Classifications

• • 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
  • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
  • G01N33/56983—Viruses
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
  • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • A61K38/162—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from virus
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/12—Antivirals
  • A61P31/20—Antivirals for DNA viruses
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents

Definitions

• the present invention relates to a formulation comprising at least one papillomavirus-specific protein with about 0.3 to about 4 M of a salt at a pH of about 7.3 to about 7.45.
• the papilloma viruses also called wart viruses, are double-stranded DNA viruses with a genome size of about 8000 base pairs and an icosahedral capsid with a diameter of approx. 55 nm.
• human papilloma virus some of which, for example, HPV 16, HPV-18, HPV-31, HPV-33, HPV-39, HPV-45, HPV-52 or HPV-58, malignant tumors and others, e.g. HPV-6, HPV-1 1 or HPV-42, can cause benign tumors
• the genome of the papillomaviruses can be divided into three areas.
• the first area concerns a non-coding region that contains regulatory elements for the transcription and replication of the virus.
• the second region so-called E- (early) region, contains different protein-coding sections E1- E7, of which, for example, the E6 and E7 proteins are responsible for the transformation of epithelial cells and the E1 protein controls the DNA copy number.
• the E6 and E7 regions are so-called oncogenes, which are also malignant in malignant cells Cells are expressed.
• the third region also known as the L (late) region, contains two protein-coding sections L1 and L2 which code for structural components of the virus capsid.
• the Ll protein is present to over 90% in the viral capsid, whereby the Ratio of Ll L2 is generally 30 1
• HPV-6 and HPV-11 are blamed for genital warts
• some types of papilloma virus such as HPV-16, HPV-18, HPV-31, HPV-33, HPV-39, HPV-45, HPV-52 and HPV-58 are included malignant tumors of the anogenital tract
• HPV-16 is associated with cervical cancer in over 50% of cases (Cervical carcinoma)
• HPV-16 is the main risk factor for the formation of cervical neoplasms.
• the immune system also plays an important role in the progress of the disease.
• E7 protein is considered as a potential tumor antigen and as a target molecule for activated T cells.
• induced cellular immune response in the patient is apparently not strong enough to influence the course of the disease.
• the immune response may be enhanced by suitable vaccines
• VLPs virus-like particles
• the VLPs could be used to generate neutralizing antibodies in various animal systems
• virus-neutralizing antibodies are of less clinical importance if the virus infection has already taken place, since a virus-specific cytotoxic T-cell (CTL) response seems to be necessary for the inhibition of virus-infected cells.
• CTL cytotoxic T-cell
• CVLPs Papillomavirus-like particles
• Some CVLPs induce an E7-specific CTL response in mice, although experiments failed To induce antibodies against E7 by immunizing mice with CVLPs (Muller, M et al (1997), supra).
• neutralizing antibodies from HPV-associated To limit the immune diseases in patients to the administered L1 protein (Muller, M et al (1997), supra) CVLPs are, however, still interesting for the development of a vaccine, since the E7 presented via class I MHC molecules -Proteins from tumor cells were target molecules from CTLs
• VLPs or CVLPs are generally produced genetically by expression of the corresponding genes coding for one or more L proteins or L and E proteins in suitable expression systems.
• the corresponding genes are described, for example, by Kirnbauer, R et al (1994) J Virol 67 , 6929-6936 or obtainable from the EMBL database.
• the access numbers are, for example, for HPV18 PAPHPV18, for HPV31 PAPPPH31, for HPV33 PAPPPH33 or for HPV58 PAPPPH58.
• Suitable expression systems are, for example, genetically modified yeasts, for example Saccharomyces (cerevisiaiae), P (pastoris), Kluyvermyces (lactis), Schizosaccharomyces (pombe) or Hansenula (polymorpha) (Carter, JJ et al (1991), Virology, 182, 513), insect cells, such as, for example, Trichoplusia ni High Five (see, for example, Muller, M et al (1997), supra) or prokaryotic cells (see, for example, WO 96/1 1272).
• yeasts for example Saccharomyces (cerevisiaiae), P (pastoris), Kluyvermyces (lactis), Schizosaccharomyces (pombe) or Hansenula (polymorpha)
• insect cells such as, for example, Trichoplusia ni High Five (see, for example, Muller, M et al (1997), supra) or prokary
• inclusion bodies inclusion bodies
• capsomers further purification steps are necessary after expression
• capsids and capsomers A major problem with the use of capsids and capsomers as medicaments is their poor solubility.
• capsids or capsomers of HPV-16 tend to aggregate, as a result of which the solubility is substantially reduced.
• the partly low solubility of the capsids or capsomers not only leads to a loss of yield, but also to difficult use as a drug or diagnostic.
• a degradation of the C-terminus of the Ll protein can sometimes be observed, which leads to inhomogeneous leads material that is not suitable for approval as a drug or diagnostic
• WO 98/44944 describes an HPV antigen formulation containing a vaccine component, a salt at physiologically acceptable concentrations and a non-ionic detergent at physiologically acceptable concentrations
• the present invention therefore relates to a formulation comprising at least one late protein (L protein) of one or more papillomaviruses and / or at least one early protein (E protein) of one or more papillomaviruses and about 0.3 to about 4 M , preferably about 0.4 to about 2.5-3 M, in particular about 0.4-0.5 to about 1-2 M, especially about 1 to about 2 M of a salt at a pH of about 7, 3 to about 7.45, preferably about 7.4, and, if appropriate, suitable additives and / or auxiliaries, preferably without any additives and / or auxiliaries
• the term “formulation” is understood to mean a composition in the form of a solution or a suspension of the papillomavirus-specific proteins mentioned, with immunoreactive papilla do not sediment mavirus-specific proteins in general and in particular up to a maximum of about 5000 g
• the salt is generally an alkali or alkaline earth metal salt, preferably a halide or phosphate, in particular an alkali halide, especially NaCl and / or KC1.
• a halide or phosphate in particular an alkali halide, especially NaCl and / or KC1.
• the use of NaCl is particularly preferred for the preparation of a pharmaceutical formulation
• the pH of the composition is generally adjusted with a suitable organic or inorganic buffer, such as preferably with a phosphate buffer, Tris buffer (Tris (hydroxymethyl) aminomethane), HEPES buffer ([4- (2 -Hydroxyethyl) -piperazino] -ethanesulfonic acid) or MOPS buffer (3-morpholino-l-propanesulfonic acid).
• a suitable organic or inorganic buffer such as preferably with a phosphate buffer, Tris buffer (Tris (hydroxymethyl) aminomethane), HEPES buffer ([4- (2 -Hydroxyethyl) -piperazino] -ethanesulfonic acid) or MOPS buffer (3-morpholino-l-propanesulfonic acid).
• a suitable organic or inorganic buffer such as preferably with a phosphate buffer, Tris buffer (Tris (hydroxymethyl) aminomethane), HEPES buffer ([4- (2 -Hyd
• composition according to the invention as a medicament or diagnostic agent, it is particularly preferred if the papillomavirus-specific proteins do not contain papillomavirus-unspecific epitopes, since this can reduce or prevent a papillomavirus-unspecific immune response
• L-protein and E-protein mean both the Vol GmbH proteins and their mutants, such as, for example, deletion mutants
• the composition according to the invention contains a deleted L protein, preferably a deleted L1 and / or L2 protein.
• the deletion has the advantage that other proteins, for example PapiUomavirus-specific E protein sequences, can be inserted into the deleted area , whereby the field of application of the composition according to the invention can be expanded.
• the C-terminal deletion has the advantage that the efficiency of the The formation of virus-like particles can be increased since the nuclear localization signal located at the C-terminus is deleted.
• the C-terminal deletion is therefore preferably up to approx. 35 amino acids, in particular approx. 25 to approx.
• a 32 amino acid C-terminal deletion of the HPV-16 L1 protein and an approximately 26 amino acid long C-terminal deletion of the BPV-1 Ll protein are sufficient to form virus-like particles to be able to increase this at least about ten times
• the E protein is also deleted, in particular the E6 and / or E7 protein.
• the C-terminal part of the E protein is deleted, preferably the C-terminal part of the E7 Proteins, since these constructs can preferably form capsomers and / or capsids in conjunction with deleted L-protein.
• a particularly preferred construct is, for example, E7 with the N-terminal amino acids 1 to about 60, since this construct contains a mouse epitope for the activation of cytotoxic T lymphocytes, which is located in the region of amino acids 49-57.
• Another preferred construct is E7 with the N-terminal amino acids 1 to about 55, which forms capsomers and capsids in conjunction with deleted L-protein, since this construct presumably does not contain E7-specific sequences in the region of amino acids 56-70, which form the formation of capsids
• a Ll protein of HPV-16 deleted by 32 amino acids C-terminal, which is linked to an E7 protein of HPV-16 with amino acids 1-55 or 1-60 is particularly preferred.
• a particularly preferred embodiment of the present invention is therefore a Ll ⁇ E7 ⁇ -x fusion protein, preferably in the form of a CVLP, in particular of HPV16, where x is an integer from 55 up to and including 60, and in particular an L1 ⁇ CE7] .55 or Ll ⁇ CE7 ⁇ .6o fusion protein
• the L protein to the E protein, for example in the form of a fusion protein.
• the described papillomavirus-specific proteins are in the form of a capsid and / or Capsomers are present, since the immune response can be significantly increased by the capsids and / or capsomers and in particular by the proportion of L protein.
• Fusion proteins which are suitable for capsid and / or capsomer formation are therefore, for example, fusion proteins from deleted L1 and E7, E6 and / or El preferred
• capsids are viral or virus-like structures in a generally icosahedral form, which are generally composed of approximately 72 capsomers
• capsomers are assembled proteins containing at least one papillomavirus structural protein, preferably L1 or deletions of L1.
• L1 or deletions of L1 preferably L1 or deletions of L1.
• 5 fusion proteins can assemble to form a capsomer, which in turn can assemble to form a capsid
• Proteins or peptides of human papillomavirus (HPV) and preferably of HPV-6, HPV-11, HPV-16, HPV-18, HPV-31, HPV-33, HPV are for the described constructs for the production of a human medicament or diagnostic agent -35, HPV-39, HPV-45, HPV-52 and / or HPV-58, in particular HPV-16, HPV-18, HPV-31 and / or HPV-45 suitable.
• the expression vectors can be, for example, prokaryotic or eukaryotic expression vectors.
• prokaryotic expression vectors for expression in E. coli for example, the vectors pGEM or pUC-Derviate (see, for example, WO96 / 11272).
• Examples of eukaryotic expression vectors for expression in Saccharomyces cerevisiae for example, the vectors p426Met25 or p426GALl (Mumberg et al (1994) Nucl Acids Res, 22, 5767-5768, Carter, JJ et al (1991) supra) and for expression in insect cells, for example baculovirus vectors, in particular the Autographa Californica virus, as in EP-B1-0 127 839 or EP- B1-0 549 721 (see, for example, also WO94 / 20137), and for expression in sucker cells, for example the vectors Rc / CMV and Rc / RSV or SV40 vectors, all of which are generally available.
• Baculovirus expression systems such as the Baculo Gold TM Transfection Kit from Pharmingen or the Bac-to-Bac TM Baculovirus Expression Systems from Gibco BRL.
• Other suitable expression systems are recombinant vaccinia viruses (see, for example, WO 93/02184)
• the expression vectors also contain promoters suitable for the respective host cell, such as, for example, the trp promoter for expression in E. coli (see, for example, EP -B 1-0 154 133), the ADH2 promoter for expression in yeasts ( Rüssel et al (1983), J Biol Chem 258, 2674-2682), the baculovirus polyhedrin promoter for expression in insect cells (see, for example, EP-B1-0 127 839 or US 5,004,687) or the early SV40 promoter or LTR - Promoters e.g. from MMTV (mouse mammary tumor virus, Lee et al (1981) Nature 214, 228-232)
• promoters suitable for the respective host cell such as, for example, the trp promoter for expression in E. coli (see, for example, EP -B 1-0 154 133), the ADH2 promoter for expression in yeasts ( Rüssel et al (1983), J Biol Chem 258, 2674-
• Suitable host cells are, for example, the E coli strains DH5, HB101 or BL21, the yeast strains Saccharomyces cerevisia, Pichia, Kluyvermyces, Schizosaccharomyces or Hansenula (Carter, JJ et al (1991), Virology, 182, 513), the insect cell line Lepidopteran, e.g. from Spodoptera frugiperda, Trichoplusia ni, Rachiplusia ou or Galleria Mellonela or the animal cells COS, C127, Vero, 293 and HeLa, all of which are generally available (see e.g. WO94 / 00152)
• the coding nucleic acids for the individual papillomavirus-specific proteins can, for example, be isolated and cloned from a gene bank via PCR (“polymerase chain reaction”) amplification.
• PCR polymerase chain reaction
• the genome of BPV-1 is under the GenBank Accession No X02346 or HPV-16 generally available under GenBank Accession No K02718.
• Amino acid long HPV 16 E7 protein is described, for example, by Seedorf et al.
• Virology 145, 181-185.
• Another method of obtaining the desired nucleic acids is to isolate the papillomavirus-specific genes directly from warts or tumors by means of PCR. Suitable primers for the E6 and E7 genes HPV-16 and HPV-18 are disclosed, for example, in WO93 / 21958. Further references for the desired nucleic acids are, for example, Kirnbauer, R et al (1994), supra and the clones mentioned above in the EMBL database
• the expression vector is constructed in such a way that the expressed fusion protein is not extended by any further amino acids caused by the vector. This is achieved, for example, by generating unwanted nucleotides by mutagenesis in a PCR reaction using suitable primer oligonucleotides code for additional amino acids, are removed (Ho et a (1989) Gene, 77, 51-59). In this way, a fusion protein is obtained which is free of additional amino acids and thus free of any additional foreign epitopes which can cause immunological side reactions
• Suitable additives and / or auxiliaries which serve, for example, to further stabilize the papillomavirus-specific protein in the composition according to the invention, are, for example, detergents, such as, for example, Triton-X-100 or sodium deoxycholate, but also polyols, such as, for example, polyethylene glycol or Glycerin, sugar, such as B sucrose or glucose, zwitterionic compounds, such as. B amino acids such as glycine or in particular taurine or betaine and / or protein, such as bovine or human serum albumin. Detergents, polyols and / or zwitterionic compounds are preferred
• protease inhibitors such as aprotinin, ⁇ -aminocaproic acid or pepstatin A
• Another object of the present invention is a process for the preparation of the formulation according to the invention, the PapiUomavirus-specific protein described above, for example in solution containing about 0.3 to about 4 M, preferably about 0.4 to about 2.5 to 3 M, in particular approx. 0.4 - 0.5 to approx. 1 - 2 M, especially approx. 1 to approx. 2 M of a salt with a corresponding pH value of approx. 7.3 to approx. 7.45, preferably approx. 7.4 as well as suitable ones
• Additives and auxiliary substances are introduced and / or dialyzed against the composition described.
• the formulation can preferably be stored in a stable manner at about 4 ° C. or above all at about -80 ° C. over a longer period, for example 1-2 months or longer
• the formulation according to the invention is suitable as a medicament or diagnostic agent.
• the present invention therefore also relates to the use of the formulation according to the invention as a medicament or diagnostic agent.
• the formulation according to the invention is preferably adjusted to a concentration of approximately 0.45 M.
• the medicament does not contain an adjuvant, ie no substance which enhances the immunogenicity of the papillomavirus-specific protein, since in particular in the presence of an L protein, especially of Ll, the immunogenicity is already sufficiently enhanced. This property is particularly important in the approval as a drug or diagnostic, because the only immunostimulating materials approved by the regulatory authorities are currently aluminum salts
• the medicinal product is particularly suitable for avoiding and / or treating papillomavirus-specific benign or malignant tumors, in particular malignant tumors such as, for example, laryngeal, cervical, penile, vulvar or anal carcinoma, and the diagnostic agent for diagnosing one or several pairs pillomavirus infection (s)
• a diagnostic agent is immunodiagnostics known to the person skilled in the art, for example an ELISA for measuring papillomavirus-specific antibodies (see, for example, Voller, A et al (1976) Bull World Health Organ, 53, 55-63) or a skin test according to e.g. Hopfl et al (1991) Lancet 1, 373-374)
• the medicament can be administered orally, parenterally, for example subcutaneously, intramuscularly or via the mucous membrane, in liquid or suspended form, in the form of an elixir or as capsules, preferably as an injection or infusion solution an adjuvant can be dispensed with, which is particularly advantageous
• Another object of the present invention therefore relates to the use of the formulation according to the invention as an injection or infusion solution
• Injection solutions are generally used when only relatively small amounts of a solution or suspension, for example about 1 to about 20 ml, are to be supplied to the organism.
• Infusion solutions are generally used when a larger amount of a solution or suspension, for example a or several liters, should be administered Since, in contrast to the infusion solution, only a few milliliters are administered with injection solutions, slight deviations from the pH value and from the osmotic pressure of the blood or the tissue fluid during the injection do not, or only insignificantly with regard to the Sensation of pain noticeable. Dilution of the formulation according to the invention before use is therefore generally not necessary.
• the formulation according to the invention should be diluted shortly before application so that an isotonic solution can be obtained
• the ionic solution is a 0.9% sodium chloride solution.
• the dilution can be done, for example, with sterile water during the application, for example via a so-called bypass
• the main advantage of the present invention is that the formulation according to the invention essentially does not lead to the precipitation of immunoreactive papillomavirus-specific protein. In particular, more than about 90%, especially more than about 95% of the protein remain in solution and do not fall for one Period of at least approx. 12 hours from.
• the immunoreactive PapiUomavirus-specific protein cannot be sedimented significantly by centrifugation at a maximum of 5000 g.
• the formulation remains homogeneous and stable over a longer period of approx. 1-2 months and longer
• HPV 16L1 ⁇ C * E7 1-55 was prepared according to Muller, M et al (1997), supra.
• the HPV-16L1 open reading frame (ORF) was derived from the plasmid HPV-16-114 / k-Ll / L2-pSynxtVr ( Kirnbauer, R et al (1994) J Virol 67, 6929) with the restriction endonuclease BglII and cloned into the vector pUC19 (New England Biolabs) in the BamHI site.
• Two primers were constructed for the production of HPV-16L1 ⁇ C. 16L1 ORF are complementary.
• the first primer has the sequence AAAGATATCTTGTAGTAAAAATTTGCGTCCTAAAGGAAAC and the second primer AAAGATATCTAATCTACCTCTACAACTGCTAAACGCAAAAAACG
• Both primers code 5 'an EcoRV restriction enzyme interface.
• the EcoRV site is followed by a TAA translation stop codon in order to delete the last 34 amino acids of the HPV16L1 ORF.
• the PCR reaction was carried out to remove the entire L1 ORF and the whole Vector to amplify.
• the linear product was digested with EcoRV, circularized with T4 DNA ligase and E coli DH5 ⁇ cells transformed. The clones were analyzed for the presence of an EcoRV site.
• the resulting construct pUCHPV16Ll ⁇ C was used to isolate the ORF of HPV16E7 1-50 in to clone the EcoRV site
• Primers with a 5 PrimcoRV restriction enzyme interface were used to clone the fragment.
• the following primer pair was used: AAAAGATATCATGCATGGAGATACACCTACATTGC and
• the PCR products were cleaved with EcoRV and inserted into the EcoRV site of the modified Ll gene
• Spodoptera frugiperda (Sf9) cells were grown as monolayers or in suspension culture in TNM-FH insect medium (Sigma, Deisenhofen) with 10% fetal calf serum and 2 mM glutamine.
• Recombinant baculoviruses HPV16L1 ⁇ CE7 1-55 were obtained by cotransfection of 10 ⁇ g and the recombinant 2 ⁇ g of linearized Baculo-Gold DNA (Pharmingen, San Diego, CA) transfected in Sf9 cells.
• Recombinant viruses were purified according to the manufacturer's instructions.
• 10 6 Sf9 cells were mixed with recombinant baculovirus and a moi (“multiplicity of infection ”) from 5 to 10 infected.
• Trichoplusia ni (TN) high five cells were grown at 27 ° C. to a density of 1-1.5 ⁇ 10 6 cells per ml in Ex-Cell 405 serum-free medium (JRH, Biosciences, Lennexa, KS)
• JRH, Biosciences, Lennexa, KS serum-free medium
• a 400 ml culture was harvested and infected with recombinant baculoviruses for one hour with periodic inversions with a moi of 2 to 5.
• Up to 240 ml of medium were added and the cells grew for 3 to 4 days.
• the cells were then pelleted and in 10 ml extraction buffer resuspended (25 mM Tris / HCl, pH 7.5, 500 mM NaCl, 1 mM EDTA) and sonicated for 45 seconds at 60 watts. After centrifugation at 10,000 rpm in the Sorvall SS34 rotor, the pellet was placed in the 6 ml extraction buffer dissolved, sonicated at 60 watts for 30 seconds and centrifuged again.
• 10 ml extraction buffer resuspended 25 mM Tris / HCl, pH 7.5, 500 mM NaCl, 1 mM EDTA
• the supernatants were combined and applied to a two-stage gradient composed of 40% (w / v) sucrose and 57.5% (w / v) CsCl after centrifugation in a SW-28 rotor at 27,000 rpm for two For one hour, the interphase and the CsCl layer were collected, adjusted to a CsCl density of 1.38 g / ml and centrifuged at 45,000 rpm for 16 hours.
• the gradients were fractionated and each fraction was subjected to Western blot with anti-HPVl ⁇ LlmAb Camvirl (Pharmingen, San Diego, CA)
• the reactive fractions were pooled and ultrafiltered with Centricon 30 microconcentrator (Amicon Corp Beverly, MA) against Hepes buffer (1mM Hepes, 149mM NaCl, 0.5mM KCl, pH 7.2) dialyzed and the presence of CVLPs confirmed by means of transmission electron microscopy.
• the concentration of LlE7 protein was approximately determined in an SDS gel which was stained with Coomassie blue by comparison with BSA standards
• a fraction containing virus-like particles isolated from high five cells by sucrose cushion and casium chloride equilibrium ultracentrifuge was used as a sample.
• the total protein concentration was 0.29 mg / ml and the CVLP concentration was 0.17 mg / ml

Landscapes

• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Virology (AREA)
• Chemical & Material Sciences (AREA)
• Engineering & Computer Science (AREA)
• Medicinal Chemistry (AREA)
• General Health & Medical Sciences (AREA)
• Immunology (AREA)
• Molecular Biology (AREA)
• Pharmacology & Pharmacy (AREA)
• Veterinary Medicine (AREA)
• Public Health (AREA)
• Animal Behavior & Ethology (AREA)
• Hematology (AREA)
• General Chemical & Material Sciences (AREA)
• Urology & Nephrology (AREA)
• Biotechnology (AREA)
• Biomedical Technology (AREA)
• Organic Chemistry (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Cell Biology (AREA)
• Food Science & Technology (AREA)
• Communicable Diseases (AREA)
• Tropical Medicine & Parasitology (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Epidemiology (AREA)
• Gastroenterology & Hepatology (AREA)
• Microbiology (AREA)
• Proteomics, Peptides & Aminoacids (AREA)
• Oncology (AREA)
• Physics & Mathematics (AREA)
• Analytical Chemistry (AREA)
• Biochemistry (AREA)
• General Physics & Mathematics (AREA)
• Pathology (AREA)
• Peptides Or Proteins (AREA)
• Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
• Micro-Organisms Or Cultivation Processes Thereof (AREA)

Applications Claiming Priority (3)

Publications (1)

Family

ID=7862147

Family Applications (1)

Country Status (8)

Families Citing this family (4)

Family Cites Families (4)

• 1998
  • 1998-03-24 DE DE19812940A patent/DE19812940A1/de not_active Withdrawn
• 1999
  • 1999-03-24 WO PCT/EP1999/001999 patent/WO1999048917A2/de not_active Application Discontinuation
  • 1999-03-24 AU AU35989/99A patent/AU3598999A/en not_active Abandoned
  • 1999-03-24 EP EP99917850A patent/EP1066321A2/de not_active Withdrawn
  • 1999-03-24 CA CA002323526A patent/CA2323526A1/en not_active Abandoned
  • 1999-03-24 AR ARP990101311A patent/AR014969A1/es unknown
  • 1999-03-24 JP JP2000537899A patent/JP2002507625A/ja active Pending
  • 1999-03-24 MX MXPA00009283A patent/MXPA00009283A/es unknown

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events