Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1996-03-20
1999-01-05
Jacobson, Dian C.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
514 12, 514822, 530350, 530856, 530402, 536 235, 435440, C07K 14435, C07K 1446, C12N 1512, A61K 3816
Patent
active
058561269
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to a peptide having anti-thrombus activity, a method of producing the peptide, and a pharmaceutical composition containing the peptide. In particular, the present invention relates to a peptide originating from a snake venom, the peptide not causing decrease in platelets or thrombocytes (thrombocytopenia) in vivo.
BACKGROUND ART
It is widely known that platelets closely participate in crisis of so-called thrombosis represented by myocardial infarction and cerebral thrombosis ("Platelets", edited by Yamanaka and Yamazaki, Igaku-Syoin, pp. 158-163 (1991)). Recently, it has been reported that the binding between von Willebrand factor as one of blood proteins and glycoprotein Ib located on platelet surfaces is important for platelets to adhere to intravascular subendotherial tissue, which is considered as an early reaction to cause thrombosis (J. P. Cean et al., J. Lab. Clin. Med., 87, 586-596 (1976)).
It is known that the binding between the two species of the proteins does not occur in an ordinary state, but it occurs only when a high shear stress is exerted in vivo (T. T. Vincent et al., Blood, 65, 823-831 (1985)). The methodology to observe the binding exo-vivo includes a widely spread method which uses certain substances such as ristocetin as an antibiotic (M. A. Howard, B. G. Firkin, Thromb. Haemostatis, 26, 362-369 (1971)) and botrocetin as a protein originating from a snake venom (M. S. Read et al., Proc. Natl. Acad. Sci. U.S.A., 75, 4514-4518 (1978)). Platelet aggregation occurs when these substances are added to a suspension of platelets. These aggregation depends on the binding between von Willebrand factor and glycoprotein Ib (M. A. Howard, B. G. Firkin, M. S. Read et al., supra).
Several compounds have been already reported, which exhibit an inhibiting action on the platelet aggregation mediated by ristocetin or botrocetin. Such known compounds include, for example, aurintricarboxylic acid (M. D. Phillips et al., Blood, 72, 1989-1903 (1988)), and dye substances such as aromatic amidino compounds (J. D. Geratz et al., Thromb. Haemostasis, 39, 411-425 (1978)), as well as partial fragment peptides of von Willebrand factor or glycoprotein Ib (Y. Fujimura et al., J. Biol. Chem., 261, 381-385 (1986); K. Titani et al., Proc. Natl. Acad. Sci. U.S.A., 84, 5610-5614 (1987)).
It has been also reported that peptides having similar platelet aggregation-inhibiting activities are present in snake venoms. An international publication pamphlet of WO9208472 describes such peptides from Crotalus horridus horridus and Cerastes cerastes, each peptide comprising two different strands having a molecular weight of about 25 kilodaltons in which the homology is high at least in amino acid sequences on N-terminal side. A platelet aggregation-inhibiting peptide, which has been reported by Peng et al. (M. Peng et al., Blood, 81, 2321-2328 (1993)) as obtained from Echis carinatus, is also extremely similar to the peptides described above in its in vitro activity, molecular weight, etc. Any of the platelet aggregation-inhibiting peptides originating from snake venoms inhibits platelet aggregation mediated by ristocetin or botrocetin at a low concentration of 2 to 5 .mu.g/ml or less in vitro.
The international publication pamphlet of WO9208472 does not describes the anti-thrombus activity upon administration to animals. Accordingly, as described in Example 1 in this specification, the present inventors purified a peptide having equivalent properties to the peptide described in the pamphlet, considering a purification method for the peptide originating from Crotalus horridus horridus described in the pamphlet, in order to investigate the action upon administration to animals. As a result, the present inventors observed almost complete disappearance of platelets in blood upon administration in a small amount of 100 .mu.g/kg. However, the international publication pamphlet of WO9208472 describes neither suggestion nor solution of such a problem upon administration to animals.
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REFERENCES:
patent: 5336667 (1994-08-01), Kirby
patent: 5342830 (1994-08-01), Scarborough
patent: 5679542 (1997-10-01), Scarborough
T.E. Creighton, "Protein Structure a Practical Approach", 1989, IRL Press at Oxford University Press, pp. 155-159.
Yoshihiro Fujimura et al, "Snake Venom Proteins Modulating the Interaction Between von Willebrand Factor and Platelet Glycoprotein lb", 1996, F.K. Schattauer Verlagsgesellschaft mbH (Stuttgart) 76 (5), pp. 633-639.
W. W. Cleland, "Dithiothreitol, A New Protective Reagent For SH Groups", Nov. 4, 1963, From the Department of Biochemistry, University of Wisconsin, Madison, pp. 480-482.
H. Fraenkel-Conrat et al, "The Molecular Weight of Lysozyme After Reduction and Alkylation of the Disulfide Bonds", Feb. 1951, vol. 73, pp. 625-627.
Thomas A. Bewley et al, "Human Pituitary Growth Hormone. XVI. Reduction With Dithiothreitol in the Absence of Urea", 1968, Biochim. Biophys. Acta, 154, pp. 420-422.
Robert K. Andrews, "Binding of a Novel 50-Kilodalton Alboaggregin From Trimeresurus Albolabris and Related Viper Venom Proteins to the Platelet Membrane Glycoprotein Lb-IX-V Complex. Effect On Platelet Aggregation and Glycoprotein Ib-Mediated Platelet Activation", 1996, Biochemistry, vol. 35, 12629-12639.
Peng, M. et al. Blood 81(9):2321-2328 (1993).
Fukuchi Naoyuki
Ishii Koichi
Kito Morikazu
Kobayashi Tsuyoshi
Nagano Mitsuyo
Ajinomoto Co. Inc.
Jacobson Dian C.
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