In vitro: Tocilizumab inhibits the binding of IL-6 to its receptors, and thus reduces the cytokines pro-inflammatory activity by competing for both the soluble and membrane-bound forms of the human IL-6 receptor. The inhibitory profile of anti-IL-6R mAb Tocilizumab is independent of membrane-bound IL-6R expression. Tocilizumab has anticancer potency via apoptosis induction as an agonistic IL-6R regulator. It has also been demonstrated that tocilizumab has an anti-proliferative effect on glioma cells via inhibition of the JAK-STAT3 pathway. Tocilizumab exhibits a significant growth inhibition in NSCLC cells (H460, A549, H1299 and H358), with proliferation significantly decreased by approximately 40% in A549 cells. Tocilizumab does not alter the levels of the ERK1/2, STAT3, NFκB and phosphorylated ERK1/2 and STAT3 proteins, but this antibody does considerably increase the expression of phosphorylated NFκB in NSCLC cells. Tocilizumab significantly inhibits expression of both IL-8 and MMP-9, known as the major angiogenic factors.
In vivo: A series of clinical studies has shown that inhibition of IL-6 signaling by tocilizumab is therapeutically effective in rheumatoid arthritis, juvenile idiopathic arthritis, Castleman's disease, and Crohn's disease. In all of these diseases, tocilizumab ameliorates inflammatory manifestations, and normalizes acute phase protein levels. Tocilizumab as a monotherapy and in combination, such as with methotrexate in case of rheumatoid arthritis, seems to be well tolerated. Tocilizumab markedly decreases the number of invaded capillary vessels in tumors.
Research (Wash D C). 2022 Jul 19;2022:9826426.
Programming of Regulatory T Cells In Situ for Nerve Regeneration and Long-Term Patency of Vascular Grafts
Tocilizumab purchased from AbMole
|Cell lines||non-small cell lung cancer (NSCLC) cells (A549, H460, H358 and H1299 cells)|
|Preparation method||Ten microliters of tocilizumab, MTX or 5-FU are added to 96-well plates containing 104 cells per well in 100 µl medium. The final concentrations of tocilizumab are 10, 100 and 1000 ng/ml. The final concentrations of MTX and 5-FU are 50 and 25 µg/ml, respectively. Following a 24-h incubation, WST-1 solution is added, and the optical density is analyzed at reference wavelengths of 450 and 620 nm.|
|Concentrations||10, 100 and 1000 ng/ml|
|Incubation time||24 h|
|Animal models||Male CB17/ICR-scid/scid mice (SCID mice)|
|Storage||-80°C for long term|
|Body Surface Area (m2)||0.007||0.025||0.15||0.05||0.02||0.5|
|Animal A (mg/kg) = Animal B (mg/kg) multiplied by||Animal B Km|
|Animal A Km|
For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.
 Peng Jiang, et al. Exp Ther Med. Intracisternal administration of an interleukin-6 receptor antagonist attenuates surgery-induced cognitive impairment by inhibition of neuroinflammatory responses in aged rats
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