WO2014046722A1 - Treatment compositions - Google Patents
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- WO2014046722A1 WO2014046722A1 PCT/US2013/028420 US2013028420W WO2014046722A1 WO 2014046722 A1 WO2014046722 A1 WO 2014046722A1 US 2013028420 W US2013028420 W US 2013028420W WO 2014046722 A1 WO2014046722 A1 WO 2014046722A1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/14—Alkali metal chlorides; Alkaline earth metal chlorides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
- A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
- A61K31/216—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acids having aromatic rings, e.g. benactizyne, clofibrate
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/42—Oxazoles
- A61K31/421—1,3-Oxazoles, e.g. pemoline, trimethadione
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/42—Oxazoles
- A61K31/422—Oxazoles not condensed and containing further heterocyclic rings
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
- A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/557—Eicosanoids, e.g. leukotrienes or prostaglandins
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/46—Treatment of water, waste water, or sewage by electrochemical methods
- C02F1/461—Treatment of water, waste water, or sewage by electrochemical methods by electrolysis
- C02F1/46104—Devices therefor; Their operating or servicing
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/46—Treatment of water, waste water, or sewage by electrochemical methods
- C02F1/461—Treatment of water, waste water, or sewage by electrochemical methods by electrolysis
- C02F1/46104—Devices therefor; Their operating or servicing
- C02F1/4618—Devices therefor; Their operating or servicing for producing "ionised" acidic or basic water
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/46—Treatment of water, waste water, or sewage by electrochemical methods
- C02F1/461—Treatment of water, waste water, or sewage by electrochemical methods by electrolysis
- C02F1/467—Treatment of water, waste water, or sewage by electrochemical methods by electrolysis by electrochemical disinfection; by electrooxydation or by electroreduction
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/46—Treatment of water, waste water, or sewage by electrochemical methods
- C02F1/469—Treatment of water, waste water, or sewage by electrochemical methods by electrochemical separation, e.g. by electro-osmosis, electrodialysis, electrophoresis
- C02F1/4698—Treatment of water, waste water, or sewage by electrochemical methods by electrochemical separation, e.g. by electro-osmosis, electrodialysis, electrophoresis electro-osmosis
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- C—CHEMISTRY; METALLURGY
- C25—ELECTROLYTIC OR ELECTROPHORETIC PROCESSES; APPARATUS THEREFOR
- C25B—ELECTROLYTIC OR ELECTROPHORETIC PROCESSES FOR THE PRODUCTION OF COMPOUNDS OR NON-METALS; APPARATUS THEREFOR
- C25B9/00—Cells or assemblies of cells; Constructional parts of cells; Assemblies of constructional parts, e.g. electrode-diaphragm assemblies; Process-related cell features
- C25B9/17—Cells comprising dimensionally-stable non-movable electrodes; Assemblies of constructional parts thereof
- C25B9/19—Cells comprising dimensionally-stable non-movable electrodes; Assemblies of constructional parts thereof with diaphragms
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/46—Treatment of water, waste water, or sewage by electrochemical methods
- C02F1/461—Treatment of water, waste water, or sewage by electrochemical methods by electrolysis
- C02F1/46104—Devices therefor; Their operating or servicing
- C02F1/46109—Electrodes
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2201/00—Apparatus for treatment of water, waste water or sewage
- C02F2201/46—Apparatus for electrochemical processes
- C02F2201/461—Electrolysis apparatus
- C02F2201/46105—Details relating to the electrolytic devices
- C02F2201/4611—Fluid flow
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2201/00—Apparatus for treatment of water, waste water or sewage
- C02F2201/46—Apparatus for electrochemical processes
- C02F2201/461—Electrolysis apparatus
- C02F2201/46105—Details relating to the electrolytic devices
- C02F2201/46115—Electrolytic cell with membranes or diaphragms
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2201/00—Apparatus for treatment of water, waste water or sewage
- C02F2201/46—Apparatus for electrochemical processes
- C02F2201/461—Electrolysis apparatus
- C02F2201/46105—Details relating to the electrolytic devices
- C02F2201/46155—Heating or cooling
Definitions
- compositions useful for the treatment of diseases related to oxidative stress and reduced mitochondrial DNA are disclosed.
- RS/ROS are produced in large amounts in the cell during the metabolism of sugar, specifically by the oxidative phosphorylation process in the mitochondria.
- Embodiments include a composition comprising a redox signaling agent (RXN) and a Peroxisome Proliferator-Activated Receptor (PPAR) agonist.
- RXN redox signaling agent
- PPAR Peroxisome Proliferator-Activated Receptor
- the PPAR agonist comprises a Free Fatty Acid (FFA).
- FFA Free Fatty Acid
- the PPAR agonist comprises an eicosanoid, or a thazolidinedione, or a fibrate.
- the PPAR agonist comprises a dual agonist such as aleglitazar, muraglitazar, or tesaglitazar.
- Embodiments include the use of a composition comprising RXN and a PPAR agonist in the treatment of a PPAR-mediated disease such as diabetes mellitus, including type 1 diabetes, type 2 diabetes, gestational diabetes, obesity, or cancer.
- a PPAR-mediated disease such as diabetes mellitus, including type 1 diabetes, type 2 diabetes, gestational diabetes, obesity, or cancer.
- Embodiments useful in the treatment of cancer can comprise a first therapy which activates a PPAR pathway and at least one other agent wherein the at least one other agent does not activate a PPAR pathway, such as radiation or a chemotherapeutic agent.
- Embodiments useful in the treatment of cancer can comprise a first therapy which mobilizes Free Fatty Acids (FFAS) and at least one other agent wherein the at least one other agent does not mobilize FFAs, such as radiation.
- FFAS Free Fatty Acids
- Embodiments include a method of treating an oxidative stress related disorder, such method including administering a composition including at least one species selected from 0 2 , H 2 , Cl 2 , OCr, HOCI, NaOCI, HCI0 2 , CI0 2 , HCIO3, HCIO4, H 2 0 2 , Na + , CI " , H + , H “ , OH “ , 0 3 , 0 4 *” , 1 0, OH *” , HOCI-0 2 *” , HOCI-O 3 , 0 2 *” , H0 2 * , NaCI, HCI, NaOH, water clusters, or a combination thereof to a patient experiencing oxidative stress; and treating the oxidative stress related disorder.
- a composition including at least one species selected from 0 2 , H 2 , Cl 2 , OCr, HOCI, NaOCI, HCI0 2 , CI0 2 , HCIO3, HCIO4, H
- Embodiments include a method of treating a reduced mitochondrial DNA disorder comprising administering a composition including at least one species selected from 0 2 , H 2 , Cl 2 , OCI “ , HOCI, NaOCI, HCI0 2 , CI0 2 , HCIO3, HCIO4, H 2 0 2 , Na + , CI “ , H + , H “ , OH “ , 0 3 , 0 4 *” , 1 0, OH *” , HOCI-0 2 *” , HOCI-O 3 , 0 2 *” , H0 2 * , NaCI, HCI, NaOH, water clusters, or a combination thereof to a patient experiencing the reduced mitochondrial DNA disorder; increasing mitochondrial DNA density; and treating the reduced mitochondrial DNA disorder.
- disorders include sacropenia, diabetes, Alzheimer's disease, Parkinson's disease, neurological disease, muscle loss due to aging, obesity, or cardiovascular disorders.
- Figure 1 is a flow chart of a process as described herein.
- Figure 2 illustrates an example diagram of the generation of various molecules at the electrodes.
- the molecules written between the electrodes depict the initial reactants and those on the outside of the electrodes depict the molecules/ions produced at the electrodes and their electrode potentials.
- Figure 3 illustrates a plan view of a process and system for producing a composition according to the present description.
- Figure 4 illustrates an example system for preparing water for further processing into a composition described herein.
- Figure 5 illustrates a CI35 spectrum of a NaCI, NaCIO solution at a pH of 12.48, and a composition described herein (the composition is labeled "ASEA").
- Figure 6 illustrates a 1 H NMR spectrum of a composition of the present disclosure.
- Figure 7 illustrates a 31 P NMR spectrum of DIPPMPO combined with a composition described herein.
- Figure 8 illustrates a mass spectrum showing a parent peak and fragmentation pattern for DIPPMPO with m/z peaks at 264, 222, and 180.
- Figure 9 illustrates oxygen/nitrogen ratios for a composition described herein compared to water and NaCIO (the composition is labeled "ASEA").
- Figure 10 illustrates chlorine/nitrogen ratios for a composition described herein compared to water and NaCIO (the composition is labeled "ASEA").
- Figure 1 1 illustrates ozone/nitrogen ratios for a composition described herein compared to water and NaCIO (the composition is labeled "ASEA").
- Figure 12 illustrates the carbon dioxide to nitrogen ratio of a composition as described herein compared to water and NaCIO (the composition is labeled "ASEA").
- Figure 13 illustrates an EPR splitting pattern for a free electron.
- Figure 14 illustrates a flow chart of the mouse study described in Example 3 (the composition-treatment group is labeled "ASEA").
- Figure 15 is a flow chart showing a total overview of the mouse preparation and study (the composition used is referred-to as "ASEA").
- Figure 16 illustrates mice grouped into placebo and ASEA (a composition described herein) treatment versus run time and versus glycogen depletion.
- Figure 17A illustrates the fold change relate to ASEA (a composition described herein) treatment group of different mouse groups.
- Figure 17B illustrates the fold change difference between ASEA sedentary (non-running) and ASEA running groups.
- Figure 18 illustrates different mouse groups versus the amount of liver Superoxide Dismutase (SOD) produced (the composition embodiment used is referred-to as "ASEA").
- Figure 19A and 19B illustrate different mouse groups(sedentary / running; treatment / placebo) versus oxidized glutathione (the composition embodiment used is referred-to as "ASEA").
- Figure 20 illustrates different mouse groups versus fold change for IL-6 and TNF-alpha (the composition embodiment used is referred-to as "ASEA").
- Figure 21 illustrates a comparison of A-B ratios between conditions 24 hours post ingestion (the composition embodiment used is referred-to as "ASEA").
- Figure 22 illustrates a flow chart of the human running performance study protocol (the composition embodiment used in the protocol is referred-to as "ASEA").
- Figure 23 illustrates a flow chart of a 12-week, randomized trial performed accord to the protocol of Example 7 (the composition embodiment used in the protocol is referred-to as "ASEA").
- Figure 24 illustrates a graph of VC0 2 versus V0 2 resulting from the study in Example 8 (the composition embodiment used in the protocol is referred-to as "ASEA").
- Figure 25 illustrates cell images for each culture results of HMVEC-L Cells p65 subunit NF-kB screen for toxicity (the composition embodiment used in the protocol is referred-to as "ASEA").
- Figure 26 illustrates results for P-Jun screen for toxicity (the composition embodiment used in the protocol is referred-to as "ASEA").
- Figure 27 illustrates a graph showing the reduction of oxidants over an 1 1 minute interval (RFU units on vertical scale).
- Figure 28 illustrates a graph showing antioxidant activity over an 1 1 minute interval (the composition embodiment used in the protocol is referred-to as "ASEA").
- Figure 29 illustrates nuclear staining patterns for results of HMVEC-L Nuclear Accumulation of NRF2 (the composition embodiment used in the protocol is referred-to as "ASEA").
- Figure 30 illustrates serum-starved cell cultures exposed to low-concentration ASEA (a composition disclosed herein).
- Figure 31 illustrates a western blot validation of NRF2 nuclear accumulation following ASEA treatment.
- Figure 32 illustrates results for proliferation of murine and HMVEC-L cells and LDH activity following ASEA treatment.
- Figure 33 illustrates further results for proliferation of murine and HMVEC-L cells and LDH activity following ASEA treatment.
- Figure 34 illustrates results of HMVEC-L viability exposed high-concentration ASEA and to escalating amounts of Cachexin stressor (the composition embodiment used in the protocol is referred-to as "ASEA").
- Figure 35 illustrates results of concentration-dependent response of HMVEC-L cells to Cachexin insult (the composition embodiment used in the protocol is referred-to as "ASEA").
- RXNs can include, but are not limited to superoxides: 0 2 *" , H0 2 * ; hypochlorites: OCI “ , HOCI, NaOCI; hypochlorates: HCI0 2 , CI0 2 , HCIO 3 , HCI0 4 ; oxygen derivatives: 0 2 , 0 3 , 0 4 *" , 1 0; hydrogen derivatives: H 2 , H " ; hydrogen peroxide: H 2 0 2 ; hydroxyl free radicals: OH *" ; ionic compounds: Na + , CI " , H + , OH " , NaCI, HCI, NaOH; chlorine: Cl 2 ; and water clusters: n * H 2 0 - induced dipolar layers around ions.
- a composition comprising RXNs as described herein can mobilize free fatty acids (FFAs) and
- Embodiments can further comprise a PPAR agonist.
- PPARs can be nuclear receptors.
- the PPAR family comprises three isoforms, designated alpha, gamma and delta (also called beta), each encoded by a different gene. These receptors, which form part of the superfamily of nuclear receptors and of transcription factors, may play a major role in regulation of lipid and carbohydrate metabolism. PPARs may play important roles in the regulation of cellular differentiation, development, and metabolism (carbohydrate, lipid, protein), and tumorigenesis of higher organisms.
- PPAR-alpha may control lipid metabolism (hepatic and muscular) and glucose homeostasis, and may influence intracellular metabolism of lipids and sugars by direct control of transcription of the genes coding for proteins involved in lipid homeostasis. PPAR- alpha may also exert anti-inflammatory and antiproliferative effects and may prevent the proatherogenic effects of accumulation of cholesterol in macrophages by stimulating the outflow of cholesterol. PPAR-gamma is a key regulator of adipogenesis. It may also be involved in the lipid metabolism of mature adipocytes, in glucose homeostasis, in insulin resistance, in inflammation, in accumulation of cholesterol at the macrophage level and in cellular proliferation.
- PPAR-gamma consequently may play a role in the pathogenesis of obesity, insulin resistance and diabetes.
- PPAR-delta may be involved in controlling lipid and carbohydrate metabolism, in the energy balance, in neurodegeneration, in obesity, in the formation of foam cells and in inflammation.
- Methods of making therapeutic compositions comprising: electrolyzing salinated water having a salt concentration of about 10 g NaCI/gal, such as10.75 g NaCI/gal using a set of electrodes with an amperage of about 50-60 amps, such as 56 amps to form a life enhancing composition, wherein the water is chilled below room temperature and the water is circulated during electrolyzing.
- a method of producing the disclosed compositions can include one or more of the steps of (1 ) preparation of an ultra-pure homogeneous solution of sodium chloride in water, (2) temperature control and flow regulation through a set of inert catalytic electrodes and (3) a modulated electrolytic process that results in the formation of such stable molecular moieties and complexes. In one embodiment, such a process includes all these steps.
- the saline generally should be free from contaminants, both organic and inorganic, and homogeneous down to the molecular level.
- metal ions can interfere with the electro-catalytic surface reactions, and thus it may be helpful for metals to be avoided.
- a brine solution is used to salinate the water.
- the brine solution can have a NaCI concentration of about 540 g NaCI/gal, such as 537.5 g NaCI/gal.
- the composition can include at least one species such as 0 2 , H 2 , Cl 2 , OCI “ , HOCI, NaOCI, HCI0 2 , CI0 2 , HCI0 3 , HCI0 4 , H 2 0 2 , Na + , CI " , H + , H “ , OH “ , 0 3 , 0 4 *” , 1 0, OH *” , HOCI-0 2 *” , HOCI-Os, 0 2 *” , H0 2 * , NaCI, HCI, NaOH, water clusters, or a combination thereof.
- species such as 0 2 , H 2 , Cl 2 , OCI “ , HOCI, NaOCI, HCI0 2 , CI0 2 , HCI0 3 , HCI0 4 , H 2 0 2 , Na + , CI " , H + , H “ , OH “ , 0 3 , 0 4 *” ,
- the composition can include at least one species such as H 2 , Cl 2 , OCI " , HOCI, NaOCI, HCI0 2 , CI0 2 , HCIO3, HCI0 4 , H 2 0 2 , 0 3 , 0 4 *" , 1 0 2 , OH *" , HOCI- 0 2 *” , HOCI-O3, 0 2 *” , H0 2 * , water clusters, or a combination thereof.
- species such as H 2 , Cl 2 , OCI " , HOCI, NaOCI, HCI0 2 , CI0 2 , HCIO3, HCI0 4 , H 2 0 2 , 0 3 , 0 4 *" , 1 0 2 , OH *" , HOCI- 0 2 *” , HOCI-O3, 0 2 *” , H0 2 * , water clusters, or a combination thereof.
- the composition can include at least one species such as HCIO3, HCI0 4 , H 2 0 2 , 0 3 , 0 4 *" , 1 0 2 , OH *" , HOCI-0 2 *” , HOCI-O3, 0 2 *” , H0 2 * , water clusters, or a combination thereof.
- the composition can include 0 2 . In one embodiment, the composition can include H 2 . In one embodiment, the composition can include Cl 2. In one embodiment, the composition can include OCI " . In one embodiment, the composition can include HOCI. In one embodiment, the composition can include NaOCI. In one embodiment, the composition can include HCI0 2 . In one embodiment, the composition can include CI0 2 . In one embodiment, the composition can include HCI0 3 . In one embodiment, the composition can include HCI0 4 . In one embodiment, the composition can include H 2 0 2 . In one embodiment, the composition can include Na + . In one embodiment, the composition can include CI " . In one embodiment, the composition can include H + .
- the composition can include H " . In one embodiment, the composition can include OH " . In one embodiment, the composition can include 0 3 . In one embodiment, the composition can include 0 4 *" . In one embodiment, the composition can include 1 0 2 . In one embodiment, the composition can include OH *" . In one embodiment, the composition can include HOCI-0 2 *" . In one embodiment, the composition can include HOCI-0 3 , 0 2 *" . In one embodiment, the composition can include H0 2 * . In one embodiment, the composition can include NaCI. In one embodiment, the composition can include HCI. In one embodiment, the composition can include NaOH. In one embodiment, the composition can include water clusters. Embodiments can include combinations thereof.
- 100 is an optional reverse osmosis procedure 102.
- Water can be supplied from a variety of sources, including but not limited to municipal water, filtered water, nanopure water, or the like.
- the reverse osmosis process can vary, but can provide water having a total dissolved solids content of less than about 10 ppm, about 9 ppm, about 8 ppm, about 7 ppm, about 6 ppm, about 5 ppm, about 4 ppm, about 3 ppm, about 2 ppm, about 1 ppm, or the like.
- the reverse osmosis process can be performed at a temperature of about 5°C, about 10°C, about 15°C, about 20°C, about 25°C, about 30°C, about 35°C, or the like.
- the reverse osmosis step can be repeated as needed to achieve a particular total dissolved solids level.
- an optional distillation step 104 can be performed.
- the distillation process can vary, but can provide water having a total dissolved solids content of less than about 5 ppm, about 4 ppm, about 3 ppm, about 2 ppm, about 1 ppm, about 0.9 ppm, about 0.8 ppm, about 0.7 ppm, about 0.6 ppm, about 0.5 ppm, about 0.4 ppm, about 0.3 ppm, about 0.2 ppm, about 0.1 ppm, or the like.
- the temperature of the distillation process can be performed at a temperature of about 5°C, about 10°C, about 15°C, about 20°C, about 25°C, about 30°C, about 35°C, or the like.
- the distillation step can be repeated as needed to achieve a particular total dissolved solids level.
- the level of total dissolved solids in the water can be less than about 5 ppm, about 4 ppm, about 3 ppm, about 2 ppm, about 1 ppm, about 0.9 ppm, about 0.8 ppm, about 0.7 ppm, about 0.6 ppm, about 0.5 ppm, about 0.4 ppm, about 0.3 ppm, about 0.2 ppm, about 0.1 ppm, or the like.
- the reverse osmosis, distillation, both, or neither, can be preceded by a carbon filtration step.
- Purified water can be used directly with the systems and methods described herein.
- contaminants can be removed from a commercial source of water by the following procedure: water flows through an activated carbon filter to remove the aromatic and volatile contaminants and then undergoes Reverse Osmosis (RO) filtration to remove dissolved solids and most organic and inorganic contaminants.
- the resulting filtered RO water can contain less than about 8 ppm of dissolved solids.
- Most of the remaining contaminants can be removed through a distillation process, resulting in dissolved solid measurements less than 1 ppm.
- distillation may also serve to condition the water with the correct structure and Oxidation Reduction Potential (ORP) to facilitate the oxidative and reductive reaction potentials on the platinum electrodes in the subsequent electro-catalytic process.
- ORP Oxidation Reduction Potential
- a salt is added to the water in a salting step 106.
- the salt can be unrefined, refined, caked, de-caked, or the like.
- the salt is sodium chloride (NaCI).
- the salt can include an additive.
- Salt additives can include, but are not limited to potassium iodide, sodium iodidie, sodium iodate, dextrose, sodium fluoride, sodium ferrocyanide, tricalcium phosphate, calcium carbonate, magnesium carbonate, fatty acids, magnesium oxide, silicone dioxide, calcium silicate, sodium aluminosilicate, calcium aluminosilicate, ferrous fumarate, iron, or folic acid. Any of these additives can be added at this point or at any point during the described process. For example, the above additives can be added just prior to bottling.
- Salt can be added to water in the form of a brine solution.
- a physical mixing apparatus can be used or a circulation or recirculation can be used.
- pure pharmaceutical grade sodium chloride is dissolved in the prepared distilled water to form a 15 wt % sub-saturated brine solution and continuously recirculated and filtered until the salt has completely dissolved and all particles > 0.1 microns are removed. This step can take several days.
- the filtered, dissolved brine solution is then injected into tanks of distilled water in about a 1 :352 ratio (salt:water) in order to form a 0.3% saline solution.
- a ratio 10.75 g of salt per 1 gallon of water can be used to form the composition.
- 10.75 g of salt in about 3-4 g of water, such as 3.7875 g of water can be used to form the composition. This solution then can be allowed to re-circulate and diffuse until homogeneity at the molecular scale has been achieved.
- the homogenous saline solution is chilled to about 4.8 ⁇ 0.5°C. Temperature regulation during the entire electro-catalytic process can be employed as thermal energy generated from the electrolysis process itself may cause heating. In one embodiment, process temperatures at the electrodes can be constantly cooled and maintained at about 4.8°C throughout electrolysis.
- Brine can then be added to the previously treated water or to fresh untreated water to achieve a NaCI concentration of between about 1 g NaCI/gal water and about 25 g NaCI/gal water, between about 8 g NaCI/gal water and about 12 g NaCI/gal water, or between about 4 g NaCI/gal water and about 16 g NaCI/gal water.
- a NaCI concentration of between about 1 g NaCI/gal water and about 25 g NaCI/gal water, between about 8 g NaCI/gal water and about 12 g NaCI/gal water, or between about 4 g NaCI/gal water and about 16 g NaCI/gal water.
- a physical mixing apparatus can be used or a circulation or recirculation can be used.
- the salt solution can then be chilled in a chilling step 108.
- cryogenic cooling using liquid nitrogen cooling lines can be used.
- the solution can be run through propylene glycol heat exchangers to achieve the desired temperature.
- the chilling time can vary depending on the amount of liquid, the starting temperature and the desired chilled temperature.
- Products from the anodic reactions can be effectively transported to the cathode to provide the reactants necessary to form the stable complexes on the cathode surfaces. Maintaining a high degree of homogeneity in the fluids circulated between the catalytic surfaces can also be helpful.
- a constant flow of about 2-8 ml_/cm 2* sec can be used, with typical mesh electrode distances 2 cm apart in large tanks. This flow can be maintained, in part, by the convective flow of gasses released from the electrodes during electrolysis.
- the mixed solution, chilled or not, can then undergo electrochemical processing through the use of at least one electrode in an electrolyzing step 110.
- Each electrode can be or include a conductive metal. Metals can include, but are not limited to copper, aluminum, titanium, rhodium, platinum, silver, gold, iron, a combination thereof or an alloy such as steel or brass.
- the electrode can be coated or plated with a different metal such as, but not limited to aluminum, gold, platinum or silver.
- each electrode is formed of titanium and plated with platinum. The platinum surfaces on the electrodes by themselves can be optimal to catalyze the required reactions.
- Rough, double layered platinum plating can assure that local "reaction centers" (sharply pointed extrusions) are active and that the reactants not make contact with the underlying electrode titanium substrate.
- rough platinum-plated mesh electrodes in a vertical, coaxial, cylindrical geometry can be optimal, with, for example, not more than 2.5 cm, not more than 5 cm, not more than 10 cm, not more than 20 cm, or not more than 50 cm separation between the anode and cathode.
- the amperage run through each electrode can be between about 2 amps and about 15 amps, between about 4 amps and about 14 amps, at least about 2 amps, at least about 4 amps, at least about 6 amps, or any range created using any of these values. In one embodiment, 7 amps is used with each electrode.
- the amperage can be run through the electrodes for a sufficient time to electrolyze the saline solution.
- the solution can be chilled during the electrochemical process.
- the solution can also be mixed during the electrochemical process. This mixing can be performed to ensure substantially complete electrolysis.
- Electrodes can cause movement of ions. Negative ions can move toward the anode and positive ions toward the cathode. This can enable exchange of reactants and products between the electrodes. In some embodiments, no barriers are needed between the electrodes.
- an electrolyzed solution is created.
- the solution can be stored and or tested for particular properties in storage/testing step 112.
- compositions described herein can include one or more of these chemical entities, known as redox signaling agents or RXNs.
- the chlorine concentration of the electrolyzed solution can be between about 5 ppm and about 34 ppm, between about 10 ppm and about 34 ppm, or between about 15 ppm and about 34 ppm. In one embodiment, the chlorine concentration is about 32 ppm.
- the saline concentration in the electrolyzed solution can be, for example, between about 0.10% w/v and about 0.20% w/v, between about 0.1 1 % w/v and about 0.19% w/v, between about 0.12% w/v and about 0.18% w/v, between about 0.13% w/v and about 0.17% w/v, or between about 0.14% w/v and about 0.16% w/v.
- the composition generally can include electrolytic and/or catalytic products of pure saline that mimic redox signaling molecular compositions of the native salt water compounds found in and around human cells.
- the composition can be fine tuned to mimic or mirror molecular compositions of different biological media.
- the composition can have reactive species other than chlorine present.
- species present in the compositions described herein can include, but are not limited to 0 2 , H 2 , Cl 2 , OCI “ , HOCI, NaOCI, HCI0 2 , CI0 2 , HCIO3, HCIO4, H 2 0 2 , Na + , CI " , H + , H “ , OH “ , 0 3 , 0 4 *” , 1 0 2 , OH *" , HOCI- 0 2 *” , HOCI-O3, 0 2 *” , H0 2 * , NaCI, HCI, NaOH, and water clusters: n * H 2 0 - induced dipolar layers around ions, and the like.
- Compositions disclosed herein can also include PPAR agonists, such as, for example, FFAs, fibrates (fenofibrate, bezafibrate, ciprofibrate, gemfibrozil), thiazolidinediones (rosiglitazone and pioglitazone), L-165041 , GW501516, KD3010, eicosanoids, prostaglandins (E1 - alprostadil, I2 - prostacyclin, PGJ2), prostacyclins (epoprostenol, treprostinil, FLOLAN ® , veletri, remodulin, ventavis (iloprost), the thromboxanes (thromboxane A2, thromboxane B2), leukotrienes (LTC4, LTD4, LTE4 and LTF4),.
- FFAs fibrates
- fibrates fibrates
- bezafibrate fibrrofibrate,
- the PPAR agonist can be "dual", “balanced” or “pan” PPAR ligands ("glitazars"), including, for example, aleglitazar, muraglitazar, tesaglitazar, AM3102, CAY10506, CP 775146, DRF 2519, (+)-etomoxir sodium salt hydrate, GSK 3787, GW0742, GW 1929, GW 7647, GW1929 hydrate, W501516, L-165041 , methyl-8-hydroxy-8-(2-pentyl-oxyphenyl)-oct-5-ynoate, NPC 15199, nTZDpa, PAz-PC, pioglitazone, rosiglitazone (potassium salt), rosiglitazone-d3 maleate, S26948, WY 14643
- the PPAR agonist can be packaged separately from the RXN composition comprising at least one RXN.
- the PPAR agonist can be added to the RXN composition just prior to administration to a patient.
- the PPAR agonist and the RXN can be administered in separate compositions.
- the PPAR agonist can be contained within microspheres suspended within a RXN composition.
- the PPAR agonist / RXN composition can comprise a gelcap.
- Administration of PPAR agonist and/or RXN compositions can be achieved via any suitable method, including, for example, parenterally, by injection, epicutaneous, inhalational, enema, eye drops, ear drops, through mucous membranes in the body, by mouth (orally), gastric feeding tube, duodenal feeding tube, gastrostomy, rectally, intravenous, intra-arterial, intraosseous infusion, intra-muscular, intracerebral, intracerebro- ventricular, subcutaneous, or the like.
- compositions of the invention can be formulated into any suitable aspect, such as, for example, aerosols, liquids, elixirs, syrups, tinctures, creams, ointments, lotions, thin films, solids, gelcaps, a microsphere suspension, a soft gelatin capsule, and the like.
- each administration can be about 1 oz, about 2 oz, about 3 oz, about 4 oz, about 5 oz, about 6 oz, about 7 oz, about 8 oz, about 9 oz, about 10 oz, about 1 1 oz, about 12 oz, about 16 oz, about 20 oz, about 24 oz, about 28 oz, about 32 oz, about 34 oz, about 36 oz, about 38 oz, about 40 oz, about 46 oz, between about 1 oz and about 32 oz, between about 1 oz and about 16 oz, between about 1 oz and about 8 oz, at least about 2 oz, at least about 4 oz, or at least about 8 oz.
- the composition can be administered at a rate of about 4 oz twice a day
- the administration can be acute or long term.
- the composition can be administered for a day, a week, a month, a year or longer.
- compositions described herein when administered can be used to treat a condition or a disease.
- the compositions described herein can increase the density of mitochondrial DNA.
- an increase in mitochondrial DNA of about 1 %, about 5%, about 10%, about 15%, about 20%, about 21 %, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 32%, about 34%, about 36%, about 38%, about 40%, about 45%, between about 1 % and about 40%, between about 1 % and about 10%, between about 20% and about 30%, at least about 5%, at least about 10%, or at least about 20% when compared to an individual who has not taken the composition.
- An increase in mitochondrial DNA can result in a lower level of free radicals in the blood which can in turn lead to a reduced amount of oxidative stress.
- compositions disclosed herein can be useful in treating diseases related to mitochondrial DNA.
- the compositions described can treat conditions or diseases such as, but not limited to sacropenia, Parkinson's disease, neuro-related age disease, obesity, aging, life stresses such as those caused by fear, neurodegenerative diseases, cognitive disorders, obesity, reduced metabolic rate, metabolic syndrome, diabetes mellitus, cardiovascular disease, hyperlipidemia, neurodegenerative disease, cognitive disorder, mood disorder, stress, and anxiety disorder; for weight management, or to increase muscle performance or mental performance, AIDS, dementia complex, Alzheimer's disease, amyotrophic lateral sclerosis, adrenoleukodystrophy, Alexander disease, Alper's disease, ataxia telangiectasia, Batten disease, bovine spongiform encephalopathy (BSE), Canavan disease, corticobasal degeneration, Creutzfeldt-Jakob disease, dementia with Lewy bodies, fatal familial insomnia, frontotemporal lobar degeneration, Huntington's disease, Kennedy
- compositions disclosed herein can be useful in treating diseases involving PPAR pathways, including, for example, metabolic syndrome, cardiovascular disease, diabetes, obesity, glucose intolerance, hyperinsulinemia, hypercholesterolemia, hypertriglyceridemia, and hypertension, inflammation, vascular function, and vascular remodeling, cancer, inflammation, neurodegenerative diseases, diseases relating to mitochondrial biogenesis, ageing, and the like.
- diseases involving PPAR pathways including, for example, metabolic syndrome, cardiovascular disease, diabetes, obesity, glucose intolerance, hyperinsulinemia, hypercholesterolemia, hypertriglyceridemia, and hypertension, inflammation, vascular function, and vascular remodeling, cancer, inflammation, neurodegenerative diseases, diseases relating to mitochondrial biogenesis, ageing, and the like.
- compositions of the invention can comprise a component of a therapy acting through multiple mechanisms.
- a dual acting therapy can comprise a first therapy which activates a PPAR pathway and at least one other agent that does not activate a PPAR pathway.
- the first agent can be at least one RXN.
- the at least one other agent is a mAb, radiation, surgery, angiogenesis inhibitor, transplantation, a cancer vaccine, gene therapy, laser treatment, photodynamic therapy, an alkylating agent, an antimetabolite, an anti-tumor antibiotic, a topoisomerase inhibitor, a mitotic inhibitor, a corticosteroid, hormone therapy, immunotherapy, and the like.
- Embodiments can provide a substrate in vitro for superoxides and/or fatty acid oxidation (FAO) enzymes comprising RXNs.
- Embodiments can include a method of treating cancer comprising a combination therapy wherein one therapy comprises compositions comprising at least one RXN which increases FFAs, and at least one other agent wherein the at least one other agent does not increase FFAs.
- methods of treating an oxidative stress related disorders comprising: administering a composition including at least one species selected from 0 2 , H 2 , Cl 2 , OCI “ , HOCI, NaOCI, HCI0 2 , CI0 2 , HCI0 3 , HCI0 4 , H 2 0 2 , Na + , CI " , H + , H “ , OH “ , 0 3 , 0 4 *” , 1 0, OH *” , HOCI-0 2 *” , HOCI-O3, 0 2 *” , H0 2 * , NaCI, HCI, NaOH, water clusters, or a combination thereof to a patient experiencing oxidative stress; and treating the oxidative stress related disorder.
- the administration occurs twice a day or once a day. Each administration can include between about 1 oz and about 16 oz per day.
- the oxidative stress related disorder is diabetes, cardiovascular
- Figure 3 illustrates a plan view of a process and system for producing a life enhancing composition according to the present description.
- One skilled in the art understands that changes can be made to the system to alter the life enhancing composition, and these changes are within the scope of the present description.
- Incoming water 202 can be subjected to reverse osmosis system 204 at a temperature of about 15-20°C to achieve purified water 206 with about 8 ppm of total dissolved solids. Purified water 206, is then fed at a temperature of about 15-20°C into distiller 208 and processed to achieve distilled water 210 with about 0.5 ppm of total dissolved solids. Distilled water 210 can then be stored in tank 212.
- FIG. 4 illustrates an example system for preparing water for further processing into a therapeutic beverage.
- System 300 can include a water source 302 which can feed directly into a carbon filter 304. After oils, alcohols, and other volatile chemical residuals and particulates are removed by carbon filter 304, the water can be directed to resin beds within a water softener 306 which can remove dissolved minerals. Then, as described above, the water can pass through reverse osmosis system 204 and distiller 208.
- distilled water 210 can be gravity fed from tank 212 into saline storage tank cluster 214 using line 216.
- Saline storage tank cluster 214 in one embodiment can include twelve tanks 218. Each tank 218 can be filled to about 1 ,300 gallons with distilled water 210.
- a handheld meter can be used to test distilled water 210 for salinity.
- Saline storage tank cluster 214 is then salted using a brine system 220.
- Brine system 220 can include two brine tanks 222. Each tank can have a capacity of about 500 gallons. Brine tanks 222 are filled to 475 gallons with distilled water 210 using line 224 and then NaCI is added to the brine tanks 222 at a ratio of about 537.5 g/gal of liquid. At this point, the water is circulated 226 in the brine tanks 222 at a rate of about 2,000 gal/hr for about 4 days.
- the salinity of the water in tanks 218 can be tested using a handheld conductivity meter such as an YSI ECOSENSE® ecp300 (YSI Inc., Yellow Springs, OH). Any corrections based on the salinity measurements can be made at this point.
- Brine solution 228 is then added to tanks 218 to achieve a salt concentration of about 10.75 g/gal.
- the salted water is circulated 230 in tanks 218 at a rate of about 2,000 gal/hr for no less than about 72 hours. This circulation is performed at room temperature.
- a handheld probe can again be used to test salinity of the salinated solution. In one embodiment, the salinity is about 2.8 ppth.
- the amount of liquid remaining in the tanks is measured.
- the amount of liquid remaining in a tank is measured by recording the height that the liquid level is from the floor that sustains the tank, in centimeters, and referencing the number of gallons this height represents. This can be done from the outside of the tank if the tank is semi-transparent.
- the initial liquid height in both tied tanks can also be measured.
- distilled water can be pumped in.
- the amount of distilled water that is being pumped into a holding tank can then be calculated by measuring the rise in liquid level: subtracting the initial height from the filled height and then multiplying this difference by a known factor.
- the amount of salt to be added to the tank is then calculated by multiplying 1 1 grams of salt for every Gallon of distilled water that has been added to the tank. The salt can be carefully weighed out and dumped into the tank.
- the tank is then agitated by turning on the recirculation pump and then opening the top and bottom valves on the tank. Liquid is pumped from the bottom of the tank to the top.
- the tank can be agitated for three days before it may be ready to be processed.
- the salinity is checked with a salinity meter by taking a sample from the tank and testing it. Salt or water can be added to adjust the salinity within the tanks. If either more water or more salt is added then the tanks are agitated for 6 more hours and tested again. After about three days of agitation, the tank is ready to be processed.
- Salinated water 232 is then transferred to cold saline tanks 234. In one embodiment, four 250 gal tanks are used. The amount of salinated water 232 moved is about 1 ,000 gal. A chiller 236 such as a 16 ton chiller is used to cool heat exchangers 238 to about 0-5°C. The salinated water is circulated 240 through the heat exchangers which are circulated with propylene glycol until the temperature of the salinated water is about 4.5- 5.8°C. Chilling the 1 ,000 gal of salinated water generally takes about 6-8 hr.
- a chiller 236 such as a 16 ton chiller is used to cool heat exchangers 238 to about 0-5°C.
- the salinated water is circulated 240 through the heat exchangers which are circulated with propylene glycol until the temperature of the salinated water is about 4.5- 5.8°C. Chilling the 1 ,000 gal of salinated water generally takes about 6-8 hr.
- Cold salinated water 242 is then transferred to processing tanks 244.
- processing tanks 244 In one embodiment, eight tanks are used and each can have a capacity of about 180 gal.
- Each processing tank 244 is filled to about 125 gal for a total of 1 ,000 gal.
- Heat exchangers 246 are again used to chill the cold salinated water 242 added to processing tanks 244.
- Each processing tank can include a cylinder of chilling tubes and propylene glycol can be circulated.
- the heat exchangers can be powered by a 4-5 ton chiller 248.
- the temperature of cold salinated water 242 can remain at 4.5-5.8°C during processing.
- the aged salt water Prior to transferring aged salt water to processing tanks, the aged salt water can be agitated for about 30 minutes to sufficiently mix the aged salt water. Then, the recirculation valves can then be closed, the appropriate inlet valve on the production tank is opened, and the tank filled so that the salt water covers the cooling coils and comes up to the fill mark (approximately 125 gallons).
- the pump is turned off but the chiller left on.
- the tank should be adequately agitated or re-circulated during the whole duration of electrochemical processing and the temperature should remain constant throughout.
- Each processing tank 244 includes electrode 250. Electrodes 250 can be 3 inches tall circular structures formed of titanium and plated with platinum. Electrochemical processing of the cold salinated water can be run for 8 hr. A power supply 252 is used to power the eight electrodes (one in each processing tank 244) to 7 amps each for a total of 56 amps. The cold salinated water is circulated 254 during electrochemical processing at a rate of about 1 ,000 gal/hr.
- An independent current meter can be used to set the current to around 7.0 Amps. Attention can be paid to ensure that the voltage does not exceed 12V and does not go lower than 9V. Normal operation can be about 10V.
- a run timer can be set for a prescribed time (about 4.5 to 5 hours).
- Each production tank can have its own timer and/or power supply. Electrodes should be turned off after the timer has expired.
- the production tanks can be checked periodically.
- the temperature and/or electrical current can be kept substantially constant.
- the electrodes can be visible from the top, emitting visible bubbles.
- small bubbles of un- dissolved oxygen can start building up in the tank as oxygen saturation occurs, obscuring the view of the electrodes.
- a slight chlorine smell can be normal.
- life enhancing water 256 has been created with a pH of about 6.8-8.2, 32 ppm of chlorine, 100% OCI " and 100% O "2 .
- the composition 256 is transferred to storage tanks 258.
- a composition produced as described in Example 1 and marketed under the trade name ASEA® was analyzed using a variety of different characterization techniques. ICP/MS and 35 CI NMR were used to analyze and quantify chlorine content. Headspace mass spectrometry analysis was used to analyze adsorbed gas content in the beverage. 1 H NMR was used to verify the organic matter content in the beverage. 31 P NMR and EPR experiments utilizing spin trap molecules were used to explore the beverage for free radicals.
- Sodium hypochlorite solutions were prepared at different pH values. 5% sodium hypochlorite solution had a pH of 12.48. Concentrated nitric acid was added to 5% sodium hypochlorite solution to create solutions that were at pH of 9.99, 6.99, 5.32, and 3.28. These solutions were then analyzed by NMR spectroscopy. The beverage had a measured pH of 8.01 and was analyzed directly by NMR with no dilutions.
- NMR spectroscopy experiments were performed using a 400 MHz Bruker spectrometer equipped with a BBO probe. 35 CI NMR experiments were performed at a frequency of 39.2 MHz using single pulse experiments. A recycle delay of 10 seconds was used, and 128 scans were acquired per sample. A solution of NaCI in water was used as an external chemical shift reference. All experiments were performed at room temperature.
- An ASEA sample was prepared by adding 550 ⁇ _ of ASEA and 50 ⁇ _ of D 2 0 (Cambridge Isotope Laboratories) to an NMR tube and vortexing the sample for 10 seconds.
- 1 H NMR experiments were performed on a 700 MHz Bruker spectrometer equipped with a QNP cryogenically cooled probe. Experiments used a single pulse with pre-saturation on the water resonance experiment. A total of 1024 scans were taken. All experiments were performed at room temperature.
- a 1 H NMR spectrum of the composition was determined and is presented in Figure 6. Only peaks associated with water were able to be distinguished from this spectrum. This spectrum show that very little if any organic material can be detected in the composition using this method.
- DIPPMPO (5-(Diisopropoxyphosphoryl)-5-1-pyrroline-N-oxide) (VWR) samples were prepared by measuring about 5 mg of DIPPMPO into a 2 mL centrifuge tube. This tube then had 550 ⁇ _ of either the composition or water added to it, followed by 50 ⁇ _ of D 2 0. A solution was also prepared with the composition but without DIPPMPO. These solutions were vortexed and transferred to NMR tubes for analysis. Samples for mass spectrometry analysis were prepared by dissolving about 5 mg of DIPPMPO in 600 ⁇ _ of the composition and vortexing, then diluting the sample by adding 100 ⁇ _ of sample and 900 ⁇ _ of water to a vial and vortexing.
- NMR experiments were performed using a 700 MHz Bruker spectrometer equipped with a QNP cryogenically cooled probe. Experiments performed were a single 30° pulse at a 31 P frequency of 283.4 MHz. A recycle delay of 2.5 seconds and 16384 scans were used. Phosphoric acid was used as an external standard. All experiments were performed at room temperature.
- Mass spectrometry experiments were performed by directly injecting the ASEA DIPPMPO sample into a Waters/Synapt Time of Flight mass spectrometer. The sample was directly injected into the mass spectrometer, bypassing the LC, and monitored in both positive and negative ion mode.
- FIG. 7 illustrates a 31 P NMR spectrum of DIPPMPO combined with the composition.
- the peak at 21.8 ppm was determined to be DIPPMPO and is seen in both the spectrum of DIPPMPO with the composition ( Figure 7) and without the composition (not pictured).
- the peak at 24.9 ppm is most probably DIPPMPO/ ⁇ as determined in other DIPPMPO studies. This peak may be seen in DIPPMPO mixtures both with and without the composition, but is detected at a much greater concentration in the solution with the composition.
- Mass spectral data was collected in an attempt to determine the composition of the unidentified radical species.
- the mass spectrum shows a parent peak and fragmentation pattern for DIPPMPO with m/z peaks at 264, 222, and 180, as seen in Figure 8.
- Figure 8 also shows peaks for the DIPPMPO/Na adduct and subsequent fragments at 286, 244, and 202 m/z.
- Figure 8 demonstrates peaks for one DIPPMPO/radical complex with m/z of 329.
- the negative ion mode mass spectrum also had a corresponding peak at m/z of 327. There are additional peaks at 349, 367, and 302 at a lower intensity as presented in Figure 8. None of these peaks could be positively confirmed.
- the peak generated at 329 could be a structure formed from a radical combining with DIPPMPO. Possibilities of this radical species include a nitroxyl-peroxide radical ( ⁇ - ⁇ ) that may have formed in the beverage as a result of reaction with nitrogen from the air.
- Another peak at 349 could also be a result of a DIPPMPO/radical combination.
- a possibility for the radical may be hypochlorite-peroxide ( ⁇ ).
- the small intensity of this peak and small intensity of the corresponding peak of 347 in the negative ion mode mass spectrum indicate this could be a very low concentration impurity and not a compound present in the ASEA composition.
- the sonicator was set to degas which allowed for any dissolved gasses to be released from the sample into the headspace. After degassing, the samples were placed on a CTC PAL autosampler equipped with a heated agitator and headspace syringe. The agitator was set to 750 rpm and 95°C and the syringe was set to 75°C. Each vial was placed in the agitator for 20 min prior to injection into the instrument. A headspace volume of 2.5 mL was collected from the vial and injected into the instrument.
- the instrument used was an Agilent 7890A GC system coupled to an Agilent 5975C EI/CI single quadrupole mass selective detector (MSD) set up for electron ionization.
- MSD mass selective detector
- the GC oven was set to 40°C with the front inlet and the transfer lines being set to 150°C and 155°C respectively.
- the carrier gas used was helium and it was set to a pressure of 15 PSI.
- the MSD was set to single ion mode (SIM) in order to detect the following analytes:
- the ionization source temperature was set to 230°C and the quadrupole temperature was set to 150°C.
- the electron energy was set to 15 V.
- Mass spectrometry data was obtained from analysis of the gas phase headspace of the water, the composition, and hypochlorite solution.
- the raw area counts obtained from the mass spectrometer were normalized to the area counts of nitrogen in order to eliminate any systematic instrument variation.
- Both nitrogen and water were used as standards because they were present in equal volumes in the vial with nitrogen occupying the headspace and water being the solvent. It was assumed that the overall volume of water and nitrogen would be the same for each sample after degassing. In order for this assumption to be correct, the ratio of nitrogen to water should be the same for each sample.
- a cutoff value for the percent relative standard deviation (% RSD) of 5% was used. Across all nine samples, a % RSD of 4.2 was observed. Of note, sample NaCIO "3 appears to be an outlier, thus, when removed, the % RSD drops to 3.4%.
- Figures 9-1 1 illustrate oxygen/nitrogen, chlorine/nitrogen, and ozone/nitrogen ratios. It appears that there were less of these gases released from the composition than from either water or nitrogen. It should be noted that the signals for both ozone and chlorine were very weak. Thus, there is a possibility that these signals may be due to instrument noise and not from the target analytes.
- Figure 12 illustrates the carbon dioxide to nitrogen ratio. It appears that there may have been more carbon dioxide released from the composition than oxygen. However, it is possible that this may be due to background contamination from the atmosphere.
- composition samples Two different composition samples were prepared for EPR analysis. The composition with nothing added was one sample. The other sample was prepared by adding 31 mg of DIPPMPO to 20 ml. of the composition (5.9mM), vortexing, and placing the sample in a 4°C refrigerator overnight. Both samples were placed in a small capillary tube which was then inserted into a normal 5 mm EPR tube for analysis.
- EPR experiments were performed on a Bruker EMX 10/12 EPR spectrometer. EPR experiments were performed at 9.8 GHz with a centerfield position of 3500 Gauss and a sweepwidth of 100 Gauss. A 20 mW energy pulse was used with modulation frequency of 100 kHz and modulation amplitude of 1 G. Experiments used 100 scans. All experiments were performed at room temperature.
- FIG. 9 shows the EPR spectrum generated from DIPPMPO mixed with the composition.
- the composition alone showed no EPR signal after 100 scans (not presented).
- Figure 13 illustrates an EPR splitting pattern for a free electron. This electron appears to be split by three different nuclei. The data indicate that this is a characteristic splitting pattern of 0 ⁇ radical interacting with DMPO (similar to DIPPMPO). This pattern can be described by 14 N splitting the peak into three equal peaks and 1 H three bonds away splitting that pattern into two equal triplets. If these splittings are the same, it leads to a quartet splitting where the two middle peaks are twice as large as the outer peaks.
- This pattern may be seen in Figure 13 twice, with the larger peaks at 3457 and 3471 for one quartet and 3504 and 3518 for the other quartet.
- the 14 N splitting and the 1 H splitting are both roughly 14G, similar to an ⁇ radical attaching to DMPO.
- the two quartet patterns in Figure 13 are created by an additional splitting of 47G. This splitting is most likely from coupling to 31 P, and similar patterns have been seen previously.
- the EPR spectrum in Figure 13 indicates that there is a DIPPMPO/ ⁇ radical species in the solution.
- ASEA or placebo (same ingredients as ASEA composition without the proprietary signaling molecules added) was administered to the mice via gavage once per day for 1 -week.
- mice were not palatable and the mice did not drink it willingly. Gavage was an acceptable alternative to ensure the mice did not become dehydrated simply because they would not drink the study composition. The gavaging was performed by the animal husbandry staff at CLAS. In one embodiment, mice can be given an amount of the composition that is equivalent to a daily human dose as described herein.
- mice were euthanized and tissues harvested for further analysis of outcome measures.
- the four groups of mice were phased into the 1-week protocol each day. For example, if Group 1 started the protocol on a given day, Group 2 would begin the protocol on the following day, Group 3 would be begin the following day, and Group 4 the day after that. Mice from Group 1 would then be euthanized following the final treadmill test (7th day of treatment), Group 2, Group 3, and Group 4 each on subsequent days. Thus, total time for the mouse protocol was 1 1 days. There was overlap of orientation treadmill days, with maximal treadmill testing and euthanasia days. As stated, prior to euthanasia, mice from Group 1 and Group 3 underwent an endurance treadmill test to exhaustion using the protocol summarized in the following Table.
- mice were oriented (trained) to the treadmill for 15 min/day. Speeds for the training days were about 10 m/min, 15 m/min, and 18 m/min respectively. Then, on the final day of treatment mice underwent a maximal endurance capacity test on the treadmill.
- mice from Group 2 and Group 4 were not submitted to an endurance capacity test and were euthanized at the end of 1 -week treatment. Tissues harvested from these mice were collected to assess the chronic effects of the test composition in absence of an exercise intervention. All blood/plasma and tissues were snap-frozen in liquid nitrogen and stored at -80°C until assayed.
- mice were run on a multi- lane rodent treadmill (Columbus Instruments, Columbus OH) equipped with a shock grid at the back. Once each mouse was placed in a treadmill lane, a 1 minute resting period was initiated. At this point, the mouse was able to adjust to the inside of the treadmill chamber. Following the 1 minute rest period, the treadmill belt was started at a speed of about 10 m/min, and the protocol described in the above Table was followed.
- mice were allowed to run until they were no longer able to keep up with the belt and the hind limbs stayed on the shock grid for more than about 5 seconds. When the mouse was no longer running (as assessed by sitting on the shock grid with all 4 paws off of the belt for more than 5 seconds), the mouse was removed from the shock grid immediately and placed back into the home cage. The mice were then monitored for recovery for a period of at least about 20 minutes following the orientation bouts.
- the signs of exhaustion used included a mouse sitting on the shock grid for more than 5 seconds, rapid breathing, and/or increased heart rate. It has been our experience that mice that are not fatigued do not show these signs and will continue to run within 5 seconds of stopping. These procedures follow national recommendations (American Physiological Society's, Resource Book for the Design of Animal Exercise Protocols, 2006) based on research in the area. If at any point during the test a mouse got its foot caught between the shock grid and the treadmill the test was immediately terminated. If the mouse was injured and needed treatment, proper procedures were followed and vivarium staff was notified. If the mouse was deemed not injured, it was allowed to recover and placed back in its home cage and re-tested the following day.
- mice were placed back into its home cage.
- mice are usually back up and jumping around the cage within 30 seconds of re-exposure to the home cage following an endurance test.
- mice were still monitored several times during the 20-60 minutes following the procedure and notes were taken of any abnormalities such as apathy or decreased food consumption.
- Some form of motivation was needed to make the mice run on the treadmill, particularly in the orientation sessions.
- a variety of forms of motivation can be used.
- the three most common techniques are, use of shock grid, use of air puffs, and manually tapping a mouse's tail.
- Use of air puffs have the potential to be ineffective and possibly confounding to data analysis.
- manually tapping the tail was not ideal.
- shock grids were the best method of motivation for exercise on the treadmill.
- the shock grid was positioned at the back of the treadmill.
- the shock grid delivered pulsed shock at an average current of 1.0 milliamperes at 150 volts (the shock grid was adjustable within a range of 0-3.4 mA).
- the shock grid was regularly checked with an ampmeter to ensure proper functioning.
- the shock levels used were 22 times less than that accepted in the literature.
- the amperage of the system was 167-500 times less than lethal levels for mice, and the total power of the system was 60 times less than lethal levels for mice. No new data or guidelines existed to suggest that the use of a shock grid with our proposed settings was anything but appropriate.
- Figure 16 illustrates that mice who were administered ASEA had an increased run time to exhaustion. As such, ASEA can be used to increase time to exhaustion in athletes when exercising.
- ASEA consumption in sedentary mice did not increase muscle mitochondria density.
- An interaction between one long endurance exercise bout to exhaustion was observed with ASEA vs.
- ASEA sedentary P ⁇ 0.05. Fold change increased when ASEA was delivered along with exercise, but fell when exercise was not present. This supports that ASEA helped decrease the level of oxidative stress in the muscle.
- Figure 18 illustrates that SOD produced in the liver decreases in mice when administered ASEA and subjected to exercise.
- U is the amount of enyzme needed to inhibit 50% dismutation of the superoxide radical.
- An acute bout of exercise activates CuZnSOD activity, but most studies reported no change in its mRNA and enzyme protein levels, suggesting that the increased activity was due to increased O 2- concentration. This result can indicate that ASEA linked to exercise can reduce oxidative stress.
- Figures 19A and 19B illustrate that oxidized glutothione decreases in mice when administered ASEA and subjected to exercise. This result can indicate that ASEA linked to exercise can reduce oxidative stress.
- Figure 20 illustrates that exercise increased mRNA (gene expression) for IL-6 and TNF-alpha, indicating the typical pro-inflammatory response.
- ASEA tended to reduce gene expression for these inflammatory cytokines.
- Athletes ingesting ASEA for seven days started the 75 km cycling trial with high blood free fatty acids leading to increased fat oxidation and a sparing of amino acids (and potentially muscle glycogen).
- Metabolites and glycogen utilization are altered when a composition of the invention is used alongside exercise.
- a 12-Week, randomized trial is performed accord to the protocol in Figure 23.
- the study evaluates the effectiveness of 4 fl oz/day ASEA compared to placebo over a 12- week period in helping adult women improve disease risk factors associated with arterial stiffness, inflammation, cholesterol status, blood pressure, oxidative stress and capacity, fasting serum glucose, and metabolic hormones.
- Ingestion of ASEA over a 12 week period decreases arterial stiffness, decreases inflammation, improves cholesterol status, decreases blood pressure, decreases oxidative stress and capacity, decreases fasting serum glucose, and alters metabolic hormones.
- VT Ventilatory Threshold
- the immune-supporting supplement a composition of the invention, contains a balanced mixture of Redox Signaling molecules that purportedly increases the efficiency of the communication channels between cells, enabling faster response of the immune system and cellular healing activities. Enzymes in the body also break down these Redox Signaling molecules into salt water and nascent oxygen.
- Redox Signaling There are two proposed mechanisms involving Redox Signaling that can affect athletic performance, (1 ) increased efficiencies in cellular absorption or use of oxygen, prolonging aerobic metabolism, and (2) more efficient processing of lactate energy stores and tissue repair mechanisms, prolonging anaerobic metabolism.
- Anaerobic metabolism supplies the excessive demand for energy but is accompanied by the production of C0 2 and lactates. Prolonged or excessive anaerobic metabolism depletes the available energy stores faster than they can be renewed; the buildup of C0 2 and lactates can also interfere with aerobic metabolism and thus, when the energy stores are spent, exhaustion will result.
- VT Ventilatory Threshold
- VT was determined graphically from the VC0 2 vs. V0 2 graph.
- VC0 2 is the volume of C0 2 expired per minute and V0 2 is the volume of 02 inspired per minute.
- V0 2ma x is simply the maximum volume of 0 2 inspired per minute possible for any given individual.
- V0 2ma x is measured in mL/kg/min (milliliters of 0 2 per kilogram of body weight per minute).
- V0 2ma x is measured at the peak of the V0 2 curve.
- the Aerobic Threshold (AeT) was determined by the software and indicates when fat-burning metabolic activities start to be dominated by aerobic metabolism.
- the Anaerobic Threshold (AT) was also software-determined and marks the point where the anaerobic metabolism starts to completely dominate.
- V0 2max testing was done at an athletic club by accredited professionals holding degrees in exercise physiology and with more than 10 years daily experience in administering V0 2 tests. The participants were given a choice of performing the test on either a treadmill or a stationary cycle.
- a CARDIOCOACH® metabolic cart measured heart rate (HR), inspired and expired gases (V0 2 , VC0 2 , VE) and recorded weight, height, age, and body mass indexes (BMI). Power settings on the treadmill or cycle were recorded every minute.
- Each participant was scheduled to take two V0 2m ax tests, (1 ) a baseline test and (2) a final test.
- the baseline test was performed before any supplement ingestion.
- the participants drank 4 oz. of the supplement per day between the baseline test and the final test (7 to 10 days later) and drank 8 oz. of the supplement ten minutes before starting the final test.
- the power settings on the cycle or treadmill were determined by the test administrator.
- the power settings for the final test were matched exactly to the power settings of the baseline test for each participant. Participants were encouraged to strictly maintain their regular diet and exercise routine and to come to each test well hydrated (at least 8 oz. of water in the last 2 hours before each test).
- Each participant was fitted with a breathing mask and heart monitor.
- Each 0 2 max test consisted of a 10 minute warm up period where participants walked or cycled at a low power setting determined by the administrator. This was followed by a ramp up period, where the administrators increased the power settings every minute, according to their evaluation of the physical condition of the participant, and termination when the administrators started seeing the indications of a maximum V0 2 reading when RER (VC0 2 /V0 2 ) > 1.0 or at the administrator's discretion.
- the administrators had ample experience in obtaining consistent V0 2max results on this equipment, estimated at about 6% test to test variation over the last 5 years.
- the raw data (HR, V0 2 , VC0 2 , VE, Power Settings) were collected from the CARDIOCOACH® software for analysis. Data points were automatically averaged over 15 to 25 second breath intervals by the software, V0 2max is also determined by the software with an averaged V0 2 peak method. VT was determined graphically from the slope of the VC0 2 vs. V0 2 graph.
- V0 2m ax reading over all participants was measured at the relatively high value of 62.5 mL/kg/min, indicative of the quality of athletes in the sample. Only four participants had V0 2m ax readings below 55 mL/kg/min; these four were not involved in competitive training programs.
- Specific investigations include in vitro toxicity and antioxidant efficiencies of the master antioxidants glutathione peroxidase (GPx) and Superoxide Dismutase (SOD) inside living cells and the translocation of two well-studied transcription factors (NF-kB, NRF2) known to regulate toxic response and antioxidant production in human cells.
- the objectives of the investigations were (1 ) to determine if any signs of toxicity (NF-kB activation) are manifest when varying concentrations of a certain redox signaling compound, ASEA, are placed in physical contact with living cells, (2) to determine if such direct contact affects the antioxidant efficacy of glutathione peroxidase (GPx) and superoxide dismutase (SOD) and (3) to determine if such contact activates translocational transcription (NRF2) associated with increased expression of antioxidants in living human endothelial cells and to verify the expression of such transcription factors by Western Blot analysis, (4) to determine the effect of this redox signaling compound on proliferation cell counts of human cells and associated markers (LDH) for cell viability and health, (5) to determine the effects of this redox signaling compound on cells that were stressed with cytokines (Cachexin), radiation and serum starvation.
- ASEA a certain redox signaling compound
- the immune-supporting composition contains a red ox-balanced mixture of RXN [both reactive oxygen species (ROS) and reduced species (RS)] that are involved in a large variety of pathways and receptor-site activity in human cells.
- RXN reactive oxygen species
- RS reduced species
- redox signaling molecules when unbalanced or isolated, to elicit immediate recognizable toxic responses in exposed living cells; hydrogen peroxide is one example of such a redox signaling molecule.
- the first-line cellular response to toxic substances involves the translocation of NF-kB into the nucleus as a precursor to the inflammatory response and other defense mechanisms.
- the movement of NF-kB into the nucleus can be visibly tracked in a living cell under a fluorescence microscope with the aid of fluorescent tag molecules.
- the observation of nuclear translocation of NF-kB is a sure marker that a toxic response has been initiated.
- NRF2 nuclear translocation of NRF2 can be seen in cells under a fluorescence microscope. NRF2 nuclear translocation is a second-line-of-defense mechanism known to increase the production of protective enzymes and antioxidants such as glutathione peroxidase and superoxide dismutase.
- NRF2 translocation will often accompany low-level NF-kB activation and NF-kB activation (almost) always precedes NRF2 translocation.
- Enzymatic efficacy of antioxidants can be determined through standardized ELISA tests that measure the time-related reduction of certain oxidants introduced into cell lysates after the living cells have been exposed to the test substance for a given period of time.
- the reagents of the ELISA test must be chosen as not to interfere or interact with the test substance. Other critical factors such as the time of exposure and concentration dependence must be experimentally determined.
- Cachexin is a potent toxin, a cytokine, that elicits immediate toxic responses and build-up of oxidative stress in exposed cells. Cells, so stressed, exhibit a greater tendency to undergo apoptosis and die, thereby releasing internal proteins (such as LDH) into the surrounding serum.
- Glutathione peroxidase (GPx) and superoxide dismutase (SOD) ELISAs were used to determine whether a composition of the invention alters enzymatic activity in murine epidermal (JB6) cells.
- LDH (non-specific cellular death) levels and cell proliferation rates were determined for various cell types exposed to a composition of the invention.
- HMVEC-L Human microvascular endothelial lung cells
- HMVEC-L cells were treated with a phosphate buffered saline solution (PBS)negative control, 5% and 20% concentrations of a composition of the invention and a Cachexin positive control to determine the nuclear translocation activity of the p65 subunit of NF-kB (cytokine transcription) at 30, 60, 90 and 120 min intervals. Fluorescent microscopy techniques were employed to image cellular response.
- PBS phosphate buffered saline solution
- Step (4) was repeated except nuclear translocation activity of P-Jun was determined as an extension/verification of step 4.
- step (4) was repeated except nuclear translocation activity of P-Jun was determined as an extension/verification of step 4.
- [00218] Two cultures of HMVEC-L cells, one with normal random cell cycles and another with serum starvation were treated with low ⁇ 1 % concentrations of a composition of the invention to determine the nuclear activity of NRF2 (antioxidant transcription) at 30, 60, 90 and 120 minute intervals compared to a negative (PBS) control.
- NRF2 antioxidant transcription
- HMVEC-L cells (catalog* CC-2527) were purchased from Lonza (Walkersville, MD) as cryopreserved cells (Lot# 7F4273). Cells were thawed and maintained according to manufacturer's directions.
- Cell culture medium (proprietary formulation provided by Lonza) contained epidermal growth factor, hydrocortisone, GA-1000, fetal bovine serum, vasoactive endothelial growth factor, basic fibroblast growth factor, insulin growth factor-1 and ascorbic acid.
- HMVEC-L Cell cultures in normal random cell cycles were exposed to high- concentration ASEA in the serum medium, concentrations of 5% and 20%, and analyzed in conjunction with cultures exposed to phosphate buffered saline solution (PBS) as non-toxic negative control and Cachexin (5 ng/mL) as a positive control (highly toxic).
- PBS phosphate buffered saline solution
- Cachexin 5 ng/mL
- Results of HMVEC-L Cells p65 subunit NF-kB screen for toxicity Typical cell images are shown below for each culture. Translocation of p65 subunit of NF-kB into the nucleus was not seen in any cell cultures exposed to high-concentration a composition of the invention. Automated analysis confirmed this and indicated no toxic response at 0, 30, 90 and 120 minutes. In contrast, Cachexin exposed cells exhibited an immediate sustained toxic response (Figure 25).
- Cachexin is positive control and induces the translocation of p65 subunit of NF- kB from cytosol into nucleus.
- DAPI staining shows position of nuclei in these images (see white arrow).
- a composition of the invention (5 and 20% final v/v) did not induce nuclear translocation of NF-kB at 30, 60 and 120 min time points.
- Results for P-Jun screen for toxicity ( Figure 26): AP-1 index determined using anti-phospho-Jun (P-Jun) antibody.
- AP-1 is nuclear localized and upon activation, the phosphorylation status of P-Jun is increased.
- Anti-P-Jun antibody binds to the phosphorylated form reflected as an increase in fluorescence intensity (see Cachexin control).
- a consistent trend reflecting an increase in P-Jun levels was not observed for cells treated with 5% or 20% ASEA at 30, 60 and 120 min time points, while the Cachexin positive control significantly increased nuclear P-Jun levels at 30 min.
- NF-kB and P-Jun are typically the first responders to serum toxicity and are known to initiate the inflammatory response, especially in the ultrasensitive human endothelial cells, healthy human cells when directly exposed to a composition of the invention, are not expected to exhibit defensive behavior nor initiate inflammatory processes (such as the release of inflammatory cytokines). It is not certain from this data whether exposure would suppress or reverse the inflammatory process.
- Blood serum levels of such redox signaling molecules would not exceed serum concentrations of 1 % and typically would be less than 0.1 %. Serum levels are expected to drop over time due to enzymatic breakdown of the components. Independent in vivo pharmacokinetic studies indicate that the active components in ASEA have approximately a 17 minute half-life in the blood and thus would be effectively cleared from the blood within a few hours. Thus no toxic response is expected due to exposure of healthy human cells at such levels. It has been seen in these in vitro studies that direct exposure of human cells to serum concentrations of up to 20% is still well tolerated. The complete lack of toxicity, comparable to the PBS control, is extremely rare and indicates that despite the reactivity of this mixture, it is well tolerated by human tissues and is native to or compatible with the extracellular environments.
- the raw data reflects more than a 10 fold increase in antioxidant activity related to ASEA infusion. Taking into account experimental uncertainties, it is 98% certain that the serum infusion of small concentrations ( ⁇ 1 %) of a composition of the invention increased antioxidant efficiencies by at least 800%. Further investigations should be done to confirm this increase and explore concentration dependence for these low-level serum concentrations.
- Results of First-Attempt Methods to Determine SOD activity for high serum a composition of the invention concentration Diluted lysates showed a marginal increase in enzymatic activity associated with treatment with a composition of the invention. Changes in enzymatic activity were marginal in the initial range of 5-10% a composition of the invention (final concentration, v/v).
- the data represent the first attempt to measure SOD activity using primary HMVEC-L cells treated with a composition of the invention. It is feasible that the lack of SOD activity associated with 5-10% a composition of the invention might be related to non-specific inhibition at high dose.
- the primary concern is that we have little understanding of the primary human HMVEC-L cell model and cannot determine whether these cells are optimal for investigating antioxidant defense regulation induced by a composition of the invention.
- ascorbic acid known to break down certain redox signaling complexes in A a composition of the invention
- some modification of the medium formula such as omission of ascorbic acid for short periods of time defined empirically
- Initial efforts to serum-starve these cells were unsuccessful and resulted in extensive cell death over 24 hours, indicating that the cells are dependent on the growth factors supplemented in the cell culture medium to maintain cell viability.
- HMVEC-L cells were again thawed and maintained according to manufacturer's directions.
- the culture medium contained epidermal growth factor, hydrocortisone, GA-1000, fetal bovine serum, vasoactive endothelial growth factor, basic fibroblast growth factor, insulin growth factor-1 and ascorbic acid in randomly cycling cultures. Ascorbic acid was withheld from serum-starved cultures.
- HMVEC-L Cell cultures in both normal random cell cycles and in serum starvation were exposed to high-concentration (5-20%) and low-concentration (1 %) ASEA in the serum medium and analyzed in conjunction with cultures exposed only to phosphate buffered saline solution (PBS), as a negative control.
- PBS phosphate buffered saline solution
- Computer automated imaging techniques were used to determine the relative degree of nuclear accumulation of NRF2 via fluorescent analysis over several cells.
- NRF2 regulates the transcription of a number of phase II antioxidant defense enzymes and raises the possibility that additional antioxidant defense enzymes, such as glutathione transferase, may be expressed through exposure to ASEA.
- additional antioxidant defense enzymes such as glutathione transferase
- Results of HMVEC-L Nuclear Accumulation of NRF2 Initial screen of human endothelial cells suggests a subpopulation of cells showed increased nuclear staining pattern (focal) following treatment with high-concentrationof a composition of the invention. The positions of nuclei are indicated by DAPI stain in lower panel. Foci appear brighter in a composition of the invention stimulated cells which indicates higher level of NRF2 transcription factor in the nucleus. H 2 0 2 was used as positive control. This effect was difficult to quantify based on nuclear staining pattern. ( Figure 29).
- Typical cell images are shown below for indicated cell cultures exposed to low- concentrations of a composition of the invention. Accumulation of NRF2 into the nucleus was clearly seen in serum-starved cell cultures exposed to low-concentrations of a composition of the invention. Automated analysis revealed strong time- dependent nuclear accumulation of NRF2 in serum-starved cells, relative to the negative control, at the 30 and 60 minute time points (Figure 30).
- the nuclear staining profile was qualitatively different from the cells maintained in optimal growth medium (randomly cycling group). There was weak qualitative nuclear accumulation of NRF2 induced by exposure to a composition of the invention in these cells at 30, 60 and 120 minute time points, and yet the effect was not nearly as pronounced as in the serum-starved cultures. However, serum-starvation induced significant cell death complicating interpretation of the data. The trends appeared weak and require validation by Western Blot.
- NRF2 levels were increased in a time-dependent fashion in nuclear extracts prepared from HMVEC-L cells treated with 1 % ASEA. H 2 0 2 (30 min) did not increase nuclear NRF2 levels.
- HMVEC-L cells were treated with 5-20% ASEA for 72 hr and cell number was determined using a Coulter Counter. Control (0 concentration group) was treated with 20% PBS. Serum LDH levels were also measured as an indicator of cell culture viability at 0 to 20% concentration of the compositions of the invention / serum concentrations. Recall that lower serum LDH concentrations indicate less cell membrane failure. Similar experiments were performed for murine (JB6) epidermal cells.
- Serum LDH levels were obtained as an indication of membrane integrity and Neutral Red dye was used as an indication of lysosomal integrity. Recall that as cell membranes fail, LDH is released into the serum medium. Lower quantities of LDH indicate higher cell viability. The integrity of lysosomes, necessary for viable cell function, are measured by absorption of Neutral Red dye stain. Higher quantities of Neutral Red absorbance indicate higher cell viability.
- Results of HMVEC-L viability exposed high-concentration composition of the invention and to escalating amounts of Cachexin stressor (Figure 34): Both confluent (A2) and normal (pS) HMVEC-L cultures exhibited up to 30% improvement (relative to PBS controls) in LDH levels related to exposure to compositions of the invention after acute (up to 5 nm/mL) Cachexin insult.
- the LDH data suggest that HMVEC-L cells stressed by Cachexin are less likely to die due to cell membrane failure after being exposed to compositions of the invention.
- HMVEC-L cell cultures prepared in two phases, in the confluent end-of-life-cycle A2 phase (a phase typically insensitive to Cachexin insult) and in the normal random cycle pS phase were exposed for 24 hours to serum concentrations (v/v of 2.5%, 5%, 10%, 15% and 20%) of either the PBS control or a composition of the invention. Cachexin responsiveness was then determined by monitoring LDH activity in both the intracellular cytosol and in the surrounding growth media.
- LDH release cell membrane rupture and death
- intracellular LDH activity indicates loss of cellular integrity.
- composition of the invention exposure did not affect cell death.
- composition of the invention exposure clearly amplified the Cachexin reception rapidly decreasing cellular function and there were also clear indications of concentration-dependent cell death. There is strong evidence that exposure to compositions of the invention increases Cachexin responsiveness in the A2 cell cultures.
- compositions of the invention significantly increases Cachexin responsiveness in A2 and borderline pS HMVEC-L cell cultures.
- exposure to compositions of the invention alone might decrease integrity of cellular LDH activity in A2 type cells; recall that zero toxic response was detected in randomly cycling cells even under large concentrations, so effects due to toxicity are not expected in normal cells.
- exposure to compositiosn of the invention may tend to accelerate the removal of non-responsive confluent cells. This is evidently true when Cachexin is present.
- These results might also bear on the observations that exposure to compositions of the invention seemed to diminish cell proliferation in high concentrations. No such trend was tried for low-concentration exposure. Note that it is difficult to discount the possibility that high-concentration effects might simply be artifacts due to the interference of compositions of the invention with the growth medium.
- Murine (JB6) cell cultures were subjected to high-level radiation exposure (X-rays) and, in a separate investigation, cultures were subject to serum starvation of growth factors for 24 hours. The cells were then exposed to 5-10% ASEA exposure as means to determine the effect of composition of the invention exposure on such stressed cells. Cell counts were taken before and after composition of the invention exposure.
- results of effects of 5-10% composition of the invention exposure on radiation and serum-starved murine cells Quantitative analysis was not compiled for these experiments. Qualitative analysis, however, reveals results that might be of some interest. For the radiation-damaged culture, immediate cell death was observed for more than half of the culture upon exposure to composition of the invention. No further cell-death was seen thereafter. Upon inspection under a microscope, the remaining living cells appeared normal and healthy. It appears that exposure to a composition of the invention may have helped accelerate cell death among the more seriously damaged cells and allowed for the survival of healthy or repairable cells.
- compositions of the invention at a lower concentration induces a 20-30% increase in the nuclear translocation of the NRF2 transcription factor in HMVEC-L cells that appears to be transient (30-60 min).
- a composition of the invention induced a parallel decrease in the phosphorylation of an extra- nuclear protein whose phosphorylation status is clearly increased in response to hydrogen peroxide treatment, consistent with an antioxidant mode of action.
- HMVEC-L cells When stressed with Cachexin depends upon cell phase. Normal randomly cycling HMVEC-L cells (pS) exhibited typical behavior when stressed with Cachexin: exhibiting decrease in cell viability accompanied by cell death. Confluent end-of-life-cycle (A2) and borderline HMVEC-L cells, as expected, were less sensitive to Cachexin insult, exhibiting less pronounced decreases in cell viability and less cell death.
- compositions of the invention caused no significant change in the response of the normal random cycling pS cells to Cachexin (showing similar loss of cell viability and cell-death).
- A2 cell cultures exposed to a composition of the invention exhibited increased sensitivity to Cachexin, restoring behavior similar to that of normal cells. This behavior was reinforced as concentration dependence was examined. Borderline A2 cells, exhibiting a relatively small Cachexin response, and A2 cells that are normally insensitive to Cachexin insult, exhibited a much stronger response to Cachexin when exposed to compositions of the invention, both in decrease in viability and increased cell death.
- compositions of the invention causes increased rates of A2 cell death, enhancing the natural reception of Cachexin in such end-of-life-cycle cells. Yet exposure to composition of the invention is not expected to cause any change in normal cell viability.
- Cachexin is normally secreted to instigate cell death in damaged or dysfunctional tissues, allowing surrounding healthy cells to divide and fill in voids. Thus, increasing the sensitivity to Cachexin in dysfunctional cells may help accelerate such a process and is not always deleterious.
- compositions of the invention The infusion of a certain balanced mixture of redox signaling molecules using compositions of the invention into viable HMVEC-L and JB6 cell cultures has been seen to elicit distinct bioactivity. No indications of toxicity or the expression of inflammatory cytokines were observed and yet there was increased antioxidant and protective enzyme expression (as evidenced by increased nuclear NRF2) and greatly increased efficacy for the two master antioxidants, GPx and SOD. This behavior suggests that infusion with compositions of the invention might tend to induce and enhance oxidative defense mechanisms without inducing toxic or inflammatory responses in such cells. Such action is unprecedented or extremely rare. Normally, low-level toxicity induces slight oxidative stress and inflammatory response which in turn induces oxidative defense and cell repair mechanisms. It would be of interest to determine concentration dependency of this effect with ultra-low-concentration infusions of compositions of the invention.
- compositions of the invention should also be explored. Natural healing processes involve a repair or replace mechanism by which marginally damaged cells are repaired, when possible, or undergo apoptosis, programmed death, if they cannot be repaired and then are replaced through mitosis of healthy neighboring cells. It is fairly evident that infusion of composition of the invention, of itself, is not causing direct stress to exposed cells, however, it might tend to increase the efficiency of certain cytokine "death domain" messengers (Cachexin) that are designed to induce cell death in dysfunctional or damaged cells.
- Cachexin cytokine "death domain" messengers
- the nuclear translocation of NRF2 can be considered part of the phase II oxidative defense response which includes expression of antioxidants, DNA repair molecules and other known repair mechanisms.
- Apoptosis is part of the replace mechanism when cells have undergone unrepairable damage and must be removed and replaced. Both antioxidant defense and apoptotic mechanisms are central to normal tissue repair and regeneration. Redox signaling is involved in several of the pathways, such as p53 gene expression, that can determine whether a cell undergoes apoptosis or not. Chronic oxidative stress tends to favor cell death. Certainly the presence of Cachexin and other death domain messengers favor cell death. The observation that infusion with compositions of the invention enhances Cachexin reception might indicate that infusion with compositions of the invention also might serve to enhance reception of messengers in the signaling process that determines whether defense, repair or replace mechanisms are activated.
- a 42 year old man diagnosed with type 2 diabetes is treated by intravenously injecting a combination product comprising a PPAR inhibitor (an eicosanoid) and a redox signaling agent (RXN) composition of the invention. After about 1 week, the patient maintains a more regular glucose profile with minimal glucose excursions as compared to prior to treatment.
- PPAR inhibitor an eicosanoid
- RXN redox signaling agent
- a 22 year old woman diagnosed with type 1 diabetes is treated by intravenously injecting a combination product comprising a PPAR inhibitor (a thazolidinedione) and a redox signaling agent (RXN) composition of the invention. After about 1 week, the patient maintains a more regular glucose profile with minimal glucose excursions as compared to prior to treatment.
- a PPAR inhibitor a thazolidinedione
- RXN redox signaling agent
- a 72 year old man diagnosed with type 2 diabetes is treated by orally administering a combination product comprising a PPAR inhibitor (an eicosanoid) and a redox signaling agent (RXN) composition of the invention. After about 1 week, the patient maintains a more regular glucose profile with minimal glucose excursions as compared to prior to treatment.
- PPAR inhibitor an eicosanoid
- RXN redox signaling agent
- a 31 year old woman diagnosed with oral cancer is treated by orally administering a combination product comprising a PPAR inhibitor (a fibrate) and a redox signaling agent (RXN) composition of the invention. After about 1 month, the patient shows a regression in her symptoms.
- a PPAR inhibitor a fibrate
- RXN redox signaling agent
Abstract
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---|---|---|---|---|
CN104846075A (en) * | 2015-03-31 | 2015-08-19 | 温州医科大学 | Method for evaluating in-vivo oxidation stress based on detection of peripheral blood mitochondria DNA oxidative damage |
US9332742B2 (en) | 2012-03-16 | 2016-05-10 | Regeneron Pharmaceuticals, Inc. | Histidine engineered light chain antibodies and genetically modified non-human animals for generating the same |
US9648856B2 (en) | 2012-03-16 | 2017-05-16 | Regeneron Pharmaceuticals, Inc. | Non-human animals expressing pH-sensitive immunoglobulin sequences |
Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9486481B2 (en) | 2007-10-30 | 2016-11-08 | Reoxcyn Discoveries Group, Inc. | Method of modulating fatty acid mobilization and oxidation |
US9309601B2 (en) * | 2012-10-16 | 2016-04-12 | GenEon Technologies LLC | Electrochemical activation of water |
US20150099010A1 (en) | 2013-10-07 | 2015-04-09 | Reoxcyn Discoveries Group, Inc | Redox signaling gel formulation |
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CA3058806A1 (en) * | 2017-04-03 | 2018-10-11 | Coherus Biosciences Inc. | Ppar.gamma. agonist for treatment of progressive supranuclear palsy |
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20040224995A1 (en) * | 2003-05-09 | 2004-11-11 | University Of North Texas Health Science Center At Fort Worth | Neuroprotective effects of PPARy agonists against cellular oxidative insults |
WO2011066659A1 (en) * | 2009-12-04 | 2011-06-09 | Technologie Biolactis Inc. 15468 | Method of regulating ppar, obesity related pathways and their associated metabolic impact |
US20120070511A1 (en) * | 2009-03-23 | 2012-03-22 | University Of Miami | Mitochondrial inhibitors and uses thereof |
Family Cites Families (37)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3984303A (en) * | 1975-07-02 | 1976-10-05 | Diamond Shamrock Corporation | Membrane electrolytic cell with concentric electrodes |
US4177116A (en) | 1977-06-30 | 1979-12-04 | Oronzio DeNora Implanti Elettrochimici S.p.A. | Electrolytic cell with membrane and method of operation |
IT1114820B (en) * | 1977-06-30 | 1986-01-27 | Oronzio De Nora Impianti | ELECTROLYTIC MONOPOLAR MEMBRANE CELL |
US5234563A (en) * | 1992-06-01 | 1993-08-10 | Janix Kabushiki Kaisha | Electrolytic ionized water producer of a continuous type |
JP3285978B2 (en) * | 1992-11-27 | 2002-05-27 | ティーディーケイ株式会社 | Electrode for drinking water electrolysis, method for producing the same, and ion water generator |
JP2737643B2 (en) * | 1994-03-25 | 1998-04-08 | 日本電気株式会社 | Method and apparatus for producing electrolytically activated water |
JP3468835B2 (en) * | 1994-05-09 | 2003-11-17 | ホシザキ電機株式会社 | Electrolyzed water generator |
US6117285A (en) | 1994-08-26 | 2000-09-12 | Medical Discoveries, Inc. | System for carrying out sterilization of equipment |
EP1284728A4 (en) * | 1999-10-22 | 2004-05-19 | Merck & Co Inc | Pharmaceuticals for treating obesity |
US6572902B2 (en) * | 2001-04-25 | 2003-06-03 | Advanced H2O, Inc. | Process for producing improved alkaline drinking water and the product produced thereby |
GB0119480D0 (en) * | 2001-08-09 | 2001-10-03 | Jagotec Ag | Novel compositions |
EP1579037A4 (en) * | 2002-03-06 | 2008-02-13 | Univ Georgia Res Found | Method and apparatus for electrolyzing water |
WO2004007376A1 (en) * | 2002-07-17 | 2004-01-22 | Peter Nunn | Water treatment device for electrolyzing, magnetizing, and re-resonating water |
JP2004058006A (en) * | 2002-07-31 | 2004-02-26 | First Ocean Kk | Method of manufacturing electrolytic water |
US7230016B2 (en) * | 2003-05-13 | 2007-06-12 | Synthon Ip Inc. | Pioglitazone salts, such as pioglitazone sulfate, and pharmaceutical compositions and processes using the same |
JP2008501027A (en) * | 2004-05-28 | 2008-01-17 | メルク エンド カムパニー インコーポレーテッド | Benzourea with anti-diabetic activity |
US20060040876A1 (en) * | 2004-06-10 | 2006-02-23 | Rong-Hwa Lin | Modulation of peroxisome proliferator-activated receptors |
RU2252920C1 (en) * | 2004-06-28 | 2005-05-27 | Государственное научное учреждение Поволжский научно-исследовательский институт эколого-мелиоративных технологий | Plant for electrochemical activation of drinking and sprinkling water |
WO2006042082A2 (en) * | 2004-10-08 | 2006-04-20 | Electric Aquagenics Unlimited | Apparatus and method for producing electrolyzed water |
US20080096915A1 (en) * | 2005-01-13 | 2008-04-24 | Greenberg Traurig LLP | Compositions for the treatment of metabolic disorders |
KR101532778B1 (en) * | 2005-03-23 | 2015-06-30 | 오클루스 이노바티브 사이언시즈 인코포레이티드 | Method of treating second and third degree burns using oxidative reductive potential water solution |
WO2006107760A1 (en) * | 2005-04-01 | 2006-10-12 | Electric Aquagenics Unlimited | Electrolyzed water treatment for poultry products |
RU2008122547A (en) * | 2005-11-07 | 2009-12-20 | Айрм Ллк (Bm) | COMPOUNDS AND COMPOSITIONS AS MODULATORS OF ARPP (ACTIVATED RECEPTORS OF PROLIFERATOR PEROXIS) |
US8025787B2 (en) * | 2006-02-10 | 2011-09-27 | Tennant Company | Method and apparatus for generating, applying and neutralizing an electrochemically activated liquid |
DE102006028168A1 (en) * | 2006-06-16 | 2007-12-20 | Uhde Gmbh | Apparatus for electrochemical water treatment |
PE20090159A1 (en) * | 2007-03-08 | 2009-02-21 | Plexxikon Inc | INDOL-PROPIONIC ACID DERIVED COMPOUNDS AS PPARs MODULATORS |
JP5563824B2 (en) * | 2007-10-24 | 2014-07-30 | サントリーホールディングス株式会社 | Peroxisome proliferator-responsive receptor (PPAR) ligand agent |
US8367120B1 (en) * | 2007-10-31 | 2013-02-05 | Reoxcyn Discoveries Group, Inc. | Method and apparatus for producing a stablized antimicrobial non-toxic electrolyzed saline solution exhibiting potential as a therapeutic |
US20090110749A1 (en) * | 2007-10-30 | 2009-04-30 | Medical Management Research, Inc. | Method and apparatus for producing a stabilized antimicrobial non-toxic electrolyzed saline solution exhibiting potential as a therapeutic |
JP2011525146A (en) * | 2008-06-19 | 2011-09-15 | テナント カンパニー | Electrolytic scale removal method with constant output |
CN101371807A (en) * | 2008-07-19 | 2009-02-25 | 冯文昌 | Apparatus for curing cancer with water temperature and oxygen |
US20110100889A1 (en) * | 2008-07-31 | 2011-05-05 | Mitsubishi Electric Corporation | Sterilzation and anti-bacterialzation equipment |
FR2940289B1 (en) * | 2008-12-23 | 2014-09-12 | Biopharmed | AMINO HYDROXYQUINOLINE CLASS DERIVATIVES FOR THE TREATMENT OF PANCREATIC CANCER |
WO2010151816A1 (en) * | 2009-06-26 | 2010-12-29 | Eric Kuhrts | Water-soluble dietary fatty acids |
WO2011120923A1 (en) * | 2010-03-30 | 2011-10-06 | Boehringer Ingelheim International Gmbh | Pharmaceutical composition comprising an sglt2 inhibitor and a ppar- gamma agonist and uses thereof |
JP5770449B2 (en) * | 2010-10-20 | 2015-08-26 | 株式会社Kri | PPARα activator |
CN102320684B (en) * | 2011-08-25 | 2013-05-29 | 洪韫麒 | Reactor for continuously generating water with high oxidation potential and high reduction potential |
-
2012
- 2012-12-07 MX MX2015003700A patent/MX2015003700A/en unknown
- 2012-12-07 RU RU2015114800A patent/RU2636483C2/en not_active IP Right Cessation
- 2012-12-07 CN CN201280076550.0A patent/CN104903494B/en not_active Expired - Fee Related
- 2012-12-07 JP JP2015533035A patent/JP6314142B2/en active Active
- 2012-12-07 WO PCT/US2012/068613 patent/WO2014046697A1/en active Application Filing
- 2012-12-07 EP EP12884838.9A patent/EP2898116A4/en not_active Withdrawn
- 2012-12-07 US US14/430,486 patent/US9962404B2/en not_active Expired - Fee Related
-
2013
- 2013-02-28 EP EP16190337.2A patent/EP3130564A1/en not_active Withdrawn
- 2013-02-28 RU RU2015114716A patent/RU2015114716A/en not_active Application Discontinuation
- 2013-02-28 EP EP13839705.4A patent/EP2897613A4/en not_active Withdrawn
- 2013-02-28 US US14/430,506 patent/US20150246071A1/en not_active Abandoned
- 2013-02-28 JP JP2015533038A patent/JP2015530405A/en active Pending
- 2013-02-28 WO PCT/US2013/028420 patent/WO2014046722A1/en active Application Filing
- 2013-02-28 MX MX2015003701A patent/MX2015003701A/en unknown
- 2013-02-28 CN CN201380054979.4A patent/CN104936594A/en active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20040224995A1 (en) * | 2003-05-09 | 2004-11-11 | University Of North Texas Health Science Center At Fort Worth | Neuroprotective effects of PPARy agonists against cellular oxidative insults |
US20120070511A1 (en) * | 2009-03-23 | 2012-03-22 | University Of Miami | Mitochondrial inhibitors and uses thereof |
WO2011066659A1 (en) * | 2009-12-04 | 2011-06-09 | Technologie Biolactis Inc. 15468 | Method of regulating ppar, obesity related pathways and their associated metabolic impact |
Non-Patent Citations (3)
Title |
---|
LAVROVSKY, Y. ET AL.: "Role of redox-regulated transcription factors in inflammation, aging and age-related diseases", EXPERIMENTAL GERONTOLOGY, vol. 35, 2000, pages 521 - 532, XP055235701 * |
MICHALIK, L. ET AL.: "PPARs mediate lipid signaling in inflammation and cancer", PPAR RESEARCH, vol. 2008, 2008, XP055235706 * |
See also references of EP2897613A4 * |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9332742B2 (en) | 2012-03-16 | 2016-05-10 | Regeneron Pharmaceuticals, Inc. | Histidine engineered light chain antibodies and genetically modified non-human animals for generating the same |
US9648856B2 (en) | 2012-03-16 | 2017-05-16 | Regeneron Pharmaceuticals, Inc. | Non-human animals expressing pH-sensitive immunoglobulin sequences |
US9801362B2 (en) | 2012-03-16 | 2017-10-31 | Regeneron Pharmaceuticals, Inc. | Non-human animals expressing pH-sensitive immunoglobulin sequences |
CN104846075A (en) * | 2015-03-31 | 2015-08-19 | 温州医科大学 | Method for evaluating in-vivo oxidation stress based on detection of peripheral blood mitochondria DNA oxidative damage |
Also Published As
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CN104903494A (en) | 2015-09-09 |
RU2015114800A (en) | 2016-11-10 |
US20150246832A1 (en) | 2015-09-03 |
CN104936594A (en) | 2015-09-23 |
EP2898116A1 (en) | 2015-07-29 |
EP2897613A1 (en) | 2015-07-29 |
WO2014046697A1 (en) | 2014-03-27 |
JP6314142B2 (en) | 2018-04-18 |
EP3130564A1 (en) | 2017-02-15 |
JP2015534608A (en) | 2015-12-03 |
JP2015530405A (en) | 2015-10-15 |
CN104903494B (en) | 2017-11-03 |
EP2897613A4 (en) | 2016-03-30 |
US9962404B2 (en) | 2018-05-08 |
MX2015003701A (en) | 2015-10-29 |
RU2015114716A (en) | 2016-11-10 |
MX2015003700A (en) | 2015-10-30 |
RU2636483C2 (en) | 2017-11-23 |
EP2898116A4 (en) | 2016-06-01 |
US20150246071A1 (en) | 2015-09-03 |
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