Kit for treating sepsis and/or any systemic (SIRS) or damaging cellular hyperinflammation

Abstract

Disclosed is a kit including a first container including a composition including at least one antioxidant selenoprotein and at least one second container including at least one composition including at least one oxidant selenocompound. Also disclosed is a method of administration that allows to administer effective and cytotoxic doses of selenocompounds, allowing the inhibition of the hyper-activation of phagocytes and in particular of circulating immature neutrophils and directly and indirectly protects endothelial cells, in particular for the treatment of sepsis, SIRS and leukemia. Further disclosed is an administration device adapted to the administration method.

Claims

1. Method for treating sepsis and/or acute systemic inflammation, solid cancers or leukemia in a subject in need thereof, said method comprising sequentially administering to the subject: first a therapeutically effective amount of a composition comprising at least one antioxidant selenoprotein by continuous administration over 10 minutes to 1 hour, wherein the therapeutically effective amount is 2 to 90 mg of the selenoprotein; and then a therapeutically effective amount of at least another composition comprising at least one oxidant selenocompound by administration in one flash in a time of 0.1 to 4 seconds or in several repeated flashes in a time of 0.1 to 4 seconds each, wherein the therapeutically effective amount per each administered flash is 0.05 to 0.6 mg of Se equivalent/kg of body weight of the selenocompound.

2. Method according to claim 1, wherein the therapeutically effective amount of the at least another composition comprising at least one oxidant selenocompound is administered to the subject in three repeated flashes.

3. Method according to claim 1, wherein the therapeutically effective amount of the composition comprising at least one antioxidant selenoprotein is administered to the subject by continuous injection of 2 to 90 mg of selenoprotein P in its complete form or in truncated forms over 10 minutes to 1 hour, and wherein the therapeutically effective amount of the at least another composition comprising at least one oxidant selenocompound is administered to the subject 10 to 20 minutes after the end of the first administration by injection of 0.05 to 0.6 mg of Se equivalent/kg of body weight of the selenocompound, the dose being fractioned into 3 successive flash injections, in a time of 0.1 to 4 seconds each, at intervals of 10 minutes.

4. Method according to claim 1, wherein the subject is suffering from sepsis and/or acute systemic inflammation and/or sepsis in the early stage, and/or a severe sepsis, and/or a septic shock and/or multi-organ failures, and/or has at least two of the following symptoms: temperature higher than 38.2° C., hypothermia lower than 36° C., tachypnea higher than or equal to 22 movements per minute, tachycardia higher than 120 beats per minute, or systolic arterial pressure lower than 100 mmHg.

5. Method according to claim 1, wherein the subject is suffering from a solid cancer or leukemia.

6. Method according to claim 1, wherein said method allows to reduce the production of peroxynitrite or to reduce hyper inflammation in the subject in need thereof.

7. Method according to claim 1, wherein the subject is suffering from an acute inflammation or is suffering from an acute respiratory distress syndrome.

8. Method according to claim 1, wherein the subject is suffering from an acute inflammation due to an allergic shock or an allergy; or has experienced a cardiac arrest or an ischemia reperfusion; or is undergoing antibiotic treatment or is suffering from a bacterial infection or a viral infection.

9. Method according to claim 1, wherein said composition comprising at least one antioxidant selenoprotein further comprises a pharmaceutically acceptable excipient and proteins from the group of heparin-binding proteins (HBP).

10. Method according to claim 1, wherein said antioxidant selenoprotein is selected from the group consisting of selenoprotein P, glutathione peroxidase, thioredoxin reductase, iodothyronine deiodinase, formate dehydrogenase, methionine sulfoxide reductase, selenophosphate synthetase, selenoprotein Pa (SelPa), selenoprotein W (SelW), selenoprotein T (SelT), selenoprotein R or selenoprotein X (SelR or SelX), 15 kDa selenoprotein (Sel15), selenoprotein N (SelN), selenoprotein T2 (SelT2), selenoprotein M (SelM), G-rich selenoprotein (G-rich), selenoprotein W2 (SelW2), selenoprotein BthD (BthD), selenoprotein H selenoprotein I (SelI), selenoprotein K (SelK), selenoprotein O (SelO), selenoprotein S (SelS), and selenoprotein Pb (SelPb).

11. Method according to claim 1, wherein said antioxidant selenoprotein is selenoprotein P in its complete form or in truncated forms.

12. Method according to claim 1, wherein said oxidant selenocompound is selected from the group consisting of sodium selenite, selenocysteine, selenodiglutathione, selenomethyl selenocysteine, dimethyl selinoxide, selenocystamine, derivatives of chemical synthesis containing one or several selenium atoms, a hybrid selenium, selenium fluoride salt, selenium chloride salt, selenium bromide salt, selenium iodide salt, a selenoxide, a selenium sulfide salt, selenium telluride salt, selenium potassium salt, selenium germanium salt, selenium barium salt, selenium lead salt, selenium zinc salt or a nitrogenated selenium salt, antimony selenide, arsenic selenide, bismuth selenide, cadmium selenide, cobalt selenide, mercury selenide, selenium oxychloride, selenyl chloride, selenium sulfide, selenium disulfide, silver selenide, indium selenide, strontium selenide, selenic acid, selenium dioxide, selenium, selenous acid, and selenoglutathione.

13. Method according to claim 1, wherein said oxidant selenocompound is sodium selenite or selenocysteine.

14. Method according to claim 1, wherein said oxidant selenocompound is included in a transport means selected from the group consisting of a ghost red blood, a capsule, a nanoparticle, and a liposome.

15. Method according to claim 1, wherein the administration steps are repeated every 4, 6, 8 or 12 hours for at least one day.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is a photograph showing the appearance of the lungs of a deceased rat with the complete treatment (Sel P+Na.sub.2SeO.sub.3). The appearance of the lungs is sensibly normal.

(2) FIG. 2 is a photograph showing the appearance of the lungs of a rat having received only Sel-P. The appearance of the lungs shows little alteration.

EXAMPLES

(3) This invention will be better understood after reading the following examples that illustrate the invention in a non-limitative manner.

Example 1: Composition According to the Invention

(4) Composition A is a solution of oxidant selenocompound: sodium selenite with a concentration of 0.2 mg/ml in Se equivalent.

(5) Composition A2 is a control solution of water for injection (wfi).

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

(7) Composition B2 is control solution of 4% albumin diluted to 1:20.

Example 2: Inclusion of the Selenocompound According to the Invention in a Ghost Red Blood Cell

(8) In one embodiment, the oxidant selenocompound according to the invention is included in a ghost red blood cell, allowing the liberation of the oxidant selenocompound in the microcirculation and at the specific site of the adhesion of phagocyte cells or cells of the innate immune response with the endothelial cells. This enables a cytotoxic action limiting hyper-inflammation and limiting adhesion of the phagocyte cells or the cells of the innate immune response with the endothelial cells, adhesion that involves disulfide bridges and to directly or indirectly protect the endothelium.

Example 3: Device According to the Invention

(9) The invention also comprises a device according to the invention that administers the selenoprotein (in this example, selenoprotein P) slowly and then the selenocompound (in this example, sodium selenite) in flash.

(10) For example, the administration 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.

(11) For example, the administration device may also be the Medrad Inc. system described in international application: WO2005104687, or application US2004242996.

Example 4: Kit According to the Invention

(12) The kit according to the invention comprises at least two distinct containers, one containing compound A and the other containing compound B:

(13) Container of composition A: 0.15 ml of a solution of sodium selenite with a concentration of 0.2 mg/ml in Selenium (Se) equivalent.

(14) Container of composition B: 1 mL of a plasma solution enriched in Sel-P at a concentration of 0.05 mg/mL.

Example 5: Effect of Successive Administrations of Sel-P and Na.SUB.2.SeO.SUB.3

(15) Material and Methods

(16) Rats

(17) Care of the animals and experimental procedures were approved by the ethics committee and validated following the procedure in force with the Ministry.

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

(19) Animal Groups:

(20) Group 1: Diseased controls LPS only: no action. Group 2: 30-minute perfusion controls: administration of control solution B2 over 30 minutes (min.), 10 min. wait then administration of Na.sub.2SeO.sub.3 (A) over 30 min (perfusion). Group 3: Compositions B and A controls: administration of control solution B2 over 30 min, 10 min. wait then administration of 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, 10 min wait, then administration of Na.sub.2SeO.sub.3 (A) in 3 flash injections every 10 min. Group 5: Action of composition B alone: administration of Sel-P (B) over 30 min, 10 min. wait then administration of control solution A2 in 3 flash injections every 10 min. Group 6: Action of compositions B and A in synergy: administration of Sel-P (B) over 30 min, 10 min. wait then administration of Na.sub.2SeO.sub.3 (A) in 3 flash injections every 10 min.

(21) The main criterion is the plasma concentration of lactate as was mentioned to the ethics committee.

(22) Solutions

(23) LPS: LPS Salmonella typhimurium Serotype LG511, Sigma L-6511 (Batch 12K4090). Composition A (sodium selenite): The working solution is at a concentration of 0.2 mg/ml of Se equivalent. Injection at H 3 h40 after the injection of LPS into the internal jugular vein through the central catheter of the amount of 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 by physiological serum, minimum 0.1 ml/h (NaCl). Composition A2: Water for injection (wfi) that will be to inject according to the same procedure and rinsing according to the same procedure. Composition B: the solution of human plasma enriched with selenoprotein-P at 0.05 mg/ml, with a purification of about 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 to 1:20—Injection of 1 ml of the solution diluted to 1:20 to the animals over 30 min at a rate of 2 ml/h.
Experimental Protocol: H0: Triggering of the septic shock by the intra-peritoneal administration of LPS at 50 mg/kg at H0. 4 control rats were initially given a dose of 50 mg/kg of LPS. H1, the rats are anaesthetized and instrumented. The anesthetic is applied in the form of penthotal and fentanyl, the animals are then tracheostomized and a catheter is put into place in the jugular. Perfusion is maintained by physiological serum, with a minimum of 0.1 ml/h (NaCl). Operation at H3: Continuous perfusion of Sel-P (B) (1 ml) or control B2 (1 ml) over 30 min. Wait of 10 minutes. Continuous perfusion is maintained to limit losses of composition B (or B2). Operation at H 3 h40, 3 ultra-fast or flash injections in 0.5 sec of 0.15 ml of composition A (sodium selenite) or composition A2 (control) every 10 min. and rinsing of the perfusion with 0.15 ml of physiological serum. The injection is made directly at the internal jugular catheter.

(24) General anesthesia is maintained by intraperitoneal administration of a half-dose of penthotal every hour or at the first sign of awakening.

(25) Samples Taken at H 6 h30:

(26) Sampling and euthanasia: final sampling and euthanasia at 6 h30 after the injection of LPS, or immediate sampling if the animal dies during the experimental protocol.

(27) Collection of a blood gas sample and collection of a tube of EDTA and lithium heparinate at the time of euthanasia of the rats.

(28) A lung sample was taken, weighed and placed in the freezer in order to be able to determine, at least, the dry/wet ratio as a pulmonary edema indicator.

(29) Measured Parameters:

(30) Immediate measurement of lactate, of gases in venous blood, of the blood ionogram, of glycemia and of the hemoglobin level and of the hematocrit of the creatinine level. Other measurements will be made on the samples frozen on EDTA and lithium heparinate tubes.

(31) Results

(32) Effect of the Treatment on the Lactate Concentration:

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

(34) TABLE-US-00002 TABLE 2 Group: Lactate level Controls 1 4.78 1 3.76 1 5.45 1 7.36 2 8.75 3 6.78 3 8.45 Treatment: 5 3.35 Sel-P 5 4.79 Treatment in 6 2.2 synergy 6 2.63 6 2.02

(35) Animals in group 4 do not survive the administration of sodium selenite alone. Consequently, no value is shown in Table 2.

(36) For the group 6 overall, the lactate concentration is significantly reduced in rats of this group compared to the rats of the control groups. The lactate concentration is 2.28±0.31 vs 6.48±1.88 mmol/L (p=0.017 Mann Whitney test) (Table 2).

(37) This is related to the normalized value of the lactate concentration and to the very small variation between lactate concentrations in this group in a model of very severe LPS (50 mg/kg of LPS) and euthanasia at 6 h30 after the administration of LPS.

(38) Furthermore, the three rats of group 6 (B: selenoprotein-P over 30 minutes and 3 flash injections of A: non-hydrated sodium selenite at a dose of 0.1 mg/kg by flash injection) have biological constants close to the reference values despite the administration 6 h30 earlier of a fatal dose of LPS at 50 mg/kg.

(39) Effect of the Treatment on Blood Gases:

(40) Venous blood gases have been measured in euthanized animals (Table 3).

(41) In group 6: saturation in mixed central venous blood is high which is a sign of the absence or low level of metabolic distress, and of the adapted heart output. Furthermore, glycemia is within the reference values, and the pH is slightly modified. At the kidney level, there is very little increase in the concentration of creatinine.

(42) Furthermore, the macroscopic appearance of the lungs is almost normal, which is very different from the macroscopic appearance in control rats which, in this very severe model, have a hepatized aspect, a sign of a very severe acute respiratory distress syndrome (FIG. 1).

(43) In group 2: the table shows that a rat having received diluted albumin and sodium selenite at a dose of 3×0.1 mg/kg=0.3 mg/kg of selenium over 30 minutes presents very different biological parameters from the group 1 of control rats with very severe, pre-lethal distress: a major increase in lactate is observed, with a lowered pH, a very low glycemia and an increase in creatinine. ScvO.sub.2 collapsed probably due to very low heart output imposing to cells a maximum oxygen extraction despite the sepsis situation.

(44) In group 5: an incomplete improvement is observed in rats having only received selenoprotein-P. pH values are lowered and the kidney function is little changed, as is the value of glycemia compared with what is observed in the previous rat (group 2), corresponding to what is observed in most control rats. ScvO.sub.2 also remains within values less altered than in the previous rat. There is also a smaller change to the lung appearance which does not have the hepatized aspect observed in control rats (FIG. 2).

(45) TABLE-US-00003 TABLE 3 Group: 1 1 1 1 2 3 3 5 5 6 6 6 pH 7.44 7.270 7.43 6.74 6.906 7.240 7.210 7.357 7.346 7.400 7.491 7.313 PaO.sub.2 387 33.7 40.9 59.6 16 37.9 33.7 55.3 34 51.5 40.4 47.8 mmHg PCO.sub.2 20.1 83.4 33.2 44.9 97 39.9 56.5 44.7 45.4 48.7 34.1 54.3 mmHg CO.sub.3H— 13.7 37.8 23.3 6.1 19.6 18.6 22.2 25.1 27.7 31 26.6 28.3 ScvO.sub.2 or 100 54 79.3 58.3 8.9 65 51.2 86.9 64.5 85.9 80.7 79.3 SaO.sub.2 Probable 4.78 5.45 7.36 arterial lactate Venous 3.76 8.75 6.78 8.45 3.35 4.79 2.2 2.63 2.02 lactate Na 139 143 141 155 144 105 132 140 143 145 144 142 K 4.5 5.8 4.2 5 7.5 >12 9.9 4.6 5.2 6.4 5.2 6.1 Cl 116 104 105 133 116 81 103 107 105 107 108 106 Ca 1.01 1.16 1.08 0.62 1.21 0.57 0.71 1.12 1.17 1.12 1.14 1.22 AgapK 14 7 17 21 16 ND 17 13 16 13 15 14 Ht 40 48 38 <10% 35 45 50 38 41 35 38 35 cHb 13.5 16.4 12.9 NA 12.1 15.4 17 13 14 11.8 13 11.9 Gly 1.1 1.01 1.2 1.21 0.27 0.52 0.67 1.16 1.25 1.14 1.23 1 Creat 77 180 113 44 156 132 215 52 94 100 78 113