CN103536919A - Tumor-targeted photodynamic medicine carrying nanoparticle as well as preparation method and application thereof - Google Patents
Tumor-targeted photodynamic medicine carrying nanoparticle as well as preparation method and application thereof Download PDFInfo
- Publication number
- CN103536919A CN103536919A CN201310505945.4A CN201310505945A CN103536919A CN 103536919 A CN103536919 A CN 103536919A CN 201310505945 A CN201310505945 A CN 201310505945A CN 103536919 A CN103536919 A CN 103536919A
- Authority
- CN
- China
- Prior art keywords
- porphyrin
- medicine
- nano particles
- light power
- preparation
- 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.)
- Granted
Links
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a tumor-targeted photodynamic medicine carrying nanoparticle as well as a preparation method and an application thereof, belonging to a medical preparation containing porphyrin or a medical preparation prepared by a method which utilizes wave energy to process materials. According to the invention, porphyrin or derivates of porphyrin of a functional group and polyethylene glycol and polypropylene glycol of a polyether chain segment are connected through chemical bonds or ester bonds or amido bonds of perssads, and then polymerized into a nanoparticle aqueous dispersion of porphyrin or derivates, loaded with a chemotherapy drug with a conjugated structure and dialyzed to obtain the tumor-targeted photodynamic medicine carrying nanoparticle. The nano material prepared by the method is high in yield, regular in shape, uniform in distribution and good in biosecurity, can be effectively loaded with an antitumor drug with a conjugated structure; a water-soluble photosensitizer can effectively reverse drug tolerance of a chemotherapy drug after being irradiated by near-infrared light; the nanoparticle can efficiently target and position tumors, has good anelasticity, effectively inhibits tumor growth, and has a wide application prospect in preparing breast cancer tumor-targeted drugs.
Description
Technical field
The present invention relates to a kind of pharmaceutical product that contains porphyrin, or utilize wave energy to process the medicinal preparation that material makes, be specifically related to light power medicine-carried nano particles of a kind of cancer target and its production and use.
Background technology
Chemotherapy is one of important means for the treatment of malignant tumor, yet a large amount of clinical practices shows that the medication effect of tumor is unsatisfactory, and its reason mainly comprises: (1) chemotherapeutics is poor to tumor cells selectivity; The current chemotherapeutics chemotherapeutics for oncotherapy, when killing cancerous cell, also damages patient's normal cell, and toxic and side effects is large, has greatly limited dosage, has a strong impact on therapeutic effect.(2) multidrug resistant of tumor cell mechanism; Intrinsic high expressed ABC albumen (as P-glycoprotein) on tumor cell membrane, can by various antitumor drug from intracellular transport to born of the same parents, cause drug level in born of the same parents to reduce, show natural resistant characterization, and in medication process, can also induce the expression of these ABC albumen, thereby generation acquired drug-resistance, has further reduced the therapeutic effect of medicine.Up to now, for hepatocarcinoma, there is no effective chemotherapeutics.Therefore, strengthening the tumor-targeting of chemotherapeutics and improve medicine is the important channel of improving chemotherapy of tumors effect to the chemosensitivity of tumor cell (improving drug effect).
The active platform that multifunctional nano-carrier can transmit as multi-medicament, by unique surface signal and the controlled orderly release to medicine of identification targeted cells, make medicine performance synergistic function, and significantly reduce the toxic and side effects of medicine, aspect chemotherapy of tumors, there is unique advantage.The existing large quantity research of delivering in medicine cancer target about Application of micron at present, although obtained certain effect, due to its biocompatibility, has finally all greatly limited its clinical practice.Therefore, utilize the own material of organism to prepare tumour medicine carrier, realize the targeting Delivery of tumor, become the key issue that solves chemotherapy of tumors treatment.
Photodynamic therapy (photo-dynamic therapy, PDT) be a kind of new method for the treatment of tumor that twentieth century the 80's new development is got up, photosensitive drug enters in body, be enriched in after focus, under the laser action of coupling absorbing wavelength, can generate active very strong singlet oxygen, produce cytotoxic effect, and then cause cell impaired and even dead.Because photodynamic therapy all has alternative to target tissue and degree of injury, compare with conventional therapy means such as operation, chemotherapy, radiotherapies, can reduce the damage of normal tissue.Current research shows: optical dynamic therapy can Chemotherapy, and photodynamic therapy provides new approaches for oncotherapy.
Although optical dynamic therapy has a lot of advantages in the treatment of tumor, also face a lot of problems: 1) most photosensitizer is all hydrophobic, need to, with using after certain organic solvent dissolution, therefore be difficult to be injected directly in patient's body; 2) photosensitizer metabolism is in vivo very fast, is therefore difficult to effectively assemble at lesions position; 3) it is very limited that optical dynamic therapy self suppresses tumor effect.Therefore, photodynamic therapy in oncotherapy usually used as the supplemental treatment regimens of tumor.
Summary of the invention
The present invention is exactly the chemotherapy resistance problem that chemotherapeutics targeting stationkeeping ability is poor in order to overcome, long-term prescription produces, and a kind of light power medicine-carried nano particles and preparation method and purposes that is directly used in the cancer target of clinical injection application is provided.
Design principle of the present invention: build nanometer medicine-carried system and deliver to realize the cancer target of medicine, and improve chemotherapy effect in conjunction with photodynamic therapy, realize chemotherapy sensitizing.Designed and take the own material of organism---porphyrin is prepared nano-medicament carrier as raw material, the macro ring conjugated structure forming due to porphyrin self, with the chemotherapeutics that contains conjugated structure by π-pi-conjugated high-efficient carrier medicine; Porphyrin can optionally be gathered in tumor tissues as photosensitizer, can realize the targeting Delivery of medicine after carrying medicament.Therefore, porphyrin material all has a good application prospect in fields such as medical science and material science.Although the drug targeting in tumor is controlled in delivery and photo-dynamical medicine therapeutic process and obtained certain achievement in research, all there are some problems, and there is no so far a kind of technology that water miscible photosensitizer is directly used in to targeted drug delivery.
Be the present invention take porphyrin or derivatives thereof and functionalization group end capping polyethers as raw material synthetic, in hemoporphyrin structure, there are two carboxyls and two hydroxyls, improved water solublity, and utilize reactivity and the polyether grafting of carboxyl and/or hydroxyl, the synthetic polymer of preparing nanoparticle, and prepare by dialysis.
The present invention realizes according to following technical scheme.
A kind of light power medicine-carried nano particles of cancer target, comprise the polymer of being prepared by polyether segment and functional group, it is characterized in that: the porphyrin or derivatives thereof of described functional group and the Polyethylene Glycol of polyether segment, ester bond or amido link through chemical bond or group between polypropylene glycol connect, make porphyrin or derivatives thereof nanoparticle aqueous dispersions, and carry the chemotherapeutics with conjugated structure and form, porphyrin ring shines and excites lower generation singlet oxygen near infrared light, suppress tumor growth, the particle diameter of described nanoparticle is 20-80nm, profile is spherical or subglobular.
The preparation method of light power medicine-carried nano particles,
1. porphyrin or derivatives thereof-Polyethylene Glycol nano-medicament carrier is synthetic
A. press 0.5-3mg/1ml concentration by the mixed solution stirred overnight at room temperature of porphyrin or derivatives thereof and MES (MES);
B. by the EDC 3-20 mg of activated carboxylic group, NHS 2-15mg room temperature activation 5-6h, add in above-mentioned mixed solution 10ml, under room temperature, stir after 2 ~ 8h, add 2 mercapto ethanol 15-85 ul cancellation EDC, NaOH adjusts pH value to 7.5 ~ 8.0, standby;
C. solution after cancellation is added drop-wise to slowly to concentration and is in the phosphate buffer 1 0ml of isopropyl Polyethylene Glycol of 4-10mg/1ml double-end amino end-blocking, stirring is spent the night, and be transferred in bag filter, 48h dialyses in borate buffer solution, obtain porphyrin or derivatives thereof nanoparticle aqueous dispersions, standby;
2. medicine carrying porphyrin or derivatives thereof nano material is synthetic
A. by concentration 1-2mg/1ml, will there is conjugated structure chemotherapeutics and be dissolved in distilled water, under room temperature, stir 1h, standby;
B. press 0.5mg/ml concentration, get the above-mentioned medicinal liquid of 2ml and dropwise join in 1-8 mL porphyrin or derivatives thereof nanoparticle aqueous dispersions, 4 ℃ of room temperature lucifuges are placed in bag filter after spending the night, distilled water dialysis 2h, obtaining particle diameter is the light power medicine-carried nano particles of the cancer target of 20-80nm.
The application of the light power medicine-carried nano particles of described cancer target in preparation treatment cancer drug,
A. the application of cancer target light power medicine-carried nano particles in preparation treatment breast cancer medicines;
B. the application of cancer target light power medicine-carried nano particles in preparing Hepatoma therapy medicine.
The present invention has the following advantages:
1. carrier medicine carrying efficiency is high, and carrying drug ratio reaches more than 80%, and has good slow release and controlled release ability;
2. after nano material near infrared light, effectively assist chemotherapeutics amycin to enter nucleus from endochylema;
3. medicine half-inhibition concentration reduces more than 8 times, has effectively reversed chemotherapeutics tolerance;
4. after near infrared light nano-medicament carrier, effectively start mitochondrion-apoptotic signal path, cell death inducing;
5. injection for curing in tail vein body, the efficient targeting positioning tumor of nano-medicament carrier, and have good anelasticity and effectively suppress tumor growth.
Therefore, the present invention has initiatively targeting to tumor, can efficiently carry the antitumor drug with conjugated structure, realizes medicine in the orderly release at tumor focus position, and the inside and outside growth according to effective inhibition tumor under responding near infrared light.
Accompanying drawing explanation
Fig. 1 is the nuclear magnetic spectrum of raw material hemoporphyrin (HP);
Fig. 2 is the nuclear magnetic spectrum of double-end amino end-blocking isopropyl Polyethylene Glycol (PEG);
Fig. 3 is the nuclear magnetic spectrum of synthetic hemoporphyrin-Polyethylene Glycol nano-medicament carrier (HPP);
Fig. 4 be in embodiment 1 HP atomic force microscopy;
Fig. 5 is the atomic force microscopy of HPP in embodiment 1;
Fig. 6 is the transmission electron microscope photo of HP in embodiment 1;
Fig. 7 is the transmission electron microscope photo of HPP in embodiment 1;
Fig. 8 is the raw material hemoporphyrin of different proportion in embodiment 1, the fluorogram of amycin (Dox, ADR) synthesizing nano-particle;
Fig. 9 is that the HPPD nanoparticle of preparation in embodiment 1 is at different pH value and have or not the drug release curve under near-infrared illumination condition;
The particle size distribution figure of the HPPD nanoparticle of preparation in Figure 10 embodiment 1;
Figure 11 is that the HPPD nanoparticle of preparation in embodiment 1 is measured curve chart to Adriamycin resistant breast cancer cell (ADR/MCF-7) medicine half-inhibition concentration;
Figure 12 be in embodiment 1 preparation HPPD nanoparticle to tumor killing effect in the body of ADR/MCF-7 mouse model and tissue distribution figure;
Figure 13 is the tissue distribution figure of illumination HPPD nanoparticle to ADR/MCF-7 mouse model not in embodiment 1;
Figure 14 is the tissue distribution figure of illumination HPPD nanoparticle to ADR/MCF-7 mouse model in embodiment 1.
The specific embodiment
Below in conjunction with drawings and Examples, the present invention is specifically described.
A kind of light power medicine-carried nano particles of cancer target, comprise the polymer of being prepared by polyether segment and functional group, ester bond or amido link through chemical bond or group between the porphyrin or derivatives thereof of described functional group and the Polyethylene Glycol of polyether segment, polypropylene glycol are connected, make porphyrin or derivatives thereof nanoparticle aqueous dispersions, and carry the chemotherapeutics with conjugated structure and form, porphyrin ring shines and excites lower generation singlet oxygen near infrared light, suppress tumor growth, the particle diameter of described nanoparticle is 20-80nm, and profile is spherical or subglobular.
The light power medicine-carried nano particles of described cancer target, is rufous polymer powder after its porphyrin or derivatives thereof-Polyethylene Glycol nanoparticle aqueous dispersions lyophilization, or purple polymer powder.
The preparation method of light power medicine-carried nano particles,
1. porphyrin or derivatives thereof-Polyethylene Glycol nano-medicament carrier is synthetic
A. press 0.5-5mg/1ml concentration by the mixed solution stirred overnight at room temperature of porphyrin or derivatives thereof and MES (MES);
B. by the 1-ethyl of activated carboxylic group-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) 3-20 mg, N-hydroxy-succinamide (NHS) 2-15 mg room temperature activation 5-6h, add in above-mentioned mixed solution 10ml, under room temperature, stir after 2 ~ 8h, add 2 mercapto ethanol 15-85 ul cancellation EDC, NaOH adjusts pH value to 7.5 ~ 8.0, standby;
C. solution after cancellation is added drop-wise to slowly to concentration and is in phosphate buffer (PBS) 10ml of isopropyl Polyethylene Glycol of 4-10mg/1ml double-end amino end-blocking, stirring is spent the night, and be transferred in bag filter, 48h dialyses in borate buffer solution, obtain porphyrin or derivatives thereof nanoparticle aqueous dispersions, standby;
2. medicine carrying porphyrin or derivatives thereof nano material is synthetic
A. by concentration 1-2mg/1ml, will there is conjugated structure chemotherapeutics and be dissolved in distilled water, under room temperature, stir 1h, standby;
B. press 0.5mg/ml concentration, get the above-mentioned medicinal liquid of 2ml and dropwise join in 1-8 mL porphyrin or derivatives thereof nanoparticle aqueous dispersions, 4 ℃ of room temperature lucifuges are placed in bag filter after spending the night, distilled water dialysis 2h, obtaining particle diameter is the light power medicine-carried nano particles of the cancer target of 20-80nm.
The preparation method of the light power medicine-carried nano particles of described tumor targeting, it has conjugated structure chemotherapeutics is amycin, or epirubicin, or mitoxantrone.
The preparation method of the light power medicine-carried nano particles of described tumor targeting, described porphyrin or derivatives thereof is protoporphyrin, or hemoporphyrin, or uroporphyrin or coproporphyrin.
The preparation method of the light power medicine-carried nano particles of described tumor targeting, the molecular weight of the isopropyl Polyethylene Glycol of its double-end amino end-blocking is 2000-6000Da.
The preparation method of the light power medicine-carried nano particles of described tumor targeting, described in there is conjugated structure chemotherapeutics and the mass ratio in nano-medicament carrier be 8:1 ~ 1:8.
The preparation method of the light power medicine-carried nano particles of described tumor targeting, in the EDC of its activated carboxylic group, NHS and porphyrin or derivatives thereof, the mol ratio of carboxyl unit is 2:1 ~ 1:1.
The preparation method of the light power medicine-carried nano particles of described tumor targeting, the molecular cut off of its bag filter as described in 2. b is 2000-6000Da, is greater than the molecular weight polyethylene glycol of use simultaneously; After dialysis, the particle diameter of medicine-carried nano particles is 20-80nm.
The application of the light power medicine-carried nano particles of described cancer target in preparation treatment cancer drug,
A. the application of cancer target light power medicine-carried nano particles in preparation treatment breast cancer medicines;
B. the application of cancer target light power medicine-carried nano particles in preparing Hepatoma therapy medicine.
Embodiment mono-:
1, hemoporphyrin-Polyethylene Glycol (HPP) nano-medicament carrier is synthetic
2, medicine carrying hemoporphyrin (HPPD) nano material is synthetic
Get 20mg amycin (Dox) and be dissolved in 10mL distilled water, under room temperature, stir 1h, standby; Get the amycin solution that 2mL prepares and dropwise join 5mL HPP nanoparticle aqueous dispersions (0.5mg/mL), 4 ℃ spend the night after, be placed in bag filter, with distilled water dialysis treatment 2h.Fig. 4-7 are shown in by its structural identification collection of illustrative plates.Particle size distribution is shown in Figure 10.The drug loading of amycin adopts ultraviolet spectrophotometer method to detect (detection wavelength is 480nm), is 85.2%, the results are shown in Figure 8.Medicament-carried nano material particle size 35nm.
3, the vitro drug release of medicine-carried nano particles
10mL HPPD is put in bag filter (molecular cut off 1000 Da), be placed in respectively the phosphate buffered solution (PBS) of 50mL pH 5.8 and pH 7.4,37 ℃, 100 rpm lucifuge vibrations, the stipulated time whole release medium of naming a person for a particular job is replaced by fresh medium, and ultraviolet spectrophotometer method detects (detection wavelength is 480nm).Drug release percentage rate calculates according to following formula: discharge percentage rate=(release amount of medicine/medicine total amount) * 100%.Result as shown in Figure 9, can effectively realize medicine and control release by medicine-carried nano particles.
4, the cytotoxicity of HPPD to drug resistance breast cancer cell ADR/MCF-7
By exponential phase of growth ADR/MCF-7 cell with 6000/every hole, be inoculated in 96 well culture plates, put 37 ℃, in 5% CO2 incubator, cultivate 48 h, the culture medium of inclining, add the free ADR of 150 μ L culture medium and variable concentrations or the HPPD solution under HPPD nano dispersion fluid and illumination condition, continue to cultivate 48 h.Adopt mtt assay: every hole adds 3-(4,5-dimethylthiazole-2)-2,5-diphenyl tetrazole bromine salt (MTT) 20 μ L, 37 ℃ are continued to hatch 4 h, stop cultivating.Culture supernatant in hole is abandoned in suction, and every hole adds dimethyl sulfoxide (DMSO) 150 μ L, and 10 min that vibrate select 490 nm wavelength, measure each hole absorbance value on enzyme-linked immunosorbent assay instrument, with blank zeroing, and establishes contrast non-processor hole.Survey the optical density value (OD value) in every hole, calculate survival rate;
Survival rate=(experimental port OD value/control wells OD value) * 100 %
According to cell survival rate (as Figure 11), utilize IC50 software for calculation to obtain IC50.Result demonstration, HPPD irradiates IC50 value and has reduced nearly 9 times compared with single therapy treatment group, effectively reduces drug dose, has reversed Chemoresistance.
5, HPPD accumulating and distributing in ADR/MCF-7 cell
Take 10.9 mg Fluorescein isothiocyanates (FITC) and be dissolved in 1mL DMSO, slowly add above-mentioned HPPD aqueous dispersions stirring reaction to spend the night, 48h is standby in dialysis.
After ADR/MCF-7 cell culture, be inoculated in glass bottom Tissue Culture Dish, cell concentration is 5 * 10
4cell/mL,
The HPPD nano dispersion fluid (ADR solubility 16.7 μ g/ml) that adds respectively free ADR and FITC labelling, 37 ℃ hatch 4 h after near infrared light according to 70s, after 24h, discard culture fluid, with cold PBS liquid, wash 3 times, 3.7% paraformaldehyde is fixed 15 min, with confocal laser scanning microscope (Figure 12).Result shows that HPPD can carry amycin medicine and enter nucleus by endochylema after infrared radiation, is conducive to reverse multiple drug resistance of tumor.
6, tumor killing effect in the body of HPPD
ADR/MCF-7 breast carcinoma mouse model: get nude mice (female), 4 ~ 6 week age, body weight 18 ~ 22g, be divided at random 5 groups, be respectively matched group, HPP+ irradiation group, free amycin (Dox) group, HPPD group and HPPD+ irradiation group, every group of 8 mices, by 3.2 mg/kg drug dose tail intravenously administrables, be administered three times weekly, administration three weeks.Measure gross tumor volume, draw tumor growth curve.Result shows, free drug group is undesirable to tumor suppression effect, finishes to the observation period, and gross tumor volume is growth continuously and healthily always, and relative tumour volume increases to 5.0 than (gross tumor volume and initial tumor volume ratio after treatment) from 1.0; And HPPD+ irradiation group can significantly suppress tumor growth, the 10th day observation period, can suppress tumor growth completely.
MHCC-97H liver cancer mouse model: get nude mice (female), 4 ~ 6 week age, body weight 18 ~ 22g, be divided at random 5 groups, be respectively matched group, free Dox group (5mg/kg), HPPD group (1.5mg/kg), HPPD group (5mg/kg), be administered three times weekly, two weeks.Measure gross tumor volume, draw tumor growth curve.Result shows, HPPD(1.5mg/kg) administration group is within the observation period, and relative tumour volume maintains 1.0 left and right all the time than (gross tumor volume and initial tumor volume ratio after treatment); HPPD(5mg/kg) after administration group two weeks, relative tumour volume is than having dropped to 0.2 by 1, and result shows, HPPD Nano medication can effectively suppress the tumor growth of tumor.
7, the tissue distribution of HPPD in Mice Body
The cy-5.5 that gets N-maloyl imines end-blocking end-blocking is dissolved in 20 μ L(1 mg/mL in DMSO), dropwise add the synthetic HPP of 500 μ L, stirring at room 2h, dialysis 48h(molecular cut off 1000 Da) after, preparation cy5.5-HPPD.
MHCC-97H liver cancer model: nude mice is divided into 3 groups at random, be respectively PBS-cy5.5, PEG-cy5.5, HPPD-cy5.5, press 0.4ug/g fluorescence dosage tail intravenously administrable, 2h and 24 h use small animal imaging instrument to observe Nano medication and respectively organize the distribution in internal organs mice.Result shows, after 2h tail vein injection PBS-cy5.5, fluorescence signal is strong, and is evenly distributed in mice and respectively organizes in internal organs, and after 24h, fluorescence all disappears substantially in respectively organizing internal organs, shows that cy5.5 fluorochrome does not have tissue-targeting; After PEG-cy5.5 group 2h tail vein injection, fluorescence concentrates in tumor and kidney, and 24h mainly concentrates on kidney; After HPPD-cy5.5 group 2h tail vein injection, fluorescence concentrates in tumor and kidney, and after 24h, fluorescence is mainly enriched in tumor, illustrates that Nano medication has very strong tumor-targeting and tumor anelasticity.
ADR/MCF-7 breast cancer model: nude mice is divided into 2 groups at random, be respectively HPPD-cy5.5, HPPD-cy5.5+ irradiation group, by 0.4 μ g/g fluorescence dosage tail intravenously administrable, 2h and 24 h use small animal imaging instrument to observe Nano medication and respectively organize the distribution in internal organs mice.Result shows, with respect to HPPD-cy5.5 group, irradiation group can more effectively increase Nano medication in the enrichment of tumor locus, reduces the picked-up of kidney.
Drug distribution of the present invention is shown in Figure 13,14.
8, the acute toxicity testing of HPPD in Mice Body.
Get nude mice (female), 4 ~ 6 week age, body weight 18 ~ 22 g, be divided at random 3 groups, be respectively matched group, free Dox group, HPPD, every group of 3 mices, press 10mg/kg drug dose tail intravenously administrable, after 24h, core, the organ such as liver, spleen, lung, kidney, tumor, after homogenate, add the amycin in methanol solution extraction tissue, 4 degree, the centrifugal 10min of 5000rpm, 2 times, get supernatant, ultraviolet spectrophotometer is measured the drug distribution of amycin in each organ.
Result shows, Dox carries by HPPD's, and the distribution of medicine in heart significantly reduces (from 84 μ g/g, being reduced to 60 μ g/g), is conducive to alleviate the cardiac toxicity of ADR; In addition,, by carrying of HPPD, Dox is mainly distributed in tumor tissues (50 are increased to 135 μ g/g), apparently higher than other organ-tissue.
Embodiment bis-:
Synthesizing of hemoporphyrin-Polyethylene Glycol (HPP) nano-medicament carrier
Embodiment tri-:
Synthesizing of hemoporphyrin-Polyethylene Glycol (HPP) nano-medicament carrier
Embodiment tetra-:
Synthesizing of coproporphyrin-Polyethylene Glycol (PP) nano-medicament carrier
Embodiment five:
Synthesizing of medicine carrying hemoporphyrin (HPPD) nano material
Get 20mg epirubicin and be dissolved in 10mL distilled water, under room temperature, stir 1h, standby; Get the HPP nanoparticle aqueous dispersions (0.5mg/mL) that epirubicin solution that 1mL prepares dropwise joins preparation in 0.5mL embodiment 1, wherein, the mass ratio of chemotherapeutics and nano-medicament carrier is 8:1, after 4 ° of C spend the night, be placed in bag filter, with distilled water dialysis treatment 2h.Drug loading adopts ultraviolet spectrophotometer method to detect, and is 80.1%.Dynamic light scattering records particle diameter 67nm.
Embodiment six:
Synthesizing of medicine carrying hemoporphyrin (HPPD) nano material
Get 10mg epirubicin and be dissolved in 10mL distilled water, under room temperature, stir 1h, standby; Get the HPP nanoparticle aqueous dispersions (0.5mg/mL) that epirubicin solution that 1mL prepares dropwise joins preparation in 2mL embodiment 1, wherein, the mass ratio of chemotherapeutics and nano-medicament carrier is 1:1, after 4 ° of C spend the night, be placed in bag filter, with distilled water dialysis treatment 2h.Drug loading adopts ultraviolet spectrophotometer method to detect, and is 86.1%.Dynamic light scattering records particle diameter 70nm.
Synthesizing of medicine carrying hemoporphyrin (HPPD) nano material
Get 5mg epirubicin and be dissolved in 10mL distilled water, under room temperature, stir 1h, standby; Get the HPP nanoparticle aqueous dispersions (0.5mg/mL) that epirubicin solution that 1mL prepares dropwise joins preparation in 8mL embodiment 1, wherein, the mass ratio of chemotherapeutics and nano-medicament carrier is 1:8, after 4 ° of C spend the night, be placed in bag filter, with distilled water dialysis treatment 2h.Drug loading adopts ultraviolet spectrophotometer method to detect, and is 89.3%.Dynamic light scattering records particle diameter 73nm.
Embodiment seven:
Synthesizing of medicine carrying hemoporphyrin (HPPD) nano material
Get 10mg mitoxantrone and be dissolved in 10mL distilled water, under room temperature, stir 1h, standby; Get the HPP nanoparticle aqueous dispersions (0.5mg/mL) that mitoxantron solutions that 2mL prepares dropwise joins preparation in 5mL embodiment 1, wherein, the mass ratio of chemotherapeutics and nano-medicament carrier is 4:5, after 4 ° of C spend the night, be placed in bag filter, with distilled water dialysis treatment 2h.Drug loading adopts ultraviolet spectrophotometer method to detect, and is 85.2%.Dynamic light scattering records particle diameter 62nm.
In the vitro drug release of the medicine-carried nano particles of embodiment five and embodiment six, cytotoxicity, body, in the experiment such as tumor killing effect and embodiment 1, data are similar, at this repeated description not.
Claims (10)
1. the light power medicine-carried nano particles of a cancer target, comprise the polymer of being prepared by polyether segment and functional group, it is characterized in that: the porphyrin or derivatives thereof of described functional group and the Polyethylene Glycol of polyether segment, ester bond or amido link through chemical bond or group between polypropylene glycol connect, make porphyrin or derivatives thereof nanoparticle aqueous dispersions, and carry the chemotherapeutics with conjugated structure and form, porphyrin ring shines and excites lower generation singlet oxygen near infrared light, suppress tumor growth, the particle diameter of described nanoparticle is 20-80nm, profile is spherical or subglobular.
2. the light power medicine-carried nano particles of cancer target according to claim 1, is characterized in that: after the lyophilization of porphyrin or derivatives thereof-Polyethylene Glycol nanoparticle aqueous dispersions, be rufous powdery product, or purple powdery product.
3. the preparation method of the light power medicine-carried nano particles of cancer target as claimed in claim 1, is characterized in that:
1. porphyrin or derivatives thereof-Polyethylene Glycol nano-medicament carrier is synthetic
A. press 0.5-3mg/1ml concentration by the mixed solution stirred overnight at room temperature of porphyrin or derivatives thereof and MES (MES);
B. by the EDC 3-20 mg of activated carboxylic group, NHS 2-15mg room temperature activation 5-6h, add in above-mentioned mixed solution 10ml, under room temperature, stir after 2 ~ 8h, add 2 mercapto ethanol 15-85 ul cancellation EDC, NaOH adjusts pH value to 7.5 ~ 8.0, standby;
C. solution after cancellation is added drop-wise to slowly to concentration and is in the phosphate buffer 1 0ml of isopropyl Polyethylene Glycol of 4-10mg/1ml double-end amino end-blocking, stirring is spent the night, and be transferred in bag filter, 48h dialyses in borate buffer solution, obtain porphyrin or derivatives thereof nanoparticle aqueous dispersions, standby;
2. medicine carrying porphyrin or derivatives thereof nano material is synthetic
A. by concentration 1-2mg/1ml, will there is conjugated structure chemotherapeutics and be dissolved in distilled water, under room temperature, stir 1h, standby;
B. press 0.5mg/ml concentration, get the above-mentioned medicinal liquid of 2ml and dropwise join in 1-8 mL porphyrin or derivatives thereof nanoparticle aqueous dispersions, 4 ℃ of room temperature lucifuges are placed in bag filter after spending the night, distilled water dialysis 2h, obtaining particle diameter is the light power medicine-carried nano particles of the cancer target of 20-80nm.
4. the preparation method of the light power medicine-carried nano particles of tumor targeting according to claim 3, is characterized in that: having conjugated structure chemotherapeutics is amycin, or epirubicin, or mitoxantrone.
5. the preparation method of the light power medicine-carried nano particles of tumor targeting according to claim 3, is characterized in that: described porphyrin or derivatives thereof is protoporphyrin, or hemoporphyrin, or uroporphyrin or coproporphyrin.
6. the preparation method of the light power medicine-carried nano particles of tumor targeting according to claim 3, is characterized in that: the molecular weight of the isopropyl Polyethylene Glycol of double-end amino end-blocking is 1500-5000Da.
7. the preparation method of the light power medicine-carried nano particles of tumor targeting according to claim 3, is characterized in that: described in there is conjugated structure chemotherapeutics and the mass ratio in nano-medicament carrier be 8:1 ~ 1:8.
8. the preparation method of the light power medicine-carried nano particles of tumor targeting according to claim 3, is characterized in that: in the EDC of activated carboxylic group, NHS and porphyrin or derivatives thereof, the mol ratio of carboxyl unit is 2:1 ~ 1:1.
9. the preparation method of the light power medicine-carried nano particles of tumor targeting according to claim 3, is characterized in that: as described in 2. b, the molecular cut off of bag filter is 2000-6000Da, is greater than the molecular weight polyethylene glycol of use simultaneously; After dialysis, the particle diameter of medicine-carried nano particles is 20-80nm.
10. the application of the light power medicine-carried nano particles of cancer target in preparation treatment cancer drug as described in claim 1 or 3,
A. the application of cancer target light power medicine-carried nano particles in preparation treatment breast cancer medicines;
B. the application of cancer target light power medicine-carried nano particles in preparing Hepatoma therapy medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310505945.4A CN103536919B (en) | 2013-10-24 | 2013-10-24 | Tumor-targeted photodynamic medicine carrying nanoparticle as well as preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310505945.4A CN103536919B (en) | 2013-10-24 | 2013-10-24 | Tumor-targeted photodynamic medicine carrying nanoparticle as well as preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103536919A true CN103536919A (en) | 2014-01-29 |
| CN103536919B CN103536919B (en) | 2015-07-15 |
Family
ID=49961028
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310505945.4A Active CN103536919B (en) | 2013-10-24 | 2013-10-24 | Tumor-targeted photodynamic medicine carrying nanoparticle as well as preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103536919B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105833289A (en) * | 2016-05-30 | 2016-08-10 | 上海交通大学 | Mitochondrion-targeting nano-drug delivery system and preparation method and application thereof |
| CN106551904A (en) * | 2015-09-18 | 2017-04-05 | 天津医科大学 | The sound of target tumor, light power medicament-carried nano micelle and its production and use |
| CN108619510A (en) * | 2018-04-02 | 2018-10-09 | 东南大学 | A kind of synthetic method of EPS-RB nano particles for light power antibacterial |
| CN110702899A (en) * | 2019-10-12 | 2020-01-17 | 香港大德昌龙生物科技有限公司 | Biotin labeled DNA, preparation method and kit thereof |
| CN113773488A (en) * | 2021-09-06 | 2021-12-10 | 大连理工大学 | Preparation method of water-soluble polyporphyrin carrier-free nano-drug |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102573914A (en) * | 2009-10-16 | 2012-07-11 | 大学健康网络 | Porphyrin nanovesicles |
| CN102940892A (en) * | 2012-11-21 | 2013-02-27 | 哈尔滨工业大学 | Preparation method of targeting porphyrin fluorescent molecule and gold nanorod dyad |
| CN103239718A (en) * | 2013-05-13 | 2013-08-14 | 江苏大学 | Method for preparing adriamycin-loaded polycaprolactone-block-polyethylene glycol nano microspheres |
-
2013
- 2013-10-24 CN CN201310505945.4A patent/CN103536919B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102573914A (en) * | 2009-10-16 | 2012-07-11 | 大学健康网络 | Porphyrin nanovesicles |
| CN102940892A (en) * | 2012-11-21 | 2013-02-27 | 哈尔滨工业大学 | Preparation method of targeting porphyrin fluorescent molecule and gold nanorod dyad |
| CN103239718A (en) * | 2013-05-13 | 2013-08-14 | 江苏大学 | Method for preparing adriamycin-loaded polycaprolactone-block-polyethylene glycol nano microspheres |
Non-Patent Citations (4)
| Title |
|---|
| LUC BARRAUD ET AL.: "Increase of doxorubicin sensitivity by doxorubicin-loading into nanoparticles for hepatocellular carcinoma cells in vitro and in vivo", 《JOURNAL OF HEPATOLOGY》, vol. 42, 31 December 2005 (2005-12-31) * |
| RENE HORNUNG ET AL.: "Highly Selective Targeting of Ovarian Cancer with the Photosensitizer PEG-m-THPC in a Rat Model", 《PHOTOCHEMISTRY AND PHOTOBIOLOGY》, vol. 70, no. 4, 31 December 1999 (1999-12-31) * |
| S. K. SAHOO ET AL.: "Pegylated Zinc Protoporphyrin: A Water-Soluble Heme Oxygenase Inhibitor with Tumor-Targeting Capacity", 《BIOCONJUGATE CHEM.》, vol. 13, 31 December 2002 (2002-12-31) * |
| WUBEANTE YENET AYEN ET AL.: "Doxorubicin-Loaded (PEG)3-PLA Nanopolymersomes: Effect of Solvents and Process Parameters on Formulation Development and In Vitro Study", 《MOL. PHARMACEUTICS》, vol. 8, 2 February 2011 (2011-02-02) * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106551904A (en) * | 2015-09-18 | 2017-04-05 | 天津医科大学 | The sound of target tumor, light power medicament-carried nano micelle and its production and use |
| CN105833289A (en) * | 2016-05-30 | 2016-08-10 | 上海交通大学 | Mitochondrion-targeting nano-drug delivery system and preparation method and application thereof |
| CN105833289B (en) * | 2016-05-30 | 2019-04-09 | 上海交通大学 | A kind of Mitochondrially targeted Nano medication delivery system and the preparation method and application thereof |
| CN108619510A (en) * | 2018-04-02 | 2018-10-09 | 东南大学 | A kind of synthetic method of EPS-RB nano particles for light power antibacterial |
| CN110702899A (en) * | 2019-10-12 | 2020-01-17 | 香港大德昌龙生物科技有限公司 | Biotin labeled DNA, preparation method and kit thereof |
| CN113773488A (en) * | 2021-09-06 | 2021-12-10 | 大连理工大学 | Preparation method of water-soluble polyporphyrin carrier-free nano-drug |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103536919B (en) | 2015-07-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhao et al. | Nanotechnology for cancer therapy based on chemotherapy | |
| Sun et al. | One-pot fabrication of hollow porphyrinic MOF nanoparticles with ultrahigh drug loading toward controlled delivery and synergistic cancer therapy | |
| Zhang et al. | Ultrasound-enhanced chemo-photodynamic combination therapy by using albumin “nanoglue”-based nanotheranostics | |
| Huang et al. | Nanosonosensitizers for highly efficient sonodynamic cancer theranostics | |
| Tian et al. | Recent advances in upconversion nanoparticles‐based multifunctional nanocomposites for combined cancer therapy | |
| Cui et al. | In vivo targeted deep-tissue photodynamic therapy based on near-infrared light triggered upconversion nanoconstruct | |
| Zhang et al. | Carrier-free, chemophotodynamic dual nanodrugs via self-assembly for synergistic antitumor therapy | |
| Zou et al. | Phototherapy meets immunotherapy: A win–win strategy to fight against cancer | |
| Luo et al. | Conjugation of a scintillator complex and gold nanorods for dual-modal image-guided photothermal and X-ray-induced photodynamic therapy of tumors | |
| Choi et al. | Light-triggered photodynamic nanomedicines for overcoming localized therapeutic efficacy in cancer treatment | |
| CN108159422A (en) | A kind of preparation method of self assembly drug-loading system and its compound formulation | |
| Jin et al. | NIR-triggered sequentially responsive nanocarriers amplified cascade synergistic effect of chemo-photodynamic therapy with inspired antitumor immunity | |
| CN103536919B (en) | Tumor-targeted photodynamic medicine carrying nanoparticle as well as preparation method and application thereof | |
| Wang et al. | Biodegradable hollow polydopamine@ manganese dioxide as an oxygen self-supplied nanoplatform for boosting chemo-photodynamic cancer therapy | |
| Yue et al. | Melanin-like nanomedicine in photothermal therapy applications | |
| CN107007835B (en) | Prussian blue-loaded targeting nano-composite and preparation method thereof | |
| Li et al. | From one to all: self-assembled theranostic nanoparticles for tumor-targeted imaging and programmed photoactive therapy | |
| Bao et al. | Nanoscale metal–organic framework composites for phototherapy and synergistic therapy of cancer | |
| Hao et al. | Combinatorial therapeutic approaches with nanomaterial-based photodynamic cancer therapy | |
| Chen et al. | Dual-mode imaging and therapeutic effects of drug-loaded phase-transition nanoparticles combined with near-infrared laser and low-intensity ultrasound on ovarian cancer | |
| CN107875384A (en) | A kind of neoplasm targeted therapy drug delivery system for containing sensitising agent | |
| Luo et al. | Surface engineering of lanthanide nanoparticles for oncotherapy | |
| Geng et al. | GSH-sensitive nanoscale Mn3+-sealed coordination particles as activatable drug delivery systems for synergistic photodynamic-chemo therapy | |
| CN107638566A (en) | A kind of multifunctional nano capsule for integrating near-infrared fluorescence imaging and chemotherapy/photo-thermal therapy | |
| Dias et al. | Current photoactive molecules for targeted therapy of triple-negative breast cancer |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |