Claims
- 1. A method for inhibiting the binding of a human ICAM-1 molecule to a human LFA-1 molecule comprising the steps of:
- (a) providing a blocking peptide and a modulator peptide, wherein the blocking peptide has an binding affinity for the ICAM-1 molecule, and, when bound to the ICAM-1 molecule, inhibits the binding of the ICAM-1 molecule to the LFA-1 molecule; and
- a modulator peptide, wherein the modulator peptide, when bound to the ICAM-1 molecule, increases the binding affinity of the blocking peptide for the ICAM-1 molecule by inducing a conformational change in the ICAM-1 molecule,
- wherein the blocking peptide and the modulator peptide each comprise an amino acid sequence included in the amino acid sequence of the extracellular segment of the LFA-1 molecule and each have molecular weights of under 20 kilodaltons thereby making them nonimmunogenic in a human; and
- (b) contacting the blocking peptide and the modulator peptide with the ICAM-1 molecule, thereby inhibiting the binding of the ICAM-1 molecule to the LFA-1 molecule.
- 2. The method of claim 1, wherein the blocking peptide comprises an amino acid sequence selected from the group consisting of Sequence ID Nos. 16, 17, and 18.
- 3. The method of claim 1, wherein the modulator peptide comprises Sequence ID No. 16.
- 4. The method of claim 1, wherein the blocking peptide is prepared by a method comprising the steps of:
- (a) identifying a first protein domain of the LFA-1 molecule wherein the first protein domain is capable of binding to the ICAM-1 molecule; and
- (b) synthesizing a blocking peptide comprising an amino acid sequence included in the first protein domain,
- wherein, in the antibody-binding assay of Example 2, the fluorescence intensity value calculated using a first sample lacking the blocking peptide is at least 10% higher than the fluorescence intensity value calculated using a second sample including the blocking peptide,
- wherein the modulator peptide is prepared by a method comprising the steps of:
- (c) identifying a second protein domain of the LFA-1 molecule wherein the second protein domain is capable of binding to the ICAM-1 molecule; and
- (d) synthesizing the modulator peptide comprising an amino acid sequence included in the second protein domain,
- wherein, in the binding assay of Example 2, the fluorescence intensity value calculated using a first sample lacking the modulator peptide is at least 10% lower than the fluorescence intensity value calculated using a second sample including the modulator peptide.
- 5. The method of claim 4, wherein the blocking peptide comprises an amino acid sequence selected from the group consisting of Sequence ID Nos. 16, 17, and 18.
- 6. The method of claim 4, wherein the modulator peptide comprises Sequence ID No.16.
- 7. A method for inhibiting the binding of a human LFA-1 molecule to a human ICAM-1 molecule comprising the steps of:
- (a) providing a blocking peptide and a modulator peptide, wherein the blocking peptide has an initial binding affinity for the LFA-1 molecule, and, when bound to the LFA-1 molecule, inhibits the binding of the LFA-1 molecule to the ICAM-1 molecule; and
- a modulator peptide, wherein the modulator peptide, when bound to the LFA-1 molecule, increases the binding affinity of the blocking peptide for the LFA-1 molecule by inducing a conformational change in the LFA-1 molecule,
- wherein the blocking peptide and the modulator peptide each comprise an amino acid sequence included in the amino acid sequence of the extracellular segment of the ICAM-1 molecule and each have molecular weights of under 20 kilodaltons thereby making them nonimmunogenic in a human; and
- (b) contacting the blocking peptide and the modulator peptide with the LFA-1 molecule, thereby inhibiting the binding of the LFA-1 molecule to the ICAM-1 molecule.
- 8. The method of claim 7, wherein the blocking peptide comprises an amino acid sequence selected from the group consisting of Sequence ID Nos. 6, 10, 11, and 12.
- 9. The method of claim 7, wherein the modulator peptide comprises Sequence ID No. 11.
- 10. The method of claim 7, wherein the blocking peptide is prepared by a method comprising the steps of:
- (a) identifying a first protein domain of the ICAM-1 molecule wherein the first protein domain is capable of binding to the LFA-1 molecule; and
- (b) synthesizing a blocking peptide comprising an amino acid sequence included in the first protein domain,
- wherein, in the antibody-binding assay of Example 2, the fluorescence intensity value calculated using a first sample lacking the blocking peptide is at least 10% higher than the fluorescence intensity value calculated using a second sample including the blocking peptide,
- wherein the modulator peptide is prepared by a method comprising the steps of:
- (c) identifying a second protein domain of the ICAM-1 molecule wherein the second protein domain is capable of binding to the LFA-1 molecule; and
- (d) synthesizing the modulator peptide comprising an amino acid sequence included in the second protein domain,
- wherein, in the binding assay of Example 2, the fluorescence intensity value calculated using a first sample lacking the modulator peptide is at least 10% lower than the fluorescence intensity value calculated using a second sample including the modulator peptide.
- 11. The method of claim 10, wherein the blocking peptide comprises an amino acid sequence selected from the group consisting of Sequence ID Nos. 6,10, 11, and 12.
- 12. The method of claim 10, wherein the modulator peptide comprises Sequence ID No.11.
- 13. Method for inhibiting the interaction between ICAM-1 and LFA-1, said method comprising the steps of:
- (a) providing a blocking peptide comprising a first amino acid sequence of the extracellular segment of one of the human LFA-1 or human ICAM-1 molecule and having a molecular weight of under 20 kilodaltons, said blocking peptide being capable of binding to a target molecule, which is the other of said human LFA-1 or human ICAM-1 molecule, for inhibiting said LFA-1/ICAM-1 interaction,
- said blocking peptide characterized by the property of having a fluorescence intensity value calculated using a first sample lacking said blocking peptide at least 10% higher than the fluorescence intensity value calculated using a second sample including said blocking peptide, in the antibody-binding assay conducted as described in Example 2;
- (b) providing a modulator peptide comprising a second amino acid sequence of the extracellular segment of said one of the human LFA-1 or human ICAM-1 molecule which is different than said first sequence, and having molecular weight of under 20 kilodaltons, said modulator peptide being capable of binding to said target molecule for increasing the affinity of said blocking peptide for said target molecule,
- said modulator peptide characterized by the property of having a fluorescence intensity value calculated using a first sample lacking the modulator peptide of at least 10% lower than the fluorescence intensity value calculated using a second sample including the modulator peptide in the assay of Example 2; and,
- (c) contacting said blocking peptide and said modulator peptide with said target molecule, thereby inhibiting said ICAM-1/LFA-1 interaction.
- 14. The method of claim 13, said one of said human LFA-1 or human ICAM-1 molecule being LFA-1, and said target molecule being ICAM-1.
- 15. The method of claim 13, said one of said human LFA-1 or human ICAM-1 molecule being ICAM-1, and said target molecule being LFA-1.
- 16. The method of claim 13, said blocking peptide being selected from the group consisting of Sequence ID Nos. 6. 17, and 18.
- 17. The method of claim 13, said modulator peptide being selected from the group consisting of Sequence ID Nos. 11 and 16.
- 18. A method of enhancing the interaction between ICAM-1 and LFA-1 comprising:
- (a) providing a modulator peptide comprising an amino acid sequence of the extracellular segment of one of the human LFA-1 or human ICAM-1 molecule, and having a molecular weight of under 20 kilodaltons,
- said modulator peptide characterized by the property of having a fluorescence intensity value calculated using a first sample lacking said modulator peptide of at least 10% lower than the fluorescence intensity value calculated using a second sample including the modulator peptide in the assay of Example 2; and,
- (b) contacting said modulator peptide with the other of said human LFA-1 or human ICAM-1 molecule, thereby enhancing said ICAM-1/LFA-1 interaction.
Parent Case Info
This application is a division of application Ser. No. 08/229,513 filed Apr. 19, 1994 which application now abandoned.
US Referenced Citations (1)
Number |
Name |
Date |
Kind |
5340800 |
Lin et al. |
Aug 1994 |
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Foreign Referenced Citations (1)
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Non-Patent Literature Citations (2)
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Stanley et al. EMBO J. 13, 1790-1798, 1994. |
Divisions (1)
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Number |
Date |
Country |
Parent |
229513 |
Apr 1994 |
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