Antibodies specific to tumour necrosis factor, and application thereof

FIELD: pharmacology.

SUBSTANCE: invention concerns immunology and biotechnology. There is offered human monoclonal antibody specific to TNF-alpha containing light and heavy chain with appropriate CDR3 sites. There are described versions thereof including those based on heavy and light chains and coded by human genes VH3-33 and A30VK1 or VH3-53 and L2VK3 respectively. There are disclosed: the method for estimating the TNF-alpha content in the patient's sample with using specified antibodies, and application of antibodies for preparing a medical product. There are described: compositions for diagnostics and treatment of the conditions associated with TNF-alpha activity on the basis of antibodies. There is disclosed coding nucleic acid, a cell for making said antibodies and the method for making said antibodies.

EFFECT: application of the invention ensured high-affinity neutralizing monoclonal antibodies with improved Kd and IC50 in comparison with Infliximab, Adalimumab or Etanercept that can find application in medicine for treatment and diagnostics of the diseases associated with TNF-alpha hyperactivity.

35 cl, 13 dwg, 36 tbl, 14 ex

 

The text descriptions are given in facsimile form.

1. Monoclonal antibody of the person, which specifies the way associated with tumor necrosis factor alpha (TNF-alpha) and contains:
variable region of the heavy chain, which contains hypervariable section 3 (CDR3) of the heavy chain that has the amino acid sequence Glu Val Glu Ser Ala Met Gly Gly Phe Tyr Tyr Asn Gly Met Asp Val, and
variable region light chain containing hypervariable section 3 (CDR3) of the light chain that has the amino acid sequence Leu Gin His Lys Ser Tyr Pro Leu Thr.

2. Monoclonal human antibody according to claim 1, containing hypervariable section 2 (CDR2) of the heavy chain that has the amino acid sequence Val Ile Trp Ser Asp Gly Ser Ile Lys Tyr Tyr Ala Asp Ser Val Lys Gly.

3. Monoclonal human antibody according to claim 1 or 2, containing the hypervariable region 1 (CDR1) of the heavy chain that has the amino acid sequence of Ser Tyr Asp Met His.

4. Monoclonal human antibody according to claim 1 or 2, the amino acid sequence of the heavy chain which contains the amino acid sequence of SEQ ID NO: 70.

5. Monoclonal human antibody according to claim 1 or 2, the amino acid sequence of the heavy chain which contains the amino acid sequence of SEQ ID NO: 74.

6. Monoclonal human antibody according to claim 1 or 2, containing the hypervariable region 1 (CDR1) light chain that has the amino acid sequence Arg Ala Ser Gln Gly lle Arg Ile Asp Leu Gly.

7. Monoclonal human antibody according to claim 1 or 2, containing hypervariable section 2 (CDR2) light chain that is has the amino acid sequence Ala Ala Ser Thr Leu Gln Ser.

8. Monoclonal human antibody according to claim 1 or 2, the amino acid sequence of the light chain of which contains the amino acid sequence of SEQ ID NO: 72.

9. Monoclonal human antibody that specifically binds tumor necrosis factor alpha and contains:
heavy chain, which contains the amino acid sequence encoded by the gene VH3-33, or a conservative variant, and
light chain containing the amino acid sequence encoded by the gene light chain A30VK1 person, or a conservative variant.

10. Monoclonal human antibody that specifically binds to tumor necrosis factor alpha and contains:
variable region of the heavy chain, which contains hypervariable section 3 (CDR3) of the heavy chain that has the amino acid sequence Gly Glu Gly Gly Phe Asp Tyr, and
variable region light chain, which contains hypervariable section 3 (CDR3) of the light chain that has the amino acid sequence Gln Gln Tyr Asn Tyr Trp Trp Thr.

11. Monoclonal human antibody of claim 10, containing hypervariable section 2 (CDR2) of the heavy chain that has the amino acid sequence Val Ile Tyr Ser Gly Asp Arg Thr Tyr Tyr Ala Asp Ser Val Lys Gly.

12. Monoclonal human antibody of claim 10 or 11, containing the hypervariable region 1 (CDR1) of the heavy chain, which has sledovatelnot amino acids Arg Asn Tyr Met Ser.

13. Monoclonal human antibody of claim 10 or 11, the amino acid sequence of the heavy chain which contains the amino acid sequence of SEQ ID NO: 50.

14. Monoclonal human antibody of claim 10 or 11, containing the hypervariable region 1 (CDR1) light chain that has the amino acid sequence Arg Ala Ser Gln Ser Val Ser Ser Asn Leu Ala.

15. Monoclonal human antibody of claim 10 or 11, containing hypervariable section 2 (CDR2) light chain that has the amino acid sequence Gly Ala Ser Ile Arg Ala Thr.

16. Monoclonal human antibody of claim 10 or 11, containing hypervariable section 3 (CDR3) of the light chain that has the amino acid sequence Gln Gln Tyr Asn Tyr Trp Trp Thr.

17. Monoclonal human antibody of claim 10 or 11, the amino acid sequence of the light chain of which contains the amino acid sequence of SEQ ID NO: 52.

18. Monoclonal human antibody that specifically binds tumor necrosis factor alpha and contains:
heavy chain, which contains the amino acid sequence encoded by the gene VH3-53, or a conservative variant of the sequence and a light chain, which contains the amino acid sequence encoded by the gene light chain L2VK3 person, or a conservative variant of the sequence.

19. Monoclonal antibodies of man according to any one of claims 1, 2, 9-11 and 18, which connects Fnoa with a Kd of less than 10-10M

20. Monoclonal human antibody according to claim 19, which connects Fnoa with a Kd of less than 10-11M

21. Monoclonal human antibody according to any one of claims 1, 2, 9-11, 18, which is produced by line hybridoma.

22. Monoclonal human antibody according to any one of claims 1, 2, 9-11, 18, which produced SNO cells.

23. Monoclonal human antibody according to any one of claims 1, 2, 9-11, 18, which is an IgG1.

24. Monoclonal human antibody according to any one of claims 1, 2, 9-11, 18, which represents an IgG2.

25. How to evaluate the content of tumor necrosis factor alpha (α) in the sample taken from the patient, according to which the antibody against α according to any one of claims 1 to 24 is brought into contact with the biological sample taken from the patient, and determine the degree of binding of the indicated antibodies with α in the specified sample, and the concentration α in the specified sample is determined on the basis of the number α associated with the indicated antibody.

26. The method according A.25, where the biological sample is a blood.

27. The composition for the diagnosis of conditions associated with the activity and/or increased production of α containing an effective amount of the antibody according to any one of claims 1 to 24 or a functional fragment and a pharmaceutically acceptable nose is tel.

28. The use of antibodies according to any one of claims 1 to 24 for the preparation of a drug for the effective treatment of neoplastic diseases, immune-mediated inflammatory diseases or apoptosis induced by tumor necrosis factor in the animal.

29. Use p, where the specified neoplastic disease selected from the following group: breast cancer, ovarian cancer, gallbladder cancer, lung cancer, glioblastoma, stomach cancer, endometrial cancer, kidney cancer, colon cancer, pancreatic cancer, prostate cancer.

30. Use p, where the aforementioned immune-mediated inflammatory disease selected from the following group: rheumatoid arthritis, glomerulonephritis, atherosclerosis, psoriasis, restenosis, autoimmune disease, Crohn's disease, transplant rejection, septic shock, cachexia, anorexia, ankylosis spondylitis, multiple sclerosis.

31. Use p, where the aforementioned immune-mediated inflammatory disease selected from the group comprising rheumatoid arthritis, psoriasis and Crohn's disease.

32. Nucleic acid that encodes the antibody according to any one of claims 1 to 24.

33. A host cell for production of the antibody described in any one of claims 1 to 24, which contains a nucleic acid according p.

34. A method of producing antibodies, which includes
cultivar is the study of cells on p in terms when the specified nucleic acid is expressed with the formation of antibodies and isolating the antibody.

35. Composition for treatment of conditions associated with the activity and/or increased production of α containing an effective amount of the antibody according to any one of claims 1 to 24 or a functional fragment and a pharmaceutically acceptable carrier.



 

Same patents:

FIELD: medicine.

SUBSTANCE: invention is related to nucleic acids and multidomain proteins, which are able to bind vessel endotheliocyte growth factor (VEGF), and may be used in medicine. Recombinant method is used to produce polypeptide, which consists of component (R1R2)X and, unnecessarily, multidomain component (MC), which represents aminoacid sequence with length from 1 to 200 of amino acids, having at least one remainder of cysteine, where X≥1, R1 means antibody-like (Ig) domain 2 of VEGF receptor Llt-1, and R2 means Ig-domain 3 of VEGF receptor Flk-1. Produced fused polypeptide does not contain multidomain component in case, when X=2, and in case when X=1, multidomain component represents aminoacid sequence with length from 1 to 15 amino acids. Produced polypeptide is used in composition of pharmaceutical compound for VEGF-mediated disease or condition.

EFFECT: invention makes it possible to produce highly efficient trap of VEGF, special structure of which is suitable for local introduction into specific organs, tissues or cells.

16 cl, 3 tbl, 7 ex

FIELD: food industry.

SUBSTANCE: strain Streptococcus thermophilus which produces lactic acid is described. Sequence of nucleic acids made of the strain producing polysaccharides are also described as well as food or pharmaceutical composition and milk product containing such strain.

EFFECT: strain has strong structural properties.

16 cl, 4 dwg, 6 tbl, 5 ex

FIELD: medicine.

SUBSTANCE: invention relates to field of genetic engineering and medicine. Described is animal, non-human, having sequence of nucleic acid encoding presenilin 1, carrying mutations, corresponding to M233T and L235P mutations in PS1 protein of mouse. Animal also contains sequence of nucleic acid, encoding whole gene or part of gene, encoding APP. APP protein represents APP751, originates from human and carries mutations Swedish and London. Animal is intended for application in fight against Alzheimer's disease. Also described are PS1 protein and encoding it nucleic acid.

EFFECT: invention can be used in medicine for discovering compounds intended for Alzheimer's disease treatment.

20 cl, 50 dwg, 1 tbl, 8 ex

FIELD: chemistry; biochemistry.

SUBSTANCE: invention relates to biotechnology, specifically to production of versions of the Gla domain of human factor VII or human factor VIIa, and can be used in medicine. Amino acid sequence of the FVII or FVIIa version is obtained, which differs on 1 to 15 amino acid residues with amino acid sequence of the human factor VII (hFVII) or human factor VIIa (hFVIIa), in which a negatively charged amino acid residue is introduced by substitution in position 36. Obtained variants of FVII or FVIIa are used in a composition for treating mammals with diseases or disorders, where blood clotting is desirable.

EFFECT: invention allows for producing versions of FVII or FVIIa with high clotting activity and/or high activation of factor X, compared to natural form of hFVIIa.

42 cl, 3 dwg, 5 tbl, 11 ex

FIELD: medicine.

SUBSTANCE: vitamin K dependent protein is made by separating a cultivated eukaryotic cell that contains an expressing vector that contains a nucleic acid molecule coding vitamin K dependent protein and associated sequences regulating expression. The associated sequences contain the first promoter and the nucleic acid molecule coding gamma-glutamylcarboxylase, and the second promoter. The first promoter represents a pre-early promoter of human cytomegalovirus (hCMV), and the second promoter is a pre-early promoter SV40. Herewith the expressing relation of vitamin K dependent protein and gamma-glutamylcarboxylase is 10:1 to 250:1.

EFFECT: invention allows for making gamma-carboxylated vitamin K dependent protein in production quantities.

29 cl, 5 dwg, 6 tbl, 7 ex

FIELD: medicine.

SUBSTANCE: invention claims synthetic DNS molecule encoding L1 HPV58 protein, where codons are optimised for high expression level in yeast cell. Also invention claims expression vector, yeast host cell, HPV58 virus-like particle and method of its obtainment, and pharmaceutical composition including such VLP.

EFFECT: invention can be applied in medicine for efficient immune prevention of papillomavirus infection by neutralising antibodies and cell-mediated immunity, and for treatment of developed HPV infections.

10 cl, 10 dwg, 8 ex

FIELD: medicine.

SUBSTANCE: invention includes obtaining and applying the versions of allergens of 5 Pooideae group, which are characterised by decreased IgE reactivity in comparison to known wild-type allergens and at the same time by essentially retained reactivity in comparison to T-lymphocytes. Versions of allergens, as per the invention, do not have at least one section or a combination of sections corresponding to amino-acid sequence of sections Phl p5a 94-113 or 175-198, which is given in the description. Versions of allergens have been obtained by gene-engineering methods. In the invention is opened DNA molecule encoding the version of allergen, recombinant expression vector, host organism, expression version of allergen and method of obtaining the version of allergen by using the above described means. Such hypoallergic versions of allergens can be used as a remedy against allergies determined by allergens of 5 Pooideae group for specific immunotherapy (hyposensitisation) of the patients having grass pollen allergy or for preventive immunotherapy of grass pollen allergies by using pharmaceutical composition.

EFFECT: preparations based on versions of allergens, as per the invention, have decreased IgE reactivity and the retained reactivity in relation to T-lymphocytes.

12 cl, 17 dwg, 3 tbl

FIELD: chemistry.

SUBSTANCE: proposed is a recombinant single-strand trispecific antibody for treating tumours which express CEA. The said antibody consists of a series of three antibody fragments: anti-CEA-scFv, anti-CD3-scFv and VH CD28-antibody, linked by two intermediate linkers (intermediate linker Fc and intermediate linker HSA). If necessary, a c-myc-mark or (His)6-mark can be added at the C-end. Described is DNA, which codes the antibody, expression vector based on it and E.coli cell, containing the vector.

EFFECT: use of the invention is more beneficial in clinical use compared to bispecific antibodies and known trispecific antibodies, makes easier clearing and expression of an antibody, which can further be used in treating CEA-mediated tumours.

10 cl, 21 dwg, 11 ex

FIELD: chemistry; medicine.

SUBSTANCE: claimed are polypeptide and respective polynucleotide zcytor17lig and molecules of antibody against human zcytor17. Human zcytor17lig is novel cytokine. Claimed invention also relates to methods of protein obtaining, its application for stimulation of immune reaction in mammal. Described is method of obtaining antibodies to said protein and respective antibodies.

EFFECT: polypeptides can be used in realisation of methods stimulation of immune system, proliferation and development of hemopoietic cells in vitro and in vivo.

17 cl, 3 dwg, 21 tbl, 47 ex

FIELD: medicine.

SUBSTANCE: invention concerns virology and medicine area. The synthetic DNA-molecule coding protein L1 HPV45 is presented. Thus DNA-molecule was Codonum-optimised for high-level protein expression in a yeast cell. The given synthetic molecules can be used for reception of virus-like particles (VLP) HPV45 and for reception of vaccines and the pharmaceutical compositions containing VLP-particles HPV45.

EFFECT: effective immunologic prophylaxis of infections with papilloma virus due to neutralised antibodies and cellular immunity.

9 cl, 6 dwg, 8 ex

FIELD: medicine.

SUBSTANCE: human antibodies or their antigen-binding fragments are fully described, which specifically bind to human 4-1BB, and which provide for binding of human 4-1BB to human 4-1BB ligand. Antibody includes variable areas of light and heavy chains with amino-acid sequence, given in formula, and may in one of aspects represent IgG4 antibody. Invention presents polynucleotides, which code amino-acid sequences of heavy chain and light chain of monoclonal antibody. Pharmaceutical compositions are described for cancer treatment on the basis of monoclonal antibody or its fragment and method for treatment of disease in subject, which consists in introduction of therapeutically efficient amount of antibody to this subject.

EFFECT: antibodies of invention have agonistic activity and may be used for treatment or prevention of human diseases, such as cancer, infectious and autoimmune diseases.

10 cl, 24 dwg, 1 tbl, 3 ex

FIELD: chemistry.

SUBSTANCE: proposed is a recombinant single-strand trispecific antibody for treating tumours which express CEA. The said antibody consists of a series of three antibody fragments: anti-CEA-scFv, anti-CD3-scFv and VH CD28-antibody, linked by two intermediate linkers (intermediate linker Fc and intermediate linker HSA). If necessary, a c-myc-mark or (His)6-mark can be added at the C-end. Described is DNA, which codes the antibody, expression vector based on it and E.coli cell, containing the vector.

EFFECT: use of the invention is more beneficial in clinical use compared to bispecific antibodies and known trispecific antibodies, makes easier clearing and expression of an antibody, which can further be used in treating CEA-mediated tumours.

10 cl, 21 dwg, 11 ex

FIELD: chemistry, medicine.

SUBSTANCE: novel antibodies and fragments of human antibodies are bound with GDF-8 in a specific way and inhibit its activity in vitro and/or in vivo. On the basis of said invention pharmaceutical composition is created, which can be used for diagnostics, prevention or treatment of degenerative dysfunctions of muscle or bone or disorders of insulin metabolism.

EFFECT: extending range of arsenal of technical means used in treatment of diseases related to muscular, bone tissue or insulin metabolism.

FIELD: medicine.

SUBSTANCE: polypeptides include single-domain antibody against vWF, A1 domain of vWF, A1 domain of activated vWF, A3 domain of vWF, gp1b and/or collagen. Invention claims methods of obtaining indicated polypeptides, methods of coating devices applied in medical practice (e.g. in X-ray structural analysis, endoprosthetics) with indicated polypeptides.

EFFECT: obtainment of polypeptides for treatment of diseases requiring modulation of thrombocyte-mediated aggregation.

40 cl, 69 ex, 30 dwg, 32 tbl

FIELD: biotechnology.

SUBSTANCE: present invention relates to biotechnology and immunology. Proposed here is a polynucleotide, encoding a cyclic single-stranded tri-specific antibody. The antibody is directed against human ovarian carcinoma in vitro, has mass of approximately 84 kD and consists of three components: an antibody against human ovarian carcinoma cells, anti-CD3 antibody and anti-CD28 antibody, which are joined together by peptide interlinks such that, they form a cyclic antibody. Invented is an expression vector, containing a coding polynucleotide and versions of E.coli host cell based on the polynucleotide and expression vector.

EFFECT: use of the invention provides for a stable antibody molecule, optimum for activation of T-cells, which can be used in curing human ovarian carcinoma.

8 cl, 12 dwg

FIELD: biology; biotechnology.

SUBSTANCE: invention concerns biotechnology. It claims linking molecule represented by monoclonal antibody or its fragment capable of binding human NogoA polypeptide, human NogoA 623-640. Polynucleotide encoding the claimed molecule is presented. Expression vector including indicated polynucleotide is described. Host cell including indicated polynucleotide or vector is described. Pharmaceutical composition for treatment of diseases related to nerve reconstruction and containing indicated molecule is described.

EFFECT: obtainment of antibodies capable of binding NogoA 623-640.

9 cl, 1 dwg, 3 tbl, 7 ex

FIELD: medicine; bioengineering.

SUBSTANCE: are offered variants of antibodies, specifically recognizing two regions of peptide β-A4, characterised by amino-acid residual list. The first region of recognised peptide contains amino-acid sequence AEFRHDSGY or fragment thereof, while the second region contains amino-acid sequence VHHQKLVFFAEDVG or fragment thereof. Described are nucleic acid molecules coding molecules of antibodies, offered in invention, vectors and hosts containing specified nucleic acid molecules. Discovered are methods of antibodies production and optimisation, pharmaceutical compositions based on specified antibodies and method of production thereof, as well as antibodies-based set and various applications of antibodies. Invention application provides high-avidity antibodies of peptide β-A4 that can be applied for diagnostics of various peptide β-A4 mediated diseases.

EFFECT: efficient application of compositions.

29 cl, 15 dwg, 10 tbl, 16 ex

FIELD: biochemistry.

SUBSTANCE: invention refers to antibodies that link with CTGF. Antibodies, in particular, are directed to the areas of CTFG participating in different types of biological activity related to fibrosis. The invention also refers to the method of antibodies application in pharmaceutical compositions for the treatment of CTGF-related diseases including localised and systemic fibrotic diseases such as diseases of lungs, liver, heart, skin and kidneys, for neutralisation of biological activity of CTGF and method of treatment and prophylaxis of CTGF-related diseases. The invention covers polynucleotide sequence and its variants coding the specified antibody as well as the host cell and cell line № PTA-6006 (ATCC) producing the specified antibody.

EFFECT: use of the invention provides new specific means - antibodies which effectively neutralise certain types of CTGF activity in pathology and provide specificity and pharmacokinetic profile suitable for a therapeutic agent.

82 cl, 33 dwg, 4 tbl, 12 ex

FIELD: bioengineering.

SUBSTANCE: versions of the molecule binding CD45RO and CD45RB, and the anti-CD45RO and anti-CD45RB antibody are invented. In one of versions, the said molecule contains at least one antigen-binding site and includes the subsequently located hypervariable sites CDR1, CDR2 and CDR3. The molecule represents the humanised or monoclonal antibody. CDR1 has the amino acid sequence NYIIH, CDR2 has the amino acid sequence YFNPYNHGTKYNEKFKG and CDR3 has the amino acid sequence SGPYAWFDT. The molecule can additionally contain the subsequently located hypervariable sites CDR1', CDR2' and CDR3'. CDR1' has the amino acid sequence RASQNIGTSIQ, CDR2' has the amino acid sequence SSSESIS and CDR3' has the amino acid sequence QQSNTWPFT. In another version, the molecule contains both heavy and light chains where the amino acid sequences contain the corresponding CDR. The versions of the corresponding coding polynucleotide are disclosed; expression vector and based on it expression system. The host cell is disclosed basing on the expression system. The application of the molecule in treatment of autoimmune diseases, graft rejection, psoriasis, intestine inflammatory disease and allergy is described. The pharmaceutical composition for the said application is disclosed.

EFFECT: enables immunosuppressant induction; inhibiting T-cell response and primary lymphocyte response in mixed lymphocyte culture (MLC); prolongs survival period in mice with severe combined immunodeficiency SCID.

20 cl, 5 dwg, 2 tbl, 8 ex

FIELD: medicine.

SUBSTANCE: described humanised and chimeric CD20 antibodies are designed for treatment of CD20-positive malignant and autoimmune diseases. Antibody is effective with respect to depletion of B-cells of mammals in vivo, contains in variable region of H-chain of CDR3- sequence from antibody to human CD20 and practically all remains of consensus frame region (FR) of human H-chain of subgroup 111. According to invention antibody is used in composition or product, binding CD20. Besides, antibody is used for apoptosis induction, treatment of CB20-positive cancer, autoimmune disease, and rheumatic arthritis. Invention contains nucleic acid (NA) coding antibody, expression vector containing specified NA, and host cell producing recombinant antibody, as well as method of specified antibody production. According to invention antibodies are characterised by minimum antigenicity or no antigenicity at all, that enables to use them for continuous treatment overcoming limits of existing therapeutic compositions application.

EFFECT: enables to use for continuous treatment.

83 cl, 32 dwg, 12 tbl, 16 ex

FIELD: medicine.

SUBSTANCE: there are disclosed polypeptide variants containing Fc-areas IgG, having amino acid modifications providing changed effector functions Fc in specified polypeptides. There is disclosed composition for antibody targeting on antigen, containing the specified polypeptide. There is described method for preparing the specified polypeptide. Also, there are disclosed the methods for treating V-cell tumour or a malignant disease characterised by V-cell expression of CD20, treating chronic lymphocytic leukosis, relieving the symptoms of the V-cell controlled autoimmune disease, treating a angiogenesis-associated disorder, treating HER2-expressing cancer, treating LFA-1-mediated involvement, treating IgE-mediated involvement wherein specified methods imply introduction to the patient of the therapeutically effective amount of said polypeptide.

EFFECT: higher clinical effectiveness.

63 cl, 6 ex, 13 dwg, 10 tbl

Up!