CN103055302A - Ultrasonic biological effect mediated recombinant human endostatin controlled release preparation - Google Patents
Ultrasonic biological effect mediated recombinant human endostatin controlled release preparation Download PDFInfo
- Publication number
- CN103055302A CN103055302A CN2013100062751A CN201310006275A CN103055302A CN 103055302 A CN103055302 A CN 103055302A CN 2013100062751 A CN2013100062751 A CN 2013100062751A CN 201310006275 A CN201310006275 A CN 201310006275A CN 103055302 A CN103055302 A CN 103055302A
- Authority
- CN
- China
- Prior art keywords
- recombinant human
- drug
- preparation
- biological effect
- release
- 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
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses an ultrasonic biological effect mediated recombinant human endostatin controlled release preparation. A preparation method of the biological effect mediated recombinant human endostatin controlled release preparation comprises the following steps: taking 2mg of recombinant human endostatin, adding 1ml of a medical fibrin glue main glue solution with the fibrinogen concentration of 50mg/ml, uniformly mixing, and mixing with 1ml of a catalyst solution containing 400IU of thrombin to form a drug loaded biogel; and placing the drug loaded biogel in 10ml of a phosphate buffer solution having a concentration of 0.01mol/L, having a pH value of 7.2 and containing 0.2mg/ml of NaN3, and shaking on a 37DEG C constant temperature shaking table at a speed of 80times/min for in-vitro release, wherein the release amount is 21.5% 2h later, and the accumulated drug release amount reaches 89.7% 5 days later. Ultrasonic irradiation obviously promotes the drug release speed of the biogel, so the ultrasonic biological effect mediated recombinant human endostatin controlled release preparation has the drug release control effect, and can be applied to the preparation of biological effect mediated tumor blood vessel generation inhibition drugs; and the tumor inhibition rate of the ultrasonic biological effect mediated recombinant human endostatin controlled release preparation reaches 53.2%.
Description
Technical field
The present invention relates to a kind of biogum preparation of load-carrying group human Endostatin and the application in the inhibition tumor-blood-vessel growth medicine under the biological effect mediation that preparation ultrasonic in combination microbubble contrast agent produces thereof.
Background technology
Vascellum esoderma inhibin is a kind of pharmaceutical grade protein with inhibition angiogenesis of discovered in recent years, and in new vessels endotheliocyte and cell growth inhibiting and migration, cell death inducing is brought into play blood vessel formation against function by specific effect for it.In addition, but vascellum esoderma inhibin also inhibition tumor cell vascular surface endothelial growth factor expression and hydrolase of proteolysis, many target spots performance blood vessel formation against function.Recombinant human vascular endothelial inhibin (recombinant human endostatin, rh-ES) has higher purity and stability than vascellum esoderma inhibin, and anti-angiogenesis activity is stronger, has been applied to clinical antitumor.But because rh-ES lacks specificity to body normal blood vessels and tumor neogenetic blood vessels effect, simple intravenous administration, the tumor by local drug level is low, and antitumor action is limited, and has untoward reaction.In addition, because the rh-ES half-life is short, need intravenously administrable every day, patient compliance is poor.
The Multiple components such as the Fibrinogen that biological fibrin glue is extracted by animal blood, thrombin, calcium ion form, the final stage of simulation human body blood coagulation mechanism, formed protein gel has the effects such as hemostasis, bonding, wound closure after mixing, and has been applied to clinical.Research finds that also the similar drug storehouse storage of the fibrous reticular structure of biological fibrin glue is a kind of good medicine control slow-released carrier.Biological fibrin glue is in vivo along with the activation of fibrinolysin, gradually solution absorption.In addition, adopt biological fibrin glue preparation control slow releasing pharmaceutical, only need to get final product injecting behind medicine and the colloid solution mixing, need not with an organic solvent, the pharmaceutical grade protein activity is had no adverse effects.Adopting biological fibrin glue is that carrier prepares the biological activity that the recombinant human vascular endothelial inhibin controlled slow-release preparation can keep medicine, and body is had no side effect.
Injection for curing can obviously improve the tumor by local drug level in the control slow releasing pharmaceutical tumor, and then improves antitumous effect, and obviously reduces administration number of times, and is easy to operate.But Recent study shows, after the injection, is absorbed after some drugs discharges and enters blood circulation in the control slow releasing pharmaceutical tumor.Therefore, making to greatest extent medicine be retained in tumor by local is to improve antitumor curative effect, alleviates the key of toxic and side effects.The main target spot of vascellum esoderma inhibin effect is positioned at vascular endothelial cell, and injection rh-ES biogum in the tumor only has when drug release and enters blood circulation competence exertion blood vessel formation against function.But enter sanguimotor medicine and will arrive whole body with blood flow very soon, reality can be retained in tumor by local, and vasoactive medication amount is few, and the antineoplastic vascular nucleus formation is limited.
Summary of the invention
The purpose of this invention is to provide a kind of biogum preparation of load-carrying group human Endostatin and the application in the inhibition tumor-blood-vessel growth medicine under the biological effect mediation that preparation ultrasonic in combination microbubble contrast agent produces thereof.
The object of the present invention is achieved like this, the recombinant human vascular endothelial inhibin controlled release preparation of described a kind of biological effect of ultrasound mediation, be prepared from by following method, get the 2mg recombinant human vascular endothelial inhibin, adding 1ml fibrinogen concentration is the main gel solution of the Fibrin Glue of 50mg/ml, then mixing mixes with the catalyst solution that 1ml contains the 400IU thrombin, forms drug-loaded biological glue; It is that 0.01mol/L, pH are 7.2 and contain 0.2mg/mlNaN that drug-loaded biological glue is placed 10ml concentration
3Phosphate buffer in, in 37 ℃ of constant-temperature tables, shake with 80 times/component velocity, carry out release in vitro, ELISA measures drug-loaded biological glue and discharges medication amount, and drug-loaded biological glue is at the external recombinant human vascular endothelial inhibin that more steadily discharges, 2h discharges 21.5%, 5 day cumulative release medication amount and reaches 89.7%; Ultrasonic irradiation obviously promotes biogum to discharge the speed of medicine, has the effect of control drug release.
Recombinant human vascular endothelial inhibin biogum preparation of the present invention suppresses the application in the tumor-blood-vessel growth medicine under the biological effect mediation that preparation ultrasonic in combination microbubble contrast agent produces.
Ultrasonic irradiation biological tissue can produce heat effect, mechanical effect, cavitation effect etc., and when these physical effects were accumulated to a certain degree in vivo, the rising of local organization temperature, blood vessel wall and permeability of cell membrane increased, i.e. biological effect of ultrasound.Simultaneously, the physical effect that ultrasonic irradiation produces can be controlled the rate of release of slow releasing preparation, especially at the drug releasing rate of body gel, reaches the effect of control drug release of fixed place and time.Studies show that in a large number ultrasonic irradiation breaks up medicine-carrying microvesicle can obviously strengthen biological effect of ultrasound, the blood capillary of diameter≤7 μ m is broken, the broadening of endotheliocyte gap, permeability of cell membrane increases, the local organization microcirculation disturbance.
Advantage of the present invention is: the present invention adopts biological fibrin glue bag load-carrying group human Endostatin to prepare drug-loaded biological glue preparation, have no side effect, after injecting in the tumor, control drug releasing rate by the biological effect that ultrasonic irradiation produces, make slow releasing preparation reach the effect of controlled release, adjustable point discharges medicine at regular time and quantity.Make tumor by local vascular endothelial cell gap enlargement by the ultrasonic irradiation microbubble contrast agent, controlled the medicine that discharges and to permeate in blood capillary in a large number.The powerful biological effect that the ultrasonic irradiation microbubble contrast agent produces makes the tumor by local blood capillary inaccessible, circulatory disturbance, and being penetrated into endovascular medicine can be retained in the tumor by local blood vessel for a long time, directly acts on vascular endothelial cell.In addition, the tumor by local blood circulatory disorder, fibrinoclase enters the tumor by local amount and obviously reduces in the blood, the degradation speed of biological fibrin glue slows down, and can reach more lasting control slow release effect, inhibitory rate to 53.2%, be about 5 times of rh-ES biogum intratumor injection group, obvious effect is arranged.
Description of drawings
Fig. 1 is the release in vitro figure of drug-loaded biological glue of the present invention.
The specific embodiment
The present invention is described in detail below in conjunction with drawings and Examples:
Embodiment one
Get 2mg rh-ES, add 1ml Fibrin Glue (Guangzhou Beixiu Biological Technology Co., Ltd produces) main gel solution (fibrinogen concentration is 50mg/ml), mixing mixes with 1ml catalyst solution (containing thrombin 400IU), forms drug-loaded biological glue.Drug-loaded biological glue is placed the 10ml phosphate buffer, and (0.01mol/L, pH7.2 contain 0.2mg/ml NaN
3), in 37 ℃ of constant-temperature tables, 80 times/minutes, release in vitro is carried out in concussion, and ELISA measures and discharges medication amount.Drug-loaded biological glue is at the external rh-ES that more steadily discharges, and 2h discharges 21.5%, 5 day cumulative release medication amount and reaches 89.7%; See Fig. 1.
Above-mentioned Fibrin Glue: produce and provide by Guangzhou Beixiu Biological Technology Co., Ltd, be the commercially available prod.Every cover " Fibrin Glue " comprises main gel (the present invention is mixed with including fiber proteinogen concentration when embodiment one experiment be 50mg/ml) and the catalyst (the present invention is mixed with and contains thrombin 400IU in the 1ml catalyst solution) each bottle (being powdery) of Fibrin Glue when embodiment one experiment, and with each 1.5 milliliters of corresponding lytic agents.
Embodiment two
Adopt the drug-loaded biological glue of ultrasonic irradiation release in vitro, the sound intensity is 1.0 W/cm
2, behind the irradiation 3min, 10min, 30min respectively take a sample 1 time, measure release amount of medicine.Found that ultrasonic irradiation obviously promotes biogum to discharge the speed of medicine, has the effect of control drug release.
Table 1 ultrasonic irradiation is on drug-loaded biological glue drug release rate impact (%)
| Group | 10min | 30min |
| Matched group | 5.2±1.6 | 10.1±3.3 |
| The ultrasonic irradiation group | 15.7±6.8 ** | 19.5±7.5 ** |
Annotate: compare with matched group,
* P＜0.01.
Embodiment three
In vitro culture human breast cancer cell MDA-MB-231, after a large amount of amplifications, it is subcutaneous to be inoculated into nude mouse right fore root, sets up lotus human breast carcinoma nude mice by subcutaneous transplanted tumor model.Choose 15 of tumor bearing nude mices, tumor maximum diameter 1.2cm～1.5cm, be divided at random 3 groups, every group 5, be respectively (A group) rh-ES biogum intratumor injection group, (B group) Erh-ES biogum intratumor injection+ultrasonic irradiation group, (C group) rh-ES biogum intratumor injection+microbubble contrast agent+ultrasonic irradiation group.A group intratumor injection drug-loaded biological glue, rh-ES dosage is 10mg/kg, the biogum volume injected is 0.5ml; B organizes behind intratumor injection 10min, 1.0 W/cm
2Ultrasonic irradiation tumor tissues 3min; Behind the C group nude mice intratumor injection administration 10min, behind the tail vein injection acoustic contrast agent SonoVue, carry out immediately tumor ultrasonic irradiation 3min.Get blood examination behind the 30min and survey serum rh-ES concentration.The result, ultrasonic irradiation group nude mice blood drug level is (7.47 ± 1.52) ng/mL, (3.11 ± 0.98) ng/mL apparently higher than simple intratumor injection group, and ultrasonic irradiation combining ultrasonic contrast agent group blood drug level is starkly lower than simple intratumor injection group, only be (1.32 ± 0.36) ng/mL, each organize comparing difference have statistical significance (
P＜0.01).
Embodiment four
Choose 25 of tumor bearing nude mices, the about 1.5cm of tumor maximum diameter is divided into 5 groups at random, 5 every group.A: model group; The B:rh-ES group, intratumor injection rh-ES injection; C:rh-ES biogum group, intratumor injection rh-ES biogum; D:rh-ES biogum+ultrasonic irradiation group, row tumor ultrasonic irradiation behind the intratumor injection rh-ES biogum; E:rh-ES biogum+microbubble contrast agent+ultrasonic irradiation group, intratumor injection rh-ES biogum, SonoVue annotates in tail vein group behind the 10min, carries out immediately the tumor ultrasonic irradiation.
More than process per 5 days 1 courses for the treatment of, repeat 4 times, rh-ES dosage is 10mg/kg, and volume injected is 0.5ml.The 1st day of each course for the treatment of, each organizes nude mice intratumor injection medicine, and D group nude mice carries out ultrasonic irradiation, E group nude mice injection acoustic contrast agent and ultrasonic irradiation, the 3rd day, D group nude mice was repeated ultrasonic irradiation, E group nude mice duplicate injection acoustic contrast agent and ultrasonic irradiation.
Embodiment five
Rear the 20th day for the treatment of, each is organized nude mice tail vein injection acoustic contrast agent SonoVue and carries out ultrasonic contrast observation tumor blood circulation perfusion situation.Enable Syngo US Workplace software and analyze, the beginning of respectively organizing the tumor ultrasonic contrast increases time (AT), peak time (TTP), peak value underlying strength poor (PBD).As a result, the PBD of E group tumor be starkly lower than all the other each groups (
P＜0.01).
Table 2 is respectively organized tumor ultrasonic contrast parameter relatively
| Group | AT | TTP | PBD |
| A | 1.95±0.42 | 3.97±0.86 | 22.14±5.83 |
| B | 1.91±0.37 | 4.08±0.93 | 19.62±4.48 |
| C | 2.01±0.45 | 4.01±0.97 | 18.37±4.28 |
| D | 1.87±0.39 | 3.85±0.91 | 17.56±4.36 * |
| E | 1.90±0.33 | 3.92±0.89 | 8.37±2.97 *# |
Annotate: compare with the A group,
* P＜0.05; Compare with the D group,
# P＜0.01.
Embodiment six
Behind the nude mice ultrasonic contrast, etherization is put to death, and peels off tumor and claims quality, calculates inhibition rate of tumor growth according to formula.Suppression ratio (%)=(the average tumor quality of the average tumor quality/model group of 1-treatment group) * 100%.As a result, E group tumor growth suppresses the most obvious, inhibitory rate 53.2%.
Tumor quality respectively organized by table 3 and tumour inhibiting rate compares
| Group | Tumor quality (g) | Tumour inhibiting rate (%) |
| A | 3.22±0.97 | - |
| B | 2.85±0.61 | 11.5 |
| C | 2.53±0.58 | 21.4 |
| D | 2.21±0.49 | 31.4 |
| E | 1.51±0.37 | 53.2 |
Claims (2)
1. the recombinant human vascular endothelial inhibin controlled release preparation of biological effect of ultrasound mediation, be prepared from by following method, get the 2mg recombinant human vascular endothelial inhibin, adding 1ml fibrinogen concentration is the main gel solution of the Fibrin Glue of 50mg/ml, mixing, then mix with the catalyst solution that 1ml contains the 400IU thrombin, form drug-loaded biological glue; It is that 0.01mol/L, pH are 7.2 and contain 0.2mg/mlNaN that drug-loaded biological glue is placed 10ml concentration
3Phosphate buffer in, in 37 ℃ of constant-temperature tables, shake with 80 times/component velocity, carry out release in vitro, ELISA measures drug-loaded biological glue and discharges medication amount, and drug-loaded biological glue is at the external recombinant human vascular endothelial inhibin that more steadily discharges, 2h discharges 21.5%, 5 day cumulative release medication amount and reaches 89.7%; Ultrasonic irradiation obviously promotes biogum to discharge the speed of medicine, has the effect of control drug release.
2. recombinant human vascular endothelial inhibin biogum preparation claimed in claim 1 suppresses the application in the tumor-blood-vessel growth medicine under the biological effect mediation that preparation ultrasonic in combination microbubble contrast agent produces.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310006275.1A CN103055302B (en) | 2013-01-08 | 2013-01-08 | Ultrasonic biological effect mediated recombinant human endostatin controlled release preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310006275.1A CN103055302B (en) | 2013-01-08 | 2013-01-08 | Ultrasonic biological effect mediated recombinant human endostatin controlled release preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103055302A true CN103055302A (en) | 2013-04-24 |
| CN103055302B CN103055302B (en) | 2014-05-14 |
Family
ID=48098395
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310006275.1A Expired - Fee Related CN103055302B (en) | 2013-01-08 | 2013-01-08 | Ultrasonic biological effect mediated recombinant human endostatin controlled release preparation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103055302B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110917332A (en) * | 2019-12-12 | 2020-03-27 | 南通大禹生物技术有限公司 | Endodu in-vivo controlled-release medicine and preparation method and application thereof |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1544098A (en) * | 2003-11-12 | 2004-11-10 | 邹金生 | Highly effective NGF adhesive and its preparation and using method |
| CN1946352A (en) * | 2004-04-09 | 2007-04-11 | 狄科特康坦特公司 | Methods and articles for the delivery of medicaments to the eye for the treatment of posterior segment diseases |
| CN101020061A (en) * | 2007-03-06 | 2007-08-22 | 西安交通大学 | Prepn process of ultrasonically controlled releasing micro gel containing medicine |
| CN101536984A (en) * | 2008-03-20 | 2009-09-23 | 江苏先声药物研究有限公司 | Injection-use recombinant human Endostatin porous sustained-release microsphere and preparation method thereof |
| CN101653420A (en) * | 2009-09-08 | 2010-02-24 | 中国科学院化学研究所 | Ultrasonically controlled-release target medicinal preparation and production method thereof |
| CN102083412A (en) * | 2008-04-25 | 2011-06-01 | 杰伊·N·沙皮拉 | Programmed-release, nanostructured biological construct for stimulating cellular engraftment for tissue regeneration |
| CN102112176A (en) * | 2008-07-23 | 2011-06-29 | 皇家飞利浦电子股份有限公司 | Ultrasound mediated drug delivery |
-
2013
- 2013-01-08 CN CN201310006275.1A patent/CN103055302B/en not_active Expired - Fee Related
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1544098A (en) * | 2003-11-12 | 2004-11-10 | 邹金生 | Highly effective NGF adhesive and its preparation and using method |
| CN1946352A (en) * | 2004-04-09 | 2007-04-11 | 狄科特康坦特公司 | Methods and articles for the delivery of medicaments to the eye for the treatment of posterior segment diseases |
| CN101020061A (en) * | 2007-03-06 | 2007-08-22 | 西安交通大学 | Prepn process of ultrasonically controlled releasing micro gel containing medicine |
| CN101536984A (en) * | 2008-03-20 | 2009-09-23 | 江苏先声药物研究有限公司 | Injection-use recombinant human Endostatin porous sustained-release microsphere and preparation method thereof |
| CN102083412A (en) * | 2008-04-25 | 2011-06-01 | 杰伊·N·沙皮拉 | Programmed-release, nanostructured biological construct for stimulating cellular engraftment for tissue regeneration |
| CN102112176A (en) * | 2008-07-23 | 2011-06-29 | 皇家飞利浦电子股份有限公司 | Ultrasound mediated drug delivery |
| CN101653420A (en) * | 2009-09-08 | 2010-02-24 | 中国科学院化学研究所 | Ultrasonically controlled-release target medicinal preparation and production method thereof |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110917332A (en) * | 2019-12-12 | 2020-03-27 | 南通大禹生物技术有限公司 | Endodu in-vivo controlled-release medicine and preparation method and application thereof |
| CN110917332B (en) * | 2019-12-12 | 2023-11-07 | 南通大禹生物技术有限公司 | Endu in-vivo controlled release medicine and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103055302B (en) | 2014-05-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Delaney et al. | Breast cancer brain metastasis response to radiation after microbubble oxygen delivery in a murine model | |
| CN107875140A (en) | A kind of double targeted drug delivery systems and its application in oncotherapy preparation is prepared | |
| US20090053208A1 (en) | Methods and Systems for Improving Tissue Perfusion | |
| CN101653420A (en) | Ultrasonically controlled-release target medicinal preparation and production method thereof | |
| Moore et al. | Polymeric biomaterial scaffolds for tumoricidal stem cell glioblastoma therapy | |
| CN103990185B (en) | A kind of carrageenan and gelatin microsphere embolization agent and preparation method thereof | |
| CN103110567B (en) | The Preparation method and use of the administration nano-drug administration system of a kind of year tanshinone ⅡA | |
| Zhou et al. | Platelet-targeted microbubbles inhibit re-occlusion after thrombolysis with transcutaneous ultrasound and microbubbles | |
| CN103055302B (en) | Ultrasonic biological effect mediated recombinant human endostatin controlled release preparation | |
| CN104224750A (en) | Cabazitaxel albumin nanoparticle preparation for injection and preparation method thereof | |
| Ng et al. | Effect of acoustic cluster therapy (ACT®) combined with chemotherapy in a patient-derived xenograft mouse model of pancreatic cancer | |
| CN103877625B (en) | PLGA (poly(lactic-co-glycolic acid))/bletilla composite microsphere for embolism and preparation method thereof | |
| CN105582540A (en) | Brain glioma targeting drug delivery system by aiming at fatty acid-binding protein acceptor | |
| CN116687833A (en) | Soluble microneedle containing antibody-coupled albumin taxol nano-drug and preparation method and application thereof | |
| CN101972493B (en) | Visualized iodized oil-5-fluorouracil loaded polylactic acid microsphere preparation and preparation method thereof | |
| CN103735504A (en) | Irinotecan nano lipid binding preparation and preparation method thereof | |
| CN106974896A (en) | A kind of antitumor agent containing hydrophobicity chemotherapeutics nano particle and Fibrin Glue | |
| CN104324032B (en) | Antituberculotics three compound recipe microsphere vascular targeting thromboembolism slow releasing agent and its production and use | |
| CN109364253A (en) | One kind is for improving infiltrative nanoparticle of tumor tissues and its preparation method and application | |
| CN102727893B (en) | Recombinant human endostatin liposome microbubble and application thereof in preparing medicament for inhibiting tumor angiogenesis under ultrasound mediation | |
| CN105582539B (en) | It is a kind of for glioma cell and its rely on blood vessel drug delivery system | |
| CN102670611B (en) | Vascular targeting embolism sustained release agent of triple compound microsphere for antituberculosis drug, preparation method and applications thereof | |
| RU2257219C1 (en) | Method for treating noninvasive cancer of urinary bladder | |
| CN114177344A (en) | Liquid embolic agent and preparation method and application thereof | |
| CN105641704B (en) | A kind of production bubble polylactic-co-glycolic acid effervesce drug and preparation method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140514 Termination date: 20150108 |
|
| EXPY | Termination of patent right or utility model |