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WO1999041407A1 - Procede de developpement d'un principe actif pharmaceutique - Google Patents

Procede de developpement d'un principe actif pharmaceutique Download PDF

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Publication number
WO1999041407A1
WO1999041407A1 PCT/EP1999/000876 EP9900876W WO9941407A1 WO 1999041407 A1 WO1999041407 A1 WO 1999041407A1 EP 9900876 W EP9900876 W EP 9900876W WO 9941407 A1 WO9941407 A1 WO 9941407A1
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WO
WIPO (PCT)
Prior art keywords
amino acid
acid sequence
peptide
active ingredient
protein
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
PCT/EP1999/000876
Other languages
German (de)
English (en)
Inventor
Christof Antz
J. Peter Ruppersberg
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.)
Otogene AG
Original Assignee
Otogene 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.)
Filing date
Publication date
Application filed by Otogene AG filed Critical Otogene AG
Priority to EP99911661A priority Critical patent/EP1054997A1/fr
Priority to AU30274/99A priority patent/AU3027499A/en
Priority to JP2000531588A priority patent/JP2002503481A/ja
Publication of WO1999041407A1 publication Critical patent/WO1999041407A1/fr
Anticipated expiration legal-status Critical
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
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1055—Protein x Protein interaction, e.g. two hybrid selection
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/025—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B30/00—Methods of screening libraries
    • C40B30/04—Methods of screening libraries by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
    • G01N33/542—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with steric inhibition or signal modification, e.g. fluorescent quenching
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
    • G01N33/6845—Methods of identifying protein-protein interactions in protein mixtures

Definitions

  • the invention relates to a method for the development of an active substance which, preferably intracellularly, is able, directly or indirectly, to influence the function of physiologically active peptides or proteins.
  • the active substances developed should be applicable in particular with the aid of so-called gene therapy, and the function of the peptides / proteins should preferably be inhibited by this function.
  • the object of the invention is to avoid the disadvantages of the prior art.
  • the aim is to provide a process for developing an active substance defined above, in which an optimized amino acid sequence is obtained with comparatively less effort using systematic use of certain process steps.
  • This amino acid sequence or the active ingredient resulting from it should preferably be suitable for inhibiting physiologically active peptides / proteins.
  • the simplest possible application of the active ingredient with high effectiveness is to be achieved.
  • the amino acid sequence obtained by the method should act on the physiologically active target peptide / protein preferably at least as well as its physiological interaction partner.
  • the method mentioned at the outset is essentially characterized by two method steps.
  • a target peptide or protein that physiologically participates or can participate in a peptide-peptide or protein-protein interaction is identified and selected.
  • an amino acid sequence is derived or screened, which interacts with the target peptide / target protein or its section or sections with high affinity. This interaction is preferably at least as good, in particular better than that of the target peptide / target protein / section with its physiological interaction partner.
  • the derivation of the amino acid sequence preferably comprises the selection of at least one starting structure of an amino acid sequence, this selection preferably at the beginning of the
  • the starting structure can be, in particular, the amino acid sequence of a physiological interaction partner of the target peptide / target protein or one of its subsections or an amino acid sequence derived with the aid of molecular modeling. If the derivation of the amino acid sequence takes place in several process runs, the selection of one of the two start structures determined in the above-mentioned ways is particularly appropriate during the first process run. In these cases, the (partially optimized) amino acid sequences resulting from the previous process run can then be used as the starting structure for the further process runs.
  • the method according to the invention is further characterized in that a number of degenerate nucleic acid sequences are determined and selected on the basis of the starting structure (s). These code for the corresponding variations of the amino acid sequence selected as the starting structure.
  • This procedure provides an at least manageable number of sequences, the examination of which is practically possible with the aid of a screening method.
  • the starting structure it is achieved that the nucleic acid sequence does not have to be completely degenerate, but can at least be planned within limits. The variability mentioned at the outset can thus be significantly reduced. Normally, selecting the amino acid sequence of a known physiological interaction partner as starting structure with low variability will be sufficient as in the case of an amino acid sequence resulting from molecular modeling as starting structure.
  • the degenerate nucleic acid sequence obtained (possibly also several) is then preferably a known one Subject to screening methods with the aid of which the interaction between target peptide / target protein / partial section and the amino acid sequences which correspond to the individual degenerate nucleic acid sequence can be determined. Because of this screening step, a certain number will appear
  • the nucleic acid sequences that show the best interaction and have been selected accordingly i.e. in a conventional screening method, the corresponding yeast cells are usually sequenced after cloning. This sequencing then necessarily results in the associated amino acid sequences.
  • the sequences obtained in this way are preferably compared with one another and possibly also with existing sequences in order to obtain similarities or common structural motifs that can be useful for a possible further optimization of the amino acid sequence.
  • the associated amino acid sequences / peptides / proteins are then synthesized for the selected sequenced nucleic acid molecules by customary methods.
  • the peptides can also be expressed quantitatively in cells.
  • the exact interaction structure of the amino acid sequences obtained in this way is then determined using NMR spectroscopy. As already stated, this interaction structure is formed between the target peptide / target protein / partial section and the synthesized amino acid sequence. For better comparability of the results, one or more of the partial sections of the target peptide / target protein can also be produced synthetically for this or another process step.
  • the overall process can be carried out at least twice, preferably several times.
  • the amino acid sequence sought can be gradually optimized until either a sufficiently high affinity or even a better affinity than that of the physiological interaction partner is reached.
  • the user of the process is free to choose the process runs at a specific time, i.e. terminate at a certain degree of optimization of the amino acid sequence.
  • an amino acid sequence obtained (previously optimized or partially optimized) from earlier process steps with already good interaction / affinity is used in further process steps as a so-called petitor. This applies in particular to the implementation of the screening process at which then the further optimized sequences can be compared directly with the competitor.
  • the method according to the invention can be carried out in particular for the derivation of amino acid sequences for several subsections of a target peptide / target protein.
  • the overall affinity and specificity of the derived structure can thus be further improved, as will be explained in the following.
  • the required process steps for deriving the amino acid sequence per section are carried out with the abovementioned frequency of three to twenty times, preferably three to ten times.
  • the derived amino acid sequence usually comprises the smallest possible number of amino acids in order not to make the derivation of the amino acid sequence too complex.
  • a minimum number of amino acids will be required to ensure the required interaction with the target protein / target peptide / section. It is therefore preferred that the deduced amino acid sequence comprise less than 30 amino acids, preferably less than 15 amino acids.
  • Sub-sections of the target peptide / target protein can in principle be taken into account any amino acid sequences which are involved in a corresponding physiological protein-protein or peptide-peptide interaction. However, so-called helix or ⁇ -sheet structures are particularly suitable here. If, as in many cases, there are several active sections within a target peptide / protein, these can therefore preferably be mentioned Structures are selected and derived for these amino acid sequences.
  • the invention is based on the identification and selection of a target peptide / target protein or one or more of its subsections which are suitable for a physiological interaction with other peptides, proteins and the like. come into question.
  • the terms “peptide” and “protein” selected in the invention are intended to encompass in the broadest sense any amino acid sequence as known to the person skilled in the art.
  • an amino acid sequence is derived from this identified and selected amino acid sequence that is capable of an interaction, preferably an interaction that is at least as good as that between the sequence presented and its physiological interaction partner.
  • the starting point of the development is an already known peptide-peptide interaction, an interaction resulting from an earlier process or a purely computer-modeled structure.
  • the background of the invention is therefore a combination of a largely random screening and a deterministic modeling strategy. The combination of these two techniques makes it possible to choose from the vast number of possible sequences to select a series of, for example, 10 particularly good sequences.
  • vectors can be completely dispensed with. Accordingly, the half-life of a therapeutic effect is determined by the properties of the active ingredient and its target molecule and not by that of the gene therapy vehicle.
  • the required concentration of the active ingredient or the amino acid sequence which is usually in the nanomolar range, can already be achieved with a single copy of a corresponding DNA in a cell.
  • the invention further comprises an active substance, preferably an intracellular active substance, which is capable of influencing the function of physiologically active peptides or proteins directly or indirectly.
  • this active ingredient results from the method according to the invention already described or from one of its embodiments. There is a direct influence by the active ingredient, for example, if the amino acid sequence derived by the method according to the invention is used, ie this 10
  • the desired interaction occurs "directly".
  • An indirect influence is present, for example, when a nucleic acid molecule coding for the desired amino acid sequence is used as the active ingredient, which is introduced into the corresponding cell and which expresses the optimized amino acid sequence.
  • the active ingredient according to the invention is in particular an amino acid sequence obtainable by the method according to the invention or one of its embodiments.
  • This amino acid sequence is in particular a peptide which preferably has fewer than 15 amino acids.
  • the active substance can contain at least two (optimized) amino acid sequences. In principle, these can exist independently of one another and, for example, interact with corresponding subsections of a target peptide. However, it is preferred if the at least two amino acid sequences are linked to one another via physiologically largely inactive molecular intermediate members. This is in particular a covalent connection between the amino acid sequences and the pontics. A further increase in affinity is achieved by using such "covalent linkers".
  • the length of the molecular links between the amino acid sequences is the distance between the active sections in the target peptide or, under certain circumstances, also between different ones
  • Molecules can be constructed after structural analysis of individual receptor areas (partial section) or the entire receptor (target peptide). If, as in the present case, the derivation of amino acid sequences is used, poly-glycine or poly-alanine * chains of corresponding length can be inserted in a simple manner.
  • nucleic acid molecules preferably recombined nucleic acid molecules, which code for at least one amino acid sequence obtainable by the method according to the invention.
  • This can be a DNA molecule, cDNA molecule or RNA molecule. If molecular links are used as a linker between several amino acid sequences, the nucleic acid molecules additionally code for these linker chains.
  • the active substance according to the invention can be in the form of a so-called vector or vehicle which carries at least one of the nucleic acid molecules already described which code for the amino acid sequence.
  • vectors can be the usual viral and non-viral vectors as are known to the person skilled in the art. If a viral vector is used, a retro virus, an adenovirus or an adeno-associated virus can be used. In addition to non-viral vectors in which no viral DNA is involved, the packaging of the nucleic acid molecules in the so-called liposomes or lipoplexes should be emphasized.
  • non-viral vectors or liposomes / lipoplexes are preferred in order to avoid the known disadvantages of viral vectors, some of which are described at the beginning.
  • active ingredients can be made available with very high effectiveness, which in principle are viral without use 12
  • Vectors can be introduced into a cell.
  • the advantage of the invention is quite fundamentally that active ingredients, in particular amino acid sequences with nanomolar affinity, can be made available.
  • the invention comprises a pharmaceutical composition or a medicament that contains at least one of the described active ingredients, i.e. an optimized amino acid sequence containing the coding nucleic acid molecule or a vehicle carrying the nucleic acid molecule, and a pharmaceutically acceptable carrier.
  • Fig. 1 shows the protein structure of a complex of the cyclin-dependent kinase CDK2, the cyclin A and their
  • Fig. 2 shows the interaction between a section of cyclin A and a section of p27 Kl P 1 and
  • Fig. 3 shows the flow of certain embodiments of the method according to the invention in a schematic representation.
  • the complex is selected from the cyclin-dependent kinase CDK2, cyclin A and its inhibitor p27 Kl P 1 .
  • This structure is disclosed in the publication by AA Russo et al in Nature, 382, 325-331 (1996) and is shown in FIG. 1.
  • the aim of using the method according to the invention is to inhibit or prevent an interaction of the p27 ⁇ 1 P ⁇ - with the other two proteins.
  • an amino acid sequence is to be derived which preferably interacts better than the p27 Kl P 1 with the other two proteins, preferably the cyclin A.
  • Fig. 1 the places where peptide-peptide interaction between p27 Kl P 1 and CDK2 or Cyclin A takes place are highlighted in black.
  • the binding / interaction denoted by a is a binding site which was initially identified as a binding site with high affinity.
  • binding sites exist between an o ⁇ -helix of cyclin A and an o ⁇ -helix of p27 K; "-P 1 (binding / interaction b), a ⁇ -sheet structure of CDK2 and a ⁇ -turn of p27 Kl P 1 (binding / interaction c), a ⁇ -strand of CDK2 and a ⁇ -strand of p27 Kl P 1 (binding / interaction d)
  • another helix of p27 Kl P 1 occupies the so-called ATP binding site.
  • Fig. 2 shows the interaction between the two o ⁇ helices of cyclin A and p27 Kl P 1 , the amino acids 38 to 49 being involved on the p27 Kl P 1 side and theo ⁇ -5 helix on the cycline A side .
  • Fig. 2 shows that the fit between the two interacting structures is not perfect and therefore there will not be a very high affinity between the two interacting partners.
  • the method according to the invention is now intended to replace one of the two helices, in particular the helix of p27 Kl P 1, with an optimized peptide structure (amino acid sequence) which then binds the other helix, preferably that of cyclin A, with high affinity.
  • This optimized peptide structure is then expressed intracellularly as a peptide of, for example, and preferably up to 15 amino acid length, by an artificially introduced gene.
  • the concentration of the amino acid sequence in the cell should only be approximately the same as the concentration of p27 ⁇ i P 1 , which can be estimated at 10 nM / 1 or approximately 1,000 to 10,000 molecules per cell. As described, a single copy of a DNA encoding the amino acid sequence per cell could be sufficient to achieve such a concentration.
  • the starting structure is varied with the help of molecular modeling.
  • the (spatial) peptide structure of at least one, preferably many different, predetermined amino acid sequences is determined in a manner known per se and their fit with the corresponding subsection of the target peptide is examined. Based on molecular modeling 16
  • the screening process is the FRET yeast two hybrid process. From about 10 8 to 10 9 cells with clones of different sequences, a number of the best cells, preferably about 10, are selected in which the interaction between the peptides, measured by fluorescence resonance energy transfer (FRET), is strongest or is best. The cells with the best interaction are cloned and the corresponding nucleic acid sequences or the associated amino acid sequences are determined by sequencing. If one now compares, as also shown in FIG. 3, whether similarities between the selected sequences are recognizable or whether there are other starting points for common structural motifs, this information can be used for the further implementation of the method.
  • FRET fluorescence resonance energy transfer
  • the FRET yeast two hybrid method which can be used according to FIG. 3 is essentially based on the fact that the two peptides, the interaction of which is to be investigated with one another, are coupled to different fluorescent components.
  • fluorescence resonance energy transfer FRET
  • FACS fluorescence-activated cell sorting
  • BFP blue fluorescent protein
  • the final analysis of the protein structures and their interaction takes place with the aid of an NMR spectrometer.
  • other detection methods can also be used, such as the X-ray structure analysis of a crystallized peptide or protein complex. Due to the fact that the analysis of the peptide structure with the NMR spectrometer requires a comparatively high expenditure of time, the corresponding process step occupies an important position in the overall process.
  • the method according to the invention can be understood as a combination of a random screening and a deterministic modeling strategy. This procedure used here is based on the mathematical sequence of certain fitting algorithms, in which a random variation and a targeted change in the parameters also take place alternately 19

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  • Health & Medical Sciences (AREA)
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  • Proteomics, Peptides & Aminoacids (AREA)
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  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne un procédé de développement d'un principe actif qui, de préférence, peut influer directement ou indirectement sur la fonction de peptides ou de protéines physiologiquement actifs. Cette influence sur la fonction des peptides ou protéines doit se traduire, de préférence, par une inhibition de cette fonction. Selon ledit procédé, d'une part on identifie et on sélectionne un peptide cible ou une protéine cible qui participe ou peut participer physiologiquement à une interaction peptide-peptide ou protéine-protéine. Habituellement, il est nécessaire de ne procéder qu'à l'identification ou à la sélection d'un ou de plusieurs fragments de ce peptide ou de cette protéine. D'autre part, on dérive ou on crible une séquence nucléotidique qui interagit avec le peptide cible ou la protéine cible ou avec les fragments de ceux-ci, avec une affinité élevée. La séquence nucléotidique dérivée interagit, de préférence, au moins aussi bien et en particulier mieux avec le peptide cible qu'avec son partenaire physiologique d'interaction.
PCT/EP1999/000876 1998-02-11 1999-02-10 Procede de developpement d'un principe actif pharmaceutique Ceased WO1999041407A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
EP99911661A EP1054997A1 (fr) 1998-02-11 1999-02-10 Procede de developpement d'un principe actif pharmaceutique
AU30274/99A AU3027499A (en) 1998-02-11 1999-02-10 Method for developing a pharmaceutical active ingredient
JP2000531588A JP2002503481A (ja) 1998-02-11 1999-02-10 薬学活性成分の創製方法

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE1998105334 DE19805334A1 (de) 1998-02-11 1998-02-11 Verfahren zur Entwicklung eines pharmazeutischen Wirkstoffes
DE19805334.7 1998-02-11

Publications (1)

Publication Number Publication Date
WO1999041407A1 true WO1999041407A1 (fr) 1999-08-19

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Application Number Title Priority Date Filing Date
PCT/EP1999/000876 Ceased WO1999041407A1 (fr) 1998-02-11 1999-02-10 Procede de developpement d'un principe actif pharmaceutique

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EP (1) EP1054997A1 (fr)
JP (1) JP2002503481A (fr)
AU (1) AU3027499A (fr)
DE (1) DE19805334A1 (fr)
WO (1) WO1999041407A1 (fr)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991019818A1 (fr) * 1990-06-20 1991-12-26 Affymax Technologies N.V. Banque de peptides et systemes de triage
WO1996023899A1 (fr) * 1995-02-01 1996-08-08 University Of Massachusetts Medical Center Procedes de selection d'un peptide aleatoire se liant a une proteine cible
WO1997020078A1 (fr) * 1995-11-30 1997-06-05 Maxygen, Inc. Procede d'elaboration de polynucleotides presentant des caracteristiques desirees par selection iterative et recombinaison
WO1997027212A1 (fr) * 1996-01-23 1997-07-31 Rigel Pharmaceuticals, Inc. Methodes de criblage de peptides effecteurs intracellulaires transdominants et de molecules d'arn

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5077195A (en) * 1985-03-01 1991-12-31 Board Of Reagents, The University Of Texas System Polypeptides complementary to peptides or proteins having an amino acid sequence or nucleotide coding sequence at least partially known and methods of design therefor
DE69126020T2 (de) * 1990-02-14 1997-12-11 Receptor Lab Inc Methode zur herstellung und prüfung von verwendbaren peptiden
JPH07502111A (ja) * 1991-08-21 1995-03-02 レセプター　ラボラトリーズ　インコーポレイテッド 有用なペプチドの製造ならびにスクリーニング方法
DE4343527A1 (de) * 1993-12-16 1995-06-22 Schering Ag Verfahren zur Identifizierung von Stoffen mit potentieller herbizider oder wachstumsregulatorischer Wirkung mittels pflanzlicher Transporterproteine, Verwendung der Transporterproteine sowie Substanzen mit herbizider und wachstumsregulatorischer Wirkung
AU722840B2 (en) * 1996-03-30 2000-08-10 Medical Research Council Protein phosphatase-1 catalytic subunit interactions

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991019818A1 (fr) * 1990-06-20 1991-12-26 Affymax Technologies N.V. Banque de peptides et systemes de triage
WO1996023899A1 (fr) * 1995-02-01 1996-08-08 University Of Massachusetts Medical Center Procedes de selection d'un peptide aleatoire se liant a une proteine cible
WO1997020078A1 (fr) * 1995-11-30 1997-06-05 Maxygen, Inc. Procede d'elaboration de polynucleotides presentant des caracteristiques desirees par selection iterative et recombinaison
WO1997027212A1 (fr) * 1996-01-23 1997-07-31 Rigel Pharmaceuticals, Inc. Methodes de criblage de peptides effecteurs intracellulaires transdominants et de molecules d'arn

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AU3027499A (en) 1999-08-30
EP1054997A1 (fr) 2000-11-29
JP2002503481A (ja) 2002-02-05
DE19805334A1 (de) 1999-08-12

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