US6841535B2
Expired / Lapsed
US
⏱️ 0.0 years remaining
Peptide-mediated transfection agents and methods of use
Assignee: Centre National de la Recherche Scientifique
•
Filing: 2001-07-26
•
Granted: 2005-01-11
•
Expires: 2021-07-26
Abstract
Abstract Peptides that are useful as transfection agents are described. The peptides can facilitate the efficient cellular internalization of a broad range and size of compounds that when non-covalently complexed therewith are efficiently internalized into a cell. Advantages include but are not limited to excellent transfection efficiency, relatively low toxicity, internalization by a broad host cell spectrum, and the simplicity and cost-effectiveness that arise from not having to covalently complex the peptide with a specific molecule to be delivered. Applications include but are not limited to the delivery of diagnostics and therapeutics, as well as drug discovery, gene discovery, and the analysis and/or manipulation of other cellular and molecular biological functions. Claims are made for transfection agents, compositions of matter, including pharmaceutical compositions, reagent kits, methods of delivery, and methods of identification of additional peptides for performing and/or including in the same.
Inventors
- 👤 Gilles Divita
- 👤 May C. Morris
- 👤 Jean Mery
- 👤 Frederic Heitz
- 👤 Joseph Fernandez
- 👤 John Archdeacon
- 👤 Kyle Horndorp
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 20:08
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/87; A; A61; A61K; A61K47/00; A61K47/50; A61K47/51; A61K47/62; A61K47/64; C07; C07K; C07K14/00; C07K14/001; C07K7/00; C07K7/04; C07K7/08; A61K38/00
Claims Analysis
Total Claims: 124
Claim 1
Independent Claim
187 words
A transfection agent comprising a peptide of between about 16 to 30 amino a acid residues, said peptide comprising: a) a hydrophobic domain comprising at least four amino acid residues, wherein the hydrophobic domain comprises a plurality of aromatic amino acid residues and a first hydrophobic locus spaced by at least one amino acid residue from a second hydrohpobic locus, wherein each of the first and second hydrophobic loci comprise one or more hydrophobic amino acid residues independently selected from the group consisting of Phe, Tyr, Trp, Thr, Met, Leu, Val, Ile, Ala, and His, and wherein at least two of said plurality of aromatic amino acid residues occur in a pair in the first hydrophobic locus or the second hydrophobic locus; b) a hydrophilic domain comprising up to about twelve amino acid residues, wherein the hydrophilic domain comprises a plurality of basic amino acid residues; c) optionally a spacer sequence between said hydrophobic and said hydrophilic domain, wherein said spacer comprises from between one to about ten amino acid residues; and d) further optionally a functional group conjugated to one or more termini of said peptide.
Claim 2
Dependent Claim
187 words
A transfection agent comprising a peptide of between about 16 to 30 amino a acid residues, said peptide comprising: a) a hydrophobic domain comprising at least four amino acid residues, wherein the hydrophobic domain comprises a plurality of aromatic amino acid residues and a first hydrophobic locus spaced by at least one amino acid residue from a second hydrohpobic locus, wherein each of the first and second hydrophobic loci comprise one or more hydrophobic amino acid residues independently selected from the group consisting of Phe, Tyr, Trp, Thr, Met, Leu, Val, Ile, Ala, and His, and wherein at least two of said plurality of aromatic amino acid residues occur in a pair in the first hydrophobic locus or the second hydrophobic locus; b) a hydrophilic domain comprising up to about twelve amino acid residues, wherein the hydrophilic domain comprises a plurality of basic amino acid residues; c) optionally a spacer sequence between said hydrophobic and said hydrophilic domain, wherein said spacer comprises from between one to about ten amino acid residues; and d) further optionally a functional group conjugated to one or more termini of said peptide.
Claim 3
Dependent Claim
28 words
A transfection agent according to claim 1 wherein said hydrophilic domain is a cation-rich sequence comprised of at least two lysine residues within a span of seven residues.
Claim 4
Dependent Claim
33 words
A transfection agent according to claim 1 wherein at least two of said plurality of aromatic amino acid residues occur in a pair in the first hydrophobic locus or the second hydrophobic locus.
Claim 5
Dependent Claim
23 words
A transfection agent according to claim 2 wherein two or more of said at least two lysine residues are adjacent to one another.
Claim 6
Dependent Claim
25 words
A transfection agent according to claim 1 wherein said plurality of aromatic amino acid residues is between three and five aromatic amino acid residues inclusive.
Claim 7
Dependent Claim
21 words
A transfection agent according to claim 1 wherein said plurality of aromatic amino acid residues comprises at least two tryptophan residues.
Claim 8
Dependent Claim
50 words
A transfection agent according to claim 1 comprising two pairs of aromatic amino acid residues, the first pair being disposed in the first hydrophobic locus and the second pair being disposed in the second hydrophobic locus, and wherein said first and second hydrophobic loci are separated by two amino acids.
Claim 9
Dependent Claim
24 words
A transfection agent according to claim 7 wherein said two amino acids separating the first and second hydrophobic loci consist of hydrophilic amino acids.
Claim 10
Dependent Claim
23 words
A transfection agent according to claim 8 wherein said hydrophilic amino acids separating the first and second hydrophobic loci are Glu and Thr.
Claim 11
Dependent Claim
14 words
A transfection agent according to claim 1 wherein said peptide is a synthetic peptide.
Claim 12
Dependent Claim
34 words
A transfection agent according to claim 1 wherein said optional spacer sequence comprises one or more amino acid residues selected from the group consisting of proline, glycine, tyrosine, serine, glutamine, and non-charged amino acids.
Claim 13
Dependent Claim
37 words
A transfection agent according to claim 1 wherein said hydrophobic domain comprises a motif (Trp/Tyr)-(Trp/Tyr)-Xaa-Xaa-(Trp/Tyr), wherein the first and second hydrophobic loci are spaced by two amino acids, Xaa-Xaa, wherein each Xaa is a hydrophilic amino acid.
Claim 14
Dependent Claim
41 words
A transfection agent according to claim 1 wherein said agent is covalently affixed to a compound selected from the group consisting of a nucleic acid, a peptide, a protein, an antibody, and a derivatives or analog of any of the foregoing.
Claim 15
Dependent Claim
41 words
A transfection agent according to claim 1 wherein said agent is non-covalently complexed with a compound selected from the group consisting of a nucleic acid, a peptide, a protein, an antibody, and a derivative or analog of any of the foregoing.
Claim 16
Dependent Claim
55 words
A transfection agent according to claim 1 that further comprises a functional group covalently attached to a terminus of said peptide, wherein the functional group is selected from the group consisting of a cystearnine group, a methyl group, and an alkyl group, and, with respect to the amino terminus of said peptide, an acyl group.
Claim 17
Dependent Claim
43 words
A transfection agent according to claim 1 wherein said hydrophilic domain comprises the sequence Lys-Arg-Lys, and wherein said agent further comprises a spacer sequence that comprises at least three amino acid residues of which at least one is a proline or glutamine residue.
Claim 18
Dependent Claim
38 words
A transfection agent according to claim 1 that further comprises a functional group to which is covalently conjugated a molecule selected from the group consisting of a stabilizer, a coupler, a dye, a ligand, and an enzymatic substrate.
Claim 19
Dependent Claim
46 words
A commercial transfection kit comprising at least one transfection agent according to claim 1 in either aqueous or lyophilized form, said kit further comprising one or more components selected from the group consisting of buffers, positive controls, cells to be transfected, phospholipids, and instructions for use.
Claim 20
Dependent Claim
12 words
A pharmaceutical composition comprising a transfection agent according to claim 1 .
Claim 21
Dependent Claim
23 words
A pharmaceutical composition according to claim 19 wherein said transfection agent is non-covalently complexed with a compound to be delivered to a cell.
Claim 22
Dependent Claim
26 words
A pharmaceutical composition according to claim 20 wherein said compound comprises a member selected from the group consisting of a diagnostic compound and a therapeutic compound.
Claim 23
Dependent Claim
33 words
A pharmaceutical composition according to claim 20 wherein said compound is a therapeutic compound that is effective to treat one or more afflictions selected from the group consisting of cancer and infectious diseases.
Claim 24
Dependent Claim
26 words
A pharmaceutical composition according to claim 20 wherein said compound targets a cell selected from the group consisting of a cancerous cell and a pathogen-infected cell.
Claim 25
Dependent Claim
25 words
A pharmaceutical composition according to claim 20 that is effective to transfect cells of interest using molar ratios of agent:compound of between 5:1 and 30:1.
Claim 26
Dependent Claim
28 words
A pharmaceutical composition according to claim 20 in aqueous form in which the compound is present at a concentration of between about 0.1 uM and about 100 uM.
Claim 27
Dependent Claim
29 words
A pharmaceutical composition according to claim 20 in aqueous form in which the compound is present at a molar concentration of between about 1 uM and about 20 uM.
Claim 28
Dependent Claim
159 words
A transfection agent comprising a peptide of between about 16 to 30 amino acid residues, said peptide comprising: a) a hydrophobic domain comprising at least four amino acid residues, wherein the hydrophobic domain comprises three to five aromatic amino acid residues and a first hydrophobic locus spaced by at least one amino acid residue from a second hydrohpobic locus, wherein each of the first and second hydrophobic loci comprise one or more hydrophobic amino acid residues independently selected from the group consisting of Phe, Tyr, Trp, Thr, Met, Leu, Val, Ile, Ala, and His; b) a hydrophilic domain comprising up to about twelve amino acid residues, wherein the hydrophilic domain comprises a plurality of basic amino acid residues; c) optionally a spacer sequence between said hydrophobic and said hydrophilic domain, wherein said spacer comprises from between one to about ten amino acid residues; and d) further optionally a functional group conjugated to one or more termini of said peptide.
Claim 29
Dependent Claim
159 words
A transfection agent comprising a peptide of between about 16 to 30 amino acid residues, said peptide comprising: a) a hydrophobic domain comprising at least four amino acid residues, wherein the hydrophobic domain comprises three to five aromatic amino acid residues and a first hydrophobic locus spaced by at least one amino acid residue from a second hydrohpobic locus, wherein each of the first and second hydrophobic loci comprise one or more hydrophobic amino acid residues independently selected from the group consisting of Phe, Tyr, Trp, Thr, Met, Leu, Val, Ile, Ala, and His; b) a hydrophilic domain comprising up to about twelve amino acid residues, wherein the hydrophilic domain comprises a plurality of basic amino acid residues; c) optionally a spacer sequence between said hydrophobic and said hydrophilic domain, wherein said spacer comprises from between one to about ten amino acid residues; and d) further optionally a functional group conjugated to one or more termini of said peptide.
Claim 30
Dependent Claim
28 words
A transfection agent according to claim 27 wherein said hydrophilic domain is a cation-rich sequence comprised of at least two lysine residues within a span of seven residues.
Claim 31
Dependent Claim
32 words
The transfection agent of claim 27 wherein at least two of said plurality of aromatic amino acid residues occur in a pair in the first hydrophobic locus or the second hydrophobic locus.
Claim 32
Dependent Claim
22 words
The transfection agent of claim 28 wherein two or more of said at least two lysine residues are adjacent to one another.
Claim 33
Dependent Claim
24 words
The transfection agent of claim 27 wherein said plurality of aromatic amino acid residues is between three and five aromatic amino acid residues inclusive.
Claim 34
Dependent Claim
20 words
The transfection agent of claim 27 wherein said plurality of aromatic amino acid residues comprises at least two tryptophan residues.
Claim 35
Dependent Claim
49 words
The transfection agent of claim 27 comprising two pairs of aromatic amino acid residues, the first pair being disposed in the first hydrophobic locus and the second pair being disposed in the second hydrophobic locus, and wherein said first and second hydrophobic loci are separated by two amino acids.
Claim 36
Dependent Claim
23 words
The transfection agent of claim 33 wherein said two amino acids separating the first and second hydrophobic loci consist of hydrophilic amino acids.
Claim 37
Dependent Claim
22 words
The transfection agent of claim 34 wherein said hydrophilic amino acids separating the first and second hydrophobic loci are Glu and Thr.
Claim 38
Dependent Claim
14 words
A transfection agent according to claim 27 wherein said peptide is a synthetic peptide.
Claim 39
Dependent Claim
34 words
A transfection agent according to claim 27 wherein said optional spacer sequence comprises one or more amino acid residues selected from the group consisting of proline, glycine, tyrosine, serine, glutamine, and non-charged amino acids.
Claim 40
Dependent Claim
37 words
A transfection agent according to claim 27 wherein said hydrophobic domain comprises a motif (Trp/Tyr)-(Trp/Tyr)-Xaa-Xaa-(Trp/Tyr), wherein the first and second hydrophobic loci are spaced by two amino acids, Xaa-Xaa, wherein each Xaa is a hydrophilic amino acid.
Claim 41
Dependent Claim
41 words
A transfection agent according to claim 27 wherein said agent is covalently affixed to a compound selected from the group consisting of a nucleic acid, a peptide, a protein, an antibody, and a derivative or analog of any of the foregoing.
Claim 42
Dependent Claim
41 words
A transfection agent according to claim 27 wherein said agent is non-covalently complexed with a compound selected from the group consisting of a nucleic acid, a peptide, a protein, an antibody, and a derivative or analog of any of the foregoing.
Claim 43
Dependent Claim
55 words
A transfection agent according to claim 27 that further comprises a functional group covalently attached to a terminus of said peptide, wherein the functional group is selected from the group consisting of a cysteamine group, a methyl group, and an alkyl group, and, with respect to the amino terminus of said peptide, an acyl group.
Claim 44
Dependent Claim
43 words
A transfection agent according to claim 27 wherein said hydrophilic domain comprises the sequence Lys-Arg-Lys, and wherein said agent further comprises a spacer sequence that comprises at least three amino acid residues of which at least one is a proline or glutamine residue.
Claim 45
Dependent Claim
38 words
A transfection agent according to claim 27 that further comprises a functional group to which is covalently conjugated a molecule selected from the group consisting of a stabilizer, a coupler, a dye, a ligand, and an enzymatic substrate.
Claim 46
Dependent Claim
46 words
A commercial transfection kit comprising at least one transfection agent according to claim 27 in either aqueous or lyophilized form, said kit further comprising one or more components selected from the group consisting of buffers, positive controls, cells to be transfected, phospholipids, and instructions for use.
Claim 47
Dependent Claim
12 words
A pharmaceutical composition comprising a transfection agent according to claim 27 .
Claim 48
Dependent Claim
23 words
A pharmaceutical composition according to claim 45 wherein said transfection agent is non-covalently complexed with a compound to be delivered to a cell.
Claim 49
Dependent Claim
26 words
A pharmaceutical composition according to claim 46 wherein said compound comprises a member selected from the group consisting of a diagnostic compound and a therapeutic compound.
Claim 50
Dependent Claim
33 words
A pharmaceutical composition according to claim 46 wherein said compound is a therapeutic compound that is effective to treat one or more afflictions selected from the group consisting of cancer and infectious diseases.
Claim 51
Dependent Claim
26 words
A pharmaceutical composition according to claim 46 wherein said compound targets a cell selected from the group consisting of a cancerous cell and a pathogen-infected cell.
Claim 52
Dependent Claim
25 words
A pharmaceutical composition according to claim 46 that is effective to transfect cells of interest using molar ratios of agent:compound of between 5:1 and 30:1.
Claim 53
Dependent Claim
28 words
A phannaceutical composition according to claim 46 in aqueous form in which the compound is present at a concentration of between about 0.1 uM and about 100 uM.
Claim 54
Dependent Claim
29 words
A pharmaceutical composition according to claim 46 in aqueous form in which the compound is present at a molar concentration of between about 1 uM and about 20 uM.
Claim 55
Dependent Claim
159 words
A transfection agent comprising a peptide of between about 16 to 30 amino acid residues, said peptide comprising: a) a hydrophobic domain comprising at least four amino acid residues, wherein the hydrophobic domain comprises a plurality of aromatic amino acid residues and a first hydrophobic locus spaced by at least one amino acid residue from a second hydrohpobic locus, wherein each of the first and second hydrophobic loci comprise one or more hydrophobic amino acid residues independently selected from the group consisting of Phe, Tyr, Trp, Thr, Met, Leu, Val, Ile, Ala, and His; b) a hydrophilic domain comprising up to about 12 amino acid residues, wherein the hydrophilic domain comprises a plurality of basic amino acid residues; c) optionally a spacer sequence between said hydrophobic and said hydrophilic domains, wherein said spacer comprises from between one to about ten amino acid residues; and d) further optionally a functional group conjugated to one or more termini of said peptide.
Claim 56
Dependent Claim
159 words
A transfection agent comprising a peptide of between about 16 to 30 amino acid residues, said peptide comprising: a) a hydrophobic domain comprising at least four amino acid residues, wherein the hydrophobic domain comprises a plurality of aromatic amino acid residues and a first hydrophobic locus spaced by at least one amino acid residue from a second hydrohpobic locus, wherein each of the first and second hydrophobic loci comprise one or more hydrophobic amino acid residues independently selected from the group consisting of Phe, Tyr, Trp, Thr, Met, Leu, Val, Ile, Ala, and His; b) a hydrophilic domain comprising up to about 12 amino acid residues, wherein the hydrophilic domain comprises a plurality of basic amino acid residues; c) optionally a spacer sequence between said hydrophobic and said hydrophilic domains, wherein said spacer comprises from between one to about ten amino acid residues; and d) further optionally a functional group conjugated to one or more termini of said peptide.
Claim 57
Dependent Claim
28 words
A transfection agent according to claim 53 wherein said hydrophilic domain is a cation-rich sequence comprised of at least two lysine residues within a span of seven residues.
Claim 58
Dependent Claim
32 words
The transfection agent of claim 53 wherein at least two of said plurality of aromatic amino acid residues occur in a pair in the first hydrophobic locus or the second hydrophobic locus.
Claim 59
Dependent Claim
22 words
The transfection agent of claim 54 wherein two or more of said at least two lysine residues are adjacent to one another.
Claim 60
Dependent Claim
24 words
The transfection agent of claim 53 wherein said plurality of aromatic amino acid residues is between three and five aromatic amino acid residues inclusive.
Claim 61
Dependent Claim
20 words
The transfection agent of claim 53 wherein said plurality of aromatic amino acid residues comprises at least two tryptophan residues.
Claim 62
Dependent Claim
49 words
The transfection agent of claim 53 comprising two pairs of aromatic amino acid residues, the first pair being disposed in the first hydrophobic locus and the second pair being disposed in the second hydrophobic kcus, and wherein said first and second hydrophobic loci are separated by two amino acids.
Claim 63
Dependent Claim
23 words
The transfection agent of claim 59 wherein said two amino acids separating the first and second hydrophobic loci consist of hydrophilic amino acids.
Claim 64
Dependent Claim
22 words
The transfection agent of claim 60 wherein said hydrophilic amino acids separating the first and second hydrophobic loci are Glu and Thr.
Claim 65
Dependent Claim
14 words
A transfection agent according to claim 53 wherein said peptide is a synthetic peptide.
Claim 66
Dependent Claim
34 words
A transfection agent according to claim 53 wherein said optional spacer sequence comprises one or more amino acid residues selected from the group consisting of proline, glycine, tyrosine, serine, glutamine, and non-charged amino acids.
Claim 67
Dependent Claim
37 words
A transfection agent according to claim 53 wherein said hydrophobic domain comprises a motif (Trp/Tyr)-(Trp/Tyr)-Xaa-Xaa-(Trp/Tyr), wherein the first and second hydrophobic loci are spaced by two amino acids, Xaa-Xaa, wherein each Xaa is a hydrophilic amino acid.
Claim 68
Dependent Claim
40 words
A transfection agent according to claim 53 wherein said agent is covalently affixed to a compound selected from the group consisting of a nucleic acid, a peptide, a protein, an antibody, and a derivative/or analog of any of the foregoing.
Claim 69
Dependent Claim
41 words
A transfection agent according to claim 53 wherein said agent is non-covalently complexed with a compound selected from the group consisting of a nucleic acid, a peptide, a protein, an antibody, and a derivatives or analog of any of the foregoing.
Claim 70
Dependent Claim
55 words
A transfection agent according to claim 53 that further comprises a functional group covalently attached to a terminus of said peptide, wherein the functional group is selected from the group consisting of a cysteanline group, a methyl group, and an alkyl group, and, with respect to the amino terminus of said peptide, an acyl group.
Claim 71
Dependent Claim
43 words
A transfection agent according to claim 53 wherein said hydrophilic domain comprises the sequence Lys-Arg-Lys, and wherein said agent further comprises a spacer sequence that comprises at least three amino acid residues of which at least one is a proline or glutamine residue.
Claim 72
Dependent Claim
38 words
A transfection agent according to claim 53 that further comprises a functional group to which is convalently conjugated a molecule selected from the group consisting of a stabilizer, a coupler, a dye, a ligand, and an enzymatic substrate.
Claim 73
Dependent Claim
46 words
A commercial transfection kit comprising at least one transfection agent according to claim 53 in either aqueous or lyophilized form, said kit further comprising one or more components selected from the group consisting of buffers, positive controls, cells to be transfected, phospholipids, and instructions for use.
Claim 74
Dependent Claim
12 words
A pharmaceutical composition comprising a transfection agent according to claim 53 .
Claim 75
Dependent Claim
23 words
A pharmaceutical composition according to claim 71 wherein said transfection agent is non-covalently complexed with a compound to be delivered to a cell.
Claim 76
Dependent Claim
26 words
A pharmaceutical composition according to claim 72 wherein said compound comprises a member selected from the group consisting of a diagnostic compound and a therapeutic compound.
Claim 77
Dependent Claim
33 words
A pharmaceutical composition according to claim 72 wherein said compound is a therapeutic compound that is effective to treat one or more afflictions selected from the group consisting of cancer and infectious diseases.
Claim 78
Dependent Claim
26 words
A pharmaceutical composition according to claim 72 wherein said compound targets a cell selected from the group consisting of a cancerous cell and a pathogen-infected cell.
Claim 79
Dependent Claim
25 words
A pharmaceutical composition according to claim 72 that is effective to transfect cells of interest using molar ratios of agent:compound of between 5:1 and 30:1.
Claim 80
Dependent Claim
28 words
A pharmaceutical composition according to claim 72 in aqueous form in which the compound is present at a concentration of between about 0.1 uM and about 100 uM.
Claim 81
Dependent Claim
29 words
A pharmaceutical composition according to claim 72 in aqueous form in which the compound is present at a molar concentration of between about 1 uM and about 20 uM.
Claim 82
Dependent Claim
173 words
A transfection agent comprising a peptide of between about 16 to 30 amino acid residues in length, said peptide comprising: a) a hydrophobic domain comprising at least four amino acid residues, wherein the hydrophobic domain comprises a first hydrophobic locus spaced by at least one amino acid residue from a second hydrohpobic locus, wherein each of the first and second hydrophobic loci comprise one or more hydrophobic amino acid residues independently selected from the group consisting of Phe, Tyr, Trp, Thr, Met, Leu, Val, Ile, Ala, and His; b) a hydrophilic domain comprising up to about 12 amino acid residues, wherein the hydrophilic domain comprises a plurality of basic amino acid residues; c) optionally a spacer sequence between said hydrophobic and said hydrophilic domains, wherein said spacer comprises from between one to about ten amino acid residues; and d) further optionally a functional group conjugated to one or more termini of said peptide, wherein the peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO. 1 through 12.
Claim 83
Dependent Claim
173 words
A transfection agent comprising a peptide of between about 16 to 30 amino acid residues in length, said peptide comprising: a) a hydrophobic domain comprising at least four amino acid residues, wherein the hydrophobic domain comprises a first hydrophobic locus spaced by at least one amino acid residue from a second hydrohpobic locus, wherein each of the first and second hydrophobic loci comprise one or more hydrophobic amino acid residues independently selected from the group consisting of Phe, Tyr, Trp, Thr, Met, Leu, Val, Ile, Ala, and His; b) a hydrophilic domain comprising up to about 12 amino acid residues, wherein the hydrophilic domain comprises a plurality of basic amino acid residues; c) optionally a spacer sequence between said hydrophobic and said hydrophilic domains, wherein said spacer comprises from between one to about ten amino acid residues; and d) further optionally a functional group conjugated to one or more termini of said peptide, wherein the peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO. 1 through 12.
Claim 84
Dependent Claim
14 words
A transfection agent according to claim 79 wherein said peptide is a synthetic peptide.
Claim 85
Dependent Claim
41 words
A transfection agent according to claim 79 wherein said agent is covalently affixed to a compound selected from the group consisting of a nucleic acid, a peptide, a protein, an antibody, and a derivative or analog of any of the foregoing.
Claim 86
Dependent Claim
41 words
A transfection agent according to claim 79 wherein said agent is non-covalently complexed with a compound selected from the group consisting of a nucleic acid, a peptide, a protein, an antibody, and a derivatives or analog of any of the foregoing.
Claim 87
Dependent Claim
55 words
A transfection agent according to claim 79 that further comprises a functional group covalently attached to a terminus of said peptide, wherein the functional group is selected from the group consisting of a cysteamine group, a methyl group, and an alkyl group, and, with respect to the amino terminus of said peptide, an acyl group.
Claim 88
Dependent Claim
43 words
A transfection agent according to claim 79 wherein said hydrophilic domain comprises the sequence Lys-Arg-Lys, and wherein said agent further comprises a spacer sequence that comprises at least three amino acid residues of which at least one is a proline or glutamine residue.
Claim 89
Dependent Claim
38 words
A transfection agent according to claim 79 that further comprises a functional group to which is covelently conjugated a molecule selected from the group consisting of a stabilizer, a coupler, a dye, a ligand, and an enzymatic substrate.
Claim 90
Dependent Claim
46 words
A commercial transfection kit comprising at least one transfection agent according to claim 79 in either aqueous or lyophilized form, said kit further comprising one or more components selected from the group consisting of buffers, positive controls, cells to be transfected, phospholipids, and instructions for use.
Claim 91
Dependent Claim
12 words
A pharmaceutical composition comprising a transfection agent according to claim 79 .
Claim 92
Dependent Claim
23 words
A pharmaceutical composition according to claim 87 wherein said transfection agent is non-covalently complexed with a compound to be delivered to a cell.
Claim 93
Dependent Claim
26 words
A pharmaceutical composition according to claim 88 wherein said compound comprises a member selected from the group consisting of a diagnostic compound and a therapeutic compound.
Claim 94
Dependent Claim
33 words
A pharmaceutical composition according to claim 88 wherein said compound is a therapeutic compound that is effective to treat one or more afflictions selected from the group consisting of cancer and infectious diseases.
Claim 95
Dependent Claim
26 words
A pharmaceutical composition according to claim 88 wherein said compound targets a cell selected from the group consisting of a cancerous cell and a pathogen-infected cell.
Claim 96
Dependent Claim
25 words
A pharmaceutical composition according to claim 88 that is effective to transfect cells of interest using molar ratios of agent:compound of between 5:1 and 30:1.
Claim 97
Dependent Claim
28 words
A pharmaceutical composition according to claim 88 in aqueous form in which the compound is present at a concentration of between about 0.1 uM and about 100 uM.
Claim 98
Dependent Claim
29 words
A pharmaceutical composition according to claim 88 in aqueous form in which the compound is present at a molar concentration of between about 1 uM and about 20 uM.
Claim 99
Dependent Claim
197 words
A commercial transfection kit comprising at least one transfection agent comprising a peptide of between about 16 to 30 amino acid residues, said peptide comprising: a) a hydrophobic domain comprising at least four amino acid residues, wherein the hydrophobic domain comprises a first hydrophobic locus spaced by at least one amino acid residue from a second hydrohpobic locus, wherein each of the first and second hydrophobic loci comprise one or more hydrophobic amino acid residues independently selected from the group consisting of Phe, Tyr, Trp, Thr, Met, Leu, Val, Ile, Ala, and His; b) a hydrophilic domain comprising up to about 12 amino acid residues, wherein the hydrophilic domain comprises a plurality of basic amino acid residues; c) optionally a spacer sequence between said hydrophobic and said hydrophilic domains, wherein said spacer comprises from between one to about ten amino acid residues; and d) further optionally a functional group conjugated to one or more termini of said peptide, wherein the transfection agent is in either aqueous or lyophilized form, and wherein said kit further comprises one or more components selected from the group consisting of buffers, positive controls, cells to be transfected, phospholipids, and instructions for use.
Claim 100
Dependent Claim
197 words
A commercial transfection kit comprising at least one transfection agent comprising a peptide of between about 16 to 30 amino acid residues, said peptide comprising: a) a hydrophobic domain comprising at least four amino acid residues, wherein the hydrophobic domain comprises a first hydrophobic locus spaced by at least one amino acid residue from a second hydrohpobic locus, wherein each of the first and second hydrophobic loci comprise one or more hydrophobic amino acid residues independently selected from the group consisting of Phe, Tyr, Trp, Thr, Met, Leu, Val, Ile, Ala, and His; b) a hydrophilic domain comprising up to about 12 amino acid residues, wherein the hydrophilic domain comprises a plurality of basic amino acid residues; c) optionally a spacer sequence between said hydrophobic and said hydrophilic domains, wherein said spacer comprises from between one to about ten amino acid residues; and d) further optionally a functional group conjugated to one or more termini of said peptide, wherein the transfection agent is in either aqueous or lyophilized form, and wherein said kit further comprises one or more components selected from the group consisting of buffers, positive controls, cells to be transfected, phospholipids, and instructions for use.
Claim 101
Dependent Claim
27 words
A kit according to claim 95 wherein said hydrophilic domain is a cation-rich sequence comprised of at least two lysine residues within a span of seven residues.
Claim 102
Dependent Claim
32 words
A kit according to claim 95 wherein at least two of said plurality of aromatic amino acid residues occur in a pair in the first hydrophobic locus or the second hydrophobic locus.
Claim 103
Dependent Claim
22 words
A kit according to claim 96 wherein two or more of said at least two lysine residues are adjacent to one another.
Claim 104
Dependent Claim
24 words
A kit according to claim 95 wherein said plurality of aromatic amino acid residues is between three and five aromatic amino acid residues inclusive.
Claim 105
Dependent Claim
20 words
A kit according to claim 95 wherein said plurality of aromatic amino acid residues comprises at least two tryptophan residues.
Claim 106
Dependent Claim
49 words
A kit according to claim 95 comprising two pairs of aromatic amino acid residues, the first pair being disposed in the first hydrophobic locus and the second pair being disposed in the second hydrophobic locus, and wherein said first and second hydrophobic loci are separated by two amino acids.
Claim 107
Dependent Claim
23 words
A kit according to claim 101 wherein said two amino acids separating the first and second hydrophobic loci consist of hydrophilic amino acids.
Claim 108
Dependent Claim
22 words
A kit according to claim 102 wherein said hydrophilic amino acids separating the first and second hydrophobic loci are Glu and Thr.
Claim 109
Dependent Claim
13 words
A kit according to claim 95 wherein said peptide is a synthetic peptide.
Claim 110
Dependent Claim
33 words
A kit according to claim 95 wherein said optional spacer sequence comprises one or more amino acid residues selected from the group consisting of proline, glycine, tyrosine, serine, glutamine, and non-charged amino acids.
Claim 111
Dependent Claim
36 words
A kit according to claim 95 wherein said hydrophobic domain comprises a motif (Trp/Tyr)-(Trp/Tyr)-Xaa-Xaa-(Trp/Tyr), wherein the first and second hydrophobic loci are spaced by two amino acids, Xaa-Xaa, wherein each Xaa is a hydrophilic amino acid.
Claim 112
Dependent Claim
40 words
A kit according to claim 95 wherein said agent is covalently affixed to a compound selected from the group consisting of a nucleic acid, a peptide, a protein, an antibody, and a derivatives or analog of any of the foregoing.
Claim 113
Dependent Claim
40 words
A kit according to claim 95 wherein said agent is non-covalently complexed with a compound selected from the group consisting of a nucleic acid, a peptide, a protein, an antibody, and a derivatives or analog of any of the foregoing.
Claim 114
Dependent Claim
54 words
A kit according to claim 95 that further comprises a functional group covalently attached to a terminus of said peptide, wherein the functional group is selected from the group consisting of a cysteamine group, a methyl group, and an alkyl group, and, with respect to the amino terminus of said peptide, an acyl group.
Claim 115
Dependent Claim
42 words
A kit according to claim 95 wherein said hydrophilic domain comprises the sequence Lys-Arg-Lys, and wherein said agent further comprises a spacer sequence that comprises at least three amino acid residues of which at least one is a proline or glutamine residue.
Claim 116
Dependent Claim
37 words
A kit according to claim 95 that further comprises a functional group to which is covelently conjugated a molecule selected from the group consisting of a stabilizer, a coupler, a dye, a ligand, and an enzymatic substrate.
Claim 117
Dependent Claim
16 words
A kit according to claim 95 wherein the transfection agent is formulated as a pharmaceutical composition.
Claim 118
Dependent Claim
22 words
A kit according to claim 112 wherein said transfection agent is non-covalently complexed with a compound to be delivered to a cell.
Claim 119
Dependent Claim
25 words
A kit according to claim 113 wherein said compound comprises a member selected from the group consisting of a diagnostic compound and a therapeutic compound.
Claim 120
Dependent Claim
32 words
A kit according to claim 113 wherein said compound is a therapeutic compound that is effective to treat one or more afflictions selected from the group consisting of cancer and infectious diseases.
Claim 121
Dependent Claim
25 words
A kit according to claim 113 wherein said compound targets a cell selected from the group consisting of a cancerous cell and a pathogen-infected cell.
Claim 122
Dependent Claim
26 words
A kit according to claim 113 wherein said compound is effective to transfect cells of interest using molar ratios of agent:compound of between 5:1 and 30:1.
Claim 123
Dependent Claim
30 words
A kit according to claim 113 wherein said composition is in aqueous form and the compound is present at a concentration of between about 0.1 uM and about 100 uM.
Claim 124
Dependent Claim
31 words
A kit according to claim 113 wherein said composition is in aqueous form and the compound is present at a molar concentration of between about 1 uM and about 20 uM.
References Cited (Backward Citations)
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