AU592486B2 – Pancreatic secretory trypsin inhibitor and variants thereof produced by a recombinant host, process, expression vector and recombinant host therefore and pharmaceutical use thereof
– Google Patents
AU592486B2 – Pancreatic secretory trypsin inhibitor and variants thereof produced by a recombinant host, process, expression vector and recombinant host therefore and pharmaceutical use thereof
– Google Patents
Pancreatic secretory trypsin inhibitor and variants thereof produced by a recombinant host, process, expression vector and recombinant host therefore and pharmaceutical use thereof
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Publication number
AU592486B2
AU592486B2
AU10038/88A
AU1003888A
AU592486B2
AU 592486 B2
AU592486 B2
AU 592486B2
AU 10038/88 A
AU10038/88 A
AU 10038/88A
AU 1003888 A
AU1003888 A
AU 1003888A
AU 592486 B2
AU592486 B2
AU 592486B2
Authority
AU
Australia
Prior art keywords
asp
asn
psti
thr
tyr
Prior art date
1987-01-07
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
AU10038/88A
Other versions
AU1003888A
(en
Inventor
Helmut Blocker
Wolfgang Bruns
John Collins
Ronald Frank
Hans Fritz
Friedhelm Maywald
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Helmholtz Zentrum fuer Infektionsforschung HZI GmbH
Original Assignee
Bayer AG
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
1987-01-07
Filing date
1988-01-05
Publication date
1990-01-11
1988-01-05
Application filed by Bayer AG
filed
Critical
Bayer AG
1988-09-01
Publication of AU1003888A
publication
Critical
patent/AU1003888A/en
1990-01-11
Application granted
granted
Critical
1990-01-11
Publication of AU592486B2
publication
Critical
patent/AU592486B2/en
1998-05-21
Assigned to GESELLSCHAFT FUR BIOTECHNOLOGISCHE FORSCHUNG MBH
reassignment
GESELLSCHAFT FUR BIOTECHNOLOGISCHE FORSCHUNG MBH
Alteration of Name(s) in Register under S187
Assignors: BAYER AKTIENGESELLSCHAFT
2008-01-05
Anticipated expiration
legal-status
Critical
Status
Ceased
legal-status
Critical
Current
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Classifications
C—CHEMISTRY; METALLURGY
C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
C—CHEMISTRY; METALLURGY
C07—ORGANIC CHEMISTRY
C07K—PEPTIDES
C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
C07K14/81—Protease inhibitors
C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
C07K14/811—Serine protease (E.C. 3.4.21) inhibitors
C07K14/8135—Kazal type inhibitors, e.g. pancreatic secretory inhibitor, ovomucoid
A—HUMAN NECESSITIES
A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
A—HUMAN NECESSITIES
A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
A61P37/00—Drugs for immunological or allergic disorders
A61P37/08—Antiallergic agents
A—HUMAN NECESSITIES
A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
A61K38/00—Medicinal preparations containing peptides
C—CHEMISTRY; METALLURGY
C07—ORGANIC CHEMISTRY
C07K—PEPTIDES
C07K2319/00—Fusion polypeptide
C—CHEMISTRY; METALLURGY
C07—ORGANIC CHEMISTRY
C07K—PEPTIDES
C07K2319/00—Fusion polypeptide
C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
C—CHEMISTRY; METALLURGY
C07—ORGANIC CHEMISTRY
C07K—PEPTIDES
C07K2319/00—Fusion polypeptide
C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
C07K2319/036—Fusion polypeptide containing a localisation/targetting motif targeting to the medium outside of the cell, e.g. type III secretion
Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
Y10S930/00—Peptide or protein sequence
Y10S930/01—Peptide or protein sequence
Y10S930/26—Containing cys-cys disulfide bridge between nonadjacent cysteine residues
Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
Y10S930/00—Peptide or protein sequence
Y10S930/01—Peptide or protein sequence
Y10S930/27—Cyclic peptide or cyclic protein
Abstract
A peptide which essentially has the amino acid sequence of pancreatic secretory trypsin inhibitor (PSTI) and variants of this peptide in which one or more of the amino acids of the original sequence are replaced by other amino acids are described. These peptides are modified in an advantageous manner with respect to the specificity of their inhibitor action. A process for the preparation of the peptides and their pharmaceutical use is additionally described.
Description
‘1 592406 Form PATENTS ACT 1952-1 973 COMPLETE SPECIFICATION
(ORIGINAL)
FOR OFFICE USE Class: Int. Cl: Application Number: Lodged: Complete Specification-Lodged: Accepted: Published: Priority: Titii dc~rn»Cfl’ conltainsJ b nsflt nts mde uAgd& S( 4Zofl 49.
sdIs WtV»c Wr r Relaied Art; TO BE COMPLETED BY APPLICANT Name of Applicant: k, 0v~ 6_0~ C-6_41_ C J: -f Address of Applicant: D S 09 Levv-K’ser I 6- erw-n c4n/ A ctual Inventor Zc~i’c Co k r S H e m i rz o ci e c, R o n’c»J Pf-c nr’K Address for Service: e& S $reeV 1 eye> K~eW Sout-H J ,(e Complete Specification for thr@ Invention entitled:- eAtc~QS.jLerC. is rc,.a7OE TLYPSi1J ‘WNUakvoR FiMb V~,4MtAArS -TevteO; Pf00UrCGb rs»j A t -_*HSA I PINT kO. 7r \)cVc–rorL AND1J Peo iM A#T Ros.7 7hE46eefo~ m J hr ~ingstatement Is a full description of this Invention, Including the best method of performing It knoWn to me:i- N -The description Is to be typed In double spacing, pica type face, In an area not eWeeding 2~0 mm In depth and 160 mm In width, !8 IVon tough white paper of good quality end It Is to be Inserted Inside this form, r. 0, Atkinsonl, Government Printer, Canberra r 1i ii u la panlcres.. se4-caory YPss> >nh\iy-or Enzyme inhibit and variants thereof produced by a recombinant host, process, expression vector and recombinant host therefore and pharmaceutical use thereof The present invention relates to a microbially produced peptide having essentially the amino acid sequence of human pancreactic secretory trypsin inhibitor (h-PSTI). The present invention further relates to variants of such peptide wherein one or more of the amino acids in the original sequence are replaced by other amino acids. These peptides show an S’ advantageously modified specificity in their inhibitory action. A method of preparation of the peptides and their pharmaceutical use is also described.
2 Introduction tr, i t The lysosomal elastase (leukocyte elastase); J.G.
Bieth (1986) p. 217 320 (in Regulation of Matrix Accumulation, Mecham ed., Academic Press, Orlando) from l polymorphonuclear granulocytes is a potent intracellular protease which is stored in lysosomes and fulfills its physiological function, the intracellular protein breakdown, in phagolysosomes. The major functional role of lysosomal proteases (elastase, cathepsin G, etc.; H.
Fritz et al (1984) (in Selected Topics in Clinical Enzymology, Goldberg and Werner ed., Walter de Gruyter, Le A 24 828 Foreign Countries r, 1 r I «ol~~ 2 Berlin vol. 2, p. 305 328) is the degradation of phagocytized material from either the organism itself metabolic products, injured tissue) or from invasive organisms (bacteria, viruses, molds, etc.).
Upon release into the extracellular space (blood or interstitial fluid) elastase is rapidly bound by potent endogenous inhibitors such as al-PI (al-protease inhibitor; J. Travis and G.S. Salvesen, (1983), Ann. Rev. Biochem p. 655 709,) in plasma, and/or antileukoprotease (also called HUSI-I, human seminal t r proteinase inhibitor); H. Schiessler et al (1978) p.
195-207, in Neutral Proteases of Human Polymorphonuclear *Leukocytes, Havemann and Janoff ed., Urban Schwarzenberg, Baltimore) in mucous secretions.
Due to a hereditary oc(-PI-deficiency Bieth, 1986) or as a consequence of massive extracellular release of elastase (in acute and chronic inflammations, polytrauma or shock; H. Fritz et al, 1984) the protection of the organism against the degradative potential of elastase by natural protease inhibitors is insufficient. An excessive and locally even total consumption of the endogenous protease inhibitors is caused by complex formation with elastase, (ii) proteolytic inactivation by various lysosomal proteases, and (iii) particularly by oxidative inactivation (I-PI) Bieth (1986); H. Fritz et al. (1984); J. Travis and G,S. Salvesen (1983); see above).
The consequence is an extensive proteolytic degradation of connective tissues, as well as of humoral proteins including coagulation-, fibrinolysis-, and com Le A 24 828 3 plement factors by elastase and other lysosomal proteases cathepsin G) leading to severe clinical symptoms like emphysema, shock lung, adult respiratory distress syndrome, coagulation disorders, kidney and liver failure, etc. (additionally to the above references: Neue Wege in der EntzUndungsdiagnostik, PMN Elastase; M. Jochum et al (1985), GIT Verlag, Darmstadt; C.T. Lee etal, (1981), N. Engl. J. Med., 304, 192-196; W.W. McGuire eL al, (1982), J. Clin. Invest.
69, 543).
Elastase contributes also to the local inflammatory event going on in rheumatoid arthritis, the degradation of connective tissue constituents (K.
Kleesiek et al (1985) in Neue Wege in der Entzundungsdiagnostik, PMN Elastase, p. 71 82).
The extracel]ular release of elastase after severe injuries or in diseases like septic shock, shock lung etc. can be monitored routinely by means of enzyme immnoassays Neumann and M. Jochum, (1984), p. 184 -195 in Methods of Enzymatic Analysis, Bergmeyer (ed), Verlag Chemie, Weinheim).
In experimental models of sepsis and emphysema, synthetic elastase inhibitors Powers, Am. Rev.
Respir. Dis., (1983), 127, 554 558) and natural inhibitors from animals such as eglin C Schnebli et al, (1985), Eur. J. Respir. Dis. 66, Suppl. 139 p.
66 70) have proven to be therapeutically useful. The application of a protease inhibitor of human origin would be preferable in order to avoid toxic side effects and especially allergic reactions when a prolonged Lo A 24 828 r 4 therapy is indicated, in the treatment of (a-PI S deficiency (emphysema). Since the human al-PI is a glycoprotein of high molecular weight, its production in sufficient amounts by means of gene technology is not feasible in the near future.
The antileucoprotease or HUSI-I Schiessler et al, 1978; and U. SeemUller et al (1986) FEBS Letters 199, 43 48) has a MW of 14 000 Dalton and consists of j two active domains, one directed against elastase, the Sother against trypsin. Hence this type of inhibitor has a relative low selectivity and is not a specific elastase inhibitor. Inhibition of trypsin or trypsin-like enzymes is normally not intended in the indications given above.
The human pancreas secretes the PSTI, a protease inhibitor of low MW (6.2 kd) which specifically inhibits trypsin, i.e. it possesses a high selectivity for one specific type of protease.
One advantage presented in this invention consists in substitution by recombinant DNA technology of only one (or a few) amino acids in the PSTI yielding PSTI variants which were shown to be highly effective protease inhibitors with high specificity for leukocyte elastase. Moreover, due to its relatively low MW the elastase PSTI derivative complex should pass through the kidney as well, Therefore, elimination of extracellularly released elastase will be highly efficient. The fact that PSTI is of human origin, taken together with its low MW lead applicants to expect that the clinical application of PSTI derivatives will be devoid of complications due Le A 24 828 i i
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to recognition of these substances as foreign proteins by the immune system. Another essential advantage of PSTI compared to
