CN108939085A - Bone targeting medicine-carried system and preparation method thereof - Google Patents
Bone targeting medicine-carried system and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of Bone targeting medicine-carried systems and preparation method thereof, are polysaccharide-graphene quantum dot gel composites of BMP-2 modification, and BMP-2 is fairlead or target head, and polysaccharide-graphene quantum dot gel is pharmaceutical carrier；BMP-2 as fairlead or target head is attached to polysaccharide-graphene quantum dot gel surface.Bone targeting medicine-carried system of the invention not only possesses the excellent performance of three-dimensional netted stereochemical structure, large specific surface area, medicament slow release that its persursor material polysaccharide-graphene quantum dot has, and tests prove that the gel medicine-carried system Bone targeting after BMP-2 modification, and Bone targeting selectivity is high.Therefore medicine-carried system of the invention is the superior drug carrier with Bone targeting.
Description
Technical field
The present invention relates to a kind of Bone targeting medicine-carried systems and preparation method thereof.
Background technique
Bone target medicine delivery system by with hydroxyapatite have specific affinity as fairlead or the base of target head
Group and pharmaceutical carrier two parts composition.Drug can enter adhesive bond or in a manner of being wrapped up by carrier body,
And it discharges in vivo.Bone target medicine delivery system is applied to clinic, is had for bone metabolic diseases such as treatment osteoporosis
Significance.In general, bone target medicine delivery system is compared with common form of medication, bone target medicine delivery system energy
It is enough in reaching medicine effective concentration in therapentic part in the short period, to improve bioavilability, reduce medication dosage and
Frequency reduces systemic side effects.
BMPs(Bone Morphogenetic Protein) it is one group of heterodimeric protein, the two specific chain groups combined by cysteine
At being hydrophobicity, acidoglycoprotein.BMP-2 is considered as the strongest BMPs hypotype of induced osteogenesis activity, the effect to cell
It is chiefly to facilitate cell differentiation, can obviously induce undifferentiated BMSCs to cartilage cell, osteoblast differentiation, and then is induced new
Bon e formation.Its half-life period in vivo is shorter, cannot continue to maintain effective local concentration, therefore be limited in clinical application.
At present to the application of BMP-2, mainly its Cell differentiation inducing activity utilized is to induce the characteristic of new bone formation.
Such as 103169950 A(application number 201310080970.2 of Chinese patent literature CN) disclose a kind of BMP-2
Sustained-release micro-spheres and preparation method thereof, microsphere matrices ingredient are chitosan and sodium dextran sulfate, and the drug of package is BMP-2, packet
Envelope rate reaches the holding of 75%~90%, BMP-2 protein active and the sustained release period is long.It, will in order to solve the problems, such as BMP-2 half-life short
Spansule is made in it.
In another example 101835493 A(application number 200880107383.5 of Chinese patent literature CN) disclose a kind of amphiphilic
Compound between property polymer and the osteogenic protein for belonging to BMP family, the amphiphilic polymer is by passing through hydrophobic substituent
The hy-drophilic polysaccharide skeleton being functionalized with hydrophilic radical is constituted.Likewise, the amphiphilic polymer-BMP compound is used for
Bon e formation is induced in vivo.
In the document, polysaccharide be selected from hyaluronan, alginates, chitosan, polygalacturonic acid, chondroitin sulfate,
Glucan, cellulose.
For chitosan therein, chitosan is the deacetylated product of chitin, with unique biocompatibility, life
The properties such as object degradability, antibacterial, nontoxic and non-immunogenicity are widely used in medicine and other fields.Studies have shown that chitosan is logical
Amphiphilic polymer can be formed by crossing structural modification, then by physically trapping drug, can solubilized insoluble medicine and raising drug substance stable
Property, side effect is reduced, bioavilability is improved, and carries wide medicine range, stable structure, with excellent tissue permeability, is one
Kind has potential targetable drug carriers.
Graphene quantum dot is novel zero dimension inorganic nano material, there is nontoxic, good biocompatibility, low toxicity
Property, the performances such as biodegradability and size be controllable.Because it is with very high specific surface area, the drug loading of superelevation can be possessed
Amount, therefore, graphene quantum dot is suitably applied pharmaceutical carrier field.
105477647 A(application number 201510867975.9 of Chinese patent literature CN) disclose a kind of graphene quantum
The preparation of point/chitosan xerogel and be applied to fluorescence imaging and medicament slow release.But graphene quantum dot/the chitosan is dry solidifying
Glue only has the ability for being used as carrier, and does not have Bone targeting.
Summary of the invention
Technical problem to be solved by the invention is to provide a kind of medicine-carried system and preparation method thereof with Bone targeting.
The technical solution for realizing the object of the invention is a kind of Bone targeting medicine-carried system, is polysaccharide-graphene of BMP-2 modification
Quantum dot gel composite, BMP-2 are fairlead or target head, and polysaccharide-graphene quantum dot gel is pharmaceutical carrier；As guiding
Object or the BMP-2 of target head are attached to polysaccharide-graphene quantum dot gel surface.
The polysaccharide is one of chitosan, sodium alginate, lactobionic acid, beta-cyclodextrin.
BMP-2 and polysaccharide-graphene quantum dot gel mass ratio are 1:2.5~20.
A kind of preparation method of Bone targeting medicine-carried system as described above, comprising the following steps:
1. preparing polysaccharide-graphene quantum dot gel.
2. the building of BMP-2 plasmid.
3. the preparation of BMP-2 modified polysaccharide-graphene quantum dot gel: by step 1. in prepare polysaccharide-graphene amount
Son selects gel and step, and 2. the middle BMP-2 plasmid leaching liquor centrifugation prepared after mixing, is placed in water-bath, 40 DEG C~60
After DEG C water-bath preheats 10~30min, vortex is mixed, then is placed at room temperature for, and obtains polysaccharide-graphene quantum dot-BMP-2 CDNA, from
The heart abandons supernatant, obtains chitosan-graphene quantum dot gel of BMP-2 modification after rinsing, freeze-drying.
Step 2. the building of BMP-2 plasmid when, using the genomic DNA extracted in rat peripheral blood, pass through polymerase chain
Formula reacts (PCR) and synthesizes BMP-2 gene, it is carried out genetic recombination, transformed competence colibacillus Escherichia coli, structure with pcDNA3.1 carrier
Build expression plasmid pcDNA3.1-BMP-2.
Step 3. the preparation of BMP-2 modified polysaccharide-graphene quantum dot gel when, after rinsing be added mannitol freeze again
It is dry.
3. middle BMP-2 and polysaccharide-graphene quantum dot gel mass ratio are 1:2.5~20 to step.
The present invention has the effect of positive: (1) Bone targeting medicine-carried system of the invention is polysaccharide-graphite of BMP-2 modification
Alkene quantum dot gel composite, in the compound, BMP-2 is target head, after being connected with hydrophilic carrier, increases BMP-2's
Hydrophily and effective local concentration can obtain better clinical efficacy.
Bone targeting medicine-carried system of the invention not only possesses what its persursor material (polysaccharide-graphene quantum dot) had
The excellent performance of three-dimensional netted stereochemical structure, large specific surface area, medicament slow release, and tests prove that should after BMP-2 modification
Gel medicine-carried system Bone targeting, and Bone targeting selectivity is high.Therefore medicine-carried system of the invention is have Bone targeting excellent
Pharmaceutical carrier.
(2) in Bone targeting medicine-carried system of the invention, the not instead of BMP-2 utilized promotes the performance of cell differentiation, sharp
Use its as growth factor have bone affinity performance, therefore in Bone targeting medicine-carried system of the invention BMP-2 as target head
Rather than active constituent.
(3) preparation method of the invention realizes polysaccharide-graphene quantum dot gel purpose of BMP-2 modification, and makes
Preparation Method is simple, is suitble to industrialized production.
Detailed description of the invention
Fig. 1 is the graph of relation of the gel bulk concentration and adsorption rate according to the preparation of 1 method of embodiment；
Fig. 2 is the graph of relation of the gel bulk concentration and adsorbance according to the preparation of 1 method of embodiment；
Fig. 3 is the graph of relation of the gel bulk concentration and adsorption rate according to the preparation of 4 method of embodiment；
Fig. 4 is the graph of relation of the gel bulk concentration and adsorbance according to the preparation of 4 method of embodiment；
Fig. 5 is the graph of relation of the gel bulk concentration and adsorption rate according to the preparation of 5 method of embodiment；
Fig. 6 is the graph of relation of the gel bulk concentration and adsorbance according to the preparation of 5 method of embodiment；
Fig. 7 is the graph of relation of the gel bulk concentration and adsorption rate according to the preparation of 6 method of embodiment；
Fig. 8 is the graph of relation of the gel bulk concentration and adsorbance according to the preparation of 6 method of embodiment.
Specific embodiment
(embodiment 1)
The Bone targeting medicine-carried system of the present embodiment is chitosan-graphene quantum dot gel composite of BMP-2 modification, BMP-2
It is fairlead or target head, chitosan-graphene quantum dot gel is pharmaceutical carrier.Adhere to as fairlead or the BMP-2 of target head
In chitosan-graphene quantum dot gel surface.
The preparation method of the Bone targeting medicine-carried system of the present embodiment the following steps are included:
1. preparing chitosan-graphene quantum dot gel.
2 g citric acid powders are added in crucible, are placed in program-controlled cabinet-type electric furnace in 100 DEG C~300 DEG C (the present embodiment
In be 200 DEG C) under calcine 30~50 min(the present embodiment in be 30min).After calcined product is cooled to room temperature, take out, accurately
Weigh and to be made into concentration be in 1~4mg/mL(the present embodiment is 2mg/mL with ultrapure water dissolution) graphene quantum dot solution,
After completely dissolution to product, filter graphene quantum dot yellow transparent solution, be kept in dark place.
2.5g Chitosan powder is dissolved in be configured in 1~4mg/mL(the present embodiment in 0.1 mol/L acetum be
The graphene quantum dot solution of aforementioned preparation is added in 2mg/mL) chitosan solution thereto under stiring, continues after being added dropwise
It is stirred to react 10min；10%NaOH solution to yellow solid precipitate object is added dropwise into reacting rear material to be precipitated, sediment is filtered,
Washing three times, and is freeze-dried 24 h at -45 DEG C, obtains chitosan-graphene quantum dot gel.
2. the building of BMP-2 plasmid.
Using the genomic DNA extracted in rat peripheral blood, BMP-2 gene is synthesized by polymerase chain reaction (PCR),
It is subjected to genetic recombination with pcDNA3.1 carrier, transformed competence colibacillus Escherichia coli construct expression plasmid pcDNA3.1-BMP-2.
3. BMP-2 modifies the preparation of chitosan-graphene quantum dot gel.
By step, 1. 2. the middle BMP-2 plasmid prepared soaks for the middle chitosan-graphene quantum dot gel prepared and step
Extract is centrifuged after mixing, is placed in water-bath, and after 50 DEG C of water-baths preheat 20min, vortex is mixed, then it is placed at room temperature for 30 ±
Chitosan-graphene quantum dot-BMP-2 CDNA is obtained within 10 minutes, 30min is centrifuged under 10000r/min revolving speed, abandons supernatant
After distilled water rinses 3 times, 1% mannitol is added in liquid, be freeze-dried at -40 DEG C obtain afterwards for 24 hours BMP-2 modification shell it is poly-
Sugar-graphene quantum dot gel.Wherein BMP-2 and chitosan-graphene quantum dot gel mass ratio are 1 ︰ 2.5~20, this
It is 1 ︰ 10 in embodiment.
In addition to 1 ︰ 10 that the present embodiment enumerates, BMP-2 can also be with chitosan-graphene quantum dot gel mass ratio
Other ratios in 1 ︰, 2.5~20 ranges such as 1 ︰ 2.5,1 ︰ 4,1 ︰ 15,1 ︰ 20.
(embodiment 2)
Remaining is same as Example 1 for the preparation method of the Bone targeting medicine-carried system of the present embodiment, the difference is that:
Step 3. in, after chitosan-graphene quantum dot gel and BMP-2 plasmid leaching liquor mix, preheated in 40 DEG C of water-baths
30min。
(embodiment 3)
Remaining is same as Example 1 for the preparation method of the Bone targeting medicine-carried system of the present embodiment, the difference is that:
Step 3. in, after chitosan-graphene quantum dot gel and BMP-2 plasmid leaching liquor mix, preheated in 60 DEG C of water-baths
10min。
(embodiment 4)
The Bone targeting medicine-carried system of the present embodiment is sodium alginate-graphene quantum dot gel composite of BMP-2 modification, BMP-
2 be fairlead or target head, and sodium alginate-graphene quantum dot gel is pharmaceutical carrier.BMP-2 as fairlead or target head is attached
In sodium alginate-graphene quantum dot gel surface.
The preparation method of the Bone targeting medicine-carried system of the present embodiment the following steps are included:
1. preparing sodium alginate-graphene quantum dot gel.
2 g citric acid powders are added in crucible, are placed in program-controlled cabinet-type electric furnace in 100 DEG C~300 DEG C (the present embodiment
In be 200 DEG C) under calcine 30~50 min(the present embodiment in be 30min).After calcined product is cooled to room temperature, take out, accurately
Weigh and to be made into concentration be in 1~4mg/mL(the present embodiment is 2mg/mL with ultrapure water dissolution) graphene quantum dot solution,
After completely dissolution to product, filter graphene quantum dot yellow transparent solution, be kept in dark place.
Weigh 0.2g sodium alginate, be dissolved in 10mL distilled water, at 60 DEG C heating stirring to forming uniform liquid (3h),
The graphene quantum dot solution of aforementioned preparation is added thereto under stiring, continues to be stirred to react after being added dropwise uniform to being formed
10%NaOH solution to solid sediment is added dropwise into reacting rear material and is precipitated, sediment is filtered, washed three for suspension (5h)
It is secondary, and 24 h are freeze-dried at -45 DEG C, obtain sodium alginate-graphene quantum dot gel.
2. the building of BMP-2 plasmid.Method is the same as embodiment 1.
3. BMP-2 modifies the preparation of sodium alginate-graphene quantum dot gel.
By step, 1. 2. the middle BMP-2 plasmid prepared is made for the middle sodium alginate-graphene quantum dot gel prepared and step
Leaching liquor is centrifuged after mixing, is placed in water-bath, and after 60 DEG C of water-baths preheat 30min, vortex is mixed, then is placed at room temperature for 30
Sodium alginate-graphene quantum dot-BMP-2 CDNA is obtained within ± 10 minutes, is centrifuged 30min under 10000r/min revolving speed, in abandoning
Clear liquid after distilled water rinses 3 times, is added 1% mannitol, the seaweed for obtaining BMP-2 modification afterwards for 24 hours is freeze-dried at -40 DEG C
Sour sodium-graphene quantum dot gel.Wherein BMP-2 and sodium alginate-graphene quantum dot gel mass ratio be 1 ︰ 2.5~
20, it is 1 ︰ 8 in the present embodiment.
In addition to 1 ︰ 8 that the present embodiment enumerates, the mass ratio of BMP-2 and chitin-sodium alginate gel can also be 1 ︰
2.5, other ratios in 1 ︰, 2.5~20 range such as 1 ︰ 4,1 ︰ 10,1 ︰ 20.
(embodiment 5)
The Bone targeting medicine-carried system of the present embodiment is lactobionic acid-graphene quantum dot gel composite of BMP-2 modification, BMP-2
It is fairlead or target head, lactobionic acid-graphene quantum dot gel is pharmaceutical carrier.Adhere to as fairlead or the BMP-2 of target head
In lactobionic acid-graphene quantum dot gel surface.
The preparation method of the Bone targeting medicine-carried system of the present embodiment the following steps are included:
1. preparing lactobionic acid-graphene quantum dot gel.
2 g citric acid powders are added in crucible, is placed in program-controlled cabinet-type electric furnace and calcines 50 min at 300 DEG C.It forges
It after burning product is cooled to room temperature, takes out, accurately weigh and is made into the graphene quantum dot that concentration is 4mg/mL with ultrapure water dissolution
Solution, after completely dissolution to product, filter graphene quantum dot yellow transparent solution, be kept in dark place.
40mg lactobionic acid is weighed, (pH=6) are dissolved in 20mL phosphate buffer), aforementioned match is added thereto under stiring
The graphene quantum dot solution of system continues to be stirred to react for 24 hours after being added dropwise, and 10%NaOH solution is added dropwise into reacting rear material
It is precipitated to solid sediment, sediment is filtered, washed three times, and be freeze-dried 24 h at -45 DEG C, obtains lactobionic acid-stone
Black alkene quantum dot gel.
2. the building of BMP-2 plasmid.Method is the same as embodiment 1.
3. BMP-2 modifies the preparation of lactobionic acid-graphene quantum dot gel.Method is with embodiment 1, wherein BMP-2 and cream
Saccharic acid-graphene quantum dot gel mass ratio is 1 ︰ 2.5~20, is 1 ︰ 10 in the present embodiment.
In addition to 1 ︰ 10 that the present embodiment enumerates, BMP-2 can also be with lactobionic acid-graphene quantum dot gel mass ratio
Other ratios in 1 ︰, 2.5~20 ranges such as 1 ︰ 2.5,1 ︰ 4,1 ︰ 15,1 ︰ 20.
(embodiment 6)
The Bone targeting medicine-carried system of the present embodiment is beta-cyclodextrin-graphene quantum dot gel composite of BMP-2 modification, BMP-
2 be fairlead or target head, and beta-cyclodextrin-graphene quantum dot gel is pharmaceutical carrier.BMP-2 as fairlead or target head is attached
In beta-cyclodextrin-graphene quantum dot gel surface.
The preparation method of the Bone targeting medicine-carried system of the present embodiment the following steps are included:
1. preparing beta-cyclodextrin-graphene quantum dot gel.
2 g citric acid powders are added in crucible, being placed in program-controlled cabinet-type electric furnace in is to calcine 35min at 200 DEG C.
After calcined product is cooled to room temperature, take out, accurately weigh and with ultrapure water dissolution be made into concentration be apply be in example 2mg/mL stone
Black alkene quantum dot solution, after completely dissolution to product, filter graphene quantum dot yellow transparent solution, be kept in dark place.
Beta-cyclodextrin, polypropylene, citric acid are dissolved in the n,N-Dimethylformamide of 10 mL, N, N- dimethyl formyl
Beta-cyclodextrin concentration 15% in amine mixed solution, polypropylene concentration 3%, citric acid concentration 3%；Magnetic agitation 5h is heated to 60 DEG C,
The graphene quantum dot solution for adding aforementioned preparation continues to be stirred to react for 24 hours after being added dropwise, be added dropwise into reacting rear material
10%NaOH solution to solid sediment is precipitated, and sediment is filtered, washed three times, and be freeze-dried 24 h at -45 DEG C, is obtained
To beta-cyclodextrin-graphene quantum dot gel.
2. the building of BMP-2 plasmid.Method is the same as embodiment 1.
3. BMP-2 modifies the preparation of beta-cyclodextrin-graphene quantum dot gel.Method with embodiment 1, wherein BMP-2 with
Beta-cyclodextrin-graphene quantum dot gel mass ratio is 1 ︰ 2.5~20, is 1 ︰ 12 in the present embodiment.
In addition to 1 ︰ 12 that the present embodiment enumerates, BMP-2 and beta-cyclodextrin-graphene quantum dot gel mass ratio can be with
It is other ratios in 1 ︰, 2.5~20 ranges such as 1 ︰ 2.5,1 ︰ 4,1 ︰ 15,1 ︰ 20.
(test example, polysaccharide-graphene quantum dot gel targeting of BMP-2 modification are tested)
Test method: polysaccharide-graphene of the BMP-2 modification of polysaccharide-graphene quantum dot gel and 3 samples is prepared respectively
Quantum dot gel.
In polysaccharide-graphene quantum dot gel of the BMP-2 modification of 3 samples, polysaccharide-graphene quantum dot gel and bone
The mass ratio for targeting fairlead BMP-2 is 10:1,4:1,2.5:1.
This four parts of gelinites are diluted into 10 times, 15 times, 20 times respectively, it is spare.
Each two parts of gelinite after taking 10mL to dilute, portion adsorbs preceding control, a addition hydroxyapatite 40mg, and two
After part shakes 10min, constant temperature places 3h at 37 DEG C, and 12000rpm is centrifuged 15min, and supernatant is taken to survey absorbance at 208nm
A。
Take hydroxyapatite 40mg, distilled water 10mL be added, after shaking 10min, 37 DEG C of constant temperature place 3h, 12000rpm from
Heart 15min takes supernatant to correct as the blank group of absorption group, and control group is using distilled water as blank before adsorbing.
Test result is as follows:
(1) for the BMP-2 of the chitosan-graphene quantum dot gel and 3 samples that prepare according to the method for embodiment 1
The relation curve of the chitosan of modification-graphene quantum dot gel, gel bulk concentration and adsorption rate is shown in Fig. 1, gel bulk concentration with
The relation curve of adsorbance is shown in Fig. 2.
The chitosan that equivalent hydroxyapatite modifies various concentration Bone targeting BMP-2-graphene quantum dot gelinite
Absorption result is shown in Fig. 1 and Fig. 2.
By Fig. 1 and Fig. 2 it is found that chitosan-graphene quantum dot gel is to the adsorption rate of hydroxyapatite only 6% or so, and
Adsorption rate up to 80% of the chitosan-graphene quantum dot gelinite of BMP-2 modification to hydroxyapatite.
Adsorption rate and suction of the 40mg hydroxyapatite to Bone targeting gelinite in 1000ug/mL ~ 3000ug/mL concentration range
There are certain regularities for attached amount, and with the raising of concentration, adsorption rate has downward trend, and adsorbance increases.Rather than Bone targeting rouge
Plastid changes with concentration, and certain regularity is not present to the adsorption rate variation tendency of hydroxyapatite.
(2) sodium alginate-graphene quantum dot gel for the method preparation according to embodiment 4 and 3 samples
The relation curve of sodium alginate-graphene quantum dot gel of BMP-2 modification, gel bulk concentration and adsorption rate is shown in Fig. 3, gelinite
The relation curve of concentration and adsorbance is shown in Fig. 4.
As can be seen from figs. 3 and 4 sodium alginate-graphene quantum dot gel is to the adsorption rate of hydroxyapatite only 6% or so,
And sodium alginate-graphene quantum dot gelinite of BMP-2 modification is to the adsorption rate up to 80% of hydroxyapatite.
Adsorption rate and suction of the 40mg hydroxyapatite to Bone targeting gelinite in 1000ug/mL ~ 3000ug/mL concentration range
There are certain regularities for attached amount, and with the raising of concentration, adsorption rate has downward trend, and adsorbance increases.Rather than Bone targeting rouge
Plastid changes with concentration, and certain regularity is not present to the adsorption rate variation tendency of hydroxyapatite.
(3) lactobionic acid-graphene quantum dot gel for the method preparation according to embodiment 5 and 3 samples
The relation curve of lactobionic acid-graphene quantum dot gel of BMP-2 modification, gel bulk concentration and adsorption rate is shown in that Fig. 5, gelinite are dense
Degree and the relation curve of adsorbance are shown in Fig. 6.
By Fig. 5 and Fig. 6 it is found that lactobionic acid-graphene quantum dot gel is to the adsorption rate of hydroxyapatite only 6% or so, and
Adsorption rate up to 86% of the lactobionic acid-graphene quantum dot gelinite of BMP-2 modification to hydroxyapatite.
Adsorption rate and suction of the 40mg hydroxyapatite to Bone targeting gelinite in 1000ug/mL ~ 3000ug/mL concentration range
There are certain regularities for attached amount, and with the raising of concentration, adsorption rate has downward trend, and adsorbance increases.Rather than Bone targeting rouge
Plastid changes with concentration, and certain regularity is not present to the adsorption rate variation tendency of hydroxyapatite.
(4) beta-cyclodextrin-graphene quantum dot gel for the method preparation according to embodiment 6 and 3 samples
The relation curve of beta-cyclodextrin-graphene quantum dot gel of BMP-2 modification, gel bulk concentration and adsorption rate is shown in Fig. 7, gelinite
The relation curve of concentration and adsorbance is shown in Fig. 8.
By Fig. 7 and Fig. 8 it is found that beta-cyclodextrin-graphene quantum dot gel is to the adsorption rate of hydroxyapatite only 6% or so,
And beta-cyclodextrin-graphene quantum dot gelinite of BMP-2 modification is to the adsorption rate up to 85% of hydroxyapatite.
Adsorption rate and suction of the 40mg hydroxyapatite to Bone targeting gelinite in 1000ug/mL ~ 3000ug/mL concentration range
There are certain regularities for attached amount, and with the raising of concentration, adsorption rate has downward trend, and adsorbance increases.Rather than Bone targeting rouge
Plastid changes with concentration, and certain regularity is not present to the adsorption rate variation tendency of hydroxyapatite.
The above various embodiments and application examples are the explanations to a specific embodiment of the invention, rather than to limit of the invention
System, person skilled in the relevant technique without departing from the spirit and scope of the present invention, can also make various changes
Change and change and obtain corresponding equivalent technical solution, thus all equivalent technical solutions should be included into it is of the invention
Scope of patent protection.
Claims (7)
1. a kind of Bone targeting medicine-carried system, it is characterised in that: it is polysaccharide-graphene quantum dot gel composite of BMP-2 modification,
BMP-2 is fairlead or target head, and polysaccharide-graphene quantum dot gel is pharmaceutical carrier；BMP-2 as fairlead or target head is attached
In polysaccharide-graphene quantum dot gel surface.
2. Bone targeting medicine-carried system according to claim 1, it is characterised in that: the polysaccharide be chitosan, sodium alginate,
One of lactobionic acid, beta-cyclodextrin.
3. Bone targeting medicine-carried system according to claim 1, it is characterised in that: BMP-2 and polysaccharide-graphene quantum dot are solidifying
The mass ratio of glue is 1:2.5~20.
4. the preparation method of Bone targeting medicine-carried system according to claim 1, it is characterised in that the following steps are included:
1. preparing polysaccharide-graphene quantum dot gel；
2. the building of BMP-2 plasmid；
3. the preparation of BMP-2 modified polysaccharide-graphene quantum dot gel: by step 1. in prepare polysaccharide-graphene quantum dot
2. the middle BMP-2 plasmid leaching liquor centrifugation prepared after mixing, is placed in water-bath, in 40 DEG C~60 DEG C water for gel and step
After 10~30min of bath preheating, vortex is mixed, then is placed at room temperature for, and obtains polysaccharide-graphene quantum dot-BMP-2 CDNA, is centrifuged,
Supernatant is abandoned, obtains chitosan-graphene quantum dot gel of BMP-2 modification after rinsing, freeze-drying.
5. the preparation method of Bone targeting medicine-carried system according to claim 4, it is characterised in that:
Step 2. the building of BMP-2 plasmid when, using the genomic DNA extracted in rat peripheral blood, pass through polymerase chain reaction
It answers (PCR) to synthesize BMP-2 gene, it is subjected to genetic recombination with pcDNA3.1 carrier, transformed competence colibacillus Escherichia coli construct table
Up to plasmid pcDNA3.1-BMP-2.
6. the preparation method of Bone targeting medicine-carried system according to claim 4, it is characterised in that: 3. BMP-2 is modified step
Mannitol is added when the preparation of polysaccharide-graphene quantum dot gel, after rinsing to be freeze-dried again.
7. the preparation method of Bone targeting medicine-carried system according to claim 4, it is characterised in that: step 3. in BMP-2 with
Polysaccharide-graphene quantum dot gel mass ratio is 1:2.5~20.
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| CN103100114A (en) * | 2013-01-23 | 2013-05-15 | 西南交通大学 | Preparation method of medical metal surface slow-released growth factor coating |
| CN105477647A (en) * | 2015-12-02 | 2016-04-13 | 常州大学 | Graphene quantum dot/chitosan xerogel preparation and application of graphene quantum dot/chitosan xerogel to fluorescent imaging and drug sustained release |
| KR20170090358A (en) * | 2016-01-28 | 2017-08-07 | 경희대학교 산학협력단 | Organic-inorganic hybrid micro particles and use therof |
| CN107176996A (en) * | 2017-05-15 | 2017-09-19 | 中国人民解放军第三军医大学第附属医院 | Nucleic acid and recombinant plasmid and its preparation method and application |
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|---|---|---|---|---|
| CN103100114A (en) * | 2013-01-23 | 2013-05-15 | 西南交通大学 | Preparation method of medical metal surface slow-released growth factor coating |
| CN105477647A (en) * | 2015-12-02 | 2016-04-13 | 常州大学 | Graphene quantum dot/chitosan xerogel preparation and application of graphene quantum dot/chitosan xerogel to fluorescent imaging and drug sustained release |
| KR20170090358A (en) * | 2016-01-28 | 2017-08-07 | 경희대학교 산학협력단 | Organic-inorganic hybrid micro particles and use therof |
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