Human respiratory syncytial virus
AssigneeWhitehead Institute for Biomedical Research
InventorsXun Zhao, Mona Singh, Vladimir Malashkevich, Peter S. Kim
Life of this patent
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Abstract
A method of identifying a drug that inhibits the RSV membrane fusion machinery, such as the HRSV membrane fusion machinery; drugs that bind the N-helix coiled-coil of HRSV, particularly the N-helix coiled-coil pocket, and, thus, inhibit HRSV infection of cells, such as human cells; methods of inhibiting (partially or totally) HRSV infection of an individual; anitbodies that bind to the N-helix coiled-coil region of HRSV F1; and pharmaceutical compositions comprising HRSV infection neutralizing antibodies.
Claims37
A method of identifying a drug that inhibits the RSV membrane fusion machinery by inhibiting interactions between the N57 peptide trimer and the C45 peptide trimer of RSV F1, comprising:
- (a) combining RSV F1 N57 peptide trimer, RSV F1 C45 peptide trimer and a drug to be assessed for its ability to inhibit interaction between the two trimers, to produce a combination;
- (b) maintaining the combination under conditions appropriate for interactions to occur between N57 peptide trimers and C45 peptide trimers;
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- (c) assessing whether interactions occurred between N57 peptide trimers and C45 peptide trimers, wherein if interactions between the N57 peptide trimer and the C45 peptide trimer did not occur in the presence of the drug or occurred to a lesser extent in the presence of the drug than in its absence, the drug is a drug that inhibits the RSV membrane fusion machinery.
A method of producing a drug which inhibits interaction of two components of the core of fusion-active HRSV envelope F1, wherein the two components are referred to as N57 peptide trimer and C45 peptide trimer, respectively, comprising identifying a compound or designing a compound which fits into a hydrophobic groove formed by two N57 peptide chains which form the semipocket, wherein the semipocket is lined by K191, L195 and Y198 from a first N57 chain and K196, D200 and L204 from a second N57 chain.
A method of producing a drug which inhibits interaction of N57 peptide trimer with C45 peptide trimer, wherein N57 peptide trimer and C45 peptide trimer comprise the core of fusion-active HRSV envelope F1, comprising identifying a compound or designing a compound which fits into a hydrophobic groove formed by two N57 peptide chains which form a semipocket, wherein the semipocket is lined by K191, L195 and Y198 from a first N57 chain and K196, D200 and L204 from a second N57 chain.
A compound which inhibits interaction of N57 peptide trimer of the α-helical domain of HRSV F1 which is the core or fusion active F1 with C45 peptide trimer of the α-helical domain.
The compound of claim 8 wherein the compound fits into a hydrophobic groove formed by two N57 peptide chains which form the semipocket, wherein the semipocket is lined by K191, L195 and Y198 from a first N57 chain and K196, D200 and L204 from a second N57 chain.
A peptide which comprises a soluble, trimeric form of a coiled coil and a sufficient portion of the N-peptide region of HRSV F1 to comprise the amino acid residues which form a semi-pocket of the N-helix coiled coil of HRSV F1.
A method of identifying a drug that interferes with the formation of a complex between C45 peptide and N57 peptide, comprising:
- (a) combining a candidate drug to be assessed for its ability to interfere with formation of a complex between C45 peptide and N57 peptide, under conditions appropriate for formation of a complex between C45 peptide and N57 peptide, thereby forming a test sample;
- and
- (b) determining whether formation of a complex between C45 peptide and N57 peptide is inhibited, wherein if formation of the complex is inhibited, the candidate drug is a drug that interferes with formation of the complex whereby a drug that interferes with formation of the complex is identified.
A method of eliciting an immune response in an individual, comprising introducing into the individual a peptide comprising a trimeric form of a coiled-coil region of a protein and a sufficient portion of the N-peptide region of RSV F1 to comprise the amino acid residues which form part or all of the N-helix coiled-coil of RSV F1 and the peptide is present in a pharmaceutically acceptable carrier.
A method of identifying a compound or molecule which binds the N-helix coiled-coil semi-pocket of HRSV F1 envelope protein, wherein the compound or molecule to be assessed is referred to as a candidate inhibitor, comprising:
- (a) combining a D-peptide which binds the N-helix coiled-coil cavity, a fusion protein which is a soluble model which presents the N-helix coiled-coil cavity and a candidate inhibitor, under conditions appropriate for binding of the D-peptide to the N-helix coiled-coil cavity, thereby producing a test sample;
- (b) determining the extent to which binding occurs of the D-peptide to the N-helix coiled-coil semi-pocket in the test sample;
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- (c) comparing the extent of binding determined in the N-helix coiled-coil semi-pocket in a control sample, wherein the control sample is the same as the test sample except that the control sample does not include the candidate inhibitor and is maintained under the same conditions appropriate for binding of the D-peptide to the N-helix coiled-coil semi-pocket as is the test sample, wherein if the extent of binding in the test sample is less than the extent of binding in the control sample, the candidate inhibitor is a compound or molecule which binds the N-helix coiled-coil semi-pocket of RSV F1 envelope protein.
A method of eliciting an immune response in an individual, comprising introducing into the individual a fusion protein comprising a soluble, trimeric form of a coiled-coil and a sufficient portion of the N-peptide region of HRSV F1, to comprise the amino acid residues which form the pocket of the N-helix coiled-coil of HRSV F1, wherein the fusion protein is present in a pharmaceutically acceptable carrier.
A method of identifying a drug that binds the N-helix coiled-coil semi-pocket of HRSV F1 comprising:
- (a) combining:
- (1) a candidate drug to be assessed for its ability to bind the N-helix coiled-coil semi-pocket of HRSV F1 and;
- (2) a fusion protein which comprises a trimeric version of the coiled-coil region of a protein and a sufficient portion of the N-peptide of HRSV F1 to include the HRSV F1 semi-pocket, under conditions appropriate for presentation of the HRSV F1 semi-pocket for binding by a drug;
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- (b) determining whether the candidate drug binds the HRSV F1 semi-pocket, wherein if binding occurs, the candidate drug is a drug which binds the N-helix coiled-coil semi-pocket of HRSV F1.
A method of identifying a drug that binds the N-helix coiled-coil semi-pocket of HRSV F1 comprising:
- (a) combining:
- (1) a soluble model that presents the N-helix coiled-coil semi-pocket of HRSV F1 in such a manner that it is available for binding by a drug and (2) a candidate drug, which is to be assessed for its ability to bind the N-helix coiled-coil semi-pocket;
- and
- (b) determining whether the candidate drug binds the N-helix coiled coil semi-pocket of the soluble model, wherein if binding occurs, the candidate drug is a drug which binds the N-helix coiled-coil semi-pocket of HRSV F1.
A method of producing a drug that binds the N-helix coiled-coil semi-pocket of HRSV F1 and inhibits HRSV entry into cells, comprising:
- (a) combining (1) a candidate drug to be assessed for its ability to bind the N-helix coiled-coil semi-pocket of HRSV F1 and inhibit HRSV entry into cells and (2) a fusion protein which comprises a trimeric version of the coiled-coil region of a protein and a sufficient portion of the N-peptide of HRSV F1 to include the HRSV F1 semi-pocket, under conditions appropriate for presentation of the HRSV F1 semi-pocket for binding by a drug;
- (b) determining whether the candidate drug binds the HRSV F1 semi-pocket, wherein if binding of the candidate drug to the N-helix coiled-coil semi-pocket of HRSV F1 occurs, the candidate drug is a drug which binds the N-helix coiled-coil semi-pocket of HRSV F1, whereby a drug which binds the N-helix coiled-coil semi-pocket of HRSV F1 is produced;
- and
- (c) assessing the ability of the drug produced in (b) to inhibit HRSV entry into cells, wherein if the drug inhibits HRSV entry into cells, it is a drug which binds the N-helix coiled-coil semi-pocket of HRSV F1 and inhibits HRSV entry into cells.
A method of producing a soluble model of the N-helix coiled-coil semi-pocket of HRSV F1, comprising producing a fusion protein comprising:
- (a) a soluble, trimeric form of a coiled-coil and (b) a sufficient portion of the N-peptide region of HRSV F1 to comprise the amino acid residues which form the pocket of the N-helix coiled-coil of HRSV F1.
A method of producing a drug that binds the N-helix coiled-coil semi-pocket of HRSV F1 comprising:
- (a) producing or obtaining a soluble model of the N-helix coiled-coil semi-pocket of HRSV F1;
- (b) combining:
- (1) a candidate drug to be assessed for its ability to bind the N-helix coiled-coil semi-pocket of HRSV F1 and (2) the soluble model of the N-helix coiled-coil semi-pocket of HRSV F1;
- and
- (c) determining whether the candidate drug binds the N-helix coiled-coil semi-pocket of HRSV F1, wherein if the candidate drug binds the N-helix coiled-coil semi-pocket of HRSV F1, the candidate drug is a drug which binds the N-helix coiled-coil semi-pocket of HRSV F1, whereby a drug which binds the N-helix coiled-coil semi-pocket of HRSV F1 is produced.
A method of producing a drug that binds the N-helix coiled-coil semi-pocket of HRSV F1 and inhibits its entry into cells, comprising:
- (a) producing or obtaining a soluble model of the N-helix coiled-coil semi-pocket of HRSV F1;
- (b) combining:
- (1) a candidate drug to be assessed for its ability to bind the N-helix coiled-coil semi-pocket of HRSV F1 and (2) the soluble model of the N-helix coiled-coil semi-pocket of HRSV F1;
- (c) determining whether the candidate drug binds the N-helix coiled-coil semi-pocket of HRSV F1, wherein if the candidate drug binds the N-helix coiled-coil semi-pocket of HIV gp41, the candidate drug is a drug which binds the N-helix coiled-coil semi-pocket of HRSV F1, whereby a drug which binds the N-helix coiled-coil semi-pocket of HRSV F1 is produced and;
- (d) assessing the ability of the drug produced in (c) to inhibit HRSV entry into cells, wherein if the drug inhibits HRSV entry into cells, it is a drug which binds the N-helix coiled-coil semi-pocket of HRSV F1 and inhibits HRSV entry into cells.
An isolated antibody that binds to a peptide, wherein the peptide comprises a soluble, trimeric form of a coiled-coil and a sufficient portion of the N-peptide region of HRSV F1 to comprise the amino acid residues that form part or all of the N-helix coiled-coil of HRSV F1 and the antibody binds to the sufficient portion of the peptide.
An isolated antibody that binds the coiled-coil region of HRSV F1 as presented by a peptide which presents the N-helix coiled-coil region of HRSV F1 as a properly folded trimer.
Dependencies are inferred from the claim text.