Inactivation of extracellular enveloped viruses in blood and blood components by phenthiazin-5-ium dyes plus light

• US 5,545,516 A
• Filed: 02/18/1994
• Issued: 08/13/1996
• Est. Priority Date: 05/01/1990
• Status: Expired due to Term

• Current Assignee: American National Red Cross
• Original Assignee: American National Red Cross
• Inventors: Wagner, Stephen J.
• Primary Examiner(s): Knode, Marian C.
• Assistant Examiner(s): SAUCIER, SANDRA E
• Application Number:
  US08/199,344
• Time in Patent Office: 907 Days
• Field of Search: 435/2, 424/533, 424/529-532
• US Class Current: 435/2
• CPC Class Codes: A61K 35/16 Blood plasma; Blood serum umbilical cord blood A61K35/51
• CPC Class Codes: A61K 35/18 Erythrocytes haemoglobin A61K38/42
• CPC Class Codes: A61K 35/19 Platelets; Megacaryocytes
• CPC Class Codes: A61K 41/17 by ultraviolet [UV] or infrared [IR] light, X-rays or gamma rays
• CPC Class Codes: C12N 2795/12063 by chemical treatment
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

This invention provides methods for inactivating pathogenic contaminants in whole blood, plasma, cellular blood components, or in any combination thereof, by adding a phenthiazin-5-ium dye(s) thereto and irradiating said dye-containing composition for an effective length of time with light of wavelengths from 560 to 800 nm or red light, of an effective intensity, whereby the irradiation in conjunction with the dye(s) inactivate substantially all pathogenic contaminants contained therein. The methods of this invention inactivate pathogenic contaminants, such as viruses, bacteria and parasites, without substantially altering the whole blood, plasma, cellular blood components, or combinations thereof, such that they are suitable for transfusion.

Claims

1. A method for inactivating extracellular enveloped viruses which are present in a leukocyte-containing transfusible composition, comprising:

(a) leukodepleting said composition by a factor of at least 5 log₁₀ ;

(b) adding an adequate amount of at least one phenothiazin-5-ium dye to said leukodepleted composition of step (a), to produce therein an effective concentration of said dye(s), wherein said concentration is acceptable for transfusion and wherein said concentration in conjunction with light is effective to inactivate extracellular enveloped viruses contained in said composition; and

(c) irradiating said phenothiazin-5-ium dye-containing composition of step (b) for an effective length of time, with light of effective intensity and wavelengths which are absorbed by said dye(s), whereby said irradiation in conjunction with said dye(s) inactivate extracellular enveloped viruses in said composition.

9. The method of any one of claims 1-4, wherein said phenthiazin-5-ium dye is selected from the group consisting of methylene blue, toluidine blue O, azure A, azure B, azure C and thionine.

10. The method of claim 9, wherein said dye is methylene blue.

11. The method of claim 9, wherein said dye is toluidine blue O.

12. The method of claim 9, wherein said dye is thionine.

13. The method of claim 9, wherein said phenthiazin-5-ium dye is U.S.P. grade.

14. The method of any one of claims 1-4, wherein said effective inactivating concentration of the phenthiazin-5-ium dye(s) is from 0.2 μ

M to 50 μ

M.

15. The method of any one of claims 1-4, wherein said effective inactivating concentration of the phenthiazin-5-ium dye(s) is from 1 μ

M to 25 μ

M.

16. The method of any one of claims 1-4, wherein said effective inactivating concentration of the phenthiazin-5-ium dye(s) is from 1 μ

M to 5 μ

M.

17. The method of any one of claims 1-4, wherein said effective length of irradiation time is from 1 minute to 120 minutes.

18. The method of any one of claims 1-4, wherein said effective length of irradiation time is from 90 minutes to 120 minutes.

19. The method of any one of claims 1-4, wherein said extracellular enveloped viruses are selected from the group consisting of RNA viruses and DNA viruses.

20. The method of claim 19, wherein said enveloped RNA virus is a retrovirus.

21. The method of claim 20, wherein said extracellular retrovirus is human immunodeficiency virus -1.

22. The method of claim 19, wherein said RNA virus is bacteriophage φ

6.

23. The method of claim 19, wherein said extracellular enveloped DNA virus is selected from the group consisting of:

herpes viruses;

hepatitis B virus;

pox viruses; and

cytomegaloviruses.

24. The method of claim 19, wherein said extracellular enveloped DNA virus is pseudorabies virus.

25. The method of claim 19, wherein said extracellular enveloped RNA virus is selected from the group consisting of:

hepatitis A virus;

hepatitis C virus;

paramyxoviruses; and

toga viruses.

26. The method of claim 19, wherein said extracellular enveloped RNA virus is Sindbis virus.

27. The method of any one of claims 1, 3 or 4, wherein said extracellular enveloped virus is vesicular stomatitis virus.

28. The method of any one of claims 1-3, wherein said leukocyte-containing composition contains ARC-8.

29. The method of any one of claims 1-3, wherein said leukocyte-containing composition is whole blood.

30. The method of any one of claims 1-3, wherein said leukocyte-containing composition contains plasma.

2. A method for inactivating extracellular enveloped viruses which are present in a leukocyte-containing, red blood cell-containing composition, comprising:

(a) leukodepleting said composition;

(b) adding an adequate amount of at least one phenothiazin-5-ium dye to said composition to produce therein an effective concentration of said dye(s), wherein said concentration is acceptable for transfusion and wherein said concentration in conjunction with light is effective to inactivate extracellular enveloped viruses contained in said composition; and

(c) irradiating said phenothiazin-5-ium dye-containing composition for an effective length of time, with light of an effective intensity and of wavelengths which are absorbed by said dye(s), whereby said irradiation in conjunction with said dye(s) inactivate extracellular enveloped viruses in said composition, wherein said red blood cells retain their viability.

31. The method of either one of claims 2 or 3, wherein the resulting leukodepleted composition is a transfusible composition of step (c).

3. A method for inactivating extracellular enveloped viruses which are present in a leukocyte-containing, platelet-containing composition, comprising:

(a) leukodepleting said composition;

(b) adding an adequate amount of at least one phenothiazin-5-ium dye to said composition to produce therein an effective concentration of said dye(s), wherein said concentration is acceptable for transfusion and wherein said concentration in conjunction with light is effective to inactivate extracellular enveloped viruses contained in said composition; and

(c) irradiating said phenothiazin-5-ium dye-containing composition for an effective length of time, with light of an effective intensity and of wavelengths which are absorbed by said dye(s) whereby said irradiation in conjunction with said dye(s) inactivate extracellular enveloped viruses in said composition, wherein said platelets retain their viability.

4. A method for inactivating extracellular enveloped viruses which are present in leukocyte-containing plasma, comprising:

(a) leukodepleting said plasma by a factor of at least 5 log₁₀ ;

(b) adding an adequate amount of at least one phenothiazin-5-ium dye to said leukodepleted plasma of step (a) to produce therein an effective concentration of said dye(s), wherein said concentration is acceptable for transfusion and wherein said concentration in conjunction with light is effective to inactivate extracellular enveloped viruses contained in said composition; and

(c) irradiating said phenothiazin-5-ium dye-containing plasma of step (b) for an effective length of time, with light of an effective intensity and of wavelengths which are absorbed by said dye(s), whereby said irradiation in conjunction with said dye(s) inactivate extracellular enveloped viruses in said plasma.

5. The method of claim 4, wherein said plasma contains ARC-8.

6. The method of claim 4, wherein said plasma is transfusible.

7. The method of claim 4, wherein the plasma concentration is from 2.5% to 100%.

8. The method of claim 4, wherein the plasma concentration is from 50% to 100%.

32. A process for treating a leukocyte-containing red blood cell-containing composition in order to inactivate any extracellular lipid enveloped human pathogenic virus contained therein without incurring substantial disruption or inactivation of said cells, comprising:

(a) leukodepleting said composition;

(b) contacting said red blood cell-containing composition having a concentration of red blood cells≧

1×

10⁹ cells/ml with a viricidally effective amount of at least one phenothiazin-5-ium dye, with light and with ambient oxygen to inactivate said virus if present therein and to effect a virus kill of at least 4 log₁₀.

33. A process according to claim 32, wherein extracellular lipid enveloped virus is inactivated.