CN109730993A - Application of the indolepopionic acid in preparation prevention and treatment rheumatoid arthritis drug - Google Patents
Application of the indolepopionic acid in preparation prevention and treatment rheumatoid arthritis drug Download PDFInfo
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Abstract
The present invention provides application of the indolepopionic acid in preparation prevention and treatment rheumatoid arthritis drug, are related to the treatment technology field of rheumatoid arthritis disease.Indolepopionic acid can significantly inhibit the occurrence and development of experimental arthritis, and arthroncus symptom is effectively relieved.Indolepopionic acid can inhibit proinflammatory cytokines Th1 and folliculus helper T lymphocyte (Follicular helper T cell, Tfh) and the ratio for raising anti-inflammatory cells regulatory T cells (T regulatory, Treg).Indolepopionic acid can also reduce proinflammatory factor IL-1 β in serum, IL-6, the expression of IL-17A and autoantibody anti-CII (anti-type II collage antibody), and raise the secretion with the beta-endorphin (β-endorphin) of analgesic activity.Indolepopionic acid has anti-inflammatory and immunosuppressive action, can correct the intracorporal immunocyte imbalance state of biology, and then achievees the purpose that prevent and treat rheumatoid arthritis.
Description
Technical field
The present invention relates to the treatment technology fields of rheumatoid arthritis disease, and in particular to indolepopionic acid prevents and treats class in preparation
Application in rheumathritis drug.
Background technique
Rheumatoid arthritis lesion mainly invades joint, the general recurrent exerbation of joint symptoms, and destruction of joint is got worse, most
Lead to the dysfunction and deformity of varying degree afterwards.In addition to joint, skin rheumatoid nodules, arteritis, pericarditis, sclerotitis,
Lymphnoditis, hepatosplenomegaly, neuropathy etc. are quite a few to see.The illness rate of China's rheumatoid arthritis is 0.28-0.41%, is suffered from
Person up to as many as 5,000,000, are up to 75% without regular treatment disability rate.The Second National disabled person that China was carried out in 2006
In sample investigation, crippling arthropathy arranges the 2nd (20.1%) in the 21 class causes of disease for leading to physical disabilities, is only second to the cerebrovascular
Sick (20.6%), and wherein 79.4% disable for RA.Systemic loupus erythematosus SLE morbidity's initial stage shows as low-heat, face more
The clinical symptoms such as portion's fash, out of strength, alopecia, arthralgia, serious person will appear the neurological symptoms such as twitch, syncope, and advanced stage is more
Development is chronic nephritis even kidney failure.Illness rate of the disease in China is about 70/,100,000 people, and number of patients is up to 900,000,5 years
Survival rate only has 50-85%, and poor prognosis seriously threatens the quality of life of patient, brings heavy spirit to patient and society
And financial burden.
The cause of disease and pathogenesis of rheumatoid arthritis are still not clear, and treatment is only limitted to respite and improves the state of an illness, no
The occurrence and development of disease can fundamentally be controlled.Therefore, the pathogenesis of rheumatoid arthritis is probed into comprehensively and in depth, is found
The early diagnosis index and effective therapy target of specificity have important theory significance and clinical value.Currently, class wind
Wet arthritic treatment is remained as using immunosuppressor, hormone and (or) biological agent.However, excessively using hormone, being immunized
The inhibition such as inhibitor and biological agent immune response can not achieve the purpose that disease is effectively relieved.Therefore novel tune is found
Control immune imbalance and the treatment method for rebuilding immune tolerance are particularly important.
Summary of the invention
The object of the present invention is to provide application of the indolepopionic acid in preparation prevention and treatment rheumatoid arthritis drug.Indoles
Propionic acid can significantly inhibit the occurrence and development of experimental arthritis, and arthroncus symptom is effectively relieved.Indolepopionic acid can inhibit proinflammatory
Cell Th1 and folliculus helper T lymphocyte (Follicular helper T cell, Tfh) and to raise anti-inflammatory cells regulatory T thin
The ratio of born of the same parents (T regulatory, Treg), can also reduce proinflammatory factor IL-1 β, IL-6, IL-17A and autoantibody in serum
The expression of anti-CII (anti-type II collage antibody), and raise the beta-endorphin (β-with analgesic activity
Endorphin secretion).Indolepopionic acid has anti-inflammatory and immunosuppressive action, and it is unbalance can to correct the intracorporal immunocyte of biology
State, and then achieve the purpose that prevent and treat rheumatoid arthritis.
The present invention provides application of the indolepopionic acid in preparation prevention and treatment rheumatoid arthritis drug.
Preferably, the drug further includes indolepopionic acid pharmaceutically acceptable auxiliary material.
The present invention also provides indolepopionic acids to lower in histoorgan in the drug of the expression of proinflammatory cytokines Th1 in preparation
Using.
The present invention also provides indolepopionic acids to reduce answering in the drug of the expression of Tfh cell in histoorgan in preparation
With.
The present invention also provides indolepopionic acids to raise answering in the drug of the expression of Treg cell in histoorgan in preparation
With.
Preferably, the histoorgan includes joint lymph node, lymphonodi mesenterici or spleen.
The present invention also provides indolepopionic acids to reduce the application in serum in the drug of the expression of proinflammatory factor in preparation.
Preferably, the proinflammatory factor includes IL-1 β, IL-6 and IL-17A.
The present invention also provides application of the indolepopionic acid in preparation up-regulation serum in the drug of the secretion of beta-endorphin.
The present invention provides application of the indolepopionic acid in preparation prevention and treatment rheumatoid arthritis drug, indolepopionic acid can pass through
Inhibit proinflammatory cytokines Th1 and folliculus helper T lymphocyte and the ratio for raising anti-inflammatory cells regulatory T cells, can also reduce serum
Middle proinflammatory factor IL-1 β, IL-6, IL-17A and autoantibody anti-CII (anti-type II collage antibody)
Expression, and raise have analgesic activity beta-endorphin (β-endorphin) secretion.Indolepopionic acid has anti-inflammatory and immune
Inhibiting effect can correct the intracorporal immunocyte imbalance state of biology, and then achieve the purpose that prevent and treat rheumatoid arthritis.
Detailed description of the invention
Fig. 1 is that indolepopionic acid content substantially reduces result in rheumatoid arthritis patients serum；
Fig. 2 is indolepopionic acid Inhibition test arthritis occurrence and development result；
Fig. 3 is the ratio that indolepopionic acid regulates and controls experimental arthritis mouse lymphocyte subgroup；
Fig. 4 is the expression that indolepopionic acid regulates and controls autoantibody and inflammatory mediator.
Specific embodiment
The present invention provides application of the indolepopionic acid in preparation prevention and treatment rheumatoid arthritis drug.The present invention is to the Yin
The source of diindyl propionic acid is not particularly limited, using conventional commercial product.In the present invention, the drug further includes indoles third
The pharmaceutically acceptable auxiliary material of acid.The present invention is not particularly limited the content of indolepopionic acid in the drug, the present invention couple
The dosage form of the drug is not particularly limited, indolepopionic acid pharmaceutically acceptable dosage form.
The present invention also provides indolepopionic acids to lower in histoorgan in the drug of the expression of proinflammatory cytokines Th1 in preparation
Using.In the present invention, the organ preferably includes joint lymph node, lymphonodi mesenterici or spleen.The present invention is to the Yin
The source of diindyl propionic acid is not particularly limited, using conventional commercial product.In the present invention, the drug further includes indoles third
The pharmaceutically acceptable auxiliary material of acid.The present invention is not particularly limited the content of indolepopionic acid in the drug, the present invention couple
The dosage form of the drug is not particularly limited, indolepopionic acid pharmaceutically acceptable dosage form.
The present invention also provides indolepopionic acids to reduce answering in the drug of the expression of Tfh cell in histoorgan in preparation
With.In the present invention, the histoorgan preferably includes joint lymph node, lymphonodi mesenterici or spleen.The present invention is to described
The source of indolepopionic acid is not particularly limited, using conventional commercial product.In the present invention, the drug further includes indoles
Propionic acid pharmaceutically acceptable auxiliary material.The present invention is not particularly limited the content of indolepopionic acid in the drug, the present invention
The dosage form of the drug is not particularly limited, indolepopionic acid pharmaceutically acceptable dosage form.
The present invention also provides indolepopionic acids to raise answering in the drug of the expression of Treg cell in histoorgan in preparation
With.In the present invention, the histoorgan preferably includes joint lymph node, lymphonodi mesenterici or spleen.The present invention is to described
The source of indolepopionic acid is not particularly limited, using conventional commercial product.In the present invention, the drug further includes indoles
Propionic acid pharmaceutically acceptable auxiliary material.The present invention is not particularly limited the content of indolepopionic acid in the drug, the present invention
The dosage form of the drug is not particularly limited, indolepopionic acid pharmaceutically acceptable dosage form.
The present invention also provides indolepopionic acids to reduce the application in serum in the drug of the expression of proinflammatory factor in preparation.?
In the present invention, the proinflammatory factor preferably includes IL-1 β, IL-6 and IL-17.The present invention does not have the source of the indolepopionic acid
Particular determination, using conventional commercial product.In the present invention, the drug further includes that indolepopionic acid is pharmaceutically subjected to
Auxiliary material.The present invention is not particularly limited the content of indolepopionic acid in the drug, and the present invention does not have the dosage form of the drug
There is a particular determination, indolepopionic acid pharmaceutically acceptable dosage form.
The present invention also provides application of the indolepopionic acid in preparation up-regulation serum in the drug of the secretion of beta-endorphin.This
Invention is not particularly limited the source of the indolepopionic acid, using conventional commercial product.In the present invention, the drug
It further include indolepopionic acid pharmaceutically acceptable auxiliary material.The present invention does not have special limit to the content of indolepopionic acid in the drug
Fixed, the present invention is not particularly limited the dosage form of the drug, indolepopionic acid pharmaceutically acceptable dosage form.
Combined with specific embodiments below to indolepopionic acid of the present invention in preparation prevention and treatment rheumatoid arthritis drug
Application be further described in detail, technical solution of the present invention includes but is not limited to following embodiment.
Embodiment 1
Indolepopionic acid content substantially reduces in rheumatoid arthritis patients serum
Implementation method
(1) standard items configure: appropriate IPA (sigma) powder is dissolved in methanol solution, and being configured to concentration is 10mmol/mL's
Storage solutions.
(2) content of gas chromatography-mass spectrometry (GC-MS) detection indolepopionic acid: acquisition rheumatoid arthritis patients
The peripheral blood of (RA, n=60) and normal healthy controls crowd (HC, n=40) separate serum.It takes about 70 μ L in 1.5mLEP pipe, adds
Enter 210 μ L of acetonitrile shallow lake liquid, vortex 45s under 4 degrees Celsius, with 16000g/min, is centrifuged 10min, takes supernatant 100ul, sample introduction 20
μL.It is with PeakView1.2 software that the IPA in sample is qualitative referring to IPA standard items database, with MultiQuant2.1 to fixed
Property IPA it is quantitative, calculate respective concentration with standard curve.The result is shown in Figure 1.
Result of study
Fig. 1 is the results show that indolepopionic acid (IPA) content substantially reduces in RA patients serum, indoles third in Healthy Human Serum
The content of acid is 1689 ± 306.3nM, and the content of indolepopionic acid is only 656.9 ± 60.26nM in RA patients serum.Due to Yin
Diindyl propionic acid has the function of anti-inflammatory and inhibits immune response, prompts indolepopionic acid that may have protection to make in RA pathogenic process
With.
Embodiment 2
Indolepopionic acid Inhibition test arthritis occurrence and development
Implementation method
(1) it constructs collagen-induced property joint (CIA) model: 1) dissolving collagen, II Collagen Type VI of ox (C II) is dissolved in 0.1mol
In glacial acetic acid, II final concentration of C is adjusted to 4mg/mL, 4 DEG C overnight.2) collagen is emulsified, by the C II being completely dissolved and isometric Freund
Freund's complete adjuvant (CFA) mixing, keeps the two fully emulsified, the collagen emulsion of final concentration of 2mg/mL is made.3) initial immunity, mouse
Root of the tail portion unhairing, multiple intradermal injections total amount are the collagen emulsion of 200 μ g.4) booster immunization will the 21st day after initial immunity
II Collagen Type VI is dissolved in 0.1mol glacial acetic acid (concentration 4mg/mL), isometric incomplete Freund's adjuvant (IFA) is added, collagen is made
Emulsion (final concentration 2mg/mL).Root of the tail portion multiple intradermal injections total amount is the collagen emulsion of 100 μ g.
(2) indolepopionic acid intervention: 1) Primary preventive intervention: 3 weeks before arthritic start to be administered, and are administered once, give every other day
Prescription formula is stomach-filling, and grouping situation is as follows: disease control group (CIA), low dosage indolepopionic acid group (IPA-20mg/kg), high agent
It measures indolepopionic acid group (IPA-40mg/kg).2) therapeutic intervention: starting to be administered after the onset of mouse arthritis, once every other day, administration
Mode is stomach-filling, dosage 20mg/kg.
(3) Arthritis Incidence and arthritis score record: 1) Arthritis Incidence: (exempt from for the first time after booster immunization
The 22nd day after epidemic disease) start observation mouse arthritis incidence daily, record disease time and disease incidence, incidence=(morbidity
Mouse number of elements/total the number of elements of mouse) * 100.2) arthritis score: observation mouse arthroncus situation scores.Scoring is by 2
Experimenter is independent simultaneously to carry out, and the average value for taking two people to score is as final scoring.Scoring joint includes the front and back limb of mouse
Totally four podarthrums, each Joint scores standard are as follows: swelling toe is 1 point less than 2, and more than two is 2 points, sole swelling
It is 1 point, ankle swelling is 1 point.Each joint highest 4 is divided, and every mouse highest 16 is divided.The arthritis average mark of every group of mouse=
All mouse Joint scores summation/mouse numbers of elements.
(4) histopathology and scoring: to picture -- extremities joint；It is fixed -- the fixed 48h of 4% formalin；It is de-
Calcium -- decalcifying Fluid is handled 30 days；Slice -- longitudinal extending shaft is cut in half, and exposure marrow, marrow is face-down, and paraffin embedding is cut
Piece；Coverslip preservation is sealed up in dyeing -- conventional H E dyeing.Synovial tissue of joint's pathological score standard: 0 point: normal；1 point: synovial membrane
Hyperplasia and inflammatory cell infiltration；2 points: pannus is formed, articular cartilage damage；3 points: under most of articular cartilage damage and cartilage
Bone erosion；4 points: joint integrity is lost, arthrocleisis.As a result see Fig. 2.
Result of study
As can be drawn from Figure 2, in Primary preventive intervention model, at control group (CIA) mouse and high dose indolepopionic acid
The Arthritis Incidence of reason group (IPA=40mg/kg) is 90%, and low dosage indolepopionic acid group (IPA=20mg/kg) arthritis
Incidence be 50% (attached drawing 2a), it can be seen that only low dosage indolepopionic acid can significantly reduce the generation of experimental arthritis
Rate.Meanwhile the arthritis score average out to 8.87 of control group (CIA) mouse, low dosage indolepopionic acid group (IPA=20mg/kg)
Arthritic to be equally divided into 3.23, high dose group (IPA=40mg/kg) is 5.62 (attached drawing 2b), illustrates low dosage and high dose
Indolepopionic acid can significantly reduce arthritis score, inhibit Arthritis development.In addition, small to CIA using low dosage indolepopionic acid
Mouse carries out therapeutic intervention, and indolepopionic acid group mouse arthritis is equally divided into 8.78 as the result is shown, and control group mice is flat
11.83 (attached drawing 2c) are divided into, illustrates that the progress of CIA can be effectively suppressed in indolepopionic acid, alleviates the swelling symptom in mouse joint.With
Clinical score is consistent, the histopathology scoring average out to 1.89 in indolepopionic acid group mouse joint, and control group is then 2.51 (attached drawings
2d), the therapeutic effect of indolepopionic acid is further demonstrated.
(attached drawing 2a and attached drawing 2b) low dosage indolepopionic acid (IPA-20,20mg/kg) can significantly reduce experimental arthritis
Incidence arthritis score, and high dose indolepopionic acid (IPA-40,40mg/kg) to the disease incidence of experimental arthritis without bright
Development is rung, but can slightly reduce arthritis score.(attached drawing 2c and attached drawing 2d) indolepopionic acid (IPA, 20mg/kg) can significantly inhibit
The progress of experimental arthritis reduces arthritis score, and mitigates synovial tissue of joint's cell infiltration and cartilage destruction.
Embodiment 3
The ratio of indolepopionic acid regulation experimental arthritis mouse lymphocyte subgroup
Implementation method
Lymphocyte Subsets By Fcm ratio: (armpit is He popliteal nest drenches separation CIA mouse joint draining lymph node
Fawn on), lymph node is ground into single cell suspension using 1640 culture mediums and strainer by lymphonodi mesenterici and spleen, and adjustment is thin
Born of the same parents' quantity is 2-3 × 106/ 100 μ l, upper machine (BD after adding dye streaming antibody (eBioscience, U.S.A.), washing to be resuspended
FACSAriaTMII it) detects.As a result see Fig. 3.
Result of study
As can be drawn from Figure 3, lymphocyte subgroup analysis is the results show that in joint draining lymph node (DLN), indoles
Propionic acid lower t helper cell Th1 ratio (attached drawing 3a, CIA group Th1 2.1 ± 0.1755vs IPA group Th1 1.805 ±
0.1435), and the ratio of folliculus helper T lymphocyte Tfh and regulatory T cells Treg are without significant change (attached drawing 3b and c).In intestines
In mesentery lymph node (MLN), indolepopionic acid significantly lower Th1 cell ratio (attached drawing 3d, CIA group Th1 3.226 ±
0.5681vs IPA group Th1 1.375 ± 0.1559), the ratio of Tfh cell has down regulation trend (attached drawing 3e, CIA group Tfh
2.611 ± 0.3589vs IPA group Tfh 1.79 ± 0.328), and the ratio of Treg cell has up-regulation trend ((attached drawing 3f, CIA
Group 6.611 ± 0.7024vs of Treg IPA group Treg 8.218 ± 1.214).In spleen, indolepopionic acid lowers Th1 (attached drawing
3g, CIA group Th1 2.768 ± 0.3875vs IPA group Th1 1.906 ± 0.2403) and Tfh (attached drawing 3h, CIA group Tfh
1.743 ± 0.2114vs IPA group Tfh 1.154 ± 0.1673) cell ratio, the ratio of Treg cell has up-regulation trend (attached
Fig. 3 i, CIA group Treg 7.926 ± 0.9528vs IPA group Treg 9.29 ± 1.463).
Embodiment 4
The expression of indolepopionic acid regulation autoantibody and inflammatory mediator
Implementation method
ELISA kit detects cytokine-expressing: eyeball takes blood after mouse anesthesia, and whole blood stands 120min, and 4 DEG C,
1800rpm is centrifuged 20min.Collect upper serum, ELISA kit detect cell factor IL-6 and IL-10, operating method according to
According to kit (Multisciences Biotech Co., Ltd) explanation.As a result see Fig. 4.
Result of study
As can be drawn from Figure 4, the expression quantity of control group (CIA) mouse autoantibody anti-CII be 230974 ±
The expression quantity of 9296U/mL, low dosage indolepopionic acid group (IPA=20mg/kg) autoantibody anti-CII be 175314 ±
15123U/mL, and the expression quantity of high dose group (IPA=40mg/kg) is 204199 ± 34500U/mL, it can be seen that low dose
Amount indolepopionic acid can significantly lower the expression (attached drawing 4a) of autoantibody.Cytokines measurement is the results show that low indolepopionic acid can
The expression of proinflammatory cytokine IL-1 β, IL-6 and IL-17A are lowered, IL-1 β is in the content of CIA group and low dosage indole acid group point
Not Wei 12.66 ± 1.302pg/mL and 5.7 ± 0.411pg/mL, IL-6 the content of CIA group and low dosage indole acid group distinguish
For 194.3 ± 43.99pg/mL and 84.21 ± 16.64pg/mL, IL-17A is in the content of CIA group and low dosage indole acid group point
It Wei not 82.28 ± 16.47pg/mL and 50.48 ± 9.74pg/mL；The expression of other proinflammatory cytokines such as IFN-γ and TNF-α
Without significant change (attached drawing 4b-f).The expression of anti-inflammatory cytokines IL-10 and TGF-β becomes in low dosage indolepopionic acid group without obvious
Change, and TGF-β even has downward (attached drawing 4g and h) in high dose processing group.In addition, can to increase analgesia medium β-interior for indolepopionic acid
The expression (attached drawing 4i) of deltorphin delta (β-endorphin), the content of CIA group are 62.56 ± 1.294pg/mL, and indolepopionic acid group
Content is 68.18 ± 1.375pg/mL.
By above embodiments, it can be concluded that, indolepopionic acid can be by inhibiting proinflammatory cytokines Th1 and folliculus helper T lymphocyte
(Follicular helper T cell, Tfh) and raise anti-inflammatory cells regulatory T cells (T regulatory, Treg)
Ratio and play anti-inflammatory and immunosuppressive action, correct for the intracorporal immunocyte imbalance state of biology, and then reach prevention and treatment class
The purpose of rheumathritis.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (9)
1. application of the indolepopionic acid in preparation prevention and treatment rheumatoid arthritis drug.
2. application according to claim 1, which is characterized in that the drug further includes that indolepopionic acid is pharmaceutically subjected to
Auxiliary material.
3. indolepopionic acid lowers the application in histoorgan in the drug of the expression of proinflammatory cytokines Th1 in preparation.
4. indolepopionic acid reduces the application in histoorgan in the drug of the expression of Tfh cell in preparation.
5. application of the indolepopionic acid in preparation up-regulation histoorgan in the drug of the expression of Treg cell.
6. according to the described in any item applications of claim 3~5, which is characterized in that the histoorgan include joint lymph node,
Lymphonodi mesenterici or spleen.
7. indolepopionic acid reduces the application in serum in the drug of the expression of proinflammatory factor in preparation.
8. application according to claim 7, which is characterized in that the proinflammatory factor includes IL-1 β, IL-6 and IL-17A.
9. application of the indolepopionic acid in preparation up-regulation serum in the drug of the secretion of beta-endorphin.
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| CN113440522A (en) * | 2021-07-29 | 2021-09-28 | 中山大学附属第七医院(深圳) | Application of indolpropionic acid in preparation of drugs for treating autism |
| CN114831984A (en) * | 2022-06-20 | 2022-08-02 | 滨州医学院 | Application of indolpropanoic acid in preparation of drug for preventing and/or treating nephritis |
| WO2023066132A1 (en) * | 2021-10-19 | 2023-04-27 | 山东善维免疫科技有限公司 | Starch-indole acid derivative, preparation method therefor, and use thereof |
| CN117883439A (en) * | 2024-01-18 | 2024-04-16 | 中国人民解放军陆军军医大学第一附属医院 | Application of indole-3-propionic acid in preparation of medicines for inhibiting osteoclast differentiation |
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