阅读说明：本技术 灭活脂包膜病毒的环境相容性去污剂 (Environmentally compatible detergents for inactivation of lipoenveloped viruses ) 是由 J-B·法西 J·金德曼 B·蒂尔 T·R·克雷尔 于 2018-10-30 设计创作，主要内容包括：本发明涉及使用环境相容性去污剂灭活脂包膜病毒的方法,并且涉及使用环境相容性去污剂制备生物药物的方法。本发明还提供了环境相容性去污剂。(The present invention relates to a method for inactivating a lipid-enveloped virus using an environmentally compatible detergent, and to a method for preparing a biopharmaceutical using an environmentally compatible detergent. The invention also provides environmentally compatible detergents.)

1. a compound of the following formula (VIII):

in the formula (I), the compound is shown in the specification,

r represents a hydrocarbon group having a straight chain of 2 to 12 carbon atoms and one or more methyl groups as substituents on the straight chain;

m represents an integer of 1 to 4; and

a represents a polyoxyethylene residue.

2. A compound according to claim 1, wherein a represents a polyoxyethylene residue comprising 4 to 16 oxyethylene units, preferably a polyoxyethylene residue comprising 9 or 10 oxyethylene units.

3. The compound of claim 1 or 2, wherein m is equal to 1.

4. A compound according to any one of claims 1 to 3, wherein R represents a hydrocarbon group having a straight chain of 2 to 6 carbon atoms and one or more methyl groups as substituents on the straight chain.

5. A compound according to any one of claims 1 to 4, wherein R represents a hydrocarbon group having a straight chain of 2 to 6 carbon atoms and 2 to 4 methyl groups as substituents on the straight chain.

6. A compound according to any one of claims 1 to 5, wherein R represents a hydrocarbon group having a straight chain of4 carbon atoms and 4 methyl groups as substituents on the straight chain.

7. A compound according to any one of claims 1 to 6, wherein R represents 2,4, 4-trimethyl-pentan-2-yl.

8. The compound of claim 1, wherein the compound of formula (VIII) is the following compound:

wherein m and z are integers independently selected from the group consisting of:

m is 1 to 4, and m is a linear chain,

and z is 1 to 5.

9. The compound of claim 8, wherein m is equal to 1.

10. The compound of claim 1, wherein the compound of formula (VIII) is the following compound:

where n is an integer from 4 to 16, preferably n is equal to 9 or 10.

11. A compound according to any one of claims 1 to 10, with the proviso that 29- [4- (1,1,3,3-tetramethylbutyl) phenyl ] -3,6,9,12,15,18,21,24, 27-nonaxononacosan-1-ol of the formula:

12. a method of inactivating a virus having a lipid envelope, wherein the method comprises the steps of:

a) adding a detergent to a liquid, preparing a mixture of said detergent and said liquid; and

b) incubating the mixture to inactivate the virus;

wherein the detergent is polyoxyethylene ether, and the detergent is a non-phenolic detergent.

13. The method of claim 12, wherein the detergent is environmentally compatible.

14. The method of claim 12 or 13, wherein the detergent is a non-ionic detergent.

15. The method of any one of claims 12 to 14, wherein the detergent is a compound of any one of claims 1 to 11.

16. The method of any one of claims 12 to 14, wherein the detergent is a polyoxyethylene alkyl ether.

17. The method of claim 16, wherein the polyoxyethylene alkyl ether is a polyoxyethylene cycloalkyl ether.

18. The method of claim 17, wherein the cycloalkyl moiety of the polyoxyethylene cycloalkyl ether is an alkyl-substituted cycloalkyl moiety.

19. The method of claim 18, wherein the alkyl-substituted cycloalkyl moiety is a branched alkyl-substituted cycloalkyl moiety.

20. The process of any one of claims 17 to 19, wherein the polyoxyethylene cycloalkyl ether is polyoxyethylene cyclohexyl ether.

21. The method of any one of claims 17 to 20, wherein the polyoxyethylene cycloalkyl ether is not a heterocyclic polyoxyethylene cycloalkyl ether.

22. The method of any one of claims 12-14 and 16-21, wherein the detergent has the following structure according to formula (I):

wherein x, y and z are integers independently selected from the group consisting of:

x is 0 to 5, and x is,

y is 0 to 5, and y is,

and z is 0 to 20.

23. The method of claim 22, wherein the detergent has the following structure according to formula (II):

24. the method of claim 23, wherein the detergent has the following structure according to formula (III):

wherein n is an integer of4 to 16.

25. The method of claim 24, wherein n is equal to 9 or 10.

26. The method of claim 22, wherein the detergent has the following structure according to formula (IV):

27. the method of claim 26, wherein the detergent has the following structure according to formula (V):

wherein n is an integer of4 to 16.

28. The method of claim 27, wherein n is equal to 9 or 10.

29. The method of any one of claims 12 to 14 and 16, wherein the detergent is a linear polyoxyethylene alkyl ether.

30. The method of claim 29, wherein the detergent is a linear polyoxyethylene cetyl ether.

31. The method of claim 30, wherein the detergent has the following structure according to formula (VI):

wherein x is equal to 15.

32. The method of claim 31, wherein the detergent has the following structure according to formula (VII):

wherein x is equal to 15 and n is an integer from 5 to 15.

33. The method of claim 32, wherein n is equal to 10.

34. The method of any one of claims 12 to 33, wherein the detergent is suitable for inactivating the virus.

35. The method according to any one of claims 12 to 34, wherein in step a) no organic solvent is added to the liquid.

36. The method of any one of claims 12 to 34, wherein step a) further comprises adding a solvent to the liquid, and in step a) a solvent/detergent mixture for inactivating the virus is prepared by adding the detergent and the solvent to the liquid.

37. The method of claim 36, wherein the solvent is an organic solvent.

38. A process according to claim 36 or 37, wherein the solvent is tri-n-butyl phosphate.

39. The method according to any one of claims 12 to 38, wherein in step a) no further detergent other than said detergent is added.

40. The method of any one of claims 12 to 38, wherein no detergent other than said detergent is added in the method.

41. The method of any one of claims 12 to 38, wherein step a) further comprises adding further detergent to the liquid.

42. The method of claim 41, wherein the other detergent is polysorbate 80.

43. The method of any one of claims 12 to 42, wherein the liquid comprises a biopharmaceutical product.

44. The method of any one of claims 12 to 43, wherein the liquid comprises a biologic drug.

45. The method of claim 44, wherein the biologic is not a viral vaccine.

46. The method of any one of claims 44 or 45, wherein the biological agent is a blood factor, an immunoglobulin such as a monoclonal antibody, a surrogate enzyme, a vaccine, a gene therapy vector, a growth factor or a growth factor receptor.

47. The method of any one of claims 44 to 46, wherein the biopharmaceutical is a therapeutic protein.

48. The method of any one of claims 44-47, wherein the biologic is a blood factor that is factor I (fibrinogen), factor II (prothrombin), tissue factor, factor V, factor VII, or factor VIIa, factor VIII, factor IX, factor X, factor XI, factor XII, factor XIII, Von Willebrand Factor (VWF), prekallikrein, High Molecular Weight Kininogen (HMWK), fibronectin, antithrombin III, heparin cofactor II, protein C, protein S, protein Z, plasminogen, α 2-antiplasmin, tissue plasminogen activator (tPA), urokinase, plasminogen activator inhibitor-1 (PAI1), or plasminogen activator inhibitor-2 (PAI 2).

49. The method according to any one of claims 44 to 48, wherein the biopharmaceutical is factor VIII, preferably recombinant human factor VIII.

50. The method of any one of claims 44 to 47, wherein the biopharmaceutical is an immunoglobulin which is an immunoglobulin or monoclonal antibody from human plasma.

51. The method of any one of claims 12 to 50, wherein prior to step a) or between steps a) and b), the method further comprises the step of filtering the liquid or mixture with a depth filter.

52. The method of any one of claims 12 to 51, wherein in step b) the mixture is incubated for at least 1 hour.

53. The method of any one of claims 12 to 52, wherein in step b) the mixture is incubated at a temperature of 0 to 10 ℃ or the mixture is incubated at a temperature of 16 to 25 ℃.

54. The method according to any one of claims 44-53, wherein the method further comprises, after step b), the step of:

c) purifying the biopharmaceutical.

55. The method of claim 54, wherein said purifying comprises separating said biopharmaceutical from said detergent.

56. The method of claim 54 or 55, wherein the purifying comprises separating the biopharmaceutical from the other detergent.

57. The method of any one of claims 54 to 56, wherein the purification of the biopharmaceutical comprises purifying the biopharmaceutical by at least one chromatographic purification.

58. The method according to any one of claims 54 to 57, wherein the at least one chromatographic purification is by anion exchange chromatography and/or by cation exchange chromatography.

59. A method of preparing a biopharmaceutical, wherein the method comprises the method of any of claims 44 to 58, the biopharmaceutical being as in any of claims 44 to 58.

60. The method of claim 59, wherein the method further comprises the step of preparing a pharmaceutical formulation comprising the biopharmaceutical after the method of any of claims 44-58.

61. Use of the detergent of any one of claims 12 to 34 in a method of inactivating a virus having a lipid envelope.

62. The use of claim 61, wherein no detergent other than the detergent is used in the use.

63. The use according to claim 61 or 62, wherein, in said use, no organic solvent is used.

64. The use of claim 61, wherein the method of inactivating the virus is a method using a solvent/detergent treatment comprising the use of a detergent of any one of claims 12 to 34.

65. The use according to any one of claims 61 to 64, wherein the viral inactivation is viral inactivation of a fluid comprising the biopharmaceutical of any one of claims 44 to 50.

66. A composition comprising the detergent of any one of claims 12 to 34.

67. A detergent according to any one of claims 12 to 34.

68. The composition of claim 66, wherein the composition further comprises a biopharmaceutical according to any of claims 44 to 50.

69. The composition of claim 66 or 68, wherein the composition does not comprise any organic solvent.

70. The composition of claim 66 or 68, wherein the composition further comprises an organic solvent of any one of claims 37 and 38.

71. The composition of any one of claims 66 or 68 to 70, wherein said composition does not comprise any other detergent than said detergent.

72. The composition of any one of claims 66 or 68 to 70, wherein the composition further comprises an additional detergent according to any one of claims 41 and 42.

73. A kit for virus inactivation, wherein the kit comprises the detergent of claim 67 or the composition of any one of the preceding claims and further comprises a chromatographic resin for chromatographic purification of any one of claims 57 to 58.

74. The kit of claim 73, wherein the kit further comprises a depth filter.

75. A process for the synthesis of a compound of formula (VIII),

in the formula (I), the compound is shown in the specification,

r represents a hydrocarbon group having a straight chain of 2 to 12 carbon atoms and one or more methyl groups as substituents on the straight chain;

m represents an integer of 1 to 4; and

a represents a polyoxyethylene residue;

wherein the method comprises the steps of:

A) converting a phenol of formula (IX) to an alcohol of formula (X):

wherein, R is the same as that described above,

wherein R and m are as described above;

and

(B) the alcohol of formula (X) is converted to the polyoxyethylene ether of formula (VIII) as described above.

76. A process for the synthesis of a compound of formula (VIII),

in the formula (I), the compound is shown in the specification,

r represents a hydrocarbon group having a straight chain of 2 to 12 carbon atoms and one or more methyl groups as substituents on the straight chain;

m represents an integer of 1 to 4; and

a represents a polyoxyethylene residue;

wherein the method comprises the steps of:

(1) reacting toluene to obtain substituted toluene of the following formula (XI), wherein R is as described above,

(2) converting the substituted toluene of formula (XI) into a compound of formula (XII) wherein R and m are as defined above and X is selected from the group consisting of a hydroxyl group, a bromine atom, an iodine atom and a chlorine atom,

and

(3) the compound of formula (XII) is converted into the polyoxyethylene ether of formula (VIII) as described above.

77. A process for the synthesis of a compound of formula (VIIIa),

in the formula (I), the compound is shown in the specification,

r represents a hydrocarbon group having a straight chain of 2 to 12 carbon atoms and one or more methyl groups as substituents on the straight chain; and

a represents a polyoxyethylene residue;

wherein the method comprises the steps of:

(I) converting benzyl alcohol to a compound of formula (XIII) wherein R is as defined above,

and

(II) converting the compound of formula (XIII) to a polyoxyethylene ether of formula (VIIIa) as described above.

78. The method of any one of claims 75 to 77, wherein A represents a polyoxyethylene residue comprising 4 to 16 oxyethylene units.

79. The method of claim 78, wherein A represents a polyoxyethylene residue comprising 8 to 10 oxyethylene units.

80. The method of claim 78, wherein A represents a polyoxyethylene residue comprising 9 or 10 oxyethylene units.

81. The method of any one of claims 75, 76, and 78-80, wherein m is equal to 1.

82. A process according to any one of claims 75 to 81, wherein R represents a hydrocarbyl group having a straight chain of 2 to 6 carbon atoms and one or more methyl groups as substituents on the straight chain.

83. A process according to any one of claims 75 to 82, wherein R represents a hydrocarbyl group having a straight chain of 2 to 6 carbon atoms and 2 to 4 methyl groups as substituents on the straight chain.

84. A process according to any one of claims 75 to 83, wherein R represents a hydrocarbyl group having a straight chain of4 carbon atoms and 4 methyl groups as substituents on the straight chain.

85. A process according to any one of claims 75 to 84, wherein R represents 2,4, 4-trimethyl-pentan-2-yl.

86. A process according to any one of claims 75 to 77, wherein the compound of formula (VIII) is the following:

wherein m and z are integers independently selected from the group consisting of:

m is 1 to 4;

and z is 1 to 5.

87. The method of claim 86, wherein m is equal to 1.

88. A process according to any one of claims 75 to 77, wherein the compound of formula (VIII) is the following:

wherein n is an integer from 4 to 16, preferably n is equal to 9 or 10.

89. The process of any one of claims 76 and 78 to 88, wherein the conversion in step (2) is a free radical reaction using AIBN (azobisisobutyronitrile) as a free radical initiator.

90. The method of any one of claims 76 and 78 to 89, wherein X is a bromine atom.

91. The process according to any one of claims 76 and 78 to 90, wherein the conversion in step (2) uses N-bromosuccinimide (NBS) as a reagent.

92. The process of any one of claims 76 and 78 to 91, wherein the conversion in step (3) uses TBME (methyl tert-butyl ether) as solvent.

93. The process of any one of claims 76 and 78 to 92, wherein the conversion in step (3) is carried out for at least 2 hours, preferably at ambient temperature.

94. The process of any one of claims 76 and 78 to 93, wherein the conversion in step (3) is carried out for no more than 5 hours, preferably at ambient temperature.

95. The process of any one of claims 76 and 78 to 94, wherein the conversion in step (3) is carried out for 3 hours, preferably at ambient temperature.

96. The process of any one of claims 75 to 95, wherein said process is carried out on a scale yielding at least 100g, at least 1kg, at least 10kg, at least 100kg or at least 1000kg of said compound of formula (VIII).

Technical Field

The present invention relates to a method for inactivating lipid-enveloped (lipid-enveloped) viruses using an environmentally compatible detergent, and to a method for preparing biopharmaceutical drugs (biopharmaceutical drugs) using an environmentally compatible detergent. The invention also provides environmentally compatible detergents.

Background

The use of biopharmaceuticals is of increasing importance as a method of treating a number of diseases, disorders or conditions affecting the health of an individual. Typically, biopharmaceuticals are obtained by purification of biological fluids or recombinant production in host cells (e.g., mammalian cell lines). However, viral contamination is a serious problem in such biopharmaceutical manufacturing processes. Viral contamination may be introduced into the biopharmaceutical manufacturing process by the biological fluid to be purified or by the use of products of animal origin. Unlike bacterial contamination, viral contamination is difficult to detect. However, if infectious viruses are incorporated into biopharmaceutical formulations without attention to viral contamination, serious health risks are posed to the patient. Thus, viral inactivation is of crucial importance in biopharmaceutical production.

In many biopharmaceutical manufacturing processes, detergents are used for viral inactivation. Typically, these detergents are mixed with a solvent in a so-called solvent/detergent (S/D) treatment. The detergent Triton X-100 has been used for many years for S/D treatment of commercial products.

However, recent ecological studies have shown that Triton X-100 and its decomposition products may become endocrine disruptors of aquatic organisms, raising concerns from the viewpoint of environmental impact (see "ECHA Support document for identification of4- (1,1,3,3-tetramethylbutyl) phenol, ethoxylated as substention sodium high-purity phenol) with end-capping properties, while still utilizing basic nutrient benefits to the environment, and thus high-grade cellulose degradation of PBSs/salts of PBSs (CMHA 4-ethoxylated matter, CMHA 3-tetramethyl phosphate, PBSs 4-propyl phosphate, PBSs 4-tetramethyl phosphate, PBSs 4-tetramethyl phosphate, 1,3,3-tetramethylbutyl) phenol) that may have a severe impact on the environment, causing the same degree of concern as CMR and PBT/vPvB) ", 12 months and 12 days of 2012). Thus, there is a need for alternative environmentally compatible detergents for viral inactivation.

Disclosure of Invention

The present invention meets the above needs and solves the above problems in the art by providing the following embodiments:

the toxic activity of Triton-X100 is caused by a phenolic moiety that has the ability to harbor in certain endocrine receptors of marine organisms. Consistently, based on computer simulation predictions of endocrine disruptor activity, there is no evidence that the non-phenolic (non-phenolic) polyoxyethylene ether detergents of the present invention have endocrine disruptor activity.

The present inventors have surprisingly found that environmentally compatible non-phenolic polyoxyethylene ether detergents (e.g., Triton X-100Reduced, Triton N-101Reduced, and Brij C10) are effective in inactivating lipoenveloped viruses during S/D processing. The inventors have also synthesized environmentally compatible non-phenolic polyoxyethylene ether detergents that are effective in inactivating lipid-enveloped viruses in S/D and single-detergent (single-detergent) treatments. Accordingly, the inventors have found that environmentally compatible non-phenolic polyoxyethylene ether detergents can be used in the method of inactivating lipid-enveloped viruses of the present invention.

Accordingly, the present invention provides environmentally compatible non-phenolic polyoxyethylene ether detergents and improved methods of inactivating lipid-enveloped viruses by providing the following preferred embodiments:

1. a compound of the following formula (VIII):

wherein the content of the first and second substances,

r represents a hydrocarbon group having a straight chain of 2 to 12 carbon atoms and one or more methyl groups as substituents on the straight chain;

m represents an integer of 1 to 4; and

a represents a polyoxyethylene residue.

2. The compound according to item 1, wherein a represents a polyoxyethylene residue comprising 4 to 16 oxyethylene units, preferably a polyoxyethylene residue comprising 9 or 10 oxyethylene units.

3. The compound of item 1 or 2, wherein m is equal to 1.

4. The compound according to any one of claims 1 to 3, wherein R represents a hydrocarbon group having a straight chain of 2 to 6 carbon atoms and one or more methyl groups as substituents on the straight chain.

5. The compound according to any one of claims 1 to 4, wherein R represents a hydrocarbon group having a straight chain of 2 to 6 carbon atoms and 2 to 4 methyl groups as substituents on the straight chain.

6. The compound according to any one of claims 1 to 5, wherein R represents a hydrocarbon group having a straight chain of4 carbon atoms and 4 methyl groups as substituents on the straight chain.

7. A compound according to any one of claims 1 to 6, wherein R represents 2,4, 4-trimethyl-pentan-2-yl.

8. The compound according to item 1, wherein the compound of formula (VIII) is the following compound:

wherein m and z are integers independently selected from the group consisting of:

m is 1 to 4;

and z is 1 to 5.

9. The compound of claim 8, wherein m is equal to 1.

10. The compound according to item 1, wherein the compound of formula (VIII) is the following compound:

wherein n is an integer from 4 to 16, preferably wherein n is equal to 9 or 10.

11. A compound according to any one of claims 1 to 10, with the proviso that 29- [4- (1,1,3,3-tetramethylbutyl) phenyl ] -3,6,9,12,15,18,21,24, 27-nonaxononacosan-1-ol of the formula:

12. a method of inactivating a virus having a lipid envelope (lipid envelope), the method comprising the steps of:

a) adding a detergent to a liquid, preparing a mixture of said detergent and said liquid; and

b) incubating the mixture to inactivate the virus;

wherein the detergent is polyoxyethylene ether, and the detergent is a non-phenolic detergent.

13. The method of claim 12, wherein the detergent is environmentally compatible.

14. The method of claim 12 or 13, wherein the detergent is a non-ionic detergent.

15. The method according to any one of claims 12 to 14, wherein the detergent is a compound according to any one of claims 1 to 11.

16. The method of any of claims 12-14, wherein the detergent is a polyoxyethylene alkyl ether.

17. The method of claim 16, wherein the polyoxyethylene alkyl ether is a polyoxyethylene cycloalkyl ether.

18. The method of item 17, wherein the cycloalkyl moiety of the polyoxyethylene cycloalkyl ether is an alkyl-substituted cycloalkyl moiety.

19. The method of item 18, wherein the alkyl-substituted cycloalkyl moiety is a branched alkyl-substituted cycloalkyl moiety.

20. The method of any one of claims 17-19, wherein the polyoxyethylene cycloalkyl ether is polyoxyethylene cyclohexyl ether.

21. The method of any of claims 17-20, wherein the polyoxyethylene cycloalkyl ether is not a heterocyclic polyoxyethylene cycloalkyl ether.

22. The method of any of claims 12-14 and 16-21, wherein the detergent has the following structure according to formula (I):

wherein x, y and z are integers independently selected from the group consisting of:

x is 0 to 5;

y is 0 to 5;

and z is 0 to 20.

23. The method of claim 22, wherein the detergent has the following structure according to formula (II):

24. the method of item 23, wherein the detergent has the following structure according to formula (III):

wherein n is an integer of4 to 16.

25. The method of item 24, wherein n is equal to 9 or 10.

26. The method of claim 22, wherein the detergent has the following structure according to formula (IV):

27. the method of claim 26, wherein the detergent has the following structure according to formula (V):

wherein n is an integer of4 to 16.

28. The method of item 27, wherein n is equal to 9 or 10.

29. The method of any of claims 12-14 and 16, wherein the detergent is a linear polyoxyethylene alkyl ether.

30. The method of claim 29, wherein the detergent is a linear polyoxyethylene cetyl ether.

31. The method of item 30, wherein the detergent has the following structure according to formula (VI):

where x is equal to 15.

32. The method of item 31, wherein the detergent has the following structure according to formula (VII):

wherein x is equal to 15 and n is an integer from 5 to 15.

33. The method of item 32, wherein n equals 10.

34. The method of any of claims 12 to 33, wherein the detergent is suitable for inactivating the virus.

35. The method according to any one of claims 12 to 34, wherein in step a) no organic solvent is added to the liquid.

36. The method according to any of claims 12 to 34, wherein step a) further comprises adding a solvent to the liquid, and in step a) a solvent/detergent mixture for inactivating the virus is prepared by adding the detergent and the solvent to the liquid.

37. The method of claim 36, wherein the solvent is an organic solvent.

38. The method of claim 36 or 37, wherein the solvent is tri-n-butyl phosphate.

39. The method according to any one of claims 12 to 38, wherein in step a) no further detergent is added in addition to the detergent.

40. The method according to any one of claims 12 to 38, wherein no detergent other than the detergent is added in the method.

41. The method according to any one of claims 12 to 38, wherein step a) further comprises adding further detergent to the liquid.

42. The method of item 41, wherein the additional detergent is polysorbate 80.

43. The method of any of claims 12-42, wherein the liquid comprises a biopharmaceutical product.

44. The method of any one of claims 12 to 43, wherein the liquid comprises a biologic drug.

45. The method of item 44, wherein the biologic is not a viral vaccine.

46. The method of any one of claims 44 or 45, wherein the biological agent is a blood factor, an immunoglobulin (e.g., a monoclonal antibody), a surrogate enzyme, a vaccine, a gene therapy vector, a growth factor, or a growth factor receptor.

47. The method of any one of claims 44-46, wherein the biologic is a therapeutic protein.

48. The method of any one of claims 44-47, wherein the biologic is a blood factor that is factor I (fibrinogen), factor II (prothrombin), tissue factor, factor V, factor VII, or factor VIIa, factor VIII, factor IX, factor X, factor XI, factor XII, factor XIII, von Willebrand factor (VWF, von Willebrand factor), prekallikrein, High Molecular Weight Kininogen (HMWK), fibronectin, antithrombin III, heparin cofactor II, protein C, protein S, protein Z, plasminogen, α 2-antiplasmin, tissue plasminogen activator (tPA), urokinase, plasminogen activator inhibitor-1 (PAI1), or plasminogen activator inhibitor-2 (PAI 2).

49. The method of any one of claims 44 to 48, wherein the biopharmaceutical is factor VIII, preferably recombinant human factor VIII.

50. The method of any one of claims 44 to 47, wherein the biopharmaceutical is an immunoglobulin which is an immunoglobulin or monoclonal antibody from human plasma.

51. The method according to any one of claims 12 to 50, wherein prior to step a) or between steps a) and b), the method further comprises the step of filtering the liquid or mixture with a depth filter.

52. The method according to any one of claims 12 to 51, wherein in step b) the mixture is incubated for at least 1 hour.

53. The method according to any one of claims 12 to 52, wherein in step b) the mixture is incubated at a temperature of 0 to 10 ℃ or wherein the mixture is incubated at a temperature of 16 to 25 ℃.

54. The method according to any one of claims 44 to 53, wherein the method further comprises, after step b), the step of:

c) purifying the biopharmaceutical.

55. The method of item 54, wherein the purifying comprises separating the biopharmaceutical from the detergent.

56. The method of claim 54 or 55, wherein the purifying comprises separating the biopharmaceutical from the other detergent.

57. The method of any of claims 54-56, wherein said purifying of said biopharmaceutical comprises purifying said biopharmaceutical by at least one chromatographic purification.

58. The method according to any one of claims 54 to 57, wherein the at least one chromatographic purification is by anion exchange chromatography and/or by cation exchange chromatography.

59. A method of preparing a biopharmaceutical, the method comprising any of claims 44 to 58, wherein the biopharmaceutical is as in any of claims 44 to 58.

60. The method of claim 59, further comprising the step of preparing a pharmaceutical formulation comprising the biopharmaceutical after the method of any of claims 44-58.

61. Use of the detergent of any one of items 12 to 34 in a method of inactivating a virus having a lipid envelope.

62. The use of item 61, wherein in said use no detergent other than said detergent is used.

63. The use of item 61 or 62, wherein in said use, no organic solvent is used.

64. The use according to item 61, wherein the method for inactivating the virus is a method using a solvent/detergent treatment comprising the use of a detergent according to any one of items 12 to 34.

65. Use according to any of claims 61 to 64, wherein the viral inactivation is viral inactivation of a fluid comprising a biological agent according to any of claims 44 to 50.

66. A composition comprising the detergent of any one of claims 12 to 34.

67. A detergent according to any one of claims 12 to 34.

68. The composition of claim 66, wherein the composition further comprises the biopharmaceutical of any of claims 44-50.

69. The composition of claim 66 or 68, wherein the composition does not comprise any organic solvent.

70. The composition of any of claims 66 or 68, wherein the composition further comprises the organic solvent of any of claims 37 and 38.

71. The composition of any of claims 66 or 68-70, wherein the composition does not comprise any detergent other than the detergent.

72. The composition of any of claims 66 or 68 to 70, wherein the composition further comprises an additional detergent of any of 41 and 42.

73. A kit for virus inactivation, wherein the kit comprises the detergent of item 67 or the composition of any one of the preceding items, and further comprises a chromatographic resin for chromatographic purification of any one of items 57 to 58.

74. The kit of claim 73, wherein the kit further comprises a depth filter.

75. A process for the synthesis of a compound of formula (VIII),

wherein the content of the first and second substances,

r represents a hydrocarbon group having a straight chain of 2 to 12 carbon atoms and one or more methyl groups as substituents on the straight chain;

m represents an integer of 1 to 4; and

a represents a polyoxyethylene residue;

wherein the method comprises the steps of:

A) converting a phenol of formula (IX) to an alcohol of formula (X):

wherein, R is the same as that described above,

wherein R and m are as described above;

and

(B) the alcohol of formula (X) is converted into a polyoxyethylene ether having the general formula (VIII) as described above.

76. A process for the synthesis of a compound of formula (VIII),

wherein the content of the first and second substances,

r represents a hydrocarbon group having a straight chain of 2 to 12 carbon atoms and one or more methyl groups as substituents on the straight chain;

m represents an integer of 1 to 4; and

a represents a polyoxyethylene residue;

wherein the method comprises the steps of:

(1) reacting toluene to obtain substituted toluene of the following formula (XI), wherein R is as described above,

(2) converting the substituted toluene of formula (XI) into a compound of formula (XII) wherein R and m are as defined above and X is selected from the group consisting of a hydroxyl group, a bromine atom, an iodine atom and a chlorine atom,

and

(3) the compound of formula (XII) is converted into a polyoxyethylene ether having the general formula (VIII) as described above.

77. A process for the synthesis of a compound of formula (VIIIa),

wherein the content of the first and second substances,

r represents a hydrocarbon group having a straight chain of 2 to 12 carbon atoms and one or more methyl groups as substituents on the straight chain; and

a represents a polyoxyethylene residue;

wherein the method comprises the steps of:

(I) converting benzyl alcohol to a compound of formula (XIII) wherein R is as defined above,

and

(II) converting the compound of formula (XIII) into a polyoxyethylene ether having the general formula (VIIIa) as described above.

78. The method of any one of claims 75-77, wherein A represents a polyoxyethylene residue comprising 4 to 16 oxyethylene units.

79. The method of item 78, wherein A represents a polyoxyethylene residue comprising 8 to 10 oxyethylene units.

80. The method of item 78, wherein A represents a polyoxyethylene residue comprising 9 or 10 oxyethylene units.

81. The method of any of claims 75, 76, and 78-80, wherein m is equal to 1.

82. The process according to any one of claims 75 to 81, wherein R represents a hydrocarbon group having a straight chain of 2 to 6 carbon atoms and one or more methyl groups as substituents on the straight chain.

83. The process according to any one of claims 75 to 82, wherein R represents a hydrocarbon group having a straight chain of 2 to 6 carbon atoms and 2 to 4 methyl groups as substituents on the straight chain.

84. The process according to any one of claims 75 to 83, wherein R represents a hydrocarbon group having a straight chain of4 carbon atoms and 4 methyl groups as substituents on the straight chain.

85. The process according to any one of claims 75 to 84, wherein R represents 2,4, 4-trimethyl-pentan-2-yl.

86. The method according to any one of claims 75 to 77, wherein the compound of formula (VIII) is the following compound:

wherein m and z are integers independently selected from the group consisting of:

m is 1 to 4;

and z is 1 to 5.

87. The method of item 86, wherein m is equal to 1.

88. The method according to any one of claims 75 to 77, wherein the compound of formula (VIII) is the following compound:

where n is an integer from 4 to 16, preferably n is equal to 9 or 10.

89. The process according to any one of items 76 and 78 to 88, wherein the conversion in step (2) is a radical reaction using AIBN (azobisisobutyronitrile) as a radical initiator.

90. The method of any one of claims 76 and 78 to 89, wherein X is a bromine atom.

91. The method of any one of claims 76 and 78 to 90, wherein the converting in step (2) uses N-bromosuccinimide (NBS) as a reagent.

92. The process according to any one of claims 76 and 78 to 91, wherein the conversion in step (3) uses TBME (methyl tert-butyl ether) as solvent.

93. The process according to any one of claims 76 and 78 to 92, wherein the conversion in step (3) is carried out for at least 2 hours, preferably at ambient temperature.

94. The process according to any one of claims 76 and 78 to 93, wherein the conversion in step (3) is carried out for not more than 5 hours, preferably at ambient temperature.

95. The process according to any one of claims 76 and 78 to 94, wherein the conversion in step (3) is carried out for 3 hours, preferably at ambient temperature.

96. The process according to any one of claims 75 to 95, wherein the process is carried out on a scale to produce at least 100g, at least 1kg, at least 10kg, at least 100kg or at least 1000kg of the compound of formula (VIII).

Drawings

FIG. 1: virus inactivation efficiency of low concentrations of Triton X-100Reduced or Triton N-101Reduced when S/D treated IVIG-containing fluids at 17 ℃. + -. 1 ℃. Using a three component mixture, final concentrations of 0.04% to 0.06% Triton X-100Reduced or Triton N-101Reduced, 0.01% to 0.02% polysorbate 80, 0.01% to 0.02% TnBP were obtained (side by side comparison to the same concentration of Triton X-100). For two experiments with HIV (a and B, respectively), the time-dependent inactivation of the virus was expressed as a Reduction Factor (RF). Virus inactivation by S/D treatment with Triton X-100Reduced ("TX-100 Red.") or Triton N-101Reduced ("TN-101 Red.") was compared to that with Triton X-100 ("TX-100").

FIG. 2: virus inactivation efficiency of low concentrations of Triton X-100Reduced or Triton N-101Reduced when S/D treated IVIG-containing fluids at 17 ℃. + -. 1 ℃. Using a three component mixture, final concentrations of 0.04% to 0.06% Triton X-100Reduced or Triton N-101Reduced, 0.01% to 0.02% polysorbate 80, 0.01% to 0.02% TnBP were obtained (side by side comparison to the same concentration of Triton X-100). For two experiments using PRV (a and B, respectively), the time-varying inactivation of the virus was expressed as a virus Reduction Factor (RF). Virus inactivation by S/D treatment with Triton X-100Reduced ("TX-100 Red.") or Triton N-101Reduced ("TN-101 Red.") was compared to that with Triton X-100 ("TX-100").

FIG. 3: the virus inactivation efficiency of low concentrations of Brij C10 when the IVIG-containing fluids were subjected to S/D treatment at 17 ℃. + -. 1 ℃. Using a three component mixture, final concentrations of 0.04% to 0.06% Brij C10, 0.01% to 0.02% polysorbate 80, 0.01% to 0.02% TnBP were obtained (side by side comparison with Triton X-100 at the same concentration). For two experiments with HIV (a and B, respectively), the time-dependent inactivation of the virus was expressed as a virus Reduction Factor (RF). Viral inactivation by S/D treatment with Brij C10 was compared to viral inactivation by S/D treatment with Triton X-100 ("TX-100").

FIG. 4: the virus inactivation efficiency of low concentrations of Brij C10 when the IVIG-containing fluids were subjected to S/D treatment at 17 ℃. + -. 1 ℃. Using a three component mixture, final concentrations of 0.04% to 0.06% Brij C10, 0.01% to 0.02% polysorbate 80, 0.01% to 0.02% TnBP were obtained (side by side comparison with Triton X-100 at the same concentration). For two experiments using PRV (a and B, respectively), the time-varying inactivation of the virus was expressed as a virus Reduction Factor (RF). Viral inactivation by S/D treatment with Brij C10 was compared to viral inactivation by S/D treatment with Triton X-100 ("TX-100").

FIG. 5: the virus inactivation efficiency of low concentrations of Brij C10 when the IVIG-containing fluids were subjected to S/D treatment at 17 ℃. + -. 1 ℃. Using a three component mixture, final concentrations of 0.04% to 0.06% Brij C10, 0.01% to 0.02% polysorbate 80, 0.01% to 0.02% TnBP were obtained (side by side comparison with Triton X-100 at the same concentration). For two experiments with BVDV (a and B, respectively), the time-varying inactivation of the virus was expressed as the virus Reduction Factor (RF). Viral inactivation by S/D treatment with Brij C10 was compared to viral inactivation by S/D treatment with Triton X-100 ("TX-100").

FIG. 6 virus inactivation efficiency of low concentrations of Brij C10 when S/D treatment was performed on a liquid containing Human Serum Albumin (HSA) at 1 ℃. + -. 1 ℃Usinga three component mixture, resulting in final concentrations of 0.08% to 0.1% BrijC10, 0.02% to 0.03% polysorbate 80, 0.02% to 0.03% TnBP (side by side comparison to the same concentration of Triton X-100). for two experiments using X-Mu L V (A and B, respectively), the virus inactivation as a function of time was expressed in terms of the virus Reduction Factor (RF). Virus inactivation by S/D treatment using Brij C10 was compared to that by S/D treatment using Triton X-100 ("TX-100").

FIG. 7: the virus inactivation efficiency of low concentration of Brij C10 was found when HSA-containing fluids were subjected to S/D treatment at 1 ℃. + -. 1 ℃. Using a three component mixture, final concentrations of 0.08% to 0.1% Brij C10, 0.02% to 0.03% polysorbate 80, 0.02% to 0.03% TnBP were obtained (compared to the current data for Triton X-100). For two experiments with BVDV (a and B, respectively), the time-varying inactivation of the virus was expressed as the virus Reduction Factor (RF). Viral inactivation by S/D treatment with Brij C10 was compared to viral inactivation by S/D treatment with Triton X-100 ("TX-100").

FIG. 8 shows the virus inactivation efficiency of low concentrations of Brij C10 when S/D treatment was performed on HSA-containing fluids at 19 ℃. + -. 1 ℃. Using a three-component mixture, a final concentration of 0.08% to 0.1% Brij C10, 0.02% to 0.03% polysorbate 80, 0.02% to 0.03% TnBP was obtained (side-by-side comparison with the same concentration of Triton X-100). for two experiments with X-Mu L V (A and B, respectively), the time-dependent virus inactivation was expressed as a virus Reduction Factor (RF). Virus inactivation by S/D treatment with Brij C10 was compared to S/D treatment with Triton X-100 ("TX-100").

FIG. 9: low concentration of 4-tert-octylbenzyl alcohol polyethoxylate at 17 ℃. + -. 1 ℃ in S/D treatment of IVIG-containing fluids. Using a three component mixture, final concentrations of 0.04% to 0.06% 4-tert-octylbenzyl alcohol polyethoxylate, 0.01% to 0.02% polysorbate 80, 0.01% to 0.02% TnBP were obtained (side by side comparison with the same concentration of Triton X-100). For two experiments using PRV (a and B, respectively), the time-varying inactivation of the virus was expressed as a virus Reduction Factor (RF). Viral inactivation by S/D treatment with 4-tert-octylbenzyl alcohol polyethoxylate was compared to viral inactivation by S/D treatment with Triton X-100 ("TX-100"). Note that in (a), both detergents showed the same kinetics of inactivation. Solid symbols show values with residual infectivity (remainingness), non-solid symbols show a reduction factor below the detection limit.

FIG. 10 efficiency of viral inactivation of low concentrations of 4-tert-octylbenzyl alcohol polyethoxylate when S/D treated with Human Serum Albumin (HSA) at 1 ℃. + -. 1 ℃. Using a three-component mixture, a final concentration of 0.08% to 0.1% 4-tert-octylbenzyl alcohol polyethoxylate, 0.02% to 0.03% polysorbate 80, 0.02% to 0.03% TnBP (side by side comparison with the same concentration of Triton X-100) was obtained.

FIG. 11 Virus inactivation efficiency of low concentrations of 4-tert-octylbenzyl alcohol polyethoxylate when S/D treated with HAS in fluids at 19 ℃. + -. 1 ℃. Using a three component mixture, a final concentration of 0.08% to 0.1% 4-tert-octylbenzyl alcohol polyethoxylate, 0.02% to 0.03% polysorbate 80, 0.02% to 0.03% TnBP (side by side comparison to the same concentration of Triton X-100.) for two experiments with X-Mu L V (A and B, respectively), viral inactivation as a function of time is expressed in terms of the viral Reduction Factor (RF). Virus inactivation of S/D treated with 4-tert-octylbenzyl alcohol polyethoxylate is compared to S/D treated with TriX-100 ("TX-100").

FIG. 12: (A) low concentrations of 4-tert-octylbenzyl alcohol polyethoxylate or Triton X-100Reduced or Brij C10 were found to have virus inactivation efficiencies when the buffers containing HSA were detergent treated at 14 ℃. + -. 1 ℃. Using a single detergent treatment, a final concentration of 0.09% to 0.11% of 4-tert-octylbenzyl alcohol polyethoxylate or Triton X-100Reduced or Brij C10 (side-by-side comparison to the same concentration of Triton X-100) was obtained. For two experiments using BVDV, time-dependent viral inactivation was expressed as the mean viral Reduction Factor (RF). Viral inactivation by single detergent treatment with 4-tert-octylbenzyl alcohol polyethoxylate or Triton X-100Reduced ("TX-100 red.") or Brij C10 was compared to viral inactivation by single detergent treatment with Triton X-100 ("TX-100"). Note that 4-tert-octyl benzyl alcohol polyethoxylate and Triton X-100Reduced have the same inactivation kinetics as Triton X-100. Filled symbols show values with residual infectivity, non-filled symbols show a reduction factor below the detection limit. (B) 4-t-octyl benzyl alcohol polyethoxylate or Triton X-100Reduced virus inactivation efficiency when detergent treated in buffers containing HSA at 14 ℃. + -. 1 ℃. Using a single detergent treatment, a final concentration of 0.02% to 0.04% 4-tert-octylbenzyl alcohol polyethoxylate or Triton X-100Reduced (side-by-side comparison to the same concentration of Triton X-100) was obtained. For two experiments using BVDV, time-dependent viral inactivation was expressed as the mean viral Reduction Factor (RF). Virus inactivation by single detergent treatment with 4-tert-octylbenzyl alcohol polyethoxylate or Triton X-100Reduced ("TX-100 red.") was compared to that by single detergent treatment with Triton X-100 ("TX-100"). Filled symbols show values with residual infectivity, non-filled symbols show a reduction factor below the detection limit.

FIG. 13: (A) low concentrations of 4-tert-octylbenzyl alcohol polyethoxylate or Triton X-100Reduced virus inactivation efficiency when detergent treated in IVIG-containing fluids at 17 ℃. + -. 1 ℃. Using a single detergent treatment, a final concentration of 0.09% to 0.11% of 4-tert-octylbenzyl alcohol polyethoxylate or Triton X-100Reduced (side-by-side comparison to the same concentration of Triton X-100) was obtained. For two experiments using BVDV, time-dependent viral inactivation was expressed as the mean viral Reduction Factor (RF). Virus inactivation by single detergent treatment with 4-tert-octylbenzyl alcohol polyethoxylate or Triton X-100Reduced ("TX-100 red.") was compared to that by single detergent treatment with Triton X-100 ("TX-100"). Filled symbols show values with residual infectivity, non-filled symbols show a reduction factor below the detection limit. (B) Low concentrations of 4-tert-octylbenzyl alcohol polyethoxylate or Triton X-100Reduced virus inactivation efficiency when detergent treated in IVIG-containing fluids at 17 ℃. + -. 1 ℃. Using a single detergent treatment, a final concentration of 0.02% to 0.04% 4-tert-octylbenzyl alcohol polyethoxylate or Triton X-100Reduced (side-by-side comparison to the same concentration of Triton X-100) was obtained. For two experiments using BVDV, time-dependent viral inactivation was expressed as the mean viral Reduction Factor (RF). Virus inactivation by single detergent treatment with 4-tert-octylbenzyl alcohol polyethoxylate or Triton X-100Reduced ("TX-100 red.") was compared to that by single detergent treatment with Triton X-100 ("TX-100"). Filled symbols show values with residual infectivity, non-filled symbols show a reduction factor below the detection limit.

FIG. 14: low concentrations of 4-tert-octylbenzyl alcohol polyethoxylate or Triton X-100Reduced virus inactivation efficiency when detergent treated with factor VIII-containing fluids at 23 ℃. + -. 1 ℃. Using a single detergent treatment, a final concentration of 0.09% to 0.11% of 4-tert-octylbenzyl alcohol polyethoxylate or Triton X-100Reduced (side-by-side comparison to the same concentration of Triton X-100) was obtained. For two experiments using BVDV, time-dependent viral inactivation was expressed as the mean viral Reduction Factor (RF). Virus inactivation by single detergent treatment with 4-tert-octylbenzyl alcohol polyethoxylate or Triton X-100Reduced ("TX-100 red.") was compared to that by single detergent treatment with Triton X-100 ("TX-100"). Filled symbols show values with residual infectivity, non-filled symbols show a reduction factor below the detection limit.

Detailed Description

Unless defined otherwise below, terms used in the present invention should be understood according to the general meaning known to those skilled in the art.

All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes.

Definition of

The terms "virus having a lipid envelope", "lipid enveloped virus" and "enveloped virus" are used interchangeably herein and have the meaning known to the skilled person, e.g.the lipid enveloped viruses may be of the Herpesviridae family, e.g.pseudorabies virus (PRV), herpes simplex virus, varicella zoster virus, cytomegalovirus or Epstein-Barr virus (Epstein-Barr virus), the Hepadnaviridae family (Hepadnaviridae), e.g.hepatitis B virus, the Togaviridae family, e.g.Sindbis virus (Sindbis virus), rubella virus or alphavirus, the sandiviridae family (Arenaviridae), e.g.lymphocytic choriomeningitis virus (lmphocytic choromomeringitis virus), the Flaviviridae family, e.g.West Nile virus (Wenye virus), bovine viral diarrhea virus (BVDV virus, bovine viral dirreriviral dirreriviridae virus), Burkitt virus (Burkhoviridae virus), Buguvirus (Bureavirus), e.g.g.HIV), or Rhabdoviridae virus (Rhabdoviridae virus), e.g.g.HIV virus (Rhabdoviridae virus), or Rhabdoviridae virus (Rhabdoviridae), such as HIV), or Rhabdoviridae virus (Rhabdoviridae virus), or HIV-virus (Rhabdoviridae), such as HIV), or HIV-virus (Rhabdoviridae virus (HIV), or HIV-virus (Rhabdoviridae virus (Rhabdovirina virus (HIV), or HIV-virus (Rhabdovirinovirina virus (Rhabdovirina virus (HIV), or HIV-virus (Rhabdovirinovirinovirina virus (Rhabdovirinae virus (e), or HIV-virus (Rhabdovirinvirus (e virus (HIV), or HIV-virus (Rhabdovirinvirus (e virus (e.g.S virus (HIV), or HIV-virus (HIV), or HIV-virus (Rhabdovirinvirus (or HIV), or HIV-virus (e virus (Rhabdovirinvirus (HIV), or HIV-virus (Rhabdovirinvirus (or HIV), or HIV-virus (Rhabdovirinvirus (or HIV), or HIV (HIV), or.

As used herein, the term "inactivating a virus having a lipid envelope" refers to the ability to destroy a lipid enveloped virus infected cell. As will be appreciated by those skilled in the art, the ability of a lipoenveloped virus to infect cells (i.e., infectivity of a lipoenveloped virus) is typically assessed by determining the number of infectious viral particles in the fluid. Thus, as used herein, the term "inactivating a virus having a lipid envelope" or "inactivating a lipid enveloped virus" refers to reducing the number of infectious viral particles in solution.

Herein, the term "L og10 reduction value" or "L RV" is used interchangeably with the terms "viral reduction coefficient", "RF" or "R" in one embodiment, "L0 og10 reduction value" or "L1 RV" may be used as a measure of reduction of infectious viral particles in a liquid as used herein, "L og10 reduction value" or "L RV" is defined as the logarithm (base 10) of the ratio of infectious viral particles before viral inactivation to infectious viral particles after viral inactivation L RV value is specific for a given type of virus as is apparent to those skilled in the art that any L og10 reduction value (L RV) above 0 helps to improve the safety of methods and processes (e.g., biopharmaceutical production methods and processes.) the L og10 reduction value (L RV) achieved by the present methods is determined as known to those skilled in the art, e.g., the RV number of infectious particles in a liquid before and after viral inactivation of the present methods can be determined L.

The skilled person will be aware of various methods for measuring infectious viral particles in a liquid. For example, but not limited to, the concentration of infectious viral particles in a liquid can preferably be determined by plaque assay or by TCID₅₀Determined by an assay, more preferably by TCID₅₀Measured by an assay method. As used herein, "TCID₅₀Assay "refers to tissue culture infection dosimetry. TCID₅₀The assay is an end-point dilution assay, wherein TCID₅₀Values represent the concentration of virus necessary to induce cell death or pathological changes in 50% of the inoculated cell cultures.

As used herein, the term "surfactant" refers to a compound that reduces the surface tension between two liquids or between a liquid and a solid. Surfactants may be used as soil release agents, wetting agents, emulsifiers, foaming agents and dispersing agents.

With respect to the present invention, terms (e.g., "adding to," "adding to," or "adding") with respect to first-mentioned and last-mentioned solvents, detergents, and/or liquids include the case where the first-mentioned solvent, detergent, and/or liquid is added to the last-mentioned solvent, detergent, and/or liquid. However, these terms are also intended to include the case where a later-mentioned solvent, detergent, and/or liquid is added to a first-mentioned solvent, detergent, and/or liquid. Thus, terms such as "added to," "added to," or "added to" are not intended to indicate whether a first-mentioned solvent, detergent, and/or liquid is added to a later-mentioned solvent, detergent, and/or liquid, and vice versa.

As known to those skilled in the art, the term "detergent" is used according to its general meaning known in the art and specifically includes surfactants that can permeabilize lipid membranes. For example, it has been proposed to solubilize amphiphilic membrane proteins by binding to the hydrophobic part of the protein with the detergent Triton X-100 and deoxycholate (see Simons et al, 1973). Detergents are classified into three broad categories based on their charge. Anionic detergents contain anionic (i.e., negatively charged) hydrophilic groups. Exemplary anionic detergents are: tetradecyltrimethylammonium bromide, dodecyltrimethylammonium bromide, sodium lauryl sulfate, Sodium Dodecyl Sulfate (SDS), cetrimide (cetrimide), and hexadecyltrimethylammonium bromide. Cationic detergents contain cationic (i.e., positively charged) hydrophilic groups. Exemplary cationic detergents are: benzalkonium chloride, cetyltrimethylammonium bromide (CTAB), cetylpyridinium chloride (CPC) and benzethonium chloride (BZT). As used herein, the term "nonionic detergent" refers to a detergent that does not have a positive or negative charge, exemplary nonionic detergents are: sorbitan esters (sorbitan esters) (sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan tristearate, sorbitan monooleate, sorbitan trioleate), polysorbates (polyoxyethylene (20) sorbitan monolaurate (polysorbate 20), polyoxyethylene (20) sorbitan monopalmitate, polyoxyethylene (20) sorbitan monostearate, polyoxyethylene (20) sorbitan tristearate, polyoxyethylene (20) sorbitan trioleate, polyoxyethylene (20) sorbitan monooleate (Tween 80/polysorbate 80)), poloxamers (poloxamer 407, poloxamer 188) and cremophor (cremophor). Detergents of the invention are described in the preferred embodiments, including but not limited to Triton N-101Reduced, Triton X-100Reduced, and Brij C10.

As used herein, the term "non-phenolic" is used interchangeably with the term "phenolic-free". Non-phenolic detergents as used herein refers to detergents that do not contain any phenolic functionality. The term "aroma" has the meaning known to those skilled in the art. Non-aromatic detergents as used in the present invention refer to detergents which do not contain any aromatic rings.

As used herein, the term "environmentally compatible" has the meaning known to those skilled in the art. In a preferred embodiment of the invention, with respect to detergents, the term "environmentally compatible" means that the detergent does not act as an endocrine disrupter. Endocrine disruptors are exogenous substances that alter the functioning of the endocrine system and thus can cause adverse health effects in the intact organism or its offspring or (sub) population. The skilled person will be aware of various methods of identifying endocrine disruptors. More information about endocrine disruptors and their assessment, for example, can be found in "ECHA Support documents for identification of4- (1,1,3,3-tetramethylbutyl) phenols, ethoxylated assortments of top high connected phenols, product to the third identification to the top high connected phenols (4- (1,1,3,3-tetramethylbutyl) phenols), the latter of these, the latter of the high ethoxylated products to the third identification, the latter of the third identification to the third identification of the level of the first mentioned compounds, and PBTs/vPvBSs (ECHA Support documents for identifying highly ethoxylated substances of interest, 4- (1,1,3,3-tetramethylbutyl) phenols with highly degraded properties of these, 1, 3-tetramethylbutyl) phenols with high degree of degradation of these compounds, they may have a severe impact on the environment, causing the same degree of concern as CMR and PBT/vPvB) ", which is passed on 12/2012, the entire contents of which are incorporated herein by reference in their entirety and for all purposes; or the manual "Global Association of the Sate-of-the-Science of Endocrine disorders" (WHO/PCS/EDC/02.2), published by the World Health organization International chemical Safety program of the World Health organization, the entire contents of which are incorporated herein by reference in their entirety and for all purposes.

As used herein, the term "solvent" has the meaning known to those skilled in the art. In a preferred embodiment of the present invention, an organic solvent is used in the process of the present invention. Particularly useful organic solvents create an environment that promotes contact between the detergent and the lipoprotein envelope of the lipid-enveloped virus. As such, organic solvents that facilitate such contact are preferably used in the process of the present invention, including but not limited to ethers, alcohols, alkyl phosphates (e.g., dialkyl or trialkyl phosphates), or any combination thereof.

Ether solvents useful in the processes disclosed herein include those of the formula R1-O-R2 wherein R1 and R2 are independently C1-C18 alkyl or C1-C18 alkenyl groups, preferably C1-C18 alkyl or C1-C18 alkenyl groups, which may contain an oxygen atom or a sulfur atom. Non-limiting examples of ethers include: dimethyl ether, diethyl ether, ethyl propyl ether, methyl butyl ether, methyl isopropyl ether, and methyl isobutyl ether. Alcohol solvents useful in the processes disclosed herein include those having a C1-C8 alkyl group or a C1-C8 alkenyl group. Non-limiting examples of alcohols include: methanol, ethanol, propanol, isopropanol, n-butanol, isobutanol, n-pentanol and isopentanol. Alkyl phosphate ester solvents useful in the processes disclosed herein include those having a C1-C18 alkyl group or a C1-C18 alkenyl group, the C1-C18 alkyl group or the C1-C18 alkenyl group may contain an oxygen atom or a sulfur atom. Non-limiting examples of alkyl phosphates include: dialkyl phosphates such as di (n-butyl) phosphate, di (tert-butyl) phosphate, di (n-hexyl) phosphate, di (2-ethylhexyl) phosphate, di (n-decyl) phosphate or di (n-butyl) phosphate; and trialkyl phosphates such as tri (n-butyl) phosphate, tri (t-butyl) phosphate, tri (n-hexyl) phosphate, tri (2-ethylhexyl) phosphate or tri (n-decyl) phosphate.

As used herein, the term "biomedical product" is known in the art and refers to a product whose active substance is a biological substance (e.g., a biological substance produced by a mammalian cell or microorganism). As used herein, the biopharmaceutical product used in the method of the present invention is not limited to a final manufactured product, but preferably also includes intermediate products at any stage of the manufacturing process.

As used herein, the term "biopharmaceutical" has the meaning known to those skilled in the art. Biopharmaceuticals include recombinant biopharmaceuticals and biopharmaceuticals of other origin, such as those obtained from human plasma.

As used herein, the term "depth filter" has the meaning known in the art. In particular, such filters (e.g., gradient-density depth filters) enable filtration within the depth of the filter material. One common class of such filters are those that include a random matrix of bonded (or otherwise fixed) fibers to form a labyrinth of tortuous flow paths. Particle separation in these filters is typically caused by entrapment or adsorption by the fibrous matrix. The most common depth filtration media used for bioprocessing of cell culture fluids and other feedstocks are cellulose fibers, filter aids (e.g., DE), and positively charged resin binders. Unlike absolute filters, depth filtration media retains particles throughout the porous media, allowing retention of particles larger and smaller than the pore size. Particle retention is believed to involve size exclusion and adsorption through hydrophobic, ionic and other interactions.

As used herein, the term "purified biopharmaceutical" has a meaning known to those skilled in the art and refers to the separation of a biopharmaceutical from other substances that may be contained in a mixture of the present invention. In a preferred embodiment of the invention, the term "purifying the biopharmaceutical" refers to separating the biopharmaceutical from the detergent of the invention.

The term "chromatography" is used according to the meaning known in the art. It includes any chromatographic technique that separates the target analyte (e.g., a target molecule, such as a biological drug) from other molecules present in the mixture. In general, analytes of interest will be separated from other molecules due to the different rates of migration of individual molecules in a mixture through an immobilization medium under the influence of a mobile phase or binding process and an elution process.

The terms "chromatography resin" and "chromatography medium" are used interchangeably herein and refer to any kind of phase (e.g., solid phase) that separates an analyte of interest (e.g., a target molecule, such as a biopharmaceutical) from other molecules present in a mixture. In general, analytes of interest will be separated from other molecules due to the different rates of migration of individual molecules in a mixture through a stationary solid phase under the influence of a mobile phase or a binding process and an elution process. Examples of various types of chromatographic media include: for example, cation exchange resins, cation exchange membranes, affinity resins, anion exchange membranes, hydrophobic interaction resins, and ion exchange monolithic columns (monolith).

As used herein, the term "pharmaceutical formulation" has the meaning known to those skilled in the art and refers to any formulation suitable for administration to a patient. Pharmaceutical formulations may be prepared according to methods known in the art. For example, for any biopharmaceutical present in a formulation, the skilled person will be able to select and add preferred further ingredients, including buffers, stabilizers, surfactants, antioxidants, chelating agents, and/or preservatives and the like.

As used herein, the term "solvent/detergent mixture" has the meaning known to those skilled in the art. In a preferred embodiment, the solvent/detergent mixture used in the present invention comprises at least one solvent other than water and at least one detergent. The solvent used in the present invention is preferably an organic solvent, and most preferably tri-n-butyl phosphate. The number of different solvents and/or detergents contained in the mixture is not particularly limited. For example, according to the present invention, a solvent/detergent mixture may consist of tri-n-butyl phosphate, polysorbate 80 and a polyoxyethylene ether detergent.

It is understood that when used in the present disclosure to mean a range of values, the term "between" includes the indicated lower and upper limits of the respective range. For example, when the indicated temperature is 0 ℃ to 10 ℃, this includes temperatures of 0 ℃ and 10 ℃. Similarly, when the variable x is represented as an integer from 4 to 16, for example, it includes integers 4 and 16.

It is to be understood that the term "1 hour" as used herein is not limited to the precise 60 minutes. As used herein, the term "1 hour" is understood to relate to 60 minutes ± 5 minutes, preferably 60 minutes ± 2 minutes.