Nothing Special   »   [go: up one dir, main page]

CN107714646A - Amphipathic nature polyalcohol micella of tumor extracellular matrix and preparation method thereof can be penetrated - Google Patents

Amphipathic nature polyalcohol micella of tumor extracellular matrix and preparation method thereof can be penetrated Download PDF

Info

Publication number
CN107714646A
CN107714646A CN201711020966.1A CN201711020966A CN107714646A CN 107714646 A CN107714646 A CN 107714646A CN 201711020966 A CN201711020966 A CN 201711020966A CN 107714646 A CN107714646 A CN 107714646A
Authority
CN
China
Prior art keywords
amphipathic nature
nature polyalcohol
collagenase
peg
extracellular matrix
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201711020966.1A
Other languages
Chinese (zh)
Inventor
曹青日
黄豪雁
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Suzhou University
Original Assignee
Suzhou University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Suzhou University filed Critical Suzhou University
Priority to CN201711020966.1A priority Critical patent/CN107714646A/en
Publication of CN107714646A publication Critical patent/CN107714646A/en
Pending legal-status Critical Current

Links

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/10Dispersions; Emulsions
    • A61K9/107Emulsions ; Emulsion preconcentrates; Micelles
    • A61K9/1075Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/335Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/337Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having four-membered rings, e.g. taxol
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/70Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/34Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/42Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Inorganic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Dispersion Chemistry (AREA)
  • Medicinal Preparation (AREA)

Abstract

The present invention relates to a kind of amphipathic nature polyalcohol micella for penetrating tumor extracellular matrix:Including antineoplastic, amphipathic nature polyalcohol and Collagenase, the antineoplastic is wrapped in inside by the amphipathic nature polyalcohol, passes through Collagenase described in covalent key connection outside the amphipathic nature polyalcohol.Present invention also offers its preparation method:Amphipathic nature polyalcohol is reacted in the presence of crosslinking agent at 20 26 DEG C in a solvent with Collagenase, removes the first organic solvent;The amphipathic nature polyalcohol for being modified with Collagenase and antineoplastic are dissolved in the second organic solvent, obtain drug solution;Drug solution is added in pure water, removes the second organic solvent, forms the amphipathic nature polyalcohol micella that can penetrate tumor extracellular matrix.The Collagenase of the micellar surface of the present invention can penetration cell epimatrix so that antineoplastic can preferably enter tumor locus, the medication amount in increase tumour cell, so as to cause the apoptosis of tumour cell.

Description

Amphipathic nature polyalcohol micella of tumor extracellular matrix and preparation method thereof can be penetrated
Technical field
The present invention relates to a kind of field of pharmaceutical preparations, more particularly to a kind of amphipathic polymerization for penetrating tumor extracellular matrix Thing micella and preparation method thereof.
Background technology
In recent years, the incidence of disease of cancer raises year by year, and serious threat the life and health of the mankind.Although treat tumour Methods and techniques it is constantly progressive, and largely improve the survival rate of patient, but be in when most of patients is fallen ill Late period, it is impossible to be treated surgically, cancer therapy drug can only be used to carry out expectant treatment.The usual poorly water-soluble of chemicals, heart The side effect such as toxicity and bone marrow suppression is more apparent, and directly application damages bigger to patient's normal cell.
Micella with PEG hydrophilic blocks can avoid drug-loading system from being identified in reticuloendothelial system by non-specificity And swallow, into by medicament transport to liver, spleen and organ in addition.Many patents and document report show, in PEG end Guidance molecular mechanism is modified, can cause medicine that there is targeting.Using transferrins as targeting factor to PLC-PEG-B nanoparticles Son carries out biological functional, can significantly improve selectively targeted ability of the PLA-PEG nano-particles in brain tumor.Although target Tropism carrier has stronger tumour-specific binding ability, but does not solve fundamentally one present in tumor microenvironment yet Serial barrier.
Extracellular matrix (extracellular matrix, ECM) is a kind of three-dimensional macromolecular network, by collagen, Fibronectin, glycoprotein, proteoglycans and glycosaminoglycan etc. form.During tumour occurs, obvious change occurs for ECM, so as to Cause the formation of fibrosing matrix to increase, rigidity increase, ECM composition over-deposits, cause abnormal ECM to remold, therefore ECM groups Branch significantly affects the delivering of medicine.
After nanoparticle is made in chemicals, curative effect can be improved and reduce toxic side effect.At present, treated using Nano medication One significant challenge of cancer is that they are difficult to penetrate tumor extracellular matrix.
The content of the invention
In order to solve the above technical problems, it is an object of the invention to provide two that one kind can penetrate tumor extracellular matrix (ECM) Parent's property polymer micelle, amphiphilic polymers micella of the invention, the ability that carrier penetrates to tumour cell, increaseization can be improved Learn medicine and enter tumour cell, so as to cause the apoptosis of tumour cell.
The invention provides the amphipathic nature polyalcohol micella that one kind can penetrate tumor extracellular matrix (ECM):Including antitumor The antineoplastic is wrapped in inside it by medicine, amphipathic nature polyalcohol and Collagenase, amphipathic nature polyalcohol, amphiphilic Property polymer outer passes through covalent key connection Collagenase.
Further, antineoplastic is the one or more in docetaxel, adriamycin and taxol.
Further, amphipathic nature polyalcohol is in PCL-PEG copolymers, PLA-PEG copolymers and PLGA-PEG copolymers One or more.Specifically, amphipathic nature polyalcohol is PCL-PEG-COOH or PLA-PEG-COOH, PLGA-PEG-COOH.Its In, PCL is polycaprolactone, and PEG is polyethylene glycol, and PLA is PLA, and PLGA is Poly(D,L-lactide-co-glycolide.
Further, the mol ratio of Collagenase and amphipathic nature polyalcohol is 1:100-1000.Preferably, collagen The mol ratio of enzyme and amphipathic nature polyalcohol is 1:100.
Further, the mass ratio of antineoplastic and amphipathic nature polyalcohol is 0.5-15:100.
Further, the molecular weight of amphipathic nature polyalcohol is 10000-25000g/mol.
Specifically, in PCL-PEG-COOH, PCL molecular weight is 8000g/mol, and PEG molecular weight is 2000g/mol; In PLA-PEG-COOH, PLA molecular weight is 8000g/mol, and PEG molecular weight is 2000g/mol;In PLGA-PEG-COOH, PLGA molecular weight is 20000g/mol, and PEG molecular weight is 2000g/mol.
Further, Collagenase is NTx protease, typeⅡ Collagen enzyme, type Ⅳ collagen protease and V One or more in collagen type enzyme.Preferably, Collagenase is type Ⅳ collagen protease, type Ⅳ collagen protease master Degrade the ECM such as type Ⅳ collagen fiber laminin, elastin laminin and nestin, can digest the collagen of denaturation.
Present invention also offers a kind of preparation method of the above-mentioned amphipathic nature polyalcohol micella for penetrating tumor extracellular matrix Comprise the following steps:
(1) amphipathic nature polyalcohol and crosslinking agent are reacted under 20-26 DEG C (room temperature) in the first organic solvent, removed Fall the first organic solvent, then reacted in pure water with Collagenase under 20-26 DEG C (room temperature), obtain being modified with collagen egg The amphipathic nature polyalcohol of white enzyme;
(2) amphipathic nature polyalcohol for being modified with Collagenase and antineoplastic are dissolved in the second organic solvent, obtained Drug solution;
(3) drug solution is added in pure water, removes the second organic solvent, form the amphiphilic that can penetrate tumor extracellular matrix Property polymer micelle.
Further, in step (1), reacted using carbodlimide method, crosslinking agent is n-hydroxysuccinimide And 1- ethyls-(3- dimethylaminopropyls) carbodiimide hydrochloride (EDC) (NHS).
Further, in step (1), the first organic solvent is removed using revolving method.
Further, in step (1), the first organic solvent is dimethyl sulfoxide, one kind or several of acetonitrile and chloroform Kind.
Further, in step (2), the second organic solvent is acetone, methanol, ethanol, dichloromethane and chloroform One or more.
Further, in step (3), drug solution is added in pure water under agitation, mixing speed 400- 700rpm, mixing time 4-8h.
In (3) step, due to containing hydrophobe segment in amphipathic nature polyalcohol, after drug solution is added in pure water, The water-wet side of polymer inside aggregation hydrophobic segment, due to antineoplastic and hydrophobic, therefore can be wrapped in external aggregation Inside is rolled in, after removing the second organic solvent, amphipathic nature polyalcohol micella can be formed.
By such scheme, the present invention at least has advantages below:
(1) by collagen enzyme modification after micellar surface, the glue by means of Collagenase to tumor extracellular matrix Former selective degradation, the therapeutic action that chemical drug is deliverrf into tumour can be effectively increased, chemotherapeutics can be increased to tumour cell Lethality.
(3) chemotherapeutics is wrapped in micelle inner core, be delivered in tumour cell, play killing tumor cell.Simultaneously The micella stability of the present invention is high, physiological compatibility is good, can increase chemicals stability, drug substance stable can be made slowly to discharge, And drug bioavailability is improved, reduce body toxic side effect.
(4) preparation technology of the invention is simple, and envelop rate is high, is easy to industrialized production;Obtained penetrates with increase Physical property of amphiphilic polymers micella of tumor extracellular matrix effect etc. meets the needs of tumour cell treatment.
Described above is only the general introduction of technical solution of the present invention, in order to better understand the technological means of the present invention, And can be practiced according to the content of specification, below with presently preferred embodiments of the present invention and coordinate accompanying drawing describe in detail as after.
Brief description of the drawings
Fig. 1 is the transmissioning electric mirror test result of the micella prepared by the embodiment of the present invention 1,2,6 and 7;
Fig. 2 is the vitro release result of micella prepared by embodiment 4.
Embodiment
With reference to the accompanying drawings and examples, the embodiment of the present invention is described in further detail.Implement below Example is used to illustrate the present invention, but is not limited to the scope of the present invention.
Except as otherwise noted, " DOX " involved in following examples represents " adriamycin ";" DTX " represents " more Xi Tasai ";" TAXOL " represents " taxol " " PEG " and represents " polyethylene glycol ";" IV collagenase " represents " IV type Collagenase ";" I collagenase " represents " type Ⅳ collagen protease ";" II collagenase " represents " II type glue Former protease ";" V collagenase " represents " V collagen type enzyme ".
In following examples, in used PCL-PEG-COOH, PCL molecular weight is 8000g/mol, PEG molecule Measure as 2000g/mol;In PLA-PEG-COOH, PLA molecular weight is 8000g/mol, and PEG molecular weight is 2000g/mol; In PLGA-PEG-COOH, PLGA molecular weight is 20000g/mol, and PEG molecular weight is 2000g/mol.
The preparation of embodiment 1.PCL-PEG-COOH blank micellas
The preparation method of the PCL-PEG blank micellas as control experiment is present embodiments provided, it is specific as follows:
Take 5mg PCL-PEG-COOH to be dissolved in 0.5mL acetone, obtain polymer solution 10mg/mL.100 μ L are above-mentioned Polymer solution and 400 μ L acetone are sufficiently mixed again.Under conditions of being stirred vigorously, it is slowly added in 5mL pure water, stirs 4h, make organic solvent evaporation.
The medicine of embodiment 2. and carrier dispensing are than being 0.5:The preparation of 100 DOX-PCL-PEG-COOH micellas
The preparation method of the DOX-PCL-PEG-COOH micellas as control experiment is present embodiments provided, it is specific as follows:
Take DOX powder 10mg molten in methyl alcohol, ultrasonic dissolution, be settled in 100mL volumetric flasks, obtain 100 μ g/mL's DOX standard solutions.Take 5mg PCL-PEG-COOH to be dissolved in 0.5mL acetone, obtain polymer solution 10mg/mL.Take 100 μ L polymer solutions and 50 μ L Doxorubicin solutions, add acetone until 500 μ L.Under conditions of being stirred vigorously, it is pure to be slowly added to 5mL In water, 4h is stirred, makes organic solvent evaporation.
The medicine of embodiment 3. and carrier dispensing are than being 5:100 DOX-PCL-PEG-COOH preparation
The preparation method of the DOX-PCL-PEG-COOH micellas as control experiment is present embodiments provided, it is specific as follows:
Take DOX powder 10mg molten in methyl alcohol, ultrasonic dissolution, be settled in 10mL volumetric flasks, obtain 1mg/mL DOX marks Quasi- product solution.Take 5mg PCL-PEG-COOH to be dissolved in 0.5mL acetone, obtain polymer solution 10mg/mL.100 μ L are taken to gather Polymer solution and 50 μ LDOX solution, add acetone until 500 μ L.Under conditions of being stirred vigorously, it is slowly added in 5mL pure water, stirs 4h is mixed, makes organic solvent evaporation.
The medicine of embodiment 4. and carrier dispensing are than being 10:The preparation of 100 DOX-PCL-PEG-COOH micellas
The preparation method of the DOX-PCL-PEG-COOH micellas as control experiment is present embodiments provided, it is specific as follows:
Take DOX powder 10mg molten in methyl alcohol, ultrasonic dissolution, be settled in 10mL volumetric flasks, obtain 1mg/mL DOX marks Quasi- product solution.Take 5mg PCL-PEG-COOH to be dissolved in 0.5mL acetone, obtain polymer solution 10mg/mL.Micellar drug and Micellar drug and carrier dispensing are than being 10:100 preparation:Take 100 μ L polymer solutions and 100 μ LDOX solution, add acetone until 500μL.Under conditions of being stirred vigorously, it is slowly added in 5mL pure water, stirs 4h, make organic solvent evaporation.
The medicine of embodiment 5. and carrier dispensing are than being 15:The preparation of 100 DOX-PCL-PEG-COOH micellas
The preparation method of the DOX-PCL-PEG-COOH micellas as control experiment is present embodiments provided, it is specific as follows:
Take DOX powder 10mg molten in methyl alcohol, ultrasonic dissolution, be settled in 10mL volumetric flasks, obtain 1mg/mL DOX marks Quasi- product solution.Take 5mg PCL-PEG-COOH to be dissolved in 0.5mL acetone, obtain polymer solution 10mg/mL.Micellar drug and Carrier dispensing is than being 15:100 preparation:100 μ L polymer solutions and 150 μ LDOX solution are taken, add acetone until 500 μ L.In play Under conditions of strong stirring, it is slowly added in 5mL pure water, stirs 4h, make organic solvent evaporation.
The preparation of the collagenase blank micellas of embodiment 6.PCL-PEG- IV
The preparation method of the collagenase blank micellas of PCL-PEG- IV as control experiment is present embodiments provided, It is specific as follows:
Precision weighs NHS (15.3mg, 0.15mmol) and EDC (28.7mg, 0.15mmol) is dissolved separately in 1mL dichloromethanes Alkane, which is vortexed, to be dissolved.Take COOH-PEG-PCL (50mg, 0.05mmol) solution to add 5mL dichloromethane, be placed in magnetic stirring apparatus In, after 20-26 DEG C of stirring 15min, while NHS and EDC solution are added, produce NHS-PEG-PCL, mix 6-8h, revolving So that the complete alkane volatilization of dichloromethane, adds pure water, produces product.Collagen solution is added in the NHS-PEG-PCL aqueous solution (1mL, 0.5mg/mL), stir 2h.Film dialysis 24h (Mw=3500), change water once per 6h, freeze 48h, obtain PCL-PEG- IV collagenase。
Take the collagenase of 5mg PCL-PEG- IV to be dissolved in 0.5mL acetone, obtain polymer solution 10mg/mL.Will 100 μ L polymer solutions and 400 μ L acetone are sufficiently mixed.Under conditions of being stirred vigorously, it is slowly added in 5mL pure water, stirs 4h, make organic solvent evaporation.
The medicine of embodiment 7. and carrier dispensing are than being 0.5:It is prepared by the 100 collagenase micellas of DOX-PCL-PEG- IV
The present invention's has the collagenase micellas of functionalization DOX-PCL-PEG- IV for penetrating tumor extracellular matrix effect Preparation method, it is specific as follows, wherein, in following examples, carrier refers both to be modified with the amphipathic nature polyalcohol of Collagenase:
Precision weighs NHS (15.3mg, 0.15mmol) and EDC (28.7mg, 0.15mmol) is dissolved separately in 1mL dichloromethanes Alkane, which is vortexed, to be dissolved.Take PCL-PEG-COOH (50mg, 0.05mmol) solution to add 5mL dichloromethane, be placed in magnetic stirring apparatus In, after 20-26 DEG C of stirring 15min, while NHS and EDC solution are added, produce NHS-PEG-PCL, mix 6-8h, revolving So that the complete alkane volatilization of dichloromethane, adds pure water, produces product.Type Ⅳ collagen albumen is added in the NHS-PEG-PCL aqueous solution Enzyme solutions (1mL, 0.5mg/mL), stir 2h.Film dialysis 24h (Mw=3500), change water once per 6h, freeze 48h, modified There is the amphipathic nature polyalcohol (collagenase of PCL-PEG- IV) of Collagenase.
Take DOX powder 10mg molten in methyl alcohol, ultrasonic dissolution, be settled in 100mL volumetric flasks, obtain 100 μ g/mL's DOX standard solutions.Take the collagenase of 5mg PCL-PEG- IV to be dissolved in 0.5mL acetone, obtain polymer solution 10mg/mL.100 μ L polymer solutions and 50 μ LDOX solution are taken, add acetone until 500 μ L.Under conditions of being stirred vigorously, delay It is slow to add in 5mL pure water, 4h is stirred, makes the amphipathic nature polyalcohol glue for obtaining penetrating tumor extracellular matrix after organic solvent evaporation Beam (the collagenase micellas of DOX-PCL-PEG- IV).
The medicine of embodiment 8. and carrier dispensing are than being 5:It is prepared by the 100 collagenase micellas of DOX-PCL-PEG- IV
Precision weighs NHS (15.3mg, 0.15mmol) and EDC (28.7mg, 0.15mmol) is dissolved separately in 1mL dichloromethanes Alkane, which is vortexed, to be dissolved.Take PCL-PEG-COOH (50mg, 0.05mmol) solution to add 5mL dichloromethane, be placed in magnetic stirring apparatus In, after 20-26 DEG C of stirring 15min, while NHS and EDC solution are added, produce NHS-PEG-PCL, mix 6-8h, revolving So that the complete alkane volatilization of dichloromethane, adds pure water, produces product.Collagen solution is added in the NHS-PEG-PCL aqueous solution (1mL, 0.5mg/mL), stir 2h.Film dialysis 24h (Mw=3500), change water once per 6h, freeze 48h, obtain PCL-PEG- IV collagenase。
Take DOX powder 10mg molten in methyl alcohol, ultrasonic dissolution, be settled in 10mL volumetric flasks, obtain 1mg/mL DOX marks Quasi- product solution.Take the collagenase of 5mg PCL-PEG- IV to be dissolved in 0.5mL acetone, obtain polymer solution 10mg/mL. 100 μ L polymer solutions and 50 μ L DOX solution are taken, add acetone until 500 μ L.Under conditions of being stirred vigorously, it is slowly added to In 5mL pure water, 4h is stirred, makes organic solvent evaporation.
The medicine of embodiment 9. and carrier dispensing are than being 10:It is prepared by the 100 collagenase micellas of PCL-PEG- IV
Precision weighs NHS (15.3mg, 0.15mmol) and EDC (28.7mg, 0.15mmol) is dissolved separately in 1mL dichloromethanes Alkane, which is vortexed, to be dissolved.Take PCL-PEG-COOH (50mg, 0.05mmol) solution to add 5mL dichloromethane, be placed in magnetic stirring apparatus In, after 20-26 DEG C of stirring 15min, while NHS and EDC solution are added, produce NHS-PEG-PCL, mix 6-8h, revolving So that the complete alkane volatilization of dichloromethane, adds pure water, produces product.Collagen solution is added in the NHS-PEG-PCL aqueous solution (1mL, 0.5mg/mL), stir 2h.Film dialysis 24h (Mw=3500), change water once per 6h, freeze 48h, obtain PCL-PEG- IV collagenase。
Take DOX powder 10mg molten in methyl alcohol, ultrasonic dissolution, be settled in 10mL volumetric flasks, obtain 1mg/mL DOX marks Quasi- product solution.Take the collagenase of 5mg PCL-PEG- IV to be dissolved in 0.5mL acetone, obtain polymer solution 10mg/mL. 100 μ L polymer solutions and 100 μ LDOX solution are taken, add acetone until 500 μ L.Under conditions of being stirred vigorously, it is slowly added to In 5mL pure water, 4h is stirred, makes organic solvent evaporation.
The medicine of embodiment 10. and carrier dispensing are than being 15:It is prepared by the 100 collagenase micellas of PCL-PEG- IV
Precision weighs NHS (15.3mg, 0.15mmol) and EDC (28.7mg, 0.15mmol) is dissolved separately in 1mL dichloromethanes Alkane, which is vortexed, to be dissolved.Take PCL-PEG-COOH (50mg, 0.05mmol) solution to add 5mL dichloromethane, be placed in magnetic stirring apparatus In, after 20-26 DEG C of stirring 15min, while NHS and EDC solution are added, produce NHS-PEG-PCL, mix 6-8h, revolving So that the complete alkane volatilization of dichloromethane, adds pure water, produces product.Collagen solution is added in the NHS-PEG-PCL aqueous solution (1mL, 0.5mg/mL), stir 2h.Film dialysis 24h (Mw=3500), change water once per 6h, freeze 48h, obtain PCL-PEG- IV collagenase。
Take DOX powder 10mg molten in methyl alcohol, ultrasonic dissolution, be settled in 10mL volumetric flasks, obtain 1mg/mL DOX marks Quasi- product solution.Take the collagenase of 5mg PCL-PEG- IV to be dissolved in 0.5mL acetone, obtain polymer solution 10mg/mL. 100 μ L polymer solutions and 150 μ LDOX solution are taken, add acetone until 500 μ L.Under conditions of being stirred vigorously, it is slowly added to In 5mL pure water, 4h is stirred, makes organic solvent evaporation.
The particle diameter of embodiment 11. and potential measurement
Micellar solution in embodiment 1,2,6 and 7 is diluted to certain concentration with pure water, by particle size analyzer and The particle diameter distribution and surface potential of Zeta potential analysis-e/or determining micella.
Table 1 is the particle diameter distribution of embodiment 1,2,6 and 7 and the result of surface potential, and PCL-PEG- is can be seen that from form The particle diameter of COOH blank micellas increases to 180nm or so after 110nm or so, load medicine.PCL-PEG-Ⅳcollagenase The particle diameter of micella increases to 360nm or so after 310nm or so, load medicine.Simultaneously, it can be seen that either before load medicine still After carrying medicine, current potential is in 0mV or so, in electroneutral.
The particle diameter distribution and Surface potential measurement result of 1 different micellar solutions of table
The morphologic observation of the micella of embodiment 12.
Take the micella of the embodiment 1,2,6 and 7 prepared to drop to respectively to be covered with the copper mesh for supporting film, be placed under infrared lamp 2-4h is dried, the form of nanoparticle is observed using transmission electron microscope method and taken pictures.
Fig. 1 is the test result of above-described embodiment, and figure a, b, c, d represent the micella of embodiment 1,2,6 and 7 respectively.Pass through Micella in figure a and b and the micella in figure c and d are contrasted, increased it may be seen that carrying particle diameter after medicine, but form remains unchanged It is regular circle, drug adriamycin enters carrier inside.
The micella envelop rate of embodiment 13. and drugloading rate measure
Micellar solution in Example 3-5 and 8-10, envelop rate and drugloading rate are determined using supercentrifugal process, will be made The micella got ready, is placed in centrifuge tube, under the conditions of 8000rap/min, 4 DEG C, ultracentrifugation 30min, takes supernatant, measure trip From medicament contg.
10mg DOX is weighed, adds pure water, 100mL is settled to after ultrasonic dissolution, obtains 100 μ g/mL DOX aqueous solution marks Quasi- product are made into a series of gradient concentrations, prepare standard curve, and the fluorescent value of determination sample.
The envelop rate and drugloading rate of micella are calculated by following equation:
Drugloading rate (%)=WDOX/WCarrier× 100%
Envelop rate (%)=(WDOX-WIt is free)/WTotal DOX× 100%
Wherein, WTotal DOXIt is medicine total amount, WIt is freeIt is not wrap the quality into the medicine of micella, WCarrierIt is the quality of carrier.
Table 2 is embodiment 3-5 and 8-10 envelop rate and drugloading rate result, when the rate of feeding intake is 15%, two kinds of carriers There is muddiness in solution, so select to feed intake rate for 10% when dialysis measure envelop rate and drugloading rate.As can be seen from Table 2, DOX-PCL-PEG-COOH carriers envelop rate is 90%, and drugloading rate is 9%, after connecting IV collagenase, envelop rate and carries medicine Amount slightly reduces.
The envelop rate and drugloading rate test result of 2 different micellas of table
The vitro release of embodiment 14. is investigated
Bag filter boils 15min with 60 DEG C of pure water, and the DOX-PCL-PEG-COOH micellar solutions of embodiment 4 are placed in into bag filter In, tightened, be suspended in the 100mL beakers for the PBS for filling 40mL pH7.4 with cotton cord, is put in constant temperature oscillation case with 37 ± 1 DEG C, 48h, timing 0.5,1,2,4,8,12,24,48 and 72h sampling 1mL are vibrated, while add isometric synthermal drug release and be situated between Matter PBS.By samples taken, drug release amount is determined, calculates cumulative release percentage.
Fig. 2 is the test result of the micella prepared by embodiment 4, it is seen that with the increase of release time, medicine The Accumulation dissolution of thing gradually rises, and after the time is more than 72h, release profiles tend to be steady, it can be seen that DOX-PCL-PEG- COOH has obvious slow releasing function.
The medicine of embodiment 15. and carrier dispensing are than being 0.5:It is prepared by 100 DOX-PLA-PEG-COOH micellas
Amphipathic nature polyalcohol in embodiment 7 is replaced with into PLA-PEG-COOH, other experimental procedures referring to embodiment 7, The amphipathic nature polyalcohol micella of tumor extracellular matrix can be penetrated by being made.
The medicine of embodiment 16. and carrier dispensing are than being 0.5:It is prepared by 100 DOX-PLGA-PEG-COOH micellas
Amphipathic nature polyalcohol in embodiment 7 is replaced with into PLGA-PEG-COOH, other experimental procedures referring to embodiment 7, The amphipathic nature polyalcohol micella of tumor extracellular matrix can be penetrated by being made.
The medicine of embodiment 17. and carrier dispensing are than being 5:It is prepared by the 100 collagenase micellas of DTX-PCL-PEG- I
Antineoplastic in embodiment 8 is replaced with into-DTX, IV collagenase replaces with I collagenase, its The amphipathic nature polyalcohol micella that can penetrate tumor extracellular matrix is made referring to embodiment 8 in his experimental procedure.
The medicine of embodiment 18. and carrier dispensing are than being 5:It is prepared by the 100 collagenase micellas of TAXOL-PCL-PEG- II
Antineoplastic in embodiment 8 is replaced with into TAXOL, IV collagenase replaces with II collagenase, The amphipathic nature polyalcohol micella that can penetrate tumor extracellular matrix is made referring to embodiment 8 in other experimental procedures.
The medicine of embodiment 19. and carrier dispensing are than being 5:It is prepared by the 100 collagenase micellas of TAXOL-PCL-PEG- V
Antineoplastic in embodiment 8 is replaced with into TAXOL, IV collagenase replaces with V collagenase, The amphipathic nature polyalcohol micella that can penetrate tumor extracellular matrix is made referring to embodiment 8 in other experimental procedures.
Described above is only the preferred embodiment of the present invention, is not intended to limit the invention, it is noted that for this skill For the those of ordinary skill in art field, without departing from the technical principles of the invention, can also make it is some improvement and Modification, these improvement and modification also should be regarded as protection scope of the present invention.

Claims (10)

  1. A kind of 1. amphipathic nature polyalcohol micella for penetrating tumor extracellular matrix, it is characterised in that:Including antineoplastic, amphiphilic The antineoplastic is wrapped in inside it by property polymer and Collagenase, the amphipathic nature polyalcohol, the amphiphilic Property polymer outer passes through Collagenase described in covalent key connection.
  2. 2. the amphipathic nature polyalcohol micella according to claim 1 for penetrating tumor extracellular matrix, it is characterised in that:It is described Antineoplastic is the one or more in docetaxel, adriamycin and taxol.
  3. 3. the amphipathic nature polyalcohol micella according to claim 1 for penetrating tumor extracellular matrix, it is characterised in that:It is described Amphipathic nature polyalcohol is the one or more in PCL-PEG copolymers, PLA-PEG copolymers and PLGA-PEG copolymers.
  4. 4. the amphipathic nature polyalcohol micella according to claim 1 for penetrating tumor extracellular matrix, it is characterised in that:It is described The mol ratio of Collagenase and amphipathic nature polyalcohol is 1:100-1000.
  5. 5. the amphipathic nature polyalcohol micella according to claim 1 for penetrating tumor extracellular matrix, it is characterised in that:It is described Collagenase is one kind in NTx protease, II Collagenase, type Ⅳ collagen protease and V collagen type enzyme It is or several.
  6. 6. a kind of amphipathic nature polyalcohol micella for penetrating tumor extracellular matrix according to any one of claim 1-5 Preparation method, it is characterised in that comprise the following steps:
    (1) amphipathic nature polyalcohol and crosslinking agent are reacted at 20-26 DEG C in the first organic solvent, removed described First organic solvent, then reacted in pure water with Collagenase at 20-26 DEG C, obtain being modified with the two of Collagenase Parent's property polymer;
    (2) amphipathic nature polyalcohol for being modified with Collagenase and the antineoplastic are dissolved in the second organic solvent, Obtain drug solution;
    (3) drug solution is added to the water, removes second organic solvent, tumor extracellular matrix can be penetrated described in formation Amphipathic nature polyalcohol micella.
  7. 7. preparation method according to claim 6, it is characterised in that:In step (1), the crosslinking agent is N- hydroxyl ambers Amber acid imide and 1- ethyls-(3- dimethylaminopropyls) carbodiimide hydrochloride.
  8. 8. preparation method according to claim 6, it is characterised in that:In step (1), first organic solvent is two The one or more of first sulfoxide, acetonitrile and chloroform.
  9. 9. preparation method according to claim 6, it is characterised in that:In step (2), second organic solvent is third Ketone, methanol, ethanol, the one or more of dichloromethane and chloroform.
  10. 10. preparation method according to claim 6, it is characterised in that:In step (3), under agitation by described in Drug solution is added in pure water, mixing speed 400-700rpm.
CN201711020966.1A 2017-10-26 2017-10-26 Amphipathic nature polyalcohol micella of tumor extracellular matrix and preparation method thereof can be penetrated Pending CN107714646A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201711020966.1A CN107714646A (en) 2017-10-26 2017-10-26 Amphipathic nature polyalcohol micella of tumor extracellular matrix and preparation method thereof can be penetrated

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201711020966.1A CN107714646A (en) 2017-10-26 2017-10-26 Amphipathic nature polyalcohol micella of tumor extracellular matrix and preparation method thereof can be penetrated

Publications (1)

Publication Number Publication Date
CN107714646A true CN107714646A (en) 2018-02-23

Family

ID=61201759

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201711020966.1A Pending CN107714646A (en) 2017-10-26 2017-10-26 Amphipathic nature polyalcohol micella of tumor extracellular matrix and preparation method thereof can be penetrated

Country Status (1)

Country Link
CN (1) CN107714646A (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113425682A (en) * 2021-08-03 2021-09-24 宁夏医科大学 Drug targeting polymeric micelle and preparation method and application thereof
WO2024109964A1 (en) * 2022-11-23 2024-05-30 上海交通大学医学院附属第九人民医院 Oral drug-loaded micelle composition and preparation method therefor

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101787119A (en) * 2010-03-25 2010-07-28 复旦大学 Polymer with tumor organization pH responsiveness and micelle thereof
CN103977434A (en) * 2013-02-07 2014-08-13 复旦大学 P-hydroxybenzoic acid mediated polymer micelle drug delivering system with brain targeting function
CN104274834A (en) * 2013-07-08 2015-01-14 复旦大学 Environment-sensitive tumor-targeting polymer micelle and preparation method thereof
CN106265509A (en) * 2016-08-10 2017-01-04 国家纳米科学中心 A kind of pH and Redox double-bang firecracker answers amphipathic nature block polymer and its production and use
CN106317416A (en) * 2016-09-07 2017-01-11 国家纳米科学中心 Double-pH-response amphiphilic copolymer and preparation method and application thereof
CN106581686A (en) * 2016-12-15 2017-04-26 中国药科大学 Preparation and application of hyaluronic acid-modified amphipathic chitosan derivative carrier with tumor microenvironment specificity drug release effect
CN106727313A (en) * 2017-01-06 2017-05-31 国家纳米科学中心 A kind of drug-carrying polymer nano-micelle and its preparation method and application
CN106963756A (en) * 2017-04-05 2017-07-21 中国药科大学 Pharmaceutical polymer micella and its application in pharmacy are carried altogether

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101787119A (en) * 2010-03-25 2010-07-28 复旦大学 Polymer with tumor organization pH responsiveness and micelle thereof
CN103977434A (en) * 2013-02-07 2014-08-13 复旦大学 P-hydroxybenzoic acid mediated polymer micelle drug delivering system with brain targeting function
CN104274834A (en) * 2013-07-08 2015-01-14 复旦大学 Environment-sensitive tumor-targeting polymer micelle and preparation method thereof
CN106265509A (en) * 2016-08-10 2017-01-04 国家纳米科学中心 A kind of pH and Redox double-bang firecracker answers amphipathic nature block polymer and its production and use
CN106317416A (en) * 2016-09-07 2017-01-11 国家纳米科学中心 Double-pH-response amphiphilic copolymer and preparation method and application thereof
CN106581686A (en) * 2016-12-15 2017-04-26 中国药科大学 Preparation and application of hyaluronic acid-modified amphipathic chitosan derivative carrier with tumor microenvironment specificity drug release effect
CN106727313A (en) * 2017-01-06 2017-05-31 国家纳米科学中心 A kind of drug-carrying polymer nano-micelle and its preparation method and application
CN106963756A (en) * 2017-04-05 2017-07-21 中国药科大学 Pharmaceutical polymer micella and its application in pharmacy are carried altogether

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
NGUYEN-VAN CUONG ETAL: ""Doxorubicin-Loaded PEG-PCL-PEG Micelle Using Xenograft Model of Nude Mice: Effect of Multiple Administration of Micelle on the Suppression of Human Breast Cancer"", 《CANCERS》 *
SURYA MURTY ETAL: ""Nanoparticles functionalized with collagenase exhibit improved tumor accumulation in a murine xenograft model"", 《PART PART SYST CHARACT》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113425682A (en) * 2021-08-03 2021-09-24 宁夏医科大学 Drug targeting polymeric micelle and preparation method and application thereof
WO2024109964A1 (en) * 2022-11-23 2024-05-30 上海交通大学医学院附属第九人民医院 Oral drug-loaded micelle composition and preparation method therefor

Similar Documents

Publication Publication Date Title
Sun et al. Bioreducible PAA-g-PEG graft micelles with high doxorubicin loading for targeted antitumor effect against mouse breast carcinoma
Li et al. Redox-sensitive micelles self-assembled from amphiphilic hyaluronic acid-deoxycholic acid conjugates for targeted intracellular delivery of paclitaxel
CN104177624B (en) Dual Sensitive amphipathic three block copolymer containing disulfide bond and acylhydrazone key and preparation method and application
CN107095859B (en) Drug-loaded nanocapsule with tumor cell bioreductive microenvironment sensitivity and preparation method thereof
JP2014518862A (en) Polymer nanoparticles for drug delivery
KR100837860B1 (en) Hydrophilic polymer nanocapsules and method for preparing the same
CN105853403B (en) A kind of paclitaxel palmitate liposome and preparation method thereof
WO2012040513A1 (en) Compositions and methods for the delivery of beta lapachone
Shang et al. Enhancing cancer chemo-immunotherapy by biomimetic nanogel with tumor targeting capacity and rapid drug-releasing in tumor microenvironment
Ji et al. Curcumin‐loaded mixed micelles: Preparation, characterization, and in vitro antitumor activity
Fathi Karkan et al. Cisplatin-loaded superparamagnetic nanoparticles modified with PCL-PEG copolymers as a treatment of A549 lung cancer cells
Shuai et al. Self-assembling poly (ethylene glycol)-block-polylactide-cabazitaxel conjugate nanoparticles for anticancer therapy with high efficacy and low in vivo toxicity
EP2978423A1 (en) Stable nanocomposition comprising doxorubicin, process for the preparation thereof, its use and pharmaceutical compositions containing it
CN104888235A (en) pH sensitive nanoparticles prodrug with capacity of co-delivering multiple drugs, preparation method and application thereof
EP2978420A1 (en) Stable nanocomposition comprising paclitaxel, process for the preparation thereof, its use and pharmaceutical compositions containing it
KR20180097707A (en) Biodegradable amphipathic polymers, polymeric vehicles prepared thereby, and use in the manufacture of a therapeutic agent for lung cancer target
CN104490786B (en) Preparation method and application of targeted multi-function double drug-loading liposome
CN113633625B (en) Nano-drug of hybrid membrane loaded oxidative phosphorylation inhibitor and preparation method thereof
US20140296173A1 (en) Stable nanocomposition comprising epirubicin, process for the preparation thereof, its use and pharmaceutical compositions containing it
Ghazal et al. Role of nanoparticles in enhancing chemotherapy efficacy for cancer treatment
US20220305043A1 (en) Formulations of cyclic macromolecule-based nanoparticles encapsulating small molecules
CN107714646A (en) Amphipathic nature polyalcohol micella of tumor extracellular matrix and preparation method thereof can be penetrated
CN105233282B (en) A kind of multifunctional nano pharmaceutical composition and preparation method thereof
Xu et al. Development, Characterization, and Evaluation of PSMA‐Targeted Glycol Chitosan Micelles for Prostate Cancer Therapy
CN107126425A (en) A kind of tanshinone IIA PEG PLGA PEG nanoparticles and preparation method thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20180223