Drug – bio-affecting and body treating compositions – Nonspecific immunoeffector – per se ; or nonspecific...
Reexamination Certificate
1999-12-03
2003-05-20
Nolan, Patrick J. (Department: 1644)
Drug, bio-affecting and body treating compositions
Nonspecific immunoeffector, per se ; or nonspecific...
C530S350000, C530S400000
Reexamination Certificate
active
06565858
ABSTRACT:
TECHNICAL FIELD
The present invention relates to a human ADAMTS-1 protein, a gene encoding the same, a pharmaceutical composition, and a method for immunologically analyzing the human ADAMTS-1 protein.
BACKGROUND ART
A mouse ADAMTS (A disintegrin and metalloproteinase with thrombospondin motifs)-1 gene has been cloned as a cDNA from a mouse colon cancer cell which induces cancer cachexia when transplanted to a mouse. The mouse ADAMTS-1 protein encoded by the gene is a unique protein containing a matrix metalloproteinase domain, a disintegrin domain, and three thrombospondin domains [J. Biol. Chem., 272, 556-562 (1997)]. The physiological functions of the mouse ADAMTS-1 protein has not been reported, but there have been reports of each individual functional domain contained therein.
For example, a snake venom disintegrin belongs to a family of proteins which are rich in cystein, and exhibit an anticoagulant activity [Semin. Hematol., 31, 289-300 (1994)].
Further, for example, an ADAM (A disintegrin and metalloproteinase) family is known as a protein family containing a matrix metalloproteinase domain and a disintegrin domain [Nature, 377, 652-656 (1995); Nature Genet., 5, 151-157 (1993); Nature, 356, 248-252 (1992)]. Examples of known ADAM family proteins are fertilin, epidermal apical protein, cyritestin, MDC (metalloprotease-like, disintegrin-like and cystein-rich protein), meltrin, MS2, and metargidin [Nature, 377, 652-656 (1995); Nature Genet. 5, 151-157 (1993); Nature, 356, 248-252 (1992); Biochem. J., 286, 671-675 (1992); Dev. Growth. Differ., 36, 49-58 (1994); Int. Immunol., 2, 585-591 (1990); J. Biol. Chem., 271, 4593-4596 (1996)].
It was reported that fertilin is involved in an integrin-mediated sperm-egg binding [Nature, 356, 248-252 (1992)], and meltrin is involved in a myotube formation [Nature, 377, 652-656 (1995)]. MDC expressed mainly in a central nervous system is a candidate as a suppresser against a human breast cancer [Nature Genet. 5, 151-157 (1993)], and MS2 serves as a macrophage antigen [Int. Immunol., 2, 585-591 (1990)]. However, little have been known about the physiological roles of these ADAM family proteins.
The mouse ADAMTS-1 protein contains a matrix metalloproteinase domain and a disintegrin domain, and therefore belongs to the ADAM family. However, the mouse ADAMTS-1 protein is different from other known ADAM family proteins in that it further contains thrombospondin domains.
As mentioned, the ADAM family proteins have various kinds of activities such as the involvement of bone or muscle metabolism, suppression of cancer growth, or fertilization, and the thrombospondin exhibits an action to inhibit vascularization and suppresses cancer. Therefore, it is expected that the mouse ADAMTS-1 protein will exhibit unique physiological functions.
The inventors of the present invention attempted to isolate the corresponding human ADAMTS-1 protein. Accordingly, the present inventors designed and prepared various probes on the basis of the base sequence of the known mouse ADAMTS-1 gene, and carried out plaque hybridizations with a human kidney cDNA library so as to obtain a human ADAMTS-1 gene, but a desired gene was not obtained. Then, the present inventors designed and prepared various primers on the basis of the base sequence of the known mouse ADAMTS-1 gene, and carried out PCRs, using the human kidney cDNA library as templates under ordinary conditions, so as to obtain the desired gene, but did not succeed.
Thereafter, the present inventors carried out a PCR of the human kidney cDNA library, using the same primers, but under conditions milder than those ordinarily used, more particularly, an annealing temperature is set lower than an ordinary temperature, and the inventors successfully obtained a novel human ADAMTS-1 gene. The resulting gene was then, expressed in
E. coli
, and the biological activities of the recombinant human ADAMTS-1 protein were examined. Surprisingly, it was found that the novel human ADAMTS-1 protein can decrease the numbers of leukocytes and platelets, and at the same time, increase the number of erythrocytes. Such activities in influencing hematopoietic functions cannot be expected from the structure of the mouse ADAMTS-1 gene which was used as the basis in designing the primers, or from the functions of domains contained in the human ADAMTS-1 protein. The present invention is based on the above findings.
DISCLOSURE OF INVENTION
The present invention relates to a protein characterized by containing an amino acid sequence of SEQ ID NO: 1:
Met Asp Ile Cys Arg Ile Arg Leu Arg Lys Lys Arg Phe Val Ser Ser
Pro Arg Tyr Val Glu Thr Met Leu Val Ala Asp Gln Ser Met Ala Glu
Phe His Gly Ser Gly Leu Lys His Tyr Leu Leu Thr Leu Phe Ser Val
Ala Ala Arg Leu Tyr Lys His Pro Ser Ile Arg Asn Ser Val Ser Leu
Val Val Val Lys Ile Leu Val Ile His Asp Glu Gln Lys Gly Pro Glu
Val Thr Ser Asn Ala Ala Leu Thr Leu Arg Asn Phe Cys Asn Trp Gln
Lys Gln His Asn Pro Pro Ser Asp Arg Asp Ala Glu His Tyr Asp Thr
Ala Ile Leu Phe Thr Arg Gln Asp Leu Cys Gly Ser Gln Thr Cys Asp
Thr Leu Gly Met Ala Asp Val Gly Thr Val Cys Asp Pro Ser Arg Ser
Cys Ser Val Ile Glu Asp Asp Gly Leu Gln Ala Ala Phe Thr Thr Ala
His Glu Leu Gly His Val Phe Asn Met Pro His Asp Asp Ala Lys Gln
Cys Ala Ser Leu Asn Gly Val Asn Gln Asp Ser His Met Met Ala Ser
Met Leu Ser Asn Leu Asp His Ser Gln Pro Trp Ser Pro Cys Ser Ala
Tyr Met Ile Thr Ser Phe Leu Asp Asn Gly His Gly Glu Cys Leu Met
Asp Lys Pro Gln Asn Pro Ile Gln Leu Pro Gly Asp Leu Pro Gly Thr
Leu Tyr Asp Ala Asn Arg Gln Cys Gln Phe Thr Phe Gly Glu Asp Ser
Lys His Cys Pro Asp Ala Ala Ser Thr Cys Ser Thr Leu Trp Cys Thr
Gly Thr Ser Gly Gly Val Leu Val Cys Gln Thr Lys His Phe Pro Trp
Ala Asp Gly Thr Ser Cys Gly Glu Gly Lys Trp Cys Ile Asn Gly Lys
Cys Val Asn Lys Thr Asp Arg Lys His Phe Asp Thr Pro Phe His Gly
Ser Trp Gly Pro Trp Gly Pro Trp Gly Asp Cys Ser Arg Thr Cys Gly
Gly Gly Val Gln Tyr Thr Met Arg Glu Cys Asp Asn Pro Val Pro Lys
Asn Gly Gly Lys Tyr Cys Glu Gly Lys Arg Val Arg Tyr Arg Ser Cys
Asn Leu Glu Asp Cys Pro Asp Asn Asn Gly Lys Thr Phe Arg Glu Glu
Gln Cys Glu Ala His Asn Glu Phe Ser Lys Ala Ser Phe Gly Ser Gly
Pro Ala Val Glu Trp Ile Pro Lys Tyr Ala Gly Val Ser Pro Lys Asp
Arg Cys Lys Leu Ile Cys Gln Ala Lys Gly Ile Gly Tyr Phe Phe Val
Leu Gln Pro Lys Val Val Asp Gly Thr Pro Cys Ser Pro Asp Ser Thr
Ser Val Cys Val Gln Gly Gln Cys Val Lys Ala Gly Cys Asp Arg Ile
Ile Asp Ser Lys Lys Lys Phe Asp Lys Cys Gly Val Cys Gly Gly Asn
Gly Ser Thr Cys Lys Lys Ile Ser Gly Ser Val Thr Ser Ala Lys Pro
Gly Tyr His Asp Ile Val Thr Ile Pro Thr Gly Ala Thr Asn Ile Glu
Val Lys Gln Arg Asn Gln Arg Gly Ser Arg Asn Asn Gly Ser Phe Leu
Ala Ile Lys Ala Ala Asp Gly Thr Tyr Ile Leu Asn Gly Asp Tyr Thr
Leu Ser Thr Leu Glu Gln Asp Ile Met Tyr Lys Gly Val Val Leu Arg
Tyr Ser Gly Ser Ser Ala Ala Leu Glu Arg Ile Arg Ser Phe Ser Pro
Leu Lys Glu Pro Leu Thr Ile Gln Val Leu Thr Val Gly Asn Ala Leu
Arg Pro Lys Ile Lys Tyr Thr Tyr Phe Val Lys Lys Lys Lys Glu Ser
Phe Asn Ala Ile Pro Thr Phe Ser Ala Trp Val Ile Glu Glu Trp Gly
Glu Cys Ser Lys Ser Cys Glu Leu Gly Trp Gln Arg Arg Leu Val Glu
Cys Arg Asp Ile Asn Gly Gln Pro Ala Ser Glu Cys Ala Lys Glu Val
Lys Pro Ala Ser Thr Arg Pro Cys Ala Asp His Pro Cys Pro Gln Trp
Gln Leu Gly Glu Trp Ser Ser Cys Ser Lys Thr Cys Gly Lys Gly Tyr
Lys Lys Arg Ser Leu Lys Cys Leu Ser His Asp Gly Gly Val Leu Ser
His Glu Ser Cys Asp Pro Leu Lys Lys Pro Lys His Phe Ile Asp Phe
Cys Thr Leu Thr Gln Cys Ser.
Further, the present invention relates to the variation of proteins which are equivalent to the protein containing the amino acid sequence of SEQ ID NO: 1.
Further, the present invention relates to a protein characterized by containing a matrix metalloproteinase domain, a disintegrin domain, and a thrombospondin domain, except for a mouse ADAMTS-1 protein.
Further, the present invention relates to a gene characterized by encoding the above novel proteins.
Further, the presen
Hakozaki Michinori
Hirose Kunitaka
Inoguchi Eiji
Ishida Yukako
Ishioka Keiko
Ewoldt Gerald R.
Kureha Chemical Industry Co. Ltd.
LandOfFree
Human ADAMTS-1 protein and pharmaceutical composition does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Human ADAMTS-1 protein and pharmaceutical composition, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Human ADAMTS-1 protein and pharmaceutical composition will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3035634