Three dimensional coordinates of HPTPbeta
View Patent ↗The crystal structures of catalytic domain of HPTPbeta, both ligand-bound and ligan-free are described. These structures are useful in computer aided drug design for identifying compounds that bind or activate HPTPbeta and thereby modulate angiogenesis mediated disorders or diseases.
1. A method of identifying a drug candidate compound for the treatment of an angiogenic mediate disorder, comprising:
a) employing the three-dimensional structural coordinates of the human protein tyrosine phosphatase beta (HPTPbeta) catalytic domain [SEQ ID NO:7] as set forth in FIGS. 202-252 to graphically image the HPTPbeta catalytic domain [SEQ ID NO: 7], and determining the binding mode of a compound within the catalytic domain;
b) selecting one or more compounds which have the best fit with the HPTPbeta catalytic domain [SEQ ID NO:7] as a set forth in FIGS. 202-252 wherein the compounds are computationally positioned at one or more areas of said imaged HPTPbeta catalytic domain [SEQ ID NO:7] and
c) assaying analyzing the ability of said drug candidate compound to bind or modulate HPTPbeta activity.
2. A method of identifying a drug candidate compound for the treatment of an angiogenic mediate disorder, comprising:
a) employing the three-dimensional structural coordinates of the HPTPbeta catalytic domain [SEQ ID NO:7] as set forth in FIGS. 7-102 to graphically image the HPTPbeta catalytic domain [SEQ ID NO:7], and determining the binding mode of a compound within the catalytic domain;
b) selecting one or more compounds which have the best fit with the HPTPbeta catalytic domain [SEQ ID NO:7] as set forth in FIGS. 7-102 wherein the compounds are computationally positioned positioning a drug candidate compound at one or more areas of said imaged HPTPbeta catalytic domain [SEQ ID NO:7]; and
c) assaying the ability of said drug candidate compound to bind or modulate HPTPbeta activity.
3. The method according to claim 1 wherein said drug candidate compound is positioned at at least amino acid residues 152, 74-77, 209-214, 244-253, 288-290, and 293 of [SEQ ID NO:7].
4. The method according to claim 2 wherein said drug candidate compound is positioned at at least amino acid residues 152, 74-77, 209-214, 244-253, 288-290 , and 293 of [SEQ ID NO:7].
5. The method according to claim 1 wherein said drug candidate compound is positioned at at least amino acid residues 76-80, 48-66, 284-292 and 212-214 of [SEQ ID NO:7].
6. The method according to claim 2 wherein said drug candidate compound is positioned at at least amino acid residues 76-80, 48-66, 284-292 and 212-214 of [SEQ ID NO:7].
7. The method according to claim 1 wherein said drug candidate compound is positioned at at least amino acid residues 69-76, 119-123, and 149-154 of [SEQ ID NO:7].
8. The method according to claim 2 , wherein said drug candidate compound is positioned at at least amino acid residues 69-76, 119-123, and 149-154 of [SEQ ID NO:7].
9. A method of identifying an inhibitor of an HPTPbeta molecule, comprising:
a) providing a crystal of HPTPbeta comprising the amino acid residues of SEQ ID NO:7 which either has space group P2 1 2 1 2 1 and unit cell parameters a=39 Å, b=72 Å, c=120 Å and α=β=γ=90°, or space group P2 1 with unit cell parameters of a=62 Å, b=70 Å, c=70 Å and α=90°, β=93°, γ=90°;
b) subjecting the crystal in (a) to X-ray diffraction and determining the three-dimensional structure coordinates of the HPTPbeta catalytic domain;
c) employing the three-dimensional structure coordinates from (b) to graphically display the three-dimensional structure and identifying the active binding site residues of Asn74 Asn75 Ile76, Leu77, Cys152, Pro209, Asp210, His211, Gly212, Va1213, Pro214, Cys244, Ser245, Ala246, Gly247, Va1248, Gly249, Arg250, Thr251, Gly252, Thr253, Gin288, Thr289, Glu290, and Tyr293;
d) using the atomic coordinates of amino acids Asn74 Asn75 Ile76, Leu77, Cys 152, Pro209, Asp210, His211, Gly212, Va1213, Pro214, Cys244, Ser245, Ala246, Gly247, Va1248, Gly249, Arg250, Thr251, Gly252, Thr253, Gin288, Thr289, Glu290, and Tyr293 as determined in c) to generate a three-dimensional structure of a molecule comprising an HPTPbeta active site binding pocket so that the root mean square deviation of conserved residue backbone atoms is less than 1.5 Å as compared to the three-dimensional coordinates in a-c);
e) employing the three-dimensional structure from d) to design or select a potential drug candidate; and
f) contacting the potential drug candidate with the molecule to determine the abilut of the potential inhibitor to associated with the molecule.
10. A method of identifying an inhibitor of an HPTPbeta molecule, comprising:
a) providing a crystal of HPTPbeta comprising the amino acid residues of SEQ ID NO:7 which either has space group P2 1 2 1 2 1 and unit cell parameters a=39 Å, b=72 Å, c=120 Å and α=β=γ=90°, or space group P2 1 with unit cell parameters of a=62 Å, b=70 Å, c=70 Å and α=90°, β=93°, γ=90°;
b) subjecting the crystal in (a) to X-ray diffraction and determining the three-dimensional structure coordinates of the HPTPbeta catalytic domain;
c) employing the three-dimensional structure coordinates from (b) to graphically display the three-dimensional structure and identifying the active binding site residues of Glu48, Glu49, Leu50, Lys51, Asp52, Va153, Gly54, Arg55, Asn56, Gln57, Ser58, Cys59, Asp60, Ile61, Ala62, Leu63, Leu64, Pro65, Glu66, Ile76, Leu77, Pro78, Tyr79, Asp80, Gly212, Va1213, Pro214, Va1284, His285, Met286, Va1287, Gln288, Thr289, Glu290, Cys291, and Gln292;
d) using the atomic coordinates of amino acids Glu48, Glu49, Leu50, Lys51, Asp52, Va153, Gly54, Arg55, Asn56, Gln57, Ser58, Cys59, Asp60, Ile61, Ala62, Leu63, Leu64, Pro65, Glu66, Ile76, Leu77, Pro78, Tyr79, Asp80, Gly212, Va1213, Pro214, Va1284, His285, Met286, Va1287, Gln288, Thr289, Glu290, Cys291, and Gln292 as determined in c) to generate a three-dimensional structure of a molecule comprising an HPTPbeta active site binding pocket so that the root mean square deviation of conserved residue backbone atoms is less than 1.5 Å as compared to the three-dimensional coordinates in a-c);
e) employing the three-dimensional structure from d) to design or select a potential drug candidate; and
f) containing the potential drug candidate with the molecule to determine the ability of the potential inhibitor to associate with the molecule.