EP3313414A1

EP3313414A1 - Kit for treating sepsis and/or any systemic (sirs) or damaging cellular hyperinflammation

Info

Publication number: EP3313414A1
Application number: EP16742342.5A
Authority: EP
Inventors: Vincent COTEREAU
Original assignee: Serenite-Forceville
Current assignee: Serenite-Forceville
Priority date: 2015-06-25
Filing date: 2016-06-24
Publication date: 2018-05-02
Also published as: WO2016207577A1; US11224633B2; US20180185443A1; FR3037798A1; CA3028101A1

Abstract

The present invention relates to a kit that includes a first container, containing a composition including at least one antioxidant seleno-protein, and at least one second container, containing at least one composition including at least one oxidant seleno-compound. The present invention also relates to an administration method that makes it possible to administer effective cytotoxic doses of seleno-compounds, thus making it possible to inhibit hyperactivation of phagocytes, in particular of immature circulating neutrophilic polynuclear phagocytes, and directly and indirectly protects endothelial cells, in particular for treating sepsis, SIRS, and leukemia. Finally, another aspect of the invention relates to an administration device that is suited to the administration method according to the invention.

Description

KIT FOR THE TREATMENT OF SEPSIS AND / OR ANY GENERALIZED HYPER-INFLAMMATION (SIRS) OR CELLULAR

HARMFUL

FIELD OF THE INVENTION

The present invention relates in particular to the treatment of sepsis, systemic inflammatory response syndrome (SIRS) and more particularly to septic shock and SIRS conditions giving similar tables requiring the use of catecholamines after filling or when fast filling phase in case of very sudden aggravation. The present invention also relates to leukemia treatment (for circulating elements). In particular, the present invention relates to a kit combination of a selenium protein and a selenium compound for the treatment of these syndromes and diseases. The present invention also relates to a method of administration which makes it possible to administer effective doses of selenium compounds that are toxic to circulating hyperactive cells and pathogens (bacteria, viruses, fungus and parasites) but beneficial in total for the whole of the organization. Finally, in another aspect, the invention relates to a delivery device.

STATE OF THE ART

Sepsis is an excessive generalized inflammatory response of the body in response to a bacterial infection, most often, but also parasitic, viral or fungal. In the current state of the art, sepsis is a syndrome and not a disease. Severe sepsis is defined by the presence of one or more organ dysfunctions or failures and signs of peripheral hypofusion, whereas septic shock is defined by the presence of persistent hypotension despite adequate filling associated with the patient. more often to multi-visceral failures. Adequate is generally a filling of 20 to 30 mg / kg. In both cases, an answer excessive or inappropriate body complex, requires the implementation of substitutions of major functions in intensive care to avoid death. This complex deployment of large function audiences allows people who die to survive sepsis; however, despite these expensive additions, the mortality rates of these patients, 28 days after the beginning of the episode, are approximately 20 to 30% and 40 to 50% respectively. In addition, the number of resuscitation beds available to treat them effectively is limited. Sepsis causes a very large number of deaths, slightly higher than those caused by myocardial infarction, and is the leading cause of death in hospital and accounts for more than 5% of hospital expenditures (0.5% of GDP). ). It is one of the leading causes of death in the world and would be the first in case of a pandemic where resuscitation structures would be completely overwhelmed. It is one of the leading causes of health spending in the world in current practice. Moreover, it would be a major cause of mortality in the event of a pandemic situation - sepsis is at the origin of deaths in the case of pest epidemics for example -, disaster (associated with crushing syndromes), bioterrorism or war. In the latter case, it will be readily associated with another cause of generalized inflammation that is extensive burns that would account for more than 30% of victims. Acute inflammation is also a major event for radiation victims who do not die immediately.

Currently, the treatment of this medical emergency consists in the administration within one hour of antibiotics, associated with a filling, an oxygenation, the introduction of catecholamines and the care in intensive care in the absence of treatment specific. In addition, the patient may be given low dose corticosteroids, but this remains controversial and has limited effectiveness (if any). Recommendations have been established globally as part of the surviving sepsis campaign, a global collaborative effort involving major resuscitation companies, including US and European. Nevertheless, there is currently no specific treatment for this excessive defense reaction of the body and most of the proposed treatment is to try to allow the body to survive this situation of self. -aggression by the most judicious placement possible of substitution pending the end of this reaction, while having treated the origin of what triggered it (activation of toll-receptors) infectious in the context of sepsis (PAMPS), cellular damage in the context of acute inflammatory syndromes generalized (DAMPS). At the level of the microcirculation, the interaction of phagocytic cells and endothelium, following the very rapid alerting of the cells of the innate secondary response to Toll receptor activation, participates in the modification of the endothelium (flip- flap) from a non-adherent, non-inflammatory and non-coagulating endothelium to an adherent, inflammatory and coagulating endothelium and then injured with abrasion of the endothelium surface layer (ESL), formation of nitro-oxygenated reactive derivative - such as peroxynitrite by the joint action of the polynuclear adherents (emission of anion surperoxide) and activated endothelial cells (emission of NO), emission of microparticles, and opening of the tight junctions and suffering of the endothelium. This dysfunction of the microcirculation and this endothelial pain are considered as one or the major element in the occurrence of visceral failures secondary to severe sepsis and septic shock. These visceral failures can lead to death despite resuscitation in intensive care that allows subjects who should have died to survive transiently or not, and lead to death in the absence of such resuscitation care. It has been possible to consider (mathematical modeling applied to biology) that this situation consisted of a situation of chaos.

The objective of the resuscitation is then to bring the subject, once the acute episode passed, to the attractor of homeostasis by gradually releasing survivors of survival related to locums. This hyper-inflammatory state most often leads to an acquired immune defect that will cause new infections, possibly with non-dangerous pathogens in a non-immunocompromised subject who may again cause a sepsis reaction. The evolution is likely to be then septic shock in septic shock to exhaustion of the subject in a table of marasmus and major undernutrition leading most often to a Therapeutic limitation to an outdated situation where the return to a situation without substitute does not seem possible anymore and corresponds to relentlessness.

The reactive derivatives of oxygen (polynuclear respiratory oxidation (oxidative metabolism), endothelial cell hyperactivation with NO synthesis, ischemia-reperfusion phenomena and mitochondrial pain) generated by the fight against infectious agents play a major role in endothelial dysfunction, the first step towards multi-visceral failures and death. Selenium is a divalent atom in physiopathology. On the physicochemical level, it belongs to column VI which is that of oxygen (chemical family of chalcogens). In fact, chemically and biochemically, small selenium compounds are often toxic oxidizing molecules.

Thus, sodium selenite is one of the most toxic selenium compounds and its toxicity is similar to that of arsenic salts. A concentration of 10 μιηοΙ / L results in cell killing for detached cells (Stewart MS, Free Radic Biol Med 1999 Jan; 26 (1-2): 42-8). A selenium concentration of 18 μιηοΙ / L is observed in cases of fatal intoxication (Nuttall KL, Ann Clin Lab Sci 2006 Autumn, 36 (4): 409-20). The lethal dose in humans appears to be in the range of 1.5 to 3 mg / kg. Sodium selenite will react preferentially with the disulfide bridges. Its toxicity is greater on free cells than on cellular carpets, and also much more important on hyper-activated cells than on quiescent cells. In vitro, its cytotoxic activity has been demonstrated particularly on cancer cells for concentrations above 2 to 5 μιηοΙ / L of selenium in the form of sodium selenite. The toxicity of sodium selenite seems to be mainly at the level of the mitochondria, particularly in hyperactivity cells.

Selenium is incorporated in selenium-like proteins, and is included in the molecule Sodium Selenite, allowing it to be a key element of antioxidant defense in mammals. As a result, cases of acute poisoning by selenium compounds are completely reversible when they are not fatal, selenium once incorporated into these selenium-containing proteins with an antioxidant action. opposite to that of selenious oxidizing compounds such as selenite. As part of the improvement of their antioxidant defense (and their regulation of intra-cellular redox potential modulating all cellular activity), mammals and birds have acquired a particular amino acid, selenocysteine, where selenium replace the sulfur in cysteine. This replacement is genetically controlled, with the modification of the meaning of the UGA stop codon due to the presence of a neighboring structure and requires energy in the form of ATP. The presence of this amino acid at the active site of the selenated enzymes will allow them to have an antioxidant action faster and more robust than their iron precursor and the replacement of selenocysteine by cysteine at the level of active site decreases by a factor of 1000 the effectiveness of these enzymes. Selenated proteins are major antioxidant enzymes in mammals. Selenium deficiency is fatal in humans and the knockout model for selenocysteine is lethal.

At the plasma level, two selenic proteins are observed, selenoprotein-P, a protein that is highly conserved among species and exceptionally among selenium-containing enzymes because it contains 9 amino acids selenocysteine (while the others have only one amino acid). selenocysteine) and plasma glutathione peroxidase. Selenoprotein-P (Sel-P) accounts for 60% of plasma selenium. It transports biologically active selenium (selenocysteine) between the liver and the tissues and directly contributes to the antioxidant protection of the endothelium by an enzymatic action, likely detoxifying, of the peroxynitrite ion. It has been shown to have a membrane stabilizing role limiting the emission of microparticles.

Due to the understanding of the toxicity of sodium selenite, this selenite has been administered continuously or with 30-minute boluses as described in Angstwurm MW (Crit Care Med., 2007 Jan. 35 ( l): 118-26), which shows a decreasing trend in mortality with an administration of 1000 μg of selenium in the form of sodium selenite (14 μg / kg) followed by a continuous dose of the same dose. 24 hours, continued for 15 days, but whose results have not been reproduced at present (Angstwurm MW, Crit Care Med 2007 Jan, 35 (l): 118-26). The maximum dose of 1000 μg / day is recommended by Heyland DK (Crit Care 2007; 11 (4): 153), a dose of 4000 μg / day (55 μg / kg) administered continuously having had no beneficial effect because in its view of negative effects. The administration of 2000 μg (80 μg / kg) bolus in sheep was accompanied by positive effects, in contrast to the administration of the same dose continuously, however, it was not possible to conclude on the interest of a bolus administration (Wang Z et al., Shock 2009 Aug; 32 (2): 140-6). In fact, the interest of this bolus administration remains very controversial (Alhazzani, W. et al Crit Care Med 2013 Jun; 41 (6): 1555-64). Currently, the recommendations of the European Society of Nutrition (ESPEN) are not to exceed 750 to 1000 μg / day because of the risks of toxicity (Singer P et al., Clinique Nutr., 2009 Aug; 28 (4): 387-400 ), this corresponds to the NOAEL No Adverse Ejfect Level determined in nutrition (800 μg, ie 11 μg / kg and per day). As for the American nutrition company ASPEN, it recommends an optimum intake between 500 and 750 μg / day. In view of the different results of the studies, the benefit of selenium administration in sepsis resuscitation patients is not recommended in Dellinger RP et al. (Crit Care Med 2013 Feb; 41 (2): 580-637) and Casaer MP et al. (N Engl J Med 2014 Mar 27; 370 (13): 1227-36).

However, the Applicant has remained convinced that the administration of compounds containing selenium (or selenite) is a suitable response to manage circulating toxic elements in the context of sepsis and leukemia. In this way, the Applicant has questioned how to optimize the administration of compounds containing selenium, while protecting the body from their toxic effects. To this day, and to his knowledge, this technical problem is not resolved. And in any case, there is a very strong need for means to significantly reduce the high mortality rate in subjects with sepsis and leukemia.

ABSTRACT

The object of the invention is to propose a solution to this technical problem, and proposes a treatment that targets phagocytes, in particular neutrophils. by means of a two-stage administration of compounds, preferably selenic, of different natures, at first an antioxidant protein, preferably a selenium protein, then in a second time, an oxidizing compound, preferably a selenium compound, ultra-fast administration (also called flash administration) , and preferably in a few repetitive flashes, and not in a bolus. The invention directly and indirectly protects the endothelial cells, limits the adhesion of neutrophils to endothelial cells while allowing to inhibit hyper-activation of phagocytes and particularly immature circulating neutrophils. It can also participate in antibiotic therapy, when the subject is subjected to antibiotic therapy, by a cytotoxic effect on circulating pathogens, including when the subject is resistant to antibiotic therapy.

DEFINITIONS

In the present invention, the terms below are defined as follows:

"Treatment", "treat" or "relieve" are terms related to therapeutic treatment, and mean delaying the onset of a syndrome or illness, stopping the syndrome or illness, or reversing the syndrome or illness . In one embodiment, the syndrome is sepsis; the subjects concerned by the invention therefore include subjects at the initial phase of sepsis, diagnosed in particular by a decrease in circulating selenoprotein-P or by any other biomarker of sepsis, and / or subjects in whom the sepsis is already diagnosed or recognized. A subject is considered to be effectively treated with sepsis if he or she shows an observable and / or measurable decrease in nitrogen oxide and / or lactate production, and / or a significant decrease in the mortality rate as well as a decreased severity of visceral failures (which can be measured by the SOFA score). These evaluation parameters of an effective treatment are easily measurable by routine procedures known to the physician. In another embodiment, the disease is cancer, circulating or solid tumor; the subjects concerned by the invention therefore include subjects suffering from cancer.

In the present invention, an "excipient" means any substance other than the active ingredient present in a composition which confers on it stability properties. of form (liquid, solid, capsule, etc. depending on the mode of administration), of taste, of dissolution (for example targeted dissolution in the stomach or the digestive tract), of color, etc. A "pharmaceutically acceptable excipient "acceptable" more specifically means an excipient which does not induce an allergic or undesired reaction when it is administered. This definition includes in particular all pharmaceutically acceptable solvents and dispersion media.

"Effective amount" (or "therapeutically effective amount") for the treatment of sepsis refers to an amount necessary or sufficient to, without causing significant and adverse side effects to the subject, (1) delay or stop sepsis, (2) make improvements to sepsis, (3) reduce the severity or incidence of sepsis, (4) stop or treat sepsis; with regard to the administration of the antioxidant protein, preferably selenium, administered first, according to the invention, the effective amount is that capable of protecting the endothelium against inflammation and hyperoxidation induced by the sepsis, but also against the toxicity of sodium selenite or of another selenious oxidizing compound that can be administered later; an effective amount of antioxidant protein, preferably selenium, may be administered prior to the onset of sepsis, for prophylactic or preventive action; alternatively or additionally, an effective amount of selenium protein may be administered after the onset of sepsis for therapeutic purposes; as regards the selenium compound, the effective amount of selenium compound should be administered only after administration of the effective amount of antioxidant, preferably protective, selenium protein, and is between 50 and 600 μg / kg d selenium equivalent administered at a cytotoxic concentration so as to reach a plasma concentration of the oxidizing selenium compound of about 2 to 40 μιηοΙ / L, preferably 10 to 20 μιηοΙ / L. the effective amount of selenium compound is that necessary or sufficient to reduce the hyper-activation of phagocytic cells or polynuclear neutrophils, and control Γ hyper-inflammation, the state of hyper-oxidation and stop the chaos situation by a cytotoxic action targeted and transient. "Super-fast" or "flash" administration according to the invention refers to administration at a rate of more than 2 to 15 mL / sec, preferably 3 to 10 mL / sec, more preferably 4 to 5 mL / sec.

Administration "in repeated flash" or "split" corresponds to the succession in the short term, about 5 to 20 minutes, preferably about 10 minutes, from one to 10, preferably 2 to 5, very preferably 3 administrations in flash.

The term "slow administration" according to the invention refers to an amount of selenium protein administered at a rate of about one quarter to two or five times the total selenoprotein-P of the subject. The determination of the amount of total salt-P is in particular described in the European patent: EP1393078 or US Pat. No. 7,635,491. The rate of administration is calculated so that administration lasts from about 10 minutes to one hour, preferably about 30 minutes. For example, for a man of 70 kg with a plasma volume of about 3000 ml, about 2 to 90 mg of selenoprotein-P in 10 min to one hour and preferably in 30 minutes, a speed of 2 to 540 mg / hour, preferably 50 to 400 mg / hour, very preferably 80 to 200 mg / hour. The amount of total selenoprotein-P varying according to the subjects and the practitioner will know how to determine the rate of administration adapted to each. This injection can be repeated one to three times a day, especially the first day every 4, 6 or 12 hours (one to four administrations / day of treatment).

"About", placed in front of a number, means plus or minus 20% of the nominal value of that number.

"subject" in the sense of the present invention refers to a mammal or a bird. The mammal can be a human (patient), nonhuman primate, mouse, rat, dog, cat, horse, or cow, but is not limited to these examples. A subject can be male or female. A subject may be an adult, a child or an infant. A subject is a patient diagnosed with a syndrome, including sepsis, or a disease, including leukemia. A subject according to the invention may also be an organ or a graft. The term "sepsis" refers to the definition of sepsis 2016 and is defined as life-threatening organ dysfunction caused by a deregulated response to an infection. Organ dysfunction can be assessed by increasing the SOFA by 2 points (or more) which is associated with a morality of more than 10%. The term "septic shock" should be defined as subjects in sepsis where there is particularly pronounced circulatory failure, with cellular and metabolic abnormalities that are associated with higher mortality than in sepsis alone (Singer M and JAMA, 2016; 315 (8): 801-810). DETAILED DESCRIPTION

Thus, in a first aspect, the invention relates to a kit comprising: a first container comprising a composition comprising an antioxidant protein, preferably a selenium protein; and at least one second container comprising at least one composition comprising at least one oxidizing compound, preferably a selenium compound.

According to one embodiment, the selenium antioxidant protein is selected from the group comprising or consisting of: selenoprotein P, glutathione peroxidase, thioredoxin reductase, iodothyronine deiodinase, formate dehydrogenase, methionine sulfoxide reductase, selenophosphase synthetase , selenoprotein Pa (SelPa), selenoprotein W (SelW), selenoprotein T (SelT), selenoprotein R (SelR or SelX), selenoprotein 15 kDa (Sell5), selenoprotein N (SelN), selenoprotein T ( SelT2), selenoprotein M (SelM), selenoprotein G-rich (G-rich), selenoprotein W2 (SelW2), selenoprotein BthD (BthD), selenoprotein H (SelH), selenoprotein I (Sell), selenoprotein K (SelK), selenoprotein O (SelO), selenoprotein R (SelR), selenoprotein S (SelS) and selenoprotein Pb (SelPb). According to one embodiment, the composition of the first container comprises a selenium-like protein as described above, in association with any pharmaceutically acceptable excipient. In one embodiment, the selenium antioxidant protein is selenoprotein P in its complete form or in truncated forms, in a 51 kDa isoform or a 61 kDa isoform, of human origin or another mammal or bird, or glutathione peroxidase, including plasma glutathione peroxidase. In one embodiment, the first container further comprises heparin-binding protein (HBP) proteins, preferably antithrombin III (AT III). According to one embodiment, the oxidizing selenium compound is selected from the group consisting of or consisting of: selenite sodium, selenodiglutathion, selenomethyl selenocysteine, dimethyl selinoxide, selenocystamine, or chemical synthesis derivatives containing one or more selenium atoms, selenium hybrid, fluorinated selenium salt, chlorinated selenium salt, brominated selenium salt, iodinated selenium salt, selenoxide, selenium salt, selenium salt, selenium salt, potassium selenium salt, selenium salt of germanium, salt of barium selenium, selenium salt of lead, selenium salt of zinc or a selenium salt of nitrogen, atonimy (III), selenide (Sb ₂ Se ₃ ), arsenic (III), selenide (AS ₂ Se ₃ ), bismuth ( III) selenide (Bi ₂ Se ₃ ), cadmium selenide (CdSe), cobalt (II) selenide (CoSe), mercury (II) (HgSe), selenium oxychloride, selenyl chloride (Cl ₂ OSe), selenium sulfide, s elenium disulfide (SeS ₂ ), silver selenide (I) (Ag ₂ Se), selenide of indium (III) (In ₂ Se), strontium selenide (SeSr), selenic acid (I ^ C ^ Se), selenium dioxide (0 ₂ Se), selenium (Se), selenious acid, selenious acid (H ₂ O ₃ Se), selenoglutathione.

According to another embodiment, the oxidizing selenium compound is a methylated derivative of selenium. In another embodiment, the oxidizing selenium compound is an amino acid containing selenium. According to another embodiment, the oxidized selenium compound is a selenium-containing (selenium-containing) organic compound included in the group of selenium organic compounds comprising but not limited to alkyl, alicyclic, cyclane, terpene, aromatic and heterocyclic compounds. including selenium methionine, selenourea, or selenium diethyledithiocarbamate.

In another embodiment, the oxidizing selenium compound is a hybrid selenium of the formula Se (x) H (y) where x and y are independently an integer of 1 to 10, preferably x is 1 and y is 2. According to one embodiment, the oxidizing selenium compound is sodium selenite (Na ₂ SeO ₃ ), preferably in a non-pentahydrate form. According to one embodiment, the selenium compound is selenocysteine.

According to one embodiment, the composition of the second container comprises an oxidizing selenious compound as described above, in association with any pharmaceutically acceptable excipient.

According to another embodiment, the kit according to the invention comprises a third antioxidant container comprising vitamin C at low dose (antioxidant), vitamin E, a glutathione precursor, iron salts, gold salts, low-dose copper salts (antioxidant), and / or zinc salts.

According to another embodiment, the kit according to the invention comprises a fourth oxidizing container, comprising vitamin C in a high dose (oxidizing dose, greater than 20 mg), iron salts, gold salts, salts and salts. high dose copper and / or tellurized compounds.

In one embodiment, the kit according to the invention comprises:

at. a first container, comprising a composition comprising an antioxidant protein, preferably selenium;

b. one, two or at least three second containers, each of the second containers comprising at least one composition comprising at least one oxidizing compound, preferably selenium, preferably sodium selenite or selenocysteine; in one embodiment, all the second containers comprise the same composition;

vs. optionally at least one third container of oxidizing compounds comprising vitamin C, vitamin E, a glutathione precursor, zinc salts, arsenic salts, copper salts (in low concentration) and / or proteins heparin-binding protein (HBP) group, preferably vitamin E;

d. optionally at least one fourth container of antioxidant compounds comprising vitamin C in high concentration (greater than 20 mg / L), iron salts, gold salts, copper salts, and / or tellurium compounds.

According to one embodiment, a container is a dosage unit. Preferably, the first container containing an antioxidant protein, preferably selenium, has a capacity of 5 to 100 ml, preferably 20 to 50 ml. Preferably, the second container containing the selenium compound has a capacity of 0.05 to 10 mL, preferably 4 mL.

In one embodiment, the composition included in the first or second container includes at least one pharmaceutically acceptable excipient. The pharmaceutically acceptable excipients according to the invention comprise water, physiological saline, Ringer's solution, a dextrose solution and ethanol solutions, glucose, sucrose, dextran, mannose, mannitol, sorbitol , polyethylene glycol (PEG), phosphate, acetate, gelatin, collagen oils, Carbopol®, vegetables, and the like. Also suitable preservatives, stabilizers, antioxidants, antimicrobial agents and buffering agents, such as, for example, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), citric acid, tetracycline can be included. , and the like.

Other examples of pharmaceutically acceptable excipients which can be used in the invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffering substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulphate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene polyoxypropylene block polymers, polyethylene glycol and wool grease.

Other examples of pharmaceutically acceptable excipients which can be used in the invention include, but are not limited to, surfactants (eg hydroxypropylcellulose); suitable carriers, such as, for example, solvents and dispersion media containing, for example, water, ethanol, a polyol (eg glycerol, propylene glycol and liquid polyethylene glycol, and the like), mixtures suitable thereof, and vegetable oils, such as, for example, peanut oil and sesame oil; isotonic agents, such as, for example, sugars or sodium chloride; coating agents, such as, for example, lecithin; agents delaying absorption, such as, for example, aluminum monostearate or gelatin; preservatives, such as, for example, benzalkonium chloride, benzethonium chloride, chlorobutanol, thimerosal and the like; buffers, such as, for example, boric acid, sodium or potassium bicarbonate, sodium or potassium borates, sodium or potassium carbonates, sodium acetate, sodium bisphosphate and the like ; tonicity agents, such as, for example, dextran 40, dextran 70, dextrose, glycerin, potassium chloride, propylene glycol, sodium chloride; antioxidants and stabilizers, such as, for example, sodium bisulfite, sodium metabisulfite, sodium thiosulfite, thiocarbamide and the like; nonionic wetting agents or clarifying agents, such as, for example, polysorbate 80, polysorbate 20, poloxamer 282 and tyloxapol; viscosity modifiers, such as, for example, hydroxyethylcellulose, hydroxmethylpropylcellulose, lanolin, methylcellulose, petrolatum, polyethylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, carboxymethylcellulose; and the like.

According to one embodiment, the compositions included in the kit according to the invention are suitable for administration by injection, such as, for example, by intravenous, intrathecal, intradermal, intramuscular or epidural injection. In one embodiment, the administration is intravenous. In another embodiment, the kit is adapted for intravenous, preferably peripheral, administration. In another embodiment, the kit is adapted for delivery by a central venous catheter (e.g. via the jugular, subclavian or femoral vein), an arterial catheter for local effect or in a chaos situation. According to one embodiment of the invention, the compositions included in the kit of the invention are adapted for administration preferably by a multi-way central catheter so as to avoid reactions with other drugs.

According to another embodiment of the invention, the catheter used is a catheter impregnated with antiseptic, and / or antibiotic. According to one embodiment, the compositions included in the kit according to the invention are suitable for administration by injection into an extracorporeal circulation device.

Examples of formulations suitable for administration by injection include, but are not limited to, an injectable solution or an injectable emulsion (such as, for example, an oil-in-water emulsion, a water-in-oil emulsion, an anhydrous emulsion, an solid emulsion or microemulsion).

In a second aspect, the invention relates to the kit as described above, for its use in a sequential administration method, to a subject in need, said subject being able to be a patient, compositions included in the different containers of the kit. This method is described below.

In a third aspect, the invention relates to a method for the sequential administration of the compositions contained in the kit of the invention, to a subject who needs it, a therapeutically effective amount of the compositions included in the various containers of the kit. described above, in a sequential mode, namely in the first place the administration of an effective quantity of the composition of the first container, namely a composition comprising an antioxidant protein, preferably a selenium-containing protein, continuously, then administering in flash an effective amount of the composition of the second container, namely a composition comprising an oxidizing selenious compound. In one embodiment, the flash administration of the composition of the second container is flash administration repeated one to five times. In one embodiment the flash administration of the composition consists of three successive flash administrations of the composition of the second container. In a fourth aspect, the invention relates to a method for the sequential administration of the compositions contained in the kit of the invention, to a subject who needs it, a therapeutically effective amount of the compositions included in the various containers of the kit. described above, in a sequential mode, namely in the first place the administration of an effective quantity of the composition of the first container, namely a composition comprising an antioxidant protein, preferably a selenium-containing protein, continuously, optionally complemented by the administration of the composition of the third container and then the flash administration of an effective amount of the composition of the second container, optionally supplemented by the administration of the composition of the fourth container. In one embodiment, the flash administration of the composition of the second container is flash administration repeated one to five times. In one embodiment the flash administration of the composition consists of three successive flash administrations of the composition of the second container. According to another embodiment, the method of the invention comprises a preliminary step of determining the opportune moment to start the administration of the compositions contained in the kit of the invention. According to one embodiment, the administration according to the invention is started early before endothelial lesions and after the onset of hyper-inflammation. In another embodiment, determining the appropriate time to commence administration includes detecting and quantifying P-Salt in a subject's plasma. In one embodiment, the start of treatment is decided when the subject shows a plasma decrease from one-half to three-quarters of the reference level of Sal-P, which is about 5 mg / L: in others In other words, if the subject has a plasma concentration of Salt-P of 1 to 2.5 mg / L, the start of treatment can be decided. The determination of the amount of salt P is in particular described in the European patent: EP 1 393 078 or US Pat. No. 7,635,491 incorporated herein by reference. The increase in lactate is also a good indicator for the start of treatment.

According to one embodiment of the invention, the selenium antioxidant protein according to the invention must be or is administered to the subject in an amount likely to make it return to average plasma levels in humans. The man has indeed a plasma concentration of Salt-P of about 5 mg / L. To return to a normal level of plasma Sel-P approaching about 5 mg / L in a subject with sepsis or acute generalized inflammation, for example, the amount of selenoprotein-P administered is, in one embodiment from 2 to 90 mg, preferably 2 to 30 mg, very preferably 4 to 30 mg of selenoprotein-P administered slowly, that is to say for about 10 minutes to about one hour and preferably about 30 minutes. According to another embodiment of the invention, the selenium protein according to the invention must be or is administered to the subject 1, 2, 3, 4, 5 or 6 times daily for 1, 2, 3 or 4 days, preferably 3 days.

According to another embodiment, the selenium compound must be or is administered in flash so as to obtain a concentration of the selenium element in the plasma of about 2 to 40 μιηοΙ / L, preferably 10 to 20 μιηοΙ / L. According to one embodiment, the flash administration is the injection of a composition of selenium compound, preferably selenite sodium or selenocysteine, concentration of 0.1 mg / ml to 10 mg / ml, preferably 0.2 at 5 mg / mL or in another embodiment of about 1.5 mg / mL to 30 mg / mL, preferably 5 mg / mL to 15 mg / mL, expressed as selenium equivalent, in a time of about 0.1 to 4 seconds, preferably about 0.5 to 1 second. In a particular embodiment, the concentration of selenium equivalent administered flash is from about 0.05 to about 0.6 mg / kg body weight, preferably 0.05 to about 0.3 mg / kg body weight, very preferably from about 0.1 to 1 mg / kg, from about 0.2 to 0.5 mg / kg, preferably from about 0.25 to 0.35 mg / kg, preferably from about 0 to , 23 mg / kg.

The injection rate is preferably about 1 to 10 mL / sec or 2 to 10 mL / sec, preferably about 3 to 4 mL / sec or 4 to 5 mL / sec. Administration may be by central catheter or intravenous administration.

According to one embodiment, the therapeutic scheme extends over several days, preferably 1, 2, 3 or 4 days and the composition comprising the antioxidant protein, preferably selenium, is administered every 4, 6, 8 or 12 hours, continuously for 10 minutes to 1 hour, said administration being followed, between 10 minutes to two hours (preferably 10 to 20 minutes) after the end of the administration of the selenium protein of a three flash composition comprising the oxidizing compound selenie, the flash being made 5 to 20 minutes apart. According to one embodiment, the composition comprising the antioxidant protein, preferably selenium, is administered for 30 minutes, then after a waiting time of 10 minutes, the composition comprising the selenium compound is administered in three spaced flash injections of 10 minutes; the whole is repeated after 4, 6, 8 and / or 12h.

In one embodiment, the content of the third container (preferably Vitamin E) is administered continuously before administration of the flashes of the composition comprising the selenium compound (s), preferably over 10 minutes to 2 hours, preferably during administering the composition of the selenium protein composition.

In one embodiment, the contents of the fourth container (preferably, high concentration Vitamin C) are administered continuously beginning within the flash time of selenium compounds, and preferably for 10 minutes at 2h.

According to one embodiment of the invention, the administration may be repeated, with iterative injections followed by rinsing with a neutral solution type saline or glucose, according to the terms of what is achieved for the injections of contrast product during scanners. This modality of administration with flash injections makes it possible to obtain cytotoxic concentrations, particularly on hyper-activated inflammatory cells, neutrophils and thus the therapeutic effect by limiting the quantity of product administered and therefore its toxicity, in particular pulmonary and / or or heart.

Flash administration mimicking a rapid injector of the type used in imaging was chosen to optimize the peak cytotoxic concentration of the selenium compound according to the invention. In addition, the repetition of the doses, at a short interval or fractional administration, made it possible to optimize the cytotoxic effect on hyperactivated phagocytic cells and the prior administration of selenoprotein-P to protect the endothelium. Moreover, the dose administered in total on the three injections in flash is much higher than those administered until now. The divided dose allows thus to obtain very high transient and repeated peaks of the active product while limiting the risk of lung and / or cardiac toxicity.

In one embodiment, the oxidizing selenium compound of the second container is included in a transport or vectorized means for targeting phagocytic cells or the innate response. According to one embodiment, the oxidizing selenium compound is vectorized or included in a transport means to target in particular immature circulating neutrophil polynuclear cells. According to another embodiment, the oxidized selenium compound included in a means of transport or vectorized according to the invention expresses specific receptors for phagocytic cells or the innate response such as, for example, the CD64, CD36 receptor. According to another embodiment, the oxidized selenium compound included in a transport means or vectorized express receptors specific immature circulating neutrophils or cancer cells or bacteria or viruses or parasites or fungal agents.

According to another embodiment, the means of transport according to the invention is a ghost red blood cell. The preparation of ghost red blood cells is described in Dodge JT et al. Arch Biochem Biophys. 1963 Jan; 100: 119-30; Shaillender M. et al. International Journal of Pharmaceutics 415 (2011) 211-217. The ghost red blood cell is used as a means of transporting and presenting the selenium compound; it makes it possible to avoid the systemic side effects of selenious oxidizing compounds whose severity (lethal) is known; it also makes it possible to act at very high concentrations of the selenium compound at the level of the target cells, in this case the phagocytic cells and more particularly the immature circulating neutrophilic neutrophils at the level of the microcirculation and to protect directly and indirectly the endothelium while at the same time decreasing hyperactivation, cancer cells, bacteria, viruses or parasites or fungal agents.

Within the targeted cell, the preferred targets are the mitochondria to reduce the activation of hyperactivated polynuclear cells or other hyperactive cells or the nucleus so as to block the binding of NFKB to the DNA and thus to limit or block the generalized inflammatory reaction. According to another embodiment, the means of transport or vector according to the invention is a nanoparticle. Examples of nanoparticles according to the invention are described in the article by Vergaro V. et al. Drug-loaded polyelectrolyte microcapsules for sustained targeting of cancer cells. Adv Drug Deliv Rev. 2011 Aug 14; 63 (9): 847-64. For example, the nanoparticles according to the invention include but are not limited to: a solid lipid nanoparticle, a nanostructured lipid transporter, a dendrimer, a magnetic nanoparticle, a fullerene, a carbon nanotube, a halloysite nanotube, a colloid, a colloid coated with polyelectrolytes, a nanocolloid, polyelectrolyte microcapsules. According to another embodiment, the means of transport or vector according to the invention is a capsule. Preferably, the capsule according to the invention is a capsular bag.

According to one embodiment, the means of transport or vector according to the invention is a liposome. According to one embodiment, the vector according to the invention is a cationic liposome. Liposomes known to those skilled in the art include but are not limited to RPR209120 / DOPE. According to another embodiment, the vector according to the invention is a mannosylated cationic liposome to specifically target neutrophils. Liposomes for targeting neutrophils are described, for example, in Kelly C. et al. (Targeted liposomal drug delivery to monocytes and macrophages, J Drug Deliv 2011, 2011: 727241). Formulations for targeting phagocytes are well known to those skilled in the art.

According to another embodiment, the subject of the invention is suffering from cancer, in particular leukemia, lymphoma or solid tumor cancer. In a particular embodiment, the subject is suffering from leukemia or lymphoma. Examples of leukemias include but are not limited to: acute leukemia, chronic leukemia, lymphoid lineage leukemia, myeloid lineage leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, acute myeloblastic leukemia, chronic myeloid leukemia. Examples of lymphomas include but are not limited to: lymphoma, Hodgkin's and non-Hodgkin's lymphoma. According to one embodiment, the subject is suffering from a solid tumor. Solid tumors include carcinomas and sarcomas. Examples of carcinomas include but are not limited to: cancers of the breast, lungs, prostate, intestine. Examples of sarcomas include but are not limited to: bone cancers, cartilage.

The present invention also relates to a device adapted to a biphasic mode of administration of the kit according to the invention.

According to one embodiment, the device comprises a flash administration mode.

According to another embodiment, the device comprises a slow mode of administration.

The delivery device can not be performed with the usual infusion pumps and requires an injection with a very high pressure level up to 8 bar and allowing injection up to 10 mL / sec. In large mammals and especially in humans, the device comprises an automated high pressure pump of the type used in imaging allowing very high flow rates, especially in central catheters.

According to another embodiment, the device comprises an extracorporeal circulation device. Preferably, this extracorporeal circulation device allows an attack of the figured elements of the blood without toxicity to the endothelium. According to one embodiment, the device according to the invention provides a source of energy for generating a pressure and / or a fluidic flow, while offering the user a tactile and / or audible feedback of the fluidic pressure generated, which allows the user to modulate the pressure and / or flow of the fluid. The injection system of the present invention is capable of providing both a low flow rate and / or low pressure delivery mode as well as a high flow and / or high pressure delivery mode. According to one embodiment, the injection system of the device according to the invention comprises a fluid control module in active connection with an injector connected to a syringe. The syringe is in fluid connection with an automatic valve of the fluid control module, which is also in fluid communication with a contrast source through an intermediate flow chamber. The dropper chamber preferably comprises a fluid level detection mechanism. A preferably automated valve is also in communication with a first lumen inlet of a pressure isolation valve. The valve prevents saline and / or contaminated fluids from entering the syringe and allows the control module to stop the flow of fluid from the syringe quickly at any pressure and / or flow rate. This ability to immediately stop the flow of injection fluid at any pressure and flow rate substantially eliminates the effects of system compliance and allows for both slow administration of a compound and both flash administration of another compound.

The invention also relates to a kit as described above, for its use in the treatment of sepsis and / or systemic inflammatory response syndrome (SIRS), severe sepsis, septic shock, cancer (in particular leukemia, lymphoma or solid tumor cancer), said use implementing the method of administration described above. In one embodiment, the treated subject may be in septic shock. In one embodiment, the treated subject may be in a multi-visceral failure state.

The present invention relates in particular to a kit for its use in the treatment of sepsis and / or systemic inflammatory response syndrome (SIRS) comprising on the one hand an antioxidant selenium protein, and on the other hand a compound selenium, the selenium antioxidant protein being intended to be administered slowly before administration of the selenium compound, which is to be administered after the selenium protein ultra-fast, preferably repeated flash. The terms sepsis and SIRS as used in the invention may correspond to septic shock and SIRS conditions giving similar tables requiring the use of catecholamines after filling or during rapid filling phase in case of aggravation very brutal. Severe sepsis may be indicated by the presence of one or more organ dysfunctions or failures and signs of peripheral hypofusion. Severe sepsis is an inflammatory systemic response to an infection associated with visceral failure, hypo-infusion or hypotension, including lactic acidosis, oliguria, or altered mental status. Septic shock can be revealed by the presence of persistent hypotension despite adequate filling. Adequate is generally a filling of 20 to 30 mg / kg. Septic shock is often characterized by an inflammatory systemic response to an infection associated with collapse (hypotension) of more than one hour despite adequate filling and in the absence of other causes of hypotension. (Hypotension is later in children and other signs should be considered in pediatrics). Hypotension is defined as: systolic blood pressure <90 mmHg, mean arterial pressure <60 mmHg, or decrease> 40 mmHg in adults or +/- 2 SD below that of reference in children).

Severe sepsis, like septic shock, is a syndrome based on the clinical reflection of visceral failure (s), itself (s) consequence of the metabolic response to aggression.

The inflammatory systemic response (SIRS) is a descriptive term used to describe the presence of a generalized inflammatory response regardless of its cause, and is recognized by a complex set of signs. According to the latest consensus conference, according to the "2001 SCCM / ESICM / ACCP / ATS / SIS International Sepsis Conference Definitions", these signs are currently defined as follows:

• General settings :

- fever or hypothermia (<36 or> 38.3 ° C),

- Tachycardia> 90 bpm (beats per minutes) or +/- 2 standard deviation (SD) compared to the reference value for age,

- Tachypnea> 30 bpm, - Mental status disorder,

- Edema or positive hydrosodium balance (> 20 ml / kg over 24 hours).

Inflammatory parameters:

- Hyperleukocytosis or leukopenia (> 12,000 or <4000 / μ1),

- Presence of> 10% of immature forms,

- C reactive plasma protein (CRP) +/- 2 DS reference values,

- Plasma procalcitonin (PCT) +/- 3 DS reference values.

Hemodynamic parameters:

- Hypotension (systolic blood pressure <90 mmHg, mean arterial pressure,

- <70 mmHg, or decrease> 40 mmHg in adults or +/- 2 SD below that of reference in children).

- Oxygen saturation of mixed venous blood (Sv0 ₂ )> 70% (not usable in children),

- Cardiac index> 3.5 L / min.m (not usable in children).

Organ dysfunction:

Arterial hypoxemia (Pa0 ₂ / Fi0 ₂ <300),

- Oliguria (acute) (diuresis <0.5 ml / kg / h or 45 ml for the last 2 hours),

- Increased serum creatinine> 0.5 mg / dl,

- Coagulation abnormalities (activated thromboplastin time> 60 s or> 1.5 normalized international percentage),

Ileus (abdominal silence on auscultation),

- Trombocytopenia <100,000 / μ1,

- Hyperbilirubinemia (total bilirubinemia> 4 mg / dl or 70 mmol / L).

Tissue perfusion parameters:

- Hyperlactatemia> 3 mmol / L,

- Increased capillary reperfusion time, The term multi-visceral failure as used in the invention corresponds to a clinical syndrome characterized by an acute and potentially reversible dysfunction of an organ or a large function, not directly involved in the initial pathological process. Multi-visceral failure is usually associated with hypotension (systolic blood pressure <90 mmHg or reduction> 40 mmHg from baseline in the absence of other cause).

The multi-visceral failures are readily based on the SOFA (Sepsis Related Organ Failure Assessment) score (Table 1), the first two ranks are associated with dysfunctions, the last two with failures. They then associate themselves most often with the need to implement locum to keep the subject alive. This may include (i) ventilating by a device to assist breathing, (ii) infusion and catecholamine administration to maintain satisfactory blood pressure and blood flow, (iii) dialysis to replace the kidneys, (iv) transfusion of blood and platelets. These substitutions, and their supervision, will have a very important impact in the cost of treating subjects in septic shock.

Table 1 (SOFA score from 0 to 20) (Dopa for Dopamine, Adré for Adrenaline, Noradré for Noradrenaline, diu for diuresis in ml / d.) The score of SOFA (Sepsis Related Organ Failure Assessment) is one of the failure scores the most used. The definition of sepsis was changed in 2016 and is defined as life-threatening organ dysfunction caused by a deregulated response to infection. The Organ dysfunction can be assessed by increasing the SOFA by 2 points (or more) that is associated with a morality of more than 10%. Septic shock should be defined as subjects in sepsis where there is particularly pronounced circulatory failure, with cellular and metabolic abnormalities that are associated with higher mortality than in sepsis alone (Singer M and JAMA 2016; 315 (8 ): 801-810).

Markers for monitoring the evolution of sepsis are known to those skilled in the art. Lactate plasma concentration and its evolution is a late but commonly accepted marker of the occurrence of sepsis and the monitoring of the efficacy of the management of septic shock (or severe sepsis). other recognized markers are those of visceral failure surveillance as listed in the 2001 consensus conference and in the SOFA score. Other markers such as nitrogen oxide NO, cytokines, growth factors in particular endothelium, microparticles, receptor assays are used but in the context of research. The determination of selenoprotein-P has been proposed as an early marker of sepsis and its severity.

According to one embodiment, the subject of the invention is a subject suffering from sepsis and / or acute generalized inflammation.

According to another embodiment, the subject of the invention is suffering from early phase sepsis.

According to one embodiment, the subject of the invention is a subject having undergone cardiac arrest or ischemia reperfusion.

According to one embodiment, the subject of the invention is under antibiotic treatment or is suffering from a bacterial infection particularly resistant to antibiotics or is suffering from a viral infection.

According to one embodiment, the subject of the invention is an organ and the method according to the invention is implemented before organ removal, within the framework of a graft preparation, as part of a transplant and / or as part of transplant rejection.

Pre-existing conditions may make the subject more likely to develop sepsis (as defined by the document, severe sepsis and septic shock) following an infection: alcoholic liver disease, cirrhosis, anorexia, undernutrition, malnutrition, diabetes, cancer, hemopathy, systemic disease, a subject with Acquired Immunodeficiency Syndrome (AIDS) or a chronic inflammatory pathology, especially intestinal, and the recent occurrence of an episode of sepsis, the presence of many catheters. According to one embodiment, the subject of the invention is suffering from one of these pathologies.

Acute inflammation is also a major event for radiation victims who would not die immediately. It also relates to acute respiratory distress syndromes by exposure to suffocating, vesicant and irritant gases, and other disorders of organs by acute inflammation. Acute inflammation is also a secondary event to exposure to electromagnetic wave weapons.

Acute inflammation also refers to allergic shock states or severe forms of allergies such as "Quincke's edema".

Acute inflammation is also the consequence of a late revascularization of acute ischemia regardless of the territory (cerebral, cardiac, lower limbs), crash syndrome, revascularization after cardiac arrest, severe acute pancreatitis or no, a neuro-malaria (pernicious access), acute thrust of lymphoma or vasculitis.

Acute inflammation also affects the consequences of immune failure in nosocomial infections with more and more opportunistic pathogens. According to another embodiment, the subject of the invention is suffering from severe sepsis. Severe sepsis within the meaning of the present invention is sepsis with acute dysfunction of one or more organs ("multi-visceral failure").

According to another embodiment, the subject of the invention is suffering from septic shock. According to one embodiment, the subject of the invention is an apparently healthy subject, which means that the subject has not been previously diagnosed or identified as having or suffering from a septic state, or who is in the process of develop a septic state or SIRS of similar severity.

According to one embodiment, the subject may be an asymptomatic subject of a septic state. As used herein, an "asymptomatic" subject refers to a subject who does not exhibit the classic symptoms of sepsis.

According to another embodiment, the subject may be at risk of having or developing a septic state, as defined by clinical indices such as, for example: ethnic origin, age, comorbidity, abuse alcohol, poverty, low socio-economic status, or the season (sepsis is more common in winter in areas where it exists).

In another embodiment, the subject has at least two of the following symptoms: temperature greater than 38.2 ° C (or hypothermia less than 36 ° C), tachypnoea greater than or equal to 30 movements per minute (min), upper tachycardia at 120 beats per min, systolic blood pressure less than HO mmHg.

In another embodiment, the subject has at least two of the following symptoms: temperature greater than 38.2 ° C (or hypothermia less than 36 ° C), tachypnoea greater than or equal to 22 movements per minute (min), confusional syndrome ; tachycardia greater than 120 beats per min, systolic blood pressure less than 100 mmHg. According to one embodiment, the subject is an infant or a child with factors promoting the development of septic states. Factors promoting the development of septic status in infants include but are not limited to: immunocompetence (purpura fulminans), immunodeficiency (congenital or acquired immune deficiency), comorbidity (cardiac or urinary malformations), burns, polytrauma, or hospitalized in intensive care.

According to another embodiment, the subject is a child with factors promoting the development of sepsis. Examples of infections that cause sepsis include but are not limited to: meningitis, sepsis, focal infections, respiratory infections, primary bacteremia, genitourinary, abdominal, soft tissue infections , central nervous system, endocarditis. Examples of focal infections include but are not limited to: pneumonia, otitis, epiglottis, conjunctivitis, arthritis, urethritis, pericarditis.

According to another embodiment, the subject of the invention has factors promoting a rapid evolution towards an acute inflammatory reaction and which include but are not limited to: a) sepsis: sepsis, pulmonary infection (such as pneumonia) , intra-abdominal infection, severe purpura infection, necrotic-bullous lesions of necrotizing fasciitis, nosocomial infections, peritonitis, meningitis or bacterial sepsis, b) SIRS, pancreatitis, extensive burn, asthma severe acute, polytrauma, massive irradiation, severe acute asthma, fire fumes, irritating (fighting) gases, ischemia reperfusions as in crush syndrome and molecular excitations of non-lethal weapons.

According to another embodiment, the subject of the invention has undergone a heavy surgical procedure, a surgical procedure with clamping (ischemia-reperfusion), a surgical procedure with extra-corporal circulation.

Thus, the subject of the invention is a method according to the invention, in which the subject is a patient suffering from sepsis and / or generalized acute inflammation, and / or early sepsis, and / or sepsis. serious, and / or septic shock, and / or failures multi-visceral, and / or has at least two of the following symptoms: temperature greater than 38.2 ° C (or hypothermia less than 36 ° C), tachypnoea greater than or equal to 30 movements per minute (min), preferably greater than or equal to 22 movements per minute (min), (tachycardia greater than 120 beats per min), systolic blood pressure less than 100 mm Hg, confusional syndrome; and / or is a subject, particularly an infant or child with factors promoting the development of sepsis or sepsis or factors promoting rapid progression to an acute inflammatory reaction underwent major surgery, clamped-out surgery (ischemia-reperfusion) or extracorporeal circulation.

In a fifth aspect, the invention relates to a kit as described above, for its use in the treatment of cancers. According to one embodiment, the invention relates to a kit as described above, for its use in the treatment of leukemias, preferably acute forms of leukemia, said use implementing the method of administration according to the invention described above. Preferably, the method and the kit according to the invention are used as an adjunct treatment for the treatment of leukemias, preferably acute forms of leukemia. In particular, the kit according to the invention is used as an adjunct treatment, especially during surgery, to limit the proliferation of cancer cells whose modes of attachment to the endothelium have similarities with hyperactivated polynuclear cells.

According to one embodiment, the invention relates to a kit as described above, for its use in the treatment of solid tumors.

In a sixth aspect, the invention relates to a kit as described above, for its use for reducing the production of peroxynitrite or other nitro-oxygenated reactive derivatives (eg superoxide anion 0 ₂ -, oxygen singlet 0 ₂ , peroxide of hydrogen H ₂ 0 ₂ , or ozone ₃ ) in a subject who needs it, said use implementing the method of administration according to the invention described above.

In a seventh aspect, the invention relates to a kit as described above for its use in reducing hyper-inflammation in a subject in need thereof. use implementing the method of administration according to the invention described above.

In an eighth aspect, the invention relates to a kit as described above, for its use to reduce the amount of bacteria, or viruses, or parasites or fungal agents in a subject who needs it, said use implementing the method of administration according to the invention described above.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1 is a photograph showing the appearance of the lungs of a dead rat with complete treatment (Salt P + Na ₂ SeO ₃ ). The pulmonary aspect is substantially normal.

Figure 2 is a photograph showing the appearance of the lungs of a rat having received only Salt-P. The pulmonary aspect is little altered.

EXAMPLES

The present invention will be better understood on reading the following examples which illustrate the invention in a nonlimiting manner.

Example 1 Composition According to the Invention

Composition A is a solution of selenium oxidizing compound: sodium selenite of concentration 0.2 mg / ml equivalent Se. Composition A2 is a control solution of water for injection (ppi).

Composition B is a solution of plasma proteins enriched in Salt-P and containing 0.05 mg / ml of Salt-P. In a preferred embodiment, this purified solution comprises heparin binding proteins (HBP) of which ΑΤ III.

Composition B2 is a control solution of 4% albumin diluted at 20 ' Example 2 Inclusion of the Selenium Compound According to the Invention in a Phantom Red Cell

In one embodiment, the oxidized selenium compound according to the invention is included in a ghost red cell, allowing the release of the oxidizing selenium compound at the level of the microcirculation and the specific site of the adhesion of the phagocytic cells or the immune response. innate with endothelial cells. This allows a cytotoxic action limiting hyper-inflammation and limiting the adhesion of phagocytic cells or the innate immune response with endothelial cells, which involves disulfide bonding and directly or indirectly protect the endothelium.

Example 3 Device according to the invention

The invention further comprises a device according to the invention which administers the selenium protein (in this example, selenoprotein P) slowly then the selenium compound (in this example, sodium selenite) in flash. For example, the delivery device may be the Swiss Medical Care System: Computed Tomography (CT) Express III ™ Contrast Media Delivery System (CMDS), or CT Premica ™ CMDS described in patent applications: EP1713528, EP2298382, EP2477677 .

For example, the delivery device may also be the Medrad Inc. system described in International Application WO2005104687 or US2004242996.

Example 4 Kit according to the invention

The kit according to the invention comprises at least two separate containers, one containing the compound A and the other comprising the compound B: Container of the composition A: 0.15 ml of a solution of sodium selenite at the concentration of 0.2 mg / ml equivalent Selenium (Se). Container of Composition B: 1 ml of a solution of plasma enriched in Salt-P at a concentration of 0.05 mg / ml.

Example 5 Effect of Successive Administrations of Sel-P and Na 2 SeC

Material and methods

rats

Animal care and experimental procedures have been approved by the Ethics Committee and validated according to the procedure in force with the Ministry.

Experiments will be conducted with 12-week old IOPS Han Ico Wistar male rats. (Charles River, L'Arbresle, France). The animals have a weight of between 380 and 400 g at the time of the experiment. They will be kept in individual thermoformed polystyrene cages in accordance with the standards of the Ministry of Agriculture and Environment and kept at a temperature of 21 ± 1 ° C, with a relative humidity of 55% and an alternation of 12h. of the diurnal cycle (day / night). They will have free access to a suitable diet and free access to water corresponding to an adequate intake of selenium. An acclimatization of one week will be done to limit the experimental stress. Animals that are obviously sick at the beginning of the experiments will be excluded.

Groups of animals:

Group 1: Sick Witnesses LPS alone: no intervention.

Group 2: 30 min infusion controls: Administration of control solution B2 over 30 minutes (min.), Wait 10 min. then administration of Na ₂ SeO ₃ (A) over 30 min (infusion).

Group 3: Controls of compositions B and A: administration of control solution B2 over 30 min, wait 10 min. then administration of the control solution A2 in 3 flash injections every 10 min.

Group 4: Action of composition A alone: administration of control solution B2 over 30 min, wait 10 min, then administration of Na ₂ SeO ₃ (A) in 3 flash injections every 10 min. • Group 5: Action of composition B alone: administration of Sel-P (B) over 30 min, wait 10 min. then administration of the control solution A2 in 3 flash injections every 10 min.

• Group 6: Synergistic action of compositions B and A: administration of Sel-P (B) over 30 min, wait 10 min. then administration of Na ₂ SeO ₃ (A) in 3 flash injections every 10 min.

The main criterion is the plasma lactate concentration as reported to the Ethics Committee.

solutions

· LPS: LPS Salmonella typhimurium Serotype LG511, Sigma L-6511 (Lot

12K4090).

• Composition A (sodium selenite): The working solution is at a concentration of 0.2 mg / ml equivalent Se. Injection at H 3h40 after injection of LPS into the internal jugular vein by the central catheter of the amount of the working solution to be made between two lines (corresponding to 0.2 mg / ml) so as to deliver 0.1 mg / kg of selenium equivalent in the form of sodium selenite. Perfusion maintained with physiological saline at least 0.1 ml / h (NaCl).

• Composition A2: Water for injection (ppi) which will be to inject according to the same modalities and rinsing according to the same modalities.

Composition B: the solution of human plasma enriched with selenoprotein-P at the rate of 0.05 mg / ml, with a purification of approximately 8% with a protein concentration of 2 g / l. Injection of 2.5 ml in 30 minutes to each animal.

· Composition B2: 4% human albumin to be diluted at 20 ^th - Injection of 1 ml of the solution diluted to 20 ^th to the animals over 30 min at the rate of 2 ml / h.

Experimental protocol :

H0: Seizure of septic shock by intraperitoneal administration of LPS at 50 mg / kg at H0. Four control rats initially had a dose of 50 mg / kg LPS. Hl, the rats are anesthetized and paired. The anesthesia is performed under penthotal and fentanyl, the animals are then tracheotomized and a catheter is placed in the jugular. The infusion is maintained with physiological saline at a minimum of 0.1 ml / hr (NaCl). - Intervention at H3: Continuous infusion of Salt-P (B) (1 ml) or control B2 (1 ml) over 30 min. Wait for 10 minutes. Continuous infusion is maintained to limit losses of composition B (or B2).

Intervention at H 3h40, 3 ultra rapid injections or flash in 0.5 sec of 0.15 ml of composition A (sodium selenite) or composition A2 (control) every 10 min. and rinsing the infusion with 0.15 ml saline. The injection is performed directly at the internal jugular catheter.

General anesthesia is maintained by administering a half-dose of penthotal intraperitoneally every hour or if the onset of waking sign begins.

Sampling at 6:30 am: Sampling and euthanasia: terminal sampling and euthanasia at 6:30 am after LPS injection, or immediate sampling if the animal dies during the experimental protocol.

Collection of a blood gas and removal of a tube of EDTA and lithium heparin during the euthanasia of the rats. A lung sample was taken weighed and put in the freezer so that at least the dry / wet ratio could be determined as an indicator of pulmonary edema.

Measured parameters:

Immediate determination of lactate, venous blood gases, blood ionogram, blood glucose and hemoglobin levels and serum creatinine hematocrit. Further measurements will be made on samples put in freezing on EDTA and lithium heparin tubes. Results

Effect of treatment on lactate concentration

Control groups include: groups 1; 2 and 4 with sodium selenite in continuous administration or flash administration, group 3 with albumin injection and water injection and group 5 with administration of Sel-P.

Table 2.

Animals from group 4 do not survive the administration of sodium selenite alone. Therefore no value is indicated in Table 2.

In group 6 as a whole, lactate concentration was significantly lowered in rats in this group compared to control rats. The lactate concentration is 2.28 ± 0.31 vs 6.48 ± 1.88 mmol / L (p = 0.017 Mann Whitney test) (Table 2).

This is related to the normalized value of lactate concentration and the very reliable variation between lactate concentrations in this group in a very severe LPS model (50 mg / kg LPS) and euthanasia at 6:30 after administration. of LPS. In addition, the three rats in group 6 (B: selenoprotein-P over 30 minutes and 3 flash injections of A: sodium selenite not hydrated at a dose of 0.1 mg / kg by flash injection) have biological constants close to the values despite administration 6 hours before a lethal dose of LPS at 50 mg / kg. Effect of treatment on blood gases:

Blood gases were measured in venous in euthanized animals (Table 3).

In group 6: the central venous mixed blood saturation is high which indicates the absence or low metabolic suffering, as well as the adapted cardiac output. In addition, the blood glucose is in the reference values, and the pH slightly modified. At the renal level, there is little elevation of serum creatinine concentration.

There is also a macroscopic appearance in the lung which is practically normal and very different from the macroscopic appearance of the control rats, which in this very severe model have a hepatised appearance which is a witness of a very important acute respiratory distress syndrome (Figure 1). In group 2: the table shows that a rat which has received diluted albumin and sodium selenite at a dose of 3 x 0.1 mg / kg = 0.3 mg / kg of selenium over 30 minutes exhibits very different biological parameters compared to group 1 of control rats with very significant pre-lethal suffering: an increase in the major lactate, a lowered pH, a collapsed glucose and an increase in creatinine are observed. The Scv0 ₂ collapsed from probably to very low cardiac output imposing cells maximum oxygen extraction despite the situation of sepsis.

In group 5: in rats that received only selenoprotein-P, there is an incomplete improvement. The pH values are lowered and the renal function is slightly altered as well as the blood glucose value compared to that observed in the previous rat (group 2), corresponding to that observed in most control rats. Scv0 ₂ also remains in values less altered than in the previous rat. There will also be less deterioration of the pulmonary appearance that does not have the hepatic appearance observed in the control rats (Figure 2).

Table 3.

Claims

Kit comprising a first container comprising a composition comprising at least one selenium antioxidant protein and at least one second container comprising at least one composition comprising at least one oxidizing selenium compound.

A kit according to claim 1, wherein said selenium antioxidant protein is selected from the group consisting of: selenoprotein P, glutathione peroxidase, thioredoxin reductase, iodothyronine deiodinase, formate dehydrogenase, methionine sulfoxide reductase, selenophosphase synthetase, selenoprotein Pa (SelPa), selenoprotein W (SelW), selenoprotein T (SelT), selenoprotein R (SelR or SelX), selenoprotein of 15 kDa (Sell5), selenoprotein N (SelN), selenoprotein T ( SelToprotein M (SelM), selenoprotein G-rich (G-rich), selenoprotein W2 (SelW2), selenoprotein BthD (BthD), selenoprotein H (SelH), selenoprotein I (Sell), selenoprotein K (SelK), selenoprotein O (SelO), selenoprotein R (SelR), selenoprotein S (SelS) and selenoprotein Pb (SelPb).

Kit according to claim 1 or 2, wherein the composition of the first container comprises a selenium protein in combination with any pharmaceutically acceptable excipient and further comprises proteins of the group of heparin-binding proteins (HBP), preferably antithrombin III (AT III).

A kit according to any one of claims 1 to 3, wherein said oxidized selenium compound is selected from the group consisting of: sodium selenite, selenocysteine, selenodiglutathione, selenomethyl selenocysteine, dimethyl selinoxide, selenocystamine, or chemical synthesis derivatives containing one or more several selenium atoms, a hybrid selenium, fluorinated selenium salt, chlorinated selenium salt, brominated selenium salt, iodinated selenium salt, a selenoxide, a selenium sulphide salt, a telluric selenium salt, a potassium selenium salt, selenium salt of germanium, selenium salt of barium, selenium salt of lead, selenium salt of zinc or a salt of selenium, nitrogene, atonimy selenide, arsenic selenide, bismuth selenide, cadmium selenide, cobalt selenide, selenide of mercury, selenium oxychloride, selenyl chloride, selenium sulfide, selenium disulfide, selenide silver, selenide of indium, selenide of strontium, selenium acid, selenium dioxide, selenium, selenious acid, selenic acid, selenoglutathione.

5. Kit according to any one of claims 1 to 4, wherein said oxidizing selenium compound is sodium selenite, preferably in a non-pentahydrate form, or selenocysteine.

6. Kit according to any one of claims 1 to 5, wherein the composition of the second container comprises a selenious oxidizing compound, in combination with any pharmaceutically acceptable excipient.

The kit according to any one of claims 1 to 6, wherein said oxidant selective compound is included in a transport means selected from the group consisting of: a ghost red blood cell, a capsule, a nanoparticle, a liposome, preferably said transport means is a ghost red blood cell.

8. Kit according to any one of claims 1 to 7, further comprising a third anti-oxidant container comprising vitamin C at low concentration, vitamin E, a glutathione precursor, low concentration copper salts and or zinc salts.

9. Kit according to any one of claims 1 to 8, further comprising a fourth oxidizing container comprising vitamin C in high concentration, iron salts, gold salts, copper salts in high concentration, arsenic salts and / or oxidizing telluric compounds.

Kit according to any one of claims 1 to 9, comprising:

at. a first container, comprising a composition comprising a selenium antioxidant protein, preferably selenoprotein P, b. one, two or at least three second containers, each of the second containers comprising at least one composition comprising at least one oxidizing selenium compound, preferably sodium selenite,

vs. optionally a third antioxidant container comprising low concentration vitamin C, vitamin E, a glutathione precursor, low concentration copper salts and / or zinc salts. d. optionally a fourth oxidizing container comprising vitamin C at high concentration, iron salts, gold salts, high concentration copper salts, arsenic salts and / or tellurium compounds. 11. Kit according to any one of claims 1 to 10 for its use in the treatment of sepsis and / or acute generalized inflammation.

12. Kit according to any one of claims 1 to 10 for its use in the treatment of solid cancers or leukemias, preferably acute forms of leukemia. 13. A method for administering, to a subject in need thereof, a therapeutically effective amount of the compositions included in the different containers of the kit according to any one of claims 1 to 12, in a sequential mode, namely first. the administration of an effective amount of the composition of the first container, which is a composition comprising an antioxidant protein, preferably a selenium protein, continuously, then the administration in a flash or multiple repeated flashes of an effective amount of the composition of the second container, which is a composition comprising an oxidizing selenium compound.

14. The method for administration according to claim 13, comprises the continuous administration over 10 minutes to 1 hour, of an effective amount of selenie protein composition of the first container, which is preferably selenoprotein P, and a few minutes. after an effective amount of the selenium compound composition of the second container or containers, in three repeated flashes.

The method of any one of claims 13 or 14, wherein the selenium protein of the invention is to be or is administered by injection to about 2 to 90 mg of P-selenoprotein continuously for about 10 minutes to about one hour, and 10 to 20 minutes after the end of this administration, the oxidative selenium compound is or is to be administered by injection 0.05 to about 0.6 mg equivalent Se / kg body weight, of compound selenium, preferably sodium selenite, the dose being divided into 3 successive flash injections spaced 10 minutes.

The method according to any one of claims 13 to 15, wherein the subject is suffering from sepsis and / or generalized acute inflammation, and / or early sepsis, and / or severe sepsis, and / or septic shock, and / or multi-visceral failures, and / or has at least two of the following symptoms: temperature greater than 38.2 ° C (or hypothermia less than 36 ° C), tachypnoea greater than or equal to at 22 movements per minute (min), tachycardia greater than 120 beats per min, systolic blood pressure less than

100 mmHg.

The method according to any one of claims 13 to 16, wherein the subject is suffering from solid cancer or leukemia, preferably acute forms of leukemia, and the method according to the invention used in adjunctive treatment for the treatment of leukemias, preferably acute forms of leukemia.

18. A method according to any one of claims 13 to 17 for reducing the production of peroxynitrite, or hyper inflammation in a subject who needs it. 19. Method according to any one of claims 13 to 16, wherein the subject is suffering from acute inflammation due in particular to exposure to irradiation or to an electromagnetic wave, or is suffering from an acute respiratory distress syndrome. , especially due to exposure to suffocating, vesicant or irritant gases.

20. A method according to any one of claims 13 to 16, wherein the subject is suffering from acute inflammation due to allergic shock or allergy.

21. The method of any one of claims 13 to 16, wherein the subject has undergone cardiac arrest or ischemia reperfusion.

22. Method according to any one of claims 13 to 16, wherein the subject is under antibiotic treatment or has a bacterial infection particularly resistant to antibiotics or is suffering from a viral infection.

The method of any one of claims 13 to 16, wherein the subject is an organ and the method is implemented prior to organ harvesting, as part of a graft preparation, as part of a transplant. and / or in the context of transplant rejection.

24. Apparatus for implementing the method according to any one of claims 13 to 23.