Structure-activity relationship studies for the peptide portion of the bladder epithelial cell antiproliferative factor from interstitial cystitis patients.
We performed comprehensive structure-activity relationship (SAR) studies on the peptide portion of antiproliferative factor (APF), a sialylated frizzled-8 related glycopeptide that inhibits normal bladder epithelial and urothelial carcinoma cell proliferation. Glycopeptide derivatives were synthesized by solid-phase methods using standard Fmoc chemistry and purified by RP-HPLC; all intermediate and final products were verified by HPLC-MS and NMR analyses. Antiproliferative activity of each derivative was determined by inhibition of (3)H-thymidine incorporation in primary normal human bladder epithelial cells. Structural components of the peptide segment of APF that proved to be important for biological activity included the presence of at least eight of the nine N-terminal amino acids, a negative charge in the C-terminal amino acid, a free amino group at the N-terminus, maintenance of a specific amino acid sequence in the C-terminal tail, and trans conformation for the peptide bonds. These data provide critical guidelines for optimization of structure in design of APF analogues as potential therapeutic agents.
Kaczmarek
Piotr
P
Structural Biophysics Laboratory, Center for Cancer Research, National Cancer Institute at Frederick, Frederick, Maryland 21702, USA.
Keay
Susan K
SK
Tocci
Gillian M
GM
Koch
Kristopher R
KR
Zhang
Chen-Ou
CO
Barchi
Joseph J
JJ
Grkovic
David
D
Guo
Li
L
Michejda
Christopher J
CJ
D000293:Adolescent; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D049109:Cell Proliferation / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D018856:Cystitis, Interstitial / Q000652:urine*; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000737:chemistry / Q000187:drug effects*; D006020:Glycopeptides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D006023:Glycoproteins / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D006131:Growth Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008968:Molecular Conformation; D013237:Stereoisomerism; D013329:Structure-Activity Relationship; D001743:Urinary Bladder / Q000737:chemistry / Q000166:cytology*
D000596:Amino Acids; D006020:Glycopeptides; D006023:Glycoproteins; D006131:Growth Inhibitors; D036341:Intercellular Signaling Peptides and Proteins; C490708:antiproliferative factor APF, human
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