CN107308461A - A kind of nano-particle of colon tumor cell active targeting and preparation method thereof - Google Patents
A kind of nano-particle of colon tumor cell active targeting and preparation method thereof Download PDFInfo
- Publication number
- CN107308461A CN107308461A CN201710563301.9A CN201710563301A CN107308461A CN 107308461 A CN107308461 A CN 107308461A CN 201710563301 A CN201710563301 A CN 201710563301A CN 107308461 A CN107308461 A CN 107308461A
- Authority
- CN
- China
- Prior art keywords
- particle
- nano
- preparation
- solution
- polyvinyl alcohol
- 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
Links
Classifications
-
- 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
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a kind of nano-particle of colon tumor cell active targeting and preparation method thereof.The nano-particle for carrying camptothecine is prepared by oil-in-water solvent evaporation method first, targeting modification then is carried out with chondroitin sulfate to this nanoparticle surface.The particle diameter of the nano-particle is 289.5 ± 5.51 nm, is observed by SEM, and nano-particle is rounded or ellipse, and size is more uniform.Chondroitin sulfate is introduced nanoparticle surface by the present invention, and this particle with the CD44 cytokines that cancer cell surfaces are excessively expressed by being combined, and this analgesics screening platform mode adds the phagocytosis efficiency of colon cancer cell.Experimental result is shown：The nano-particle of chondroitin sulfate modification significantly improves the suppression efficiency of colon cancer cell phagocytosis efficiency and cell growth, so as to effectively enhance the therapeutic efficiency of colon cancer.Preparation technology of the present invention is simply, efficiently, stably, simple and efficient to handle.
Description
Technical field
The invention belongs to nano-medicament carrier technical field, and in particular to a kind of nanoparticle targetted available for tumour cell
Sub- preparation method, sign and application.
Background technology
Tumour is a kind of common disease, frequently-occurring disease, seriously threatens the life security of the mankind.Colon cancer has become main
Public health problem, more than 1,400,000 new cases, accounts for more than half of the annual millions of people's death in the whole world.According to Britain in 2012
There are 14,100,000 new cancer cases in Cancer center's data display, the whole world, wherein there is 8,200,000 cancer deaths.At present, clinically
Treatment tumour common method has chemotherapy, radiotherapy and surgery excision etc..Wherein, chemotherapy is clinically conventional cancer
Disease treatment method, but classic chemotherapy mode has strong side effect to health tissues, cell, such as bone marrow suppression, catarrh and de-
Hair etc..At present, chemotherapeutic significant challenge is development drug delivery system, for safely, effectively delivering chemotherapeutics.
The drug delivery system of nanostructured is mainly by intravenously administrable, including nano particle, liposome and micella etc..It is logical
The approach is crossed, there are many serious Problems Existings sufficiently to solve, such as inorganizable specificity and tumor locus are inefficient
Insoluble drug release, these all make chemotherapy effect not satisfactory.
In recent years, with Poly(D,L-lactide-co-glycolide（Polylactic-co-glycolic acid, PLGA）For base
Micro-, the nano-particle of plinth have obtained extensive research, its have good biocompatibility, nonirritant, non-immunogenicity and
The advantages of medicine controlled releasing.Camptothecine (CPT) is a kind of alkaloid extracted from natural plants, can suppress ribozyme topoisomerase
Enzyme I, is a kind of antitumor agent got a good chance of.But its high hydrophobicity and stability is poor, these defects cause it not yet big
Sizable application is in clinic.
Therefore, in order to provide a practicable therapeutic scheme to oncotherapy, we are prepared into PLGA parcels CPT
Nano-particle, while in particle surface modified chondroitin sulfate, compared to more traditional without targeted nano-particle, chondroitin sulfate is repaiied
Faster, antitumous effect is more preferable, as a kind of potential ideas of cancer therapy for the nano-particle drug cell phagocytosis of decorations.
The content of the invention
In view of this, an object of the present invention is to provide a kind of nano-particle targetted available for tumour cell, its
With good biocompatibility, stability, available for the treatment of colon cancer, the second object of the present invention is to provide above-mentioned target
To the preparation method of nano-particle.
For achieving the above object, technical scheme is：
The preparation method for carrying medicine targeted nano-particle comprises the following steps：
（1）Weigh Poly(D,L-lactide-co-glycolide（Polylactic-co-glycolic acid, PLGA）And camptothecine
（CPT）, it is dissolved in dichloromethane and methanol（Methylene chloride/methanol=4:1）, wiring solution-forming；
（2）By step（1）Obtain solution to be added dropwise in polyvinyl alcohol water solution, be made into emulsion；
（3）By step（2）Middle gained emulsion is immediately placed in ice bath, carries out ultrasound；
（4）Emulsion after ultrasound is poured into polyvinyl alcohol water solution rapidly；
（5）At ambient temperature, the organic solvent in emulsion step (4) obtained fully volatilizees；
（6）Centrifuged again, collect nano-particle, and be washed with deionized 3 times；
（7）By step（6）Obtained nano-particle is suspended in the sodium-acetate buffer of pH=6, adds the sulphur containing EDC and NHS
Aching and limp ossein solution, can obtain the nano-particle of surface modification chondroitin sulfate；
（8）The nano-particle of surface modification chondroitin sulfate is collected by centrifugation again, and is washed with deionized 3 times；
（9）By step（8）Obtained nano-particle is dispersed in the aqueous solution of trehalose again；
（10）Above-mentioned nano-particle is freezed again, -20 degrees Celsius of refrigerators are stored in standby.
It is preferred that, the step（1）In solution in PLGA mass concentration for 25-150 mg/mL, CPT quality it is dense
Spend for 2-10mg/mL.
It is preferred that, the step（2）The mass concentration of polyvinyl alcohol is 1-5%, polyvinyl alcohol in middle polyvinyl alcohol water solution
The volume of the aqueous solution is step（1）2 times of obtained solution..
It is preferred that, the step（3）In, ultrasonic parameter is ultrasonic amplitude 20 ~ 50%, every time 5 ~ 20 s, is spaced 1 ~ 5 s,
Ultrasonic total duration is 0.5 ~ 3 min.
It is preferred that, the step（4）In, the mass concentration of polyvinyl alcohol is 0 ~ 0.5%, volume in polyvinyl alcohol water solution
For step（3）10-60 times of obtained emulsion volume.
It is preferred that, the step（6）And step（8）In, the parameter that centrifugation is chosen is that centrifugation rate is 12000 rpm, from
The heart time is 15-20 min.
It is preferred that, the step（9）In, trehalose concentration is 10-20% in aqueous trehalose, and nano-particle is dissolved in marine alga
PLGA mass concentration is 25-150 mg/mL after sugar aqueous solution.
It is preferred that, the step（10）In, freeze-drying is carried out in accordance with the following methods：After -80 degrees Celsius of freeze overnights
It is placed in freeze dryer and freezes 12~36 h.
Relative to existing technology, the advantage of the invention is that：
1. the present invention is using PLGA as drug carrier material, preparation method is oil-in-water solvent evaporation method, whole preparation process letter
Single quick, short preparation period, yield is high.
2. the present invention wraps up CPT by PLGA, medicine-carried nano particles are prepared, improve CPT water-soluble not good, unstable
Deng, CPT, which is preferably delivered to patient part, plays a role, increase curative effect.
3. invention introduces chondroitin sulfate as targeted molecular, the nano-particle of chondroitin sulfate modification by with
The CD44 cytokines that cancer cell surfaces are excessively expressed are combined, and this analgesics screening platform mode significantly improves colon
The phagocytosis efficiency and antitumous effect of cancer cell, so as to effectively enhance the therapeutic efficiency of colon cancer.
Brief description of the drawings
In order that the object, technical solutions and advantages of the present invention are clearer, below in conjunction with accompanying drawing the present invention is made into
The detailed description of one step, wherein：
Fig. 1：The field emission scanning electron microscope of the nano-particle of carboxymethyl cellulose modification and the nano-particle of chondroitin sulfate modification
Figure.
Fig. 2：The external phagocytosis of the nano-particle of carboxymethyl cellulose modification and the nano-particle of chondroitin sulfate modification is real
Test.
Fig. 3：The extracorporeal anti-tumor of the nano-particle of carboxymethyl cellulose modification and the nano-particle of chondroitin sulfate modification
Experiment.
Embodiment
Hereinafter with reference to accompanying drawing, the preferred embodiments of the present invention are described in detail.
Embodiment 1
Embodiment 1 is the preparation method for the load CPT nano-particles that chondroitin sulfate is modified, and is comprised the following steps：
（1）The mg of the PLGA 100 and mg of CPT 6 are weighed to be dissolved in jointly in 2 mL dichloromethane and methanol solution；
（2）Above-mentioned organic solution is added dropwise in the poly-vinyl alcohol solutions of 4 mL 5%；
（3）Above-mentioned solution is immediately placed in ice bath, carries out ultrasonic, amplitude 50%, every time 10 s, is spaced 1s, and ultrasonic total duration is 1
min；
（4）After ultrasound is finished, in the poly-vinyl alcohol solution that above-mentioned emulsion is poured into rapidly to 60 mL 0.05%；
（5）At ambient temperature, organic solvent in above-mentioned emulsion is volatilized；
（7）Above-mentioned nano-particle is collected by centrifugation, and is washed with deionized 3 times, centrifugation rate is 12000 rpm, centrifugation time
For 20 min；
（8）Made particle and chondroitin sulfate solution are reacted at room temperature 2 hours；
（9）Above-mentioned nano-particle is collected by centrifugation, and is washed with deionized 3 times, centrifugation rate is 12000 rpm, centrifugation time
For 20 min；
（10）Above-mentioned nano-particle is dispersed in the aqueous solution of 1 mL containing 10% trehalose again；
（11）Above-mentioned nano-particle is freezed, -20 DEG C of refrigerators are stored in；
Embodiment 2
Embodiment 2 is the preparation method for the load CPT nano-particles that carboxymethyl cellulose is modified, and is comprised the following steps：
（1）The mg of the PLGA 100 and mg of CPT 6 are weighed to be dissolved in jointly in 2 mL dichloromethane and methanol solution；
（2）Above-mentioned organic solution is added dropwise in the poly-vinyl alcohol solutions of 4 mL 5%；
（3）Above-mentioned solution is immediately placed in ice bath, carries out ultrasonic, amplitude 50%, every time 10 s, is spaced 1s, and ultrasonic total duration is 1
min；
（4）After ultrasound is finished, in the poly-vinyl alcohol solution that above-mentioned emulsion is poured into rapidly to 60 mL 0.05%；
（5）At ambient temperature, organic solvent in above-mentioned emulsion is volatilized；
（7）Above-mentioned nano-particle is collected by centrifugation, and is washed with deionized 3 times, centrifugation rate is 12000 rpm, centrifugation time
For 20 min；
（8）Made particle and cmc soln are reacted at room temperature 2 hours；
（9）Above-mentioned nano-particle is collected by centrifugation, and is washed with deionized 3 times, centrifugation rate is 12000 rpm, centrifugation time
For 20 min；
（10）Above-mentioned nano-particle is dispersed in the aqueous solution of 1 mL containing 10% trehalose again；
（11）Above-mentioned nano-particle is freezed, -20 DEG C of refrigerators are stored in；
Experiment 1：The pattern of nano-particle is observed by field emission scanning electron microscope.Before analysis, nano-particle passes through
Vacuum spray platinum processing.
Fig. 1 is the field emission scanning electron microscope figure of nano-particle.（a）Figure is the load CPT particles of carboxymethyl cellulose modification；
（b）Figure is the load CPT particles of chondroitin sulfate modification.Obtained nano-particle is can be seen that in ball from obtained electromicroscopic photograph
Shape, surface is smooth, and pore size is more uniform.
Experiment 2：Cell phagocytosis experiment, cell used is CT26WT, as can be seen from Figure 2 with receiving that aching and limp ossein is modified
The phagocytosis of grain of rice daughter cell is more efficient.
Experiment 3：Cell toxicity test, cell used is CT26WT, as shown in figure 3, two kinds of nano-particles are total to cell
Cultivate after 24h, 48h, plus MTT is cultivated 4 hours, ELIASA determines OD values at 570nm.It will become apparent from using chondroitin sulfate from figure
The nano-particle of element modification has more preferable inhibition to tumour cell, and than 24 hours 48 hours inhibitions are obvious.
Finally illustrate, above example is only unrestricted for illustrating technical scheme, although passing through ginseng
According to the preferred embodiments of the present invention, invention has been described, it should be appreciated by those of ordinary skill in the art that can
So that various changes are made to it in the form and details, the present invention limited without departing from appended claims
Spirit and scope.
Claims (8)
1. a kind of nano-particle of colon tumor cell active targeting and preparation method thereof, it is characterised in that：Volatilized using solvent
Method prepares nano-particle, comprises the following steps：
（1）Weigh Poly(D,L-lactide-co-glycolide（Polylactic-co-glycolic acid, PLGA）And camptothecine
（CPT）, it is dissolved in dichloromethane and methanol（Methylene chloride/methanol=4:1）, wiring solution-forming；
（2）By step（1）Obtain solution to be added dropwise in polyvinyl alcohol water solution, be made into emulsion；
（3）By step（2）Middle gained emulsion is immediately placed in ice bath, carries out ultrasound；
（4）Emulsion after ultrasound is poured into polyvinyl alcohol water solution rapidly；
（5）At ambient temperature, by step（4）Organic solvent in obtained emulsion fully volatilizees；
（6）Centrifuged again, collect nano-particle, and be washed with deionized 3 times；
（7）By step（6）Obtained nano-particle is suspended in the sodium-acetate buffer of pH=6, adds the sulphur containing EDC and NHS
Aching and limp ossein solution, can obtain the nano-particle of surface modification chondroitin sulfate；
（8）The nano-particle of surface modification chondroitin sulfate is collected by centrifugation again, and is washed with deionized 3 times；
（9）By step（8）Obtained nano-particle is dispersed in the aqueous solution of trehalose again；
（10）Above-mentioned nano-particle is freezed again, -20 degrees Celsius of refrigerators are stored in standby.
2. it is according to claim 1 a kind of for nano-particle of macrophage targeted delivery of drugs and preparation method thereof,
It is characterized in that：The step（1）In solution in PLGA mass concentration be for 25-150 mg/mL, CPT mass concentration
2-10 mg/mL。
3. it is according to claim 1 a kind of for nano-particle of macrophage targeted delivery of drugs and preparation method thereof,
It is characterized in that：The step（2）The mass concentration of polyvinyl alcohol is 1-5% in middle polyvinyl alcohol water solution, and polyvinyl alcohol is water-soluble
The volume of liquid is step（1）2 times of obtained solution.
4. it is according to claim 1 a kind of for nano-particle of macrophage targeted delivery of drugs and preparation method thereof,
It is characterized in that：The step（3）In, ultrasonic parameter is ultrasonic amplitude 20 ~ 50%, every time 5 ~ 20 s, is spaced 1 ~ 5 s, ultrasound
Total duration is 0.5 ~ 3 min.
5. it is according to claim 1 a kind of for nano-particle of macrophage targeted delivery of drugs and preparation method thereof,
It is characterized in that：The step（4）In, the mass concentration of polyvinyl alcohol is 0 ~ 0.5% in polyvinyl alcohol water solution, and volume is step
Suddenly（3）10-60 times of obtained emulsion volume.
6. it is according to claim 1 a kind of for nano-particle of macrophage targeted delivery of drugs and preparation method thereof,
It is characterized in that：The step（6）And step（8）In, the parameter that centrifugation is chosen is that centrifugation rate is 12000 rpm, during centrifugation
Between be 15-20 min.
7. it is according to claim 1 a kind of for nano-particle of macrophage targeted delivery of drugs and preparation method thereof,
It is characterized in that：The step（9）In, trehalose concentration is 10-20% in aqueous trehalose, and nano-particle is dissolved in marine alga syrup
PLGA mass concentration is 25-150 mg/mL after solution.
8. it is according to claim 1 a kind of for nano-particle of macrophage targeted delivery of drugs and preparation method thereof,
It is characterized in that：The step（10）In, freeze-drying is carried out in accordance with the following methods：It is placed in after -80 degrees Celsius of freeze overnights
12~36 h are freezed in freeze dryer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710563301.9A CN107308461A (en) | 2017-07-12 | 2017-07-12 | A kind of nano-particle of colon tumor cell active targeting and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710563301.9A CN107308461A (en) | 2017-07-12 | 2017-07-12 | A kind of nano-particle of colon tumor cell active targeting and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107308461A true CN107308461A (en) | 2017-11-03 |
Family
ID=60177610
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710563301.9A Pending CN107308461A (en) | 2017-07-12 | 2017-07-12 | A kind of nano-particle of colon tumor cell active targeting and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107308461A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108743540A (en) * | 2018-06-20 | 2018-11-06 | 西南大学 | A kind of preparation method and applications of the multifunctional nano drug based on fibroin |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106727429A (en) * | 2016-12-30 | 2017-05-31 | 西南大学 | A kind of targeting anti-tumor Nano medication for suppressing multidrug-resisting gene expression |
| CN106727430A (en) * | 2017-01-04 | 2017-05-31 | 西南大学 | A kind of preparation method for carrying medicine porous nano particle |
| CN106822037A (en) * | 2016-12-30 | 2017-06-13 | 西南大学 | A kind of preparation method of the nano-particle of the tripeptides modification for targetting colitis tissue |
-
2017
- 2017-07-12 CN CN201710563301.9A patent/CN107308461A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106727429A (en) * | 2016-12-30 | 2017-05-31 | 西南大学 | A kind of targeting anti-tumor Nano medication for suppressing multidrug-resisting gene expression |
| CN106822037A (en) * | 2016-12-30 | 2017-06-13 | 西南大学 | A kind of preparation method of the nano-particle of the tripeptides modification for targetting colitis tissue |
| CN106727430A (en) * | 2017-01-04 | 2017-05-31 | 西南大学 | A kind of preparation method for carrying medicine porous nano particle |
Non-Patent Citations (2)
| Title |
|---|
| BO XIAO等: "Hyaluronic acid-functionalized polymeric nanoparticles for colon cancer-targeted combination chemotherapy", 《NANOSCALE》 * |
| MENGHANG ZU等: "Chondroitin sulfate-functionalized polymeric nanoparticles for colon cancer-targeted chemotherapy", 《COLLOIDS AND SURFACES B: BIOINTERFACES》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108743540A (en) * | 2018-06-20 | 2018-11-06 | 西南大学 | A kind of preparation method and applications of the multifunctional nano drug based on fibroin |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109078183B (en) | Bionic multifunctional nano preparation based on cancer cell membrane and preparation method and application thereof | |
| Chen et al. | Oral delivery of curcumin via porous polymeric nanoparticles for effective ulcerative colitis therapy | |
| CN109731106B (en) | Preparation method of compound for treating brain glioma | |
| CN104586775B (en) | Astragalus polysaccharide sustained release microsphere for treating radiation pneumonitis and preparation method of astragalus polysaccharide sustained release microsphere | |
| CN112716915A (en) | Bionic nano-carrier and application thereof in preparing medicament for treating brain glioma | |
| CN105012271A (en) | Doxorubicin and TRAIL co-supported albumin nanoparticle targeting preparation and preparation method thereof | |
| CN102961345B (en) | Method for preparing rapamycin/magnetic carboxymethyl chitosan nano drug-loaded microspheres | |
| CN104940945A (en) | Hollow mesoporous copper sulfide compound modified by hyaluronic acid and preparation method and application thereof | |
| Lin et al. | Development of a 5-fluorouracil-loaded PLGA microsphere delivery system by a solid-in-oil-in-hydrophilic oil (S/O/hO) novel method for the treatment of tumors | |
| CN104352442B (en) | A kind of mifepristone chitosan sustained-release microsphere preparation and preparation method thereof | |
| CN104288093B (en) | Application of the nano drug transdermal preparation in tumour | |
| Chen et al. | Macrophage-derived biomimetic nanoparticles enhanced SDT combined with immunotherapy inhibited tumor growth and metastasis | |
| CN104434792A (en) | Polymer micelle, preparation method thereof, antitumor pharmaceutical composition, preparation and preparation method thereof | |
| CN107308461A (en) | A kind of nano-particle of colon tumor cell active targeting and preparation method thereof | |
| CN110711249B (en) | Preparation method of lysosome membrane-coated nanoparticles | |
| CN107335064A (en) | A kind of nano-particle for macrophage targeted delivery of drugs and preparation method thereof | |
| CN107115320A (en) | A kind of targeted nano granule for loading Temozolomide and preparation method thereof | |
| CN107812189B (en) | Hypocrellin nano preparation for actively targeting specific tumor cells and preparation method and application thereof | |
| CN116440265A (en) | Targeted long-acting heat-generating nano-composite for enhancing photothermal treatment of tumor and application thereof | |
| Song et al. | Preparation and evaluation of insulin-loaded nanoparticles based on hydroxypropyl-β-cyclodextrin modified carboxymethyl chitosan for oral delivery | |
| CN115089561A (en) | Cell membrane coated prodrug nanoparticle, preparation method and application | |
| CN107970242A (en) | A kind of mesoporous silicon oxide of paclitaxel loaded/Tarceva-hyaluronic acid mixing targeting nano particle | |
| CN113827724A (en) | Drug-loaded Prussian blue @ manganese fibrin composite gel, and preparation method and application thereof | |
| CN102078302B (en) | Method for preparing berbamine chitosan-agar nano particles | |
| CN102846553A (en) | Preparation method for chlorella polypeptide-chitosan nanoparticle |
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 | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20171103 |
|
| WD01 | Invention patent application deemed withdrawn after publication |