EP0411096A1 - Sequences d'adn codant pour des variantes de pth, variantes de pth, vecteur d'expression, hote bacterien, utilisation et composition therapeutique - Google Patents

Sequences d'adn codant pour des variantes de pth, variantes de pth, vecteur d'expression, hote bacterien, utilisation et composition therapeutique

Info

Publication number
EP0411096A1
EP0411096A1 EP90903352A EP90903352A EP0411096A1 EP 0411096 A1 EP0411096 A1 EP 0411096A1 EP 90903352 A EP90903352 A EP 90903352A EP 90903352 A EP90903352 A EP 90903352A EP 0411096 A1 EP0411096 A1 EP 0411096A1
Authority
EP
European Patent Office
Prior art keywords
amino acids
terminal
dna sequence
variant
pth
Prior art date
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
EP90903352A
Other languages
German (de)
English (en)
Inventor
Edgar Wingender
Heiko Mielke
Gisela Bercz
Klaus-Dieter Schlüter
Hubert Mayer
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
Helmholtz Zentrum fuer Infektionsforschung HZI GmbH
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.)
Filing date
Publication date
Application filed by Helmholtz Zentrum fuer Infektionsforschung HZI GmbH filed Critical Helmholtz Zentrum fuer Infektionsforschung HZI GmbH
Publication of EP0411096A1 publication Critical patent/EP0411096A1/fr
Ceased legal-status Critical Current

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
    • C07K14/575—Hormones
    • C07K14/635—Parathyroid hormone, i.e. parathormone; Parathyroid hormone-related peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • 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
    • C07K2319/61—Fusion polypeptide containing an enzyme fusion for detection (lacZ, luciferase)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C07K2319/74—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor
    • C07K2319/75—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones

Definitions

  • Parathyroid hormone regulates the calcium level of the blood plasma by u. a. Develops and dismantles effects on the skeleton, but also increases calcium reabsorption by the kidney. There are indications that the bone-building effect of PTH can be used for the clinical treatment of osteoporosis; see. J. Bone Minera l Res., 1 (1987) 377-382.
  • a DNA sequence which encodes a variant of PTH (1-84) in which
  • Amino acids is shortened, the totality of the amino acids encoded by the DNA sequence not being arranged in an order that occurs in bPTH, pPTH, rPTH, cPTH or hPTH.
  • a DNA sequence which encodes a variant of PTH (1-84) in which
  • Amino acids are exchanged for another amino acid, where
  • the N-terminal (position +1) is extended by one to two amino acids and, if necessary, the C-terminal (position +84) is shortened by one or more amino acids or
  • N-terminal (position +1) and / or C-terminal (position +84) is shortened by one or more amino acids, the totality of the amino acids coded by the DNA sequence not being arranged in an order which is shown in bPTH, pPTH, rPTH, cPTH or hPTH occurs.
  • a DNA sequence which encodes a variant of PTH (1-84) in which
  • Amino acids are exchanged for another amino acid, except for tyrosine at position 34 and further
  • the N-terminal (position +1) is extended by one to two amino acids and, if necessary, the C-terminal (position +84) is shortened by one or more amino acids or
  • N-terminal (position +1) and / or C-terminal (position +84) is shortened by one or more amino acids, the totality of the amino acids coded by the DNA sequence not being arranged in an order which is shown in bPTH, pPTH, rPTH, cPTH or hPTH occurs.
  • a DNA sequence is provided which encodes a variant of PTH (1-84) in which
  • Amino acids are exchanged for another amino acid, where
  • N-terminal (position +1) is extended by one or two amino acids and, if necessary, the C-terminal (position +84) is shortened by one or more amino acids or
  • N-terminal (position +1) and / or C-terminal (position +84) is shortened by one or more amino acids, the totality of the amino acids coded by the DNA sequence not being arranged in an order which is shown in bPTH, pPTH, rPTH, cPTH or hPTH occurs.
  • a DNA sequence which encodes a variant of PTH (1-84) in which
  • the N-terminal (position +1) is extended by one to two amino acids and, if necessary, the C-terminal (position +84) is shortened by one or more amino acids or
  • - N-terminal (position +1) and / or C-terminal (position +84) is possibly shortened by one or more amino acids, where
  • the entirety of the amino acids encoded by the DNA sequence is not arranged in an order that occurs in bPTH, pPTH, rPTH, cPTH or hPTH.
  • the DNA sequence preferably encodes a PTH variant whose N-terminus is extended by proline. Also preferred are hPTH, bPTH, pPTH, rPTH or cPTH variants.
  • 1 denotes the first nucleotide of the first PTH codon, so that the numbers to be taken from the following Table 1 each indicate the position of the first nucleotide shown in relation to the first nucleotide of the first PTH codon.
  • the invention also relates to PTH variants which are encoded by one of the DNA sequences described above, and in particular to variants of PTH from human (h), from bovine (b), from
  • the PTH variants according to the invention can be formally divided into the following four groups.
  • Group 1 conservative amino acid changes in positions 27 and 29, which stimulate both adenylate cyclase and DNA synthesis ("functional PTH equivalents"); Examples: Q29N; K27R.
  • Group 2 non-conservative amino acid exchanges (variations under charge reversal or charge deletion, change in hydrogen bonds, reversal of hydrophobicity) in the ranges 29 to 31 or 33 to 34 as well as exchanges in position 32, which lead to a significant reduction in the stimulation of the DNA Synthesis and largely maintain the effect on adenylate cyclase; Examples: Q29A, H32L, H32K, H32R.
  • Group 3 non-conservative amino acid exchanges (as defined above) in the range 1 to 27, which lead to a significant reduction in adenylate cyclase stimulation and partially increase or leave the DNA synthesis unaffected.
  • Group 4 non-conservative amino acid exchanges in the range 1 to 27, which receive stimulation of adenylate cyclase but reduce DNA synthesis; Example: L11K.
  • the invention relates to hybrid peptides which comprise a PTH variant according to the invention, expression vectors for the expression of the proteins according to the invention, bacterial hosts such as E. coli for the expression vectors, the use of the DNA sequences for the production of the peptides according to the invention and therapeutic compositions which use them Contain peptides.
  • Relevant prior art are DE-A-37 25 319.0 and DE-A-37 25 320.4.
  • the expression plasmid pEX-pPTH from DE-A-37 25 319.0 was used. A 123 base pair BamHI / SstI fragment of this plasmid was cloned into the phage vector Ml3mpl8. The mutants listed in Table 1 were thus produced with the aid of synthetic oligonucleotides according to the gapped duplex method (Methods Enzymol., 100 (1983) 468-500).
  • the PTH variants thus obtained were tested for their ability in an in vitro test according to Mohr and Hesch (Biochem. J., 188 (1980) 649-656) to stimulate adenylate cyclase in isolated membranes of adrenal cortex of the pig.
  • the PTH genes mentioned can be produced using peptide synthesis machines.
  • the underlined bases are different from the recombinant wild type.
  • Amino acids refer to the first amino acid of human parathyroid hormone (Ser (1)).
  • Tab. 2 Relative activity of the PTH variants on renal

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Endocrinology (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pharmacology & Pharmacy (AREA)
  • General Chemical & Material Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Public Health (AREA)
  • Engineering & Computer Science (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Toxicology (AREA)
  • Zoology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Obesity (AREA)
  • Hematology (AREA)
  • Diabetes (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne des séquences d'ADN codant pour des variantes de PTH, ces variantes, des vecteurs d'expression, des hôtes bactériens, leurs applications et compositions thérapeutiques.
EP90903352A 1989-02-23 1990-02-23 Sequences d'adn codant pour des variantes de pth, variantes de pth, vecteur d'expression, hote bacterien, utilisation et composition therapeutique Ceased EP0411096A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE3905606 1989-02-23
DE3905606 1989-02-23

Publications (1)

Publication Number Publication Date
EP0411096A1 true EP0411096A1 (fr) 1991-02-06

Family

ID=6374759

Family Applications (1)

Application Number Title Priority Date Filing Date
EP90903352A Ceased EP0411096A1 (fr) 1989-02-23 1990-02-23 Sequences d'adn codant pour des variantes de pth, variantes de pth, vecteur d'expression, hote bacterien, utilisation et composition therapeutique

Country Status (4)

Country Link
US (2) US5457047A (fr)
EP (1) EP0411096A1 (fr)
JP (1) JPH03504201A (fr)
WO (1) WO1990010067A1 (fr)

Families Citing this family (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU674500B2 (en) 1991-11-04 1997-01-02 Genetics Institute, Llc Recombinant bone morphogenetic protein heterodimers, compositions and methods of use
US5382658A (en) * 1992-04-03 1995-01-17 Allelix Biopharmaceuticals Inc. Stability-enhanced variants of parathyroid hormone
US5814603A (en) * 1992-06-12 1998-09-29 Affymax Technologies N.V. Compounds with PTH activity
CA2098639A1 (fr) * 1992-06-19 1993-12-20 K. Anne Kronis Variantes de l'hormone parathyroidienne osteostimulantes et non vasoactives
US5589452A (en) * 1992-07-14 1996-12-31 Syntex (U.S.A.) Inc. Analogs of parathyroid hormone and parathyroid hormone related peptide: synthesis and use for the treatment of osteoporosis
AU672790B2 (en) * 1992-07-15 1996-10-17 Novartis Ag Variants of parathyroid hormone and its fragments
US5969095A (en) * 1995-07-13 1999-10-19 Biomeasure, Inc. Analogs of parathyroid hormone
US5723577A (en) * 1995-07-13 1998-03-03 Biomeasure Inc. Analogs of parathyroid hormone
US5955574A (en) * 1995-07-13 1999-09-21 Societe De Conseils De Recherches Et D'applications Scientifiques, S.A. Analogs of parathyroid hormone
US7410948B2 (en) * 1995-07-13 2008-08-12 Societe De Conseils De Recherches Et D'applications Scientifiques, Sas Analogs of parathyroid hormone
US6544949B1 (en) 1995-07-13 2003-04-08 Societe De Conseils De Recherches Et D'applications Scientifiques, S.A.S. Analogs of parathyroid hormone
US5700774A (en) * 1996-03-26 1997-12-23 Genetics Institute, Inc. Compositions comprising bone morphogenic proteins and truncated parathyroid hormone related peptide, and methods of inducing cartilage by administration of same
ATE357244T1 (de) 2003-09-12 2007-04-15 Wyeth Corp Injizierbare feste calciumphosphat-stäbe zur abgabe von osteogenen proteinen
US20060166251A1 (en) * 2005-01-26 2006-07-27 Archambault Joanne M Use of sFRPs as markers of BMP activity
CA2601436A1 (fr) * 2005-03-30 2006-10-05 Wyeth Methodes pour stimuler la pousse des cheveux par administration de proteines bmp
US8298789B2 (en) * 2007-08-09 2012-10-30 Usv Limited Orthogonal process for purification of recombinant human parathyroid hormone (rhPTH) (1-34)
US20090227503A1 (en) * 2008-02-07 2009-09-10 University Of Rochester Parathyroid hormone treatment for enhanced allograft and tissue-engineered reconstruction of bone defects

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3886132A (en) * 1972-12-21 1975-05-27 Us Health Human parathyroid hormone
US4423037A (en) * 1982-05-13 1983-12-27 The General Hospital Corporation Inhibitors of peptide hormone action
DE3725319A1 (de) * 1987-07-30 1989-02-09 Biotechnolog Forschung Gmbh Pth-modifikationen und therapeutische zusammensetzungen, die sie enthalten und daraus bestehen
DE3725320A1 (de) * 1987-07-30 1989-02-09 Biotechnolog Forschung Gmbh Expressionsvektoren und verfahren unter deren verwendung zur gewinnung von cro/ss-galaktosidase/pth-fusionsproteinen und von pth

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9010067A1 *

Also Published As

Publication number Publication date
JPH03504201A (ja) 1991-09-19
US5457047A (en) 1995-10-10
WO1990010067A1 (fr) 1990-09-07
US5455329A (en) 1995-10-03

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