Patent US6117632A successfully refreshed from Google Patents.
US6117632A
Expired / Lapsed
US
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Peptides which enhance transport across tissues and methods of identifying and using the same
Assignee: Novo Nordisk AS
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Filing: 1996-11-08
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Granted: 2000-09-12
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Expires: 2016-11-08
Abstract
Abstract A method of identifying a peptide which permits or facilitates the transport of an active agent through a human or animal tissue. A predetermined amount of phage from a random phage library or preselected phage library is plated unto or brought into contact with a first side, preferably the apical side, of a tissue sample or polarized tissue cell culture. At a predetermined time, the phage which is transported to a second side of the tissue opposite the first side, preferably the basolateral side, is harvested to select transported phage. This modified phage is amplified in a host. This cycle of events is repeated (using the transported phage produced in the most recent cycle) a predetermined number of times to obtain a selected phage library containing phage which can be transported from the first side to the second side. Lastly, the identity of at least one peptide coded by phage in the selected phage library is determined to identify a peptide which permits or facilitates the transport of an active agent through a human or animal tissue.
Inventors
- 👤 Daniel Joseph O'Mahony
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 17:34
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/68; B; B82; B82Y; B82Y5/00; C; C07; C07K; C07K14/00; C07K14/435; C07K14/705
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
189 words
A method of identifying a peptide which permits or facilitates the transport of an active agent through a human or animal tissue, comprising the steps of: (a) contacting a predetermined amount of phage from a random phage library or a preselected phage library to a first side of a tissue sample or polarized tissue cell culture; (b) at a predetermined time harvesting phage which is transported to a second side of the tissue sample or culture to select transported phage, which second side is opposite the first side; (c) amplifying the transported phage in a host; (d) repeating in order step (a) using the transported phage obtained in step (b) and amplified in step (c) and steps (b) and (c) a predetermined number of times to obtain a selected phage library containing phage which can be transported from the first side of the tissue sample or culture to the second side; and (e) determining the identity of at least one peptide coded by phage in the selected phage library to identify a peptide which permits or facilitates the transport of an active agent through a human or animal tissue.
Claim 2
Independent Claim
189 words
A method of identifying a peptide which permits or facilitates the transport of an active agent through a human or animal tissue, comprising the steps of: (a) contacting a predetermined amount of phage from a random phage library or a preselected phage library to a first side of a tissue sample or polarized tissue cell culture; (b) at a predetermined time harvesting phage which is transported to a second side of the tissue sample or culture to select transported phage, which second side is opposite the first side; (c) amplifying the transported phage in a host; (d) repeating in order step (a) using the transported phage obtained in step (b) and amplified in step (c) and steps (b) and (c) a predetermined number of times to obtain a selected phage library containing phage which can be transported from the first side of the tissue sample or culture to the second side; and (e) determining the identity of at least one peptide coded by phage in the selected phage library to identify a peptide which permits or facilitates the transport of an active agent through a human or animal tissue.
Claim 3
Dependent Claim
15 words
The method of claim 1, wherein step (d) is repeated from 0 to 30 times.
Claim 4
Dependent Claim
59 words
The method of claim 1, wherein the tissue sample is selected from the group consisting of duodenum, jejunum, ileum, ascending colon, transverse colon, descending colon, pelvic colon, vascular endothelium which lines the vascular system, vascular endothelium of the blood brain barrier, vascular smooth muscle, alveolar, liver, kidney, bone marrow, heart, spleen, pancreas, thymus, brain, neuronal or retinal eye tissue.
Claim 5
Dependent Claim
20 words
The method of claim 1, wherein the tissue sample comprises epithelial cells lining the lumenal side of the gastrointestinal tract.
Claim 6
Dependent Claim
14 words
The method of claim 4, wherein the tissue sample is derived from the colon.
Claim 7
Dependent Claim
35 words
The method of claim 1, wherein the polarized tissue cell culture is cultured from gastro-intestinal tract epithelial cells, alveolar cells, endothelial cells of the blood-brain barrier, vascular smooth muscle cells, Caco-2 cells, or T-84 cells.
Claim 8
Dependent Claim
27 words
The method of claim 1, wherein the first side of the tissue sample or culture is the apical side and the second side is the basolateral side.
Claim 9
Dependent Claim
14 words
The method of claim 1, wherein the active agent is a drug or antigen.
Claim 10
Dependent Claim
14 words
The method of claim 1, wherein the active agent is a nano- or microparticle.
Claim 11
Dependent Claim
24 words
The method of claim 9, wherein the peptide is coated onto or adsorbed onto or covalently bonded to the surface of the nano-or microparticle.
Claim 12
Dependent Claim
15 words
The method of claim 9, wherein the nano- or microparticle is formed from the peptide.
Claim 13
Dependent Claim
17 words
The method claim 9, wherein the nano- or microparticle is a drug-loaded or drug-encapsulated nano- or microparticle.
Claim 14
Independent Claim
181 words
A method of identifying a peptide which permits or facilitates the transport of an active agent through a human or animal tissue, comprising the steps of: (a) contacting a predetermined amount of phage from a random phage library or a preselected phage library to the apical side of a tissue sample or polarized tissue cell culture; (b) at a predetermined time harvesting phage which is transported to the basolateral side of the tissue sample or culture to select transported phage; (c) amplifying the transported phage in a host; (d) repeating in order step (a) using the transported phage obtained in step (b) and amplified in step (c) and steps (b) and (c) a predetermined number of times to obtain a selected phage library containing phage which can be transported from the apical side of the tissue sample or culture to the basolateral side; and (e) determining the identity of at least one peptide coded by phage in the selected phage library to identify a peptide which permits or facilitates the transport of an active agent through a human or animal tissue.
Claim 15
Independent Claim
181 words
A method of identifying a peptide which permits or facilitates the transport of an active agent through a human or animal tissue, comprising the steps of: (a) contacting a predetermined amount of phage from a random phage library or a preselected phage library to the apical side of a tissue sample or polarized tissue cell culture; (b) at a predetermined time harvesting phage which is transported to the basolateral side of the tissue sample or culture to select transported phage; (c) amplifying the transported phage in a host; (d) repeating in order step (a) using the transported phage obtained in step (b) and amplified in step (c) and steps (b) and (c) a predetermined number of times to obtain a selected phage library containing phage which can be transported from the apical side of the tissue sample or culture to the basolateral side; and (e) determining the identity of at least one peptide coded by phage in the selected phage library to identify a peptide which permits or facilitates the transport of an active agent through a human or animal tissue.
Claim 16
Independent Claim
183 words
A method of identifying a peptide motif which when present in a peptide permits or facilitates the transport of an active agent through a human or animal tissue, comprising the steps of: (a) contacting a predetermined amount of phage from a random phage library to the apical side of a tissue sample or polarized tissue cell culture sample; (b) at a predetermined time harvesting phage which is transported to the basolateral side of the tissue to select modified phage; (c) amplifying the modified phage in a host; (d) repeating in order step (a) using the modified phage obtained in step (b) and amplified in step (c) and steps (b) and (c) a predetermined number of times to obtain a selected phage library containing phage which can be transported from the first side to the second side; and (e) determining the identity of a plurality of peptides coded by phage in the selected phage library to identify a peptide motif common to at least two of the peptides which permit or facilitate the transport of an active agent through a human or animal tissue.
Claim 17
Independent Claim
183 words
A method of identifying a peptide motif which when present in a peptide permits or facilitates the transport of an active agent through a human or animal tissue, comprising the steps of: (a) contacting a predetermined amount of phage from a random phage library to the apical side of a tissue sample or polarized tissue cell culture sample; (b) at a predetermined time harvesting phage which is transported to the basolateral side of the tissue to select modified phage; (c) amplifying the modified phage in a host; (d) repeating in order step (a) using the modified phage obtained in step (b) and amplified in step (c) and steps (b) and (c) a predetermined number of times to obtain a selected phage library containing phage which can be transported from the first side to the second side; and (e) determining the identity of a plurality of peptides coded by phage in the selected phage library to identify a peptide motif common to at least two of the peptides which permit or facilitate the transport of an active agent through a human or animal tissue.
References Cited (Backward Citations)
US5672584A
Cited
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| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 2003-09-12 | 2004-03-13 | 2004-09-12 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2007-09-12 | 2008-03-13 | 2008-09-12 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2011-09-12 | 2012-03-13 | 2012-09-12 | $7700.00 | Window Closed / Lapsed |