Compositions and methods for treating viral infections
A composition, virus technology, applied in the field of compositions and methods for treating viral infections, capable of solving problems such as maintenance difficulties, non-strict compliance with cold chain procedures, equipment failure and power outages
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Embodiment 1
[0113] Example 1: Reverse Melt Preparation of Antigen-Containing Vesicles
[0114] This example describes a reverse-melt formulation for the preparation of antigen-containing nonionic surfactant vesicles (NISV). In Step 1, a 5:4:1 molar ratio of the following lipids was placed in a 50 ml flat-bottomed glass beaker: 1-monopalmitin (MPG), cholesterol (CHO), and dicetyl phosphate (DCP), Make sure that no powder gets on the sides of the glass beaker. Melt the lipid at about 120 to 125°C for 10 minutes in a heated oil bath with occasional stirring in a glass oven covered with aluminum foil.
[0115] At this stage, the inactivated antigen vaccine (reconstituted according to the manufacturer's Sanofi Pasteur vaccine) stock solution in a heated water bath at about 30 to 35° C. for 5 to 10 minutes. In step 2, the resulting vaccine stock solution was homogenized at 8000 rpm at 30 to 35 °C, and the molten lipid mixture was added to the homogenized vaccine stock solution (to obtain 6....
Embodiment 2
[0116] Example 2: Study on thermal stability of antigen-containing vesicles prepared by reverse melting method
[0117] To assess thermal stability, NISV was prepared as described in Example 1, and lyophilized aliquots were stored at two different thermal storage temperatures (5±3°C and 40±2°C) (prior to reconstitution). ). If stored in a refrigerator at 2°C to 8°C, the freeze-dried Vaccines are stable; reconstituted vaccines are unstable and should be used immediately. in lyophilized or reconstituted form The vaccine is also stable at elevated temperatures. At specific times, stability samples were removed from the thermostat, reconstituted in WFI and analyzed for appearance, pH, microscopy, Zeta potential, nano size and ELISA (antigen content). The vaccine control (unformulated lyophilized Vaccine) (Test 7 (TA7)), stored as above but without the addition of NISV, was also tested.
[0118] Rabies virus antigen content in NISV preparations was determined by performi...
Embodiment 3
[0131] Example 3: In vivo animal testing of antigen-containing vesicles prepared by the reverse melting method
[0132] Female Balb / C mice (6 to 8 weeks old; body weight 18 to 28 grams, Charles River Canada Inc.) were immunized once (n=8) intramuscularly on day 0 (with 0.1 ml of the indicated vaccine samples). Serum samples collected before and after the first immunization were analyzed for formulated and unformulated Humoral immune response to vaccines. The humoral immune response was determined by performing an IgG ELISA serology assay. An indirect ELISA was performed to assess anti-rabies virus-specific IgG titers in immune sera. Briefly, rabies virus antigen ( Vaccine, Sanofi Pasteur) coated each well of a 96-well ELISA plate overnight at 4°C. The next day the plates were washed with PBS containing 0.05% Tween 20 and then blocked with PBS containing 10% goat serum (1 hour at 37°C). After the incubation time, the plates were washed six times in wash buffer (0.05% Twee...
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