearing the human gene for mutant
SOD1 was the first lab model clearly linked to ALS based on a known
cause of the disease. But other models are now available or being
designed. A newer rodent model, an ALS rat, also is engineered to
express human mutant SOD1. The rat is larger and surgery is easier
for applications such as stem cell transplants and other approaches
that require injections into the spinal cord. The worm, fish and
the fly models of ALS will be valuable as tools as they may offer a
basic and simple biology. Cell based tests that reflect the disease
process in ALS can rapidly report on the potential of new molecules
to serve as therapeutics.
[0543] Animal models of ALS provide an opportunity to study this
incurable and fatal human disease both clinically and
pathologically (Pioro E P, Mitsumoto H Clin Neurosci. 1995-1996;
3(6):375-85). Four natural disease models have been most
extensively studied, including three mouse models: motor neuron
degeneration (Mnd), progressive motor neuronopathy (pmn), wobbler,
and one canine model: hereditary canine spinal muscular atrophy
(HCSMA). The wobbler mouse has been the most extensively studied of
these models with analyses of clinical, pathological (perikaryon,
axon, muscle), and biochemical features. Experimentally induced ALS
animal models have allowed controlled testing of various
neurotoxic, viral and immune-mediated mechanisms. Molecular
techniques have recently generated mouse models in which genes
relevant to the human disease or motor neuron biology have been
manipulated. The most clinically relevant of these is a transgenic
mouse overexpressing the mutated SOD1 gene of FALS patients, which
has already provided significant insights into mechanisms of motor
neuron degeneration in this disease. Because no single animal model
perfectly reflects all the clinical and pathological
characteristics of ALS, study of selected features from the most
relevant models will contribute to a better understanding of the
pathogenesis and/or etiology of this disease.
[0544] The transgenic mouse overexpressing the mutated SOD1 gene of
FALS patients is an excellent model of ALS. Such ALS mice bearing
the mutant human SOD1 gene will be administered chlorite to reverse
their macrophage phenotype and the disease. The mice will be
treated in two ways: 1) short term--i.e., just long enough to
observe the desired change in macrophage phenotype--which may
require only one or two injections of chlorite; and 2) longer term
monitor disease progression. Setting up the model, treating the
mice, and monitoring neurological signs including motor activities
and disease-associated behavioral changes are straightforward to
those of skill in the art. VAT will be assessed for cellular
analysis using techniques such as flow cytometry.
Example 8
Blood Monocyte Migration
[0545] Two chamber migration studies were performed with
chemoattractant in the lower chamber and normal PBMCs in the upper
chamber (represented as circles), separated by a membrane, as shown
in FIG. 9. After four hours, the cells captured by the membrane
were normalized to a background score where no chemoattractant was
present in the lower chamber to yield a chemotaxis index. FIG. 10
shows the results of normal macrophage culture supernatant (a
10.times. dilution of 3-day supernatant) as chemoattractant in the
left bar compared with ALS patient macrophage culture supernatant
(a 10.times. dilution of 3-day supernatant) as the chemoattractant.
The results show that ALS macrophages make chemokines that attract
PBMCs. FIG. 11 shows the results of adding 300 micromolar sodium
chlorite (4 hours) to PBMCs versus untreated PBMCs. Untreated PBMCs
migrated towards 10% FBS (fetal bovine serum containing chemokines)
at a greater chemotaxis index than sodium chlorite treated PBMCs
(right bar of FIG. 11). The results show that sodium chlorite
treatment of normal PBMCs blocked the migration response to
chemoattractants.
[0546] While preferred embodiments of the present invention have
been shown and described herein, it will be clear to those skilled
in the art that such embodiments are provided by way of example
only. Numerous variations, changes, and substitutions will now
occur to those skilled in the art without departing from the
invention. It should be understood that various alternatives to the
embodiments of the invention described herein may be employed in
practicing the invention. It is intended that the following claims
define the scope of the invention and that methods and structures
within the scope of these claims and their equivalents be covered
thereby.
* * * * *
References