Compositions comprising an antibody and camostat mesylate (cm)
A technology of camostat mesylate and a composition, applied in the field of compositions comprising antibodies and camostat mesylate (CM), can solve the problem of high cost, inconvenience and pain, reducing patient compliance, etc. question
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Embodiment 1
[0040] Example 1: Intrinsic instability of a panel of monoclonal antibodies in simulated intestinal fluid (SIF)
[0041] Simulated Intestinal Fluid (SIF) is based on TNO-TIM (TM) The intestinal model system was formulated using the same formulation, but substantially in scaled down volume, as detailed below.
[0042] Preparation of SIF
[0043] The bile solution was prepared by slowly adding 2.0 g (+ / - 0.02 g) of bile powder to 250 g (+ / - 5 g) of purified water with constant stirring until a clear solution was obtained.
[0044] The trypsin solution was prepared by adding 2.1 g (+ / - 0.2 g) of trypsin powder to 150 g (+ / - 3 g) of purified water. Use a mixer, and handle with care to minimize foaming. Once a homogeneous mixture was obtained, the solution was centrifuged at 3500 rpm for 20 minutes and the supernatant was stored on ice.
[0045] Small Intestinal Electrolyte Solution (SIES) 25% (concentrated) is made by adding purified water to 250g (+ / -5g) sodium chloride, 3...
Embodiment 2
[0058] Example 2: Stabilization of Monoclonal Antibodies Using Camostat Mesylate
[0059]The panel of mAbs studied in Example 1 was also incubated in SIF containing camostat mesylate (CM, Sequoia Research Products) to determine whether inhibition of proteases would improve mAb stability. Add CM to the electrolyte solution described above to a concentration of 350 mg / ml (CM is high concentration but below saturation point) and warm to 50°C to dissolve. CM was added to the SIF / mAb to a final concentration of 10 mg / ml. The time points used and analysis by SDS-PAGE were as described in Example 1. Anti-IL23 was incubated in SIF with CM as described in Example 1 under different conditions from the other antibodies.
[0060] CM stabilized the monoclonal antibody when added at 10 mg / ml. The half-life of each mAb studied was extended to over 21 hours, as shown in Figure 2(b). The gel shown in Figure 2(a) shows the stability of the anti-IL6 mAbs, which is representative of the sta...
Embodiment 3
[0061] Example 3: Binding of CM-stabilized monoclonal antibodies to their target ligands
[0062] The monoclonal antibodies were incubated in SIF in the absence and presence of CM, as shown in Examples 1 and 2, and the ability of the monoclonal antibodies to bind to their ligands was determined using ELISA. In short, the Nunc Maxisorp (TM) Plates were coated overnight with ligands (IL-6 and IL-23 in this example). Plates were washed and blocked with bovine serum albumin. SIF samples were diluted and added to the plate along with standard curve mAbs and incubated at room temperature to allow binding. The plate was washed and a peroxidase-conjugated anti-human Fc region antibody was added to the wells. After incubation, the plate was washed and incubated with OPD substrate to obtain a colorimetric signal. The reaction was stopped with sulfuric acid and the absorbance was read at 490nm.
[0063] Binding of anti-IL6 (Fig. 3(a)) and anti-IL23 (Fig. 3(b)) to their correspondi...
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