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

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-Calnexin antibody (KLH)
STJ97891	100 µl	234 EUR
Anti-Calregulin antibody (KLH)
STJ97892	100 µl	234 EUR
Anti-cAMP antibody (KLH)
STJ97893	100 µl	234 EUR
Anti-CD20 antibody (KLH)
STJ97914	100 µl	234 EUR
Anti-Clenbuterol antibody (KLH)
STJ97960	100 µl	234 EUR
Anti-Flag antibody (KLH)
STJ98077	100 µl	234 EUR
Anti-MTAP Antibody (KLH)
STJ503536	100 µg	586 EUR
Mouse Keyhole Limpet Hemocyanin (KLH IgM) ELISA Kit
abx053480-96tests	96 tests	668 EUR
Human Anti-Meningococcal Group A/C/W/Y Oligosaccharides IgM (Combo) ELISA Kit, 96 tests, Quantitative
600-884-IGM	1 Kit	1017 EUR
Human Anti-Keyhole limpet hemocyanin (KLH) IgM ELISA Kit, 96 tests, Quantitative
700-145-KLM	1 Kit	773 EUR
Mouse Anti-Keyhole limpet hemocyanin (KLH) IgM ELISA Kit, 96 tests, Quantitative
700-155-VAM	1 Kit	712 EUR
Monkey Anti-Keyhole limpet hemocyanin (KLH) IgM ELISA Kit, 96 tests, Quantitative
700-180-KLM	1 Kit	773 EUR
Canine Anti-Keyhole limpet hemocyanin (KLH) IgM ELISA Kit, 96 tests, Quantitative
700-190-KLM	1 Kit	773 EUR
Rat(anti-CMV IgM)
QY-E11747	96T	374 EUR
Rabbit anti Rat IgM
40C-CB1264	2 mg	285 EUR
Goat anti Rat IgM
40C-CB1271	1 mg	363 EUR
Rat anti Mouse IgM
41R-1116	100 ug	265 EUR
Mouse anti Rat IgM
41R-1122	100 ug	265 EUR
Mouse anti Rat IgM
41R-1123	100 ug	265 EUR
Bovine Immunoglobulin M (IgM) ELISA Kit
RD-IgM-b-48Tests	48 Tests	453 EUR
Bovine Immunoglobulin M (IgM) ELISA Kit
RD-IgM-b-96Tests	96 Tests	625 EUR
Bovine Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-b-48T	48T	456 EUR
Bovine Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-b-96T	96T	590 EUR
Chicken Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-Ch-48T	48T	426 EUR
Chicken Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-Ch-96T	96T	549 EUR
Human Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-Hu-48T	48T	404 EUR
Human Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-Hu-96T	96T	518 EUR
Mouse Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-Mu-48T	48T	411 EUR
Mouse Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-Mu-96T	96T	528 EUR
Rabbit Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-Rb-48T	48T	508 EUR
Rabbit Immunoglobulin M (IgM) ELISA Kit
DLR-IgM-Rb-96T	96T	661 EUR
Bovine Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-b-48Tests	48 Tests	473 EUR
Bovine Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-b-96Tests	96 Tests	654 EUR
Chicken Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-Ch-48Tests	48 Tests	437 EUR
Chicken Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-Ch-96Tests	96 Tests	603 EUR
Human Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-Hu-48Tests	48 Tests	411 EUR
Human Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-Hu-96Tests	96 Tests	565 EUR
Mouse Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-Mu-48Tests	48 Tests	419 EUR
Mouse Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-Mu-96Tests	96 Tests	578 EUR
Rabbit Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-Rb-48Tests	48 Tests	534 EUR
Rabbit Immunoglobulin M (IgM) ELISA Kit
RDR-IgM-Rb-96Tests	96 Tests	742 EUR
Mouse keyhole limpet hemocyanin (KLH) antibody (IgM) ELISA kit
CSB-E13375m-24T	1 plate of 24 wells	165 EUR
Mouse keyhole limpet hemocyanin (KLH) antibody (IgM) ELISA kit
1-CSB-E13375m	900.00 EUR 5476.00 EUR 2900.00 EUR	1 plate of 96 wells 10 plates of 96 wells each 5 plates of 96 wells each
Human keyhole limpet hemocyanin(KLH) antibody(IgM) ELISA Kit
CSB-E16534h-24T	1 plate of 24 wells	165 EUR
Human keyhole limpet hemocyanin(KLH) antibody(IgM) ELISA Kit
1-CSB-E16534h	900.00 EUR 5476.00 EUR 2900.00 EUR	1 plate of 96 wells 10 plates of 96 wells each 5 plates of 96 wells each
Lyophilized Exosome Standards
ExoA375	100 μg	303.5 EUR
Lyophilized Exosome Standards
ExoHT29	100 μg	303.5 EUR
Lyophilized Exosome Standards
ExoMCF7	100 μg	303.5 EUR
Lyophilized Exosome Standards
ExoPC3	100 μg	303.5 EUR
Lyophilized Exosome Standards
ExoPLASMA	100 μg	303.5 EUR
Lyophilized Exosome Standards
ExoUrine	100 μg	303.5 EUR
Anti-Actin beta antibody (KLH)
STJ97814	100 µl	234 EUR
Anti-Bcl-2 antibody (KLH)
STJ97860	100 µl	234 EUR
Anti-E alpha antibody (KLH)
STJ98053	100 µl	234 EUR
Anti-Ki-67 antibody (KLH)
STJ98194	100 µl	234 EUR
Anti-Ki-67 antibody (KLH)
STJ98195	100 µl	234 EUR
Anti-PARP-1 antibody (KLH)
STJ98308	100 µl	234 EUR
Anti-Myc-tag antibody (KLH)
STJ90105	200 µl	197 EUR
Anti-His-tag antibody (KLH)
STJ90106	200 µl	197 EUR
Anti-HA-tag antibody (KLH)
STJ90107	200 µl	197 EUR
Anti-FLAG-tag antibody (KLH)
STJ90108	200 µl	197 EUR
Human Anti-Keyhole limpet hemocyanin (KLH) IgG+IgM ELISA Kit, 96 tests, Quantitative
700-135-KGM	1 kit	773 EUR
Bovine/Cow Anti-Keyhole limpet hemocyanin (KLH) IgM ELISA Kit, 96 tests, Quantitative
700-205-KLM	1 Kit	712 EUR
Rat KLH ELISA Kit
ERK0075	96Tests	521 EUR
Goat anti-Rat IgM Antibody
abx401442-05mg	0.5 mg	370 EUR
Goat Anti-Rat IgM-AP
LF-SA8011AP	1.0 ml	353 EUR
Goat Anti-Rat IgM-BIOT
LF-SA8011B	1.0 mg	312 EUR
Goat Anti-Rat IgM-FITC
LF-SA8011F	1.0 mg	274 EUR
Goat Anti-Rat IgM-HRP
LF-SA8011H	1.0 ml	280 EUR
Goat Anti-Rat IgM-PE
LF-SA8011P	0.5 mg	437 EUR
Goat Anti-Rat IgM-TXRD
LF-SA8011T	1.0 mg	245 EUR
Goat Anti-Rat IgM-TRITC
LF-SA8011TR	1.0 mg	266 EUR
Goat Anti-Rat IgM-UNLB
LF-SA8011U	1.0 mg	261 EUR
Rabbit anti Rat IgM (FITC)
43C-CB1265	1.5 mg	289 EUR
Rabbit anti Rat IgM (rhodamine)
43C-CB1266	1.5 mg	303 EUR
Rabbit anti Rat IgM (biotin)
43C-CB1268	1.5 mg	368 EUR
Rabbit anti Rat IgM (HRP)
43C-CB1269	1.5 mg	399 EUR
Goat anti Rat IgM (FITC)
43C-CB1272	1 mg	364 EUR
Goat anti Rat IgM (rhodamine)
43C-CB1273	1 mg	402 EUR
Goat anti Rat IgM (biotin)
43C-CB1275	1 mg	389 EUR
Goat anti Rat IgM (HRP)
43C-CB1276	1 mg	445 EUR
Mouse anti Rat IgM (HRP)
43R-1632	100 ug	282 EUR
Mouse anti Rat IgM (HRP)
43R-1633	100 ug	282 EUR
Custom production of antibodies in 5 Rats using customer supplied antigen (std 63 days protocol)
RAT-5	1	1138 EUR
Polyclonal Goat anti-GST α-form
GST-ANTI-1	50 uL	280 EUR
Polyclonal Goat anti-GST μ-form
GST-ANTI-2	50 uL	280 EUR
Polyclonal Goat anti-GST p-form
GST-ANTI-3	50 uL	280 EUR
Chloramphenicol [KLH]
DAG2974K	1mg	975 EUR
Amoxicillin [KLH]
DAG4477	0.5mg	1040 EUR
Ampicillin [KLH]
DAG4478	0.5mg	1040 EUR
Apramycin [KLH]
DAG4479	0.5mg	1040 EUR
Bacitracin [KLH]
DAG4480	0.5ml	975 EUR
Cloxacillin [KLH]
DAG4483	0.5mg	1040 EUR
Gentamicin [KLH]
DAG4484	1mg	1040 EUR
Neomycin [KLH]
DAG4486	0.5ml	1040 EUR
Streptomycin [KLH]
DAG4488	0.5ml	1235 EUR
Melamine [KLH]
DAG4492	1mg	975 EUR
Ractopamine [KLH]
DAGA-001K	1mg	975 EUR
Salbutamol [KLH]
DAGA-002K	1mg	1040 EUR
Clenbuterol [KLH]
DAGA-003K	1mg	1040 EUR
Pb2+ [KLH]
DAGA-005K	1mg	1300 EUR
Cd2+ [KLH]
DAGA-006K	1mg	1053 EUR
Hg2+ [KLH]
DAGA-007K	1mg	1053 EUR
Cu2+ [KLH]
DAGA-008K	1mg	1105 EUR
Ochratoxin [KLH]
DAGA-012K	1mg	1105 EUR
Zearalenone [KLH]
DAGA-013K	1mg	845 EUR
Fumonisin [KLH]
DAGA-014K	1mg	1001 EUR
Deoxynivalenol [KLH]
DAGA-016K	500ul	1235 EUR
Sterigmatocystin [KLH]
DAGA-017K	1mg	1105 EUR
Methyltestosterone [KLH]
DAGA-020K	1mg	1040 EUR
Testosterone [KLH]
DAGA-021K	1mg	1027 EUR
Medroxyprogesterone [KLH]
DAGA-022K	1mg	1053 EUR
Progesterone [KLH]
DAGA-023K	1mg	1105 EUR
Hexadecadrol [KLH]
DAGA-024K	1mg	1040 EUR
Diethylstilbestrol [KLH]
DAGA-025K	1mg	1105 EUR
Hexoestrolum [KLH]
DAGA-026K	1mg	1040 EUR
Hydrocortisone [KLH]
DAGA-027K	1mg	1040 EUR
Estradiol [KLH]
DAGA-028K	1mg	975 EUR
Nonylphenol [KLH]
DAGA-032K	1mg	1040 EUR
Ribavirin [KLH]
DAGA-033K	1mg	1079 EUR
Amantadine [KLH]
DAGA-034K	1mg	1079 EUR
Sulfadiazine [KLH]
DAGA-035K	1mg	1079 EUR
Sulfaquinoxaline [KLH]
DAGA-036K	1mg	975 EUR
Sulfamethazine [KLH]
DAGA-037K	1mg	845 EUR
Sulfadimethoxine [KLH]
DAGA-038K	1mg	1105 EUR
Trimethoprim [KLH]
DAGA-039K	1mg	1105 EUR
Chloramphenicol [KLH]
DAGA-040K	1mg	1040 EUR
Florfenicol [KLH]
DAGA-042K	1mg	845 EUR
Kanamycin [KLH]
DAGA-044K	1mg	975 EUR
Neomycin [KLH]
DAGA-045K	1mg	1105 EUR
Lincomycin [KLH]
DAGA-046K	1mg	1235 EUR
Pirlimycin [KLH]
DAGA-047K	1mg	1105 EUR
Erythrocin [KLH]
DAGA-048K	1mg	845 EUR
Spectinomycin [KLH]
DAGA-049K	1mg	1079 EUR
Avermectin [KLH]
DAGA-050K	1mg	1040 EUR
Tylosin [KLH]
DAGA-051K	1mg	1105 EUR
Tilmicosin [KLH]
DAGA-052K	1mg	1235 EUR
Vancomycin [KLH]
DAGA-053K	1mg	1040 EUR
Salinomycin [KLH]
DAGA-054K	1mg	1235 EUR
Monensin [KLH]
DAGA-055K	1mg	1040 EUR
Albendazole [KLH]
DAGA-056K	1mg	1104 EUR
Natamycin [KLH]
DAGA-057K	1mg	1040 EUR
Metronidazole [KLH]
DAGA-058K	1mg	1040 EUR
Tetracycline [KLH]
DAGA-059K	1mg	845 EUR
Olaquindox [KLH]
DAGA-060K	1mg	1105 EUR
Ciprofloxacin [KLH]
DAGA-063K	1mg	1170 EUR
Sarafloxacin [KLH]
DAGA-064K	1mg	1079 EUR
Ofloxacin [KLH]
DAGA-065K	1mg	1079 EUR
Enrofloxacin [KLH]
DAGA-066K	1mg	1040 EUR
Ceftiofur [KLH]
DAGA-068K	1mg	1170 EUR
Cefquinome [KLH]
DAGA-069K	1mg	1027 EUR
Fluoroquinolones [KLH]
DAGA-070K	1mg	845 EUR
Sulfanilamides [KLH]
DAGA-071K	1mg	845 EUR
Biotin [KLH]
DAGA-074K	1mg	1040 EUR
Erythrosine [KLH]
DAGA-076K	1mg	1040 EUR
Chrysoidine [KLH]
DAGA-077K	1mg	1040 EUR
Tartrazine [KLH]
DAGA-079K	1mg	1027 EUR
Sudan [KLH]
DAGA-081K	1mg	1105 EUR
Carbendazim [KLH]
DAGA-082K	1mg	1170 EUR
Chlorothalonil [KLH]
DAGA-083K	1mg	1040 EUR
Chlorpyrifos [KLH]
DAGA-085K	1mg	1040 EUR
Carbaryl [KLH]
DAGA-086K	1mg	1040 EUR
Acetochlor [KLH]
DAGA-087K	1mg	1040 EUR
Acetamiprid [KLH]
DAGA-088K	1mg	1040 EUR
Imidacloprid [KLH]
DAGA-089K	1mg	975 EUR
Fenpropathrin [KLH]
DAGA-090K	1mg	975 EUR
Pentachlorophenol [KLH]
DAGA-093K	1mg	975 EUR
Nitrazepam [KLH]
DAGA-097K	1mg	1170 EUR
Clonazepam [KLH]
DAGA-098K	1mg	1040 EUR
Diazepam [KLH]
DAGA-099K	1mg	1040 EUR
Dihydrochlorothiazide [KLH]
DAGA-100K	1mg	1040 EUR
Benzothiazinone [KLH]
DAGA-101K	1mg	1040 EUR
Benzothiazine [KLH]
DAGA-102K	1mg	1040 EUR
Mercaptobenzothiazole [KLH]
DAGA-104K	1mg	910 EUR
Imazalil [KLH]
DAGA-105K	1mg	1001 EUR
Benzophenone [KLH]
DAGA-106K	1mg	1040 EUR
Acrylamide [KLH]
DAGA-110K	1mg	1040 EUR
Gossypol [KLH]
DAGA-118K	1mg	1040 EUR
KLH Antibody
48158-100ul	100ul	333 EUR
KLH Antibody
48158-50ul	50ul	239 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.