CN109793751A - Application of the araboxylan sulfonate in the drug of preparation treatment osteoarthritis - Google Patents
Application of the araboxylan sulfonate in the drug of preparation treatment osteoarthritis Download PDFInfo
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Abstract
The invention discloses a kind of application of araboxylan sulfonate in the drug of preparation treatment osteoarthritis, belong to field of pharmaceutical technology.Present inventor is by a large amount of research discovery, araboxylan sulfonate has obviously arthritis treatment effect, it can not only diminish inflammation, it can also promote repair of cartilage, it to a certain extent can be with reverse disease, therefore, araboxylan sulfonate can be used for preparing the drug for the treatment of osteoarthritis.
Description
Technical field
The present invention relates to a kind of application of araboxylan sulfonate in the drug of preparation treatment osteoarthritis, belong to
Field of pharmaceutical technology.
Background technique
Osteoarthritis (Osteoarthritis, OA) is the chronic degenerative disease of articular cartilage, and clinical manifestation is
Arthralgia, stiff, swelling and the limitation of activity slowly developed.The illness rate of China 60 years old or more crowd OA has been up to
50%, and then reach within 75 years old or more 80%, the disease have become seriously affect life of elderly person quality most common disease it
One.
Currently, mainly having three kinds of oral drugs, joint cavity injection drug and operative treatment schemes to the treatment of OA.Wherein,
Joint cavity injection belongs to centesis, to high operation requirements and is not easy repeated multiple times use, mainly in, the symptom of severe patient
Alleviate and improve the state of an illness.Operative treatment then mainly for serious middle and advanced stage patient, can thoroughly be eliminated using joint replacement
Pain.In clinic using it is most be oral drugs, including antalgesic and cartilage protection medicine two major classes, usual two class are medication combined
It uses, while achieving the purpose that analgesia and promoting repair of cartilage.Antalgesic mainly includes nonsteroidal anti-inflammatory drug, such as Diclofenac
Sodium, Meloxicam, celecoxib etc.；Analgesic-antipyretic Paracetamol, aspirin etc.；Opium analgesics such as codeine,
Tramadol hydrochloride etc..The most commonly used is Glucosamine, chondroitin sulfate, diacereins etc. for cartilage protection medicine.In addition, the above town
The certain external preparations and external Chinese medicament of pain medicine clinically also have application.But drug therapy can only be right whithin a period of time
The arthritis of patient improves, and there is no drug that the state of an illness can be reversed at present, thoroughly cures.
Largely there is acid mucopolysaccharide, including chondroitin sulfate, hyaluronic acid etc. in joint of animal cell, they are in body
Inside play important physiological function, such as lubricating joint, adjusting Vascular Permeability, regulatory protein matter, preventing adhesions etc..Cause
This, chondroitin sulfate, hyaluronic acid itself are also used for the reparation and rehabilitation of articular cartilage damage, so passing through sulphation
Or the means of sulfonation prepare related polysaccharides derivative and become to find the important directions of arthritis treatment drug.
Araboxylan is the New Dietary Fiber ratified by FDA in 2018, is concentrated mainly at present to its research
In the physiological action of itself, and it is very few to the research of its derivative.Currently, there has been no exist about araboxylan sulfonate
The relevant report of application in preparation treatment osteoarthritis drugs.
Summary of the invention
The purpose of the present invention is overcome the deficiencies of the prior art and provide a kind of araboxylan sulfonate to treat in preparation
Application in the drug of osteoarthritis.Present inventor is by a large amount of the study found that araboxylan sulfonate has
There is obviously arthritis treatment effect, can not only diminish inflammation, repair of cartilage can also be promoted, it to a certain extent can be with
Reverse disease, therefore, araboxylan sulfonate can be used for preparing the drug for the treatment of osteoarthritis.
The technical scheme to solve the above technical problems is that araboxylan sulfonate is closed in preparation treatment bone
Save the application in scorching drug.
Present inventor after study, obtains as drawn a conclusion:
First point: araboxylan sulfonate is made by diminishing inflammation and promoting cartilage nutrition to play osteoarthritis treatment
With significant in efficacy to osteoarthritis rabbit, effect is significantly better than commercial oral pharmaceutical composition (celecoxib and sulfuric acid amino
Glucose).
Second point, the drug effect of araboxylan sulfonate drug administration by injection is better than the drug effect being administered orally.Inventor's supposition,
Oral administration is likely due to macromolecular structure and is unfavorable for alimentary canal absorption or is possible to by the various enzymes of alimentary canal and enteron aisle
The decomposition of microorganism cannot reach affected part in the form of effective component, therefore drug administration by injection is better choice.
Thirdly, the structure of araboxylan sulfonate itself has apparent influence to curative effect, wherein branched structure and
The drug effect of the araboxylan sulfonate of the high raw material preparation of arabinose content is more preferable.
Based on the above technical solution, the present invention can also be improved as follows.
Further, the general structure of the araboxylan sulfonate are as follows:
In formula, the summation of x, y and z are 1-19；Side chain sulfonation arabinose passes through 1- oxygen glycosidic bonds and main chain sulfonation xylose
2- or/and 3- connection, and the molar ratio of the two be 1:2-1:20；Side chain miscellaneous sugar passes through 1- oxygen glycosidic bonds and main chain sulfonation wood
The 2- of sugar or/and 3- connections；Miscellaneous sugar in the sulfonation miscellaneous sugar be glucose, glucuronic acid, 4- methyl glucose uronic acid,
One of galactolipin, rhamnose, galacturonic acid, xylose are a variety of.
Be using above-mentioned further beneficial effect: above-mentioned araboxylan sulfonate is passed through by araboxylan
It is obtained after sulfonating reaction.The quantity of side chain sulfonation arabinose on araboxylan sulfonate high polymer main chain increases,
Be conducive to be promoted the curative effect of araboxylan sulfonic acid salts for treating osteoarthritis.
Further, the preparation method of the araboxylan sulfonate, includes the following steps:
Step 1: by araboxylan and sulfonating agent 1:(1-5 in molar ratio), being placed in reaction kettle together, obtain first
Mixture；
Step 2: the ratio of 100mL polar organic solvent is added according to araboxylan described in every (1-8) g step 1, to
Polar organic solvent is added in the first mixture that step 1 obtains, warming while stirring is to 55 DEG C -95 DEG C, after reaction 1-3 hours
It is down to room temperature, obtains the second mixture；
Step 3: lye being added into the second mixture that step 2 obtains, sulfonation described in the alkali in the lye and step 1
The molar ratio of agent is 1:1, and after being sufficiently stirred, obtained reaction mixture is added to 2-5 times of polar organic solvent described in step 2
In the ethyl alcohol of volume, sufficiently rear filtering to be precipitated obtains filter residue；
Step 4: according to every 10g filter residue be dissolved in (50-100) mL water ratio carry out it is water-soluble, with alkali identical in step 3
It is 7 that liquid, which adjusts pH value, and the ethyl alcohol that 2-5 times of volume of above-mentioned water is added carries out alcohol precipitation, and sediment is filtered, and repeats above-mentioned water-soluble, alcohol
Heavy, filtering operation twice, is collected last sediment, is dissolved in deionized water, and ultrafiltration membrane is crossed, and gained trapped fluid is chilled dry
It is dry to get arrive the araboxylan sulfonate.
It is using above-mentioned further beneficial effect: in polar organic solvent, is come with araboxylan and sulfonating agent
Prepare araboxylan sulfonate, sulfonation is high-efficient, also avoid in the prior art pyridine as serious caused by solvent
Environmental pollution burden.
Further, the sulfonating agent is sulfur trioxide trimethylamine compound, sulfur trioxide triethylamine compound, three oxidations
One of thiopyridines compound or sulfur trioxide N,N-dimethylformamide compound；The polar organic solvent is dimethyl
One of sulfoxide, N,N-dimethylformamide or DMAC N,N' dimethyl acetamide；The room temperature is 20-25 DEG C；The lye is
NaOH solution, KOH solution, Ca (OH)2One of solution；The molecule interception of the ultrafiltration membrane is 600Da.
Further, the araboxylan is extracted from monocotyledon.
Be using above-mentioned further beneficial effect: araboxylan is primarily present in monocotyledon hemicellulose
In xylan, the corresponding xylan being primarily present in dicotyledon hemicellulose.Two kinds of xylans remove
The plant in source is significantly different outer, and there is also huge difference for the chemical structure of itself, maximum difference is dicotyledon
Xylan is entirely free of branch or the even position 2- for having very small amount of glucuronic acid branch to be connected to xylose based on single-stranded
On.And monocotyledon araboxylan is branched structure, containing a large amount of branches, and branch is existed simultaneously in main chain xylose
2- and 3- hydroxyls on, the principal mode of branch is arabinose simultaneously with a small amount of other miscellaneous sugars.Present inventor
By largely testing discovery, although the xylan of two class plant origins all has certain treatment model after sulfonating reaction
The ability of rabbit osteoarthritis, but it is more significant to be derived from monocotyledonous araboxylan sulfonate curative effect, structure
In arabinose branching content have positive correlation effect to whole effect.By more different plant origins, structure and Ah
The sulfonate for the xylan preparation for drawing primary sugared content different, it can be found that the list that branch is abundanter, arabinose content is higher
Leaf plant araboxylan sulfonate is better to the therapeutic effect of rabbit osteoarthritis.It therefore, should be from monocotyledonous
The very potential therapeutic agent as osteoarthritis of araboxylan sulfonic acid.
Further, the monocotyledon be one of sugarcane, bamboo, corn, oat, reed, rice, wheat or
It is a variety of.
It is using above-mentioned further beneficial effect: extracts araboxylan from above-mentioned monocotyledon, more favorably
In industrialized production.
Further, the drug is using a effective amount of araboxylan sulfonate as active constituent, in addition pharmaceutically may be used
The pharmaceutical preparation that the carrier and/or auxiliary material of receiving are prepared.
It is using above-mentioned further beneficial effect: improves the mouldability of araboxylan sulfonate, validity and steady
It is qualitative.
Further, the dosage form of the drug is injection.
Be using above-mentioned further beneficial effect: compared to other dosage forms, injection can allow drug to be directly entered
Blood circulation reaches affected part, and the degradation of araboxylan sulfonate drug will not be caused because of alimentary canal.
Further, the injection includes aqueous solution or water composite solution.
Further, the administration mode of the drug is one of subcutaneous injection, intramuscular injection, intravenous injection or a variety of.
Be using above-mentioned further beneficial effect: drug administration by injection can allow drug to be directly entered blood circulation and reach trouble
Place, will not cause the degradation of araboxylan sulfonate drug because of alimentary canal.
The beneficial effects of the present invention are:
1. present inventor is by a large amount of the study found that araboxylan sulfonate has obviously closes
Save scorching therapeutic effect, can not only diminish inflammation, repair of cartilage can also be promoted, to a certain extent can with reverse disease, therefore,
Araboxylan sulfonate can be used for preparing the drug for the treatment of osteoarthritis.
2. the inventors of the present application found that medicine of the drug effect of araboxylan sulfonate drug administration by injection better than oral administration
Effect.
3. the inventors of the present application found that branched structure and arabinose content it is high raw material preparation araboxylan
The drug effect of sulfonate is more preferable.
Detailed description of the invention
Fig. 1 is the knee-joint anatomy photo of blank group A in the embodiment of the present invention 1.
Fig. 2 is the knee-joint anatomy photo of model group B in the embodiment of the present invention 1.
Fig. 3 is the knee-joint anatomy of administration group C (araboxylan sulfonate sodium takes orally group) in the embodiment of the present invention 1
Photo.
Fig. 4 is the knee joint of administration group D (araboxylan sulfonate sodium subcutaneous injection group) in the embodiment of the present invention 1
Dissect photo.
Fig. 5 is administration group E in the embodiment of the present invention 1 (celecoxib and Glucosamine Sulphate take orally group) knee joint solution
Cut open photo.
Fig. 6 is that each group test rabbit knee cartilage glucuronic acid content compares in the embodiment of the present invention 1.
The HE that Fig. 7 is blank group A in the embodiment of the present invention 2 dyes one of cartilage slices.
Fig. 8 is that the HE of blank group A in the embodiment of the present invention 2 dyes the two of cartilage slices.
Fig. 9 is that the HE of blank group A in the embodiment of the present invention 2 dyes the three of cartilage slices.
Figure 10 is that the HE of blank group A in the embodiment of the present invention 2 dyes the four of cartilage slices.
The HE that Figure 11 is model group B in the embodiment of the present invention 2 dyes one of cartilage slices.
Figure 12 is that the HE of model group B in the embodiment of the present invention 2 dyes the two of cartilage slices.
Figure 13 is that the HE of model group B in the embodiment of the present invention 2 dyes the three of cartilage slices.
Figure 14 is that the HE of model group B in the embodiment of the present invention 2 dyes the four of cartilage slices.
The HE that Figure 15 is administration group C (bagasse source) in the embodiment of the present invention 2 dyes one of cartilage slices.
Figure 16 is that the HE of administration group C (bagasse source) in the embodiment of the present invention 2 dyes the two of cartilage slices.
Figure 17 is that the HE of administration group C (bagasse source) in the embodiment of the present invention 2 dyes the three of cartilage slices.
Figure 18 is that the HE of administration group C (bagasse source) in the embodiment of the present invention 2 dyes the four of cartilage slices.
The HE that Figure 19 is administration group D (corn peel source) in the embodiment of the present invention 2 dyes one of cartilage slices.
Figure 20 is that the HE of administration group D (corn peel source) in the embodiment of the present invention 2 dyes the two of cartilage slices.
Figure 21 is that the HE of administration group D (corn peel source) in the embodiment of the present invention 2 dyes the three of cartilage slices.
Figure 22 is that the HE of administration group D (corn peel source) in the embodiment of the present invention 2 dyes the four of cartilage slices.
The HE that Figure 23 is administration group E (birch source) in the embodiment of the present invention 2 dyes one of cartilage slices.
Figure 24 is that the HE of administration group E (birch source) in the embodiment of the present invention 2 dyes the two of cartilage slices.
Figure 25 is that the HE of administration group E (birch source) in the embodiment of the present invention 2 dyes the three of cartilage slices.
Figure 26 is that the HE of administration group E (birch source) in the embodiment of the present invention 2 dyes the four of cartilage slices.
Figure 27 is that each group test rabbit knee cartilage glucuronic acid content compares in the embodiment of the present invention 2.
Specific embodiment
The principle and features of the present invention will be described below with reference to the accompanying drawings, and the given examples are served only to explain the present invention, and
It is non-to be used to limit the scope of the invention.
Embodiment 1: different modes of administration to treatment rabbit knee inflammation influence and with the comparison of marketed drugs
Osteoarthritis is a kind of chronic, progressive, degenerative joint disease, clinically with arthralgia, deformation and activity
With the characteristics of limited, pathological manifestations are under articular cartilage softening, fibrosis, ulcer, cartilage reduction, the hardening of subchondral bone, cartilage
Bone cyst etc..Administration mode has an important influence the performance of drug effect, this experiment investigates Arabic wood by different modes of administration
Glycan sulfonate is oral and the influence to treatment of arthritis is subcutaneously injected, and by commercial oral pharmaceutical composition (celecoxib and sulphur
Sour Glucosamine) with araboxylan sodium sulfonate it is compared research.
1. material
1.1. experimental animal
Adult New Zealand White Rabbit 40,2.8 ± 0.2kg of weight, half male and half female, sub-cage rearing.
1.2. main agents
PBS powder (China fir in Beijing), dehydrated alcohol (Chinese medicines group), (Hebei day has 0.9% sodium chloride injection at medicine company
Limit company), paraformaldehyde (the raw work in Shanghai), haematoxylin dye liquor (Nanjing is built up), eosin stain (the green skies), resinene (state
Medicine group), bamboo source araboxylan (the fine member in Guilin), Celebret (Pfizer), Glucosamine Sulphate glue
Capsule (Epson object), chloraldurate, carbazole, sulfur trioxide pyridine etc. (Shanghai Aladdin).
1.3. key instrument
Disposable syringe (Chengdu Xinjin Shi Feng Medical Devices Co., Ltd.), electronic balance (Precisa12A, Switzerland),
Centrifuge tube (Shanghai Wo Hong Biotechnology Co., Ltd), depyrogenation ultrapure water system (Millipo3150, the U.S.), high pressure sterilization
Pot (Shanghai Sai Yang), instrument for microsurgery packet (Shanghai Surgical Operation Equipment Factory), pipettor (Eppendorf, Germany), hot electric pressing
Power steam disinfecting apparatus (Shaoxing Medical Apparatus and Instruments General Factory), thermostatic drying chamber (Shanghai leap medical apparatus and instruments factory), inverted microscope (Japan
Olympus), slicer (Leca) is opened up piece machine (Leca), is baked piece machine (Leca), UV-vis spectroscopy light
Degree meter (upper Nereid section).
1.4. araboxylan sulfonate sodium is prepared
Preparation method: raw material bamboo powder araboxylan (its monosaccharide composition are as follows: the molar ratio of xylose and arabinose is weighed
It for 17:1) 45g, is placed in reaction kettle together with 210g sulfur trioxide pyridine complex, 750mL DMA, the condition of stirring is added
Under be warming up to 55 DEG C, cooling down after reaction 1 hour.41g NaOH is dissolved in water, reaction system is added, it will be anti-after being sufficiently stirred
Mixture is answered to be added in the ethyl alcohol of 2L, sufficiently rear filtering to be precipitated.Filter residue is dissolved in water, NaOH adjusts pH neutrality, ethyl alcohol is added to analyse
Out, it filters, repeats water-soluble alcohol precipitation operation twice.Products therefrom is dissolved in deionized water, in carrying out ultrafiltration in membrane separator, is retained
Molecular weight is 600Da, and gained trapped fluid is freeze-dried, obtains the araboxylan sodium sulfonate in bamboo source, specific structure
Formula is as follows:
2. experimental section
2.1. experimental group
Adult New Zealand White Rabbit 40,2.8 ± 0.2kg of weight, half male and half female is equally divided into 5 groups, sub-cage rearing.Point
Group situation is as follows:
A: normal blank group
B: model group
C: araboxylan sulfonate sodium takes orally group
D: araboxylan sulfonate sodium subcutaneous injection group
E: celecoxib and Glucosamine Sulphate take orally group
2.1. adaptive feeding
Day alternates with night by animal house 12h-12h, and protection animal only drinks water, feeds, 23-25 DEG C of temperature, enters after a week real
It tests.
2.2. arthritis model is constructed
B, C, D, E group construct arthritis model.
Rabbit weighing to be tested, is put in fixed on experimental bench after anesthesia is satisfied, drawing right rear leg is in right knee joint to stretch position,
Hospital gauze wraps up 3-4 layers (alleviation gypsum, which directly contacts skin, leads to necrosis, ulcer), the package braking of gypsum cloth, gypsum outer layer
Bandage is twined, keeps rabbit to move freely after the completion of package, braking time 6 weeks.
2.3. grouping administration
Administration group C is by (being dissolved in drinking-water) the dosage administration in 10mg/kg/ days of araboxylan sulfonate sodium；Administration group D is
Modeling leg local subcutaneous injects the administration of araboxylan sulfonate sodium 6mg/kg dosage (normal saline at aqueous solution, often
Zhou Erci)；Administration group E was by celecoxib 7mg/kg/ days (mixing in feed after stripping capsule) and Glucosamine Sulphate 15mg/
(being dissolved in drinking-water after stripping capsule) dosage administration in kg/ days；A, B group is not administered；Successive administration surrounding.
2.4. materials
Animal is put to death, and knee joint is taken, and femoral lateral condyle articular surface cartilage is taken to give over to glucuronic acid content test on a small quantity, remaining is entire
Joint carries out decalcification after 4% (v/v) paraformaldehyde is fixed, and histotomy is used for after the completion of decalcification.
2.5. paraffin sample is made, the method is as follows:
(1) it after drawing materials, is cleaned with PBS, 4% paraformaldehyde is fixed.
(2) it is dehydrated: tissue is immersed in 75% (v/v) ethyl alcohol, 85% (v/v) ethyl alcohol, 95% (v/v) ethyl alcohol two with this
It is secondary in secondary, 100% (v/v) ethyl alcohol, each 10 minutes.
(3) transparent: dewatered tissue is successively immersed in the mixing of 50% (v/v) ethyl alcohol and 50% (v/v) dimethylbenzene
It is dimethylbenzene 10 minutes, secondary 10 minutes in solvent.
(4) saturating wax: the tissue that transparent processing is crossed is soaked in the paraffin of thawing 3 hours.
(5) it embeds.
(6) it is sliced: with slicer by the histotomy in paraffin mass at the thin slice of 5 μ m-thicks, and being laid on anticreep slide.
(7) it bakes piece: slice being placed on 55 DEG C of roasting piece machines, is tightly attached to tissue on anticreep slide.
2.6. paraffin section de-waxing rehydration:
(1) it dewaxes: paraffin section being soaked in respectively dimethylbenzene 10 minutes, secondary, 50% (v/v) ethyl alcohol and 50% (v/
V) mixed solvent of dimethylbenzene 10 minutes.
(2) rehydration: by the paraffin section after dewaxing be soaked in 100% (v/v) ethyl alcohol is secondary, 95% (v/v) ethyl alcohol is secondary,
In 85% (v/v) ethyl alcohol, 75% (v/v) ethyl alcohol, each 5 minutes.
(3) distilled water cleans 3 times, every time 2 minutes.
2.7. hematoxylin eosin staining:
(1) haematoxylin dye liquor dyes 10 minutes.
(2) tap water rinses extra dyeing liquor, and about 5 minutes.
(3) distilled water washs one time again.
(4) microscopy, if haematoxylin hyperchromatosis can carry out a point decoloration with 1% acidic alcohol, it is extra in cytoplasm to slough
Haematoxylin dyeing liquid.
(5) eosin stain dyes 30 seconds.
(6) the extra dye liquor of tap water flushing, about 3 minutes.
(7) distilled water washs one time again.
2.8. it is dehydrated mounting:
(1) 95% (v/v) ethanol dehydration 2 minutes is secondary.
(2) dimethylbenzene transparent 5 minutes it is secondary.
(3) resinene mounting.
(4) microscopy, nucleus au bleu, cytoplasm is at red or pink.
2.9. glucuronic acid content measures in cartilage
(1) the articular cartilage sample left and taken, accurately weighs 5mg, is placed in test tube, is diluted to 10% hydrochloric acid 1mL dropwise
It is added, sealing is heated 2 hours in 90 DEG C of water-baths, is cooled to room temperature, and the 0.1% carbazole test solution colour developing of 0.2mL is added, stands 2
Hour after in ultraviolet-uisible spectrophotometer test 530nm absorbance.
(2) drafting of standard curve, takes the standard uronic acid sample 1mL of various concentration respectively, and method as above measures extinction
Degree, and draw fitting concentration-absorbance curve, as standard curve.
(3) content of uronic acid in sample is calculated using the fit equation of standard curve.
3. experimental result
3.1. animal behavior is observed
Activity condition is as follows before animal is put to death:
A group: test rabbit rear leg is strong, freedom of movement.
B group: test rabbit does not like activity, and modeling leg is not involved in body-support, or even only supports by foreleg with sitting posture movement.
C group: test rabbit does not like activity, does not have significant difference with model group.
D group: the accidental modeling leg support of discrete trial rabbit shows slightly powerless, remaining strong activity of test rabbit modeling leg is normal.
E group: test rabbit activity is not limited by modeling leg, but the support of visible modeling leg still shows slightly powerless, hence it is evident that is not so good as blank
Group.
3.2.Pelletier scoring
After test rabbit joint tissue is removed, knee joint sample gross examination of skeletal muscle is carried out, is commented by Pelletier point system
Point, see Table 1 for details, and as a result see Table 2 for details.Part knee joint sample photo is shown in Fig. 1-5.
1 Pelletier point system of table
2 Pelletier appraisal result of table
Note: a and A group have significant difference (P < 0.01), and b and B group have significant difference (P < 0.01), and c and C group have significantly
Sex differernce (P < 0.01), d and D group have significant difference (P < 0.01).
As can be seen from Table 2, have the difference of conspicuousness between administration group D and administration group E and other each groups, and araboxylan
Between sodium sulfonate oral administration group C and model group B and there was no significant difference.And araboxylan sulfonate sodium is subcutaneously injected
D group effect is substantially better than the E group of oral marketed drugs combination.
3.3. histopathology is evaluated
Blank group A group articular cartilage surface is smooth, and no crack or defect, four-layer structure is clear, and damp line is complete.Model group B
Group layer of articular cartilage is thinning, and four-layer structure is difficult to differentiate, and surface layer cartilage defect is serious, and cartilage cell is disorganized, and cell is a large amount of
The poly- hyperplasia of cluster and fibrosis, damp line destroy serious.Administration group C group layer of articular cartilage is thinning, and four-layer structure is difficult to differentiate, and surface layer is soft
Bone defect is serious, and cartilage cell is disorganized, the poly- hyperplasia of a large amount of clusters of cell and fibrosis, and damp line destroys serious.Administration group D group is closed
Cartilage cell's queueing discipline is saved, level is substantially complete, the accidental Cells Depletion of surface part, and damp line is still complete.Administration group E group joint
Cartilage layers are thinning, and four-layer structure is difficult to differentiate, and surface layer cartilage defect is serious, and cartilage cell is disorganized, and cell has the poly- hyperplasia of cluster
And fibrosis phenomenon, damp line destroy more serious.
3.4. in cartilage uronic acid content
The forfeiture of articular cartilage proteoglycan is the important symbol that osteoarthritis occurs, and proteoglycan and uronic acid contain in cartilage
Amount can more normally significantly reduce, and the severity of the changes of contents and osteoarthritis of uronic acid is in inverse ratio in cartilage.If Fig. 6 is each
The comparison of rabbit knee cartilage glucuronic acid content is tested in group, wherein oral administration group C and other each groups have significant difference
(P < 0.05), administration group D and model group B, administration group C and E have significant difference (P < 0.01), administration group E and other each groups
There is significant difference (P < 0.01).
4. conclusion
Pass through glucuronic acid content measurement in animal behavior observation, dissection and Histopathological Studies and cartilage, it is found that
The pharmacodynamic results of each administration group are that D group is better than E group, and E group is better than C group again.Araboxylan sodium sulfonate subcutaneous administrations group
It is all substantially better than other each groups in each index, is not only better than the oral administration group of araboxylan sodium sulfonate, but also
It is substantially better than marketed drugs combination (celecoxib and Glucosamine Sulphate).From cartilage glucuronic acid content substantially go up and
Histopathological Studies can speculate that araboxylan sodium sulfonate has significant facilitation to the nutrition and regeneration of damaged cartilage；
Dissection gross examination of skeletal muscle can be found that the test rabbit joint sample inflammation of araboxylan sodium sulfonate drug administration by injection disappears substantially
It loses；Araboxylan sodium sulfonate known to both sides experimental evidence is closed by diminishing inflammation and promoting cartilage nutrition to play bone
Save scorching therapeutic effect.And subcutaneous injection is substantially better than the macromolecular structure that oral administration illustrates araboxylan sodium sulfonate
It is unfavorable for alimentary canal absorption or is possible to the decomposition by the various enzymes of alimentary canal and enteric microorganism, it cannot be with effective component
Form reaches affected part.
Although influence of the xylan sulfonate of the different plant origins of embodiment 2, different structure to rabbit knee inflammation is almost
All contain various types of xylans in all terrestrial plant cell membranes, but the structural difference of xylan itself is extremely aobvious
It writes or even completely different, therefore the purpose of the present embodiment is the xylan sulfonate pair of the different plant origins of research, architectural difference
The influence for treating rabbit osteoarthritis illustrates the relationship of macromolecular structure and its therapeutic effect different in raw material.
1. material
1.1. experimental animal
Adult New Zealand White Rabbit 40,2.8 ± 0.2kg of weight, half male and half female, sub-cage rearing is equally divided into 5 groups, point
It is not:
A: normal blank group
B: model group
C: one administration group of sample
D: two administration group of sample
E: three administration group of sample
1.2. main agents
In addition to raw material sources are different, remaining with embodiment 1, specifically: bagasse source, corn peel source araboxylan
(the fine member in Guilin), birch source xylan (Sigma-Aldrich).
1.3. key instrument
With embodiment 1.
1.4. araboxylan sulfonate sodium is prepared
Method with embodiment 1, raw material be respectively bagasse source araboxylan (branched structure, xylose and I
The molar ratio of uncle's sugar is 11.2:1), the maize peel source araboxylan (molar ratio of branched structure, xylose and arabinose
For 3.5:1) and birch source xylan (direct coupled structure, no arabinose), alkali used is KOH.Prepared by the above raw material
Sulfonic acid potassium salt number consecutively be sample one (bagasse source), sample two (corn peel source) and sample three (birch source), three
The structural formula difference of sample is as follows:
Sample one:
Sample two:
Sample three:
2. experimental section
In addition to all subcutaneous injections of administration mode, including injection dosage other test methods etc. with embodiment 1
It is identical.
3. experimental result
3.1. animal behavior is observed
Activity condition is as follows before animal is put to death:
A group: test rabbit rear leg is strong, and activity is freely.
B group: test rabbit does not like activity, and modeling leg is not involved in body-support, or even only supports by foreleg with sitting posture movement.
C group: test rabbit modeling leg is strong, and activity is normal, does not have significant difference with blank group.
D group: test rabbit modeling leg is strong, and activity is normal, does not have significant difference with blank group.
E group: the accidental modeling leg support of discrete trial rabbit shows slightly powerless, remaining strong activity of test rabbit modeling leg is normal.
3.2.Pelletier scoring
After test rabbit joint tissue is removed, knee joint sample gross examination of skeletal muscle is carried out, is commented by Pelletier point system
Divide (with embodiment 1), as a result see Table 3 for details.
3 Pelletier appraisal result of table
Note: a and A group have significant difference (P < 0.01), and b and B group have significant difference (P < 0.01), and c and D group have significantly
Sex differernce (P < 0.05).
Seen from table 3, relative to model group, the damage of the sample articular cartilage of three administration groups is all smaller, illustrates three
Sample can have the function that promotion articular cartilage is restored, and especially D group behaves oneself best.There was only D, E bis- between three administration groups
There are significant difference (P < 0.05) for group.
3.3. histopathology is evaluated
Histotomy result is shown in Fig. 7-Figure 26.Wherein, blank group A group articular cartilage surface is smooth, no crack or defect, and four
Layer is clear in structure, and damp line is complete, is detailed in Fig. 7-Figure 10.Model group B group layer of articular cartilage is thinning, and four-layer structure is difficult to differentiate, table
Layer cartilage defect is serious, and cartilage cell is disorganized, the poly- hyperplasia of a large amount of clusters of cell and fibrosis, and damp line destroys seriously, is detailed in figure
11- Figure 14.Administration group C group articular chondrocytes queueing discipline, level is substantially complete, and surface part cell is complete, and damp line is still complete
It is whole, it is detailed in Figure 15-Figure 18.Administration group D group articular chondrocytes queueing discipline, level is substantially complete, and surface part cell is complete,
Damp line is still complete, is detailed in Figure 19-Figure 22.Administration group E group articular chondrocytes queueing discipline, level is substantially complete, surface part
Accidental Cells Depletion, the visible agger line of damp line still intact part sample, is detailed in Figure 23-Figure 26.
3.4. in cartilage uronic acid content
The comparison that rabbit knee cartilage glucuronic acid content is tested in each group, is detailed in Figure 27.Wherein model group and other each groups
Have significant difference (P < 0.01), administration group C and blank group A and administration group E have significant difference (P < 0.05), administration group D
There is significant difference (P < 0.01) with administration group E.
4. conclusion
From animal activity condition as it can be seen that three administration groups show apparent arthritis treatment effect；Dissection is substantially
Observation and pathological section can find that D group experimental animal articular cartilage recovery is best, and C group is taken second place, and E group is relatively slightly poor；
The result of glucuronic acid content and dissection and case slice result are almost the same in cartilage, are equally that the articular cartilage of administration group D is extensive
Multiple situation is best, and E group restores slightly slow.It can be seen that the source of raw material xylan and structure influence obviously osteoarthritis curative effect,
Corn peel and bagasse are monocotyledon in the source of three xylans, and birch is dicotyledon.Wherein, branched chain type
And the araboxylan effect in the most abundant corn peel source of branch is best, straight chain type and the birch source wood without arabinose
Glycan effect is most weak, so being conducive to be promoted using the araboxylan potassium sulfonate of the high xylan preparation of arabinose content
Drug effect.
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention, it is all in spirit of the invention and
Within principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Claims (10)
1. application of the araboxylan sulfonate in the drug of preparation treatment osteoarthritis.
2. application according to claim 1, which is characterized in that the general structure of the araboxylan sulfonate are as follows:
In formula, the summation of x, y and z are 1-19；The 2- that side chain sulfonation arabinose passes through 1- oxygen glycosidic bonds and main chain sulfonation xylose
Or/and 3- connections, and the molar ratio of the two is 1:2-1:20；Side chain miscellaneous sugar passes through 1- oxygen glycosidic bonds and main chain sulfonation xylose
2- or/and 3- connection；Miscellaneous sugar in the sulfonation miscellaneous sugar is glucose, glucuronic acid, 4- methyl glucose uronic acid, gala
One of sugar, rhamnose, galacturonic acid, xylose are a variety of.
3. application according to claim 1, which is characterized in that the preparation method of the araboxylan sulfonate, packet
Include following steps:
Step 1: by araboxylan and sulfonating agent 1:(1-5 in molar ratio), being placed in reaction kettle together, obtain the first mixing
Object；
Step 2: the ratio of 100mL polar organic solvent being added according to araboxylan described in every (1-8) g step 1, to step
Polar organic solvent is added in 1 the first obtained mixture, to 55 DEG C -95 DEG C, reaction is down to after 1-3 hours for warming while stirring
Room temperature obtains the second mixture；
Step 3: lye is added into the second mixture that step 2 obtains, sulfonating agent described in the alkali in the lye and step 1
Molar ratio is 1:1, and after being sufficiently stirred, obtained reaction mixture is added to 2-5 times of volume of polar organic solvent described in step 2
Ethyl alcohol in, it is to be precipitated sufficiently after filtering, obtain filter residue；
Step 4: according to every 10g filter residue be dissolved in (50-100) mL water ratio carry out it is water-soluble, with lye tune identical in step 3
Saving pH value is 7, and the ethyl alcohol that 2-5 times of volume of above-mentioned water is added carries out alcohol precipitation, and sediment is filtered, and repeats above-mentioned water-soluble, alcohol precipitation, mistake
The operation of filter twice, is collected last sediment, is dissolved in deionized water, crosses ultrafiltration membrane, gained trapped fluid is freeze-dried, i.e.,
Obtain the araboxylan sulfonate.
4. application according to claim 3, which is characterized in that the sulfonating agent is sulfur trioxide trimethylamine compound, three
One in sulfur oxide triethylamine compound, sulfur trioxide pyridine complex or sulfur trioxide N,N-dimethylformamide compound
Kind；The polar organic solvent is one of dimethyl sulfoxide, N,N-dimethylformamide or DMAC N,N' dimethyl acetamide；Institute
Stating room temperature is 20-25 DEG C；The lye is NaOH solution, KOH solution, Ca (OH)2One of solution；Point of the ultrafiltration membrane
Sub- interception is 600Da.
5. application according to claim 3, which is characterized in that the araboxylan is extracted from monocotyledon.
6. application according to claim 5, which is characterized in that the monocotyledon be sugarcane, bamboo, corn, oat,
One of reed, rice, wheat are a variety of.
7. application according to claim 1, which is characterized in that the drug is with a effective amount of araboxylan sulfonic acid
Salt is active constituent, in addition the pharmaceutical preparation that pharmaceutically acceptable carrier and/or auxiliary material are prepared.
8. application according to claim 1, which is characterized in that the dosage form of the drug is injection.
9. application according to claim 8, which is characterized in that the injection includes aqueous solution or water composite solution.
10. application according to claim 8, which is characterized in that the administration mode of the drug is subcutaneous injection, intramuscular note
It one of penetrates, be injected intravenously or is a variety of.
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| WO2020147350A1 (en) * | 2019-01-15 | 2020-07-23 | 广西壮族自治区中国科学院广西植物研究所 | Use of araboxylan sulfonate in preparation of medicine for treating osteoarthritis |
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| CN109793751B (en) * | 2019-01-15 | 2021-02-26 | 广西壮族自治区中国科学院广西植物研究所 | Application of arabinoxylan sulfonate in preparation of medicine for treating osteoarthritis |
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