US005652112A United States Patent 19 11 Patent Number: 5,652,112 Eyre 45 Date of Patent: Jul. 29, 1997

54 ASSAY FORTYPE I CARBOXY. Baldwin, et al., "Structure of cDNA clones coding for TERMINAL TELOPEPTDEANALYTES human type II procollagen; the a1(II) chain is more similar to the a1 (I) chain than two other a chains of fibrillar (75) Inventor: David R. Eyre, Mercer Island, Wash. .” Journal, 262:521-528 (1989). Ala-Kokko, et al., "Structure of cDNA clones coding for the 73) Assignee: Washington Research Foundation, entire preproal(II) chain of human type III procollagen: Seattle, Wash. Differences in protein structure from type I procollagen and conservation of codon preferences,” Biochemistry Journal, 21 Appl. No.: 771,219 260:509-516 (1989). Seibel, “Komponentender extrazllularen Gewebematrix als 22 Filed: Dec. 20, 1996 potentielle Marker des Bindegewebs-, Knorbel-und Knochenmetabolismus bei Erkrankungen des Bewegung Related U.S. Application Data sapparates.” Zeitschrift fur Rheumatologie, 48:6-18 (1989). 63 Continuation of Ser. No. 537,502, Oct. 2, 1995, which is a Dodge and Poole, "Immunlohistochemical Detection and continuation of Ser. No. 226,070, Apr. 11, 1994, Pat. No. Immunochemical Analysis of Type II Collagen Degradation 5,455,179, which is a continuation of Ser. No. 823,270, Jan. in Human Normal, Rheumatoid, and Osteoarthritic Articular 16, 1992, Pat No. 5,532,169, which is a division of Ser. No. Cartilages and in Explants of Bovine Articular Cartilage 444,881, Dec. 1, 1989, Pat No. 5,140,103, which is a continuation-in-part of Ser. No. 118,234, Nov. 6, 1987, Pat. Cultured with Interleukin 1.” Journal of Clinical Investiga No. 4973,666. w tion, 83:647-661 (1989). Niemela, "Radioimmunoassays for Type II Procollagen (51 int. Cl. ...mmm. GO1N 33/536 Amino-Terminal Peptides In Humans,” Clinical Chemistry, 52 U.S. C...... 435/7.1; 436/518; 436/536 31:1301-1304 (1985). 58 Field of Search ...... 435/7.1; 436/518, Sangiorgi, et al., “Isolation and partial characterization of 436/536 the entire human proa1 (TT) collagen gene.” Nucleic Acids Research, 13:2207-2225 (1985). 56 References Cited Loidi, et al., “Molecular cloning and carboxyl-propeptide analysis of human type ITI procollagen,” Nucleic Acids U.S. PATENT DOCUMENTS Research, 12:9383-9394 (1984). 3,600,132 8/1971 Goverde ...... 23/230 Wu and Eyre, "Identification of Hydroxypyridinium 4,094,646 6/1978 Stern et al...... 23/230 Cross-Linking Sites in Type II Collagen of Bovine Articular 4,312,853 1/1982 Timpl. Cartilage,” Biochemistry, 23:1850-1857 (1984). 4,371,374 2/1983 Cerami et al...... 23/230 Pierard, et al., "Radioimmunoassay for the Amino-Terminal 4504587 3/1985 Timplet al.. Sequences of Type III Procollagen in Human Body Fluids 4,731,326 3/1988 Thompson et al...... 435/7

Measuring Fragmented Precursor Sequences,” Analytical 4,774,227 9/1988 Piez et al...... 54/21 Biochemistry, 141:127-136 (1984). 5,455,179 10/1995 Eyre ...... 436/536 Rohde, et al., “Radioimmunoassay for type III procollagen FOREIGN PATENT DOCUMENTS peptide and its application to human liver disease," Euro pean Journal of Clinical Investigation, 9:451-459 (1979). 128 041 6/1983 European Pat. Off...... CO7G 7/OO 289 314 4/1988 European Pat. Off...... A61K 37/36 Russell, R.G.G., et al., “Biochemical Markers of Bone 0505 210 A2 9/1992 European Pat. Off.. Turnover in Paget's Disease.” Metab. Bone Dis. & Rel. Res., 0502 928 B1 1/1996 European Pat. Off.. 4 & 5:255-262 (1981). 2205643 5/1987 United Kingdom. Drinkwater, B.L., et al., “Bone Mineral Density After WO 90/0898O 11/1988 WIPO Resumption of Menses in Amenorrheic Athletes,” JAMA, WO 89/0449 5/1989 WIPO 256:380-382 (1986). WO 89/12824 12A1989 WIPO WO 90/04417 5/1990 WIPO (List continued on next page.) WO 92/21698 12/1992 WIPO. Primary Examiner-James Ketter OTHER PUBLICATIONS Attorney, Agent, or Firm-Christensen O'Connor Johnson & Kindness PLLC Hanson et al., "A specific immunoassay for monitoring human bone resorption: quantitation of type I collagen 57 ABSTRACT cross-linked N-telopeptides in urine,” Journal of Bone and A method of determining collagen degradation in vivo, Mineral Research, 7:1251-1258 (1992). comprising quantitating the concentration of a peptide in a Sakura, et al., “Photolysis of pyridinoline, a cross-linking body fluid, the peptide being a C-terminal type II collagen of collagen, by ultraviolet light,” Can. J. Bio telopeptide containing a hydroxylysyl pyridinoline cross chem, 60:525-529 (1982). link or a type III collagen telopeptide containing a hydroxy Black, et al., “Urinary excretion of the hydroxypyridinium lysyl pyridinoline cross-link. The method includes immu cross links of collagen in patients with rheumatoid arthritis.” nometric assays, fluorometric assays, and electrochemical Annals of the Rhemuatic Diseases, 48:641-644 (1989). titrations for quantitation. The structures of specific peptides Seibel, et al., “Urinary Hydroxy-pyridinium Crosslinks Pro having cross-links and kits for quantitating these peptides in vide Indices of Cartilage and Bone Involvement in Arthritic a body fluid are described. Diseases,” the Journal of Rhemuatology, 16:964-970 (1989). 2 Claims, 10 Drawing Sheets 5,652,112 Page 2

OTHER PUBLICATIONS Eyre, D.R., et al., "Quantitation of Hydroxypyridinium Crosslinks in Collagen by High-Performance Liquid Chro Drinkwater, B.L., et al., "Bone Mineral Content of Amen matography,” Analytical Biochemistry, 137:380-388 orrheic and Eumenorrheic Athletes,” the New England Jour (1984). nal of Medicine, 311:5; 277-281 (1984). Robins, S.R., et al., “Measurement of the cross linking Fujimoto, D., "Evidence for Natural Existence of Pyridino compound, pyridinoline, in urine as an index of collagen line Crosslink in Collagen.” Biochemical and Biophysical degradation in joint disease.” Annals of the Rheumatic Research Communications, 93:948-953 (1980). Diseases, 45:969–973 (1986). Yamauchi, M. et al., “A Comparative Study of the Distri Bernard, M.P., et al., “Nucleotide Sequences of Comple bution of the Stable Crosslink, Pyridinoline, in Bone Co mentary Deoxyribonucleic Acids for the Proa1 Chain of lagens from Normal, Osteoblastoma, and Vitamin D-Defi Human Type I Procollagen. 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Fujimoto, D., et al., “Pyridinoline, A Non-Reducible Whittle, et al., "Biochemical investigation of possible Crosslink of Collagen. Quantitative Determination, Distri lesions in human aorta that predispose to dissecting aneu bution, and Isolation of a Crosslinked Peptide,” Chemical rysms: pyridinoline crosslinks.” Cardiovascular Research Abstracts 89(11):148, (Sep. 11, 1978). 21:161-168 (1987). Black, D., et al., “Quantitative Analysis of the Pyridinium Gunja-Smith, Z., "An Enzyme-Linked Immunosorbent Crosslinks of Collagen in Urine Using Ion-Paired Assay to Quantitate the Elastin Crosslink in Reversed-Phase High-Performance Liquid Chromatogra Tissue and Urine Samples,” Analytical Biochemistry phy,” Chemical Abstracts 108(21):354, (May 23, 1988). 147:258-264 (1985). 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Eyre, et al., "Identification of urinary peptides derived from Delmas, Pierre D., "Biochemical Markers of Bone Turnover cross-linking sites in bone collagen in Paget's Disease,” for the Clinical assessment of Metabolic Bone Disease.” Journal of Bone and Mineral Research, 3, Suppl. 1 (Jun. Endocrinology and Metabolism Clinics of North America, 1988). 19(1) (Mar. 1990). Eyre, et al., "Reducible Crosslinks in Hydroxylysine-Defi Uebelhart, et al., “Urinary excretion of pyridinium cient Collagens of a Heritable Disorder of Connective crosslinks: a new marker of bone resorption in metabolic Tissue,” Proc. Nat. Acad. Sci. USA 69(9):2594-2598 (Sep. bone disease” Bone and Mineral, 8:87-96 (1990). 1972). 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Current Clinical and Fundamental Prob Gunja-Smith & Boucek, “ in human urine.” lems, Paris, France: Ciba-Geigy, 1984, pp. 117-122. Biochem. J., 193:915-918, 1981. U.S. Patent Jul. 29, 1997 Sheet 1 of 10 5,652,112

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