Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1995-08-24
1999-03-02
Lau, Kawai
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
4352523, 4353201, 514 12, 530324, 530412, C12P 2102, C07K 14435, C07K 136, A61K 3816
Patent
active
058769714
DESCRIPTION:
BRIEF SUMMARY
The invention relates to proteins, which are thrombin inhibitors from the saliva of protostomia.
PRIOR ART
Thrombin has a key function in the thrombus formation in blood vessels. It catalyzes the cleavage of fibrinogen to fibrin. It leads to the formation of blood clots. Further, it induces the aggregation of blood platelets. The enzyme is involved in the pathogenesis of different diseases, such as, e.g., arterial and venous thrombosis or arteriosclerosis.
Therefore, the use of a thrombin inhibitor for treatment of thromboses is promising. The most important previously known thrombin inhibitors are antithrombin III-heparin and hirudin. Antithrombin III is a protein occurring in plasma with a molecular weight of 58 kDa. Antithrombin III binds first to heparin, which is a polysaccharide. Then, the antithrombin III-heparin complex binds to thrombin and inhibits this thrombin. A very stable complex of thrombin and antithrombin III results and antithrombin III is cleaved from thrombin. In addition to thrombin, antithrombin III also inhibits other serine proteases such as, e.g., factor Xa (Pratt, C. W. and Church, F. C. (1991) "Antithrombin: Structure and Function," Seminars in Hematology 28: 3-9).
The protein hirudin was isolated from the Hirudo medicinalis leech. It has a molecular weight of about 7 kDa and binds to thrombin by ionic interaction. It is specific for thrombin (Johnson, P. H. et al. (1989) "Biochemistry and Genetic engineering of hirudin," Seminars in Thrombosis and Hemostasis 15: 302-315).
DESCRIPTION OF THE INVENTION
In addition to the previously mentioned thrombin inhibitors, which are included in the prior art, other inhibitors are necessary which have another mechanism of action or an increased activity.
The invention provides natural proteins or proteins that can be synthetically produced, which are thrombin inhibitors and can be isolated from the saliva of insects which suck the blood of mammals.
Proteins according to the invention which can be isolated from the Triatoma pallidipennis cone-nosed bug are preferred.
The proteins according to the invention can be of natural origin. The proteins are obtained by the saliva being purified. Also, it is possible to isolate the proteins from the salivary glands or to take the cells synthesizing the protein from the salivary glands and to keep them in the culture. The supernatants produced by this cell culture are harvested and worked up. The cell supernatant is purified and the proteins according to the invention are isolated and enriched. All enrichment stages of the isolation and the purification are part of the invention. Preferred are the enrichment stages of the isolation and purification in which the proteins according to the invention can be used for pharmaceutical purposes. Thus, purifications of 50% of the thrombin inhibitor are achieved relative to the total protein, preferred are 85%, more preferred 95% and most preferred 99% of the thrombin inhibitor relative to the total protein.
The invention comprises not only the proteins which can be isolated from Triatoma pallidipennis, but also proteins which can be synthesized from other types of insects. Thus, in addition to the proteins which can be isolated from Triatoma pallidipennis and which are in the group of the most preferred, other proteins are preferred which are derived from Triatoma infestans, Triatoma dimidiata, Triatoma maculata, Rhodnius prolixus, Panstrongylus megistus and Panstrongylus infestans.
It is likewise possible to produce the proteins according to the invention synthetically. Included in this is the protein synthesis according to J. M. SEWART and J. D. YOUNG, San Francisco, 1969 and J. MEIENHOFER, Hormonal Proteins and Peptides, Vol. 2, p. 46, Academic Press (New York), 1973 and E. SCHODER and K. LUBKE, The Peptides, Vol. 1, Academic Press (New York), 1965. The recombinant proteins, which are produced according to known processes, are also included in the synthetically produced proteins. Depending on the host organism, the proteins according to the invention can
REFERENCES:
Hoffman, A. et al. (Mar. 1991) "Isolation and characterization of a thrombin inhibitor from the tick Ixodes ricinus" Pharmazie 46(3):209-212.
Alagon Alejandro
Cuevas-Aguierre Delia
Donner Peter
Haendler Bernard
Hechler Ulrike
Lau Kawai
Schering Aktiengesellschaft
LandOfFree
Thrombin inhibitor from the saliva of protostomia does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Thrombin inhibitor from the saliva of protostomia, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Thrombin inhibitor from the saliva of protostomia will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-421635