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Valutazione dell’immunotossicità per il nuovo inibitore della tirosin-chinasi della milza R406

Posted on Giugno 15, 2022Giugno 22, 2022 by Andrea
  1. Spleen tyrosine kinase (Syk) is a novel pharmaceutical target in the treatment of allergic, autoimmune and neoplastic diseases. Previous studies have shown that Syk signaling plays a key role in regulating the lymphohematopoietic system. These observations prompted us to investigate whether Syk’s inhibition promoted immunotoxicity.
  2. In a series of studies,  rats  were given orally R406 at doses up to and including 100 mg / kg / day (or its prodrug R788 at doses up to and including 100 mg / kg / day, reduced to 50 mg / day). ). kg / day for women after passing MTD), a strong Syk inhibitor, twice daily for 28 days.
  3. Oltre alle valutazioni tossicologiche standard, sono stati eseguiti test di immunofenotipizzazione citometrica a flusso e di risposta immunitaria umorale misurando i livelli di  anticorpi IgM   e IgG  anti  –  KLH  .  Altri studi di immunotossicità includevano tre modelli di resistenza dell’ospite in topi Balb/c femmine per determinare ulteriormente gli effetti di R406 sull’immunità innata e acquisita.
  4. Dopo il trattamento con R406,  negli studi sui ratti sono stati osservati gli effetti immunomodulatori attesi (ad es. diminuzione del peso del timo e della milza, perdita di cellule del midollo osseo e diminuzione della conta dei linfociti, inclusi i linfociti T e B). Questi cambiamenti si sono generalmente risolti dopo un periodo di recupero di 14 giorni senza trattamento  . KLH   challenge   nei   ratti non   ha mostrato  effetti  avversi sulla risposta IgG o   IgM   . R788/406, somministrato per via orale a dosi fino a 80 mg/kg/die inclusi per 28 giorni, non ha influenzato la clearance batterica o virale nei modelli di resistenza murina rispettivamente di Listeria, streptococco o ospiti dell’influenza.
  5. Ciò era correlato a precedenti test di funzionalità dei macrofagi e dei neutrofili in vitro (valutazione di ioni migratori, fagocitosi, scoppio ossidativo e attività battericida) che hanno mostrato che  l’ R406  non ha influenzato negativamente la funzione dei macrofagi o dei neutrofili nelle risposte immunitarie innate. Presi insieme, questi risultati mostrano che l’  R406  ha un’immunotossicità funzionale minima nonostante i suoi effetti linfocitopenici, suggerendo che l’inibizione di Syk potrebbe non portare a effetti avversi inaccettabili basati sui meccanismi.

Valutazione dell’immunotossicità per il nuovo inibitore della tirosin-chinasi della milza R406.

    • Cdna 8Spleen tirosin chinasi (Syk) è un nuovo target farmaceutico nel  trattamento delle malattie allergiche, autoimmuni e cancerose. Precedenti studi hanno dimostrato che la segnalazione Syk svolge un ruolo chiave nella regolazione del sistema linfoematopoietico.
    • Queste osservazioni ci hanno spinto a indagare se l’inibizione di Syk promuovesse l’immunotossicità. In una serie di studi   , ai ratti è   stato somministrato per via orale R406 a dosi fino a 100 mg/kg/die inclusi (o il suo profarmaco R788 a dosi fino a 100 mg/kg/die inclusi, ridotti a 50 mg/die). kg/die per le donne dopo aver superato la MTD), un forte inibitore Syk, due volte al giorno per 28 giorni.
    • In addition to standard toxicological assessments     , flow cytometric immunophenotyping and humoral immune response tests were performed by measuring levels of  anti   –   KLH  IgM    and IgG antibodies   .  Other immunotoxicity studies included three host resistance models in female Balb / c mice to further determine the effects of R406 on innate and acquired immunity.
    • After treatment with R406,   the expected immunomodulatory effects (e.g. decreased thymus and spleen weight, loss of bone marrow cells and decreased lymphocyte counts, including T and B lymphocytes) were observed in rat studies. .
    • These changes generally resolved during the 14-day treatment-free recovery period. KLH    challenge    in    rats  showed   no adverse effects on IgG or IgM   response       . R788 / 406, administered orally at doses up to and including 80 mg / kg / day for 28 days, did not affect bacterial or viral clearance in mouse resistance models of Listeria, streptococcus or influenza hosts, respectively.
    • This was related to previous in vitro macrophage and neutrophil function tests (evaluation of migratory ions, phagocytosis, oxidative burst and bactericidal activity) which showed that  R406   did not adversely affect macrophage or neutrophil function in immune responses. innate.
    • Taken together, these results show that   R406   has minimal functional immunotoxicity despite its lymphocytopenic effects, suggesting that Syk inhibition may not lead to unacceptable mechanism-based adverse effects.

 

To test whether IL-10 acts as an autocrine or paracrine growth factor for Ly-1 B cells, we treated mice continuously from birth to 8 weeks of age with a rat   monoclonal IgM antibody    that  specifically neutralizes      IL-10. murine. Mice treated in this manner were deprived of   peritoneal Ly-1 B cells, contained significantly reduced serum levels of immunoglobulin M, and were unable to generate  significant in vivo antibody responses   to intraperitoneal injection of alpha 1,3-dextran or phosphorylcholine,  antigens   , for which specific B cells are found in the subset of Ly-1 B cells.

In contrast, spleen B cells from  conventional anti   -IL-10-treated mice were normal with respect to total number, phenotype, and in vitro response to B cell mitogens and     trinitrophenyl hemocyanin antigen (TNP-   KLH )   from thymus-dependent keyhole patella. ).

The mechanism of Ly-1 B cell depletion appears to be related to increased levels of endogenous interferon gamma (IFN-gamma) in mice treated with anti-IL-10 because they   co-administered   anti-IFN-gamma sub anti   –   IL. – 10 neutralizing  antibodies    .  alli restored the number of Ly-1 B cells residing in the peritoneum of these mice.

In contrast, spleen B cells from  conventional anti   -IL-10-treated mice were normal with respect to total number, phenotype, and in vitro response to B cell mitogens and     trinitrophenyl hemocyanin antigen (TNP-   KLH )   from thymus-dependent keyhole patella. ).

The mechanism of Ly-1 B cell depletion appears to be related to increased levels of endogenous interferon gamma (IFN-gamma) in mice treated with anti-IL-10 because they   co-administered   anti-IFN-gamma sub anti   –   IL. – 10 neutralizing  antibodies    .  alli restored the number of Ly-1 B cells residing in the peritoneum of these mice. These results suggest that IL-10 is a growth regulator of Ly-1 B cells and identify a procedure for specific depletion of Ly-1 B cells, thus allowing further evaluation of the role of these cells in the immune system.

Anti-Flag antibody (KLH)

STJ98077 100 µl 280.8 EUR

Anti-BNP antibody (KLH)

STJ97879 100 µl 280.8 EUR

Anti-BNP antibody (KLH)

STJ97880 100 µl 280.8 EUR

Anti-BNP antibody (KLH)

STJ97881 100 µl 280.8 EUR

Anti-Calnexin antibody (KLH)

STJ97891 100 µl 280.8 EUR

Anti-Calregulin antibody (KLH)

STJ97892 100 µl 280.8 EUR

Anti-cAMP antibody (KLH)

STJ97893 100 µl 280.8 EUR

Anti-CD20 antibody (KLH)

STJ97914 100 µl 280.8 EUR

Anti-Clenbuterol antibody (KLH)

STJ97960 100 µl 280.8 EUR

Bovine Immunoglobulin M (IgM) ELISA Kit

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Bovine Immunoglobulin M (IgM) ELISA Kit

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Chicken Immunoglobulin M (IgM) ELISA Kit

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Chicken Immunoglobulin M (IgM) ELISA Kit

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Human Immunoglobulin M (IgM) ELISA Kit

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Human Immunoglobulin M (IgM) ELISA Kit

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Mouse Immunoglobulin M (IgM) ELISA Kit

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Mouse Immunoglobulin M (IgM) ELISA Kit

DLR-IgM-Mu-96T 96T 633.6 EUR

Rabbit Immunoglobulin M (IgM) ELISA Kit

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Rabbit Immunoglobulin M (IgM) ELISA Kit

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Bovine Immunoglobulin M (IgM) ELISA Kit

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Bovine Immunoglobulin M (IgM) ELISA Kit

RD-IgM-b-96Tests 96 Tests 750 EUR

Bovine Immunoglobulin M (IgM) ELISA Kit

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Bovine Immunoglobulin M (IgM) ELISA Kit

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Chicken Immunoglobulin M (IgM) ELISA Kit

RDR-IgM-Ch-48Tests 48 Tests 524.4 EUR

Chicken Immunoglobulin M (IgM) ELISA Kit

RDR-IgM-Ch-96Tests 96 Tests 723.6 EUR

Human Immunoglobulin M (IgM) ELISA Kit

RDR-IgM-Hu-48Tests 48 Tests 493.2 EUR

Human Immunoglobulin M (IgM) ELISA Kit

RDR-IgM-Hu-96Tests 96 Tests 678 EUR

Mouse Immunoglobulin M (IgM) ELISA Kit

RDR-IgM-Mu-48Tests 48 Tests 502.8 EUR

Mouse Immunoglobulin M (IgM) ELISA Kit

RDR-IgM-Mu-96Tests 96 Tests 693.6 EUR

Rabbit Immunoglobulin M (IgM) ELISA Kit

RDR-IgM-Rb-48Tests 48 Tests 640.8 EUR

Rabbit Immunoglobulin M (IgM) ELISA Kit

RDR-IgM-Rb-96Tests 96 Tests 890.4 EUR

Human Anti-Keyhole limpet hemocyanin (KLH) IgM ELISA Kit, 96 tests, Quantitative

700-145-KLM 1 Kit 927.6 EUR

Mouse Anti-Keyhole limpet hemocyanin (KLH) IgM ELISA Kit, 96 tests, Quantitative

700-155-VAM 1 Kit 854.4 EUR

Monkey Anti-Keyhole limpet hemocyanin (KLH) IgM ELISA Kit, 96 tests, Quantitative

700-180-KLM 1 Kit 927.6 EUR

Canine Anti-Keyhole limpet hemocyanin (KLH) IgM ELISA Kit, 96 tests, Quantitative

700-190-KLM 1 Kit 927.6 EUR

Rat anti Mouse IgM

41R-1116 100 ug 318 EUR

Mouse anti Rat IgM

41R-1122 100 ug 318 EUR

Mouse anti Rat IgM

41R-1123 100 ug 318 EUR

Rabbit anti Rat IgM

40C-CB1264 2 mg 342 EUR

Goat anti Rat IgM

40C-CB1271 1 mg 435.6 EUR

Rat(anti-CMV IgM)

QY-E11747 96T 448.8 EUR

Human keyhole limpet hemocyanin(KLH) antibody(IgM) ELISA Kit

CSB-E16534h-24T 1 plate of 24 wells 198 EUR

Human keyhole limpet hemocyanin(KLH) antibody(IgM) ELISA Kit

1-CSB-E16534h
  • 1080.00 EUR
  • 6571.20 EUR
  • 3480.00 EUR
  • 1 plate of 96 wells
  • 10 plates of 96 wells each
  • 5 plates of 96 wells each

Mouse keyhole limpet hemocyanin (KLH) antibody (IgM) ELISA kit

CSB-E13375m-24T 1 plate of 24 wells 198 EUR

Mouse keyhole limpet hemocyanin (KLH) antibody (IgM) ELISA kit

1-CSB-E13375m
  • 1080.00 EUR
  • 6571.20 EUR
  • 3480.00 EUR
  • 1 plate of 96 wells
  • 10 plates of 96 wells each
  • 5 plates of 96 wells each

Rat KLH ELISA Kit

ERK0075 96Tests 625.2 EUR

Lyophilized Exosome Standards

ExoA375 100 μg 364.2 EUR

Lyophilized Exosome Standards

ExoHT29 100 μg 364.2 EUR

Lyophilized Exosome Standards

ExoMCF7 100 μg 364.2 EUR

Lyophilized Exosome Standards

ExoPC3 100 μg 364.2 EUR

Lyophilized Exosome Standards

ExoPLASMA 100 μg 364.2 EUR

Lyophilized Exosome Standards

ExoUrine 100 μg 364.2 EUR

Density Standards 6ml

DEN1012 PK10 69.31 EUR

Chlorine meter standards

WAT1076 EACH 269.04 EUR

Human Anti-Keyhole limpet hemocyanin (KLH) IgG+IgM ELISA Kit, 96 tests, Quantitative

700-135-KGM 1 kit 927.6 EUR

Bovine/Cow Anti-Keyhole limpet hemocyanin (KLH) IgM ELISA Kit, 96 tests, Quantitative

700-205-KLM 1 Kit 854.4 EUR

Anti-Myc-tag antibody (KLH)

STJ90105 200 µl 236.4 EUR

Anti-His-tag antibody (KLH)

STJ90106 200 µl 236.4 EUR

Anti-HA-tag antibody (KLH)

STJ90107 200 µl 236.4 EUR

Anti-FLAG-tag antibody (KLH)

STJ90108 200 µl 236.4 EUR

Anti-E alpha antibody (KLH)

STJ98053 100 µl 280.8 EUR

Anti-Ki-67 antibody (KLH)

STJ98194 100 µl 280.8 EUR

Anti-Ki-67 antibody (KLH)

STJ98195 100 µl 280.8 EUR

Anti-PARP-1 antibody (KLH)

STJ98308 100 µl 280.8 EUR

Anti-Actin beta antibody (KLH)

STJ97814 100 µl 280.8 EUR

Anti-Bcl-2 antibody (KLH)

STJ97860 100 µl 280.8 EUR

Custom production of antibodies in 5 Rats using customer supplied antigen (std 63 days protocol)

RAT-5 1 1365.6 EUR

Rabbit anti Rat IgM (FITC)

43C-CB1265 1.5 mg 346.8 EUR

Rabbit anti Rat IgM (rhodamine)

43C-CB1266 1.5 mg 363.6 EUR

Rabbit anti Rat IgM (biotin)

43C-CB1268 1.5 mg 441.6 EUR

Rabbit anti Rat IgM (HRP)

43C-CB1269 1.5 mg 478.8 EUR

Goat anti Rat IgM (FITC)

43C-CB1272 1 mg 436.8 EUR

Goat anti Rat IgM (rhodamine)

43C-CB1273 1 mg 482.4 EUR

Goat anti Rat IgM (biotin)

43C-CB1275 1 mg 466.8 EUR

Goat anti Rat IgM (HRP)

43C-CB1276 1 mg 534 EUR

Mouse anti Rat IgM (HRP)

43R-1632 100 ug 338.4 EUR

Mouse anti Rat IgM (HRP)

43R-1633 100 ug 338.4 EUR

Goat anti-Rat IgM Antibody

abx401442-05mg 0.5 mg 444 EUR

Goat Anti-Rat IgM-AP

LF-SA8011AP 1.0 ml 423.6 EUR

Goat Anti-Rat IgM-BIOT

LF-SA8011B 1.0 mg 374.4 EUR

Goat Anti-Rat IgM-FITC

LF-SA8011F 1.0 mg 328.8 EUR

Goat Anti-Rat IgM-HRP

LF-SA8011H 1.0 ml 336 EUR

Goat Anti-Rat IgM-PE

LF-SA8011P 0.5 mg 524.4 EUR

Goat Anti-Rat IgM-TXRD

LF-SA8011T 1.0 mg 294 EUR

Goat Anti-Rat IgM-TRITC

LF-SA8011TR 1.0 mg 319.2 EUR

Goat Anti-Rat IgM-UNLB

LF-SA8011U 1.0 mg 313.2 EUR

Chloramphenicol [KLH]

DAG2974K 1mg 1170 EUR

Amoxicillin [KLH]

DAG4477 0.5mg 1248 EUR

Ampicillin [KLH]

DAG4478 0.5mg 1248 EUR

Apramycin [KLH]

DAG4479 0.5mg 1248 EUR

Bacitracin [KLH]

DAG4480 0.5ml 1170 EUR

Cloxacillin [KLH]

DAG4483 0.5mg 1248 EUR

Gentamicin [KLH]

DAG4484 1mg 1248 EUR

Neomycin [KLH]

DAG4486 0.5ml 1248 EUR

Streptomycin [KLH]

DAG4488 0.5ml 1482 EUR

Melamine [KLH]

DAG4492 1mg 1170 EUR

Ractopamine [KLH]

DAGA-001K 1mg 1170 EUR

Salbutamol [KLH]

DAGA-002K 1mg 1248 EUR

Clenbuterol [KLH]

DAGA-003K 1mg 1248 EUR

Pb2+ [KLH]

DAGA-005K 1mg 1560 EUR

Cd2+ [KLH]

DAGA-006K 1mg 1263.6 EUR

Hg2+ [KLH]

DAGA-007K 1mg 1263.6 EUR

Cu2+ [KLH]

DAGA-008K 1mg 1326 EUR

Ochratoxin [KLH]

DAGA-012K 1mg 1326 EUR

Zearalenone [KLH]

DAGA-013K 1mg 1014 EUR

Fumonisin [KLH]

DAGA-014K 1mg 1201.2 EUR

Deoxynivalenol [KLH]

DAGA-016K 500ul 1482 EUR

Sterigmatocystin [KLH]

DAGA-017K 1mg 1326 EUR

Methyltestosterone [KLH]

DAGA-020K 1mg 1248 EUR

Testosterone [KLH]

DAGA-021K 1mg 1232.4 EUR

Medroxyprogesterone [KLH]

DAGA-022K 1mg 1263.6 EUR

Progesterone [KLH]

DAGA-023K 1mg 1326 EUR

Hexadecadrol [KLH]

DAGA-024K 1mg 1248 EUR

Diethylstilbestrol [KLH]

DAGA-025K 1mg 1326 EUR

Hexoestrolum [KLH]

DAGA-026K 1mg 1248 EUR

Hydrocortisone [KLH]

DAGA-027K 1mg 1248 EUR

Estradiol [KLH]

DAGA-028K 1mg 1170 EUR

Nonylphenol [KLH]

DAGA-032K 1mg 1248 EUR

Ribavirin [KLH]

DAGA-033K 1mg 1294.8 EUR

Amantadine [KLH]

DAGA-034K 1mg 1294.8 EUR

Sulfadiazine [KLH]

DAGA-035K 1mg 1294.8 EUR

Sulfaquinoxaline [KLH]

DAGA-036K 1mg 1170 EUR

Sulfamethazine [KLH]

DAGA-037K 1mg 1014 EUR

Sulfadimethoxine [KLH]

DAGA-038K 1mg 1326 EUR

Trimethoprim [KLH]

DAGA-039K 1mg 1326 EUR

Chloramphenicol [KLH]

DAGA-040K 1mg 1248 EUR

Florfenicol [KLH]

DAGA-042K 1mg 1014 EUR

Kanamycin [KLH]

DAGA-044K 1mg 1170 EUR

Neomycin [KLH]

DAGA-045K 1mg 1326 EUR

Lincomycin [KLH]

DAGA-046K 1mg 1482 EUR

Pirlimycin [KLH]

DAGA-047K 1mg 1326 EUR

Erythrocin [KLH]

DAGA-048K 1mg 1014 EUR

Spectinomycin [KLH]

DAGA-049K 1mg 1294.8 EUR

Avermectin [KLH]

DAGA-050K 1mg 1248 EUR

Tylosin [KLH]

DAGA-051K 1mg 1326 EUR

Tilmicosin [KLH]

DAGA-052K 1mg 1482 EUR

Vancomycin [KLH]

DAGA-053K 1mg 1248 EUR

Salinomycin [KLH]

DAGA-054K 1mg 1482 EUR

Monensin [KLH]

DAGA-055K 1mg 1248 EUR

Albendazole [KLH]

DAGA-056K 1mg 1324.8 EUR

Natamycin [KLH]

DAGA-057K 1mg 1248 EUR

Metronidazole [KLH]

DAGA-058K 1mg 1248 EUR

Tetracycline [KLH]

DAGA-059K 1mg 1014 EUR

Olaquindox [KLH]

DAGA-060K 1mg 1326 EUR

Ciprofloxacin [KLH]

DAGA-063K 1mg 1404 EUR

Sarafloxacin [KLH]

DAGA-064K 1mg 1294.8 EUR

Ofloxacin [KLH]

DAGA-065K 1mg 1294.8 EUR

Enrofloxacin [KLH]

DAGA-066K 1mg 1248 EUR

Ceftiofur [KLH]

DAGA-068K 1mg 1404 EUR

Cefquinome [KLH]

DAGA-069K 1mg 1232.4 EUR

Fluoroquinolones [KLH]

DAGA-070K 1mg 1014 EUR

Sulfanilamides [KLH]

DAGA-071K 1mg 1014 EUR

Biotin [KLH]

DAGA-074K 1mg 1248 EUR

Erythrosine [KLH]

DAGA-076K 1mg 1248 EUR

Chrysoidine [KLH]

DAGA-077K 1mg 1248 EUR

Tartrazine [KLH]

DAGA-079K 1mg 1232.4 EUR

Sudan [KLH]

DAGA-081K 1mg 1326 EUR

Carbendazim [KLH]

DAGA-082K 1mg 1404 EUR

Chlorothalonil [KLH]

DAGA-083K 1mg 1248 EUR

Chlorpyrifos [KLH]

DAGA-085K 1mg 1248 EUR

Carbaryl [KLH]

DAGA-086K 1mg 1248 EUR

Acetochlor [KLH]

DAGA-087K 1mg 1248 EUR

Acetamiprid [KLH]

DAGA-088K 1mg 1248 EUR

Imidacloprid [KLH]

DAGA-089K 1mg 1170 EUR

Fenpropathrin [KLH]

DAGA-090K 1mg 1170 EUR

Pentachlorophenol [KLH]

DAGA-093K 1mg 1170 EUR

Nitrazepam [KLH]

DAGA-097K 1mg 1404 EUR

Clonazepam [KLH]

DAGA-098K 1mg 1248 EUR

Diazepam [KLH]

DAGA-099K 1mg 1248 EUR

Dihydrochlorothiazide [KLH]

DAGA-100K 1mg 1248 EUR

Benzothiazinone [KLH]

DAGA-101K 1mg 1248 EUR

Benzothiazine [KLH]

DAGA-102K 1mg 1248 EUR

Mercaptobenzothiazole [KLH]

DAGA-104K 1mg 1092 EUR

Imazalil [KLH]

DAGA-105K 1mg 1201.2 EUR

Benzophenone [KLH]

DAGA-106K 1mg 1248 EUR

Acrylamide [KLH]

DAGA-110K 1mg 1248 EUR

Gossypol [KLH]

DAGA-118K 1mg 1248 EUR

KLH antibody

70R-30550 1 ml 224.4 EUR

KLH Antibody

48158-100ul 100ul 399.6 EUR

KLH Antibody

48158-50ul 50ul 286.8 EUR
×

These results suggest that IL-10 is a growth regulator of Ly-1 B cells and identify a procedure for specific depletion of Ly-1 B cells, thus allowing further evaluation of the role of these cells in the immune system.

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