CN105943531B - Bis- (adjacent aminobenzene sulfydryl) applications of the butadiene in pharmacy of 1,4- diamino -2,3- dicyanos -1,4- - Google Patents
Bis- (adjacent aminobenzene sulfydryl) applications of the butadiene in pharmacy of 1,4- diamino -2,3- dicyanos -1,4- Download PDFInfo
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- CN105943531B CN105943531B CN201610293848.7A CN201610293848A CN105943531B CN 105943531 B CN105943531 B CN 105943531B CN 201610293848 A CN201610293848 A CN 201610293848A CN 105943531 B CN105943531 B CN 105943531B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/275—Nitriles; Isonitriles
- A61K31/277—Nitriles; Isonitriles having a ring, e.g. verapamil
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
- A61K31/375—Ascorbic acid, i.e. vitamin C; Salts thereof
Abstract
The invention discloses a kind of drug of promotion tendon injury healing, the drug contains bis- (the adjacent amino-benzene oxygen sulfydryl) butadiene of medicinal 2,3 dicyano Isosorbide-5-Nitrae of a effective amount of Isosorbide-5-Nitrae diamino.Another kind promotes the drug of tendon injury healing to be made of 1,4 diamino, 2,3 dicyano 1,4 bis- (adjacent amino-benzene oxygen sulfydryl) butadiene, vitamin C and pharmaceutically acceptable non-active ingredients.The healing of kneecap tendon can be effectively facilitated by being also proved drug of the present invention using internal rat kneecap tendon injury model.The drug has many advantages, such as at low cost, convenient drug administration, good effect, safe without toxic side effect compared with other therapeutic schemes, is the drug of more satisfactory promotion tendon injury.
Description
Technical field
The invention belongs to tendon injury medicine technical fields, more particularly to a kind of Isosorbide-5-Nitrae-diamino -2,3- dicyan
Bis- (adjacent aminobenzene sulfydryl) applications of the butadiene in pharmacy of base -1,4-.
Background technology
The damage of tendon and ligament is common orthopaedics problem.The incidence of tendon injury just increases year by year, traces it to its cause
Mainly movement is participated in more and more a middle-aged persons or strenuous exercise is related.By taking heel string as an example:Rupture of Achilles tendon incidence is (6-
37)/105, wherein 75% is related with sports, this does not include other damages of heel string also.Tendon injury can be divided into acute
And chronic injury, reason include inherent or external factor, either internal factor or internal and external factor combination can all cause flesh
Tendon injury.Tendon injury occurs mainly in movement and workplace, this normally results in affected part pain, stiff and tendon intensity damage
Wound.Heel string and kneecap tendon injury are most common two kinds of movements tendon injuries.In the relevant factor of tendon injury, age and gender
Factor is most common factor, and the incidence of male is compared with women higher.Damage caused by work accounts for Causes of Acute Traumatic tendon
The 24.9% of damage.Since indolence after tendon injury, and impaired tendon can be made to be restored to without a kind of operation plan
Normal institutional framework and mechanical strength, so tendon injury is still clinically one serious and pendent ask
Topic.
Tendon injury is difficult to repair.The generation of healing process of tendons can be divided into three phases.In the initial inflammatory reaction stage,
Red blood cell and inflammatory cell, especially bite neutrophil cell, enter damage location.Then, Tenocyte cell gradually will move to wound
Mouthful, the synthesis of type III collagen is also activated in this stage.Several days are spent, proliferative phase starts.Type III collagen is in this stage
Synthesis reach peak value, and by continued for several weeks.Remodelling phase starts after about six weeks, and during which cell composition reduction, collagen subtract
It is few, and start to synthesize glycosaminoglycan.However, the biochemistry of tendon tissue and mechanical performance are still in those intact tendons after healing
It differs greatly.
Tendon injury will produce a large amount of unsound state, and no matter which type of care method be used, caused by it
Deformity may periods of months.Many physics modes are used in the Case management of disorder of tendon.Although however, these modes by with
In routine clinical, only a small number of controlled clinical trials have been completed.Most of evidences still fall within preclinical research, and
Sometimes the result is that tool is controversial.In the Achilles tendinitis model by collagen enzyme induction of rat, extracorporeal impact wave therapy is demonstrate,proved
It is bright to pass through induction TGF β 1 (Transforming growth factor-beta, transforming growth factor-β) and IGF-I
The expression of (Insulin-like growth factor 1, type-1 insulin like growth factor) promotes the healing of tendon.Also,
Rat Achilles Tendon inflammation model, 17 hertz of pulsed magnetic fields and electromagnetic field can improve the frequency of arrangement of collagen fibers.In addition, in recent years with
Organizational engineering and biomedical emerging development, certain cell factors and growth factor such as TGF β and IGF-I, Yi Jiji
Because the MSC (mesenchymal stem cell, mescenchymal stem cell) for the treatment of and used in tissue engineering has been used for tendon injury
Treatment, and show tempting foreground.But the problem of these growth factors is expensive and easy degradation, to patient with
Carry out prodigious financial burden.
Invention content
The object of the present invention is to provide a kind of bis- (the adjacent aminobenzene sulfydryl) fourths two of 1,4- diamino -2,3- dicyanos -1,4-
Application of the alkene in pharmacy.
The technical solution that the present invention solves above-mentioned technical problem is as follows：A kind of 1,4- diamino -2,3- dicyanos -1,4- is double
(the adjacent aminobenzene sulfydryl) application of butadiene in pharmacy, the drug contain medicinal a effective amount of Isosorbide-5-Nitrae-diamino -2,3- bis-
Bis- (the adjacent aminobenzene sulfydryl) butadiene of cyano -1,4-.Preferably, the drug contains 5 μM~15 μM of Isosorbide-5-Nitrae-diamino -2,3-
Bis- (the adjacent aminobenzene sulfydryl) butadiene of dicyano -1,4-.It is highly preferred that the drug contains 10 μM of Isosorbide-5-Nitrae-diamino -2,3-
Bis- (the adjacent aminobenzene sulfydryl) butadiene of dicyano -1,4-.
Bis- (adjacent aminobenzene sulfydryl) applications of the butadiene in pharmacy of another Isosorbide-5-Nitrae-diamino -2,3- dicyano-Isosorbide-5-Nitrae -,
By bis- (the adjacent aminobenzene sulfydryl) butadiene of 1,4- diamino -2,3- dicyanos -1,4-, vitamin C and pharmaceutically acceptable
Non-active ingredient forms.Preferably, the drug contains 5 μM~15 μM of the bis- (neighbours of Isosorbide-5-Nitrae-diamino -2,3- dicyano-Isosorbide-5-Nitrae -
Aminobenzene sulfydryl) butadiene.It is highly preferred that Isosorbide-5-Nitrae-diamino -2,3- dicyano-Isosorbide-5-Nitrae-that the drug contains 10 μM is bis- (adjacent
Aminobenzene sulfydryl) butadiene.
Exist in bis- (the adjacent aminobenzene sulfydryl) butadiene of 1,4- diamino -2,3- dicyanos -1,4- as described above of the invention
In application in pharmacy, pharmaceutically acceptable non-active ingredient is：Pharmaceutically acceptable diluent pharmaceutically may be used
The excipient of receiving and pharmaceutically acceptable supporting agent.
Exist in bis- (the adjacent aminobenzene sulfydryl) butadiene of 1,4- diamino -2,3- dicyanos -1,4- as described above of the invention
In application in pharmacy, the drug is any one clinically-acceptable peroral dosage form, injection type or exterior-applied formulation.
Exist in bis- (the adjacent aminobenzene sulfydryl) butadiene of 1,4- diamino -2,3- dicyanos -1,4- as described above of the invention
In application in pharmacy, the drug is tablet, capsule, granule, pill, pill, liquid preparation, soft extract, hangs
Floating agent, dispersant, syrup, suppository, gelling agent, aerosol, patch.
The present invention proposes to promote the theory and thinking of tendon injury Cure Study On with U0126.Its main function principle is
U0126 can promote the mescenchymal stem cell in rat and the tendon source of people to break up to Tenocyte cell, to repair damaged tissues.
U0126 single preparationss of ephedrine proposed by the present invention, the wherein optimal dose of U0126 are 10 μM, and can be according to damaged area
Size is done suitable for adjustment, fool proof.This drug can be prepared into injection for clinical application by the modern crafts of pharmacy.Face
Injection is primary every three days when bed application.For containing ascorbic composition, 25 μM of VC optimal doses, the best agent of U0126
Amount is 10 μM.
Bis- (adjacent aminobenzene sulfydryl) applications of the butadiene in pharmacy of 1,4- diamino -2,3- dicyanos -1,4- include with
Lower step：
Step 1, it weighs, Isosorbide-5-Nitrae-bis- (adjacent aminobenzenes of diamino -2,3- dicyano-Isosorbide-5-Nitrae-is weighed according to predetermined formulation proportioning
Sulfydryl) butadiene, vitamin C and pharmaceutically acceptable non-active ingredient；
Step 2, the component weighed in step 1, is uniformly mixed that form medicine material spare by mixing；
Step 3, the medicine material obtained in step 2 is prepared into clinically-acceptable dosage form.
The present invention is also proved this pharmaceutical preparation using internal 1/3 damage model of rat kneecap tendon can effectively facilitate kneecap tendon
Healing.The drug has at low cost, convenient drug administration, good effect, safe without toxic side effect etc. excellent compared with other therapeutic schemes
Point is the drug of more satisfactory promotion tendon injury.
Description of the drawings
Fig. 1 is U0126 to mescenchymal stem cell Toxic test results；
Fig. 2-1 to 2-6 is that Rat Mesenchymal Stem Cells break up situation to Tenocyte cell for 24 hours for U0126 processing；
Fig. 3-1 to 3-6 is that U0126 handles 72h Rat Mesenchymal Stem Cells to Tenocyte cell differentiation situation；
Fig. 4 is that U0126 handles 72h human mesenchymal stem cells to Tenocyte cell differentiation situation；
Fig. 5 is rat kneecap tendon one third damage model schematic diagram；
Fig. 6 is rat kneecap tendon healing result；
Fig. 7-1 to 7-2 is rat kneecap tendon Mechanical test results.
Specific implementation mode
U0126(C18H16N6S2·1/2C2H5OH it is) a kind of MEK1/2 inhibitor of high selectivity, U0126 is acted on
The cell of TPA (12-O-Tetradecanoylphorbol 13-acetate, terephthalic acid (TPA)) stimulations, by blocking MAPK
(Mitogen-activated protein kinases, mitogen-activated protein kinase) signal transmits, and significantly inhibits c-Fos
Up-regulation with c-Jun mRNA and protein level inhibit to reach 50-80%, lead to AP-1 (Activator protein 1, activation
Sub- albumen -1) transcriptional activity is suppressed, and IC50 is 0.96 μM.10 μM of U0126 by inhibiting ERK2 rather than ERK1 phosphorylations,
Significantly inhibit cell death.U0126 is suffered from the mouse of allergic asthma by 30mg/kg dosage intraperitoneal injections, is had significant
Antiphlogistic effects, there are dose dependents for this effect.
Vitamin C (also known as L-AA) is the essential nutrients of High Primates animal and other a small number of biologies.It is anti-
Bad hematic acid can be manufactured in most organism by metabolism, but the mankind are most significant exceptions.It is most well-known
Be a lack of vitamin C and can cause scurvy.In vivo, vitamin C is a kind of antioxidant, and protection body is from freedom
The threat of base, vitamin C are also a kind of coenzyme simultaneously.Vitamin C is that antibody and collagen are formed, tissue repairing (including certain oxygen
Change reduction), the synthesis of fat, protein maintains immune function, and keeps the complete of blood vessel, and nonheme iron is promoted to inhale
The institute such as receipts is required.The pharmaceutical composition of vitamin C and U0126 compositions can promote the mesenchyma in rat and the tendon source of people dry
Cell breaks up to Tenocyte cell, to repair damaged tissues.
Bis- (the adjacent aminobenzene sulfydryl) butadiene of 1,4- diamino -2,3- dicyanos -1,4- of the first specific implementation mode
Application in pharmacy, the drug contain medicinal a effective amount of Isosorbide-5-Nitrae-bis- (adjacent aminobenzenes of diamino -2,3- dicyano-Isosorbide-5-Nitrae -
Sulfydryl) butadiene.Preferably, the drug contains 5 μM~15 μM of Isosorbide-5-Nitrae-bis- (adjacent amino of diamino -2,3- dicyano-Isosorbide-5-Nitrae -
Benzene sulfydryl) butadiene.It is highly preferred that the drug contains 10 μM of Isosorbide-5-Nitrae-bis- (adjacent amino of diamino -2,3- dicyano-Isosorbide-5-Nitrae -
Benzene sulfydryl) butadiene.
Bis- (the adjacent aminobenzene sulfydryl) butadiene of 1,4- diamino -2,3- dicyanos -1,4- of second of specific implementation mode
Application in pharmacy, by Isosorbide-5-Nitrae-diamino -2,3- dicyano-Isosorbide-5-Nitrae-bis- (adjacent aminobenzene sulfydryl) butadiene, vitamin C and doctors
Pharmaceutically acceptable non-active ingredient composition.Preferably, the drug contains 5 μM~15 μM of Isosorbide-5-Nitrae-diamino -2,3- bis-
Bis- (the adjacent aminobenzene sulfydryl) butadiene of cyano -1,4-.It is highly preferred that the drug contains 10 μM of Isosorbide-5-Nitrae-diamino -2,3- bis-
Bis- (the adjacent aminobenzene sulfydryl) butadiene of cyano -1,4-.
Above-mentioned pharmaceutically acceptable non-active ingredient is：Pharmaceutically acceptable diluent can pharmaceutically connect
The excipient and pharmaceutically acceptable supporting agent received.
Exist in bis- (the adjacent aminobenzene sulfydryl) butadiene of 1,4- diamino -2,3- dicyanos -1,4- as described above of the invention
In application in pharmacy, the drug is tablet, capsule, granule, pill, pill, liquid preparation, soft extract, hangs
Floating agent, dispersant, syrup, suppository, gelling agent, aerosol, patch.
1 liquid preparation of embodiment
Bis- (the adjacent aminobenzene sulfydryl) butadiene of 10 μM of 1,4- diamino -2,3- dicyanos -1,4- are dissolved into 10 μ l and gone
In ionized water, liquid preparation is formed.
2 liquid preparation of embodiment
By bis- (the adjacent aminobenzene sulfydryl) butadiene of 5 μM of 1,4- diamino -2,3- dicyanos -1,4- be dissolved into 10 μ l go from
In sub- water, liquid preparation is formed.
3 liquid preparation of embodiment
Bis- (the adjacent aminobenzene sulfydryl) butadiene of 15 μM of 1,4- diamino -2,3- dicyanos -1,4- are dissolved into 10 μ l and gone
In ionized water, liquid preparation is formed.
4 tablet of embodiment
Bis- (the adjacent aminobenzene sulfydryl) butadiene of Isosorbide-5-Nitrae-diamino -2,3- dicyano-Isosorbide-5-Nitrae-are taken, lactose is added, stearic acid forms sediment
Powder crosses 80 mesh sieve, and mixing, with 70% ethyl alcohol softwood, 20 mesh sieve is pelletized, dry, 16 mesh sieves, tabletting, and piece weighs about
100mg, every contains 15 μM of active constituent.
5 aerosol of embodiment
Foaming agent：Dodecyl sodium sulfate 0.9g, Ninol 0.6g, water 28.5g are uniformly mixed；
Propellant：Isopentane 4g, monochlorodifluoromethane 3g, normal butane 3g, mixing；
Bis- (the adjacent aminobenzene sulfydryl) butadiene of 1,4- diamino -2,3- dicyanos -1,4- and foaming agent and propellant are mixed
It closes, forms aerosol, the wherein a concentration of 10 μM/ml of active ingredient.
6 liquid preparation of embodiment
By 25 μM of vitamin Cs and bis- (the adjacent aminobenzene sulfydryl) butadiene of 10 μM of 1,4- diamino -2,3- dicyanos -1,4-
It is dissolved into 10 μ l deionized waters, forms liquid preparation.
7 tablet of embodiment
Bis- (the adjacent aminobenzene sulfydryl) butadiene of vitamin C, Isosorbide-5-Nitrae-diamino -2,3- dicyano-Isosorbide-5-Nitrae-are taken, lactose is added,
Stearic acid, starch cross 80 mesh sieve, mixing, with 70% ethyl alcohol softwood, the sieve granulation of 20 mesh, dry, 16 mesh sieves, tabletting, piece
100mg is weighed about, every containing 15 μM of U0126,25 μM of vitamin C.
8 aerosol of embodiment
Foaming agent：Dodecyl sodium sulfate 0.9g, Ninol 0.6g, water 28.5g are uniformly mixed；
Propellant：Isopentane 4g, monochlorodifluoromethane 3g, normal butane 3g, mixing；
Bis- (the adjacent aminobenzene sulfydryl) butadiene of 1,4- diamino -2,3- dicyanos -1,4- and foaming agent and propellant are mixed
It closes, forms aerosol, a concentration of 10 μM/ml of wherein U0126,25 μM/ml of vitamin C.
Application effect embodiment
1 material and method
The culture of the mescenchymal stem cell in 1.1 tendon sources
8 week old SD rats are taken, male, weight 200-250g.After intraperitoneal anesthesia, rats with bilateral kneecap tendon is taken out, rejects tendon
Tendon is uniformly shredded into 1mm x 1mm sizes by sheath and tendon circumferential portion, and per 100mg, the enzyme of Type I collagen containing 3mg/ml is added in tissue
(Sigma companies) digests 1h in 37 DEG C of incubators.After stopping digestion, together with direct kind of complete tendon residual tissue may do not digested
In Tissue Culture Flask, penicillin containing 100U/ml, 100U/ml streptomysins and 10% fetal calf serum is added, and (Invitrogen is public
Department) DMEM (Invitrogen companies) culture medium, be placed in 37 DEG C, 5%CO2Incubator in be incubated.Continue culture (to change for 8-10 days
The liquid period is 3 days/time), it is observed that tendon stem cell colonies are formed.After cell amplification, take P3~P8 alternative in related real
It tests.The tendon stem cell in people source is donated by Prince of Wales Hospital of Hong Kong Chinese University, is to agree under the premise of institute in contributor
The sample of acquisition, the cell are similarly incubated in DMEM culture mediums, and 10% fetal calf serum and 100U/ml moulds has been added
Element, 100U/ml streptomysins, are placed in 37 DEG C, 5%CO2Incubator in be incubated.
1.2 drug
PBS, U0126 are purchased from Sigma Chemical companies of the U.S..
Research of the 1.3 U0126 preparations to mescenchymal stem cell toxicity
To verify toxicity of the U0126 preparations to mescenchymal stem cell of various concentration, we measure the work of cell with mtt assay
Power.Mescenchymal stem cell is inoculated in 96 orifice plates, after being used in combination the U0126 preparations of various concentration to handle 24 and 72 hours, is being trained
The MTT that 5mg/ml is added in base is supported, is read after being incubated 4h.
1.4 U0126 preparations promote mescenchymal stem cell to break up at tendon
To prove the influences broken up at tendon to mesenchymal stem cells of U0126, we by Rat Mesenchymal Stem Cells in vitro
For 24 hours and 72h with U0126 inductions, or by people source tendon stem cell pharmaceutical composition processing 72h, while we are with TGF β
As positive controls.Total serum IgE is extracted from cell with the small extraction reagent kits of RNeasy later, uses MMLV inverse after measuring RNA concentration
Transcriptase carries out reverse transcription according to the operating guidance of offer.Intracellular tendon mark of correlation base is tested and analyzed with real-time quantitative PCR
The expression situation of cause, such as Tenomodulin, Scleraxis, Collagen type I and II, Tenascin C,
Decorin, Biglycan and Fibromodulin etc..
The foundation of 1.5 rat kneecap tendon injury models and U0126 treatments
4 monthly age SPF grade male SD rats 20 (Hong Kong Chinese University Experimental Animal Center provide) are provided, weight (254 ±
20)g.Adaptable fed 10d.The rat kneecap tendon defect model having been had built up according to my laboratory exposes the kneecap tendon of rat,
One third (about 1mm) among longitudinal sectional excision.Rat is randomly divided into PBS (phosphate buffered saline solution) control groups and drug therapy
Group (10 μM of U0126 of 10 μ l), every group 10.Every three days PBS or medicine, Gong Jisan are injected in kneecap tendon defect location
It is secondary.All animals freely take the photograph water and ingest (standard feed, calcic 1.33%, phosphorus 0.95%), in 25 28 DEG C of room temperature, humidity
It is fed in 7580% Clean Facility.It draws materials after 3 weeks, sample segment carries out mechanical test (n=6), and remainder kneecap tendon carries out
Fixing process carries out paraffin section and histological observation such as HE dyeing and immunohistochemical staining etc. later.
1.6 mechanical test
The method of mechanical test carries out in accordance with the following methods.First, complete kneecap tendinous tissue is isolated, later simultaneously with two
Knife blade together cuts away original tendon tissue on periphery and retains newborn tendon tissue.Later, by the both ends of kneecap tendon
It is fixed on mechanics test system (Hounsfield H25KS mechanics testers, Tinius Olsen companies, Britain).Mechanical test
Parameter used is 40 mm/mins, and the pulling force of pre-loaded 0.1N, when the maximum that reaches newborn tendon tissue and can bear
Tendon is broken when power, and machine can record the curve of power and displacement at this time.The newborn muscle tendon groups measured according to curve and in advance
The cross-sectional area knitted, we calculate maximum, force, and (unit is N/mm2).And Young's modulus is then calculated according to slope of a curve
It arrives.
1.7 statistics
Measurement data mean ± standard deviationIt indicates, simultaneously using SPSS16.0 software row ANOVA variance analyses
It is examined with LSD-t and carries out comparison among groups, P<0.05 it is believed that have statistical significance.
2 results
2.1 U0126 are to mescenchymal stem cell nontoxicity
First, we measure toxicity of the U0126 to Rat Mesenchymal Stem Cells of various concentration by mtt assay.As a result
Show within the scope of 0.1-20 μM of concentration, U0126 does not have significant toxic effect to cell, shows that the drug is peace to cell
Complete (Fig. 1).
Fig. 1 shows the U0126 of various concentration to mescenchymal stem cell nontoxicity.Mescenchymal stem cell is through various concentration
After U0126 is handled 24 and 72 hours, the vigor (n=5) of cell is measured with mtt assay.
2.2 U0126 can start mescenchymal stem cell to be broken up at tendon
According to the cytotoxicity experiment of U0126, we finally select 10 μM of optimal dosages as tendon injury treatment.For
The influence that detection U0126 breaks up mescenchymal stem cell at tendon, mescenchymal stem cell is handled for 24 hours respectively through 10 μM of U0126
After 72h, the total serum IgE for extracting cell carries out reverse transcription and real-time quantitative PCR (real time PCR) detection.The tendon of detection
It includes colI (Collagen type I, I-type collagen), scx (Scleraxis), mkx to break up relevant marker gene
(Mohawk Homeobox), tnmd (Tenomodulin), fmod (Fibromodulin) and decorin (Fig. 2-3).As a result table
Bright, the expression of colI, Scx, mkx, tnmd and fmod of U0126 processing groups have significant change.
Meanwhile above-mentioned experiment is repeated in our employment source mescenchymal stem cells, i.e., after handling 72h with 10 μM of U0126, into
Row real-time quantitative PCR detection tendon breaks up the expression of relevant marker gene.The result shows that 10 μM of U0126 compare blank control
Group and TGF-β group, can preferably improve the expression (Fig. 4) of colI, mkx, tnmd and decorin, with Rat Mesenchymal Stem Cells
Experimental result coincide substantially.Break up to tendon these results indicate that U0126 is a potential promotion mescenchymal stem cell
Effective micromolecular compound.
Fig. 2-1 to Fig. 2-6 U0126 start Rat Mesenchymal Stem Cells to be broken up to Tenocyte cell.Mescenchymal stem cell is through 10
After μM U0126 is handled 24 hours, the total serum IgE for extracting cell carries out quantitative PCR detection (n=3).By further testing pair
Ascorbic effect is added to be detected, U0126 combinations VC starts Rat Mesenchymal Stem Cells to be broken up to Tenocyte cell.Between fill
For matter stem cell after 10 μM of U0126 and 25 μM of vitamin Cs are handled 24 hours, the total serum IgE for extracting cell carries out quantitative PCR detection
(n=3).The result shows that 10 μM of U0126 are relatively used alone in the varied in ploidy that Rat Mesenchymal Stem Cells break up to Tenocyte cell
Processing is higher by 32%~40%.It is above-mentioned statistics indicate that, vitamin C can further increase the using effect of U0126, promote cell
Differentiation.
Fig. 3-1 to Fig. 3-6 U0126 start Rat Mesenchymal Stem Cells to be broken up to Tenocyte cell.Mescenchymal stem cell is through 10
After μM U0126 is handled 72 hours, the total serum IgE for extracting cell carries out quantitative PCR detection (n=3).Mescenchymal stem cell is through 10 μM
After U0126 and 25 μM of vitamin C is handled 72 hours, the total serum IgE for extracting cell carries out quantitative PCR detection (n=3).The result shows that
The varied in ploidy that Rat Mesenchymal Stem Cells break up to Tenocyte cell be relatively used alone 10 μM of U0126 processing be higher by 30%~
41%.It is above-mentioned statistics indicate that, vitamin C can further increase the using effect of U0126, promote cell differentiation.
Fig. 4 U0126 start human mesenchymal stem cell to be broken up to Tenocyte cell.Human mesenchymal stem cell is through 10 μM of U0126
After processing 72 hours, the total serum IgE for extracting cell carries out quantitative PCR detection (n=3).Human mesenchymal stem cell is through 10 μM of U0126
After being handled 72 hours with 25 μM of vitamin Cs, the total serum IgE for extracting cell carries out quantitative PCR detection (n=3).The result shows that the human world
The varied in ploidy that mesenchymal stem cells are broken up to Tenocyte cell is relatively used alone 10 μM of U0126 processing and is higher by 28%~36%.It is above-mentioned
Statistics indicate that vitamin C can further increase the using effect of U0126, promote cell differentiation.
2.3 U0126 promote healing process of tendons
Can facilitation be played to the healing for damaging tendon further to verify U0126, we establish rat kneecap tendon
One third damage model (Fig. 5).Period injects rat 10 μM of U0126 every three days, and dosage is 10 μ l, co-injection
Three times, control group injects PBS buffer solution.Materials carry out mechanical test and histological inspection after three weeks.HE is dyed and Scx immune groups
Weave chemistry dyeing proves that the kneecap tendon healing state of U0126 treatments is significantly better than control group (Fig. 6).Also, the result of mechanical test
Show that U0126 can significantly improve the maximum, force that treatment group's rat kneecap tendon can bear, and Young's modulus is not notable with control group
Difference (Fig. 7-1 to Fig. 7-2).In further testing, 10 μM of U0126 and 25 μM of dimensions are injected every three days to rat
Raw element C, co-injection three times, materials progress mechanical test after three weeks, mechanical test the result shows that U0126 combination vitamin C energy
Further increase the maximum, force that treatment group's rat kneecap tendon can bear.
Each technical characteristic of above-mentioned disclosure is not limited to disclosed and other feature combination, and those skilled in the art are also
Other combinations between each technical characteristic can be carried out according to the purpose of invention, be subject to realize the present invention purpose.
Claims (10)
1.1,4- diamino -2,3- dicyanos -1,4- is double（Adjacent aminobenzene sulfydryl）Butadiene is preparing promotion tendon injury healing
Application in drug.
2. application according to claim 1, which is characterized in that the drug contains 5 μM~15 μM of Isosorbide-5-Nitrae-diamino-
2,3- dicyanos -1,4- is double（Adjacent aminobenzene sulfydryl）Butadiene.
3. application according to claim 2, which is characterized in that the drug contains 10 μM of Isosorbide-5-Nitrae-diamino -2,3- bis-
Cyano -1,4- is double（Adjacent aminobenzene sulfydryl）Butadiene.
4. application according to claim 1, which is characterized in that bis- by Isosorbide-5-Nitrae-diamino -2,3- dicyano-Isosorbide-5-Nitrae -（Adjacent ammonia
Base benzene sulfydryl）Butadiene, vitamin C and pharmaceutically acceptable non-active ingredient composition.
5. application according to claim 4, which is characterized in that the drug contains 5 μM~15 μM of Isosorbide-5-Nitrae-diamino-
2,3- dicyanos -1,4- is double（Adjacent aminobenzene sulfydryl）Butadiene.
6. application according to claim 5, which is characterized in that the drug contains 10 μM of Isosorbide-5-Nitrae-diamino -2,3- bis-
Cyano -1,4- is double（Adjacent aminobenzene sulfydryl）Butadiene.
7. according to claim 4-6 any one of them applications, which is characterized in that pharmaceutically acceptable non-active ingredient
For：Pharmaceutically acceptable diluent, pharmaceutically acceptable excipient and pharmaceutically acceptable supporting agent.
8. application according to claim 1, which is characterized in that the drug is any one clinically-acceptable oral agents
Type, injection type or exterior-applied formulation.
9. application according to claim 1, which is characterized in that the drug is tablet, capsule, granule, pill, decocts
Paste, suspending agent, syrup, suppository, gelling agent, aerosol, patch.
10. application according to claim 1, includes the following steps：
Step 1, it weighs, it is bis- to weigh Isosorbide-5-Nitrae-diamino -2,3- dicyano-Isosorbide-5-Nitrae-according to predetermined formulation proportioning（Adjacent aminobenzene sulfydryl）
Butadiene, vitamin C and pharmaceutically acceptable non-active ingredient；
Step 2, the component weighed in step 1, is uniformly mixed that form medicine material spare by mixing；
Step 3, the medicine material obtained in step 2 is prepared into clinically-acceptable dosage form.
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| CN201610293848.7A CN105943531B (en) | 2016-05-05 | 2016-05-05 | Bis- (adjacent aminobenzene sulfydryl) applications of the butadiene in pharmacy of 1,4- diamino -2,3- dicyanos -1,4- |
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