CN104524589B - The magnetic corpusculum pharmaceutical carriers of double crosslinker AMB 1, its preparation method and application - Google Patents
The magnetic corpusculum pharmaceutical carriers of double crosslinker AMB 1, its preparation method and application Download PDFInfo
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- CN104524589B CN104524589B CN201410748312.0A CN201410748312A CN104524589B CN 104524589 B CN104524589 B CN 104524589B CN 201410748312 A CN201410748312 A CN 201410748312A CN 104524589 B CN104524589 B CN 104524589B
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Abstract
The invention discloses a kind of magnetic corpusculum pharmaceutical carriers of double crosslinker AMB 1, its preparation method and application, using the magnetic corpusculums of AMB 1 with natural lipid bimolecular film layer as carrier, using Geniposide and poly- L glutamic acid as bimolecular cross-linked material, the magnetic corpusculum pharmaceutical carriers of double crosslinker AMB 1 are formed, crosslinking single medicine or multi-medicament can be carried and targeting drug delivery system is built.The present invention uses natural double crosslinker, and toxicity is low, good biocompatibility, the drugloading rate and envelop rate of the obtained magnetic corpusculum pharmaceutical carriers of double crosslinker AMB 1 are improved compared with conventional art, single and multi-medicament high-efficient carrier can be achieved, no burst effect, deenergized period is long, the performance of medicine effect itself is not influenceed, the effect of drug combination, Synergistic can also be realized while target administration, and preparation technology is simple, it is easy to operate, environmental protection, has broad application prospects.
Description
Technical field
The present invention relates to a kind of double crosslinker AMB-1 magnetic corpusculums pharmaceutical carrier, its preparation method and application.
Background technology
The research of pharmaceutical carrier is always one of focus of field of medicaments, and wherein the exploitation of new material using in medicine with carrying
Critical role is occupied in body research.Magnetic corpusculum is that its diameter is tens to hundreds of by the coated single magnetic domain crystal of Lipid bilayer membranes
Between nanometer, main composition includes Fe3O4、FeS、Fe3S4Or FeS2.Magnetic corpusculum synthesized by different magnetotactic bacterias is big
It is variant in terms of small, quantity, composition and morphosis, that is, possess species specificity.One kind is derived from
Contain substantial amounts of protein and amino on the Magnetosome membrane of Magnetospirillum magneticum AMB-1 magnetotactic bacterias
Acid, these albumen and amino acid (such as primary amino radical) are in some two-way crosslinking agent (glutaraldehyde, glyoxal, two succinimide fat
Fat acid fat etc.) in the presence of can be used for linking bioactive molecule.The magnetic corpusculum AMB-1 has as the application of pharmaceutical carrier
Report, but it is many using single chemical cross-linking agent at present, and toxicity is larger, drugloading rate relatively low about 10% or so, and envelop rate is limited,
Single medicine can only be often carried, its application is received larger limitation.
The content of the invention
It is an object of the invention to overcome the deficiencies in the prior art part, there is provided a kind of double crosslinker AMB-1 magnetic corpusculums
Pharmaceutical carrier, its preparation method and application, using the AMB-1 magnetic corpusculum with natural lipid bimolecular film layer as carrier matrix material
Material, using Biological cross-linker Geniposide and L-glutamic acid as bimolecular cross-linked material, by single medicine or multi-medicament respectively with
Above two material is well mixed, and concussion reaction formation crosslinking, the targeting for forming double crosslinker AMB-1 magnetic corpusculum pharmaceutical carriers is given
Medicine system.
One of the technical solution adopted for the present invention to solve the technical problems is：
A kind of double crosslinker AMB-1 magnetic corpusculum pharmaceutical carrier, the double crosslinker is Geniposide and L-glutamic acid.
In one embodiment：The AMB-1 magnetic corpusculum derives from Magnetospirillum magneticum AMB-1 magnetotactics
Bacterium.
The two of the technical solution adopted for the present invention to solve the technical problems are：
A kind of preparation method of double crosslinker AMB-1 magnetic corpusculum pharmaceutical carrier, weighs AMB-1 magnetic corpusculum particles, by its point
Dissipate in pH 7.3~7.5 PBS solution, the 0.9~1.1mg/mL of quality such as addition and AMB-1 magnetic corpusculum particles poly- L- paddy
Propylhomoserin solution, 45~55W ultrasonic disperses, 4~6min is fully to mix, and the concentration that adding Geniposide makes it in mixed solution is
0.04%~0.85%；Continue 45~55W, 0.9~1.1min of ultrasonic disperse, be spaced 4~6min, repeat ultrasound 8~12 times, will
Mixed solution after ultrasound is placed in 35~39 DEG C, 10~75h is crosslinked in 50~70rpm shaking table, obtains described double crosslinker
AMB-1 magnetic corpusculum pharmaceutical carriers.
The three of the technical solution adopted for the present invention to solve the technical problems are：
A kind of double crosslinker AMB-1 magnetic corpusculum pharmaceutical carriers are building the application on targeting drug delivery system.
The four of the technical solution adopted for the present invention to solve the technical problems are：
A kind of targeting drug delivery system of double crosslinker AMB-1 magnetic corpusculum pharmaceutical carrier, the double crosslinker AMB-1 magnetic corpusculums
Carried on pharmaceutical carrier for medicine of the one or more containing amino group.
In one embodiment：Carried on the double crosslinker AMB-1 magnetic corpusculum pharmaceutical carrier for cytarabine, daunorubicin
One or both of combination.
The five of the technical solution adopted for the present invention to solve the technical problems are：
A kind of preparation method of the targeting drug delivery system of double crosslinker AMB-1 magnetic corpusculum pharmaceutical carrier, takes double crosslinker
In AMB-1 magnetic corpusculum pharmaceutical carriers, the PBS solution for being scattered in pH 7.3~7.5, addition and AMB-1 magnetic corpusculum particles etc.
0.9~1.1mg/mL solution of the carrying medicine of quality, 45~55W, 0.9~1.1min of ultrasonic disperse are spaced 4~6min, repeated
Ultrasound 8~12 times, is placed in 35~39 DEG C by the mixed solution after ultrasound, 10~75h is crosslinked in 50~70rpm shaking table, obtain
The targeting drug delivery system of described double crosslinker AMB-1 magnetic corpusculum pharmaceutical carriers.
In one embodiment：It is described to carry the combination that medicine is one or both of cytarabine, daunorubicin.
In one embodiment：In 0.9~1.1mg/mL solution of the carrying medicine of the quality such as addition and AMB-1 magnetic corpusculum particles
While add crosslinking agent.
The technical program is compared with background technology, and it has the following advantages that：
1.AMB-1 magnetic corpusculum derives from Magnetospirillum magneticum AMB-1 magnetotactic bacterias, with natural
External lipid bilayer and internal mineral crystal (Fe3O4Or Fe3S4) structure, it is a kind of natural targeting vector；Capital
Buddhist nun puts down extracts obtained natural biological crosslinking agent to be a kind of from cape jasmine fruit, and its toxicity is far below glutaraldehyde and many other normal
With crosslinking agent, while there is significant curative effect to digestive system, cardiovascular system and central nervous system disease, also resist with certain
Scorching and treatment soft tissue injury effect；L-glutamic acid strand contains a large amount of carboxyls, and degradable is glutamic acid, is 20 kinds of human body
One of amino acid；Therefore, using Geniposide mediation crosslinking L-glutamic acid in double crosslinker AMB-1 made from Magnetosome membrane surface
Magnetic corpusculum pharmaceutical carrier possesses good biocompatibility, the medical/medicinal efficacy also aided in, in biomedicine, pharmacy, life
The fields such as thing material have broad application prospects.
2. double crosslinker AMB-1 magnetic corpusculum pharmaceutical carrier produced by the present invention has, form is homogeneous, particle diameter distribution is narrower, raw
The features such as thing film parcel and good biocompatibility, and pass through a large amount of carboxyls of L-glutamic acid strand introducing, drugloading rate and encapsulating
Rate is improved compared with conventional art, and single and multi-medicament high-efficient carrier can be achieved, and no burst effect, deenergized period is long, does not influence
The performance of medicine effect itself, can also realize the effect of drug combination, Synergistic, with practical while target administration
And wide application prospect.
3. the preparation technology of the present invention is simple, easy to operate, poisonous organic solvent is not used, and environmental protection meets and worked as
The requirement of preceding environmentally friendly section bar material.
Embodiment
Present disclosure is illustrated below by embodiment：
Embodiment 1
Accurate weighing is purified and AMB-1 magnetic corpusculum particles after being freeze-dried, and the AMB-1 magnetic corpusculums derive from
In Magnetospirillum magneticum AMB-1 magnetotactic bacterias, the PBS solution for being scattered in pH 7.4, add with
The 1mg/mL of quality such as AMB-1 magnetic corpusculum particles poly-L-glutamic acid acid solution, 50W ultrasonic disperses 5min is added with fully mixing
Geniposide makes its concentration in mixed solution be 0.5%；Continue 50W ultrasonic disperse 1min, be spaced 5min, repeat ultrasound 10
It is secondary, the mixed solution after ultrasound is placed in 37 DEG C, 72h is crosslinked in 60rpm shaking table, double crosslinker AMB-1 magnetic corpusculum medicines are obtained
Thing carrier.
Embodiment 2
Accurate weighing is purified and AMB-1 magnetic corpusculum particles after being freeze-dried, and the AMB-1 magnetic corpusculums derive from
In Magnetospirillum magneticum AMB-1 magnetotactic bacterias, the PBS solution for being scattered in pH 7.4, add with
The 1mg/mL of quality such as AMB-1 magnetic corpusculum particles poly-L-glutamic acid acid solution, 50W ultrasonic disperses 5min is added with fully mixing
Geniposide makes its concentration in mixed solution be 0.05%；Continue 50W ultrasonic disperse 1min, be spaced 5min, repeat ultrasound 10
It is secondary, the mixed solution after ultrasound is placed in 37 DEG C, 48h is crosslinked in 60rpm shaking table, double crosslinker AMB-1 magnetic corpusculum medicines are obtained
Thing carrier.
Embodiment 3
Accurate weighing is purified and AMB-1 magnetic corpusculum particles after being freeze-dried, and the AMB-1 magnetic corpusculums derive from
In Magnetospirillum magneticum AMB-1 magnetotactic bacterias, the PBS solution for being scattered in pH 7.4, add with
The 1mg/mL of quality such as AMB-1 magnetic corpusculum particles poly-L-glutamic acid acid solution, 50W ultrasonic disperses 5min is added with fully mixing
Geniposide makes its concentration in mixed solution be 0.1%；Continue 50W ultrasonic disperse 1min, be spaced 5min, repeat ultrasound 10
It is secondary, the mixed solution after ultrasound is placed in 37 DEG C, 12h is crosslinked in 60rpm shaking table, double crosslinker AMB-1 magnetic corpusculum medicines are obtained
Thing carrier.
Embodiment 4
Accurate weighing is purified and AMB-1 magnetic corpusculum particles after being freeze-dried, and the AMB-1 magnetic corpusculums derive from
In Magnetospirillum magneticum AMB-1 magnetotactic bacterias, the PBS solution for being scattered in pH 7.4, add with
The 1mg/mL of quality such as AMB-1 magnetic corpusculum particles poly-L-glutamic acid acid solution, 50W ultrasonic disperses 5min is added with fully mixing
Geniposide makes its concentration in mixed solution be 0.8%；Continue 50W ultrasonic disperse 1min, be spaced 5min, repeat ultrasound 10
It is secondary, the mixed solution after ultrasound is placed in 37 DEG C, 24h is crosslinked in 60rpm shaking table, double crosslinker AMB-1 magnetic corpusculum medicines are obtained
Thing carrier.
The preparation of the cytarabine targeting drug delivery system (ABMs-P) of embodiment 5
The double crosslinker AMB-1 magnetic corpusculum pharmaceutical carriers obtained in Example 1, the PBS for being scattered in pH 7.4 is molten
In liquid, the 1mg/mL solution and crosslinking agent 1- (3- dimethylaminos third of the cytarabine of quality such as addition and AMB-1 magnetic corpusculum particles
Base) -3- ethyl carbodiimide hydrochloride solution, the addition of crosslinking agent is this area routine techniques, 50W ultrasonic disperse 1min,
Every 5min, ultrasound is repeated 10 times, the mixed solution after ultrasound is placed in 37 DEG C, 24h is crosslinked in 60rpm shaking table, ABMs- is obtained
P。
After testing, the drugloading rate and envelop rate of cytarabine respectively reach 38.9 ± 2.4% and 64.1 ± 6.6%；Through releasing
Performance is put, cytarabine effectively can discharge from ABMs-P, in the medicine of sustainable release nearly 90% in 40 days；Suppression
Knurl test result indicates that, ABMs-P and cytarabine bulk drug show obvious inhibitory action to HL-60 cells.The targeting
Delivery system is successfully constructed, it was demonstrated that introduce molecular bridge in Magnetosome membrane surface by Geniposide mediation crosslinking L-glutamic acid
Beam, can increase effective binding site of Magnetosome membrane surface drug, to realize that the combined loading of multi-medicament provides basis.
The preparation of the daunorubicin targeting drug delivery system of embodiment 6
The double crosslinker AMB-1 magnetic corpusculum pharmaceutical carriers obtained in Example 2, the PBS for being scattered in pH 7.4 is molten
In liquid, the 1mg/mL solution and crosslinking agent 1- (3- dimethylaminos third of the daunorubicin of quality such as addition and AMB-1 magnetic corpusculum particles
Base) -3- ethyl carbodiimide hydrochloride solution, the addition of crosslinking agent is this area routine techniques, 50W ultrasonic disperse 1min,
Every 5min, ultrasound is repeated 10 times, the mixed solution after ultrasound is placed in 37 DEG C, 12h is crosslinked in 60rpm shaking table, obtains soft red
Mycin targeting drug delivery system.
Embodiment 7 carries the preparation of cytarabine/daunorubicin targeting drug delivery system (ADBMs-P) altogether
The double crosslinker AMB-1 magnetic corpusculum pharmaceutical carriers obtained in Example 1, the PBS for being scattered in pH 7.4 is molten
In liquid, the matter such as the 1mg/mL solution of the cytarabine of quality such as addition and AMB-1 magnetic corpusculum particles and AMB-1 magnetic corpusculum particles
1mg/mL solution and crosslinking agent 1- (3- dimethylamino-propyls) -3- ethyl carbodiimide hydrochloride solution of the daunorubicin of amount, is handed over
The addition for joining agent is this area routine techniques, and 50W ultrasonic disperse 1min are spaced 5min, repeat ultrasound 10 times, after ultrasound
Mixed solution is placed in 37 DEG C, is crosslinked 48h in 60rpm shaking table, obtains ADBMs-P.
After testing, the drugloading rate and envelop rate of cytarabine respectively reach 34.1% ± 1.0% and 70.6% ± 3.9%,
Daunorubicin then obtains 17.3% ± 1.8% and 36.1% ± 5.4% drugloading rate and envelop rate respectively；Examined through release performance
Examine, cytarabine can effectively discharge with daunorubicin from ADBMs-P, and without phenomenon of burst release is produced, cytarabine is in 40
Sustained release 85% in it, daunorubicin is then in almost release completely in 13 days；In addition, inhibiting tumor assay result shows ADBMs-P
The increment of HL-60 cells can effectively be suppressed, its tumor killing effect is similar to bulk drug.It can thus be concluded that, this carries magnetic target administration altogether
System is successfully constructed, it was demonstrated that magnetic corpusculum load multi-medicament will likely should as a kind of novel therapeutic mode of great potential
Effective treatment for a variety of diseases.
Embodiment 8 carries the preparation of cytarabine/daunorubicin targeting drug delivery system (ADBMs-P) altogether
The double crosslinker AMB-1 magnetic corpusculum pharmaceutical carriers obtained in Example 3, the PBS for being scattered in pH 7.4 is molten
In liquid, the matter such as the 1mg/mL solution of the cytarabine of quality such as addition and AMB-1 magnetic corpusculum particles and AMB-1 magnetic corpusculum particles
1mg/mL solution and crosslinking agent 1- (3- dimethylamino-propyls) -3- ethyl carbodiimide hydrochloride solution of the daunorubicin of amount, is handed over
The addition for joining agent is this area routine techniques, and 50W ultrasonic disperse 1min are spaced 5min, repeat ultrasound 10 times, after ultrasound
Mixed solution is placed in 37 DEG C, is crosslinked 72h in 60rpm shaking table, obtains ADBMs-P.
It is described above, only present pre-ferred embodiments, therefore the scope that the present invention is implemented can not be limited according to this, i.e., according to
The equivalent changes and modifications that the scope of the claims of the present invention and description are made, all should still belong in the range of the present invention covers.
Claims (7)
1. a kind of double crosslinker AMB-1 magnetic corpusculum pharmaceutical carrier, it is characterised in that：The double crosslinker is Geniposide and poly- L- paddy
Propylhomoserin；The AMB-1 magnetic corpusculum derives from Magnetospirillum magneticum AMB-1 magnetotactic bacterias；The double cross
Connection agent AMB-1 magnetic corpusculum pharmaceutical carriers are prepared by following preparation method：AMB-1 magnetic corpusculum particles are weighed, are scattered in
In pH 7.3~7.5 PBS solution, the 0.9~1.1mg/mL of quality such as addition and AMB-1 magnetic corpusculum particles L-glutamic acid
Solution, 45~55W ultrasonic disperses, 4~6min is fully to mix, and the concentration that adding Geniposide makes it in mixed solution is
0.04%~0.85%；Continue 45~55W, 0.9~1.1min of ultrasonic disperse, be spaced 4~6min, repeat ultrasound 8~12 times, will
Mixed solution after ultrasound is placed in 35~39 DEG C, 10~75h is crosslinked in 50~70rpm shaking table, obtains described double crosslinker
AMB-1 magnetic corpusculum pharmaceutical carriers.
2. a kind of double crosslinker AMB-1 magnetic corpusculum pharmaceutical carriers according to claim 1 are being built on targeting drug delivery system
Application.
3. a kind of targeting drug delivery system of double crosslinker AMB-1 magnetic corpusculum pharmaceutical carrier according to claim 1, its feature
It is：Carried on the double crosslinker AMB-1 magnetic corpusculum pharmaceutical carrier for medicine of the one or more containing amino group.
4. a kind of targeting drug delivery system of double crosslinker AMB-1 magnetic corpusculum pharmaceutical carrier according to claim 3, its feature
It is：Carried on the double crosslinker AMB-1 magnetic corpusculum pharmaceutical carrier for one or both of cytarabine, daunorubicin
Combination.
5. a kind of preparation of the targeting drug delivery system of double crosslinker AMB-1 magnetic corpusculum pharmaceutical carrier according to claim 4
Method, it is characterised in that：Double crosslinker AMB-1 magnetic corpusculum pharmaceutical carriers are taken, pH 7.3~7.5 PBS solution is scattered in
In, 0.9~1.1mg/mL solution of the carrying medicine of the quality such as addition and AMB-1 magnetic corpusculum particles, 45~55W ultrasonic disperses
0.9~1.1min, be spaced 4~6min, repeat ultrasound 8~12 times, by the mixed solution after ultrasound be placed in 35~39 DEG C, 50~
10~75h is crosslinked in 70rpm shaking table, the targeting drug delivery system of described double crosslinker AMB-1 magnetic corpusculum pharmaceutical carriers is obtained.
6. a kind of preparation of the targeting drug delivery system of double crosslinker AMB-1 magnetic corpusculum pharmaceutical carrier according to claim 5
Method, it is characterised in that：It is described to carry the combination that medicine is one or both of cytarabine, daunorubicin.
7. a kind of preparation of the targeting drug delivery system of double crosslinker AMB-1 magnetic corpusculum pharmaceutical carrier according to claim 5
Method, it is characterised in that：In 0.9~1.1mg/mL solution of the carrying medicines of the quality such as addition and AMB-1 magnetic corpusculum particles
Crosslinking agent is added simultaneously.
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Citations (2)
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| CN1927400A (en) * | 2006-09-25 | 2007-03-14 | 唐喜庆 | Biomagnetism nano target anti-cancer drug and its preparation |
| CN102989005A (en) * | 2012-12-05 | 2013-03-27 | 华侨大学 | Methotrexate-loaded magnetosome drug carrier and preparation method thereof |
-
2014
- 2014-12-09 CN CN201410748312.0A patent/CN104524589B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1927400A (en) * | 2006-09-25 | 2007-03-14 | 唐喜庆 | Biomagnetism nano target anti-cancer drug and its preparation |
| CN102989005A (en) * | 2012-12-05 | 2013-03-27 | 华侨大学 | Methotrexate-loaded magnetosome drug carrier and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| 《FacilefabricationofhighperformancesMTXnanocompositeswithnatural biomembrane bacterialnanoparticlesusing GP》;YuangangLiu et al.;《Materials Letters》;20130321;第100卷;摘要，249页右栏第2段 * |
| 《天然磁靶向纳米药物载体磁小体研究进展》;邓琼嘉;《科学通报》;20140420;第59卷(第11期);948页左栏 * |
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