CN104887656B - Application of the L-dopa methyl ester hydrochloride in the medicine for preparing preventing and treating rheumatoid arthritis - Google Patents
Application of the L-dopa methyl ester hydrochloride in the medicine for preparing preventing and treating rheumatoid arthritis Download PDFInfo
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Abstract
The present invention provides a kind of application of L-dopa methyl ester hydrochloride in the medicine for preparing preventing and treating rheumatoid arthritis, and L-dopa methyl ester hydrochloride chemical structural formula is as follows：.The L-dopa methyl ester hydrochloride that the present invention is provided is definite to rheumatoid arthritis, for L-dopa methyl ester hydrochloride exploitation is provided into strong foundation for new drug.
Description
Technical field
The invention belongs to technical field of new application of medicine, it is related to the medicinal usage of L-dopa methyl ester hydrochloride, and in particular to
Application of the L-dopa methyl ester hydrochloride in the medicine for preparing preventing and treating rheumatoid arthritis.
Background technology
Rheumatoid arthritis（Rheumatoid arthritis, RA）It is a kind of common with chronic, progressive, aggressivity
Systemic autoimmune disease with the characteristics of arthritis, it mainly invades the Minor articulus such as hand, wrist, foot, can also involve as the heart,
Other organ or tissues such as lung, nervous system.RA basic pathological changes are joint synovitis disease, pannus formation, articular cartilage
With the destruction of sclerotin progressive.Patient can have arthralgia, swelling, the destruction of stiff and permanent joint in the course of disease each stage, such as
Fail control in time, patient articular's deformity and function will eventually be caused to lose, with very high disability rate, trouble has been had a strong impact on
The quality of life of person.RA can fall ill in each age level, and its onset peak age concentrates on 30~50 years old, and general women suffers from
Person is more than male.According to statistics, disease incidence of disease in worldwide is about 0.5%~1.0%, and in China, illness rate is about
0.32%~0.36%.RA, which has become, causes crowd's disability and disabled one of the main reasons.
At present, RA therapeutic purposes are symptom managements, patient is reached remission state as early as possible, improve function of joint, most
Holding patient's body normal function of big degree.RA principle of reatment be early treatment, drug combination, therapeutic scheme individuation with
And functional training.Clinically treatment for RA be based on drug therapy, mainly including NSAIDs, improve the state of an illness
Antirheumatic drug, biological agent, glucocorticoid and autonomic drug preparation etc..Although the symptom of patient RA has been obtained substantially after treatment
Alleviation, prognosis is also significantly improved, but still suffer from the state of an illness easily repeatedly, adverse drug reaction, drug resistance etc. ask
Inscribe, therefore find new RA medicines tool and be of great significance.
L-dopa methyl ester hydrochloride（LDME）, chemical name is L-DOPA methyl esters hydrochloric acid, its point
Minor is C10H13NO4.HCL, and molecular weight is that 247.68, CAS No. are 1421-65-4, structure such as formula（Ⅰ）It is shown：
LDME is the pro-drug of levodopa, is hydrolyzed to after levodopa, enters maincenter by blood-brain barrier, through DOPA
Decarboxylation changes into dopamine, clinically for improving Parkinsonian symptoms.LDME is soluble in water, is easily made various oral
Formulation and injection administration, absorb fast, 7-8 minutes blood concentration peakings are more rapid-action than levodopa, curative effect affirmative.It is near several
Year animal experiment study shows that LDME also has therapeutic action to amblyopia.The pass of research LDME and rheumatoid arthritis is there is no at present
System, the present invention comes therefrom.
The content of the invention
The technical problem of solution：For existing research LDME and the shortage of the relation of rheumatoid arthritis, the present invention
A kind of application of L-dopa methyl ester hydrochloride in the medicine for preparing preventing and treating rheumatoid arthritis is provided, for preventing and treating rheumatoid arthrosis
Inflammation provides a kind of new way.
Technical scheme：Application of the L-dopa methyl ester hydrochloride in the medicine for preparing preventing and treating rheumatoid arthritis, hydrochloric acid is left
Revolve dopa methyl ester chemical structural formula as follows：
。
Described pharmaceutical composition includes the L-dopa methyl ester hydrochloride of pharmaceutical effective dose and pharmaceutically acceptable
Carrier.
The described pharmaceutical preparation being prepared into L-dopa methyl ester hydrochloride suitable for intestines and stomach or parenteral administration.
The described pharmaceutical preparation being prepared into L-dopa methyl ester hydrochloride suitable for intestines and stomach, its dosage form is conventional
Tablet, capsule, controlled release preparation or sustained release preparation.
It is described in the pharmaceutical preparation that L-dopa methyl ester hydrochloride is prepared into, L-dopa methyl ester hydrochloride is in the formulation
Content is 1wt%~99wt%.
It is described in the pharmaceutical preparation that L-dopa methyl ester hydrochloride is prepared into, L-dopa methyl ester hydrochloride is in the formulation
Content is 10wt%~90wt%.
The preparation method of described pharmaceutical preparation is that L-dopa methyl ester hydrochloride directly made into preparation, or respectively or/
Preparation is made with after auxiliary material mixing, packaging is then carried out and produces.
The dosage of described pharmaceutical composition enters according to the dosage form difference of administration object, method of administration or medicine
Row change, but premised on ensureing that the pharmaceutical composition can reach effective blood concentration in mammalian body.
L-dopa methyl ester hydrochloride（LDME）, its molecular formula is C10H13NO4.HCL, chemical name is L-3,4- dihydroxy benzenes
Base methyl lactamine hydrochloric acid, molecular weight is that 247.68, CAS No. are 1421-65-4, from Sigma-Aldrich
Co.LLC. company.
Can technical solution of the present invention studies LDME by intraperitoneal injection method set up class wind to collagen-induced method
Wet arthritic mice（CIA）There is therapeutic action, while determining tumor necrosis factor-alpha（TNF-α）, Interleukin -1β（IL-1β）With
The inflammatory Cytokines Expressions such as IL-6 change, to provide experimental basis to inquire into LDME treatment rheumatoid arthritis.
The present invention is randomly divided into 5 groups, i.e. normal group by 40 DBA/1j mouse（Control）, model group
（Vehicle）, positive controls（Methotrexate (MTX), 2mg/kg；MTX）, LDME high doses group and LDME low dose groups, the high agent of LDME
The dosage of amount group is 100mg/kg, and the dosage of LDME low dose groups is 25mg/kg.In secondary immunity the previous day（The
20 days）100mg/kg and 25mg/kg successive administrations are pressed respectively to the 70th day LDME group, and MTX groups are administered for every 2 days by 2mg/kg,
Control and Vehicle groups give drinking water by 10ml/kg.Routine observation each group experiment mice ordinary circumstance, record mouse foot
Slap thickness, arthritis score；Each group put to death animal in the 70th day and retains serum and hindlimb joints sample, respectively row iconography, group
Knit and molecular Biological Detection.Statistical analysis is carried out to each group of data using one-way analysis of variance.
Result of the present invention shows that flexibly, the state of mind is good for the activity of Control groups mouse, and drinking-water and appetite are normal, joint
Activity is freely.Vehicle group mouse are One's spirits are drooping, and drinking-water and appetite are reduced, and joint motion has obvious obstacle, and extremities joint is obvious
Enlargement, lasting oedema, dermohemia, skin temperature rise, range of motion are small.The LDME low dose group mouse state of mind is general, drink
Water and the situation of ingesting are better than Vehicle groups, and joint motion is limited, and partial joint is red and swollen, and locomotor activity obtains part recovery.LDME
High dose group and MTX group mouse diets situation are significantly better than Vehicle groups, and preferably, joint is slightly red and swollen for the state of mind, without substantially
Moving obstacle.Start gradually swelling within the 21st day after Vehicle group mouse vola initial immunities, to swelling degree at the 42nd day up to most
Height, is compared, difference is statistically significant with Control groups（p<0.05）.LDME low dose groups, mouse vola swelling enters to postpone
Slowly, initial immunity is after the 42nd day, and mouse vola swelling degree gradually mitigates, last measure when, mouse vola swell thickness with
Vehicle groups compare, and difference is statistically significant（p<0.05）.Vehicle groups arthritis score peaked in the 42nd day；
MTX groups were significantly reduced in D38 days arthritis scores；LDME high doses group and LDME low dose groups respectively the 36th, the 42nd day it is aobvious
Write and suppress CIA mouse arthropathies.Micro-CT scanning results show, LDME low dose groups：Bony surface is very crude, joint
Space loss is serious, and articulation structure destruction is more serious, is slightly taken a turn for the better compared with Vehicle groups；LDME high doses group and MTX groups：Sclerotin
Surface is substantially complete, and joint space is almost normal, and articulation structure is high-visible.Histological stain result shows that Control groups are closed
Save cartilage queueing discipline, even dyeing；Articular surface is complete, and cartilage is in pink light dyeing, and sclerotin is in peony, and joint space is just
Often, synovial tissue's structure is smooth, has no synovial hyperplasia and inflammatory cell infiltration phenomenon.Vehicle groups are mainly shown as：Synovial membrane group
Notable hyperplasia is knitted, a large amount of inflammatory cell infiltrations, new vessels showed increased occurs；Cartilage cell's arrangement disorder, articular cartilage and
Sclerotin is seriously damaged, and cartilage surface is crude uneven, and joint space disappears.LDME high doses group, LDME low dose groups and MTX
Treatment group can mitigate above pathological change in various degree, show as the slight hyperplasia of synovial tissue, and inflammatory cell infiltration is reduced, newborn
Blood vessel is reduced, and cartilage damage degree substantially mitigates.ELISA results show that LDME can reduce TNF-α in CIA mice serums, IL-1 β
With IL-6 content, prompting is probably the important mechanisms of LDME treatment rheumatoid arthritis to the adjustment effect of inflammatory factor.
The zoopery confirms L-dopa methyl ester hydrochloride（LDME）There is certain preventive and therapeutic effect to rheumatoid arthritis, can
Destruction of joint and inflammatory reaction is inhibited, can as the pharmaceutical intervention of rheumatoid arthritis a kind of new tool.
Beneficial effect：The L-dopa methyl ester hydrochloride that the present invention is provided is in the medicine for preparing preventing and treating rheumatoid arthritis
Using with advantages below：
1. L-dopa methyl ester hydrochloride has the purposes of efficiently preventing and treating rheumatoid arthritis, it can significantly mitigate symptom, such as
Arthroncus, arthritis score, and destruction of joint can be mitigated, reduce Secretion of Inflammatory Factors；
2. L-dopa methyl ester hydrochloride is definite to rheumatoid arthritis, to be by L-dopa methyl ester hydrochloride exploitation
New drug provides strong foundation.
3. the present invention has excavated the new prospect in medicine of L-dopa methyl ester hydrochloride, a new medicinal applications have been opened up；Salt
Sour L-dopa methyl ester is clinically to be used to improve the common drug of Parkinsonian symptoms, and convenient to take, toxic side effect is small, for a long time
Medication non-hazardous.
Brief description of the drawings
Fig. 1 is mouse hind leg foot in Control groups, Vehicle groups, MTX groups, LDME high doses group, LDME low dose groups
Slap thickness change figure.
Fig. 2 is mouse arthritis in Control groups, Vehicle groups, MTX groups, LDME high doses group, LDME low dose groups
Scoring figure.
Fig. 3 is mouse micro- in Control groups, Vehicle groups, MTX groups, LDME high doses group, LDME low dose groups
CT testing result figures.
Fig. 4 is mouse knee joint in Control groups, Vehicle groups, MTX groups, LDME high doses group, LDME low dose groups
Histo pathological change figure.
Fig. 5 is mice serum in Control groups, Vehicle groups, MTX groups, LDME high doses group, LDME low dose groups
ELISA testing result figures.
Embodiment
Embodiment 1
First, materials and methods
1. material
1.1 reagents and experimental facilities
1.1.1 primary drug and reagent
L-dopa methyl ester hydrochloride（LDME）, dimethyl sulfoxide（DMSO）, hydrogen peroxide, the water of disodium EDTA two
Compound（EDTA）Purchased from Sigma, the U.S.；The Collagen Type VI of ox II, incomplete Freund's adjuvant（IFA）, complete Freund's adjuvant（CFA）It is purchased from
Chondrex, the U.S.；Methanol, hydrochloric acid, glacial acetic acid, ammoniacal liquor, absolute ethyl alcohol, 95% ethanol, paraformaldehyde are tried purchased from Shanghai peaking
Agent Co., Ltd；ColeShi haematoxylin dyeing liquid（Normal dyeing）, eosin stains liquid be purchased from Beijing Suo Laibao Science and Technology Ltd；
TNF-α, IL-1 β, IL-6 enzyme-linked immunosorbent adsorption test detection kits, purchased from Biosource, the U.S..
1.1.2 key instrument
Micro-CT（SkyScan 1176, Belgium）, paraffin slicing machine（Leica 2135, Germany）, roasting piece machine
（Leica 1120, Germany）, paraffin wax embedding（BMJ- II, China, Changzhou）, Axio imager.MI just put microscope
（Zeiss, Germany）, ELIASA（Biotec, the U.S.）, Sim-F140 ice machines（Sanyo, Japan）, operating theater instruments it is a set of etc..
1.2 experimental animal
Male DBA/1j mouse, 40, body weight 20-25g, 6-8 week old is limited purchased from the magnificent Fukang biotechnology share in Beijing
Company.Experimental animal is raised in University Of Suzhou SPF grades of experiment room, room temperature（23±1）DEG C, relative humidity（55±10）%,
Interval illumination, is freely ingested and drinks water, and adaptability is used after raising one week.
2. experimental method
The foundation and evaluation of 2.1 mouse collagen Induced Arthritis models
2.1.1 the foundation of mouse collagen Induced Arthritis model
（1）The preparation of emulsifying agent：By a certain amount of ox II types（CII）Collagen is dissolved in 10 mM glacial acetic acid, is configured to dense
The solution for 2mg/mL is spent, 4 DEG C are stirred overnight.By the collagen solution being completely dissolved and not formula adjuvant completely（Incomplete Fu Shizuo
Agent）Isometric mixing, the two is mixed, be allowed to fully emulsified completely using three-way pipe, and it is lmg/mL's to be made containing collagen concentration
Emulsion.Emulsifying agent need it is now with the current, and should be positioned in ice face to prevent temperature rise cause collagenous degeneration.
（2）Mouse is put into fixator and exposes tail, left hand gently pinches mouse tail root, the right hand holds micro syringe
The inserting needle at away from root of the tail portion on the lower 1.5-2.0cm, the μ L emulsifying agents of intracutaneous injection 100.If there is emulsifying agent to overflow in bolus infusion processes
Go out, then new injection point should be selected to carry out the appropriate emulsifying agent of supplement injection.
（3）First time inoculation takes the emulsifying agent booster shots of same dose once after 3 weeks.Note：This emulsifying agent
Preparation is mixed using freurd incomplete adjuvant with collagen.
（4）Different degrees of redness occurred in 4-5 days mouse after second of booster immunization, to initial immunity 38 days or so
Peak, general 45 days or so up to more than 95% into mould rate.
2.1.2 experiment packet and administration
All experimental animals are divided into 5 groups using random digits table（Every group 8）, i.e. normal group（Control）, model
Group（Vehicle）, positive controls（Methotrexate (MTX), 2mg/kg；MTX）, LDME components for height（H-LDME）With it is low（L-LDME）
Two dosage groups, its dosage is respectively 100mg/kg and 25mg/kg.In secondary immunity the previous day（20th day）To the 70th
Its LDME group presses 100mg/kg and 25mg/kg successive administrations respectively, and by 2mg/kg administrations, Vehicle groups were trans-abdominal in every 2 days for MTX groups
Chamber injecting normal saline 0.1ml/kg/d, Control group mouse conventinal breeding, is disregarded.
2.1.3 the evaluation of mouse collagen Induced Arthritis model
（1）Mouse metapedes vola thickness measure：From D21, metapedes vola thickness of the slide measure to mouse homonymy is used
Measure, measure 2 times weekly, each duplicate measurements 3 times is averaged.All measurements are by two experimenter's complete independentlies.
（2）Arthritis and the scoring of swelling degree：From D21, each group rat articular swelling degree is scored, weekly
Evaluation and test 2 times, each duplicate measurements 3 times, averages.Each arthropathy degree is evaluated by 5 grades of point systems, and each limbs are most
High score is 4 points, and every mouse is up to 16 points.Specific standards of grading are as follows：0 point：Without redness；1 point：Small toe joint is slightly red
It is swollen；2 points：Toe joint and vola pedis arthroncus；3 points：Sufficient pawl swelling below ankle-joint；4 grades：Whole including ankle-joint
There is swelling in sufficient pawl, and function completely loses.
2.3 collection of specimens
Mouse orbit veniplex is taken a blood sample, and mouse puts to death preceding with orbital venous plexus blood collection method blood sampling 0.25ml, and specific method is such as
Under：The capillary glass tube that internal diameter is 1.0-1.5mm is taken, with the capillary for fractureing into 1-1.5cm length before, the leaching of 1% heparin solution
Bubble, is used after drying.After left hand fixes mouse, neck both sides are oppressed downwards, and the right hand holds capillary blood sampling tube, accesses ready
In container.Gauze gently presses hemostasis after blood sampling.Sample centrifuges 10min with 3000g, obtains -70 DEG C of preservations of serum.
Mouse joint sample：10% chloral hydrate anesthesia is injected intraperitoneally in each group animal, lies on the back and is fixed on mouse crosshead outside
On, chest exposure heart is opened, heart catherization is irrigated to the left through the apex of the heart, cut off open physiological saline after right auricle of heart, ligation descending aorta
Rinse to right auricle of heart and flow out refrigerant liquid, then irrigated with 4% 200~300ml of neutral paraformaldehyde, to animal foot tic, become
Firmly.Perfusion is rapid after finishing to take out bilateral hind leg, rejects basis cranii attaching soft tissue, retains knee joint and toes joint, -70 DEG C of guarantors
Deposit.
2.4 Micro-CT are analyzed
Put to death mouse within the 70th day after initial immunity, take out mouse rear solid end, 10% paraformaldehyde is fixed after 48 hours using micro-
CT （SkyScan 1076）All samples are scanned, the parameters of scanning set as follows：9 μm of spatial resolution, voltage
70 kV, electric current 141 μ A, sweep time 1750ms, after the completion of scanning, are analyzed mouse ankle joint using the software carried
And build the three-dimensional reconstruction image of sample.
2.5 histopathology histologies
2.5.1 mouse joint pathology microsection manufacture
（1）Fixed, decalcification：D70 puts to death mouse, takes out rear solid end, 48 hours are fixed with 4% paraformaldehyde, 10% is subsequently placed in
Decalcification 1 month in EDTA decalcifying Fluids, 2d changes liquid 1 time,
（2）Dehydration, waxdip：Sample after decalcification is rinsed into 30-60min using flowing water, is then dehydrated, waxdip, had
Body flow is shown in Table：
（3）Embedding：Embedded on embedding machine, mouse knee joint section is put down and presses on embedded box bottom and is adjacent to, stone is treated
Wax stone is released after wax solidification, solid wax stone is made.
（4）Section, exhibition piece, roasting piece：The wax stone trimmed is cut into slices, sample is cut into thickness with rotary microtome is
4-5 μm of serial section, water-bath exhibition piece in exhibition piece machine is put into front（43 DEG C of water temperature）, it is as far as possible smooth, non-wrinkled during exhibition piece,
With clean slide list piece, drag for section to be placed on roasting piece machine after the completion of piece and carry out roasting piece（Temperature 60 C）4 hours, prevent gas
Bubble is produced.
2.5.2 Hematoxylin-eosin is dyed（H＆E is dyed）
H＆E decoration methods, are the most frequently used colouring methods of morphology, while being also the decoration method commonly used in paraffin section technology
One of.The morphosis of various tissues or cell is observed that by this colouring method.
Experimental procedure：
（1）60 DEG C of baking ovens bake piece 30min.
（2）Paraffin section is placed in dimethylbenzene I, II, each 15min.
（3）Absolute ethyl alcohol I, II, 95% ethanol I, II, each 5 minutes of 90% ethanol, 80% ethanol, 70% ethanol.
（4）Haematoxylin dyeing 1-3min, running water rinses 1min.
（5）1% acidic alcohol breaks up 20 seconds.
（6）1% weak aqua ammonia returns blue 30 seconds, running water flushing 1min.
（7）Eosin stains 3 minutes, originally wash 5min.
（8）It is conventional dehydration, transparent（80% ethanol 5min, 90% ethanol 5min, 95% ethanol 5min, absolute ethyl alcohol 5min,
Transparent I 10min of dimethylbenzene, transparent II 10min of dimethylbenzene）.
（9）Neutral gum mounting.
（10）Micro- Microscopic observation Color：Nucleus is in blueness, and cytoplasm, connective tissue etc. takes on a red color.
2.6 enzyme-linked immunosorbent assay（ELISA）Detect TNF-α, IL-1 β and IL-6
TNF-α, IL-1 β and IL-6 in ELISA detection serum.
With reference to ELISA kit specification, comprise the following steps that：
（1）The hole of gauge orifice 8 is set up, the μ l of sample diluting liquid 100 are added per Kong Zhongxian, the first hole adds the μ of standard items 100 again
L, suctions out 100 μ l with pipettor after mixing, the second hole is moved to, so oppose repeatedly is diluted to the 7th hole again, finally, from the 7th hole
100 μ l of middle sucking-off are discarded, and it is 100 μ l to be allowed to volume.8th hole is blank control.
（2）Sample-adding:The μ l of supernatant 100 are added per hole in testing sample hole, reaction plate is placed in 37 DEG C × 120
min；
（3）Reaction plate is fully washed 4~6 times with cleaning solution, to filter paper on buckle dry, per hole in add first antibody work
Liquid 50ul, reaction plate is fully mixed rearmounted 37 DEG C × 60 min.
（4）Reaction plate is fully washed 4~6 times with cleaning solution, to filter paper on buckle dry, per the enzyme-added labeling antibody working solution in hole
100ul, 37 DEG C of 120 min is placed in by reaction plate；
（5）Reaction plate is fully washed 4~6 times with cleaning solution, to filter paper on buckle dry, per hole in add substrate working solution
100ul, is placed in 37 DEG C of dark places and reacts 5 ~ 10 min；
（6）50ul terminate liquids are added in per hole to mix, and light absorption value is surveyed at 450nM with ELIASA；
（7）With the pg/ml of standard items 1000,500,250,125,62.5,31.25,15.625,0 OD values in semilog
Mapped on paper, draw standard curve；Testing index content in serum is conversed according to calibration curve formula.
2.7 statistical analysis
Result data is analyzed using SPSS11.0 statistical softwares, data mean ± standard deviation（）Represent, multigroup ratio
Relatively select one-way analysis of variance（One-way ANOVA are examined）, compare two-by-two under conditions of population variance is neat, from LSD
And Dunnett-t methods are analyzed.p<0.05 is that difference is statistically significant.
2nd, result
1. experimental animal ordinary circumstance
Without dead mouse in experimentation.Flexibly, the state of mind is good for the activity of Control groups mouse, and drinking-water and appetite are just
Often, joint motion is freely.Vehicle group mouse are One's spirits are drooping, and drinking-water and appetite are reduced, and joint motion has obvious obstacle, four limbs
The obvious enlargement in joint, lasting oedema, dermohemia, skin temperature rise, range of motion are small.The L-LDME group mouse state of mind one
As, drink water and the situation of ingesting is better than Vehicle groups, joint motion is limited, and partial joint is red and swollen, and it is extensive that locomotor activity obtains part
It is multiple.H-LDME groups and MTX group mouse diets situation are significantly better than Vehicle groups, and preferably, joint is slightly red and swollen, nothing for the state of mind
Obvious moving obstacle.
2. influences of the LDME to CIA mouse vola thickness
Since after initial immunity the 21st day, the measurement of vola thickness is carried out to each group right side of mice metapedes, 2 are measured weekly
It is secondary.Start gradually swelling within the 21st day after Vehicle group mouse vola initial immunities, highest is reached to swelling degree at the 42nd day, with
Control groups compare, and difference is statistically significant（p<0.05）.LDME treatment groups, mouse vola swelling is made slow progress, and is exempted from for the first time
After epidemic disease the 42nd day, mouse vola swelling degree gradually mitigates, when last is measured, and the swollen thickness of mouse vola is compared with Vehicle groups,
Difference is statistically significant（p<0.05）.See Fig. 1.
3. the influence that LDME scores CIA mouse arthritis
There is red and swollen, deformation in hindlimb joints after Vehicle group mouse secondary immunities, and arthritis score reached height in D42 days
Peak；MTX groups were significantly reduced in D38 days arthritis scores；LDME（High dose and low dosage）Respectively in D36, D42 is significantly inhibited
CIA mouse arthropathies, are compared, difference is statistically significant with Vehicle groups（p<0.05）.See Fig. 2.
4. influences of the LDME to CIA mouse destruction of joint
Micro-CT scanning results show, Control groups：Bony surface is smooth, and joint space is normal, and articulation structure is complete
Whole, sclerotin does not occur breakoff phenomenon；Vehicle groups：Bony surface is uneven, and articulation structure destruction is serious, and joint space is complete
It totally disappeared mistake；L-LDME groups：Bony surface is very crude, and joint space constriction is serious, and articulation structure destruction is more serious, compared with model group
Slightly take a turn for the better；H-LDME and MTX groups：Bony surface is substantially complete, and joint space is almost normal, and articulation structure is high-visible.See
Fig. 3.
5. each group mouse joint tissue pathological change
Control group articular cartilage queueing disciplines, even dyeing；Articular surface is complete, and cartilage is in pink light dyeing, and sclerotin is in
Peony, joint space is normal, and synovial tissue's structure is smooth, has no synovial hyperplasia and inflammatory cell infiltration phenomenon.Vehicle groups
It is mainly shown as：, there are a large amount of inflammatory cell infiltrations, new vessels showed increased in the notable hyperplasia of synovial tissue；Cartilage cell arranges
Row are disorderly, and articular cartilage and sclerotin are seriously damaged, and cartilage surface is crude uneven, and joint space disappears.LDME and MTX treatments
Group can mitigate above pathological change in various degree, show as the slight hyperplasia of synovial tissue, and inflammatory cell infiltration is reduced, new vessels
Reduce, cartilage damage degree substantially mitigates.See Fig. 4.
6. influences of the LDME to CIA mice serums TNF-α, IL-1 β and IL-6
Vehicle group mice serums TNF-α, IL-1 β and IL-6 content are passed through apparently higher than normal group by table is visible
After LDME treatments, H-LDME group IL-1 β and IL-6 content are substantially reduced, and TNF-α is significantly reduced in each treatment group, with
Vehicle groups compare, and difference is statistically significant（p<0.05；See Fig. 5）.The above results point out the adjustment effect to inflammatory factor
It is probably the important mechanisms that LDME treats rheumatoid arthritis.
Conclusion：LDME can significantly reduce TNF-α in CIA mice serums, IL-1 β and IL-6 content, hence it is evident that suppress CIA
Mouse arthritis disease, and the destruction of CIA mouse inflamed joint can be substantially reduced.LDME has obvious curative effect to CIA mouse, is clinical
The drug therapy of rheumatoid arthritis provides new selection.
Claims (5)
1. application of the L-dopa methyl ester hydrochloride in the medicine for preparing preventing and treating rheumatoid arthritis, L-dopa methyl ester hydrochloride
Learn structural formula as follows：
δ????-1。
2. L-dopa methyl ester hydrochloride according to claim 1 answering in the medicine for preparing preventing and treating rheumatoid arthritis
With, it is characterised in that L-dopa methyl ester hydrochloride is prepared into the pharmaceutical preparation suitable for intestines and stomach or parenteral administration.
3. L-dopa methyl ester hydrochloride according to claim 1 answering in the medicine for preparing preventing and treating rheumatoid arthritis
With, it is characterised in that L-dopa methyl ester hydrochloride is prepared into the pharmaceutical preparation suitable for intestines and stomach, its dosage form is conventional
Tablet, capsule, controlled release preparation or sustained release preparation.
4. L-dopa methyl ester hydrochloride according to claim 3 answering in the medicine for preparing preventing and treating rheumatoid arthritis
With, it is characterised in that in the pharmaceutical preparation that L-dopa methyl ester hydrochloride is prepared into, L-dopa methyl ester hydrochloride is in the formulation
Content is 1wt%~99wt%.
5. L-dopa methyl ester hydrochloride according to claim 4 answering in the medicine for preparing preventing and treating rheumatoid arthritis
With, it is characterised in that in the pharmaceutical preparation that L-dopa methyl ester hydrochloride is prepared into, L-dopa methyl ester hydrochloride is in the formulation
Content is 10wt%~90wt%.
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