WO1998056810A2 - Lus-1 human protein, its production and use - Google Patents
Lus-1 human protein, its production and use Download PDFInfo
- Publication number
- WO1998056810A2 WO1998056810A2 PCT/EP1998/003460 EP9803460W WO9856810A2 WO 1998056810 A2 WO1998056810 A2 WO 1998056810A2 EP 9803460 W EP9803460 W EP 9803460W WO 9856810 A2 WO9856810 A2 WO 9856810A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- cys
- thr
- ser
- ala
- leu
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4702—Regulators; Modulating activity
- C07K14/4703—Inhibitors; Suppressors
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/05—Animals comprising random inserted nucleic acids (transgenic)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Definitions
- This disulfide pattern was obtained by proteolytic cleavage with various endoproteases and sequencing of the fragments obtained.
- the corresponding fragments were sequenced by a combination of electrospray mass spectroscopy and Edman protein sequencing. Both human hemofiltrate and urine-derived LUS-I had identical chromatographic properties, molecular weights, and disulfide patterns.
- a further embodiment of the invention is represented by pharmaceutical compositions which contain the peptides according to the invention in a pharmaceutical dosage form.
- the preferred amount of the peptides according to the invention to be administered is between 1 ⁇ g and 1 g per dose unit when 1 to 5 doses of the drug are administered per day.
- nucleic acids which code for the peptides according to the invention can derive nucleic acids which code for the peptides according to the invention from the sequence of the peptides according to the invention using the genetic code. For expression in recombinant organisms, those codons are chosen which correspond to the codon usage of the transformed organism.
- Antisense nucleotides i.e. Nucleotides which bind to the nucleic acid sequences according to the invention under stringent conditions have, in particular, those sequences which can bind to parts of the mRNA in the target system.
- the person skilled in the art can determine the sequence of the mRNA from the peptide sequence using known methods.
- Antibodies that bind to the peptides according to the invention can be obtained in a simple manner by immunizing animals become.
- the peptides are optionally injected into the animals together with other auxiliary substances at intervals of several weeks, and then antibodies are isolated from the blood.
- Preferred animals for this are rabbits and mice.
- Known methods can be used to obtain immortal cells from the antibody-producing mouse cells, which allow the production of monoclonal antibodies.
- the transgenic mammals according to the invention with gene efficiency LUS-1 are obtained by methods known to the person skilled in the art. In general, the homologous recombination between DNA sequences is used. The vectors required for this can be constructed in a simple manner if the gene sequence is known. If the mRNA sequence is known, the gene can be found, for example, likewise by means of a polymerase chain reaction, and the amplified gene fragment can be elucidated by means of sequencing.
- the proteins, nucleic acids and antisense nucleotides, antibodies and inhibitors according to the invention are suitable for the production of diagnostic agents.
- the antibodies in particular enable the expression of the protein according to the invention to be determined in tissue or body fluids, for example by means of an immunoassay such as the known ELISA.
- an immunoassay such as the known ELISA.
- the nucleic acids according to the invention it can be analyzed - by means of amplification reactions such as polymerase chain reactions - whether there are gene diseases which can then be treated with the help of the medicaments according to the invention.
Abstract
This invention relates to protein having the following amino acid sequence, in accordance with ID Sequence ID N° 1: NH2-Leu-Lys-Cys-Tyr-Thr- Cys-Lys-Glu-Pro-Met- Thr-Ser-Ala-Ser-Cys-Arg- Thr-Ile-Thr-Arg- Cys-Lys-Pro-Glu-Asp-Thr- Ala-Cys-Met-Thr-Thr-Leu- Val-Thr-Val-Glu- Ala-Glu-Tyr-Pro-Phe- Asn-Gln-Ser-Pro- Val-Val-Thr-Arg- Ser-Cys-Ser-Ser- Ser-Cys-Val-Ala-Thr- Asp-Pro-Asp-Ser- Ile-Gly-Ala-Ala- His-Leu-Ile-Phe- Cys-Cys-Phe-Arg- Asp-Leu-Cys-Asn- Ser-Glu-Leu-COOH (LUS-I). It also relates to its cyclic, glycosylated, phosphorylated, acetylated, amidated derivates, and/or its derivatives containing cross-linkages of side chains, as well as fragments having the biological activity of LUS-I.
Description
Humanes Protein LUS-I, seine Herstellung und Verwendung Human protein LUS-I, its production and use
Die vorliegende Erfindung betrifft Peptide und Peptidderivate mit biologischer Aktivität.The present invention relates to peptides and peptide derivatives with biological activity.
Ridge und Sloane publizierten ein anti-neoplastisches Urinpeptid (R. J. Ridge & N.H. Sloane, Cytokine 8 (1996) , 1-5) mit einer dort angegebenen Struktur und einem Molekulargewicht in der Größenordnung von 16 kDa . Dieses Protein ist auch Gegenstand der US-PS 5,298,604.Ridge and Sloane published an anti-neoplastic urine peptide (R. J. Ridge & N.H. Sloane, Cytokine 8 (1996), 1-5) with a structure given there and a molecular weight in the order of 16 kDa. This protein is also the subject of US Pat. No. 5,298,604.
Erfindungsgemäß wurde ein Protein mit der Aminosäuresequenz der FormelAccording to the invention, a protein with the amino acid sequence of the formula
H2-Leu-Lys-Cys-Tyr-Thr-Cys-Lys-Glu-Pro-Met-Thr-Ser-Ala-Ser-Cys- Arg-Thr-Ile-Thr-Arg-Cys-Lys-Pro-Glu-Asp-Thr-Ala-Cys-Met-Thr- Thr-Leu-Val-Thr-Val-Glu-Ala-Glu-Tyr-Pro-Phe-Asn-Gln-Ser-Pro- Val -Val-Thr-Arg-Ser-Cys-Ser-Ser-Ser-Cys-Val-Ala-Thr-Asp-Pro- Asp-Ser-Ile-Gly-Ala-Ala-His-Leu-Ile-Phe-Cys-Cys-Phe-Arg-Asp- Leu-Cys-Asn-Ser-Glu-Leu-COOH (LUS-I) gefunden.H 2 -Leu-Lys-Cys-Tyr-Thr-Cys-Lys-Glu-Pro-Met-Thr-Ser-Ala-Ser-Cys-Arg-Thr-Ile-Thr-Arg-Cys-Lys-Pro-Glu -Asp-Thr-Ala-Cys-Met-Thr-Thr-Leu-Val-Thr-Val-Glu-Ala-Glu-Tyr-Pro-Phe-Asn-Gln-Ser-Pro-Val -Val-Thr-Arg -Ser-Cys-Ser-Ser-Ser-Cys-Val-Ala-Thr-Asp-Pro Asp-Ser-Ile-Gly-Ala-Ala-His-Leu-Ile-Phe-Cys-Cys-Phe-Arg -Asp- Leu-Cys-Asn-Ser-Glu-Leu-COOH (LUS-I) found.
Auch die cyclischen, glykosylierten, phosphorylierten, acetylierten, amidierten und/oder Verknüpfungen von Seitenketten enthaltenden Derivate sowie Fragmente mit der biologischen Aktivität von LUS-I sind Gegenstand der Erfindung.The invention also relates to the derivatives containing cyclic, glycosylated, phosphorylated, acetylated, amidated and / or linkages of side chains and fragments with the biological activity of LUS-I.
Entsprechende Derivate sind durch dem Fachmann an sich bekannte Verfahren, insbesondere durch Synthese der Peptide an festen Phasen nach Merryfield erhältlich. Alternativ hierzu kann auch das erhaltene Peptid chemisch modifiziert werden. Entsprechende Derivate können stabiler gegenüber einer Proteolyse sein und/ oder veränderter Halbwertszeiten in einem Organismus aufweisen.
In einer bevorzugten Ausführungsform bildet das Cystein in Sequenzposition 3 mit dem Cystein in Position 28, das Cystein in Position 6 mit dem Cystein in Position 15, das Cystein in Position 21 mit dem Cystein in Position 51 oder 55, das Cystein in Position 77 mit dem Cystein in Position 71 oder 72 und/oder das Cystein in Position 51 oder 55 mit dem Cystein in Position 71 oder 72 eine intramolekulare Disulfidbrücke. Dieses Disulfid- muster wurde mittels proteolytischer Spaltung mit verschiedenen Endoproteasen und Sequenzierung der erhaltenen Bruchstücke erhalten. Die entsprechenden Fragmente wurden durch Kombination von Elektrospray-Massenspektroskopie und Proteinsequenzierung nach Edman sequenziert. Sowohl aus menschlichem Hämofiltrat als auch aus Urin gewonnenes LUS-I wiesen identische chromatographische Eigenschaften, Molekulargewichte und Disulfidmuster auf.Corresponding derivatives can be obtained by methods known per se to the person skilled in the art, in particular by synthesis of the peptides on solid phases according to Merryfield. Alternatively, the peptide obtained can also be chemically modified. Corresponding derivatives can be more stable to proteolysis and / or have changed half-lives in an organism. In a preferred embodiment, the cysteine in sequence position 3 forms with the cysteine in position 28, the cysteine in position 6 with the cysteine in position 15, the cysteine in position 21 with the cysteine in position 51 or 55, the cysteine in position 77 with the Cysteine in position 71 or 72 and / or the cysteine in position 51 or 55 with the cysteine in position 71 or 72 an intramolecular disulfide bridge. This disulfide pattern was obtained by proteolytic cleavage with various endoproteases and sequencing of the fragments obtained. The corresponding fragments were sequenced by a combination of electrospray mass spectroscopy and Edman protein sequencing. Both human hemofiltrate and urine-derived LUS-I had identical chromatographic properties, molecular weights, and disulfide patterns.
Das erfindungsgemäße Protein läßt sich aus humanem Blutfiltrat oder Urin mittels chromatographischer Methoden in reiner Form isolieren oder durch Festphasenpeptidsynthese oder rekombinante Expression in Mikroorganismen herstellen.The protein according to the invention can be isolated in pure form from human blood filtrate or urine by means of chromatographic methods or can be produced by solid phase peptide synthesis or recombinant expression in microorganisms.
Die Bestimmung des relativen Molekulargewichts durch Elektro- spray-Massenspektrometrie von isoliertem LUS-I aus menschlichem Blutfiltrat ergab 8845 Da und stimmt mit dem berechneten Molekulargewicht der bestimmten Aminosäuresequenz von 8843 Da überein. Die bevorzugte Struktur des Peptides LUS-I besteht darin, da/3 alle in seiner Aminosäuresequenz enthaltenen Cysteinreste Bestandteile von Disulfidbrücken sind.The determination of the relative molecular weight by electrospray mass spectrometry of isolated LUS-I from human blood filtrate gave 8845 Da and corresponds to the calculated molecular weight of the determined amino acid sequence of 8843 Da. The preferred structure of the peptide LUS-I is that all the cysteine residues contained in its amino acid sequence are components of disulfide bridges.
Disulfidbrücken bilden sich im Rahmen der Faltung des Peptides im allgemeinen eigenständig. Die Faltung eines durch chemische Synthese oder rekombinante Expression erhaltenen Peptides kann durch geeignete Agenzien unterstützt werden. Entsprechende Techniken sind dem Fachmann für die Aufarbeitung von sogenannten "inclusion bodies" aus Bakterien bekannt.Disulfide bridges generally form independently during the folding of the peptide. The folding of a peptide obtained by chemical synthesis or recombinant expression can be supported by suitable agents. Appropriate techniques are known to the person skilled in the art for processing so-called "inclusion bodies" from bacteria.
Das erfindungsgemäße Protein LUS-I wird in einer nicht membrangebundenen Form aus seinen Ursprungsgeweben in das Blut sezer-
niert. Wie molekularbiologische Befunde ergeben, wird das erfindungsgemäße Protein insbesondere in Tonsillen, der Mundschleimhaut, Augenschleimhäuten, Vaginalepithel, Urethra, Magenschleimhaut, Nebenniere und Niere exprimiert .The protein LUS-I according to the invention is secreted in a non-membrane-bound form from its original tissues into the blood. kidney. As molecular-biological findings show, the protein according to the invention is expressed in particular in tonsils, the oral mucosa, mucous membranes of the eyes, vaginal epithelium, urethra, gastric mucosa, adrenal gland and kidney.
Das erfindungsgemäße Protein eignet sich insbesondere zur Herstellung eines Arzneimittels zur Behandlung von bakteriellen und viralen Effekten, der Über- oder Unterexpression von LUS-I, von Krebserkrankungen, wie Krebserkrankungen des Gebärmutterhalses, das kleinzellige Bronchialkarzinom, Pankreaskarzinom, Mammakar- zinom und Melanome sowie von Autoimmunerkrankungen, angioneuro- tischem Ödem, Asthma bronchiale und paroxysmaler nächtlicher Hämoglobinurie .The protein according to the invention is particularly suitable for the production of a medicament for the treatment of bacterial and viral effects, the over- or under-expression of LUS-I, cancer diseases, such as cancer of the cervix, small cell bronchial carcinoma, pancreatic carcinoma, breast cancer and melanoma, and autoimmune diseases , angioneurotic edema, bronchial asthma and paroxysmal nocturnal hemoglobinuria.
Eine weitere Ausführungsform der Erfindung stellen Arzneimittel dar, die die erfindungsgemäßen Peptide in einer pharmazeutisch üblichen Darreichungsform enthalten.A further embodiment of the invention is represented by pharmaceutical compositions which contain the peptides according to the invention in a pharmaceutical dosage form.
Mögliche pharmazeutische Darreichungsformen sind insbesondere solche, die dem Fachmann bekannt sind für die intravenöse, intraarterielle, intramuskuläre, orale, nasale oder transpul - monare Applikation. Dabei sind insbesondere solche Darreichungsformen bevorzugt, die einen Abbau der erfindungsgemäßen Peptide durch Proteolyse nach der Verabreichung verhindern oder verzögern. Für eine orale Applikation wird es daher bevorzugt, eine Darreichungsform zu wählen, die die Einwirkung von Verdauungsenzymen auf die erfindungsgemäße Peptide vermeidet. Die zur Herstellung dieser Darreichungsformen zu verwendenen Hilfsstoffe sind dem Fachmann bekannt. Es sind insbesondere solche Hilfsstoffe bevorzugt, die keine toxischen Wirkungen aufweisen.Possible pharmaceutical dosage forms are in particular those which are known to the person skilled in the art for intravenous, intraarterial, intramuscular, oral, nasal or transpulmonary administration. Dosage forms which prevent or delay degradation of the peptides according to the invention by proteolysis after administration are particularly preferred. For oral administration, it is therefore preferred to choose a dosage form which avoids the action of digestive enzymes on the peptides according to the invention. The auxiliaries to be used to prepare these dosage forms are known to the person skilled in the art. Auxiliaries which have no toxic effects are particularly preferred.
Die bevorzugte, zu verabreichende Menge der erfindungsgemäßen Peptide liegt zwischen 1 μg und 1 g pro Dosiseinheit bei einer Verabreichung von 1 bis 5 Dosen des Arzneimittels pro Tag.The preferred amount of the peptides according to the invention to be administered is between 1 μg and 1 g per dose unit when 1 to 5 doses of the drug are administered per day.
Weitere bevorzugte Ausführungsformen der Erfindung sind Nukleinsäuren, die für die erfindungsgemäßen Peptide kodieren, Antisen-
senukleotide, die unter stringenten Bedingungen an Nukleinsäu- resequenzen binden, die für die erfindungsgemäßen Peptide kodieren, Antikörper, die an die erfindungsgemäßen Peptide binden, Inhibitoren, die die biologische Aktivität der erfindungsgemäßen Peptide hemmen und Inhibitoren, die die Expression von LUS-I hemmen sowie transgene Säugetiere mit Gendefizienz für LUS-I, die sich insbesondere zur Tumorforschung eignen, zur Untersuchung des Einflusses von LUS-I auf die Bildung und Entwicklung von Tumoren.Further preferred embodiments of the invention are nucleic acids which code for the peptides according to the invention, senucleotides which bind under stringent conditions to nucleic acid sequences which code for the peptides according to the invention, antibodies which bind to the peptides according to the invention, inhibitors which inhibit the biological activity of the peptides according to the invention and inhibitors which inhibit the expression of LUS-I and Transgenic mammals with gene deficiency for LUS-I, which are particularly suitable for tumor research, to investigate the influence of LUS-I on the formation and development of tumors.
Nukleinsäuren, die für die erfindungsgemäßen Peptide kodieren, kann der Fachmann unter Anwendung des genetischen Codes aus der Sequenz der erfindungsgemäßen Peptide ableiten. Für eine Expression in rekombinanten Organismen werden insbesondere solche Codons gewählt, die dem Codon Usage des transformierten Organismus entsprechen. Antisensenucleotide, d.h. Nucleotide, die unter stringenten Bedingungen an die erfindungsgemäßen Nukleinsäurese- quenzen binden, weisen insbesondere solche Sequenzen auf, die mit Teilen der mRNA im ZielSystem eine Bindung eingehen können. Aus der Peptidsequenz kann der Fachmann mit bekannten Methoden die Sequenz der mRNA ermitteln. Beispielsweise können hierzu aus geringvarianten Bereichen Primergemische abgeleitet werden, mit denen mit Hilfe der Polymerasekettenreaktion, die für die erfindungsgemäßen Peptide kodierende mRNA (nach Umschreibung in cDNA mittels reverser Transkriptase) , amplifiziert werden können. Mit bekannten Methoden der DNA-Sequenzierung kann so die Sequenz der mRNA bestimmt werden.The person skilled in the art can derive nucleic acids which code for the peptides according to the invention from the sequence of the peptides according to the invention using the genetic code. For expression in recombinant organisms, those codons are chosen which correspond to the codon usage of the transformed organism. Antisense nucleotides, i.e. Nucleotides which bind to the nucleic acid sequences according to the invention under stringent conditions have, in particular, those sequences which can bind to parts of the mRNA in the target system. The person skilled in the art can determine the sequence of the mRNA from the peptide sequence using known methods. For this purpose, for example, primer mixtures can be derived from small variants, with which the mRNA coding for the peptides according to the invention (after rewriting into cDNA by means of reverse transcriptase) can be amplified with the aid of the polymerase chain reaction. The sequence of the mRNA can thus be determined using known methods of DNA sequencing.
In bevorzugten Ausführungsformen weisen die Antisensenukleotide Modifikationen auf, die den Abbau der Nukleotide durch endogene Substanzen, insbesondere Nukleasen verzögern. Solche Modifikationen sind dem Fachmann bekannt, insbesondere durch Modifikation der Phosphatbindungen wie beispielsweise Thiophosphate, Methylphosphate, H-Phosphonate, Phosphotriester usw.In preferred embodiments, the antisense nucleotides have modifications which delay the degradation of the nucleotides by endogenous substances, in particular nucleases. Such modifications are known to the person skilled in the art, in particular by modification of the phosphate bonds such as, for example, thiophosphates, methylphosphates, H-phosphonates, phosphotriester, etc.
Antikörper, die an die erfindungsgemäßen Peptide binden, können in einfacher Weise durch Immunisierung von Tieren erhalten
werden. Dazu werden die Peptide gegebenenfalls zusammen mit weiteren Hilfsstoffen in mehrwöchigem Abstand den Tieren injiziert und anschließend Antikörpern aus dem Blut isoliert. Bevorzugte Tiere sind hierfür Kaninchen und Mäuse. Durch bekannte Verfahren können aus den antikörperproduzierenden Mäusezellen immortale Zellen erhalten werden, die die Produktion von mono- klonalen Antikörper erlauben.Antibodies that bind to the peptides according to the invention can be obtained in a simple manner by immunizing animals become. For this purpose, the peptides are optionally injected into the animals together with other auxiliary substances at intervals of several weeks, and then antibodies are isolated from the blood. Preferred animals for this are rabbits and mice. Known methods can be used to obtain immortal cells from the antibody-producing mouse cells, which allow the production of monoclonal antibodies.
Inhibitoren, die die biologische Aktivität der Peptide hemmen, sind insbesondere solche, die nicht-kovalent oder kovalent an das Enzym binden. Dies können Antikörper sein oder auch niedermolekulare Moleküle. Inhibitoren, die die Expression von LUS-1 hemmen, sind Substanzen, die die Transkription oder Translation des Genes hemmen. Dies können beispielsweise Antisensennuk- leotide sein.Inhibitors that inhibit the biological activity of the peptides are, in particular, those that bind non-covalently or covalently to the enzyme. These can be antibodies or also small molecules. Inhibitors that inhibit the expression of LUS-1 are substances that inhibit the transcription or translation of the gene. These can be antisense nucleotides, for example.
Die erfindungsgemäßen transgenen Säugetiere mit Gendeffizienz LUS-1 werden nach dem Fachmann bekannten Methoden erhalten. Dabei wird im allgemeinen die homologe Rekombination zwischen DNA-Sequenzen ausgenutzt. Die dazu notwendigen Vektoren lassen sich bei Kenntnis der Gensequenz in einfacher Weise konstruieren. Bei Kenntnis der mRNA-Sequenz kann beispielsweise - ebenfalls durch Polymerasekettenreaktion - das Gen aufgefunden werden und das amplifizierte Genfragment mittels Sequenzierung aufgeklärt werden.The transgenic mammals according to the invention with gene efficiency LUS-1 are obtained by methods known to the person skilled in the art. In general, the homologous recombination between DNA sequences is used. The vectors required for this can be constructed in a simple manner if the gene sequence is known. If the mRNA sequence is known, the gene can be found, for example, likewise by means of a polymerase chain reaction, and the amplified gene fragment can be elucidated by means of sequencing.
Die erfindungsgemäßen Nukleinsäuren eigenen sich auch zur Herstellung eines Medikamentes zur Behandlung von somatischen und nicht somatischen Generkrankungen, indem die fehlende oder nicht korrekte Expression des LUS-1 korrigiert wird.The nucleic acids according to the invention are also suitable for the manufacture of a medicament for the treatment of somatic and non-somatic genetic diseases by correcting the missing or incorrect expression of the LUS-1.
Desweiteren eignen sich die erfindungsgemäßen Proteine, Nukleinsäuren und Antisensenukleotide, Antikörper und Inhibitoren zur Herstellung von Diagnostikmitteln. Insbesondere die Antikörper ermöglichen dabei eine Bestimmung der Expression des erfindungsgemäßen Proteins in Gewebe- oder Körperflüssigkeiten, beispielsweise mittels eines Immunoassays wie des bekannten ELISA.
Mit Hilfe der erfindungsgemäßen Nukleinsäuren kann - durch Amplifikationsreaktionen wie Polymerasekettenreaktione - analysiert werden, ob Generkrankungen vorliegen, die dann mit Hilfe der erfindungsgemäßen Arzneimittel behandelt werden können.
Furthermore, the proteins, nucleic acids and antisense nucleotides, antibodies and inhibitors according to the invention are suitable for the production of diagnostic agents. The antibodies in particular enable the expression of the protein according to the invention to be determined in tissue or body fluids, for example by means of an immunoassay such as the known ELISA. With the aid of the nucleic acids according to the invention, it can be analyzed - by means of amplification reactions such as polymerase chain reactions - whether there are gene diseases which can then be treated with the help of the medicaments according to the invention.
SEQUENZPROTOKOLLSEQUENCE LOG
(1) ALLGEMEINE ANGABEN:(1. GENERAL INFORMATION:
(i) ANMELDER:(i) APPLICANT:
(A) NAME: Wolf -Georg Forssmann(A) NAME: Wolf -Georg Forssmann
(B) STRASSE: Feodor-Lynen-Strasse 31(B) STREET: Feodor-Lynen-Strasse 31
(C) ORT: Hannover(C) LOCATION: Hanover
(E) LAND: Deutschland(E) COUNTRY: Germany
(F) POSTLEITZAHL: 30625(F) POSTAL NUMBER: 30625
(ii) BEZEICHNUNG DER ERFINDUNG: Humanes Protein LUS-I, seine Herstellung und Verwendung(ii) DESCRIPTION OF THE INVENTION: Human protein LUS-I, its production and use
(iii) ANZAHL DER SEQUENZEN: 1(iii) NUMBER OF SEQUENCES: 1
(iv) COMPUTER-LESBARE FASSUNG:(iv) COMPUTER READABLE VERSION:
(A) DATENTRÄGER: Floppy disk(A) DISK: Floppy disk
(B) COMPUTER: IBM PC compatible(B) COMPUTER: IBM PC compatible
(C) BETRIEBSSYSTEM: PC-DOS/MS-DOS(C) OPERATING SYSTEM: PC-DOS / MS-DOS
(D) SOFTWARE: Patentin Release #1.0, Version #1.30 (EPA)(D) SOFTWARE: Patentin Release # 1.0, Version # 1.30 (EPA)
(2) ANGABEN ZU SEQ ID NO: 1:(2) INFORMATION ON SEQ ID NO: 1:
(i) SEQUENZKENNZEICHEN:(i) SEQUENCE LABEL:
(A) LÄNGE: 81 Aminosäuren(A) LENGTH: 81 amino acids
(B) ART: Aminosäure(B) TYPE: amino acid
(C) STRANGFORM: nicht bekannt(C) STRANDFORM: not known
(D) TOPOLOGIE: nicht bekannt(D) TOPOLOGY: not known
(ii) ART DES MOLEKÜLS: Peptid(ii) MOLECULE TYPE: Peptide
(xi) SEQUENZBESCHREIBUNG: SEQ ID NO: 1:(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1:
Leu Lys Cys Tyr Thr Cys Lys Glu Pro Met Thr Ser Ala Ser Cys ArgLeu Lys Cys Tyr Thr Cys Lys Glu Pro Met Thr Ser Ala Ser Cys Arg
1 5 10 151 5 10 15
Thr Ile Thr Arg Cys Lys Pro Glu Asp Thr Ala Cys Met Thr Thr LeuThr Ile Thr Arg Cys Lys Pro Glu Asp Thr Ala Cys Met Thr Thr Leu
20 25 3020 25 30
Val Thr Val Glu Ala Glu Tyr Pro Phe Asn Gin Ser Pro Val Val Thr 35 40 45
Arg Ser Cys Ser Ser Ser Cys Val Ala Thr Asp Pro Asp Ser Ile GlyVal Thr Val Glu Ala Glu Tyr Pro Phe Asn Gin Ser Pro Val Val Thr 35 40 45 Arg Ser Cys Ser Ser Ser Cys Val Ala Thr Asp Pro Asp Ser Ile Gly
50 55 6050 55 60
Ala Ala His Leu Ile Phe Cys Cys Phe Arg Asp Leu Cys Asn Ser Glu 65 70 75 80Ala Ala His Leu Ile Phe Cys Cys Phe Arg Asp Leu Cys Asn Ser Glu 65 70 75 80
Leu
Leu
Claims
1. Protein mit der Aminosäuresequenz gemäß der Sequenz ID No. 11. Protein with the amino acid sequence according to the sequence ID No. 1
NH2-Leu-Lys-Cys-Tyr-Thr-Cys-Lys-Glu-Pro-Met-Thr-Ser-Ala-NH 2 -Leu-Lys-Cys-Tyr-Thr-Cys-Lys-Glu-Pro-Met-Thr-Ser-Ala-
Ser-Cys-Arg-Thr-Ile-Thr-Arg-Cys-Lys-Pro-Glu-Asp-Thr-Ala-Ser-Cys-Arg-Thr-Ile-Thr-Arg-Cys-Lys-Pro-Glu-Asp-Thr-Ala-
Cys-Met-Thr-Thr-Leu-Val-Thr-Val-Glu-Ala-Glu-Tyr-Pro-Phe-Cys-Met-Thr-Thr-Leu-Val-Thr-Val-Glu-Ala-Glu-Tyr-Pro-Phe-
Asn-Gln-Ser- ro-Val -Val-Thr-Arg-Ser-Cys-Ser-Ser-Ser-Cys-Asn-Gln-Ser- ro-Val -Val-Thr-Arg-Ser-Cys-Ser-Ser-Ser-Cys-
Val-Ala-Thr-Asp-Pro-Asp-Ser-Ile-Gly-Ala-Ala-His-Leu-Ile-Val-Ala-Thr-Asp-Pro-Asp-Ser-Ile-Gly-Ala-Ala-His-Leu-Ile-
Phe-Cys-Cys-Phe-Arg-Asp-Leu-Cys-Asn-Ser-Glu-Leu-COOHPhe-Cys-Cys-Phe-Arg-Asp-Leu-Cys-Asn-Ser-Glu-Leu-COOH
(LUS-I)(LUS-I)
und seine cyclischen, glykosylierten, phosphorylierten, acetylierten, amidierten und/oder Verknüpfungen von Seitenketten enthaltenden Derivate sowie Fragmente mit der biologischen Aktivität von LUS-I.and its cyclic, glycosylated, phosphorylated, acetylated, amidated and / or linkages of side chain-containing derivatives and fragments with the biological activity of LUS-I.
2. Verfahren zur Herstellung des Proteins gemäß Anspruch 1 durch Aufreinigung aus humanem Blutfiltrat oder Urin, durch Festphasenpeptidsynthese oder rekombinante Expression in Mikroorganismen, gegebenenfalls modifiziert durch chemische oder biochemische Methoden.2. A process for the production of the protein according to claim 1 by purification from human blood filtrate or urine, by solid phase peptide synthesis or recombinant expression in microorganisms, optionally modified by chemical or biochemical methods.
3. Nukleinsäure dadurch gekennzeichnet, daß sie für das Protein gemäß Anspruch 1 kodiert .3. Nucleic acid, characterized in that it codes for the protein according to claim 1.
4. Antisensenukleotid, dadurch gekennzeichnet, daß es unter stringenten Bedingungen an eine Nukleinsäure gemäß Anspruch 3 bindet .4. antisense nucleotide, characterized in that it binds to a nucleic acid according to claim 3 under stringent conditions.
5. Antikörper, dadurch gekennzeichnet, daß er an ein Protein gemäß Anspruch 1 bindet .5. Antibody, characterized in that it binds to a protein according to claim 1.
6. Inhibitor, dadurch gekennzeichnet, daß er die biologische Aktivität des Proteins gemäß Anspruch 1 hemmt.
6. Inhibitor, characterized in that it inhibits the biological activity of the protein according to claim 1.
7. Inhibitor, dadurch gekennzeichnet, daß er die Expression von LUS-I hemmt .7. Inhibitor, characterized in that it inhibits the expression of LUS-I.
8. Verwendung des Proteins gemäß Anspruch 1, Nukleinsäuren gemäß Anspruch 3, Antisensenukleotide gemäß Anspruch 4, Antikörper gemäß Anspruch 5, Inhibitoren gemäß Anspruch 6 und/oder 7 zur Herstellung eines Arzneimittels zur Behandlung von bakteriellen und viralen Effekten, der Überoder Unterexpression von LUS-I, von Krebserkrankungen, wie Krebserkrankungen des Gebärmutterhalses, das kleinzellige Bronchialkarzinom, Pankreaskarzinom, Mammakarzinom und Melanome sowie von Autoimmunerkrankungen, angioneurotis- chem Ödem, Asthma bronchiale und paroxysmale nächtliche Hämoglobinurie .8. Use of the protein according to claim 1, nucleic acids according to claim 3, antisense nucleotides according to claim 4, antibodies according to claim 5, inhibitors according to claim 6 and / or 7 for the manufacture of a medicament for the treatment of bacterial and viral effects, the over or under expression of LUS I, cancer, such as cancer of the cervix, small cell bronchial carcinoma, pancreatic carcinoma, breast carcinoma and melanoma, as well as autoimmune diseases, angioneurotic edema, bronchial asthma and paroxysmal nocturnal hemoglobinuria.
9. Arzneimittel enthaltend das Protein gemäß Anspruch 1, Nukleinsäuren gemäß Anspruch 3, Antisensenukleotide gemäß Anspruch 4, Antikörper gemäß Anspruch 5, Inhibitoren gemäß Anspruch 6 und/oder 7.9. Medicament containing the protein according to claim 1, nucleic acids according to claim 3, antisense nucleotides according to claim 4, antibodies according to claim 5, inhibitors according to claim 6 and / or 7.
10. Verwendung der Nukleinsäure gemäß Anspruch 3 zur Herstellung eines Medikaments zur Behandlung somatischer oder nicht-somatischer Generkrankung.10. Use of the nucleic acid according to claim 3 for the manufacture of a medicament for the treatment of somatic or non-somatic genetic disease.
11. Transgenes Säugetier mit Gendefizienz für LUS-I.11. Transgenic mammal with gene deficiency for LUS-I.
12. Diagnostikmittel enthaltend des Protein gemäß Anspruch 1, eine Nukleinsäure gemäß Anspruch 3, ein Antisensenukleotid gemäß Anspruch 4, einen Antikörper gemäß Anspruch 5, einen Inhibitor gemäß Anspruch 6 und/oder 7.
12. Diagnostic agent containing the protein according to claim 1, a nucleic acid according to claim 3, an antisense nucleotide according to claim 4, an antibody according to claim 5, an inhibitor according to claim 6 and / or 7.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU85366/98A AU8536698A (en) | 1997-06-09 | 1998-06-09 | Lus-1 human protein, its production and use |
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19724301.0 | 1997-06-09 | ||
| DE19724301 | 1997-06-09 | ||
| DE19749073.5 | 1997-11-06 | ||
| DE1997149073 DE19749073C1 (en) | 1997-11-06 | 1997-11-06 | New protein, LUS-I, and related nucleic acid, antibodies, inhibitors and transgenic animals |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| WO1998056810A2 true WO1998056810A2 (en) | 1998-12-17 |
| WO1998056810A3 WO1998056810A3 (en) | 1999-03-11 |
Family
ID=26037290
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/EP1998/003460 WO1998056810A2 (en) | 1997-06-09 | 1998-06-09 | Lus-1 human protein, its production and use |
Country Status (2)
| Country | Link |
|---|---|
| AU (1) | AU8536698A (en) |
| WO (1) | WO1998056810A2 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004091646A3 (en) * | 2003-04-16 | 2004-12-16 | Univ Lausanne | Use of slurp-1 for treating diseases related to acetylcholine receptors dysfunction |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5298604A (en) * | 1992-07-27 | 1994-03-29 | Sloane Nathan H | Parital primary amino acid sequence of the antineoplastic protein (ANUP); a cytokine present in granulocytes |
| WO1994014959A1 (en) * | 1992-12-22 | 1994-07-07 | Applied Research Systems Ars Holding N.V. | New protein from urine named component b |
-
1998
- 1998-06-09 AU AU85366/98A patent/AU8536698A/en not_active Abandoned
- 1998-06-09 WO PCT/EP1998/003460 patent/WO1998056810A2/en active Application Filing
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5298604A (en) * | 1992-07-27 | 1994-03-29 | Sloane Nathan H | Parital primary amino acid sequence of the antineoplastic protein (ANUP); a cytokine present in granulocytes |
| WO1994014959A1 (en) * | 1992-12-22 | 1994-07-07 | Applied Research Systems Ars Holding N.V. | New protein from urine named component b |
Non-Patent Citations (2)
| Title |
|---|
| MASTRANGELI R.: "H. sapiens ARS gene, component B, ARS" EMBL DATABASE,11. September 1996, XP002087799 HEIDELBERG, DE * |
| SLOANE N.H. ET AL.: "Studies on an antineoplastic fraction from human urine characterization of the major protein in this fraction" BIOCHEMICAL JOURNAL, Bd. 234, Nr. 2, 1986, Seiten 355-362, XP002087807 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004091646A3 (en) * | 2003-04-16 | 2004-12-16 | Univ Lausanne | Use of slurp-1 for treating diseases related to acetylcholine receptors dysfunction |
| JP2006523678A (en) * | 2003-04-16 | 2006-10-19 | アプライド リサーチ システムズ エーアールエス ホールディング ナームロゼ フェンノートシャップ | SLURP-1 compositions and methods of using the same |
| US7135454B2 (en) | 2003-04-16 | 2006-11-14 | Applied Research Systems Ars Holding N.V. | Use of SLURP-1 compositions for treating schizophrenia |
| AU2004229237B2 (en) * | 2003-04-16 | 2009-06-04 | Merck Serono Sa | Use of SLURP-1 for treating diseases related to acetylcholine receptors dysfunction |
| US7691808B2 (en) | 2003-04-16 | 2010-04-06 | Merck Serono Sa | Use of Slurp-1 for treating diseases related to acetylcholine receptor dysfunction |
| NO337494B1 (en) * | 2003-04-16 | 2016-04-25 | Merck Serono Sa | Compositions comprising an effective amount of Slurp-1 for use in the treatment of neurological disorders and skin disorders |
Also Published As
| Publication number | Publication date |
|---|---|
| AU8536698A (en) | 1998-12-30 |
| WO1998056810A3 (en) | 1999-03-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP1027440B1 (en) | Inhibitor protein of the wnt signal pathway | |
| KR910002692B1 (en) | Fsh | |
| Terry et al. | The cDNA sequence coding for prepro-PGS (prepro-magainins) and aspects of the processing of this prepro-polypeptide. | |
| JP2682858B2 (en) | Leukemia inhibitory factor | |
| DE19757250A1 (en) | Insulin-like growth factor binding protein and its use | |
| JPH05500211A (en) | Megakaryocytic factor | |
| DE69530944T2 (en) | KERATINOCYTE GROWTH FACTOR 2 | |
| EP0736095B1 (en) | Human circulating cytokine cc-1 | |
| EP3530667B1 (en) | Peptide having anti-obesity and anti-diabetes efficacy and use thereof | |
| WO1998056810A2 (en) | Lus-1 human protein, its production and use | |
| WO2001034640A2 (en) | Peptide (virip) which inhibits a circulating virus in humans and the use thereof | |
| DE10027383A1 (en) | Nucleic acid molecule comprising a nucleic acid sequence coding for a chemokine, a neuropeptide precursor or at least one neuropeptide | |
| JPH04500603A (en) | Cloned nephritis antigen | |
| KR101025352B1 (en) | Physiologically active complex | |
| US5440022A (en) | Hepatokine and methods for its use | |
| AU783373B2 (en) | Epithelial cell growth inhibitors | |
| EP0612349B1 (en) | Novel thrombin inhibitor protein from reduvii | |
| DE69832447T2 (en) | Gene encoding Afadin-1 | |
| EP0609242B1 (en) | New thrombin-inhibiting protein isolated from ticks | |
| US20040115191A1 (en) | Method for treating psoriasis | |
| DE19749073C1 (en) | New protein, LUS-I, and related nucleic acid, antibodies, inhibitors and transgenic animals | |
| DE69836226T2 (en) | IFN RECEPTOR 1 BINDING PROTEINS, DNA PROCESSING THEREFOR AND METHOD FOR REGULATING THE CELLULAR ANSWER TO INTERFERONE | |
| DE19508672A1 (en) | Cyclic parathyroid hormone fragments with lactam bridge | |
| EP1161535B1 (en) | Rna polymerase i transcription factor tif-ia | |
| WO1994019462A1 (en) | Harp family growth factors, preparation methods therefor and uses thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| AK | Designated states |
Kind code of ref document: A2 Designated state(s): AL AU BA BB BG BR CA CN CU CZ EE GE GW HU ID IL IS JP KP KR LC LK LR LT LV MG MK MN MX NO NZ PL RO SG SI SK SL TR TT UA US UZ VN YU |
|
| AL | Designated countries for regional patents |
Kind code of ref document: A2 Designated state(s): GH GM KE LS MW SD SZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN ML MR NE SN TD TG |
|
| DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
| AK | Designated states |
Kind code of ref document: A3 Designated state(s): AL AU BA BB BG BR CA CN CU CZ EE GE GW HU ID IL IS JP KP KR LC LK LR LT LV MG MK MN MX NO NZ PL RO SG SI SK SL TR TT UA GH |
|
| AL | Designated countries for regional patents |
Kind code of ref document: A3 Designated state(s): GH GM KE LS MW SD SZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG |
|
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
| NENP | Non-entry into the national phase |
Ref country code: JP Ref document number: 1999501578 Format of ref document f/p: F |
|
| 122 | Ep: pct application non-entry in european phase | ||
| NENP | Non-entry into the national phase |
Ref country code: CA |