CN105561304A - Toad venom targeted liposome and preparation method and application thereof - Google Patents
Toad venom targeted liposome and preparation method and application thereof Download PDFInfo
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- Medicinal Preparation (AREA)
Abstract
The present invention discloses a cancer-treating toad venom targeted liposome and a preparation method and application thereof. The toad venom targeted liposome includes a targeted site carbonic anhydrase CA9 antibody, a liposome and toad venom. The liposome covers the toad venom, and then is connected with the carbonic anhydrase CA9 antibody, so that the toad venom targeted liposome can specifically act on CA9 overexpressing cancer cells for targeted killing of the cancer cells. Results of studies on characteristics of the prepared toad venom targeted liposome, tumor cell MTT experiments and in-vivo pharmacodynamic experiments show that the toad venom targeted liposome is relatively stable, difficult to gather, good in targeting property and high in therapeutic index. The toad venom targeted liposome is useful in the treatment of colon cancer, bladder cancer, renal cancer, cervical cancer, brain glioma and gastric cancer.
Description
Technical field
The invention belongs to field of pharmaceutical preparations, relate to target liposomes, particularly relate to the Chansu Liposome of CAC A9 antibody modification and construction method thereof and the application in antitumor thereof.
Background technology
Venenum Bufonis (Bufotoxin) is Bufo siccus (Toad, popular name toad) ear rear gland and the dry secretions of skin body of gland, energy heart tonifying, excited respiratory center and striped muscle, there is raising blood pressure, ease pain, relieving asthma, antitussive, antiinflammatory and antitumor action, be important Chinese medicine material.Venenum Bufonis enters ball more, falls apart, famous prepared Chinese traditional medicine liushen pill, pills for throat disease, Melnikov's method all contain this taste medicine of Venenum Bufonis, in addition, the Venenum Bufonis medicine that State Food and Drug Administration also ratifies to go on the market has troche of toad venom, Venenum Bufonis Injection, Venenum Bufonis analgesia ointment, compound recipe Chansu Wan, compound recipe toad venom plaster, Venenum Bufonis analgesia Babu cream.But Venenum Bufonis belongs to 2 class toxic traditional Chinese medicines, the highest consumption is 0.015-0.03g, contained bufotoxin is by excited vagus maincenter or tip, directly act on cardiac muscle, there is nausea,vomiting,diarrhea, suffer from abdominal pain, be short of physical strength, dizziness, cardiopalmus, the symptom such as uncomfortable in chest, lip and numb limbs and tense tendons, can be there is cyanotic lips, tic in severe patient, even occur circulation, respiratory system exhaustion and cause death.These toxic and side effects considerably increase the risk of Venenum Bufonis Clinical practice, so the improvement of clinical rational drug use, preparation process, seem particularly important to raising drug safety.
1993, scholar is had to find MN albumen in the HeLa cell line of the cervical cancer with human breast cancer cell Combined culture.Research subsequently finds, this protein structure is similar to carbonic anhydrase (CA), and is identified as a kind of isomerase of carbonic anhydrase, is then named as CA9.The transmembrane glycoprotein that it is made up of acidic amino acid, be made up of 459 aminoacid, be distributed in cell membrane and nucleus, relative molecular mass is respectively 58kD, 54kD.CAC A9 participates in the acid-base value regulating intraor extracellular, plays a significant role in the growth and survival course of tumor cell.In the ordinary course of things, CA9 is only expressed in few normal structure, as stomach, bile duct, pancreas, small intestinal and deferent duct epithelium, and expression is few, and can high expressed CAC A9 in colon cancer, bladder cancer, renal carcinoma, cervical cancer, cerebral glioma, gastric cancer, in prior art, major part is used to the diagnosis of tumor, although there is description CA9 can be used for the treatment of in prior art, do not provide and specifically how to utilize CA9 to treat.
Summary of the invention
1, object of the present invention.
The present invention effectively can utilize the property of medicine of Venenum Bufonis in prior art in order to solve, the toxic and side effects that the huge toxicity simultaneously reducing Venenum Bufonis medicine produces human body, and can suppress cancerous cell targetedly, reduce the injury to human normal cell and patient treatment misery, improve therapeutic effect, thus a kind of Venenum Bufonis target liposomes proposed and its preparation method and application.
2, the technical solution adopted in the present invention.
(1) Venenum Bufonis target liposomes
Venenum Bufonis target liposomes, draw together CAC A9 antibody, Venenum Bufonis and liposome, Venenum Bufonis is wrapped in liposome, and is connected with CA9.More specifically, with CA9 connected mode for CA9 antibody is connected to liposome Polyethylene Glycol chain end.
In further specific embodiment, the membrane material of Venenum Bufonis target liposomes is selected from following material: the phospholipid of a, native soy lecithin or synthesis; B, cholesterol; The compositions of c, polyethylene glycol-phosphorus fat complexes and d, CAC A9 antibody-polyethylene glycol-phosphorus fat complexes.In described material c, in polyethylene glycol-phosphorus fat complexes, the molecular weight of Polyethylene Glycol is 1000-5000g/mol; In described material d, in CAC A9 antibody-polyethylene glycol-phosphorus fat complexes, the molecular weight of Polyethylene Glycol is 1000-8000g/mol.
In further specific embodiment, the mol ratio of liposome membrane material is a: b=4:1-1:1, a:c=1000:1-1000:100, a:d=1000:1-1000:100.
In further specific embodiment, the mass ratio of Venenum Bufonis and described lipid composition is 0.1% to 50%.
In further specific embodiment, liposomal particle size scope is 20-200nm.
In further specific embodiment, the phospholipid in described polyethylene glycol-phosphorus fat complexes or CAC A9 antibody-polyethylene glycol-phosphorus fat complexes is selected from DSPE, DPPE, DOPE, hydrogenated soya phosphatide acyl ethanolamine, hydrogenation egg phosphatide acyl ethanolamine, soybean phospholipid phosphatidyl ethanolamine or egg phosphatide acyl ethanolamine.
(2) preparation method of Venenum Bufonis target liposomes
The preparation method of Venenum Bufonis target liposomes of the present invention, carry out in accordance with the following steps:
1) Chansu Liposome is prepared
Use extruder to extrude through the several circulation of polycarbonate filter, control particle diameter within 20-200nm.
2) Venenum Bufonis target liposomes is prepared
Add Chansu Liposome, 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride, N-hydroxy thiosuccinimide, CAC A9 antibody, under being placed in 4 DEG C of conditions, react 4-20 hour;
3) purification
Use bag filter is dialysed, and removes non-encapsulated Venenum Bufonis, Chansu Liposome, the CAC A9 antibody do not connected, EDCHCl, S-NHS;
4) dry
To the Venenum Bufonis target liposomes lyophilization obtained of dialysing, obtain powdered samples, so that preserve.
In further specific embodiment, in described step 1, the final concentration of Chansu Liposome controls at 5-20mg/ml.
In further specific embodiment, in step 2, Chansu Liposome, 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride, N-hydroxy thiosuccinimide, CAC A9 antibody ratios are (10-100): (100-1000): (100-1000): (1-10).
In further specific embodiment, in described step 2, the response time is 8-12 hour.
Venenum Bufonis target liposomes of the present invention can be applied to prepares Therapeutic cancer medicine, preferably prepares cancer drug and comprises colon cancer drug, bladder cancer medicine, renal carcinoma medicine, cervical cancer medicine, cerebral glioma medicine, gastric cancer medicament.
3, beneficial effect of the present invention.
(1) the present invention is by the synergism by Chansu Liposome and CA9, can reduce the drug toxicity of Venenum Bufonis, decrease the injury to human normal cell.
(2) the present invention is by the combination of Chansu Liposome and CA9, and its therapeutic effect is also far above the therapeutic effect of Chansu Liposome of the prior art, and tumour inhibiting rate is up to more than 60%.
(3) the present invention is by by the preparation technology of Chansu Liposome and CA9 and synergism, can arrive cancer focal zone targetedly, and does not provide CA9 in prior art and how specifically to prepare and to be applied to the method for Therapeutic cancer medicine.
(4) Venenum Bufonis targeting lipids preparation process of the present invention is easy, CAC A9 antibody and Chansu Liposome coupling efficiency reach 68%-70%, and envelop rate can reach 80-85%, particle diameter 20-200nm, stability is better, and Venenum Bufonis target liposomes 16 days slips in PBS solution are only 3.4%.
Accompanying drawing explanation
Fig. 1 Venenum Bufonis target liposomes prepares schematic diagram
The time dependent permeability of Fig. 2 Venenum Bufonis target liposomes
Fig. 3 Venenum Bufonis target liposomes is on the impact of colon cancer HCT116 cell survival rate
Fig. 4 Venenum Bufonis target liposomes is on the impact of cervical cancer hela cell survival rate
Fig. 5 Venenum Bufonis target liposomes is on the impact of renal carcinoma 796-P cell survival rate.
Detailed description of the invention
In order to enable the auditor of Patent Office especially the public clearly understand technical spirit of the present invention and beneficial effect, applicant will elaborate below by way of example, but be not all the restriction to the present invention program to the description of embodiment, any conceive according to the present invention done be only pro forma but not substantial equivalent transformation and all should be considered as technical scheme category of the present invention.
the preparation method of embodiment 1 Venenum Bufonis target liposomes
1, the preparation of Chansu Liposome is wrapped up
1) distearoyl phosphatidylcholine (DSPC) 60mg is got, cholesterol (Chol) 10mg, distearyl acid PHOSPHATIDYL ETHANOLAMINE-Macrogol 2000 (DSPE-PEG2000) 9mg, distearyl acid PHOSPHATIDYL ETHANOLAMINE-Macrogol 2000-carboxyl commissure thing (DSPE-PEG2000-COOH) 1.5mg is dissolved in (solution A) in 15ml chloroform; Venenum Bufonis is dissolved in (B solution) in methanol, and solution A mixes according to 1:1 ratio with B solution, and adopt film dispersion method, 60 DEG C, 40r/min, boils off chloroform, methanol.
2) by this lipid film in the isotonic salt buffer of the Tris-of 10mM (Tris of 10mM, 137mM sodium chloride, 25 DEG C, pH7.4) aquation.The final concentration of lipid is controlled at 5-20mg/ml.
3) then 60min is kept to solidify to make described liposome structure 80 DEG C (exceeding the transition temperature of all lipids) in described mixture.During curing, respectively at time point that is that start, middle and end by its with vortex stirring 3 times, and continuous stirring 5min each time.
4) finally 10ml extruder is used to extrude with several circulation, until particle diameter is within 20-200nm through the Whatman100nm polycarbonate filter that diameter is 25mm the MLV obtained.
, targeting lipids preparation
As shown in Figure 1, the preparation flow of Venenum Bufonis target liposomes:
1) 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (EDC.HCL) is got and N-hydroxy thiosuccinimide (S-NHS) adds in the liposome PBS buffer of the parcel Venenum Bufonis of above-mentioned preparation (pH7.4), after stirring at room temperature 30min, add CAC A9 antibody, under being placed in 4 DEG C of conditions, react 12h.
2) purification: be that the PBS of 10mM is as dialysis medium by pH7.4, concentration, bag filter dialysis 1h, change dialysis medium, repeat aforesaid operations 2 times, remove non-encapsulated Venenum Bufonis, unreacted CAC A9 antibody, EDCHCl, S-NHS and conventional liposome.
Dry: to the Venenum Bufonis target liposomes lyophilization obtained of dialysing, to obtain powdered samples, so that preserve.
the sign of embodiment 2 Venenum Bufonis target liposomes
1, the mensuration of envelop rate
By 150 μ L freshly prepd Venenum Bufonis target liposomes AvantiJ-E centrifuge (JA-20,17400*g, 6 DEG C, 20min) by can molecular cut off to be that 10KD considers the MicroconY-10 centrifugal filter device (Millipore) of part centrifugal.By the Venenum Bufonis concentration in spectrophotometry centrifugal solution under 296nm.This concentration represents the Venenum Bufonis concentration (Venenum Bufonis of non-encapsulated) in the continuous phase of described liposome.Spectrophotography is also used as to measure the Venenum Bufonis total concentration in the decentralized photo of liposome and continuous phase.Dehydrated alcohol is added in liposome turbid liquor to break described liposome and being dissolved completely in solution by the Venenum Bufonis of encapsulating completely.Under the wavelength of 296nm, absorbance is measured by the UV-VISVarianCary50 sub-ray spectrometer being equipped with constant temperature quartz chamber.By following formula computational envelope rate (EE):
EE (%)=(C
always-C
free)/C always × 100
Wherein C
freethe non-encapsulated Venenum Bufonis concentration in the continuous phase of liposomal dispersion/solution, the Venenum Bufonis total concentration of C always in liposomal dispersion/solution.Through calculating, envelop rate can reach 80-85%.
, Chansu Liposome particle diameter
Emulsion droplet granulometry at present about liposome often adopts method to have microscopic method, dynamic light scattering method, Coulter counting method etc.In this experiment, the granulometry of Venenum Bufonis target liposomes adopts optical microscope and photon correlation spectroscopy (Photon-CorrelationSpectroscopy, PCS), also dynamic light scattering principle (Dynamiclightscatter, DLS, NicompTMPSS380) is claimed.
The operational approach using NicompTMPSS380 to carry out granulometry is: the water for injection dilution about 5000 times sample being used 0.22um microporous filter membrane, put into sample cell immediately, light intensity (Intensity) is adjusted to about 300, and light source is HeNe laser (λ
0=633nm), temperature in operating parameter is set as room temperature, starts to measure, keep measuring and become to stopping during straight line measuring to Timehistory curve, preserve data.Particle diameter 20 ~ 200nm.
, Venenum Bufonis target liposomes medicine stability
In order to measure the stability of liposome Venenum Bufonis, with 25mlPBS(pH7.4) respectively at the Venenum Bufonis target liposomes (molecular cut off 8000) of 37 DEG C of dialysis 500ul.Dialysis solution (500ul) respectively shifted once at 0,0.5,1,2,4,8,16 day, was carried out the amount of Venenum Bufonis in detection by quantitative dialysis solution by HPLC.Computing permeability formula: permeability=product leaks into medicine × 100% that the dose/product in medium is encapsulated before storage after stored for some time.
As shown in Figure 2, Venenum Bufonis target liposomes 16 days slips in PBS solution are only 3.4%, and what show that Venenum Bufonis can be stable is wrapped in liposome, not easily reveals.But consider the stability of CAC A9, Venenum Bufonis target liposomes medicament freeze-drying is become powder, is easy to long-term preservation.
, CA9 antibody and Chansu Liposome coupling efficiency mensuration
Liposome can disturb albumen direct-detection strongly, and such as Lowry or BCA detects.Therefore, the combination of conbined usage post separation, Protein Detection, the quantitative CA9 antibody of ELISA and surface of liposome.Freshly prepd Venenum Bufonis target liposomes joins SepharoseCL-4B, and the component of every 10 is collected together separately, uses BCA protein reagent box analyzing proteins content.The component comprising liposome uses this kind of detection method there is interference, therefore there is false positive results.Therefore this part of ELISA detects, to confirm the existence of CA9 antibody.
In order to the combination degree of quantitative CA9 antibody and liposome, we adopt protein detection kit to formulate the standard curve of antibody by the series concentration of antibody in PBS.The antibody amount in conjunction with liposome eluted from pillar is quantitative by standard curve.By being covalently bound to antibody amount on liposome by deducting unconjugated antibody preparing in Venenum Bufonis target liposomes process total antibody amount of adding.Experimental result shows, CAC A9 antibody and Chansu Liposome coupling efficiency are 68%-70%.
embodiment 3, Venenum Bufonis target liposomes are on the impact (MTT colorimetry) of tumor cell viability
After the HCT116 of trophophase of taking the logarithm is cells trypsinised, culture fluid dilutes, and makes 3 × 10
4/ ml cell suspension, is inoculated in 96 well culture plates, every hole 100 μ l.Experimental group, negative control group and blank group are established in experiment.Cultivate 24 hours after inoculation, add each group of sample 25 μ L respectively, each repetition 5 hole, after 37 DEG C of 5%CO2 cultivate 0h, 4h, 8h, 16h, 24h respectively, every hole adds MTT5mg/ml20 μ l, continues to hatch 4h in 37 DEG C of incubators.96 orifice plates are discarded upper liquid, and every hole adds dimethyl sulfoxine (DMSO) 120 μ L, and agitator low speed jolting 10min, makes abundant dissolving, microplate reader is 492nm measures absorbance (OD) value with determined wavelength.Cell survival rate %=[(cancer therapy drug group OD value-blank group OD value)/(negative control group OD value-blank group OD value)] × 100%.
Cervical cancer cell hela, kidney cancer cell 769-P cultivate according to the method described above, detect cell survival rate.
Colon cancer HCT116 cell, cervical cancer hela cell, renal carcinoma 796-P cell survival rate all can be suppressed by the known Venenum Bufonis solution of Fig. 3, Fig. 4, Fig. 5, Chansu Liposome, Venenum Bufonis target liposomes.Wherein Venenum Bufonis solution inhibition is obviously better than Chansu Liposome and Venenum Bufonis target liposomes, reason is that Venenum Bufonis solution directly can contact tumor cell, and the Venenum Bufonis wrapped up in Chansu Liposome, Venenum Bufonis target liposomes needs slow releasing, and release is complete not, so Venenum Bufonis solution can more rapidly (4h), more efficient in killing tumor cell process.But Venenum Bufonis solution is also larger to the toxic action of the cell of other health, should not directly apply also is the difficult problem run in prior art.
Can see that the Chansu Liposome prepared by embodiment 1 and Venenum Bufonis target liposomes all have lethal effect to colon cancer HCT116 cell, cervical cancer hela cell, renal carcinoma 796-P cell by the present embodiment.Wherein Venenum Bufonis target liposomes does not affect the release of Venenum Bufonis medicine because connecting CA9 antibody, and its targeting effect is by pharmacodynamics checking below.
embodiment 4 Venenum Bufonis target liposomes is to tumor growth pharmacodynamic evaluation
The lotus tumor nude mouse 30 having been inoculated colon cancer HCT116 cell is divided into 5 groups at random, often organizes 6.Arrange normal saline group, Venenum Bufonis group, Chansu Liposome group, Venenum Bufonis target liposomes small dose group, Venenum Bufonis target liposomes high dose group, wherein normal saline group is negative control group.Each group of mice is normally raised, administration from 24h after inoculation, once a day, totally 7 times, each tail vein injection said medicine 0.2ml.Claim Mouse Weight after 7 days, by tumor sacrifice, dissect, get tumor, and claim tumor weight.Tumour inhibiting rate=[(negative control group average tumor weight-treatment group average tumor weight) average tumor weight of/negative control group] * 100%, the data of acquisition represent with (± SDg), and do t inspection.
Cervical cancer cell hela, kidney cancer cell 769-P make lotus tumor nude mouse model, and then administration according to the method described above, detects tumour inhibiting rate after 7 days respectively.
Table 1 Venenum Bufonis and target liposomes thereof are to mouse junction cancer inhibitory action (n=6)
Illustrate:
Compared with normal saline group, all the other are respectively organized tumor weight average and have significant difference (P<0.01).
﹡, compared with Venenum Bufonis solution group, has significant difference (P<0.01).
#, compared with Chansu Liposome group, has significant difference (P<0.01).
Table 2 Venenum Bufonis and target liposomes thereof are to mouse cervical cancer inhibitory action (n=6)
Illustrate:
Compared with normal saline group, all the other are respectively organized tumor weight average and have significant difference (P<0.01).
﹡, compared with Venenum Bufonis solution group, has significant difference (P<0.01).
#, compared with Chansu Liposome group, has significant difference (P<0.01).
Table 3 Venenum Bufonis and target liposomes thereof are to mice renal carcinoma inhibitory action (n=6)
Illustrate:
Compared with normal saline group, all the other are respectively organized tumor weight average and have significant difference (P<0.01).
﹡, compared with Venenum Bufonis solution group, has significant difference (P<0.01).
#, compared with Chansu Liposome group, has significant difference (P<0.01).
From table 1, table 2, table 3, Venenum Bufonis can suppress the growth of mouse junction cancer, cervical cancer, renal carcinoma, and wherein Venenum Bufonis target liposomes effect is particularly remarkable.Reason is that Venenum Bufonis target liposomes is connected to CAC A9 antibody, can combine, make the Venenum Bufonis orientation of wrapping up in liposome in tumor cell with the tumor cell targeting of high expressed CAC A9.In experimentation, the mice part of injection Venenum Bufonis solution there will be diarrhoea, inappetence, convulsions symptom, shows some side effect of Venenum Bufonis, and injection Venenum Bufonis target liposomes group is but without this side effect.
Claims (14)
1. a Venenum Bufonis target liposomes, is characterized in that: comprise CAC A9 antibody, Venenum Bufonis and liposome, Venenum Bufonis is wrapped in liposome, and liposome is connected with CA9.
2. Venenum Bufonis target liposomes according to claim 1, is characterized in that: with CA9 connected mode for CA9 antibody is connected to liposome Polyethylene Glycol chain end.
3. Venenum Bufonis target liposomes according to claim 1 and 2, is characterized in that the membrane material of its liposome is selected from following material: the phospholipid of a, native soy lecithin or synthesis; B, cholesterol; The compositions of c, polyethylene glycol-phosphorus fat complexes and d, CAC A9 antibody-polyethylene glycol-phosphorus fat complexes.
4. Venenum Bufonis target liposomes according to claim 3, is characterized in that: in described material c, in polyethylene glycol-phosphorus fat complexes, the molecular weight of Polyethylene Glycol is 1000-5000g/mol; In described material d, in CAC A9 antibody-polyethylene glycol-phosphorus fat complexes, the molecular weight of Polyethylene Glycol is 1000-8000g/mol.
5. Venenum Bufonis target liposomes according to claim 3, is characterized in that: the mol ratio of liposome membrane material is a: b=4:1-1:1, a:c=1000:1-1000:100, a:d=1000:1-1000:100.
6. Venenum Bufonis target liposomes according to claim 1, is characterized in that: the mass ratio of Venenum Bufonis and described lipid composition is 0.1% to 50%.
7. Venenum Bufonis target liposomes according to claim 1, is characterized in that: liposomal particle size scope is 20-200nm.
8. Venenum Bufonis target liposomes according to claim 3, is characterized in that: the phospholipid in described polyethylene glycol-phosphorus fat complexes or CAC A9 antibody-polyethylene glycol-phosphorus fat complexes is selected from DSPE, DPPE, DOPE, hydrogenated soya phosphatide acyl ethanolamine, hydrogenation egg phosphatide acyl ethanolamine, soybean phospholipid phosphatidyl ethanolamine or egg phosphatide acyl ethanolamine.
9. a preparation method for Venenum Bufonis target liposomes, is characterized in that carrying out in accordance with the following steps:
1) Chansu Liposome is prepared
Use extruder to extrude through the several circulation of polycarbonate filter, control particle diameter within 20-200nm;
2) Venenum Bufonis target liposomes is prepared
Add Chansu Liposome, 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride, N-hydroxy thiosuccinimide, CAC A9 antibody, under being placed in 4 DEG C of conditions, react 4-20 hour;
3) purification
Use bag filter is dialysed, and removes non-encapsulated Venenum Bufonis, Chansu Liposome, the CAC A9 antibody do not connected, EDCHCl, S-NHS;
4) dry
To the Venenum Bufonis target liposomes lyophilization obtained of dialysing, obtain powdered samples, so that preserve.
10. the preparation method of Venenum Bufonis target liposomes according to claim 9, is characterized in that: in described step 1, the final concentration of Chansu Liposome controls at 5-20mg/ml.
The preparation method of 11. Venenum Bufonis target liposomes according to claim 9, is characterized in that: in step 2, Chansu Liposome, 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride, N-hydroxy thiosuccinimide, CAC A9 antibody ratios are (10-100): (100-1000): (100-1000): (1-10).
The preparation method of 12. Venenum Bufonis target liposomes according to claim 9, is characterized in that: in described step 2, the response time is 8-12 hour.
13. 1 kinds of Venenum Bufonis target liposomes are for the preparation of the application of Therapeutic cancer medicine.
14. Venenum Bufonis target liposomes according to claim 14, for the preparation of the application of cancer treatment drugs, is characterized in that: cancer drug comprises colon cancer drug, bladder cancer medicine, renal carcinoma medicine, cervical cancer medicine, cerebral glioma medicine, gastric cancer medicament.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106420618A (en) * | 2016-09-23 | 2017-02-22 | 苏州卫生职业技术学院 | Preparation method for Anti-CA IX modified norcantharidin micelle nanoparticle |
| CN111150856A (en) * | 2019-11-26 | 2020-05-15 | 宁波市鄞州区第二医院医共体 | Ultrasonic molecular probe and preparation method thereof |
| CN116196279A (en) * | 2023-04-27 | 2023-06-02 | 上海中医药大学 | Cholesterol-free liposome of toad venom extract and its preparation method and application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102429868A (en) * | 2011-12-09 | 2012-05-02 | 南开大学 | Liposome pharmaceutical composition with tumor targeting, in vivo tracing and treating effects and preparation method thereof |
| CN103083239A (en) * | 2012-12-26 | 2013-05-08 | 中国人民解放军第四军医大学 | Bufalin lipidosome, preparation method and application thereof |
-
2014
- 2014-10-15 CN CN201410544846.1A patent/CN105561304A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102429868A (en) * | 2011-12-09 | 2012-05-02 | 南开大学 | Liposome pharmaceutical composition with tumor targeting, in vivo tracing and treating effects and preparation method thereof |
| CN103083239A (en) * | 2012-12-26 | 2013-05-08 | 中国人民解放军第四军医大学 | Bufalin lipidosome, preparation method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| BLENDA CHI KWAN WONG等: "Carbonic anhydrase IX-directed immunoliposomes for targeted drug delivery to human lung cancer cells in vitro", 《DRUG DESIGN, DEVELOPMENT AND THERAPY》 * |
| SHINKAI M等: "Targeting hyperthermia for renal cell carcinoma using human MN antigen-specific magnetoliposomes", 《JPN J CANCER RES》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106420618A (en) * | 2016-09-23 | 2017-02-22 | 苏州卫生职业技术学院 | Preparation method for Anti-CA IX modified norcantharidin micelle nanoparticle |
| CN106420618B (en) * | 2016-09-23 | 2019-05-03 | 苏州卫生职业技术学院 | A kind of preparation method of norcantharidin micellar nanoparticles modified by Anti-CA IX |
| CN111150856A (en) * | 2019-11-26 | 2020-05-15 | 宁波市鄞州区第二医院医共体 | Ultrasonic molecular probe and preparation method thereof |
| CN116196279A (en) * | 2023-04-27 | 2023-06-02 | 上海中医药大学 | Cholesterol-free liposome of toad venom extract and its preparation method and application |
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