CN103923183A - Interleukin-6 polypeptide inhibitor and its application - Google Patents

Abstract

The invention relates to the field of medicines, and concretely relates to a polypeptide capable of inhibiting interleukin-6 and preventing and treating rheumatoid arthritis. The sequence of the polypeptide is TEDGGGTTPNRRGD which is a brand new sequence. The polypeptide can be used for treating the rheumatoid arthritis and has potential new drug exploitation values.

Description

Interleukin-6 peptide inhibitor and application thereof

technical field:

The present invention relates to pharmaceutical field, be specifically related to the interleukin-6 peptide inhibitor for preventing and treat or prevent rheumatoid arthritis.

background technology:

Rheumatoid arthritis (rheumatoid arthritis, RA) is one of clinical modal inflammatory arthropathy and main disability-causing factor.Be about 0.5%-1.0% in the whole world, the morbidity of RA is about 0.4% in China.RA can betide any age, and with advancing age, sickness rate also increases thereupon.Women is 45-55 year at the age occurred frequently, and sex and RA onset relation are close, men and women's the about 1:3 of ratio.RA is the chronic systemic inflammatory disease that a kind of cause of disease not yet understands, taking pathology outside chronic, symmetry, many synovial joints inflammation and joint as main clinical manifestation, belongs to autoimmunization inflammatory diseases.Often taking hand or wrist pain and swelling (the particularly swelling at wrist back), as onset symptoms, symptom continues not alleviate patient, though common symptomatic treatment can relief of symptoms, usually because medication is irregular or in shortagely cause symptom repeatedly.When disease progression, can there is deadlock in obvious morning, conventionally can reach more than 1 hour, and constantly increase the weight of; There is certain joint function disturbance simultaneously.

The cause of disease of rheumatoid arthritis and pathogenesis are not yet completely clear, and its basic pathology feature is vasculitis and synovitis.Intraarticular synovial membrane blood vessel hyperplasia, forms pannus, causes synovial membrane to thicken, and oozes out and increases, and secretion cytokine profiles, invades cartilage and cause bone damage.Its flesh chamber, ligament, stndon sheath and muscle etc. are around organized also to all erodables, thereby affect stablizing of joint, easily joint deformity occurs and occur dysfunction.Vasculitis also can be invaded the each organs and tissues in the whole body, forms systemic disease.

The medicine for the treatment of RA is divided into two large classes at present: control symptom medicine and control disease medicine.Due to RA etiology unknown, can be rated as therefore there is no now the medicine of controlling disease.The antirheumatic of controlling symptom is divided into four classes: one, non-steroidal anti-inflammatory drugs, be commonly referred to a line medicine, and this class medicament categories is various, and domestic market has reached tens of kinds.Two, steroid hormone, hormone is an extraordinary pain stop and anti-inflammation medicine, but long-term use separately can not improve the state of an illness, bring on the contrary many side effects, hormone is made transitionality before medication onset with being fine slowly as two wires, but consumption is little, and the time is unsuitable long.Heavier in the state of an illness, with the patient of extra-articular manifestation, short impacts, and necessary while combining the treatment of two wires medicine.Three, act on slowly antirheumatic, be commonly referred to two wires medicine, so-called slow effect cartridge bag is drawn together antimalarial drug, golden salt, Trolovol and willow nitrogen sulfapyridine, and their treatment RA onsets are slow, and long term has certain mitigation to the RA state of an illness, therefore the feelings of also pretending illness are improved medicine.Four, immunosuppressor: commonly use and have Rheumatrex, endoxan, azathioprine, trypterygine, Stem of Orientoine etc.

In the biotechnological formulation for the treatment of RA, studying clinically more is exactly the antagonist of interleukin-6 (Il-6).Il-6 is the T cell of activation and the lymphokine that inoblast produces.Can make B cell precursor become the cell that produces antibody; Collaborative with G CFS, can promote growth and the differentiation of original bone marrow-derived cells, strengthen the cracking function of natural killer cell, in RA pathogenic process, play an important role.The effect of Il-6 antagonist is antagonism Il-6 specifically, suppresses the disease progression of RA.The Il-6 antagonist of approved listing is at present mainly Actemra (holder pearl monoclonal antibody).Because this medicine is monoclonal antibody, exist molecular weight larger, and produce the shortcomings such as antigen-reactive, greatly limit it in clinical use.

summary of the invention:

The present invention seeks to the feature for existing Il-6 antagonist, design the Il-6 polypeptide antagonist that a kind of molecular weight is little, can avoid producing antigen-reactive, be more applicable for clinical application.

Technical solution of the present invention is to provide a kind of interleukin-6 peptide inhibitor 2, and its sequence is TEDGGGTTPNRRGD, and the application in medicine for treating rheumatoid arthritis in treatment or prevention.

useful result:

Interleukin-6 peptide inhibitor 2 sequence TEDGGGTTPNRRGD in the present invention, can suppress interleukin-6 by target, suppress the physiology of Tumor Necrosis Factor Receptors generation or the effect of pathology, reach the effect of prevention or treatment rheumatoid arthritis.

Contriver knows that through great many of experiments interleukin-6 peptide inhibitor 2 can suppress the development of adjuvant type rat kind rheumatic arthritis and DBA/1 mouse collagen type rheumatoid arthritis, in body, experimental results show that and there is the arthritic effect of significant treatment similar rheumatism type, and few side effects, the few cost of consumption.The interleukin-6 peptide inhibitor 2 that the present invention's design is described is scientific, rational, feasible and effective, can be as treatment or prevention medicine for treating rheumatoid arthritis.

Embodiment

Embodiment 1

Interleukin-6 peptide inhibitor 2 is immanoprotection action in collagen-induced mouse arthritis animal model

Build collagen type mouse arthritis animal model, the therapeutic action of research interleukin-6 peptide inhibitor 2 to mouse Collagen-induced Arthritis (collagen induced arthritis, CIA).Adopt mouse as animal subject, 90 of SPF level DBA/1 mouse (are provided by Shanghai western pul-Bi Kai laboratory animal company limited (Sino-British SIPPR Lab. Animal Ltd), animal production licence number: SCXK (Shanghai) 2008-0016), male, age in 7-8 week, body weight is 18-22 g, be divided at random 6 groups, it is respectively Normal group, model control group, low middle high 3 the dosage groups of peptide inhibitor 2 (0.4,0.8,1.6 mg/kg) and positive drug control group (methotrexate 2 mg/kg).Except normal group, within the 0th day, each experimental group is set up mouse CIA model, and method is chicken cartilage II Collagen Type VI (c II) becomes 4 mg/ml solution with 0.1 mol/l acetate dissolution, in 4 DEG C of refrigerator overnight.Experiment was fully emulsified with complete Freund's adjuvant (CFA) equal-volume containing 4 mg/ml Myeobaeterium tuberculosis strain H37Rv with II Collagen Type VI the same day, after DBA/1 mouse anesthesia, in every of its afterbody intracutaneous injection emulsifying agent, 50 μ l carry out sensitization, carry out again immune after fully emulsified with incomplete Freund's adjuvant (IFA) equal-volume with the II Collagen Type VI (c II) of 4 mg/ml after 21 d with the emulsifying agent of same dose in afterbody.Modeling the 30th d plays subcutaneous injection administration: 3 dosage groups (0.4,0.8,1.6 mg/kg) of polypeptide 2: every day twice, continuous 10 days; Positive drug control group (methotrexate 2 mg/kg): every five days once, continuous 3 times; Normal group and model control group (physiological saline): continuous 10 days.Respectively within after modeling the 21st day to the 70th day every 3 days, weighing in, joint scoring, detect left and right metapedes ankle diameter respectively and observe medicine to the arthritic impact of mouse collagen type.

Result: after modeling, mouse is compared with normal mouse, latter the 27th day of immunity, CIA mouse foot pawl redness, arthritis index scoring is increased, and model group 45-60 days is swelling peak, and model group started body weight from the 35th day to be increased hardly, and the later stage also has decline slightly.Interleukin-6 peptide inhibitor 2 immanoprotection action concrete outcome in collagen-induced mouse arthritis animal model is as follows:

The impact of interleukin-6 peptide inhibitor 2 on the left whole pawl swelling of mouse collagen type sacroiliitis, positive controls, the left back ankle diameter of the high, medium and low dosage group of interleukin-6 peptide inhibitor 2 and model group comparison, all have utmost point significant difference (P<0.01) experimental result to have statistical significance.The impact of interleukin-6 peptide inhibitor 2 on the right whole pawl swelling of mouse collagen type sacroiliitis, positive controls, the right back ankle diameter of the high, medium and low dosage group of interleukin-6 peptide inhibitor 2 and model group comparison, all have utmost point significant difference (P<0.01), experimental result has statistical significance.The impact of interleukin-6 peptide inhibitor 2 on collagen type sacroiliitis mouse clinical score, the basic, normal, high dosage group of polypeptide 2 four limbs are marked significantly lower than model control group (P < 0.01), with relatively utmost point significant difference of model control group, experimental result has statistical significance.Interleukin-6 peptide inhibitor 2 affects collagen type sacroiliitis Mouse Weight, the high, medium and low dosage group of polypeptide 2 body weight is significantly higher than model control group (P < 0.01), with relatively utmost point significant difference of model control group, experimental result has statistical significance.

Conclusion: interleukin-6 peptide inhibitor 2 has therapeutic action to mouse collagen type sacroiliitis.

Embodiment 2

Interleukin-6 peptide inhibitor 2 is to immanoprotection action in adjuvant type rat arthritis animal model

Build adjuvant type rat arthritis animal model, the therapeutic action of research interleukin-6 peptide inhibitor 2 to adjuvant-induced arthritis (Adjuvant arthritis, AA) rat.Adopt rat as animal subject, 90 of SPF level SD rats (are provided by Shanghai western pul-Bi Kai laboratory animal company limited (Sino-British SIPPR Lab. Animal Ltd), animal production licence number: SCXK (Shanghai) 2008-0016), male, body weight is 140 g-160 g, be divided at random 6 groups, it is respectively Normal group, model control group, low middle high 3 the dosage groups (0.2 of interleukin-6 peptide inhibitor 2,0.4,0.8 mg/kg) and positive drug control group (methotrexate 1 mg/kg).Except normal group, within the 0th day, each experimental group is set up rat AA model, and method is that the injection of wasting time fully afterwards in the left side of rat causes experimental animal model of CFA induced adjuvant arthritis in rats containing mycobacterium tuberculosis (H37RA, 10 mg/ml) complete Freund's adjuvant 0.08 ml of deactivation.Modeling plays subcutaneous injection administration on the 10th day: 3 dosage groups (0.2,0.4,0.8 mg/kg) of polypeptide 2: every day twice, continuous 10 days; Positive drug control group (methotrexate 1 mg/kg): every five days once, continuous 3 times; Normal group and model control group (physiological saline): continuous 10 days.Respectively at after modeling the 8th, 11,14,17,20,23 and 26 days, weigh in, joint scoring, detect left and right metapedes ankle diameter respectively and observe medicine to the arthritic impact of rat adjuvant type.

Result: after modeling, rat is compared with normal rat, the left back foot of AA rat can produce rapidly primary sacroiliitis, occurs obvious swelling, and with festering; After about 10 d of right back foot, start to occur post-traumatic arthritis, the score value of scoring increases gradually; Ear's blood vessel hyperplasia is obvious simultaneously, obviously red and swollen; There is swelling in caudal articular process.Interleukin-6 peptide inhibitor 2 is immanoprotection action in adjuvanticity rat arthritis animal model, and concrete outcome is as follows:

The impact of interleukin-6 peptide inhibitor 2 on rat primary sacroiliitis foot pawl swelling, the left back ankle diameter of basic, normal, high dosage group and the model group comparison of rat positive controls, interleukin-6 peptide inhibitor 2, have utmost point significant difference (P<0.01); The left back ankle diameter of the basic, normal, high dosage group of interleukin-6 peptide inhibitor 2 and model group comparison, all have significant difference (P<0.05), and experimental result has statistical significance.The impact of interleukin-6 peptide inhibitor 2 on rat post-traumatic arthritis foot pawl swelling, rat positive controls, the right back ankle diameter of the basic, normal, high dosage group of interleukin-6 peptide inhibitor 2 and model group comparison, have significant difference (P<0.05).The impact of interleukin-6 peptide inhibitor 2 on adjuvant type rats with arthritis clinical score, the basic, normal, high dosage group of polypeptide 2 four limbs are marked significantly lower than model control group (P < 0.05), all have statistical significance with model control group comparing difference.Interleukin-6 peptide inhibitor 2 is on the impact of adjuvant type rats with arthritis body weight, and the high, medium and low dosage group of polypeptide 2 body weight is significantly higher than model control group (P < 0.05), all has statistical significance with model control group comparing difference.

Conclusion: interleukin-6 peptide inhibitor 2 has therapeutic action to AA rat arthritis.

SEQUENCE LISTING

<110> Nantong Chengxin Amino Acid Co., Ltd.

<120> interleukin-6 peptide inhibitor 2 and application thereof

<130>

<160> 1

<170> PatentIn version 3.3

<210> 1

<211> 14

<212> PRT

<213> artificial sequence

<400> 1

Thr Glu Asp Gly Gly Gly Thr Thr Pro Asn Arg Arg Gly Asp

1 5 10

Claims (2)

1. interleukin-6 peptide inhibitor 2, is characterized in that its sequence is TEDGGGTTPNRRGD.

2. interleukin-6 peptide inhibitor 2, the application in treatment or prevention medicine for treating rheumatoid arthritis.