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Title: WO9110141A1: METHOD TO DETECT BONE AND OTHER CONNECTIVE TISSUE DISORDERS IN HUMANS AND ANIMALS
[ Derwent Title ]


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WO World Intellectual Property Organization (WIPO)
A1 INTERNATIONAL APPLICATION PUBLISHED WITH INTERNATIONAL SEARCH REPORT i

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43 pages

 
Inventor: ROBINS, Simon, Peter;

Assignee: THE ROWETT RESEARCH INSTITUTE
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Published / Filed: 1991-07-11 / 1990-12-28

Application Number: WO1990GB0002030

IPC Code: Advanced: A61K 39/395; C07K 16/00; C07K 16/18; C07K 16/44; C12P 21/08; G01N 33/50; G01N 33/53; G01N 33/577; G01N 33/68; G01N 33/70;
IPC-7: C12P 21/08; G01N 33/577; G01N 33/68;

ECLA Code: C07K16/18; C07K16/44; G01N33/68R; S01N800/10D;

Priority Number:
1989-12-30  GB1989000029366

Abstract:     The invention is directed to methods to assess connective tissue, especially bone, metabolism in disease or to monitor therapy, which method comprises assessing the levels of native free collagen-derived crosslinks in biological fluids, especially urine. The method can be enhanced by concomitantly determining the levels of an indicator of bone formation in biological fluids of the same individual and assessing the differences between the degradation marker and the formation indicator. Antibodies which are specifically immunoreactive with forms of crosslinks which occur free in biological fluids are also disclosed. [French]

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Designated Country: AT AU BB BE BF BG BJ BR CA CF CG CH CM DE DK ES FI FR GA GB GR HU IT JP KP KR LK LU MC MG ML MR MW NL NO RO SD SE SN SU TD TG

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First Claim:
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    Claims 1. A method to diagnose the presence of a disorder associated with connective tissue metabolism abnormalities, which method comprises comparing the level of native free crosslinks derived from collagen in a biological fluid of a subject with the level of said crosslinks in normal subjects so as to diagnose a subject having an enhanced level of said crosslinks as showing the presence of said disorder.

Description
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+ METHOD TO DETECT BONE AND OTHER
    CONNECTIVE TISSUE DISORDERS IN HUMANS AND ANIMALS
+ Background Art The-association of collagen as a major structural material in a multiplicity of tissues, including bone, cartilage, skin, tendons, dentine and various soft tissues is well known. It is also knownthat the fiber structure of collagen is stabilized by crosslinking. The presence of the fluorescent pyridinium ring system as a non-reducible crosslink in collagen was reported by Fujimoto, D., et al., J Biochem (1978) 83:863- 867. The Fujimoto paper reported isolation of a fluorescent peptide from pronase digestion of bovine Achilles tendon collagen. The isolated hydrolyzed pyridinoline (Pyd) was thought to contain three residues of hydroxylysine and it was recognized that, prior to hydrolysis, peptide fragments were attached to the pyridinoline moiety. Further work on characterization was conducted by Gunja-Smith, Z., et al., Biochem J (1981) 197:759-762-, using hydrolyzed urine, and advantage was taken of the presence of the pyridinoline in urine by Robins, S.P., Biochem J (1982) 207:617-620, who linked pyridinoline obtained from hydrolyzed urine to a carrier to raise antibodies. The antibodies were then employed in an immunoassay to determine the concentration of pyridinoline in hydrolyzed urine. The procedure was stated by Robins as useful to provide an index of the degradation of certain forms of mature collagen by analysis of physiological fluids.
    In all of the foregoing, hydrolyzates were employed to obtain total- pyridinoline since much of the crosslink retained peptide extensions of the hydroxylysyl residues responsible for its formation. Thus, in order to obtain a homogenous preparation containing the pyridinium ring, a preliminary hydrolysis step was necessary. By 1982, it was established that there were two pathways of crosslink formation depending on whether lysine or hydroxylysine residues were present in the telopeptides from which these crosslinks were derived (Robins, S.P., in "Collagen in Health and Disease" (1982) Weiss, J.B., et al., eds., pages 160-178, Churchill Livingstone, Edinburgh). This was stated to result in a specificity of crosslinking whereby in soft tissues, such as skin, reducible aldimine linkages are formed from oxidized lysyl residues, whereas-in cartilage and bone these bonds, initially formed from hydroxylysine aldehydes, undergo a spontaneous rearrangement to more stable oxoimine crosslinks. These bonds undergo further reaction to form 3-hydroxy-pyridinium crosslinks. The stable crosslinking pyridinoline analog involving lysine rather than hydroxylysine in the helix portion was identified and quantified by Ogawa, T., et al., Biochem Biophvs Res Commune (1982) 107:1251-1257; Eyre, D.R., et al., Anal Biochem (1984) 137:380-388, and designated deoxypyridinoline (Dpd). This material was then believed to be restricted to bone collagen, although amounts vary between species.
+ Brief Description of the Drawings Figure 1 shows a chromatographic trace of pyridinoline obtained from an acid hydrolyzate superimposed on a trace of the pyridinoline obtained without hydrolysis from urine. The figure further compares the elution pattern as determined by fluorescence with the elution pattern as determined by reaction with antipyridinoline antibody prepared from hydrolyzate.
    Modes of Carrying Out the Invention The invention provides an improvement over the presently available methods to diagnose bone disorders or other diseases characterized by abnormalities in collagen metabolism. The invention method utilizes variations in the levels of collagen-derived pyridinium crosslinks in biological fluid as an index of these abnormalities.

       
Forward References: Go to Result Set: Forward references (9)
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Patent  Pub.Date  Inventor Assignee   Title
Buy PDF- 5pp US6033918  2000-03-07 Hatch; Robert P.  Bayer Corporation Method and device for the detection of analyte in a fluid sample
Buy PDF- 20pp US5972623  1999-10-26 Krane; Stephen M.  Metra Biosystems, Inc. Collagen-peptide assay method
Buy PDF- 14pp US5756361  1998-05-26 Winterbottom; Neil  Metra Biosystems, Inc. Screening method for periodontal disease
Buy PDF- 18pp US5736344  1998-04-07 Kung; Viola T.  Metra Biosystems, Inc. Serum pyridinium crosslinks assay
Buy PDF- 11pp US5661039  1997-08-26 Kung; Viola T.  Metra Biosystems, Inc. Perspiration assay for bone resorption
Buy PDF- 22pp US5620861  1997-04-15 Cerelli; Mary J.  Metra Biosystems, Inc. Method and kit for pyridinium crosslink assay
Buy PDF- 5pp US5545534  1996-08-13 Akita; Mikio  Hoechst Japan Limited Method for screening for osteoporosis
Buy PDF- 16pp US5527715  1996-06-18 Kung; Viola T.  Metra Biosystems, Inc. Method and kit for pyridinoline assay
Buy PDF- 6pp US5494802  1996-02-27 Watanabe; Hiroshi  Hoechst Japan Limited Tissue antigen and diagnostic method for human osteoporosis using the same
       
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