STABILIZATION OF VIRAL COMPOSITIONS
This invention concerns stabilized virus compositions, preferably a herpesvirus which may be an attenuated or genetically modified herpes simplex virus or varicella zoster virus, and a method of stabilizing viruses and immunizing preparations by the addition of sugars, preferably glucose and amino acids, preferably lysine.
1 . A live virus preparation comprising:
(a) a herpes virus,
(b) lysine at a concentration of 0.2 to 200 g/l, and
(c) a sugar or sugar alcohol at 0.2 to 200 g/l.
2 . The preparation of claim 1 , wherein the sugar or sugar alcohol is glucose.
3 . A live virus preparation prepared by lyophilizing the preparation of claim 1 .
4 . A process for preparing stabilized vaccines containing attenuated live viruses, said process comprising contacting attenuated live viruses with an effective amount of the stabilizing agent of claim 1 .
5 . The process of claim 4 wherein said live virus is a herpes virus.
6 . The process of claim 4 wherein said herpes virus is a herpes simplex type 2.
7 . The process of claim 6 wherein said herpes virus is a herpes simplex type 2 wherein the PK domain has been deleted.
8 . The process of claim 4 , wherein said contacting is conducted prior to a lyophilization step.
9 . A stabilizing agent for live herpes virus viral immunizing agent comprising lysine and glucose.
10 . The stabilizing agent of claim 9 wherein said immunizing agent is a herpes virus type 2.
11 . The stabilizing agent of claim 10 wherein said herpes virus is a herpes simplex type 2 in which the PK domain has been deleted.
12 . A method of preparing a live virus immunizing agent that comprises mixing a live herpes virus with a stabilizer comprising an aqueous solution of a sugar or sugar alcohol at 1-100 g/l and lysine at 2 to 20 g/l.
13 . The method of claim 1 wherein said live virus is a herpes simplex type 2 virus in which the PK domain has been deleted.
14 . A method of harvesting live herpes virus for use in a live virus immunizing agent that includes the step of disrupting cells containing said virus in the presence of a stabilizer comprising an aqueous solution of a sugar or sugar alcohol at 1-200 g/l and lysine at 0.2 to 200 g/l.
15 . The method of claim 14 wherein said sugar or sugar alcohol is glucose.
16 . The method of claim 14 wherein said live herpes virus is a herpes simplex type 2 virus.
17 . The method of claim 16 wherein said herpes virus is a herpes simplex type 2 in which the PK domain has been deleted.
18 . A method of harvesting herpes simplex virus for use in a live virus immunizing agent that includes the step of separating the growth medium from the cells in which the herpes simplex is grown and adding a stabilizer to the growth medium containing excreted virus comprising lysine and sugar or sugar alcohol to provide a final concentration of the sugar or sugar alcohol at 1-200 g/l and of the lysine at 0.2 to 200 g/l.
19 . The method of claim 18 wherein said sugar or sugar alcohol is glucose.
20 . The method of claim 18 wherein said herpes virus is herpes simplex type 2.
21 . The method of claim 20 wherein said herpes virus is a herpes simplex type 2 in which the PK domain has been deleted.
22 . A live virus preparation according to claim 1 , wherein said preparation comprises lysine at a concentration selected from 0.3M, 0.5M, 0.7M, and 0.9M.
23 . A live virus preparation according to claim 1 , wherein said preparation comprises lysine is at a concentration of 0.3M.
24 . A live virus preparation according to claim 1 , wherein said preparation comprises 2% glucose and lysine at a concentration of 0.3M.
25 . A live virus preparation according to claim 1 , wherein said herpes virus is herpes simplex type I.
26 . A live virus preparation according to claim 1 , wherein said herpes virus is herpes simplex type II.
27 . A live virus preparation according to claim 1 , wherein said herpes virus is varicella zoster virus.
28 . A live virus preparation according to claim 1 , wherein said herpes virus is herpes simplex type II wherein the PK domain has been deleted.
29 . A live virus preparation according to claim 1 , wherein said herpes virus is Herpes Simplex Virus-2 ICP10ΔPK(CS).
30 . A live virus preparation according to claim 1 , wherein said preparation is not buffered.
31 . A live virus preparation according to claim 1 , wherein said preparation is supplied for storage at a temperature selected from −20 degree C., −4 degree C., 23 degree C., and room temperature.
32 . A live virus preparation according to claim 1 , wherein said preparation is supplied for storage as a low temperature frozen product.
33 . A live virus preparation according to claim 1 , wherein said preparation is supplied for storage as a lyophilized preparation.
34 . A live virus preparation according to claim 32 or 33 , wherein said preparation is prepared from a liquid composition.