Smoothened polypeptides and methods of use

Abstract

Disclosed is an isolated or purified polypeptide or peptidomimetic comprising an amino acid sequence of a portion of a Smoothened (SMO) protein, wherein the portion comprises an amino acid sequence of any of the intracellular loops of the SMO protein, a functional fragment thereof, or a functional variant of either the portion or the functional fragment, wherein the functional fragment comprises at least 7 contiguous amino acids of the intracellular loops, and wherein the functional fragment or functional variant inhibits proliferation of a diseased cell, or a fatty acid derivative thereof. Related conjugates, nucleic acids, recombinant expression vectors, host cells, and pharmaceutical compositions are further provided. Methods of inhibiting proliferation of a diseased cell, treating or preventing cancer, treating a neoplasm or psoriasis, and inhibiting the expression of genes involved in the Hedgehog signaling pathway, thereby inhibiting the Hedgehog signaling pathway, are furthermore provided by the invention.

1 - 32 . (canceled) 
     
     
         33 . A method of inhibiting proliferation of a diseased cell, the method comprising contacting the diseased cell with a pharmaceutical composition comprising
 (i) an isolated or purified polypeptide, peptidomimetic, or fatty acid derivative of the polypeptide or peptidomimetic comprising an amino acid sequence comprising SEQ ID NO: 3, a functional fragment thereof, or a functional variant of either SEQ ID NO: 3 or the functional fragment, wherein the functional variant comprises an amino acid sequence (a) which has at least 90% sequence identity with SEQ ID NO: 3 or (b) which is a retroinverso analogue of SEQ ID NO: 3 or a functional fragment thereof, wherein the polypeptide or peptidomimetic is less than or equal to 50 amino acids in length, wherein the functional fragment comprises at least 7 contiguous amino acids of SEQ ID NO: 3, and wherein the functional fragment or functional variant inhibits proliferation of a diseased cell,   (ii) optionally a lipid, and a   (iii) pharmaceutically acceptable carrier in an amount effective to inhibit proliferation of the diseased cell.   
     
     
         34 . The method of  claim 33 , wherein the diseased cell is in a host. 
     
     
         35 . The method of  claim 34 , wherein the host is a mammal. 
     
     
         36 . The method of  claim 35 , wherein the mammal is a human. 
     
     
         37 . The method of  claim 33 , wherein the method treats or prevents a cancer of the host. 
     
     
         38 . The method of  claim 37 , wherein the cancer is breast cancer, prostate cancer, ovarian cancer, stomach cancer, colon cancer, liver cancer, melanoma, basal cell carcinoma, rhabdomyosarcoma, a medulloblastoma, pancreatic cancer, lung cancer, thyroid cancer, a myeloma, a lymphoma, a glioma, or a sarcoma. 
     
     
         39 . The method of  claim 38 , wherein the stomach cancer is a gastric adenocarcinoma. 
     
     
         40 . The method of  claim 33 , wherein the method treats or prevents a neoplasm of the host. 
     
     
         41 . The method of  claim 33 , wherein the pharmaceutical composition is topically administered to the host. 
     
     
         42 . The method of  claim 33 , wherein the pharmaceutical composition is intratumorally administered to the host. 
     
     
         43 . A method of treating or preventing cancer, psoriasis, or a neoplasm in a host, the method comprising administering to the host a pharmaceutical composition comprising
 (i) an isolated or purified polypeptide, peptidomimetic, or fatty acid derivative of the polypeptide or peptidomimetic comprising an amino acid sequence comprising SEQ ID NO: 3, a functional fragment thereof, or a functional variant of either SEQ ID NO: 3 or the functional fragment, wherein the functional variant comprises an amino acid sequence (a) which has at least 90% sequence identity with SEQ ID NO: 3 or (b) which is a retroinverso analogue of SEQ ID NO: 3 or a functional fragment thereof, wherein the polypeptide or peptidomimetic is less than or equal to 50 amino acids in length, wherein the functional fragment comprises at least 7 contiguous amino acids of SEQ ID NO: 3, and wherein the functional fragment or functional variant inhibits proliferation of a diseased cell,   (ii) optionally a lipid, and a   (iii) pharmaceutically acceptable carrier in an amount effective to treat or prevent the cancer, psoriasis, or a neoplasm.   
     
     
         44 - 45 . (canceled) 
     
     
         46 . A method of inhibiting the expression of a gene selected from the group consisting of Gli-1, Gli-2, Gli-3, Ptch, Shh, Smo, NES, and a combination thereof, in a diseased cell, the method comprising contacting the diseased cell with a pharmaceutical composition comprising
 (i) an isolated or purified polypeptide, peptidomimetic, or fatty acid derivative of the polypeptide or peptidomimetic comprising an amino acid sequence comprising SEQ ID NO: 3, a functional fragment thereof, or a functional variant of either SEQ ID NO: 3 or the functional fragment, wherein the functional variant comprises an amino acid sequence (a) which has at least 90% sequence identity with SEQ ID NO: 3 or (b) which is a retroinverso analogue of SEQ ID NO: 3 or a functional fragment thereof, wherein the polypeptide or peptidomimetic is less than or equal to 50 amino acids in length, wherein the functional fragment comprises at least 7 contiguous amino acids of SEQ ID NO: 3, and wherein the functional fragment or functional variant inhibits proliferation of a diseased cell,   (ii) optionally a lipid, and a   (iii) pharmaceutically acceptable carrier in an amount effective to inhibit the expression of the gene.   
     
     
         47 . A method of inhibiting the Hedgehog signal transduction pathway in a diseased cell, the method comprising contacting the diseased cell with a pharmaceutical composition comprising
 (i) an isolated or purified polypeptide, peptidomimetic, or fatty acid derivative of the polypeptide or peptidomimetic comprising an amino acid sequence comprising SEQ ID NO: 3, a functional fragment thereof, or a functional variant of either SEQ ID NO: 3 or the functional fragment, wherein the functional variant comprises an amino acid sequence (a) which has at least 90% sequence identity with SEQ ID NO: 3 or (b) which is a retroinverso analogue of SEQ ID NO: 3 or a functional fragment thereof, wherein the polypeptide or peptidomimetic is less than or equal to 50 amino acids in length, wherein the functional fragment comprises at least 7 contiguous amino acids of SEQ ID NO: 3, and wherein the functional fragment or functional variant inhibits proliferation of a diseased cell,   (ii) optionally a lipid, and a   (iii) pharmaceutically acceptable carrier in an amount effective to inhibit the Hedgehog signal transduction pathway.   
     
     
         48 - 50 . (canceled) 
     
     
         51 . The method of  claim 33 , wherein the polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 17 to 33. 
     
     
         52 . The method of  claim 33 , wherein the functional variant comprises the amino acid sequence of SEQ ID NO: 23 with one amino acid substitution at any of positions 1-7,9,11, and 12 of SEQ ID NO: 23 in which the amino acid at the position is substituted with Ala. 
     
     
         53 . The method of  claim 33 , wherein the functional variant comprises a retroinverso analogue of SEQ ID NO: 23, 26, or 33. 
     
     
         54 . The method of  claim 33 , wherein the functional variant comprises the amino acid sequence of any of SEQ ID NOs: 34 to 37, 68, 84, and 92-94. 
     
     
         55 . The method of  claim 33 , wherein the functional variant comprises the amino acid sequence of any of SEQ ID NOs: 38-59. 
     
     
         56 . The method of  claim 33 , wherein the fatty acid derivative comprises a fatty acid molecule at the amino (N-) terminus, the carboxyl (C-) terminus, or both the N- and C-termini, the fatty acid molecule optionally containing at least one amino group. 
     
     
         57 . The method of  claim 33 , wherein the functional variant comprises the amino acid sequence of SEQ ID NO: 68 (all D-amino acids), the functional variant is optionally amidated, and the amino acid at position 1 of the functional variant is optionally acetylated.