Classifications

• • 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
• • 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
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61J—CONTAINERS SPECIALLY ADAPTED FOR MEDICAL OR PHARMACEUTICAL PURPOSES; DEVICES OR METHODS SPECIALLY ADAPTED FOR BRINGING PHARMACEUTICAL PRODUCTS INTO PARTICULAR PHYSICAL OR ADMINISTERING FORMS; DEVICES FOR ADMINISTERING FOOD OR MEDICINES ORALLY; BABY COMFORTERS; DEVICES FOR RECEIVING SPITTLE
  • A61J1/00—Containers specially adapted for medical or pharmaceutical purposes
  • A61J1/05—Containers specially adapted for medical or pharmaceutical purposes for collecting, storing or administering blood, plasma or medical fluids ; Infusion or perfusion containers
  • A61J1/10—Bag-type containers
• • 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
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
  • A61K35/02—Medicinal preparations containing materials or reaction products thereof with undetermined constitution from inanimate materials
  • A61K35/08—Mineral waters; Sea water
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
  • A61K8/19—Cosmetics or similar toiletry preparations characterised by the composition containing inorganic ingredients
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K8/00—Cosmetics or similar toiletry preparations
  • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
  • A61K8/19—Cosmetics or similar toiletry preparations characterised by the composition containing inorganic ingredients
  • A61K8/20—Halogens; Compounds thereof
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
  • A61Q19/00—Preparations for care of the skin
  • A61Q19/08—Anti-ageing preparations
• • 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
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
  • C12N5/0018—Culture media for cell or tissue culture
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
  • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
  • A61K2800/83—Electrophoresis; Electrodes; Electrolytic phenomena
• • 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
  • C02F2001/46133—Electrodes characterised by the material
  • C02F2001/46138—Electrodes comprising a substrate and a coating
  • C02F2001/46142—Catalytic coating
• • 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
  • C02F2001/46133—Electrodes characterised by the material
  • C02F2001/46138—Electrodes comprising a substrate and a coating
  • C02F2001/46147—Diamond coating
• • C—CHEMISTRY; METALLURGY
  • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
  • C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
  • C02F2103/02—Non-contaminated water, e.g. for industrial water supply
  • C02F2103/026—Treating water for medical or cosmetic purposes
• • 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
  • C02F2201/4612—Controlling or monitoring
  • C02F2201/46125—Electrical variables
• • 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
  • C02F2201/4612—Controlling or monitoring
  • C02F2201/46125—Electrical variables
  • C02F2201/4614—Current
• • 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
  • C02F2201/4612—Controlling or monitoring
  • C02F2201/4615—Time

Definitions

• the subject of the invention is the field of the treatment of diseases or disorders linked to cellular senescence, in particular the treatment of cancer or of neurodegenerative diseases.
• the invention relates more particularly to a process for obtaining an electrolyzed water useful for the treatment of diseases or disorders linked to cell senescence.
• the invention also relates to the water obtained by this process, its use for the treatment of diseases or disorders linked to cellular senescence by direct application or in the form of compositions, and a suitable device for treating said conditions.
• the invention also relates to a method of selecting non-senescent cells useful in transplant applications
• Cellular senescence is generally understood as a definitive arrest of the cell cycle resulting from various causes.
• the cell cycle arrest mechanism induced by cell senescence is complex and involves interactions between telomere shortening, cell inflammation and stress.
• Application WO2018215795 will also be cited, which relates to an agent intended to be used for selectively killing one or more senescent cells, said agent being chosen from the following elements: a cardiac glycoside or aglycone, a focal adhesion kinase inhibitor (F AK), an HMG-CoA reductase inhibitor, JF D00244, cyclosporine, tyrphostine AG879, cantharidin, diphenyleneiodonium chloride, rottlerin, 2,3-dimethoxy-1, 4-naphthoquinone, LY-367,265, rotenone, idarubicin, dequalintum chloride, vincristine, nitazoxanide, nitrofurazone, temsirolimus, eltrombopag, adapalene, azacyclonol, enoxacin and raltegravir, and their salts pharmaceutically acceptable.
• F AK
• Application US2018256568 will also be cited which relates to a method of treating a disease or a disorder in a target tissue in a subject, in which the symptoms of the disease or of the disorder are caused at least in part. by senescent cells located in or around the target tissue, wherein the target tissue is cancer free, and wherein the senescent cells are defined as cells expressing the p16 protein.
• the method comprises the administration in or around the target tissue of a series of treatments with a pharmaceutical composition which comprises a senolitic agent which contacts the senescent cells located in or around the target tissue and selectively removes these cells. senescent without affecting, if possible, adjacent non-senescent cells.
• a pharmaceutical composition which comprises a senolitic agent which contacts the senescent cells located in or around the target tissue and selectively removes these cells. senescent without affecting, if possible, adjacent non-senescent cells.
• the application describes a whole series of chemical molecules making it possible to treat senescent cells.
• one of the objects of the invention is the envisaged use of electrolyzed water according to the invention in the treatment of diseases linked to cell senescence, such as in particular cancers or neurodegenerative diseases.
• the electrolyzed water according to the invention is obtained by the implementation of a three-step process which begins with the use of running or spring water optionally added with a sulphate salt, carbonates or any other compatible salt known to those skilled in the art at a concentration of 0.5 to 2 g / L followed by electrolysis of said water by an electrolysis module comprising at least one boron-doped diamond electrode fixed on a silicon substrate in wherein the boron concentration is between 200 (B 3x10 19 atoms / cm 3) and 2000 ppm (B 3,52x1 ° atoms / cm 3 B), in particular between 200 ppm (B 3x1019 atoms / cm3) and 1500 ppm (2x1020 B atoms / cm3), the said module subjecting the water to a quantity of current during the electrolysis process of less than 500 mAh / L of water, more preferably between 40 and 250 mAh / L of water, even more preferably between 50 and 200 mAh / L, the duration of the electrolysis
• the electrolysis can be done all at once, continuously, or even cyclically with a number of cycles between 2 and 12 cycles over a period of 24 hours, two consecutive cycles being spaced at least 30 minutes.
• a subject of the invention is also electrolyzed water obtained by the method described above.
• a subject of the invention is also a method for increasing the senolitic character of senescent biological cells present in a cellular medium, characterized in that it consists in introducing into the cell medium an electrolyzed water obtained according to the method described above. .
• This process consists in particular in increasing at least the beta-galactosidase (SA- -Gal) activity associated with senescence by contacting of said cells with the electrolyzed water thus obtained.
• SA- -Gal beta-galactosidase
• the inventors have in fact discovered that the electrolyzed water obtained according to the invention had the singular property of significantly increasing this beta-galactosidase (SA- -Gal) activity when senescent cells such as present in particular in fibroblasts were introduced. in contact with said water.
• the percentage of water contained in the cell medium is strictly greater than 0, and preferably greater than 5%, more preferably still greater than 25%.
• the senescent cells are maintained in the cell medium in contact with water for a period of more than 1 min.
• Such an increase in the senolitic character of senescent cells has the major advantage of allowing an induced selection of healthy cells relative to senescent cells by acceleration of the natural process of senescence in a cellular medium or biological tissue, and therefore the accelerated disappearance. said cells for the benefit of healthy cells. This opens up exceptional possibilities for using the water of the invention for prophylactic purposes, to aid in therapeutic treatment, or even for direct therapeutic treatment.
• a subject of the invention is also a composition comprising an electrolyzed water obtained according to the method described above for use as a medicament in the treatment of disorders linked to senescence.
• the disorders targeted by the inventive use include diseases linked to senescence and in particular certain cancers or neurodegenerative diseases.
• composition comprising electrolyzed water according to the invention consists of 95 to 100% electrolyzed water and 0 to 5% of an excipient and / or an emulsifier.
• composition according to the invention can be in the form of a cream, a gel, a dressing or else a patch. Furthermore, when the composition according to the invention can be exclusively composed of electrolyzed water, the latter can be added to a device containing it such as a reservoir or an infusion bag containing pure electrolyzed water or in the form of liquid composition.
• Electrolyzed water is obtained by means of an electrolysis module allowing the implementation of the method described above, said module being able to be present in a fixed or mobile manner in a device such as a tank, said tank being able to be used to fill infusion bags on a production line for example.
• a conventional water distribution circuit preferably at the distribution outlet like a tap, if one considers wanting to prepare a drink from electrolyzed water, the latter then being able to be consumed for the purpose of treating disorders linked to cellular senescence by the oral route.
• a subject of the invention is also a method for selecting non-senescent cells in a cell extract or a biological tissue comprising several cells by bringing them into contact with an electrolyzed water obtained by carrying out the method according to invention for a period of 1 to 30 minutes.
• obtaining a cell extract or a healthy tissue free in whole or in large part from senescent cells may find its interest in the use for the treatment of grafts, in particular of skin or liver.
• FIG.1 shows the efficiency of electrolyzed water according to the invention named WDW on chondrosarcoma-type tumor cell lines in comparison with unelectrolyzed ultra-pure MQW water.
• FIG.2 shows the superiority of the electrolyzed water obtained according to the process of the invention, on the modification of the SA- -Gal activity, the increase of which is witness to the presence of senescent cells.
• senescence is a complex process which causes a slow degradation of the functions of the cell at the origin of the aging of organisms. This is a complex biological mechanism, involving the degradation of many genes, especially in senescent cells. Usually the telomeric part of the chromosomes is affected.
• telomere is a highly repetitive region of DNA present at the end of a chromosome. Each time a rod chromosome of a eukaryote is replicated, during mitosis also called cell division, the enzymatic complex of DNA polymerase is unable to copy the last few nucleotides. Telomeres shorten with age, likely as a result of the repeated inflammation and / or stress that organisms undergo. This results in the appearance within an organism, in which senescent cells are present in large numbers, diseases such as age-related cancer or neurodegenerative diseases.
• senescent cells stop dividing unlike normal cells in the body, but the fact that these cells no longer divide also has the consequence that the mode of functioning of these cells differs from healthy or normal cells.
• the loss of competent cells in terms of proliferation can be responsible for a pathology, as is suspected in glaucoma, cataracts, pancreatic diabetes or even osteoarthritis.
• the inflammation caused by SASP can play a causal role in the onset of diseases; senescent cells with significant inflammatory potential gradually move into the tissue environment where they will somehow 'contaminate' healthy cells nearby which in turn will become senescent as suspected in diseases such as atherosclerosis, cardiovascular disease or even age-related cancers.
• the present invention relating to a particular electrolyzed water, obtained by a process involving particular electrodes of boron doped diamond fixed to silicon, makes it possible to solve, at least partially, the problems or drawbacks mentioned above.
• the electrolysis module for implementing the method for preparing electrolyzed water according to the invention comprises at least one, preferably at least two boron-doped diamond electrodes which are fixed on a silicon substrate.
• each electrode is advantageously between and 10 and 100 cm 2 , preferably 60 and 80, more preferably around 70 cm 2 .
• the boron doping of the diamond electrode also has its importance on the properties of the water obtained; the boron concentration is between 200 ppm (3x10 19 B atoms / cm 3 ) and 1500 ppm (2x10 20 B atoms / cm 3 ).
• This boron concentration as well as the nature of the diamond-based electrodes on silicon gives it properties allowing it to operate on a potential between -1 V and -2 V on the cathodic polarization and on + 2V and + 4V on anodic polarization in comparison to a hydrogen reference electrode.
• the electrolysis module is connected to a power supply module and is open to a flow of water which will pass through it.
• the electrodes are supplied with direct current by the power supply module which is connected to the electrolysis module providing direct current to the electrode (s).
• the supply current is set between 1.5A and 7A. If reverse polarity is required, this can be done automatically by the power module.
• the water can come from different sources but must pass through the electrolysis module because it cannot operate in the absence of water.
• the electrolysis module can be permanently crossed by water depending on the device in which it is located.
• An internal measurement system comprising a hydraulic flow sensor is present, interacting with the power supply module and which allows the standby, stopping or starting of the electrolysis module in the absence or presence of water, and therefore activate electrolysis.
• the water electrolysis module according to the present invention can advantageously operate in automatic mode or can be activated or deactivated on demand, manually or using a remote control system.
• the water electrolysis module does not necessarily need permanent activation, but can usefully be activated cyclically, that is to say. say at convenient intervals, preferably but not necessarily at regular intervals. It has been found that electrolysis of water at regular intervals allows the water to remain therapeutically active for a long time. Electrolysis cycles can in particular be carried out at a rate of 2 to 12 cycles over a period of 24 hours, two consecutive cycles being spaced at least 30 minutes apart, which makes it possible to obtain water with a therapeutic potential.
• the process for preparing electrolyzed water according to the invention using the module described above comprises the following steps: a.
• a conductivity salt is added at a concentration of 0.5 to 2 g / L b.
• the amount of current delivered during the electrolysis process is between 15 and 500 mAh / L of water, more preferably 40 to 250 mAh / L of water, even more preferably 50 to 200 mAh / L ; the electrolysis time being between 1 and 60 minutes.
• the method of the invention thus makes it possible to obtain clean water for use in the treatment of diseases or disorders linked to cell senescence.
• the treatment of diseases or disorders linked to cellular senescence can, without this being limiting, include the treatment of diseases linked to age such as neurodegenerative diseases, skin conditions but also glaucoma, cataracts, pancreatic diabetes, osteoarthritis, atherosclerosis, cardiovascular disease or even age-related cancers.
• the water obtained according to the method of the invention can also be useful in the case of grafting or transplantation of tissue such as the liver or even an organ or even skin grafting.
• the water thus obtained can also be used in different forms or compositions or within different devices.
• the devices used can be selected from a reservoir or else an infusion bag comprising electrolyzed water as described above.
• the electrolysis module used will then be integrated into an existing water circuit in order to prepare the electrolyzed water; one can very well also imagine the electrolysis module connected to or immersed in a tank. Any water, whether natural, from a source or from a municipal network, can be used in the process for obtaining electrolyzed water according to the invention.
• the advantage and the flexibility of use of the electrolysis module according to the invention makes it possible to obtain, from an abundant source which is water, a less expensive treatment since it does not require conventional chemicals in the process. treatment of diseases or disorders linked to senescence, while avoiding the side effects of said chemicals.
• Electrolyzed water obtained by the process according to the invention, can also be present in more conventional compositions that are found for sale in pharmacies or drugstores or even supermarkets, said composition comprising 95 to 100% electrolyzed water and 0 to 5% of an excipient and / or emulsifier.
• Said composition is for use as a medicament in the treatment of diseases or disorders associated with cellular senescence, more particularly cancers related to age or neurodegenerative diseases.
• It can be in the form of a cream, a gel, a dressing, a patch, for example when it comes to eliminating senescent cells from the skin and maintaining the skin. healthiest skin possible with a maximum of healthy or non-senescent cells.
• Another case which can be considered more particularly is that of grafts of skin cells, tissues or even organ such as for example the liver, is the use of electrolyzed water according to the invention in a culture medium. cells, tissue or organ in vitro.
• This culture medium most often comprises, in addition to the electrolyzed water according to the invention, all the nutrients essential for maintaining its activity such as growth factors, nutrients such as amino acids, salts, and possibly additives. animal serum and other compounds such as sugars or even antibiotics in order to avoid contamination of the culture media, as well as others well known to those skilled in the art of culture media.
• Another potential galenic form for the composition according to the invention can be an aqueous gel also called a hydrogel.
• a hydrogel is a gel in which the swelling agent is water electrolyzed according to the process of the invention.
• the matrix of a hydrogel is generally a network of polymers which are insoluble in water, but are able to swell substantially in the presence of a large amount of water or aqueous solutions.
• FIG. 1 shows the results obtained during comparative tests between a water according to the WDW invention, obtained according to the implementation of the method according to the invention under a current of 62.5 mAh / L, and a water ultrapure MQW which is obtained with filters from the company Millipore, in the presence of variable concentrations of H202, the two waters being applied to cultures of chondrosarcoma cells of the JJ012 cell line.
• the cell culture medium is also of the EMEM type obtained by mixing powders (Thermo Fisher, GIBCO # 41500-034, Penicylin Steptomycin (in English) (PenStrep) # 15140122, and 10% Fêtai Bovine Serum (in English) (FBS) ) # 26140079) and sterilized and filtered (0.2 ⁇ m filter) and containing said cells.
• Senescence-associated b-Gal is an isoform of b-Gal which exhibits optimal activity at pH 6.0 and which is primarily used as a biomarker for senescent cells (K802).
• the detection is based on the hydrolysis by beta-galactosidase of a non-fluorescent substrate to generate a strongly fluorescent product.
• the electrolyzed water obtained by implementing the method according to the invention (WDW) is the most effective in terms of increasing cell senescence than other electrolyzed or commercial water and this when the concentration of H202 is of 0 mM corresponding to the control.

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• 2019
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