METHODS AND KITS FOR PREPARING PATHOGEN-INACTIVATED WHOLE BLOOD
Provided are methods for preparing pathogen-inactivated whole blood and other pathogen-inactivated blood product compositions, as well as kits and compositions related thereto.
1 . A method of preparing a pathogen inactivated whole blood composition, comprising:
(a) mixing in a first container
(i) a whole blood composition;
(ii) an effective amount of a pathogen inactivation compound (PIC); and
optionally (iii) an effective amount of a quencher;
(b) incubating in the first container the mixture of the whole blood composition, the PIC and optionally the quencher, under conditions and for a time sufficient to inactivate a pathogen, if present;
(c) transferring the mixture under sterile conditions to a second container; and
(d) incubating in the second container the mixture under conditions and for a time sufficient such that at least one of (i)-(iii) applies:
(i) the concentration of the PIC after incubating in the second container is at least 25% less than the concentration of the PIC in the mixture at the time of transfer step (c);
(ii) the concentration of the PIC after incubating in the second container is at least 10 5 fold less than the concentration of the PIC in the mixture of step (a); or
(iii) the concentration of the PIC after incubating in the second container is about 1 nm or less,
thereby yielding the pathogen inactivated whole blood composition.
2 . The method of claim 1 , wherein the concentration of the PIC in the mixture of step (a) is at least about 0.1 mM.
3 . The method of claim 1 or claim 2 , wherein the concentration of the PIC in the mixture of step (a) is about 0.1 mM to about 1.0 mM.
4 . The method of any one of claims 1 - 3 , wherein the concentration of the PIC in the mixture of step (a) is about 0.2 mM.
5 . The method of any one of claims 1 - 4 , wherein the concentration of the PIC after step (d) is less than about 2 nM.
6 . The method of any one of claims 1 - 5 , wherein the concentration of the PIC after step (d) is less than about 1 nM.
7 . The method of any one of claims 1 - 6 , wherein the PIC comprises a functional group which is, or which forms, a reactive electrophilic group.
8 . The method of claim 7 , wherein the functional group is selected from the group consisting of a mustard, a mustard intermediate, and a mustard equivalent.
9 . The method of claim 7 or claim 8 , wherein the functional group is, or is capable of forming, an aziridinium ion.
10 . The method of any one of claims 1 - 9 , wherein the reactive electrophilic group is capable of reacting with nucleic acids.
11 . The method of any one of claims 1 - 10 , wherein the PIC comprises a nucleic acid binding ligand.
12 . The method of claim 11 , wherein the nucleic acid binding ligand is an intercalator.
13 . The method of claim 12 , wherein the intercalator is an acridine.
14 . The method of any one of claims 11 - 13 , wherein the PIC comprises a frangible linker linking the functional group and the nucleic acid binding ligand.
15 . The method of any one of claims 1 - 14 , wherein the PIC is β-alanine, N-(acridin-9-yl), 2-[bis(2-chloroethyl)amino]ethyl ester.
16 . The method of any one of claims 1 - 15 , wherein step (a) comprises mixing in the first container a whole blood composition, an effective amount of a PIC, and an effective amount of a quencher.
17 . The method of claim 16 , wherein the concentration of the quencher in the mixture of step (a) is at least about 1 mM.
18 . The method of claim 16 or claim 17 , wherein the concentration of the quencher in the mixture of step (a) is about 1 mM to about 10 mM.
19 . The method of any one of claims 16 - 18 , wherein the concentration of the quencher in the mixture of step (a) is about 2 mM.
20 . The method of any one of claims 16 - 19 , wherein the concentration of the PIC in the mixture of step (a) is about 0.2 mM and the concentration of the quencher in the mixture of step (a) is about 2 mM.
21 . The method of any one of claims 16 - 20 , wherein the quencher comprises a thiol group, wherein the thiol is capable of reacting with a reactive electrophilic group of the PIC.
22 . The method of any one of claims 16 - 21 , wherein the quencher comprises cysteine or a derivative of cysteine.
23 . The method of any one of claims 16 - 22 , wherein the quencher is glutathione or a pharmaceutically acceptable salt thereof.
24 . The method of any one of claims 16 - 23 , wherein the quencher is glutathione monosodium salt.
25 . The method of any one of claims 1 - 24 , wherein the mixture of step (a) further comprises an anticoagulant.
26 . The method of any one of claims 1 - 25 , wherein the whole blood composition further comprises an anticoagulant.
27 . The method of any one of claims 1 - 25 , wherein the first container contains an anticoagulant and the whole blood composition is mixed with the anticoagulant in the first container.
28 . The method of any one of claims 1 - 27 , wherein the whole blood composition has a volume of about 250 mL to about 700 mL.
29 . The method of any one of claim 25 - 28 , wherein the ratio of anticoagulant to whole blood in the whole blood composition is about 1:5 to about 1:9.
30 . The method of any one of claims 1 - 29 , wherein the whole blood composition is added into the first container through a first inlet of the first container.
31 . The method of any one of claims 1 - 30 , wherein the whole blood composition is added into the first container from a donor collection container containing the whole blood composition.
32 . The method of any one of claims 1 - 30 , wherein the whole blood composition is added into the first container by collecting the whole blood composition from a whole blood donor into the first container.
33 . The method of claim 32 , wherein the whole blood composition is collected from a whole blood donor into the first container.
34 . The method of any one of claims 1 - 33 , wherein the PIC and optionally the quencher are added into the first container through a first inlet of the first container.
35 . The method of any one of claims 1 - 33 , wherein the PIC and optionally the quencher are added into the first container through a second inlet of the first container.
36 . The method of claim 34 or claim 35 , wherein the PIC and optionally the quencher are added into the first container through a conduit extending between a container containing the PIC (PIC container) and optionally a container containing the quencher (quencher container) and an inlet of the first container, and providing a fluid communication path for transferring the PIC from the PIC container and optionally for transferring the quencher from the optional quencher container, into the first container.
37 . The method of claim 36 , wherein the PIC and the quencher are added into the first container through a conduit extending between a PIC container and a quencher container and an inlet of the first container, and wherein the conduit comprises at one end: (a) a first adaptor for coupling the PIC container to the conduit, (b) a second adaptor for coupling the quencher container to the conduit, and optionally (c) a displacement device suitable for increasing the transfer of the PIC and the quencher into the first container.
38 . The method of any one of claims 1 - 37 , wherein the incubating step (b) is about 2 hours to about 24 hours.
39 . The method of claim 38 , wherein the incubating step (b) is about 18 hours to about 22 hours.
40 . The method of any one of claims 1 - 39 , wherein the incubating step (b) is at room temperature.
41 . The method of any one of claims 1 - 40 , wherein the transferring step (c) is through a conduit extending between the first container and the second container.
42 . The method of claim 41 , wherein the conduit connects a first outlet of the first container to a first inlet of the second container.
43 . The method of any one of claims 1 - 42 , wherein the incubating step (d) is about 1 hour to about 8 hours.
44 . The method of claim 43 , wherein the incubating step (d) is about 2 hours to about 4 hours.
45 . The method of any one of claims 1 - 44 , wherein the incubating step (d) is at room temperature.
46 . The method of any one of claims 1 - 45 , further comprising transferring the pathogen-inactivated whole blood composition from the second container to a third container.
47 . The method of any one of claims 1 - 46 , further comprising (e) storing the pathogen inactivated whole blood composition at about 2-6° C.
48 . The method of any one of claims 1 - 47 , further comprising preparing from the pathogen-inactivated whole blood composition one or more of a red blood cell composition, a plasma composition, or a platelet composition.
49 . The method of any one of claims 1 - 48 , further comprising preparing from the pathogen-inactivated whole blood composition a plasma composition, and preparing a cryoprecipitate composition from the plasma composition.
50 . A pathogen-inactivated whole blood composition prepared by the method of any one of claims 1 - 49 .
51 . A red blood cell composition prepared by the method of claim 48 .
52 . A cryoprecipitate composition prepared by the method of claim 49 .
53 . A method of infusing a pathogen-inactivated whole blood composition into a subject in need thereof, the method comprising infusing into the subject a pathogen-inactivated whole blood composition prepared by the method of any one of claims 1 - 47 or a pathogen-inactivated whole blood composition of claim 50 .
54 . The method of claim 53 , wherein the method comprises coupling an infusion line assembly to the second container and infusing the pathogen-inactivated whole blood composition into the subject.
55 . The method of claim 53 , wherein the method comprises infusing the pathogen-inactivated whole blood composition through an infusion line assembly coupled to the second container.
56 . A kit for preparing a pathogen-inactivated whole blood composition, comprising:
(a) a vascular access device for drawing whole blood from a donor;
(b) a first container suitable for mixing the whole blood and a pathogen inactivation compound (PIC), wherein the first container comprises at least a first inlet and a first outlet;
(c) a first conduit extending between the vascular access device and the first container and providing a fluid communication path for transferring the whole blood from the donor into the first container; and
(d) a container containing a pathogen inactivation compound (PIC container).
57 . The kit of claim 56 , wherein the PIC container is configured to be coupled by a fluid communication path to the first container.
58 . The kit of claim 57 , wherein the PIC container is configured to be coupled to the first conduit.
59 . The kit of claim 56 , wherein the PIC container is coupled by a fluid communication path to the first container.
60 . The kit of claim 56 , wherein the first container further comprises a second inlet and the PIC container is configured to be coupled by a fluid communication path to the second inlet of the first container.
61 . The kit of claim 56 , wherein the first container further comprises a second inlet and the PIC container is coupled by a fluid communication path to the second inlet of the first container.
62 . The kit of any one of claims 56 - 61 , further comprising a container containing a quencher (quencher container).
63 . The kit of claim 62 , wherein the quencher container is configured to be coupled by a fluid communication path to the first container.
64 . The kit of claim 63 , wherein the quencher container is configured to be coupled to the first conduit.
65 . The kit of claim 62 , wherein the quencher container is coupled by a fluid communication path to the first container.
66 . The kit of claim 63 , wherein the quencher container is configured to be coupled by a fluid communication path to the second inlet of the first container.
67 . The kit of claim 65 , wherein the quencher container is coupled by a fluid communication path to the second inlet of the first container.
68 . The kit of any one of claims 62 - 67 , wherein the PIC container and the quencher container are the same container.
69 . The kit of any one of claims 56 - 68 , further comprising a second conduit wherein the second conduit comprises at one end at least one adaptor for coupling a PIC container to the second conduit.
70 . The kit of claim 69 , wherein the second conduit comprises at one end a first adaptor for coupling a PIC container to the second conduit and a second adaptor for coupling a quencher container to the second conduit.
71 . The kit of any one of claims 56 - 68 , further comprising a second conduit wherein the second conduit is coupled at one end to a PIC container and/or a quencher container.
72 . The kit of any one of claims 69 - 71 , wherein the second conduit is configured to be coupled to the first container.
73 . The kit of any one of claims 69 - 71 , wherein the second conduit is coupled to the first container.
74 . The kit of any one of claims 69 - 71 , wherein the second conduit is configured to be coupled to the first conduit, providing a fluid communication path between the PIC container and/or quencher container and the first container.
75 . The kit of any one of claims 56 - 74 , further comprising a displacement device suitable for increasing the transfer of the PIC and/or the quencher into the first container.
76 . The kit of any one of claims 56 - 75 , wherein the first container contains an anticoagulant.
77 . The kit of any one of claims 56 - 76 , further comprising a second container suitable for storing a pathogen-inactivated whole blood composition, wherein the second container comprises at least a first inlet and a first outlet.
78 . The kit of claim 77 , further comprising a third conduit extending between the first container and the second container and providing a fluid communication path for transferring a mixture comprising a whole blood composition and a PIC from the first container into the second container.
79 . The kit of claim 77 or claim 78 , wherein the kit comprises a third container suitable for storing a pathogen-inactivated whole blood composition, wherein the third container comprises at least a first inlet and a first outlet, and wherein the kit further comprises a fourth conduit extending between the second container and the third container and providing a fluid communication path for transferring a mixture comprising a whole blood composition and a PIC from the second container into the third container.
80 . The kit of any one of claims 56 - 79 , further comprising an infusion line assembly for infusing a pathogen-inactivated whole blood composition into a subject.
81 . The kit of claim 77 or claim 78 , further comprising an infusion line assembly for infusing a pathogen-inactivated whole blood composition into a subject, wherein the infusion line assembly is coupled to the second container.
82 . The kit of claim 81 , wherein the infusion line assembly comprises a second vascular access device and an infusion line conduit extending between the first outlet of the second container and the second vascular access device, wherein the infusion line conduit provides a fluid communication path for sterilely transferring the pathogen-inactivated whole blood composition from the second container to the vascular access device.
83 . The kit of any one of claims 56 - 82 , wherein the PIC comprises a functional group which is, or which forms, a reactive electrophilic group.
84 . The kit of any one of claims 56 - 83 , wherein the PIC comprises a nucleic acid binding ligand.
85 . The kit of any one of claims 56 - 84 , wherein the PIC is β-alanine, N-(acridin-9-yl), 2-[bis(2-chloroethyl)amino]ethyl ester.
86 . The kit of any one of claims 62 - 85 , wherein the quencher comprises a thiol group, wherein the thiol is capable of reacting with a reactive electrophilic group of the PIC.
87 . The kit of any one of claims 62 - 86 , wherein the quencher is glutathione or a pharmaceutically acceptable salt thereof.
88 . A kit for preparing a pathogen-inactivated whole blood composition, comprising:
(a) a first container suitable for mixing a whole blood composition, a pathogen inactivation compound (PIC) and a quencher, wherein the first container comprises at least a first inlet and a first outlet;
(b) a second container suitable for storing a pathogen-inactivated whole blood composition, wherein the second container comprises at least a first inlet and a first outlet;
(c) a container containing a pathogen inactivation compound (PIC container);
(d) a container containing a quencher (quencher container);
(e) a first conduit, wherein the first conduit at one end is coupled to or configured to be coupled to a PIC container and a quencher container, and wherein the first conduit further comprises a displacement device suitable for increasing the transfer of the PIC and the quencher into the first container; and
(f) a second conduit extending between the first container and the second container and providing a fluid communication path for transferring the whole blood composition from the first container to the second container.
89 . The kit of claim 88 , wherein the first conduit further comprises at least one adaptor for coupling the first conduit to a PIC container and a quencher container.
90 . The kit of claim 89 , wherein the first conduit comprises a first adaptor for coupling a PIC container to the first conduit and a second adaptor for coupling a quencher container to the first conduit.
91 . The kit of any one of claims 88 - 90 , wherein the first conduit is coupled to a PIC container and a quencher container.
92 . The kit of any one of claims 88 - 91 , wherein the PIC container and the quencher container are the same container.
93 . The kit of any one of claims 88 - 92 , wherein the first conduit is coupled or configured to be coupled to the first container, providing a fluid communication path between the PIC container and the quencher container and the first container.
94 . The kit of claim 93 , wherein the first conduit is configured to be coupled to an inlet of the first container.
95 . The kit of claim 93 , wherein the first conduit is coupled to an inlet of the first container.
96 . The kit of claim 88 - 95 , wherein the first container further comprises a second inlet, and wherein the first conduit is coupled or configured to be coupled to the second inlet of the first container.
97 . The kit of any one of claims 88 - 95 , further comprising a vascular access device for drawing whole blood from a donor and third conduit extending between the vascular access device and the first inlet of the first container and providing a fluid communication path for transferring the whole blood from the donor into the first container.
98 . The kit of claim 97 , wherein the first conduit is configured to be coupled to the third conduit, providing a fluid communication path between the PIC container and the quencher container and the first container.
99 . The kit of any one of claims 88 - 98 , wherein the first container contains an anticoagulant.
100 . The kit of any one of claims 88 - 99 , wherein the kit comprises a third container suitable for storing a pathogen-inactivated whole blood composition, wherein the third container comprises a first inlet and a first outlet, and wherein the kit further comprises a fourth conduit extending between the second container and the third container and providing a fluid communication path for transferring the whole blood composition from the second container into the third container.
101 . The kit of any one of claims 88 - 100 , further comprising an infusion line assembly for infusing a pathogen-inactivated whole blood composition into a subject.
102 . The kit of any one of claims 88 - 101 , further comprising an infusion line assembly for infusing a pathogen-inactivated whole blood composition into a subject, wherein the infusion line assembly is coupled to the second container.
103 . The kit of any one of claims 88 - 102 , further comprising an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject, wherein the infusion line assembly comprises a second vascular access device and an infusion line conduit extending between the first outlet of the second container and the second vascular access device, wherein the infusion line conduit provides a fluid communication path for sterilely transferring the pathogen-inactivated whole blood composition from the second container to the vascular access device.
104 . The kit of any one of claims 88 - 103 , wherein the PIC is comprises a functional group which is, or which forms, a reactive electrophilic group.
105 . The kit of any one of claims 88 - 104 , wherein the PIC comprises a nucleic acid binding ligand.
106 . The kit of any one of claims 88 - 105 , wherein the PIC is β-alanine, N-(acridin-9-yl), 2-[bis(2-chloroethyl)amino]ethyl ester.
107 . The kit of any one of claims 88 - 106 , wherein the quencher comprises a thiol group, wherein the thiol is capable of reacting with a reactive electrophilic group of the PIC.
108 . The kit of any one of claims 88 - 107 , wherein the quencher is glutathione or a pharmaceutically acceptable salt thereof.