CN110170057A - A kind of nanometer grain preparation method of the double medicines of the load of Tf modification and its application - Google Patents
A kind of nanometer grain preparation method of the double medicines of the load of Tf modification and its application Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
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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
- A61K31/65—Tetracyclines
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
- A61K47/6927—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores
- A61K47/6929—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle
- A61K47/6931—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer
- A61K47/6935—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer the polymer being obtained otherwise than by reactions involving carbon to carbon unsaturated bonds, e.g. polyesters, polyamides or polyglycerol
- A61K47/6937—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer the polymer being obtained otherwise than by reactions involving carbon to carbon unsaturated bonds, e.g. polyesters, polyamides or polyglycerol the polymer being PLGA, PLA or polyglycolic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
Abstract
The present invention relates to pharmaceutical technology fields, more particularly to a kind of nanometer grain preparation method of the double medicines of load of transferrins (Tf) modification, the following steps are included: PEG-PLGA amphiphilic polymer, hydrophobic drug and hydrophilic medicament are mixed, lotion one is obtained；By lotion one and emulsifier mixing and emulsifying, lotion two is obtained, the volume ratio of lotion one and emulsifier is 1: 5；The nano particle loaded after being separated to lotion two；It uses Tf as the ligand of nano particle, using PEG-PLGA amphiphilic polymer as framework material, is loaded using emulsifier volatility process, obtain the nano particle of the double medicines of load of Tf modification.The beneficial effects of the present invention are: the present invention selects and has that toxicity is low, hydrophily and the preferable PEG-PLGA amphiphilic polymer of biological degradability are as nano-medicament carrier, good biocompatibility, non-immunogenicity is highly-safe；And the hydrophilic radical of PEG can reduce blood to the opsonic action of nanoparticle, to extend the blood halflife of drug.
Description
Technical field
The present invention relates to pharmaceutical technology fields, and in particular to a kind of nanometer grain preparation method of the double medicines of load of Tf modification
And its application.
Background technique
After cerebral ischemia certain time restores blood supply, function not only fails to restore, and more serious brain function but occurs
Obstacle, referred to as Cerebral Ischemia/Reperfusion (CIR) damage, mechanism is complex, is mainly excessively formed with free radical, excitability ammonia
Base acid toxicity, the number of mechanisms such as super calcium, inflammatory reaction are related into the cell.The mark of inflammatory reaction be leucocyte infiltration and small glue
The activation of cell plastid, astroglia.Inflammatory cell can release cytotoxin, including active oxygen, inflammatory factor and add
Weight cerebral lesion, the inflammatory reaction of microglia and ischemia-reperfusion are closely related, it is believed that are that potential macrophage is thin in brain
Born of the same parents: microglia is activated in a few minutes after ischemic, at the same increase interleukin-11 β, can lead with tumor necrosis factor α
Microglia is caused further to activate.So passing research has been found that nerve cell, endothelial cell and microglia
It is played an important role in ischemia-reperfusion, inhibits the damage of nerve cell and endothelial cell, while inhibiting microglia
Activation, can achieve synergistic effect, mitigates the damage of ischemia-reperfusion.
Butylphenyl phthaleine has good protective effect for Ischemic Stroke, equal for the protection of neuroprotection and endothelial cell
There is good effect, cerebral injury caused by ischemic can be mitigated by number of mechanisms, comprising: improves microcirculation, antiplatelet, resists
Oxygen, it is anti-oxidant the effects of.Minocycline is a kind of antibiotic of Tetracyclines, but in recent years, more document shows minot
Ring element has good inhibiting effect for neuroinflamation, and there are also documents to show that minocycline can inhibit enterobacteriaceae, improves intestines
Road microenvironment significantly increases the scope of application of minocycline, finds in the research of neuroinflamation, and minocycline can press down
The activation of microglia processed, effect and the activation of p38MAPK, oxidative damage, PKC etc. are related, also there is positive evidence table
Bright, in ischemia-reperfusion injury model, minocycline can play neuroprotection.In view of butylphenyl phthaleine, minocycline in ischemic
Effect in Reperfu- sion, the two have different pharmacological actions, can be from more targets such as nerve cell, endothelial cell, spongiocyte
Point plays a role, the protection cerebral ischemia re-pouring of collaboration.So butylphenyl phthaleine and minocycline are prepared as double loads by applicant's design
The nano particle of medicine, while being modified by transferrins (Tf), since Tf can be combined with TfR in nervous centralis, play target
Afferent effect, at the same nanoparticle have well it is fat-soluble, can penetrate blood-brain barrier, so this pair load medicine nanoparticle
Theoretically there is the synergistic protective effect of ischemia-reperfusion, while having both good brain targeting, for ischemia-reperfusion
Drug research has certain positive effect.
Summary of the invention
The purpose of the present invention is to provide a kind of nanometer grain preparation method of the double medicines of load of Tf modification and its application, with
Solve the problems mentioned above in the background art.
To achieve the above object, the invention provides the following technical scheme:
A kind of nanometer grain preparation method of the double medicines of load of Tf modification, comprising the following steps:
(1) PEG-PLGA amphiphilic polymer, hydrophobic drug and hydrophilic medicament are mixed, obtains lotion one；
(2) by lotion one and emulsifier mixing and emulsifying, lotion two is obtained, the volume ratio of lotion one and emulsifier is 1: (5-
20)；
(3) nano particle loaded after being separated to lotion two；
(4) use Tf as the ligand of nano particle, using PEG-PLGA amphiphilic polymer as framework material, using emulsification
Agent volatility process is loaded, and the nano particle of the double medicines of load of Tf modification is obtained.
As further technical solution of the present invention are as follows: the end of the PLGA in the PEG-PLGA amphiphilic polymer is
Carboxyl or hydroxyl, molecular weight is in 10000-30000；The end of PEG is amino and maleic amide.
As further technical solution of the invention are as follows: the Tf is polypeptide, and nano particle diameter range is 100-
300nm；The hydrophobic drug is butylphenyl phthaleine, and hydrophilic medicament is minocycline hydrochloride；The amphipathic polymerization of PEG-PLGA
Object, hydrophobic drug, hydrophilic medicament mass ratio be (10-500): (1-25): (1-50)；The PEG-PLGA amphipathic
Conjunction object concentration is 10-100mg/ml, and hydrophobic drug concentration is 0.1-3.0mg/ml, and hydrophilic medicament concentration is 5-50mg/ml；
The emulsifier is one of polyvinyl alcohol, poloxamer, Tween 80, dodecyl sodium sulfate or a variety of；The emulsifier
Concentration is 1-10%.
As further technical solution of the invention are as follows: the PEG-PLGA amphiphilic polymer, hydrophobic drug, parent
The mass ratio of aqueous pharmaceutical is (100-500): (5-15): (10-25)；The PEG-PLGA amphiphilic polymer concentration is 10-
50mg/ml, hydrophobic drug concentration are 0.5-2.0mg/ml, and hydrophilic medicament concentration is 5-30mg/ml.
As further technical solution of the invention are as follows: the PEG-PLGA amphiphilic polymer concentration is 10-15mg/
Ml, hydrophobic drug concentration are 0.5-1.5mg/ml, and hydrophilic medicament concentration is 5-15mg/ml.
As further technical solution of the invention are as follows: the emulsifier is polyvinyl alcohol, poloxamer, the two volume
Than for (1-2): 1.
As further technical solution of the invention are as follows: the polyvinyl alcohol, poloxamer volume ratio be 1.5: 1.
As further technical solution of the invention are as follows: the emulsifier concentration is 1-5%.
As further technical solution of the invention are as follows: the lotion one and the volume ratio of emulsifier are 1: (2-5).
A kind of nano particle of the double medicines of the load of Tf modification is treating the application on cerebral ischemia re-pouring injured.
The beneficial effects of the present invention are: the present invention, which selects, has low toxicity, hydrophily and the preferable PEG- of biological degradability
PLGA amphiphilic polymer is as nano-medicament carrier, good biocompatibility, and non-immunogenicity is highly-safe；And the parent of PEG
Water base group can reduce blood to the opsonic action of nanoparticle, to extend the blood halflife of drug: and do not need using auxiliary
Cosolvent can generate side effect to avoid normal tissue organ, increase patient adaptability.
The present invention is amphipathic using the Tf peptide and PEG-PLGA that can specifically bind blood-brain barrier surface TfR
Polymer phase combines, and prepares Brain targeting nanoscale medicine delivery system, and that improves drug BBB penetrates efficiency, greatly improves nano-carrier
Drug crosses over the efficiency of blood-brain barrier, makes drug-rich in lesion tissue, improves the bioavilability of drug, reduce drug
Toxic side effect and effective Drug controlled release have preferable effect on treatment ischemical reperfusion injury.
Detailed description of the invention
Fig. 1 is the nano particle aspect graph of Tf of the present invention modification；
Fig. 2 is the nano particle diameter distribution map of Tf of the present invention modification；
Fig. 3 is the Zeta potential figure of the nano particle of Tf of the present invention modification；
The level of Fig. 4 is PH of the present invention when being 7.4 minocycline hydrochloride and PEG-PLGA amphiphilic polymer tablets in vitro
Comparative result figure；
The level of Fig. 5 is PH of the present invention when being 5.5 minocycline hydrochloride and PEG-PLGA amphiphilic polymer tablets in vitro
Comparative result figure；
The horizontal result pair of Fig. 6 is PH of the present invention when being 7.4 butylphenyl phthaleine and PEG-PLGA amphiphilic polymer tablets in vitro
Than figure；
The horizontal result pair of Fig. 7 is PH of the present invention when being 5.5 butylphenyl phthaleine and PEG-PLGA amphiphilic polymer tablets in vitro
Than figure；
Fig. 8 is nano particle brain tissue drug concentration level's result figure of Tf of the present invention modification.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
Embodiment 1:
A kind of nanometer grain preparation method of the double medicines of load of Tf modification, comprising the following steps:
(1) by PEG-PLGA amphiphilic polymer, hydrophobic drug and hydrophilic medicament mixed lactic, lotion one is obtained；
(2) by lotion one and emulsifier mixing and emulsifying, lotion two is obtained, the volume ratio of lotion one and emulsifier is 1: 5；
(3) nano particle loaded after being separated to lotion two；
(4) use Tf as the ligand of nano particle, using PEG-PLGA amphiphilic polymer as framework material, using emulsification
Agent volatility process is loaded, and the nano particle of the double medicines of load of Tf modification is obtained.
The end of PLGA in the PEG-PLGA amphiphilic polymer is carboxyl or hydroxyl, and molecular weight is in 10000-
30000；The end of PEG is amino and maleic amide.
The Tf is polypeptide, and nano particle diameter range is 100-300nm；The hydrophobic drug is butylphenyl phthaleine, parent
Aqueous pharmaceutical is minocycline hydrochloride；The PEG-PLGA amphiphilic polymer, hydrophobic drug, hydrophilic medicament mass ratio
It is 10: 1: 1；The PEG-PLGA amphiphilic polymer concentration is 10mg/ml, and hydrophobic drug concentration is 0.1mg/ml, hydrophilic
Property drug concentration be 5mg/ml；The emulsifier is polyvinyl alcohol, poloxamer, Tween 80, one in dodecyl sodium sulfate
Kind is a variety of；The emulsifier concentration is 1%.
The emulsifier is polyvinyl alcohol, poloxamer, and the two volume ratio is 1: 1.
The lotion one and the volume ratio of emulsifier are 1: 2.
Embodiment 2:
A kind of nanometer grain preparation method of the double medicines of load of Tf modification, comprising the following steps:
(1) by PEG-PLGA amphiphilic polymer, hydrophobic drug and hydrophilic medicament mixed lactic, lotion one is obtained:
(2) by lotion one and emulsifier mixing and emulsifying, lotion two is obtained, the volume ratio of lotion one and emulsifier is 1: 20；
(3) nano particle loaded after being separated to lotion two；
(4) use Tf as the ligand of nano particle, using PEG-PLGA amphiphilic polymer as framework material, using emulsification
Agent volatility process is loaded, and the nano particle of the double medicines of load of Tf modification is obtained.
The end of PLGA in the PEG-PLGA amphiphilic polymer is carboxyl or hydroxyl, and molecular weight is in 10000-
30000；The end of PEG is amino and maleic amide.
The Tf is polypeptide, and nano particle diameter range is 100-300nm；The hydrophobic drug is butylphenyl phthaleine, parent
Aqueous pharmaceutical is minocycline hydrochloride；The PEG-PLGA amphiphilic polymer, hydrophobic drug, hydrophilic medicament mass ratio
It is 500: 25: 50；The PEG-PLGA amphiphilic polymer concentration is 100mg/ml, and hydrophobic drug concentration is 3.0mg/ml,
Hydrophilic medicament concentration is 50mg/ml；The emulsifier is polyvinyl alcohol, in poloxamer, Tween 80, dodecyl sodium sulfate
It is one or more: the emulsifier concentration be 10%.
The emulsifier is polyvinyl alcohol, poloxamer, and the two volume ratio is 2: 1.
The lotion one and the volume ratio of emulsifier are 1: 5.
Embodiment 3:
A kind of nanometer grain preparation method of the double medicines of load of Tf modification, comprising the following steps:
(1) by PEG-PLGA amphiphilic polymer, hydrophobic drug and hydrophilic medicament mixed lactic, lotion one is obtained；
(2) by lotion one and emulsifier mixing and emulsifying, lotion two is obtained, the volume ratio of lotion one and emulsifier is 1: 2；
(3) nano particle loaded after being separated to lotion two；
(4) use Tf as the ligand of nano particle, using PEG-PLGA amphiphilic polymer as framework material, using emulsification
Agent volatility process is loaded, and the nano particle of the double medicines of load of Tf modification is obtained.
The end of PLGA in the PEG-PLGA amphiphilic polymer is carboxyl or hydroxyl, and molecular weight is in 10000-
30000；The end of PEG is amino and maleic amide.
The Tf is polypeptide, and nano particle diameter range is 100-300nm；The hydrophobic drug is butylphenyl phthaleine, parent
Aqueous pharmaceutical is minocycline hydrochloride；The PEG-PLGA amphiphilic polymer, hydrophobic drug, hydrophilic medicament mass ratio
It is 100: 5: 10；The PEG-PLGA amphiphilic polymer concentration is 10mg/ml, and hydrophobic drug concentration is 0.5mg/ml, parent
Aqueous pharmaceutical concentration is 5mg/ml.
The emulsifier is polyvinyl alcohol, poloxamer, and the two volume ratio is 1.5: 1.
The emulsifier concentration is 1%.
Embodiment 4:
A kind of nanometer grain preparation method of the double medicines of load of Tf modification, comprising the following steps:
(1) by PEG-PLGA amphiphilic polymer, hydrophobic drug and hydrophilic medicament mixed lactic, lotion one is obtained:
(2) by lotion one and emulsifier mixing and emulsifying, lotion two is obtained, the volume ratio of lotion one and emulsifier is 1: 5；
(3) nano particle loaded after being separated to lotion two；
(4) use Tf as the ligand of nano particle, using PEG-PLGA amphiphilic polymer as framework material, using emulsification
Agent volatility process is loaded, and the nano particle of the double medicines of load of Tf modification is obtained.
The end of PLGA in the PEG-PLGA amphiphilic polymer is carboxyl or hydroxyl, and molecular weight is in 10000-
30000；The end of PEG is amino and maleic amide.
The Tf is polypeptide, and nano particle diameter range is 100-300nm；The hydrophobic drug is butylphenyl phthaleine, parent
Aqueous pharmaceutical is minocycline hydrochloride；The PEG-PLGA amphiphilic polymer, hydrophobic drug, hydrophilic medicament mass ratio
It is 500: 15: 25；The PEG-PLGA amphiphilic polymer concentration is 50mg/ml, and hydrophobic drug concentration is 2.0mg/ml,
Hydrophilic medicament concentration is 30mg/ml.
The emulsifier be polyvinyl alcohol, poloxamer, the polyvinyl alcohol, poloxamer volume ratio be 1.5: 1.
The emulsifier concentration is 5%.
The lotion one and the volume ratio of emulsifier are 1: 5.
Embodiment 5:
A kind of nanometer grain preparation method of the double medicines of load of Tf modification, comprising the following steps:
(1) by PEG-PLGA amphiphilic polymer, hydrophobic drug and hydrophilic medicament mixed lactic, lotion one is obtained；
(2) by lotion one and emulsifier mixing and emulsifying, lotion two is obtained, the volume ratio of lotion one and emulsifier is 1: 3；
(3) nano particle loaded after being separated to lotion two；
(4) use Tf as the ligand of nano particle, using PEG-PLGA amphiphilic polymer as framework material, using emulsification
Agent volatility process is loaded, and the nano particle of the double medicines of load of Tf modification is obtained.
The end of PLGA in the PEG-PLGA amphiphilic polymer is carboxyl or hydroxyl, and molecular weight is in 10000-
30000；The end of PEG is amino and maleic amide.
The Tf is polypeptide, and nano particle diameter range is 100-300nm；The hydrophobic drug is butylphenyl phthaleine, parent
Aqueous pharmaceutical is minocycline hydrochloride；The PEG-PLGA amphiphilic polymer, hydrophobic drug, hydrophilic medicament mass ratio
It is 300: 10: 18；The PEG-PLGA amphiphilic polymer concentration is 15mg/ml, and hydrophobic drug concentration is 1.5mg/ml,
Hydrophilic medicament concentration is 15mg/ml.
The emulsifier is polyvinyl alcohol, poloxamer, and the two volume ratio is 1.5: 1.
The emulsifier concentration is 3%.
The lotion one and the volume ratio of emulsifier are 1: 3.
Embodiment 6:
A kind of nano particle of the double medicines of the load of Tf modification is treating the application on cerebral ischemia re-pouring injured.
Related identification and detection for the nano particle of Tf modification:
One, the detection of form:
Concentration is to dilute after the nano particle of 1.5mg/ml takes 20 μ l, and dilution dries after being added dropwise on copper mesh, then is added dropwise
2% phosphotungstic acid is dyed.Sample drying and film is formed on copper mesh after room temperature, TEM carries out morphologic observation, specifically
Form please refers to Fig. 1.
Two, the detection of partial size and Zeta potential:
Instrument detection particle diameter distribution and Zeta potential are used after preparing the nano particle dilution of 1.5mg/ml.
As a result: being in spherical, and be evenly distributed.In the detection of partial size, the average grain diameter of liposome is 98.36 ±
As a result 3.12nm is shown in Fig. 2；And the Zeta potential of liposome is -35.63 ± 4.52mV, as a result please refers to Fig. 3.
Three, the measurement result of nano particle encapsulation rate:
Using the content of minocycline hydrochloride and butylphenyl phthaleine in high performance liquid chromatography detection liposome, take nano particle mixed
Suspension 1.5ml, takes supernatant after ultracentrifugation, detection wherein free drug concentration, encapsulation rate=(1- free drug concentration/it is mixed
Drug total concentration in suspension) x100%.The detection of minocycline hydrochloride uses ODS chromatographic column (250X4.6nm, 5 μm), mobile phase
A is the distilled water of 0.1% triethylamine, and Mobile phase B is methanol, and the ratio of A and B are 70: 30, flow velocity 1ml/min, detects wave
Long 298nm, column temperature are 30 DEG C.The detection of butylphenyl phthaleine uses ODS chromatographic column (250X4.6nm, 5 μm), and mobile phase A is methanol, stream
Dynamic phase B is distilled water, and C is acetonitrile, and A: B: C ratio is 45: 45: 10, flow velocity 1ml/min, Detection wavelength 326nm, column temperature
It is 30 DEG C.
Four, nanoparticles stable is tested:
Configuring concentrations of nanoparticles is 1.0,1.5,2.0mg/ml, is respectively placed in 5,25,60 DEG C of three kinds of temperature next months,
It is sampled 1 after experiment starts, after 15,30 days, observe form and carries out the analysis of encapsulation rate, partial size, stability result looks at table
1, table 2 and table 3.
Table 1: the study on the stability (n=3) of minocycline hydrochloride encapsulation rate
Table 2: the study on the stability (n=3) of butylphenyl phthaleine encapsulation rate
The investigation (n=3) of the active Brain targeting nanoparticles stable of table 3:Tf peptide modification
Six, tablets in vitro is tested:
The butylphenyl phthaleine and 1.5mg/ml hydrochloric acid minot that concentration is the nano particle 1ml and 1.5mg/ml of 1.5mg/ml are unreal
Fast bulk pharmaceutical chemicals 1ml, is packaged in bag filter respectively, respectively in the solution of PH7.4 and 5.5, carries out tablets in vitro experiment, is putting
After sample 0.5,1,2,4,6,8,12,24,36,48h sampling, using high performance liquid chromatography send out detection medicament contg and accumulation release
Medicine rate, experimental result please refer to Fig. 4-Fig. 7.
Specific experiment case study:
In mouse Ischemia-Reperfusion Injury Model, the targeting of nano particle is detected.
Male ICR mouse 9 are chosen, ischemia-reperfusion injury model is constructed, is randomly divided into three groups, i.e. PEG-PLGA group, hydrochloric acid
Minocycline group and butylphenyl phthaleine group, respectively tail vein injection, dosage 100mg/Kg, upon administration the three of 30min, 1h, 3h
A time point randomly selects one in every group of clump respectively, then puts to death after mouse extracts brain tissue, using high performance liquid chromatography
The drug concentration of brain tissue is detected, experimental result please refers to Fig. 8.
It is obvious to a person skilled in the art that invention is not limited to the details of the above exemplary embodiments, Er Qie
In the case where without departing substantially from spirit or essential attributes of the invention, the present invention can be realized in other specific forms.Therefore, no matter
From the point of view of which point, the present embodiments are to be considered as illustrative and not restrictive, and the scope of the present invention is by appended power
Benefit requires rather than above description limits, it is intended that all by what is fallen within the meaning and scope of the equivalent elements of the claims
Variation is included within the present invention.Any reference signs in the claims should not be construed as limiting the involved claims.
In addition, it should be understood that although this specification is described in terms of embodiments, but not each embodiment is only wrapped
Containing an independent technical solution, this description of the specification is merely for the sake of clarity, and those skilled in the art should
It considers the specification as a whole, the technical solutions in the various embodiments may also be suitably combined, forms those skilled in the art
The other embodiments being understood that.
Claims (10)
1. a kind of nanometer grain preparation method of the double medicines of load of transferrins (Tf) modification, which is characterized in that including following step
It is rapid:
(1) PEG-PLGA amphiphilic polymer, hydrophobic drug and hydrophilic medicament are mixed, obtains lotion one；
(2) by lotion one and emulsifier mixing and emulsifying, lotion two is obtained, the volume ratio of lotion one and emulsifier is 1: (5-20)；
(3) nano particle loaded after being separated to lotion two；
(4) Tf is used to wave as the ligand of nano particle using PEG-PLGA amphiphilic polymer as framework material using emulsifier
Hair method is loaded, and the nano particle of the double medicines of load of Tf modification is obtained.
2. the nanometer grain preparation method of the double medicines of load of Tf modification according to claim 1, which is characterized in that described
The end of PLGA in PEG-PLGA amphiphilic polymer is carboxyl or hydroxyl, and molecular weight is in 10000-30000；The end of PEG
End is amino and maleic amide.
3. the nanometer grain preparation method of the double medicines of load of Tf modification according to claim 1, which is characterized in that the Tf
For polypeptide, nano particle diameter range is 100-300nm；The hydrophobic drug is butylphenyl phthaleine, and hydrophilic medicament is hydrochloric acid
Minocycline；The PEG-PLGA amphiphilic polymer, hydrophobic drug, hydrophilic medicament mass ratio be (10-500): (1-
25):(1-50)；The PEG-PLGA amphiphilic polymer concentration is 10-100mg/ml, and hydrophobic drug concentration is 0.1-
3.0mg/ml, hydrophilic medicament concentration are 5-50mg/ml；The emulsifier is polyvinyl alcohol, poloxamer, Tween 80,12
One of sodium alkyl sulfonate is a variety of；The emulsifier concentration is 1-10%.
4. the nanometer grain preparation method of the double medicines of load of Tf modification according to claim 3, which is characterized in that described
PEG-PLGA amphiphilic polymer, hydrophobic drug, hydrophilic medicament mass ratio be (100-500): (5-15): (10-25)；
The PEG-PLGA amphiphilic polymer concentration is 10-50mg/ml, and hydrophobic drug concentration is 0.5-2.0mg/ml, hydrophily
Drug concentration is 5-30mg/ml.
5. the nanometer grain preparation method of the double medicines of load of Tf modification according to claim 4, which is characterized in that described
PEG-PLGA amphiphilic polymer concentration is 10-15mg/ml, and hydrophobic drug concentration is 0.5-1.5mg/ml, hydrophilic medicament
Concentration is 5-15mg/ml.
6. the nanometer grain preparation method of the double medicines of load of Tf modification according to claim 3, which is characterized in that the cream
Agent is polyvinyl alcohol, poloxamer, and the two volume ratio is (1-2): 1.
7. the nanometer grain preparation method of the double medicines of load of Tf modification according to claim 6, which is characterized in that described poly-
Vinyl alcohol, poloxamer volume ratio be 1.5: 1.
8. the nanometer grain preparation method of the double medicines of load of Tf modification according to claim 3, which is characterized in that the cream
Agent concentration is 1-5%.
9. the nanometer grain preparation method of the double medicines of load of Tf modification according to claim 1, which is characterized in that the cream
The volume ratio of liquid one and emulsifier is 1: (2-5).
10. a kind of nano particle of the double medicines of load of the Tf modification as described in claim 1-9 is any is in treatment cerebral ischemia reperfusion
Application in note damage.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111529944A (en) * | 2020-04-24 | 2020-08-14 | 李付勇 | Photodynamic therapy system and method for glioma |
| CN116531524A (en) * | 2023-07-07 | 2023-08-04 | 深圳大学总医院 | Brain-targeted nanoparticle and preparation method and application thereof |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050202075A1 (en) * | 2004-03-12 | 2005-09-15 | Pardridge William M. | Delivery of genes encoding short hairpin RNA using receptor-specific nanocontainers |
| EP1859811A1 (en) * | 2006-05-27 | 2007-11-28 | Faulstich, Heinz, Dr. | Use of conjugates of amatoxins or phallotoxins with macromolecules for tumor and inflammation therapy |
| CN101732723A (en) * | 2009-12-30 | 2010-06-16 | 上海市肿瘤研究所 | Polyethylene glycol-poly(lactic-co-glycolic acid)-polylysine nano-delivery system, preparation method and application thereof |
| CN102743337A (en) * | 2011-04-20 | 2012-10-24 | 国家纳米科学中心 | Nano-particulate medicinal composition and preparation method thereof |
| CN103044686A (en) * | 2012-11-15 | 2013-04-17 | 南京工业大学 | Preparation and application of mixed structure PLGA-PLL-PEG targeting polymer carrier |
| CN103055322A (en) * | 2012-12-20 | 2013-04-24 | 华南理工大学 | Targeted sustained release medicine carrying nanoparticle and preparation method thereof |
| CN103655517A (en) * | 2013-11-19 | 2014-03-26 | 南京医科大学 | Pep-1 peptide modified gliomas targeted nano drug delivery system and preparation method thereof |
| WO2017204475A1 (en) * | 2016-05-24 | 2017-11-30 | 한양대학교 산학협력단 | Intranasal pharmaceutical composition comprising anticancer drug-containing nanoparticles for treating brain diseases |
| CN108938599A (en) * | 2018-09-25 | 2018-12-07 | 盐城工业职业技术学院 | A kind of double medicament-carried nano carriers of cancer target |
| CN109078184A (en) * | 2018-07-13 | 2018-12-25 | 国家纳米科学中心 | Load double medicine nano particles and the preparation method and application thereof |
-
2019
- 2019-04-08 CN CN201910269067.8A patent/CN110170057A/en active Pending
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050202075A1 (en) * | 2004-03-12 | 2005-09-15 | Pardridge William M. | Delivery of genes encoding short hairpin RNA using receptor-specific nanocontainers |
| EP1859811A1 (en) * | 2006-05-27 | 2007-11-28 | Faulstich, Heinz, Dr. | Use of conjugates of amatoxins or phallotoxins with macromolecules for tumor and inflammation therapy |
| CN101732723A (en) * | 2009-12-30 | 2010-06-16 | 上海市肿瘤研究所 | Polyethylene glycol-poly(lactic-co-glycolic acid)-polylysine nano-delivery system, preparation method and application thereof |
| CN102743337A (en) * | 2011-04-20 | 2012-10-24 | 国家纳米科学中心 | Nano-particulate medicinal composition and preparation method thereof |
| CN103044686A (en) * | 2012-11-15 | 2013-04-17 | 南京工业大学 | Preparation and application of mixed structure PLGA-PLL-PEG targeting polymer carrier |
| CN103055322A (en) * | 2012-12-20 | 2013-04-24 | 华南理工大学 | Targeted sustained release medicine carrying nanoparticle and preparation method thereof |
| CN103655517A (en) * | 2013-11-19 | 2014-03-26 | 南京医科大学 | Pep-1 peptide modified gliomas targeted nano drug delivery system and preparation method thereof |
| WO2017204475A1 (en) * | 2016-05-24 | 2017-11-30 | 한양대학교 산학협력단 | Intranasal pharmaceutical composition comprising anticancer drug-containing nanoparticles for treating brain diseases |
| CN109078184A (en) * | 2018-07-13 | 2018-12-25 | 国家纳米科学中心 | Load double medicine nano particles and the preparation method and application thereof |
| CN108938599A (en) * | 2018-09-25 | 2018-12-07 | 盐城工业职业技术学院 | A kind of double medicament-carried nano carriers of cancer target |
Non-Patent Citations (5)
| Title |
|---|
| GUODONG LIU等: "PEG-PLGA Nanoparticle modified by Transferrin Loading Doxorubicin: in vitro and in vivo Studies for Glioma", 《ADVANCED MATERIALS RESEARCH》 * |
| UPADHYAY PRIYANKA等: "Transferrin-decorated thymoquinone-loaded PEG-PLGA nanoparticles exhibit anticarcinogenic effect in non-small cell lung carcinoma via the modulation of miR-34a and miR-16", 《BIOMATERIALS SCIENCE》 * |
| ZHANG XINGZHEN等: "Multifunctional Polyethylene Glycol (PEG)-Poly (Lactic-Co-Glycolic Acid) (PLGA)-Based Nanoparticles Loading Doxorubicin and Tetrahydrocurcumin for Combined Chemoradiotherapy of Glioma", 《MEDICAL SCIENCE MONITOR》 * |
| 张幸真: "阿霉素和四氢姜黄素双载多功能纳米制剂的构建及其作用研究", 《山东大学硕士学位论文》 * |
| 陆彬: "《药物新剂型与新技术》", 31 July 2005, 人民卫生出版社 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111529944A (en) * | 2020-04-24 | 2020-08-14 | 李付勇 | Photodynamic therapy system and method for glioma |
| CN116531524A (en) * | 2023-07-07 | 2023-08-04 | 深圳大学总医院 | Brain-targeted nanoparticle and preparation method and application thereof |
| CN116531524B (en) * | 2023-07-07 | 2023-09-15 | 深圳大学总医院 | Brain-targeted nanoparticle and preparation method and application thereof |
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