CN110403947A - Application of the Purmorphamine in pharmacy - Google Patents
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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
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
- A61K31/5375—1,4-Oxazines, e.g. morpholine
- A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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Abstract
The application that the invention discloses Purmorphamine in pharmacy, including inhibiting the application in osteoclast differentiation and maturation drug in preparation.Present invention finds Purmorphamine to be able to suppress osteoclast differentiation, can be applied to titanium metal particles and mediates in the prevention and treatment of bone dissolution, and can play more obvious action.
Description
Technical field
The present invention relates to the purposes of Purmorphamine, more particularly to the purposes in pharmaceutical field.
Background technique
China human mortality aging is on the rise, and China 60 years old or more population is more than 2.1 hundred million at present, it is contemplated that increases to the year two thousand thirty
3.55 hundred million are added to, the year two thousand forty increases to 4.10 hundred million, and associated aged's health problem situation is very severe.Wherein bone with
Joint disease seriously affects the life span and quality of patient, some researches show that occur Hip Fracture after in 1 year, about 50% patient
Disable, 20% patient will die of various complication, and the quality of life of sufferer is decreased obviously.Artificial joint replacement (total
Joint arthroplasty, TJA) it will the joint prosthesis made of metal, high molecular polythene or ceramics surgical technic
In Using prosthesis body, instead of the support and motor function of diseased joints, reach pain caused by the causes of disease such as relief from osteoarthritis, fracture
The motor function in joint is restored in pain and part, is that hip joint, knee joint fracture compound comminuted and terminal option share bone are involved in current treatment
Necrosis, rheumatoid arthritis, the preferred treatment method of osteoarthritis.It is significantly increased and prosthese service life in TJA amount for surgical
Under extended background, prosthese aseptic loosening is as hip joint, the postoperative long term most important complication of knee prosthesis, it has also become
The principal element of Using prosthesis service life is influenced, while being also that TJA is postoperative and needing the main reason for overhauling.
After being implanted into prosthese by surgical technic, the fine motion of prosthese-bone interface and joint motions abrasion can lead to prosthese-bone
Between generate the wear particle being largely made of polymethyl methacrylate (PMMA), polyethylene and titanium particle.These are along vacation
The wear particle being distributed between body-bone not only increases the frictional force between prosthese and bone, is more also easy to produce new wear particle, shape
Wear particle can be swallowed at vicious circle, and by the associated immune cells of representative of macrophage, is secreted out of largely such as
The inflammatory factors such as TNF-α, IL-1, IL-6, IL-8 promote the differentiation and bone resorption function of osteoclast (Ostroclast, OC),
So as to cause Periprosthetic bone loss even prosthetic loosening.In addition, prosthesis wear particle can obviously inhibit mescenchymal stem cell and
Osteoblast secretes 1 Collagen Type VI, resorption lacunae and reduces bone mineralising, and can be obviously promoted it and secrete IL-8, GM-CSF, M-
The proinflammatory cell factor for promoting osteoclastic differentiation such as CSF, RANKL.
Clinical practice in recent years confirms, by improving bone cement technology, improving prosthetic material and design, emphasis operator doctor
The modes such as operation skill, drug application, aseptic loosening can delay or prevent.Wherein, anti-osteoporosis treats, is oral
Calcium tablet and bisphosphonates are able to suppress osteoclast function；Wear-resisting prosthetic material is selected, such as high crosslinked polyethylene gasket,
Ceramic prosthese etc., can be relieved wear particle precipitation and adjoint osteoclast cell activation.
Although researcher and clinician are implanted by design Custom made stem, using wear-resistant material, improvement
Mode etc. is to reduce the generation of prosthese aseptic loosening, but less effective, and wound is big, effect is poor, complication is more, costly
Prosthese revision procedure be still aseptic prosthetic loosening primary treatment regimen, still lack for aseptic prosthetic loosening this together
Send out the drug of the effective therapeutic effect of disease.For the prevention and treatment of prosthese aseptic loosening, clinically mainly by particular point in time
Take that the secondary osteoclast of bisphosphonates prevention wear particle largely activates prevention, revision procedure thoroughly removes bone water
Mud and limitans treat the complication, there is the point that is difficult to take the time, diphosphonate use causes complication, operation wound big etc.
Disadvantage.
Purmorphamine binds directly and activates the compound of Smoothened (Smo) memebrane protein as one kind, can swash
Hedgehog signal path living.In the past document report, Purmorphamine can be obviously promoted osteoblast differentiation, but its
Important participation component osteoclast another on bone metabolism process influences the corresponding research of not yet expansion.
Summary of the invention
The purpose of the present invention is to provide the new applications of Purmorphamine, i.e., the new opplication in pharmacy.
The present invention provides Purmorphamine to inhibit the application in osteoclast differentiation and maturation drug in preparation.
The present invention also provides Purmorphamine to inhibit correlated characteristic gene in osteoclast atomization in preparation
Application in Nfatc1, C-fos, Ctsk, Acp5, Oscar, Dc-stamp, Atp6v0a3, Atp6v0d2 expression drug.
The present invention also provides Purmorphamine to inhibit JNK signal path in osteoclast atomization living in preparation
Application in chemical drug object.
The present invention also provides Purmorphamine to prepare titanium metal particles caused by prevention and treatment osteoclast activity increases extremely
Mediate answering in the dissolution drug of bone caused by bone dissolution, osteoporosis, rheumatoid arthritis, caput femoris necrosis, bone metastaes
With.
Beneficial effects of the present invention:
(1) in osteoclast Analytical Chemical Experiment, the amount of osteoclast in the case where Purmorphamine concentration is 0.5 μM of processing
And area significantly reduces, 1 μM handles lower inhibition level and becomes apparent from, and 2 μM of whens are differentiated to form without osteoclast substantially.
(2) Quantitative reverse transcription PCR as the result is shown osteoclast correlated characteristic gene expression with
The raising of Purmorphamine concentration and in concentration dependent reduce.
(3) protein immunoblotting as the result is shown mouse monokaryon macrophage after 2 μM of Purmorphamine are pre-processed,
JNK access is obviously suppressed in the signal path for participating in osteoclast differentiation under RANKL stimulation, and JNK phosphoric acid turns to p-JNK
Process obviously weakens, and AKT, NF- κ B, p38, ERK signal path activation process do not weaken.At 1,3 day, long time-histories was osteoclastic simultaneously lures
It leads in protein immunoblotting experiment, the important transcription factor of osteoclast differentiation is participated in 2 μM of Purmorphamine groups
The relatively control of Nfatc1, C-fos protein expression amount is obviously inhibited.
(4) model, Micro-CT detection, 3D reconstruction, bone parameter point are dissolved using titanium metal particles building mouse skull bone
Purmorphamine can significantly alleviate bone course of dissolution as the result is shown for analysis etc., and diaphysis fraction BV/TV, bone density BMD are more positive
Control group obviously increases, and skull bone dissolves lacuna number, the decline of area conspicuousness.
Therefore, Purmorphamine can be applied in the prevention and treatment that titanium metal particles mediate bone to dissolve, and can play more aobvious
The effect of work.
Detailed description of the invention
Fig. 1 (a)~(c) is respectively cell of the Purmorphamine to mononuclear macrophage after cultivating 48h, 72h and 96h
Toxicity figure；
Fig. 2 is influence diagram of the Purmorphamine to the osteoclastic atomization of mononuclear macrophage, wherein (a)~(d) is respectively
For tartaric-resistant figure；(e), (f) be respectively osteoclast number and area-graph；
Fig. 3 is the influence that Purmorphamine generates the osteoclastic atomization characteristic F-actin ring of mononuclear macrophage
Figure, wherein (a) be phalloidine fluorescent staining detection figure, (b), (c) be respectively osteoclast number and area-graph；
Fig. 4 is the influence diagram of Purmorphamine differentiation osteoclastic to mononuclear macrophage and bone resorption ability, wherein (a)
It is (b) area-graph of Bone resoiption pit for the SEM image of ox bone piece Bone resoiption pit；
Fig. 5 is that Quantitative reverse transcription PCR detects Gli1, Nfatc1, C-fos, Ctsk, Acp5, Oscar, Dc-
The expression figure of the characterizing genes such as stamp, Atp6v0a3, Atp6v0d2；
Fig. 6 is the signal of interest access that protein immunoblotting detects that Purmorphamine is related to osteoclast differentiation
Middle AKT, NF- κ B, p38, p65, JNK, ERK phosphorylation activation are horizontal；
Fig. 7 is that protein immunoblotting detects Purmorphamine to NFATc1, c-fos turns in osteoclast differentiation
Record the protein expression level of the factor；
Fig. 8 (a) scans the image of mouse skull (b) for Micro-CT, (c) is the bone Parameter analysis result of mouse skull.
Specific embodiment
The present invention will be further explained below with reference to the attached drawings and specific examples, so that those skilled in the art can be with
It more fully understands the present invention and can be practiced, but illustrated embodiment is not as a limitation of the invention.
Embodiment 1:Purmorphamine inhibits osteoclast differentiation and without obvious cytotoxicity
(1) isolation and culture of Primary bone marrow source mononuclear macrophage BMMs
C57BL/6 mouse one of 5 to 8 weeks is taken, 75% medicinal alcohol impregnates 3 minutes after drawing neck to put to death, in ultra-clean work
Mouse double lower limb is dissected in platform, separation skin, fascia and knee joint hip joint joint capsule obtain mouse femur and shin bone, be placed in
It is added in the Micro-Organism Culture Dish of sterilizing phosphate buffered saline solution in advance.Proximal femur and shin bone lower end are cut off using sterile scissors,
Marrow is isolated using 10000rpm brief centrifugation method, 2ml erythrocyte cracked liquid is added, marrow is resuspended, it is red to stand cracking in 5 minutes
Cell.Then horizontal centrifugal (1000rpm, 5 minutes) at room temperature, discard after liquid plus 10ml M-CSF containing 30ng/ml it is complete
Alpha MEM culture solution, soft piping and druming are mixed, are moved in 10cm steril cell culture dish, in 37 DEG C, 5%CO2It is incubated in incubator
It educates.Liquid is changed to remove not adherent heteroproteose cell within 2nd day, continues culture until the mononuclear macrophage of derived from bone marrow is long to 80% close
Degree or more.
(2) digestion and counting of Primary bone marrow source mononuclear macrophage
Culture medium is discarded, sterile phosphate buffer saline solution is added 3ml pancreatin after cleaning twice and digests 10 minutes, under microscope
The culture medium 1ml containing serum is added after visible cell slightly shrinkage, slowly collects culture solution after soft piping and druming cell, room temperature is lauched
Flat centrifugation (1000rpm, 5 minutes).It discards supernatant, the culture medium containing serum in right amount is added, cell is resuspended, gently piping and druming mixes；Together
When take 100 μ l cell suspensions to manage to EP, 1900 μ l, 0.4% trypan blue dye liquor is added, microscopically observation in 3min counts meter
The cell number of color is not caught in the number big lattice of plate 4 by dye liquor.Cell suspension cell number/ml=(4 big lattice total number of cells/4) ×
Extension rate (20 times) × 104.(note: dead cell can be contaminated for blue, and living cells will not be colored)
(3) cytotoxicity (Fig. 1) of the CCK-8 method detection Purmorphamine to mononuclear macrophage
Mononuclear macrophage kind is entered into 96 orifice plates, cell density is 5 × 103Cells/well, totally 33 holes (1 control of every plate
Group, 9 various concentration dosing groups, every group of 3 multiple holes), Purmorphamine maximum concentration is 32 μM；Culture medium is containing 30ng/
The complete alpha MEM of the M-CSF of ml cultivates 48,72,96 hours respectively, changes liquid every other day.Until after corresponding time point, every hole
10 μ l CCK-8 reagents are added, takes out under dark condition in cell incubator incubation, is fullyd shake simultaneously in microplate reader after forty minutes
Absorbance value (competent cell number is more, and absorbance is higher) at 450nm is measured, experimental result shows that Purmorphamine is dense
Degree is proliferated without obvious inhibiting effect mononuclear macrophage at 2 μM or less.
(4) tartaric-resistant detects Purmorphamine to the osteoclastic atomization of mononuclear macrophage
Influence (Fig. 2)
If control group and 3 drug-treated groups, every group of 3 multiple holes, by mononuclear macrophage kind in 24 orifice plates (10 ×
104Cells/well), culture medium is changed to RANKL containing 50ng/ml every other day, the complete Alpha MEM of 30ng/ml M-CSF is cultivated
Liquid, and 0.5,1,2 μM of Purmorphamine is separately added into drug-treated group, control group is added and 2 μM
The dimethyl sulfoxide solvent of Purmorphamine group equivalent changes liquid every other day, cultivates 6 days, when control group has obvious mature hypertrophy
It is terminated when osteoclast formation, after culture medium is sopped up, phosphate buffered saline solution is cleaned 1 time, and 4% paraformaldehyde 500ul/ is added
Fixed cell 20 minutes of hole carries out tartaric-resistant using kit, takes pictures under inverted microscope the simultaneously later period
The quantity and spreading area of the osteoclast more than 3 nucleus, the tartaric-resistant positive are calculated, it is clear
Purmorphamine has inhibiting effect to osteoclast differentiation.It is 0.5 μM that experimental result, which is shown in Purmorphamine concentration,
It handles lower amount of osteoclast and area significantly reduces, 1 μM handles lower inhibition level and becomes apparent from, substantially without osteoclast at 2 μM
It is differentiated to form.
(5) phalloidine fluorescent staining detects Purmorphamine to the osteoclastic atomization characteristic of mononuclear macrophage
The influence (Fig. 3) that F-actin ring generates
If control group and 3 drug-treated groups, every group of 3 multiple holes, by mononuclear macrophage kind in 24 orifice plates (10 ×
104Cells/well), culture medium is changed to RANKL containing 50ng/ml every other day, the complete Alpha MEM of 30ng/ml M-CSF is cultivated
Liquid, and 0.5,1,2 μM of Purmorphamine is separately added into drug-treated group, control group is added and 2 μM
The dimethyl sulfoxide solvent of Purmorphamine group equivalent changes liquid every other day, cultivates 6 days, when control group has obvious mature hypertrophy
It is terminated when osteoclast formation, after culture medium is sopped up, phosphate buffered saline solution is cleaned 1 time, and 4% paraformaldehyde 500ul/ is added
Fixed cell 20 minutes of hole, phosphate buffered saline solution exhausts after cleaning 2 times.300 hole μ l/ 0.1%Triton X-100 is added to incubate
It educates 5 minutes, phosphate buffered saline solution exhausts after cleaning 2 times, and 1% bovine serum albumin(BSA), 300 hole μ l/ is incubated for and exhausts after five minutes, adds
Enter 1% phalloidine to be incubated for 20 minutes, DAPI is redyed after exhaustion, finally removes dyeing liquor and phosphate buffered saline solution is added,
It sets and takes pictures under fluorescence microscope.Experimental result prompt Purmorphamine can inhibit to concentration dependent mononuclear macrophage broken
The generation of characteristic F-actin ring in bone atomization.
(6) ox bone piece Bone resoiption pit evaluation Purmorphamine differentiation osteoclastic to mononuclear macrophage and bone resorption energy
The influence (Fig. 4) of power
Fresh bulls backbone dense bone is taken, electricity consumption is sawed into about 2.5cm × 2.5cm × 2.5cm size bone block, then
Bone block is put into hard tissue slicing machine-cut into about 80 μm of thickness of bone thin slice, the punch of 96 orifice plate aperture sizes beats thin osteocomma
It is advisable at can just be put in 96 orifice plates, the osteocomma cut is impregnated 30 minutes in 70% alcohol.Take 12 osteocommas, In
Aseptic condition is displaced downwardly to drying in 50ml centrifuge tube, is placed in 96 orifice plates after Alpha MEM cleaning 3 times of 5ml serum-free are added
In dry, ultraviolet irradiation sterilizing.Every hole kind enters mononuclear macrophage (5 × 103Cells/well), culture medium is changed to contains every other day
The complete Alpha MEM culture solution of 50ng/ml RANKL, 30ng/ml M-CSF, and be separately added into drug-treated group 0.5,
1, the dimethyl sulfoxide solvent with 2 μM of Purmorphamine group equivalent is added in 2 μM of Purmorphamine, control group, every other day
Liquid is changed, is cultivated 6 days.Osteocomma front and back sides are taken out and mark, in osteocomma front, toothbrush gently brushes surface osteoclast and fragment,
After drying osteocomma, metal spraying, scanning electron microscope are taken pictures.Experimental result show ox bone piece Bone resoiption pit quantity and area with
The raising of Purmorphamine concentration and in concentration dependent reduce.
Embodiment 2:Purmorphamine activates the important transcription factor Gli1 of Hedgehog signal path and inhibits osteoclastic thin
The expression of correlated characteristic gene in born of the same parents' atomization
(1) cell processing and RNA collect separation
If control group and 3 drug-treated groups, every group of 3 multiple holes, by mononuclear macrophage kind (40 × 10 in 6 orifice plates4
Cells/well), culture medium is changed to the complete Alpha MEM culture solution of RANKL containing 50ng/ml, 30ng/ml M-CSF every other day,
And 0.5,1,2 μM of Purmorphamine is separately added into drug-treated group, control group is added and 2 μM of Purmorphamine
The dimethyl sulfoxide solvent of group equivalent, changes liquid every other day, cultivates 6 days, when control group has obvious mature loose osteoclast formation
When terminate.Total serum IgE is extracted using RNA extraction agent box, after measuring concentration, takes in 500ng RNA reaction system and synthesizes cDNA.It should
Reaction system includes reverse transcription buffer, primer, dNTP, reverse transcriptase, DEPC water, RNA template (10 μ l of total volume), synthesis knot
Sample using ultrapure water is diluted to 100 μ l after beam, and to be stored in -80 DEG C of refrigerators spare.
(2) Quantitative reverse transcription PCR detects the expression (Fig. 5) of correlated characteristic gene in osteoclastic atomization
Quantitative reverse transcription PCR selects 20 μ l reaction systems, and each system is separately added into 2 μ l of cDNA, upstream and downstream
Each 0.25 μ l of primer, 10 μ l of SYBR Green qPCR Mix, ddH2O 7.5μl.On Bio-Rad CFX96 instrument according to
Lower reaction condition carries out: 95 DEG C of initial denaturation 3 minutes, then recycling 40 times within 30 seconds within 10 seconds, 60 DEG C for 95 DEG C, is judged according to solubility curve
Whether each system reaction is normal, and the software analysis result finally provided using fluorescence quantitative PCR instrument obtains CT value, uses 2- Δ
The analysis of Δ CT method and more each sample directly correspond to the mrna expression amount of gene and for statistical analysis.Experimental result is shown
The expression of the important transcription factor Gli1 of Hedgehog signal path is in concentration dependant with the raising of Purmorphamine concentration
Property rise, osteoclast correlated characteristic gene Nfatc1, C-fos, Ctsk, Acp5, Oscar, Dc-stamp, Atp6v0a3,
The expression of Atp6v0d2 is reduced with the raising of Purmorphamine concentration in concentration dependent.
Embodiment 3:Purmorphamine inhibit osteoclast atomization in JNK signal path activation and important turn
Record the expression of factor NFATc1, c-fos
(1) albumen sample process and extraction
Engineered protein is handled in short-term: by mononuclear macrophage kind (40 × 10 in 6 orifice plates4Cells/well, 30ng/ml M-CSF
Complete Alpha MEM culture solution), be divided into control group drug-treated group, every other day before extracting albumen 4 hours respectively at drug
2 μM of Purmorphamine are added in reason group, and control group is added equivalent dimethyl sulfoxide and is pre-processed.Two groups are separately added into
50ng/ml RANKL was to activate osteoclast to break up associated signal paths, respectively at 0,5,15,30,60 minute collection albumen sample
This.
Engineered protein is handled when long: by mononuclear macrophage kind (40 × 10 in 6 orifice plates4Cells/well, 30ng/mlM-CSF
Complete Alpha MEM culture solution), be divided into control group drug-treated group, overnight after culture medium is changed to containing 50ng/ml
The complete Alpha MEM culture solution of RANKL, 30ng/ml M-CSF, and 2 μM are added into drug-treated group
Purmorphamine, control group are added the dimethyl sulfoxide solvent of equivalent, liquid are changed every other day, respectively at 1,3 day collection albumen sample
This.
Albumen sample extraction and concentration mensuration: it wait cultivate to corresponding time point, quickly inhales and abandons supernatant, 4 DEG C of phosphoric acid buffers
Washed with saline solution 1 time, 6 orifice plates are placed on ice, and every hole is added prepares the RIPA containing protease inhibitors, inhibitors of phosphatases in advance
100 μ l of cell pyrolysis liquid is sufficiently cracked 20 minutes after piping and druming on ice, and lysate moves to 1.5ml EP pipe, in advance 4 DEG C of pre-cooling centrifugations
Machine is centrifuged 13000rpm, 15 minutes, collects supernatant into new EP pipe, carry out every pipe sample egg using BCA protein concentration kit
White concentration mensuration, -80 DEG C of refrigerators save.
(2) protein immunoblotting detection Purmorphamine to the osteoclast signal of interest access that is related to of differentiation and
The influence (Fig. 6, Fig. 7) of transcription factor activator and expression
In the signal of interest access being related to using protein immunoblotting detection engineered protein sample osteoclast differentiation in short-term
AKT, NF- κ B, p38, JNK, ERK, p65 phosphorylation activation are horizontal, specify Purmorphamine by influencing a certain signal path
Inhibit osteoclast differentiation.NFATc1, c-fos turn in the osteoclastic atomization of engineered protein sample when further long by detection simultaneously
The variation of factor protein expression is recorded, further to verify the effect of Purmorphamine regulation osteoclast differentiation.Experiment
Purmorphamine can regulate and control NFATc1, c-fos transcription factor expression by influencing JNK protein phosphorylation process as the result is shown
To inhibit osteoclast to break up.
Embodiment 4:Purmorphamine prevents and treats titanium metal particles and mediates bone dissolution
(1) drug dissolution, mice group and modeling post-processing: 2%DMSO+30%PEG 300+5%Tween 80+ is prepared
ddH2The solution that Purmorphamine is made into final concentration of 125 μ g/ml is added in O solvent.C57BL/6 mouse 24, random point
It is 4 groups, every group 6, blank control group conventinal breeding is only given above-mentioned solvent intraperitoneal injection after positive controls modeling success,
Low concentration medicine group and high concentration medicine group after modeling success, give respectively daily Purmorphamine 0.25mg/kg and
1mg/kg intraperitoneal injection is primary.
(2) titanium metal particles mouse skull bone dissolves model: blank control group implements sham-operation, and skull surface is cut in anesthesia
Skin and after removing skull surface connective tissue, skin suture does not smear titanium metal particles.Positive controls, low concentration drug
Group and high concentration medicine group implement modeling operation, after anaesthetizing successfully, aseptically cut cranium skin, remove skull table
Face connective tissue takes 30mg titanium particle to be uniformly applied to braincap bone surface, skin suture.It is post-processed through above-mentioned modeling, after 14 days
De- neck is put to death, and solution takes cranium and is put in after 4% paraformaldehyde fixes 2 days, is put into 75% alcohol and is saved.
(3) Micro-CT scans each group skull and by operations such as 2D building, 3D reconstructions, obtains skull three-dimensional image
And analyze related bone amount parameter (Fig. 8).There is obvious bone dissolution phenotype, model compared with negative control group in positive controls as the result is shown
It is successfully constructed, while skull bone dissolution phenotype is obviously changed compared with positive controls in low concentration medicine group and high concentration medicine group
Kind, Purmorphamine can significantly alleviate the process that titanium metal particles mediate bone dissolution.
Embodiment described above is only to absolutely prove preferred embodiment that is of the invention and being lifted, protection model of the invention
It encloses without being limited thereto.Those skilled in the art's made equivalent substitute or transformation on the basis of the present invention, in the present invention
Protection scope within.Protection scope of the present invention is subject to claims.
Claims (4)
1.Purmorphamine inhibits the application in osteoclast differentiation and maturation drug in preparation.
2.Purmorphamine preparation inhibit osteoclast atomization in correlated characteristic gene Nfatc1, C-fos, Ctsk,
Application in Acp5, Oscar, Dc-stamp, Atp6v0a3, Atp6v0d2 expression drug.
3.Purmorphamine inhibits the application in osteoclast atomization in JNK signal path pharmacological activation in preparation.
Titanium metal particles caused by 4.Purmorphamine preparation prevention and treatment osteoclast activity increases extremely mediate bone dissolution, sclerotin
Bone caused by loose, rheumatoid arthritis, caput femoris necrosis, bone metastaes dissolves the application in drug.
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| KR20100000387A (en) * | 2008-06-24 | 2010-01-06 | 한경대학교 산학협력단 | Complex containing purmorphamine derivative for promoting formation of bone |
| EP2455079A2 (en) * | 2009-07-17 | 2012-05-23 | Korea Research Institute of Bioscience and Biotechnology | Composition for the prevention or treatment of bone diseases comprising colforsin daropate |
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2019
- 2019-09-12 CN CN201910867050.2A patent/CN110403947A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20100000387A (en) * | 2008-06-24 | 2010-01-06 | 한경대학교 산학협력단 | Complex containing purmorphamine derivative for promoting formation of bone |
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