Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Peptide containing doai
Patent
1996-01-22
1999-06-01
Prouty, Rebecca E.
Drug, bio-affecting and body treating compositions
Designated organic active ingredient containing
Peptide containing doai
4353201, 435 691, 4352523, 43525411, 4352542, 536 231, 536 235, 536 2431, 530412, A61K 3800
Patent
active
059088273
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates to a new protein named Component B. In particular the invention relates to a new protein obtainable from urine, its preparation from urine, its production by recombinant DNA techniques using genomic DNA or cDNA encoding said new protein, as well as pharmaceutical compositions containing it and its use in therapy.
SUMMARY OF THE INVENTION
A new protein was isolated during the extraction and purification process of urine derivatives. This protein shows a polypeptide nature and relatively low molecular weight. When human urine is treated with adsorbing materials, as kaolin, and then undergoes filtration, ion exchange chromatography and high resolution chromatography, preferably according to the process hereafter described, after lyophilisation a compound is obtained as amorphous white powder, moving as an a single peak in high pressure reversed phase liquid chromatography (HPLC-RP) and having a molecular weight of about 9 KDa when analysed by electrophoresis on a polyacrylamide gel in the presence of sodium dodecyl sulphate (SDS-Page) under reducing conditions. This protein was named, and is referred to hereinafter as, Component B.
Component B is more specifically characterised through the amino acid sequence reported as SEQ ID NO: 1.
The present invention makes therefore available a new protein, named Component B, obtainable through a process comprising the isolation of a raw fraction of the compound itself from a dialysed concentrate of urine after treatment with an adsorbing agent and its purification by ion exchange chromatography and high resolution chromatography as described hereafter.
Preferably the protein according to the present invention is extracted from human urine because of the high amount available. This technique is thus useful for industrial production. The present invention refers particularly to a polypeptide comprising the SEQ ID NO: 1, its salts, functional derivatives, precursors and active fractions as well as its active mutants, i.e. other proteins or polypeptides wherein one or more amino acids of the structure were eliminated or substituted by other amino acids or one or more amino acids were added to that sequence in order to obtain polypeptides or proteins having the same activity of Component B and comprises also the corresponding fusion proteins i.e. polypeptides comprising Component B or a mutation thereof fused with another protein and having a longer lasting half-life in body fluids. Component B can therefore be fused with another protein such as, for example, an immunoglobulin.
The definition "salts" as used herein refers both to salts of the carboxyl-groups and to the salts of the amino functions of the compound obtainable through known methods.
The salts of the carboxyl-groups comprise inorganic salts as, for example, sodium, potassium, calcium salts and salts with organic bases such as those formed with an amine as triethanolamine, arginine or lysine. The salts of the amino groups comprise, for example salts with inorganic acids such as hydrochloric acid and with organic acids such as acetic acid. The definition "functional derivatives" as herein used refers to derivatives which can be prepared from the functional groups present on the lateral chains of the amino acid moieties or on the terminal N- or C-groups according to known methods and are comprised in the invention when they are pharmacetically acceptable i.e. when they do not destroy the protein activity or do not impart toxicity to the pharmaceutical compositions containing them.
Such derivatives include for example esters or aliphatic amides of the carboxyl-groups and N-acyl derivatives of free amino groups or O-acyl derivatives of free hydroxyl-groups and are formed with acyl-groups as for example alcanoyl- or aroyl-groups.
The "precursors" are compounds which are converted into the Component B in the human or animal body. As "active fractions" of the protein the present invention refers to any fragment or precursor of the polypeptidic chain of the compou
REFERENCES:
patent: 5298604 (1994-03-01), Sloane
Accession No. R3O17 Cohen et al. (1993) Standard Protein; 131 AA.
Accession No. Q2719O Lee et al. (1991) Standard cDNA; 1414 BP.
Accession No. Q1O572 Chang M. et al. (1991) Standard DNA; 1047 BP.
Sloane et al. (1986) Biochem. J., 234:355-362.
R. Ridge et al, "Partial N-Terminal Amino Acid Sequence of the Anti-Neoplastic Urinary Protein (ANUP) and the Anti-Tumor Effect of the N-Terminal Nonapeptide of the Unique Cytokine Present in Human Granulocytes", Cytokine, vol. 8, No. 1 (Jan.), 1996 pp. 1-5.
N. Sloane et al, "Studies on an Antineoplastic Fraction from Human Urine", Biochem. J. (1986) 234, pp. 355-362.
Clinica Chimica Acta, vol. 207 (1992), pp. 239-249. A. Bernard et al., "Human Urinary protein 1: Evidence for identity with Clara cell protein and occurence in respiratory tract and urogenital secretions."
Applied Research Systems ARS Holding N.V.
Longton Enrique D.
Prouty Rebecca E.
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