CN106632673A

CN106632673A - Anti-PG (prostaglandin)E2 monoclonal antibody and application thereof

Info

Publication number: CN106632673A
Application number: CN201510723595.8A
Authority: CN
Inventors: 钮晓音
Original assignee: Individual
Current assignee: Individual
Priority date: 2015-11-01
Filing date: 2015-11-01
Publication date: 2017-05-10

Abstract

The invention discloses an anti-PG (prostaglandin)E2 monoclonal antibody and an application thereof and provides composition containing the anti-PGE2 monoclonal antibody or a fragment of the anti-PGE2 monoclonal antibody, and a method for using the anti-PGE2 monoclonal antibody or a fragment of the anti-PGE2 monoclonal antibody. The anti-PGE2 monoclonal antibody or the fragment of the anti-PGE2 monoclonal antibody contains CDR sequences SDGKTY(SEQ ID NO:1), LVS(SEQ ID NO:2) and WQGTHFPWT(SEQ ID NO:3) in variable regions of a light chain and CDR sequences SRGDWFHA(SEQ ID NO:4), AQTHGSQS(SEQ ID NO:5) and QKISPEQFHGAYS(SEQ ID NO:6) in variable regions of a heavy chain. The anti-PGE2 monoclonal antibody or the fragment of the anti-PGE2 monoclonal antibody can block binding of PGE2 with an EP receptor of the PGE2, thereby blocking intracellular signal transfer of the PGE2. The anti-PGE2 monoclonal antibody or the fragment of the anti-PGE2 monoclonal antibody is beneficial to treatment of autoimmune diseases and can inhibit differentiation of Tfh cells and down-regulate inflammatory reaction.

Description

Anti- PGE2 monoclonal antibodies and its use

Technical field

The invention belongs to medical biotechnology and humanized antibody research field.The present invention relates to resist Prostaglandin E2（PGE2）Antigenic determinant there is the monoclonal antibody molecule and the compositionss comprising the antibody molecule and its fragment of specific recognition.The invention further relates to purposes of the anti-Prostaglandin E2 antibody in treatment autoimmune disease.

Background technology

Prostaglandin（Prostaglandin, PG）It is the class unsaturated fatty acid composition, active substance with various physiological actions being present in animal and human body.Find that it is present in the seminal fluid of people earliest, think that this material is discharged by prostate at that time, thus name as prostaglandin.Prostaglandin can be divided into the types such as A, B, C, D, E, F, G, H, I by structure, and wherein PGE2 is a most studied type.

PGE2 has very strong adjustment effect to cell itself or to the physiological activity of adjacent cells.Prostaglandin Endocrine, reproduction, digestion, blood breathing, cardiovascular, urinary system and nervous system have effect.

PGE2 can induce inflammation, promote local vascular dilation, capillary permeability to increase, and cause the symptom such as red, swollen, pain, heat.

The content of the invention

The present invention is, based on one plant of obtained anti-prostaglandin E2 mouse monoclonal antibody 3G11, by clone, identification and the analysis of gene structure, it is determined that its CDR region sequence, to construct corresponding human mouse chimeric antibody and its eukaryotic expression vector.

The present invention have developed the specific antibody for prostaglandin E2 micromolecular compounds, and the epi-position that it is combined is located on prostaglandin E2.

Our in vitro study finds that Prostaglandin E2 can promote Tfh cell growths, promotes inflammation to be formed.

After using anti-PGE2 monoclonal antibodies blocks, the propagation of Tfh cells is substantially suppressed.

Anti- PGE2 monoclonal antibodies suppress the Mechanism Study of inflammatory reaction, and theoretical basiss are provided in the application of clinical treatment autoimmune disease for this antibody, and the immunization therapy based on this antibody also can have broad application prospects.

Therefore, the invention mainly relates to the following aspects：

First aspect, the present invention relates to a kind of anti-prostaglandin E2 monoclonal antibodies, wherein described antibody is made up of the light chain containing 3 light chain variable districts and the heavy chain containing 3 weight chain variable districts, wherein the aminoacid sequence of 3 light chain variable districts CDR is SDGKTY (SEQ ID NO：1)、LVS(SEQ ID NO ：2) with WQGTHFPWT (SEQ ID NO：And weight chain variable district CDR sequence SRGDWFHA (SEQ ID NO 3)：4)、AQTHGSQS (SEQ ID NO ：5) with QKISPEQFHGAYS (SEQ ID NO：6).It is known in the art that the binding specificity and affinity of antibody are mainly determined by CDR sequences, the aminoacid sequence in non-CDR regions can be changed easily according to ripe, known existing every technology and obtain the variant with similar biological activity.Variant monoclonal antibody with the identical CDR sequences of above-mentioned CDR sequences according to the present invention, because it has and the identical CDR sequences of the chimerization antibody of people Mus of the present invention, therefore with similar biological activity.

Second aspect, the present invention relates to the fragment of anti-prostaglandin E2 monoclonal antibodies, the wherein fragment are selected from F (ab ')₂, Fab ', Fab, Fv, scFv and antibody atom composition group.And the fragment contains the CDR of CDR and 3 weight chain variable district of 3 light chain variable districts, its light chain variable district cdr amino acid sequence is SDGKTY (SEQ ID NO：1)、LVS(SEQ ID NO ：2) with WQGTHFPWT (SEQ ID NO：And weight chain variable district CDR sequence SRGDWFHA (SEQ ID NO 3)：4)、AQTHGSQS (SEQ ID NO ：5) with QKISPEQFHGAYS (SEQ ID NO：6).

The third aspect, the present invention relates to a kind of method for treating disease, wherein the disease is selected from group consisting of：Autoimmune disease, such as：Rheumatoid arthritiss, systemic lupus erythematosus (sle), multiple sclerosis, systemic vasculitiss, autoimmune liver disease, pemphiguss, scleroderma, dermatomyositiss, mixed connective tissue disease, autoimmune hemolytic anemia, Autoimmune Thyroid Diseases, ulcerative colitiss, CrohnShi is sick, psoriasises, drying property syndrome, pernicious anemia, neonatal hemolytic, autoimmune hemolytic anemia, idiopathic thrombocytopenic purpura, insulin dependent diabetes mellitus (IDDM), ankylosing spondylitises, scleroderma, myasthenia graviss, polyarteritis nodosa, Wegener granulomatosis, subacute bacterial endocarditiss etc.；Immunologic rejection disease, such as host-versus-graft reaction, graft versus host disease, transfusion-associated graft versus host disease, anaphylactic reaction.

Claims

1. a kind of monoclonal antibody, single-chain antibody or its antibody fragment and combinations thereof, it is included comprising light chain variable district CDR (complementary determining region) sequence SDGKTY (SEQ ID NO：1)、LVS(SEQ ID NO ：And WQGTHFPWT 2) (SEQ ID NO ：And weight chain variable district CDR sequence SRGDWFHA (SEQ ID NO 3)：4)、AQTHGSQS (SEQ ID NO ：And QKISPEQFHGAYS 5) (SEQ ID NO ：6).

2. according to claim 1, wherein anti-PGE2 monoclonal antibodies or its fragment are exposed antibody or its fragment.

3. according to claim 1, wherein the variable region of anti-PGE2 monoclonal antibodies forms compositionss with the sequence of human antibody IgG1, IgG2, any one of them constant region of IgG3, IgG4, IgA, IgM, IgE, IgD.

4. compositionss according to claim 1, wherein the antibody fragment is selected from F (ab ')₂, Fab ', Fab, Fv, scFv and antibody atom composition group.

5. compositionss according to claim 1, wherein the anti-PGE2 monoclonal antibodies or its fragment are coupled to form immune conjugate with least one diagnostic agent and/or therapeutic agent.

6. compositionss according to claim 5, wherein the therapeutic agent is selected from group consisting of：Exposed antibody, cytotoxic agent, medicine, radionuclide, boron atom, immunomodulator, heliosensitivity therapeutic agent, immune conjugate, oligonucleotide and hormone.

7. a kind of method for treating disease, it includes using anti-PGE2 monoclonal antibodies or its fragment or compositionss to experimenter, wherein the anti-PGE2 monoclonal antibodies or its fragment include light chain variable district CDR sequence SDGKTY (SEQ ID NO：1)、LVS(SEQ ID NO ：And WQGTHFPWT 2) (SEQ ID NO ：And weight chain variable district CDR sequence SRGDWFHA (SEQ ID NO 3)：4)、AQTHGSQS (SEQ ID NO ：And QKISPEQFHGAYS 5) (SEQ ID NO ：6).

8. method according to claim 7, wherein the disease is selected from group consisting of：Rheumatoid arthritiss, systemic lupus erythematosus (sle), multiple sclerosis, systemic vasculitiss, autoimmune liver disease, pemphiguss, scleroderma, dermatomyositiss, mixed connective tissue disease, autoimmune hemolytic anemia, Autoimmune Thyroid Diseases, ulcerative colitiss, CrohnShi is sick, psoriasises, drying property syndrome, pernicious anemia, neonatal hemolytic, autoimmune hemolytic anemia, idiopathic thrombocytopenic purpura, insulin dependent diabetes mellitus (IDDM), ankylosing spondylitises, scleroderma, myasthenia graviss, polyarteritis nodosa, Wegener granulomatosis, subacute bacterial endocarditiss etc., host-versus-graft reaction, graft versus host disease, transfusion-associated graft versus host disease, anaphylactic reaction.