CN100460016C - Meso-porous nanometer particle of calcium phosphate, its preparation method and application - Google Patents
Meso-porous nanometer particle of calcium phosphate, its preparation method and application Download PDFInfo
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- CN100460016C CN100460016C CNB2006100351107A CN200610035110A CN100460016C CN 100460016 C CN100460016 C CN 100460016C CN B2006100351107 A CNB2006100351107 A CN B2006100351107A CN 200610035110 A CN200610035110 A CN 200610035110A CN 100460016 C CN100460016 C CN 100460016C
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Abstract
The invention dis closes a nanomater granule of mesoporous calcium phosphate which is used as medicine carrier. It comprises Ca10(PO4)6(OH)2, the crystalline is nanometer banded shape, the width of band is 40-150 nm, length is 5-10 um, specific surface area is 160-280 m2.g-1 and the acverage bore diameter is 3-8 nm. The said prepartion process comprises following steps: adding phosphate radical solution, calcium ion solution in order into solution containing surface active agent and cosurfactant, getting mixing solution; dripping alkaline liquor slowly and stirring at the same into mixing solution to regulate pH to 8-11; stirring for reaction for 1-48 hours and getting reaction product of white deposition; washing white deposition with organic dissolvent and water and getting nanomater granule of mesoporous calcium phosphate. The product is characterized by no toxicity, good biocompatibility, high medicine carrying amount and a certain degradability.
Description
Technical field
The present invention relates to a kind of nano material, particularly a kind of meso-porous nanometer particle of calcium phosphate.
The invention still further relates to a kind of preparation method of above-mentioned nano-particle.
The invention still further relates to the application of described meso-porous nanometer particle of calcium phosphate
Background technology
Pharmaceutical carrier is a kind of biologic material products that rises gradually with the development of biomaterial science, clinical medicine and materia medica.Medicine enters human body by carrier, and carrier makes drug release position, speed close mode etc. to absorption, parcel and the bonding of medicine to have selectivity and controllability, realize the slow release and the targeting transmission of medicine, thereby better bring into play medication effect.Developing rapidly of nanometer biotechnology is for its application at biomedicine field provides important enlightenment and opportunity.Utilize nanotechnology to make the pharmaceutical carrier nanorize, can make the drug-carrying nanometer particle particle diameter little, specific surface area is big, adhesion to receptor tissue is big, anelasticity reaches with time of contact, the contact area organized and all greatly increases after the administration, thereby can improve bioavailability of medicament, reduction toxicity, minimizing dosing etc.
Existing in recent years both at home and abroad many patents of nano-medicament carrier technology of preparing and bibliographical information mainly are organic polymer class materials.Patent: as the preparation method (world) of paracyanogen base NBCA nano-particle, the patent No.: PCT/CN2004/001089; The preparation method (world) of galactosyl-hsa magnetic adriamycin nanoparticle, the patent No.: PCT/CN2004/001091; The preparation method of amycin-paracyanogen base NBCA nanoparticle, the patent No.: 200410046648.9; The preparation method of albumin nano granular, the patent No.: 200410046677.5; The preparation method of glycosyl galactose hydroxyapatite-poly-D-lysine, the patent No.: 200410046678.X; The preparation method of paracyanogen base NBCA nanoparticle, the patent No.: 200410046649.3; The preparation method of galactose albumin magnetic adriamycin nanoparticle, the patent No.: 200410046650.6.Bibliographical information: high molecular polymer is handed over fat～glycolide copolymerization (PLCA) as third, particle diameter 70～160nm, J.Control.Release.1998.54.201-211 such as () Song.C; Polylactic acid (PLA), particle diameter 123 ± 23nm (J.Control.Release.2000.65.221-229 such as Fishbein.I); The PLGA/PLA/PCL complex, 110-208nm, (Eur.J.Pharm.Biopharm.1998.46.137-143 such as Verger.ML-L); Paracyanogen base acrylic acid alkyl fat (PACA), 120～230nm (Biochem.Phamacol.2000.59.105-114 such as Ranc.D.F), and solid lipid nanoparticle (SLN) pharmaceutical carrier such as camptothecine～stearic acid SLN, particle diameter 197nm (.Pharm.Res.1999.16.751-757 such as Yang.S); Ciclosporin A～stearic acid SLN, particle diameter 316nm (Int.J.Pharm.2000.200.153-159 such as Zhang.Q).
Than general material, the medicine carrier material must have excellent biological compatibility, absorbability, nontoxic and do not have a savings.The character of material is for the release of medicine and be absorbed with and significant effects, and studies show that organic polymer class material has following defective as pharmaceutical carrier: dissolubility at first in vivo is lower, and influence absorbs; And the protein in the blood is adsorbed onto carrier surface easily, causes the prominent of medicine to release; Also can have been reduced the medicament contg of other tissue in addition, and hydrophilic inorganic material can reduce prominent releasing, and be difficult for the identification of coverlet nuclear phagocyte by reticuloendothelial system phagocytic.In addition, it is nonpolar that Organic substance mostly is, and water solublity is low, and monomer, few monomer, surfactant require numerous and diverse purge process in the preparation, and the inorganic drug carrier can be avoided above-mentioned shortcoming.Therefore, the inorganic nano pharmaceutical carrier also receives publicity in recent years.As nano red elemental selenium, and particle diameter 20～60nm (Gao Xueyun etc., Chinese public health, 2000.5.421-424); Magnetic nano particle Fe
3O
4, (Alexiou.C etc., Cancer.Research.2000.6641-6648; Lubbe.A.S etc., Cancer.Research.1996.56.4694-4701; Zhang Yangde etc., the contemporary Chinese medical journal, 2001.3.14-16), nanometer SiO2, (Ahola.M, Biomaterials.2001.22.2163-2170); Mesoporous nano SiO2 MCM-41, (Chem.Mater such as Vallet.R.M., 2001.13.308-311) etc.It is relatively poor that but the defective of all these inorganic nano pharmaceutical carriers is its biocompatibility and degradability, has only by metabolism and could get rid of, and takes for a long time and might accumulate in vivo.If therefore can synthesize locate a kind of nontoxic, drug loading is high, can and energy biodegradable inorganic nano material compatible with human-body biological, just can overcome above-mentioned drawback with it as pharmaceutical carrier.And the nano-calcium phosphate first-selection of this material just.
Calcium phosphate has and similar composition of mineralization of skeleton thing and similar structure, calcium phosphate mainly is among form with hydroxyapatite nano level acicular crystal is distributed in collagenous network along an orientation in people's natural bone, belong to bioactive materials, nontoxic, have good biocompatibility.Its biodegradation is in proper order: amorphous calcium phosphate〉calcium hydrogen phosphate〉phosphoric acid oxygen four calcium〉type alpha tricalcium phosphate〉bata-tricalcium phosphate〉hydroxyapatite.Under the effect of body fluid, slowly dissolving and ionizing take place in amorphous nano-calcium phosphate, can slowly discharge medicine.Therefore employing as pharmaceutical carrier, utilizes its different dissolubility without the nano-calcium phosphate of crystallization degree, controls medicine rate of release in vivo, can reach the purpose of long-acting release.
Domestic and international at present the synthetic and rarely seen report of medicine carrying about meso-porous nano calcium phosphate.The most frequently used method of preparation mesoporous material is the method by the molecule self assembly.The molecule assembling is by weak covalency, non-covalent bond---hydrogen bond, ionic bond (electrostatic interaction), and hydrophobic interaction, the hydrogen bond of Van der Waals force and water produces.Although these keys are obviously isolated comparatively speaking, they are as an entire combination together the time, but controlling the formation of macromolecular structure and intermolecular interaction (Zhang.S.G., naturebiotechnology.2003.10.1171-1178).People can control these intermolecular combinations, and (Reches.M is etc., Scence.2003.300.625-637 to construct different shape and pattern nano material; Sang.O.K, etc., Nature.2003.424.411-414), and represent fabulous application prospect.As utilize the duct medicine carrying (Chen.J.F. etc. of mesoporous silicon oxide, Biomaterials, 2004.25.723-727), the functionalization appearance bonding drug molecule of one dimension CNT (Dwain.F.E, etc., ExpertOpin.Biol.Ther.2003.3.655-663.) etc.Though the medicine carrying of mesoporous silicon oxide and CNT is good, their biological degradability and biological activity are relatively poor.
Summary of the invention
The objective of the invention is to overcome the defective of prior art, a kind of nontoxic, good biocompatibility is provided, drug loading is high and has had the meso-porous nanometer particle of calcium phosphate of certain degradability, useful as drug carrier.
Another object of the present invention is to provide a kind of preparation method of described nano-particle.
Meso-porous nanometer particle of calcium phosphate of the present invention contains Ca
10(PO
4)
6(OH)
2Component, the crystal shape of described nano-particle becomes nano strip, bandwidth is 40～150nm, length is 5～10 μ m, specific surface area is 160～280m
2G
-1, average pore size is 3～8nm.
The preparation method of meso-porous nanometer particle of calcium phosphate of the present invention comprises the steps:
(1). in the aqueous solution that contains surfactant and cosurfactant, add PO successively
4 3-Solion, Ca
2+Solion makes Ca
2+Ionic molar content is 0.3～1.2molL
-1, PO
4 3-Ionic molar content is 0.5～1.5molL
-1, the molar content of surfactant is 0.2～1.0molL
-1, the molar content of cosurfactant is 0.2～1.0molL
-1
(2). slowly dripping concentration in above-mentioned solution while stirring is 1～5molL
-1Alkali liquor, regulate its pH value to 8～11, be under 80～150 ℃ in temperature, continued stirring reaction 1～48 hour, obtaining product is white precipitate, with organic solvent and water washing white precipitate, promptly obtains meso-porous nanometer particle of calcium phosphate of the present invention.
Described surfactant is a kind of or more than one mixture in octadecyl phosphoric acid fat, cetyl phosphoric acid fat, dodecylphosphoric acid fat, the eight alkyl phosphoric acid fat; Described cosurfactant is a kind of or more than one mixture in n-butyl alcohol, ethanol, the ethylene glycol.
Described aqueous slkali is the general alkaline solution in this area, a kind of or more than one mixture of the preferred NaOH of the present invention, KOH, ammonia spirit.
Described organic solvent can adopt the general organic solvent in this area, a kind of or more than one mixture in the preferred methyl ether of the present invention, methanol, ethanol, the acetone.
Meso-porous nanometer particle of calcium phosphate of the present invention, as pharmaceutical carrier, it is big to have a drug loading, the characteristics that envelop rate is high.As get 3g poly--ε-oneself in be dissolved in 10ml CH
2C
12, add mass ratio (rhodamine B medicine: polymer: meso-porous nanometer particle of calcium phosphate) be the medicine of 1:1:1, the meso-porous nanometer particle of calcium phosphate of rhodamine B and surface hydrophilic ball milling in water, lyophilizing then respectively.Its China and foreign countries' water is 300ml, and emulsifying agent adopts methylcellulose 600mg (0.2w/v%), its drug loading 30.62%, envelop rate 91.86%.
The present invention compared with prior art has following advantage:
1. nano-particle of the present invention is biocompatibility, degradability inorganic nano carrier, and is nontoxic, harmless.
2. nanoparticulate dispersed of the present invention is good, has bigger draw ratio, and specific surface area is big, and hole is evenly distributed, and helps the loading of medicine, the drug loading height.
3. method process equipment of the present invention is simple, and medicine source is abundant, and is cheap, helps suitability for industrialized production.
Description of drawings
Fig. 1 is the transmission electron microscope figure of the meso-porous nanometer particle of calcium phosphate of embodiment 1 preparation, and wherein: Fig. 1 (a) is the transmission electron microscope figure of low range, and Fig. 1 (b) is powerful transmission electron microscope figure;
Fig. 2 is the X-x ray diffration pattern x of the meso-porous nanometer particle of calcium phosphate of embodiment 1 preparation, and wherein: Fig. 2 (a) is low-angle X-x ray diffration pattern x, and Fig. 2 (b) is the X-x ray diffration pattern x of wide-angle;
Fig. 3 is the Fourier transform infrared spectroscopy figure of the meso-porous nanometer particle of calcium phosphate of embodiment 1 preparation;
Fig. 4 is the graph of pore diameter distribution of the meso-porous nanometer particle of calcium phosphate of embodiment 1 preparation;
Fig. 5 is a medicine carrying microballoons enlarged photograph of making preparing carriers of meso-porous nanometer particle of calcium phosphate.
The specific embodiment
Following embodiment further specifies of the present invention, is not limitation of the present invention.
The dodecylphosphoric acid liposoluble that takes by weighing 14g adds 15ml0.5molL successively in the 30ml deionized water and in the alcoholic acid mixed solution of 30ml
-1Ca
2+Solution and 15ml0.3molL
-1PO
4 3-Solution behind the mix homogeneously, is used 2molL
-1NaOH solution adjust pH=10, then 90 ℃ down the evaporation reflow treatment obtained white precipitate in 24 hours, reuse methanol wash product obtains mesoporous calcium phosphate nano powder body after the drying.Wherein the molar concentration of above-mentioned each component is:
Ca
2+Ion 0.5molL
-1, PO
4 3-Ion 0.3molL
-1, dodecylphosphoric acid fat 0.88molL
-1
The TEM photo shows that granule is nano strip, and bandwidth is 40～100nm, and length is 5～10 μ m (Fig. 1 (a), Fig. 1 (b)), and the duct is evenly distributed, and helps medicine carrying.Specific surface area is 160m
2G
-1, average pore size is 3.5nm (Fig. 4), pore volume is 0.21cm
3G
-1Its low angle of XRD test shows is mesoporous characteristic peak (Fig. 2 (a)), and crystalline phase is mainly weak crystalline hydroxyapatite Ca
10(PO
4)
6(OH)
2Component (Fig. 2 (b)) also contains the phosphate of more amorphous state simultaneously, helps degraded.The FTIR spectrum analysis shows that band is at 1000～1100cm
-1Between be three-fold degeneracy v
3, P-O antisymmetric stretching vibration mould; 961cm
-1About band be v
1, nondegenerate P-O symmetrical stretching vibration mould; Band is at 573cm
-1Between be three-fold degeneracy v
4, the O-P-O bending mode; Band is at 471cm
-1Between be double degenerate v
2, the O-P-O bending mode.Hydroxyl translational vibration v
1(OH) be 3571cm
-1, (pendulum) dynamic model v shakes
L(OH-1) 631cm
-1, be hydroxyapatite characteristic peak (Fig. 3).
The medicine carrying microballoons of doing preparing carriers with this meso-porous nanometer particle of calcium phosphate as shown in Figure 5.
The cetyl phosphoric acid liposoluble that takes by weighing 12g adds 10ml0.5molL successively in the 20ml deionized water and in the alcoholic acid mixed solution of 20ml
-1Ca
2-Solution and 10ml0.3molL
-1PO
4 3-Solution behind the mix homogeneously, is used 3molL
-1NaOH solution adjust pH=8, then 100 ℃ down the evaporation reflow treatment obtained white precipitate in 12 hours, reuse methanol wash product obtains mesoporous calcium phosphate nano powder body after the drying.Wherein the molar concentration of above-mentioned each component is:
Ca
2+Ion 0.5molL
-1, PO
4 3-Ion 0.3molL
-1, cetyl phosphoric acid fat 0.93molL
-1
Resulting mesoporous calcium phosphate nano powder body specific surface area is 210m
2G
-1, pore volume is 0.23cm
3G
-1, average pore size is 5.2nm.
The TEM photo shows that granule is nano strip, and its bandwidth is 90～120nm, and length reaches 6～8 μ m, and the duct is evenly distributed, and helps medicine carrying.Large-angle X-ray diffraction shows that the principal crystalline phase of the mesoporous calcium phosphate that this system obtains is weak crystalline hydroxyapatite, also contains the phosphate of more amorphous state simultaneously, helps degraded.
The octadecyl phosphoric acid liposoluble that takes by weighing 16g adds 12ml0.5molL successively in the 25ml deionized water and in the alcoholic acid mixed solution of 25ml
-1Ca
2+Solution and 12ml0.3molL
-1PO
4 3-Solution behind the mix homogeneously, is used 4molL
-1NaOH solution adjust pH=9, then 110 ℃ down the evaporation reflow treatment obtained white precipitate in 36 hours, reuse methanol wash product obtains mesoporous calcium phosphate nano powder body after the drying.Wherein the molar concentration of above-mentioned each component is:
Ca
2+Ion 0.5molL
-1, PO
4 3-Ion 0.3molL
-1, octadecyl phosphoric acid fat 0.91molL
-1
Resulting mesoporous calcium phosphate nano powder body specific surface area is 240m
2G
-1, pore volume is 0.25cm
3G
-1, average pore size is 7.5nm.
The TEM photo shows that granule is nano strip, and its bandwidth is 100～120nm, and length reaches 8～7 μ m, and the duct is evenly distributed, and helps medicine carrying.Large-angle X-ray diffraction shows that the principal crystalline phase of the mesoporous calcium phosphate that this system obtains is weak crystalline hydroxyapatite, also contains the phosphate of more amorphous state simultaneously, helps degraded.
The eight alkyl phosphoric acid liposoluble that take by weighing 10g add 10ml0.5molL successively in the 20ml deionized water and in the alcoholic acid mixed solution of 20ml
-1Ca
2+Solution and 10ml0.3molL
-1PO
4 3-Solution behind the mix homogeneously, is used 4.5molL
-1NaOH solution adjust pH=11, then 130 ℃ down the evaporation reflow treatment obtained white precipitate in 40 hours, reuse methanol wash product obtains mesoporous calcium phosphate nano powder body after the drying.Wherein the molar concentration of above-mentioned each component is:
Ca
2+Ion 0.5molL
-1, PO
4 3-Ion 0.3molL, eight alkyl phosphoric acid fat 1.2molL
-1
Resulting mesoporous calcium phosphate nano powder body specific surface area is 100m
2G
-1, pore volume is 0.18cm
3G
-1, average pore size is 3.0nm.
The TEM photo shows that granule is nano strip, and its bandwidth is 70～100nm, and length reaches 7～10 μ m, and the duct is evenly distributed, and helps medicine carrying.Large-angle X-ray diffraction shows that the principal crystalline phase of the mesoporous calcium phosphate that this system obtains is weak crystalline hydroxyapatite, also contains the phosphate of more amorphous state simultaneously, helps degraded.
Claims (3)
1. the preparation method of a meso-porous nanometer particle of calcium phosphate is characterized in that comprising the steps:
(1) in the aqueous solution that contains surfactant and cosurfactant, adds PO successively
4 3-Solion, Ca
2+Solion makes Ca
2+Ionic molar content is 0.3～1.2molL
-1, PO
4 3-Ionic molar content is 0.5～1.5molL
-1, the molar content of surfactant is 0.2～1.0molL
-1, the molar content of cosurfactant is 0.2～1.0molL
-1
(2) slowly dripping concentration while stirring in above-mentioned solution is 1～5molL
-1Alkali liquor, regulate its pH value to 8～11, be under 80～150 ℃ in temperature, continued stirring reaction 1～48 hour, obtaining product is white precipitate, with organic solvent and water washing white precipitate, promptly obtains meso-porous nanometer particle of calcium phosphate;
Surfactant described in the step (1) is a kind of or more than one mixture in octadecyl phosphoric acid fat, cetyl phosphoric acid fat, dodecylphosphoric acid fat, the eight alkyl phosphoric acid fat;
Cosurfactant described in the step (1) is a kind of or more than one mixture in n-butyl alcohol, ethanol, the ethylene glycol.
2. method according to claim 1 is characterized in that alkali liquor described in the step (2) is a kind of of NaOH, KOH, ammonia spirit or more than one mixture.
3. method according to claim 1 and 2 is characterized in that the organic solvent described in the step (2) is a kind of or more than one mixture in methyl ether, methanol, ethanol, the acetone.
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Families Citing this family (13)
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|---|---|---|---|---|
| MY162982A (en) * | 2008-09-23 | 2017-07-31 | Laboratory Skin Care Inc | Active agent loaded uniform, rigid, spherical, nanoporous calcium phosphate particles and methods of making and using the same |
| TWI556839B (en) * | 2009-05-06 | 2016-11-11 | 研究室護膚股份有限公司 | Dermal delivery compositions comprising active agent-calcium phosphate particle complexes and methods of using the same |
| CN105919945A (en) * | 2009-09-23 | 2016-09-07 | 实验室护肤股份有限公司 | Activator-loaded uniform, rigid, spherical and nano-porous calcium phosphate particles as well as preparing and using methods thereof |
| WO2011081874A1 (en) | 2009-12-29 | 2011-07-07 | W. R. Grace & Co.-Conn. | Composite inorganic particles and methods of making and using the same |
| CN101979091B (en) * | 2010-11-04 | 2012-01-25 | 中国科学院上海硅酸盐研究所 | Preparation method of calcium phosphate nano medicament-carrying systems |
| CN102100927B (en) * | 2011-01-22 | 2014-04-16 | 浙江理工大学 | Porous hydroxy calcium phosphate nanometer particle-modified titanium-based titanate nanowire biologic support material and preparation method thereof |
| CN103142477B (en) * | 2013-04-08 | 2015-09-30 | 薛巍 | A kind of preparation method containing nano-carrier microemulsion formulation of sustained-release colchicine micro emulsion |
| CN104445130B (en) * | 2014-11-27 | 2016-09-14 | 武汉理工大学 | A kind of mesoporous bata-tricalcium phosphate raw powder's production technology |
| CN104587488B (en) * | 2015-02-02 | 2017-10-03 | 哈尔滨工业大学 | A kind of preparation method of the mesoporous apatite nano-medicament carrier to liver cancer cells with pH responses and cell-targeting |
| CN104548108B (en) * | 2015-02-02 | 2017-06-20 | 哈尔滨工业大学 | A kind of preparation method of the mesoporous apatite nano-medicament carrier of pH responses core shell structure |
| CN111115598B (en) * | 2018-10-30 | 2023-06-13 | 天津工业大学 | Method for synthesizing multilevel structure hydroxyapatite by ketone guidance |
| CN114804059B (en) * | 2022-03-09 | 2024-02-27 | 深圳市华科创智技术有限公司 | Fluorapatite quasi-equiaxial nanocrystalline and preparation method thereof |
| CN115636400B (en) * | 2022-11-04 | 2023-12-05 | 山东大学 | Preparation method of one-dimensional multifunctional hydroxyapatite nano-belt with secondary structure and application of nano-belt in assembling functional stem cell spheres |
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