WO1999062933A2 - Peptides conçus de maniere rationnelle, leur production et leur utilisation - Google Patents

Peptides conçus de maniere rationnelle, leur production et leur utilisation Download PDF

Info

Publication number
WO1999062933A2
WO1999062933A2 PCT/DE1999/001575 DE9901575W WO9962933A2 WO 1999062933 A2 WO1999062933 A2 WO 1999062933A2 DE 9901575 W DE9901575 W DE 9901575W WO 9962933 A2 WO9962933 A2 WO 9962933A2
Authority
WO
WIPO (PCT)
Prior art keywords
peptides
peptide
autoantibodies
binding
activity
Prior art date
Application number
PCT/DE1999/001575
Other languages
German (de)
English (en)
Other versions
WO1999062933A3 (fr
Inventor
Gerd Wallukat
Gisbert Schneider
Wieland SCHRÖDL
Johannes Müller
Wolfgang RÖNSPECK
Paul Wrede
Rudolf Kunze
Original Assignee
Affina Immuntechnik 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
Priority claimed from DE19826442A external-priority patent/DE19826442A1/de
Application filed by Affina Immuntechnik Gmbh filed Critical Affina Immuntechnik Gmbh
Priority to EP99936345A priority Critical patent/EP1082339A2/fr
Priority to AU51504/99A priority patent/AU5150499A/en
Publication of WO1999062933A2 publication Critical patent/WO1999062933A2/fr
Publication of WO1999062933A3 publication Critical patent/WO1999062933A3/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
    • C07K1/047—Simultaneous synthesis of different peptide species; Peptide libraries

Definitions

  • the invention relates to rationally designed peptides, their preparation and their use. Areas of application of the invention are medicine and the pharmaceutical industry.
  • DCM Dilated cardiomyopathy
  • *% AK + proportion of antibody-positive patients with the respective clinical diagnosis.
  • the procedure known as immunapheresis, removes all of the immunoglobulin from the body or the bloodstream.
  • target-specific autoantibodies only small portions of the body's immunoglobulin, so-called target-specific autoantibodies, could often be attributed to the cause of the disease.
  • DCM dilated cardiomyopathy
  • the pathological autoantibodies are detected in a microscopic biological test on cultivated, neonatal, spontaneously contracting rat cardiac muscle cells. Their beat frequency increases in the presence of certain ß1 receptor-specific autoantibodies.
  • the aim of the invention is to provide new peptides suitable for the binding and removal of pathogenic autoantibodies. It is based on the task of developing an effective method for generating such peptides.
  • the invention is implemented according to the claims, the subclaims are preferred variants.
  • the method according to the invention for generating peptides which are suitable for binding autoantibodies is characterized by the following steps:
  • the physicochemical space which is the size of the number of amino acids of the peptide, represents not only the sequences "around the seed peptide", but the entire sequence space.
  • DCM dilated cardiomyopathy
  • the amino acid sequences of the epitopes recognized by autoantibodies on the corresponding receptors are known in detail at DCM.
  • the natural sequences of the relevant receptor areas are used in order to use them after the solid phase coupling for the elimination of the autoantibodies from the blood plasma.
  • the natural amino acid sequence (10 amino acids; pos. 107-116: ARRC YNDPKC) or partial sequences thereof from Loop2 of the ⁇ 1-adrenergic receptor was selected as the original or seed peptide.
  • This epitope contains the autoimmunologically relevant structure. It is known to bind autoantibodies and if this peptide is added to an appropriate biological test system, the autoantibodies present in the test system can be neutralized. This peptide is also coupled in a solid phase, ie it recognizes and binds the corresponding ß1-adrenergic autoantibodies from immunoglobulin fractions from patients.
  • the natural peptide does not fully fulfill this task and has a low neutralizing capacity of the autoantibodies in the biological test.
  • the peptide from the Loop2 region of the ⁇ 1-adrenergic human receptor was the starting point for the search strategy to find a peptide with higher antibody binding efficiency.
  • an amino acid sequence was identified that does not occur in nature or has never been described in the corresponding databases.
  • variants of peptides were developed with the help of a special computer-aided computation method, which obey certain physicochemical laws and accordingly have different binding properties towards autoantibodies and immunoglobulins.
  • the peptide DRFGDKDIAF was identified, which no longer has any sequence similarity to the natural epitope (Loop2) and efficiently neutralizes the effect of the autoantibodies on the pulsation frequency of the heart muscle cells in the cell biological test used. With the help of this peptide, the binding and elimination of the Loop2-specific, ßl-adrenergic autoantibodies succeed.
  • a certain number of sequence variants (DCM project: 90 variants) is generated using a defined algorithm: each amino acid residue is described with suitable physicochemical parameters.
  • DCM seed peptide these were the properties 'volume' (1) and 'hydrophobicity' (libophobia) (2).
  • the variants of the seed peptide to be produced are now preferably taken from the closer environment of the seed peptide in the chemical space, but also include some peptides far from the seed peptide.
  • sequences were chosen so that their distances from the seed peptide in chemical space approximated a Gaussian distribution (or other localized distribution).
  • the rational approach here is that peptides with a similar or even higher activity (effect, property) can be expected in the immediate vicinity of the seed peptide in the chemical space (3).
  • a Gaussian distribution of peptides is very important for the next computer-aided design step, the development of sequence-activity relationships.
  • the new peptides generated in step 1 are examined for their activity (action, desired property) using a suitable test method.
  • Figure 1 shows the result: As the distance (Euclidean distance measure) from the seed peptide increases, the average activity of the peptide variants decreases. Two peculiarities can be found: on the one hand the average activity of the peptides closely adjacent to the seed peptide is above that of the seed peptide, and on the other hand there are significant activities far away from the seed peptide. This observation confirms the knowledge-based approach and the method for the systematic generation of peptide libraries: Several non-natural peptides with an increased ability to bind antibodies compared to the seed peptide were found.
  • sequence-activity relationships are modeled.
  • artificial neural networks are used for this.
  • the peptides generated in step 1 and tested in step 2 together with the seed peptide are numerically described with suitable physicochemical parameters (see step 1).
  • suitable physicochemical parameters see step 1.
  • a mathematical connection is established between the description of the properties of the peptides and their measured activity. This is an optimization process that is described in detail in the literature (4,5).
  • the consideration of both peptides with high activity and inactive peptides in this process is necessary for a meaningful modeling.
  • the defined selection of peptides in step 1 fulfills this requirement.
  • the neural network optimized in step 3 is used as a quality function for the computer-aided systematic search in the sequence space (amount of all possible peptides of a given one Length).
  • An evolutionary algorithm is used as a search strategy (this method is described in the literature (6)). The principle is as follows: In an iterative process, new variants of a parent peptide are generated by the computer program, then all variants are evaluated with regard to their ability to bind antibodies using the neural network (see step 3), and the best peptide variant (s) ) are selected as parent peptides for the next cycle.
  • the DCM project we received the sequence DRFGDKDIAF as the output of the computer program. The neural network calculated the highest activity for this peptide.
  • the sequence has an identity with the original seed peptide (see step 1) in only two positions (arginine-2, aspartic acid-7). 73% of all potentially strongly antibody-binding peptides have the Asp-7 residue according to the prediction of the neural network, 17% have an asparagine in position 7.
  • the system can also be used to generate peptides that do not have a certain property, therefore do not bind antibodies.
  • the sequence FVRRTYYPER was proposed as such by the computer system.
  • Test systems especially bioassays:
  • the peptides designed by the computer in step 4 are experimentally examined for their actual biochemical biological effect.
  • this test consisted of measuring the beat rate of embryonic rat myocytes with and without adding peptides to the medium.
  • the normal rate of increase of human antibodies against the ⁇ 1-adrenergic receptor was in the medium.
  • the exact test procedure for the DCM project is described in the literature (7).
  • the result corresponded to the expectations and predictions of the computer program:
  • the peptide DRFGDKDIAF was active, ie the rate of heart muscle cells increased by the pathogenic antibodies was depressed to the normal frequency, and the peptide FVRRTYYPER, which was designed as inactive, had no measurable effect (Figure 2) .
  • the peptide DRFGDKDIAF had more pronounced effects than the natural antigen of the ⁇ 1-adrenergic receptor at all peptide concentrations used (10 ⁇ g / ml, 1 ⁇ g / ml, 0.1 ⁇ g / ml).
  • the natural antigen had only a very weak effect (only 10% of the normal activity of the myocardial cells was observed), while the peptide designed according to the invention was able to restore 50% of the normal activity of the myocardial cells.
  • Literature :

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne des peptides conçus de manière rationnelle, leur production et leur utilisation à des fins biotechnologiques, médicales, diagnostiques et thérapeutiques. Selon l'invention, le procédé de production de peptides adaptés à la liaison d'auto-anticorps est caractérisé par les étapes suivantes: identification d'un peptide de semence présentant une activité appropriée; production d'une sélection restreinte de variantes de peptides à partir de la zone physico-chimique autour du peptide de semence, au moyen d'une méthode de calcul assistée par ordinateur; synthèse et test de l'activité de ces variantes de peptides; modélisation d'un rapport quantitatif séquence/activité au moyen d'un réseau neuronal artificiel; et production de peptides par évolution simulée dans la zone de séquence au moyen du réseau neuronal ayant subi un apprentissage. L'invention concerne en particulier le peptide DRFGDKDIAF.
PCT/DE1999/001575 1998-06-02 1999-05-28 Peptides conçus de maniere rationnelle, leur production et leur utilisation WO1999062933A2 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP99936345A EP1082339A2 (fr) 1998-06-02 1999-05-28 Peptides con us de maniere rationnelle, leur production et leur utilisation
AU51504/99A AU5150499A (en) 1998-06-02 1999-05-28 Rationally designed peptides, production and use thereof

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
DE19824641 1998-06-02
DE19824641.2 1998-06-02
DE19826442A DE19826442A1 (de) 1998-06-02 1998-06-13 Rational designte Peptide, ihre Herstellung und ihre Verwendung
DE19826442.9 1998-06-13

Publications (2)

Publication Number Publication Date
WO1999062933A2 true WO1999062933A2 (fr) 1999-12-09
WO1999062933A3 WO1999062933A3 (fr) 2000-04-06

Family

ID=26046590

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DE1999/001575 WO1999062933A2 (fr) 1998-06-02 1999-05-28 Peptides conçus de maniere rationnelle, leur production et leur utilisation

Country Status (3)

Country Link
EP (1) EP1082339A2 (fr)
AU (1) AU5150499A (fr)
WO (1) WO1999062933A2 (fr)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002016431A2 (fr) * 2000-08-24 2002-02-28 Max-Delbrück-Centrum für Molekulare Medizin PEPTIDES DU RECEPTEUR α1-ADRENERGIQUE ET LEUR UTILISATION POUR TRAITER LE PSORIASIS
WO2002020564A2 (fr) * 2000-09-05 2002-03-14 Callistogen Ag Procede servant a identifier des sequences de peptides possedant une fonctionnalite specifique
EP1832600A1 (fr) * 2006-03-09 2007-09-12 Max-Delbrück-Centrum Für Molekulare Medizin Peptides contre autoanticorps associees du glaucome et leur utilisation.
EP2172481A1 (fr) 2008-10-06 2010-04-07 Novoplant GmbH Formats d'anticorps stables de manière protéolytique
EP2199305A1 (fr) 2008-12-18 2010-06-23 Max-Delbrück-Centrum Peptides contre les anticorps associés au CRPS et utilisation de ces peptides
WO2011020529A2 (fr) 2009-08-19 2011-02-24 Merck Patent Gmbh Anticorps pour la détection de complexes d'intégrine dans une matière ffpe
WO2013023852A1 (fr) 2011-08-12 2013-02-21 E.R.D.E.-Aak-Diagnostik Gmbh Auto-anticorps agonistes contre le récepteur adrénergique alpha1 et le récepteur adrénergique bêta2 dans la démence d'alzheimer et vasculaire

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998023781A1 (fr) * 1996-11-26 1998-06-04 Johns Hopkins University Systeme de detection de ligands et procedes d'utilisation associes
WO1998053407A1 (fr) * 1997-05-23 1998-11-26 The Scripps Research Institute Prediction de motifs de liaison relatifs de peptides biologiquement actifs et de mimetiques de peptides

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998023781A1 (fr) * 1996-11-26 1998-06-04 Johns Hopkins University Systeme de detection de ligands et procedes d'utilisation associes
WO1998053407A1 (fr) * 1997-05-23 1998-11-26 The Scripps Research Institute Prediction de motifs de liaison relatifs de peptides biologiquement actifs et de mimetiques de peptides

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
G SCHNEIDER ET AL.: "Peptide design by artificial neural networks and computer-based evolutionary search" PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA., Bd. 95, Nr. 21, 13. Oktober 1998 (1998-10-13), Seiten 12179-12184, XP002127398 NATIONAL ACADEMY OF SCIENCE. WASHINGTON., US ISSN: 0027-8424 *
J SCHUCHHARDT ET AL.: "Local structural motifs of protein backbones are classified by self-organizing neural networks" PROTEIN ENGINEERING., Bd. 9, Nr. 10, Oktober 1996 (1996-10), Seiten 833-842, XP002127396 OXFORD UNIVERSITY PRESS, SURREY., GB ISSN: 0269-2139 *
P WREDE ET AL.: "Peptide design aided by neural networks: biological activity of rtificial signal peptidase I cleavage sites " BIOCHEMISTRY., Bd. 37, Nr. 11, 17. M{rz 1998 (1998-03-17), Seiten 3588-3593, XP002127397 AMERICAN CHEMICAL SOCIETY. EASTON, PA., US ISSN: 0006-2960 *

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002016431A2 (fr) * 2000-08-24 2002-02-28 Max-Delbrück-Centrum für Molekulare Medizin PEPTIDES DU RECEPTEUR α1-ADRENERGIQUE ET LEUR UTILISATION POUR TRAITER LE PSORIASIS
WO2002016431A3 (fr) * 2000-08-24 2002-07-18 Max Delbrueck Centrum PEPTIDES DU RECEPTEUR α1-ADRENERGIQUE ET LEUR UTILISATION POUR TRAITER LE PSORIASIS
WO2002020564A2 (fr) * 2000-09-05 2002-03-14 Callistogen Ag Procede servant a identifier des sequences de peptides possedant une fonctionnalite specifique
WO2002020564A3 (fr) * 2000-09-05 2003-10-02 Callistogen Ag Procede servant a identifier des sequences de peptides possedant une fonctionnalite specifique
EP1832600A1 (fr) * 2006-03-09 2007-09-12 Max-Delbrück-Centrum Für Molekulare Medizin Peptides contre autoanticorps associees du glaucome et leur utilisation.
WO2007101732A1 (fr) 2006-03-09 2007-09-13 Max-Delbrück-Centrum für Molekulare Medizin Peptides dirigés contre des auto-anticorps associés au glaucome et utilisation de ces peptides
EP2172481A1 (fr) 2008-10-06 2010-04-07 Novoplant GmbH Formats d'anticorps stables de manière protéolytique
EP2199305A1 (fr) 2008-12-18 2010-06-23 Max-Delbrück-Centrum Peptides contre les anticorps associés au CRPS et utilisation de ces peptides
WO2010069570A2 (fr) 2008-12-18 2010-06-24 Max-Delbrück-Centrum für Molekulare Medizin Peptides contre des anticorps autologues associés à crps et utilisation de ces peptides
WO2011020529A2 (fr) 2009-08-19 2011-02-24 Merck Patent Gmbh Anticorps pour la détection de complexes d'intégrine dans une matière ffpe
WO2013023852A1 (fr) 2011-08-12 2013-02-21 E.R.D.E.-Aak-Diagnostik Gmbh Auto-anticorps agonistes contre le récepteur adrénergique alpha1 et le récepteur adrénergique bêta2 dans la démence d'alzheimer et vasculaire

Also Published As

Publication number Publication date
AU5150499A (en) 1999-12-20
WO1999062933A3 (fr) 2000-04-06
EP1082339A2 (fr) 2001-03-14

Similar Documents

Publication Publication Date Title
EP0276723B1 (fr) Précurseur protéinique de polypeptide-APC, ADN le codant et utilisation diagnostique de cet ADN et de cette protéine
DE69535133T2 (de) Humanisierte antikörper gegen das leukozytenadhäsionsmolekül vla-4
DE69934967T2 (de) Verfahren zur verminderung der immunogenität von proteinen
EP1896497B1 (fr) Procede pour le diagnostic de maladies rhumatismales
DE69434223T2 (de) In der Behandlung von IL4 auslösenden Krankheiten nützliche rekombinante IL4-Antikörper
DE69836129T2 (de) Oxidierte Fragmente von Apolipoprotein B und deren Verwendung
DE60017019T2 (de) Antikörper gegen Spaltprodukte von Vimentin
EP1451224A2 (fr) Peptide ou proteine contenant une boucle c'-d de la famille de recepteurs cd28
WO1999062933A2 (fr) Peptides conçus de maniere rationnelle, leur production et leur utilisation
DE19826442A1 (de) Rational designte Peptide, ihre Herstellung und ihre Verwendung
EP0955361A1 (fr) Anticorps monoclonaux contre des antigènes associés aux tumeurs, procédés de préparation et utilisation
DE69333709T2 (de) T-zell epitope des raygraspollen-allergens
DE69434665T2 (de) Ein hausmilbeallergen (der p iii), dessen anwendung und dafür kodierte nukleinsäuren
EP0696597B1 (fr) Anticorps monoclonaux pour la détermination immunologique sélective des formes de la laminine intacte à haut poids moléculaire dans des fluides corporels
DE69836679T2 (de) Peptide zur behandlung des systemischen lupus erythematosus
WO2005112985A2 (fr) Agents et procedes pour le diagnostic, la prophylaxie et la therapie de maladies du tissu conjonctif
DE4002981A1 (de) Humane, epididymis-spezifische polypeptide und deren verwendung zur therapie und diagnose maennlicher infertilitaet
DE69839163T2 (de) Methode zur Feststellung der therapeutischen Aktivität eines MBP-Fragments
DE69834408T2 (de) Strukturelles Peptid mit antigenischen Eigenschaften, antigenischen Verbindungen und Verwendungen im Nachweis, Vorbeugung und Behandlung einer VHC-Infektion
DE102006012613B4 (de) Verfahren zur Bestimmung der therapeutischen Wirksamkeit von Substanzen
EP0133551A2 (fr) Anticorps monoclonal qui reconnaît une structure commune à l'interleukine-2 humaine (TCGF) et à la chaîne légère lambda de l'immunoglobuline humaine et lignées cellulaires d'hybridome produisant ces anticorps monoclonaux
WO2002073209A2 (fr) Procede d'identification d'epitopes immunoreactifs sur des proteines et utilisation desdits epitopes a des fins prophylactiques et therapeutiques
DE60211714T2 (de) Isolierung membrangebundener ligandenspezifischer komplexe
DE19905048A1 (de) Verwendung depletorisch oder mitogen wirkender Antikörper in der Immuntherapie
WO2002016431A2 (fr) PEPTIDES DU RECEPTEUR α1-ADRENERGIQUE ET LEUR UTILISATION POUR TRAITER LE PSORIASIS

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AL AM AU AZ BA BB BG BR BY CA CN CU CZ EE GE GH GM HR HU ID IL IS JP KE KG KP KR KZ LC LK LR LS LT LV MD MG MK MN MW MX NO NZ PL RO RU SD SG SI SK SL TJ TM TR TT UA UG US UZ VN YU ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE

121 Ep: the epo has been informed by wipo that ep was designated in this application
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 AM AU AZ BA BB BG BR BY CA CN CU CZ EE GE GH GM HR HU ID IL IS JP KE KG KP KR KZ LC LK LR LS LT LV MD MG MK MN MW MX NO NZ PL RO RU SD SG SI SK SL TJ TM TR TT UA UG US UZ VN YU ZW

AL Designated countries for regional patents

Kind code of ref document: A3

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE

WWE Wipo information: entry into national phase

Ref document number: 1999936345

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1999936345

Country of ref document: EP

NENP Non-entry into the national phase

Ref country code: CA

WWW Wipo information: withdrawn in national office

Ref document number: 1999936345

Country of ref document: EP