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Toxicity and Pharmacokinetics of novel IL-1 Trap heterodimeric fusion protein
Abstracts / Toxicology Letters 238S (2015) S56–S383
tective effects of granulocyte colony stimulating factor (G-CSF) on cardiomyocyte apoptosis after CO poisoning in rats. Methods: Male Wistar albino rats were exposed to CO 1500 or 3000 ppm for 60 min. Single and multiple doses of G-CSF (10, 50, and 100 g/kg) were administered to 17 groups (N = 5) following CO 1500 ppm exposure and 17 groups (N = 5) following CO 3000 ppm exposure. After CO poisoning, carboxyheamoglobin concentration were measured for all groups. Also, cardiomyocyte apoptosis in all groups were assessed via determination of caspase 3 activity by immunofluorescence and for the group poisoned with CO 3000 ppm and treated with G-CSF 100 g/kg apoptosis was determined byterminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Results: Concentration of carboxyheamoglobin in blood significantly increased following exposure to CO 1500 ppm (19–47%) and 3000 ppm (60–76%). However, carboxyheamoglobin levels were significantly higher in CO 3000 ppm group as compared to CO 1500 ppm (p < 0.05). Caspase 3 activity differences between G-CSF and control groups were significant and G-CSF could decrease apoptosis following CO 3000 ppm poisoning (p < 0.001). The greatest decrease in caspase 3 activity was seen in animals exposed to CO 3000 ppm and treated with multiple doses of G-CSF 100 g/kg. TUNEL assay showed that in G-CSF 100 g/kg multiple doses treated group, apoptosis was significantly ameliorated in comparison to control and sham groups (p < 0.05). Conclusions: In this study, we confirmed that CO poisoning leads to cardiomyocytes apoptosis which could be significantly reduced by G-CSF treatment.
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administration at 200 mg/kg. Serum was prepared and analyzed by immunoassay. The following evaluations were performed during the study: clinical signs, body weights, clinical pathology, and pharmacokinetic analysis. Results: There were no test item-related deaths, and no clinical signs of local or systemic toxicity observed during the study. The No Observed Adverse Effect Level was considered to be 200 mg/kg for both the sc and iv routes of administration. Following sc and iv administrations, elimination of RPH-104 was complete by 504 h post-dose. The mean half-life of RPH-104 was 28–87 h and 52–70 h for sc and for iv administration respectively, with no trend relating to dose level or animal gender. Following both sc and iv administrations of RPH-104, increases in maximum concentrations and mean exposures to RPH-104 were essentially proportional with respect to dose level, indicating that there was no saturation of routes of absorption or elimination with increasing dose level. http://dx.doi.org/10.1016/j.toxlet.2015.08.889
P13-111 Dissolution of alkaline earth silicate wool and polycrystalline wool in saline solution A. Campopiano 1,∗ , A. Cannizzaro 1 , F. Angelosanto 1 , E. Barrese 2 , S. Iavicoli 1 1 2
INAIL, Monteporzio Catone (Rome), Italy INAIL, Lamezia Terme (CZ), Italy
http://dx.doi.org/10.1016/j.toxlet.2015.08.888
P13-110 Toxicity and Pharmacokinetics of novel IL-1 Trap heterodimeric fusion protein A. Krotkova 1,∗ , E. Shipaeva 1,∗ , C.M. Luetjens 2 , F.T. Ludwig 2 , T. Hardwick 3 , Y. Lavrovsky 4 , M. Samsonov 1 1
CJSC «R-Pharm», Mocow, Russian Federation Covance Preclinical Services GmbH, Münster, Germany 3 Covance Laboratories Ltd., Harrogate, United Kingdom 4 R-Pharm Overseas Inc., San Diego, United States 2
Signal transduction through the IL-1 receptor type I (IL-1RI) pathway has been shown to mediate a strong pro-inflammatory response, which contributes to a number of human autoinflammatory diseases such as diabetes, cryopyrin-associated periodic syndromes, Behc¸et Disease, Familial Mediterranean Fever, gout and certain types of arthritis. RPH-104 is a novel heterodimeric fusion protein capable of binding to human IL-1, IL-1a and IL-1 receptor agonist (IL-1RA). The RPH-104 heterodimer comprised of soluble portions of human extracellular portions of IL-1RI and IL-1 receptor accessory protein (IL-1RAcP) each linked to a mutant Fc portion of human IgG1 fragment. Objectives: This study aimed to assess the toxicity and pharmacokinetics of RPH-104 following a single subcutaneous (sc) or intravenous (iv) (bolus) administration in the cynomolgus monkey, followed by a 30-day recovery period. Methods: 24 naive male and female cynomolgus monkeys were allocated to 6 groups and dosed with either the vehicle, 5, 50 and 200 mg/kg RPH-104 (sc, Groups 1–4) or 5 and 200 mg/kg RPH-104 (iv, Groups 5 and 6) on Day 1. Following dosing, blood samples were collected pre-dose and at 0.5, 1, 4, 8, 24, 48, 96, 168, 336, 504 and 696 h post-dose. Additional blood samples were taken from animals at 0.083 and 0.25 h post-dose following both sc and iv
The possible pathogenicity of a fiber depend upon the extent to which the fibers can be inhaled and can persist in the lung. Biopersistence of fibers is determined by the rate at which they are physically cleared from the lung by mechanical action and the rate at which they dissolve in the lung (biodurability). Biodurability may involve processes occurring both within cells and in the extracellular fluids. Among man made mineral fibers, the alkaline earth silicate wool (AESW) and polycrystalline wool (PCW), new generation wools of high-temperature insulation materials having low biopersistence, are replacing ceramic fibers in some applications. For these materials, IARC decided not to make an overall evaluation because no human data were available, although such fibers appear to have low carcinogenic potential in experimental animals. The aim of this paper is to study the behaviour of AESW and PCW in saline solution simulating physiological fluids. AESW is composed principally of SiO2 (61–67%) and CaO (27–33%), and PCW of Al2 O3 > 72% and SiO2 . To simulate macrophage environment and extracellular environment of the lung, a Gamble’s solution to two different pH (4.5 and 7.4) was used. The stationary test lasted from 1 to 4 weeks. Morphological and physical changes of fibers that accompanied the leaching and dissolution process are studied by scanning electron microscopy (SEM). Each solution was analyzed by inductively coupled plasma atomic emission spectrometry (ICPOES) to quantify the concentration of the elements extracted from wools. The amount of silicon released in the solution at pH 7.4 rose during the four weeks of treatment, much more for AESW than PCW. The SEM analysis showed no variation of length-weighted fiber diameter (DLG ≈ 4 m) for PCW in both environments; and an almost complete leaching of AESW in the pH 7.4 solution. For PCW, the dissolution seemed to produce uncorroded fibers with the same chemical composition as the untreated fibers. The dissolution fiber leads to breakage transverse of fiber and the DLG value is affected little of this change. AESW showed a dissolution highly non-congruent with rapid leaching of calcium. The chemical composition of the fibers and the pH environment have a strong
tective effects of granulocyte colony stimulating factor (G-CSF) on cardiomyocyte apoptosis after CO poisoning in rats. Methods: Male Wistar albino rats were exposed to CO 1500 or 3000 ppm for 60 min. Single and multiple doses of G-CSF (10, 50, and 100 g/kg) were administered to 17 groups (N = 5) following CO 1500 ppm exposure and 17 groups (N = 5) following CO 3000 ppm exposure. After CO poisoning, carboxyheamoglobin concentration were measured for all groups. Also, cardiomyocyte apoptosis in all groups were assessed via determination of caspase 3 activity by immunofluorescence and for the group poisoned with CO 3000 ppm and treated with G-CSF 100 g/kg apoptosis was determined byterminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Results: Concentration of carboxyheamoglobin in blood significantly increased following exposure to CO 1500 ppm (19–47%) and 3000 ppm (60–76%). However, carboxyheamoglobin levels were significantly higher in CO 3000 ppm group as compared to CO 1500 ppm (p < 0.05). Caspase 3 activity differences between G-CSF and control groups were significant and G-CSF could decrease apoptosis following CO 3000 ppm poisoning (p < 0.001). The greatest decrease in caspase 3 activity was seen in animals exposed to CO 3000 ppm and treated with multiple doses of G-CSF 100 g/kg. TUNEL assay showed that in G-CSF 100 g/kg multiple doses treated group, apoptosis was significantly ameliorated in comparison to control and sham groups (p < 0.05). Conclusions: In this study, we confirmed that CO poisoning leads to cardiomyocytes apoptosis which could be significantly reduced by G-CSF treatment.
S311
administration at 200 mg/kg. Serum was prepared and analyzed by immunoassay. The following evaluations were performed during the study: clinical signs, body weights, clinical pathology, and pharmacokinetic analysis. Results: There were no test item-related deaths, and no clinical signs of local or systemic toxicity observed during the study. The No Observed Adverse Effect Level was considered to be 200 mg/kg for both the sc and iv routes of administration. Following sc and iv administrations, elimination of RPH-104 was complete by 504 h post-dose. The mean half-life of RPH-104 was 28–87 h and 52–70 h for sc and for iv administration respectively, with no trend relating to dose level or animal gender. Following both sc and iv administrations of RPH-104, increases in maximum concentrations and mean exposures to RPH-104 were essentially proportional with respect to dose level, indicating that there was no saturation of routes of absorption or elimination with increasing dose level. http://dx.doi.org/10.1016/j.toxlet.2015.08.889
P13-111 Dissolution of alkaline earth silicate wool and polycrystalline wool in saline solution A. Campopiano 1,∗ , A. Cannizzaro 1 , F. Angelosanto 1 , E. Barrese 2 , S. Iavicoli 1 1 2
INAIL, Monteporzio Catone (Rome), Italy INAIL, Lamezia Terme (CZ), Italy
http://dx.doi.org/10.1016/j.toxlet.2015.08.888
P13-110 Toxicity and Pharmacokinetics of novel IL-1 Trap heterodimeric fusion protein A. Krotkova 1,∗ , E. Shipaeva 1,∗ , C.M. Luetjens 2 , F.T. Ludwig 2 , T. Hardwick 3 , Y. Lavrovsky 4 , M. Samsonov 1 1
CJSC «R-Pharm», Mocow, Russian Federation Covance Preclinical Services GmbH, Münster, Germany 3 Covance Laboratories Ltd., Harrogate, United Kingdom 4 R-Pharm Overseas Inc., San Diego, United States 2
Signal transduction through the IL-1 receptor type I (IL-1RI) pathway has been shown to mediate a strong pro-inflammatory response, which contributes to a number of human autoinflammatory diseases such as diabetes, cryopyrin-associated periodic syndromes, Behc¸et Disease, Familial Mediterranean Fever, gout and certain types of arthritis. RPH-104 is a novel heterodimeric fusion protein capable of binding to human IL-1, IL-1a and IL-1 receptor agonist (IL-1RA). The RPH-104 heterodimer comprised of soluble portions of human extracellular portions of IL-1RI and IL-1 receptor accessory protein (IL-1RAcP) each linked to a mutant Fc portion of human IgG1 fragment. Objectives: This study aimed to assess the toxicity and pharmacokinetics of RPH-104 following a single subcutaneous (sc) or intravenous (iv) (bolus) administration in the cynomolgus monkey, followed by a 30-day recovery period. Methods: 24 naive male and female cynomolgus monkeys were allocated to 6 groups and dosed with either the vehicle, 5, 50 and 200 mg/kg RPH-104 (sc, Groups 1–4) or 5 and 200 mg/kg RPH-104 (iv, Groups 5 and 6) on Day 1. Following dosing, blood samples were collected pre-dose and at 0.5, 1, 4, 8, 24, 48, 96, 168, 336, 504 and 696 h post-dose. Additional blood samples were taken from animals at 0.083 and 0.25 h post-dose following both sc and iv
The possible pathogenicity of a fiber depend upon the extent to which the fibers can be inhaled and can persist in the lung. Biopersistence of fibers is determined by the rate at which they are physically cleared from the lung by mechanical action and the rate at which they dissolve in the lung (biodurability). Biodurability may involve processes occurring both within cells and in the extracellular fluids. Among man made mineral fibers, the alkaline earth silicate wool (AESW) and polycrystalline wool (PCW), new generation wools of high-temperature insulation materials having low biopersistence, are replacing ceramic fibers in some applications. For these materials, IARC decided not to make an overall evaluation because no human data were available, although such fibers appear to have low carcinogenic potential in experimental animals. The aim of this paper is to study the behaviour of AESW and PCW in saline solution simulating physiological fluids. AESW is composed principally of SiO2 (61–67%) and CaO (27–33%), and PCW of Al2 O3 > 72% and SiO2 . To simulate macrophage environment and extracellular environment of the lung, a Gamble’s solution to two different pH (4.5 and 7.4) was used. The stationary test lasted from 1 to 4 weeks. Morphological and physical changes of fibers that accompanied the leaching and dissolution process are studied by scanning electron microscopy (SEM). Each solution was analyzed by inductively coupled plasma atomic emission spectrometry (ICPOES) to quantify the concentration of the elements extracted from wools. The amount of silicon released in the solution at pH 7.4 rose during the four weeks of treatment, much more for AESW than PCW. The SEM analysis showed no variation of length-weighted fiber diameter (DLG ≈ 4 m) for PCW in both environments; and an almost complete leaching of AESW in the pH 7.4 solution. For PCW, the dissolution seemed to produce uncorroded fibers with the same chemical composition as the untreated fibers. The dissolution fiber leads to breakage transverse of fiber and the DLG value is affected little of this change. AESW showed a dissolution highly non-congruent with rapid leaching of calcium. The chemical composition of the fibers and the pH environment have a strong