Patent US4455290A successfully refreshed from Google Patents.
US4455290A
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Inhibition of fibrin polymerization by a peptide isolated from fibrin Fragment D1
Assignee: Research Corp
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Filing: 1981-04-02
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Granted: 1984-06-19
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Expires: 2001-04-02
Abstract
Abstract A purified peptide isolated from fibrinogen Fragment D 1 and having the amino acid sequence Thr-Arg-Trp-Tyr-Ser-Met-Lys-Lys-Thr-Thr-Met-Lys-Ile-Ile-Pro-Phe-Asn-Arg-Leu-Thr-Ile-Gly-Glu-Gly-Gln-Gln-His-His-Leu-Gly-Gly-Ala-Lys-Gln-Ala-Gly-Asp-Val. The peptide is isloated by degrading Fragment D 1 of fibrinogen with plasmin followed by separation of the resulting peptides on the basis of molecular weight and affinity for bound fibrin monomer. The purified peptide is useful as an anticoagulant and, when suitably labeled with a gamma-emitting radioisotope, as a thrombus imaging agent.
Inventors
- 👤 Stephanie A. Olexa
- 👤 Andrei Z. Budzynski
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 21:25
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/745; C07K14/75; A; A61; A61K; A61K38/00
Claims Analysis
Total Claims: 28
Claim 1
Independent Claim
31 words
A purified peptide having the amino acid sequence (a) Gln-Ala-Gly-Asp-Val, (b) Thr-Arg-Trp-Tyr-Ser-Met-Lys-Lys-Thr-Thr-Met-Lys-Ile-Ile-Pro-Phe-Asn-Arg-Leu-Thr-Ile-Gly-Glu-Gly-Gln-Gln-His-His-Leu-Gly-Gly-Ala-Lys-Gln-Ala-Gly-Asp-Val or (c) a purified peptide having an amino acid sequence intermediate between said amino acid sequences (a) and (b).
Claim 2
Dependent Claim
31 words
A purified peptide having the amino acid sequence (a) Gln-Ala-Gly-Asp-Val, (b) Thr-Arg-Trp-Tyr-Ser-Met-Lys-Lys-Thr-Thr-Met-Lys-Ile-Ile-Pro-Phe-Asn-Arg-Leu-Thr-Ile-Gly-Glu-Gly-Gln-Gln-His-His-Leu-Gly-Gly-Ala-Lys-Gln-Ala-Gly-Asp-Val or (c) a purified peptide having an amino acid sequence intermediate between said amino acid sequences (a) and (b).
Claim 3
Dependent Claim
21 words
A purified peptide having the amino acid sequence X-Gln-Ala-Gly-Asp-Val, wherein X represents Ala-Lys-, Lys-, or the absence of an amino acid.
Claim 4
Dependent Claim
21 words
A purified peptide having the amino acid sequence X-Gln-Ala-Gly-Asp-Val, wherein X represents Ala-Lys-, Lys-, or the absence of an amino acid.
Claim 5
Dependent Claim
42 words
A purified peptide having substantially the amino acid sequence disclosed in claim 1(a), 1(c), or 2 wherein said purified peptide has a binding affinity for fibrin of at least 5% of that demonstrated by the amino acid sequence disclosed in claim 1(b).
Claim 6
Dependent Claim
9 words
A purified peptide having the amino acid sequence Thr-Arg-Trp-Tyr-Ser-Met-Lys-Lys-Thr-Thr-Met-Lys-Ile-Ile-Pro-Phe-Asn-Arg-Leu-Thr-Ile-Gly-Glu-Gly-Gln-Gln-His-His-Leu-Gly-Gly-Ala-Lys-Gln-Ala-Gly-Asp-Val.
Claim 7
Dependent Claim
9 words
A purified peptide having the amino acid sequence Thr-Arg-Trp-Tyr-Ser-Met-Lys-Lys-Thr-Thr-Met-Lys-Ile-Ile-Pro-Phe-Asn-Arg-Leu-Thr-Ile-Gly-Glu-Gly-Gln-Gln-His-His-Leu-Gly-Gly-Ala-Lys-Gln-Ala-Gly-Asp-Val.
Claim 8
Independent Claim
71 words
A method of preparing a peptide inhibitor of fibrin monomer polymerization, comprising the steps of: degrading Fragment D 1 of fibrinogen with plasmin to form plasmin degradation products; separating said peptide inhibitor from plasmin degradation products of different molecular weights to give a partially purified peptide inhibitor; and isolating said peptide inhibitor from other peptides in said partially purified inhibitor by means of said peptide inhibitor's binding affinity for fibrin monomer.
Claim 9
Independent Claim
71 words
A method of preparing a peptide inhibitor of fibrin monomer polymerization, comprising the steps of: degrading Fragment D 1 of fibrinogen with plasmin to form plasmin degradation products; separating said peptide inhibitor from plasmin degradation products of different molecular weights to give a partially purified peptide inhibitor; and isolating said peptide inhibitor from other peptides in said partially purified inhibitor by means of said peptide inhibitor's binding affinity for fibrin monomer.
Claim 10
Dependent Claim
11 words
The method of claim 5, wherein said separating comprises gel filtration.
Claim 11
Dependent Claim
16 words
The method of claim 6, wherein said gel filtration is carried out using dextran gel beads.
Claim 12
Dependent Claim
24 words
The method of claim 7, wherein said dextran gel beads have a dry diameter of 40-120μ and are eluted with an acetic acid solution.
Claim 13
Dependent Claim
11 words
The method of claim 5, wherein said separating comprises affinity chromatography.
Claim 14
Dependent Claim
20 words
The method of claim 9, wherein said affinity chromatography is carried out with fibrin monomer insolubilized on cyanogen bromide-activated agarose.
Claim 15
Dependent Claim
46 words
The method of claim 10, wherein said isolating comprises: applying said partially purified peptide inhibitor to said agarose in a buffer having a pH between 6 and 8; washing said agarose with said buffer; and eluting said peptide inhibitor with an acid, base or high-salt solution.
Claim 16
Dependent Claim
17 words
A peptide inhibitor of fibrin monomer polymerization prepared by the process of claims 5, 8 or 11.
Claim 17
Independent Claim
42 words
A method of inhibiting blood clotting comprising the steps of administering to a human or animal an amount of the peptide inhibitor of fibrin monomer polymerization prepared by the process of claims 5, 8 or 11, sufficient to increase blood clotting time.
Claim 18
Independent Claim
33 words
A method of inhibiting blood clotting comprising adding to a container of blood an amount of the peptide inhibitor of fibrin monomer polymerization prepared by the process of claims 5, 8 or 11.
Claim 19
Dependent Claim
31 words
A method of inhibiting blood clotting comprising the step of administering to a human or animal an amount of the purified peptide of claim 1 sufficient to increase blood clotting time.
Claim 20
Independent Claim
52 words
A method of detecting a thrombus comprising the step of administering to a human or animal an amount of the peptide inhibitor of fibrin monomer polymerization prepared by the process of claims 5, 8 or 11 labeled with a gamma-emitting radioisotope sufficient to allow external detection of the location of said thrombus.
Claim 21
Dependent Claim
41 words
A method of detecting a thrombus comprising the step of administering to a human or animal an amount of the purified peptide of claim 1 labeled with a gamma-emitting radioisotope sufficient to allow external detection of the location of said thrombus.
Claim 22
Dependent Claim
28 words
A purified peptide having the amino acid sequence described in claim 1, wherein said purified peptide is labeled with a gamma-emitting radioactive isotope or an X-ray contrast agent.
Claim 23
Dependent Claim
33 words
The peptide inhibitor of fibrin monomer polymerization prepared by the process of claims 5, 8 or 12 wherein said peptide inhibitor is labeled with a gamma-emitting radioactive isotope or an X-ray contrast agent.
Claim 24
Dependent Claim
20 words
A composition suitable for intravenous injection, comprising: the purified peptide of claims 1 or 18; and a pharmaceutically acceptable carrier.
Claim 25
Dependent Claim
29 words
A composition suitable for intravenous injection comprising: a peptide inhibitor of fibrin monomer polymerization prepared by the process of claims 5, 8 or 12; and a pharmaceutically acceptable carrier.
Claim 26
Dependent Claim
42 words
A radioactive composition suitable for intravenous injection, comprising: a peptide inhibitor of fibrin monomer polymerization prepared by the process of claim 5, 8 or 11 wherein a gamma-emitting radioactive isotope is covalently attached to said purified peptide; and a pharmaceutically acceptable carrier.
Claim 27
Independent Claim
44 words
A method of detecting a thrombus comprising the step of administering to a human or animal an amount of the peptide inhibitor of fibrin monomer polymerization of claims 5, 8 or 11 labeled with an X-ray contrast agent sufficient to cast an X-ray shadow.
Claim 28
Dependent Claim
40 words
A method of detecting a thrombus comprising the step of administering to a human or animal an amount of the purified peptide of claim 1 labeled with an X-ray contrast agent sufficient to cast an X-ray shadow of said thrombus.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1987-06-19 | 1987-12-19 | 1988-06-19 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-06-19 | 1991-12-19 | 1992-06-19 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-06-19 | 1995-12-19 | 1996-06-19 | $7700.00 | Window Closed / Lapsed |