CN109876008A - A kind of drug and its preparation method and application for oncotherapy - Google Patents
A kind of drug and its preparation method and application for oncotherapy Download PDFInfo
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Abstract
A kind of drug and its preparation method and application for oncotherapy; the present invention relates to a kind of drugs; the problem of being discharged for the targeting delivery of hydrogen and controllability; an object of the present invention is to provide a kind of application in treatment tumour of metal boride and for the drug of oncotherapy, and the drug includes the protective agent of nano metal boride and surface modification.New Two Dimensional nanometer sheet material prepared by the present invention has the advantages that 1) the resulting MBNs two-dimension nano materials of the present invention have good biological safety；2) the resulting MBNs two-dimension nano materials of the present invention have preferable sour responsiveness；It 3) can be with radiotherapy and chemotherapy medicine combination therapy gastric cancer.4) normal cell and tissue are not influenced；5) hydrogen that MBNs@PVP discharges under stomach acidic environment has the effect of attenuation synergistic to chemicotherapy.
Description
Technical field
The present invention relates to a kind of drug, especially a kind of drug and its preparation method and application for oncotherapy.
Background technique
Chemicotherapy is clinical treatment cancer, especially the main method of middle and terminal cancer, is a kind of effective systemic control
Treatment means.However, traditional radiotherapy and chemotherapy medicine and administration mode, there is radiotherapy and chemotherapy medicines without specific recognition, and easy damaged is just
Normal cell and tissue, or even cause systemic toxicity, and tumour cell intake drug efficiency is low, repetitively administered easily causes tumour thin
Born of the same parents generate the disadvantages of drug resistance.
Gases for Treating is main using with therapeutic gas (hydrogen H as a kind of emerging oncotherapy technology2, one
Nitrogen oxide NO, carbon monoxide CO, hydrogen sulfide H2S etc.) replace or assists traditional chemicotherapy, reduce the secondary work of poison of radiotherapy and chemotherapy medicine
With.Hydrogen colorless and odorless is nontoxic, and hydrogen molecule is a kind of effective antioxidant, and it is stronger virulent that alternative neutralizes oxidisability
Property active oxygen, such as hydroxy radical and nitrous acid anion, but to the weaker active oxygen of oxidisability without obvious effect.Studies have shown that
Take in hydrogen-rich water to the internal organs ischemical reperfusion injury such as the heart, brain, kidney, small intestine have significant protective effect, to aging, diabetes,
The eremacausises such as parkinsonism stress disease also has obvious therapeutic effect.Hydrogen can not only promote Apoptosis of Colon Cancer Cells, connection
It closes when chemotherapeutics 5 FU 5 fluorouracil uses with more synergistic effect, Toxicity of Kidney can also be alleviated, it is antitumor without influencing its
Effect.Hydrogen can alleviate liver cancer patient radiotherapy adverse reaction, significantly improve Quality of Life of Tumor Patients, without influencing radiotherapy effect
Fruit.
The administration mode of hydrogen is mainly the physiological saline etc. for inhaling hydrogen, drinking hydrogen water, injecting hydrogen at present, these methods or
It is difficult to reach depth focal zone or the controlled release of gas is inefficient, thus be directed to the targeting delivery and controllability of hydrogen
Release is the significant challenge that current hydrogen treating is faced.
Summary of the invention
In view of the problems of the existing technology, the cancer situation based on current rigorous and the following nano biological medical development
Trend, using hydrogen as a kind of rosy prospect of nontoxic Novel medical gas, the present invention provides utilize liquid phase
Ultrasonic stripping method prepares a kind of metal boride nanometer sheet with sour responsiveness release hydrogen as releasing hydrogen drug, it is by two dimension
The metal boride of nanometer chip architecture and the protective agent of surface modification are constituted, and are realized and are discharged hydrogen to acidic environment responsiveness.Together
When, this Nano medication is devised peroral dosage form in conjunction with suitable drug auxiliary material by the present invention, is combined for same radiotherapy and chemotherapy medicine and is controlled
It treats, realizes hydrogen to the synergism and attenuation of radiotherapy and chemotherapy medicine.
An object of the present invention is to provide a kind of application of metal boride in treatment tumour, and the tumour is to disappear
Change road tumour, gastric cancer and intestinal cancer especially in situ.
A kind of drug for oncotherapy, the drug include nano metal boride.
Preferably, the metal boride is two-dimensional nano chip architecture, the general 1-2nm of the thickness of two-dimensional nano piece, grain
Sub general diameter 200nm or so, safe dose 2.5g/kg.
Preferably, the metal boride is MgB2、MnB2、FeB2、CaB6、SrB6、BaB6、TiB2、 ZrB2、EuB6、
GdB6, AgB and its homologue.
Further, the drug further includes the protective agent for having surface modification, this Nano medication can be in acidic environment
Responsiveness discharges hydrogen, and the protective agent is a kind of compound that can be complexed with metal boride, including but not limited to
Ethylenediamine tetra-acetic acid and its esters, polyvinylpyrrolidone, polyethylene glycol, hyaluronic acid and its derivative, the metal boron
The mass ratio of compound and compound is (1:2)~(1:10).
In addition, the present invention also provides peroral dosage form is prepared using this Nano medication, combined radio chemotherapy carries out the connection of tumour
Treatment is closed, realizes hydrogen to the synergism and attenuation of chemicotherapy, current existing hydrogen treating is solved and easily spreads, loading
It is low, it is difficult to the technical issues of controlling.
The peroral dosage form be by metal boride with auxiliary material with biocompatibility such as PVP, agarose etc. with (10:
1)~(30:1) ratio is tabletted, and the tablet and radiotherapy and chemotherapy medicine are used in combination, and the radiotherapy and chemotherapy medicine is chemotherapeutics
Dox, Doxorubicin or mitomycin etc., wherein it is preferred that chemotherapeutics is Dox.
The present invention also provides a kind of process for preparing medicine for oncotherapy, the preparation method includes following step
It is rapid:
(1) metal boride powder material and protective agent (1:2) in mass ratio~(1:10) are dispersed in water, then again
(10:1)~(100:1) chemical etchant is added thereto；
(2) above-mentioned mixed solution is subjected to liquid phase removing with sonicator again, using 100W~150W Mechanical Crushing
20min~120min；
(3) broken solution is first centrifuged off unreacted bulky grain with 700rpm-1500rpm, then by supernatant
Continue to be centrifuged with 10000rpm~13000rpm, be washed with deionized water after centrifugation 2 times, dehydrated alcohol is washed 2 times, and product is true
Empty drying can obtain final product.
The chemical etchant includes two class etching agents: boron etching agent and metal etch agent, two kinds of etching agents rise respectively
To effect be etch metal boride boron layer and metal layer, by volume (10:1)~(100:1) be added.Boron etching agent
It is a kind of weak oxidant, including but not limited to hydrogen peroxide, oxygen, ozone.Metal etch agent is a kind of weak acid, including but not office
It is limited to acetic acid, carbonic acid, hypochlorous acid, citric acid, malic acid.
New Two Dimensional nanometer sheet material prepared by the present invention has the advantages that
1) the resulting MBNs two-dimension nano materials of the present invention have good biological safety；
2) the resulting MBNs two-dimension nano materials of the present invention have preferable sour responsiveness, and hydrogen can be discharged in acidic environment
Gas does not discharge hydrogen substantially under neutral and alkaline environment, and can control its hydrogen after protective agent and excipient substance package
Gas release time makes it match with the intestines and stomach metabolism time, maximizes drug effect；
3) the resulting MBNs@PVP two-dimensional nano piece drug of the present invention is compared for hydrogen treating inhales hydrogen, drinks hydrogen water, note
Hydrogen saline therapy is penetrated, stomach, controlled release hydrogen can be targeted by oral realize, and can combine with radiotherapy and chemotherapy medicine
Treat gastric cancer.
4) hydrogen that the resulting MBNs@PVP Nano medication of the present invention discharges under gastric acid environment is only harmful to tumour cell,
Normal cell and tissue are had little influence on, and also can extend survival with chemotherapy combined URIN Treatment tumour in animal level
Time.
5) MBNs@PVP Nano medication is taken orally into stomach and chemicotherapy and plays synergy therapeutic effect, body by the present invention
Reveal the effect that the hydrogen that MBNs@PVP discharges under stomach acidic environment has attenuation synergistic to chemicotherapy, on a cellular level
Hydrogen is demonstrated to the synergism and attenuation of chemicotherapy.
Detailed description of the invention
Fig. 1 shows TEM, the SEM image that the two-dimentional boronation magnesium Nano medication of hydrogen is released in acid response of the invention；
Fig. 2 shows XRD, Mapping images that the two-dimentional boronation magnesium Nano medication of hydrogen is released in acid response of the invention；
Fig. 3 shows acid response of the invention and releases the ultra-violet absorption spectrum of two-dimentional boronation magnesium Nano medication of hydrogen and infrared
Map.
Fig. 4 shows responsiveness of the two-dimentional boronation magnesium Nano medication to the PBS of different pH that hydrogen is released in acid response of the invention
Discharge H2Behavior；
Fig. 5 show acid response of the invention release hydrogen two-dimentional boronation magnesium Nano medication and its after carrying out PVP package
The behavior of responsiveness release H2 in simulation hydrochloric acid environment；
Fig. 6, which is shown, studies hydrogen respectively on normal cell and tumor cell levels to the attenuation of chemicotherapy in the present invention
Synergy test result figure；
Fig. 7, which is shown, studies hydrogen combined radio chemotherapy respectively to normal cell and tumour cell in different time in the present invention
ATP change of production；
Fig. 8 shows the two-dimentional boronation magnesium Nano medication that hydrogen is released in acid response in the present invention and chemoradiotherapy plus treatment to dynamic
Therapeutic effect figure in object level；
Fig. 9 shows the two-dimentional boronation magnesium Nano medication that hydrogen is released in acid response in the present invention and chemoradiotherapy plus treatment to small
The attenuating effects of mouse cardiac muscular tissue are evaluated；
Figure 10 shows the two-dimentional boronation magnesium Nano medication that hydrogen is released in acid response in the present invention and chemoradiotherapy plus treatment to small
The tissue slice images of the major organs (liver, spleen, lung, kidney) of mouse.
Specific embodiment
Below with reference to embodiment, the present invention is described in further detail and completely, but is not intended to limit the contents of the present invention.
Embodiment 1
The preparation of material: first by H2O(120ml)、MgB2(90mg), PEG (4000) (450mg) are mixed in a reservoir
It is even, it places on ice bath, and sequentially add H2O2(9ml) and HAc (180ul), then carries out glass breaking with Ultrasonic Cell Disruptor, surpasses
Sound is crushed instrument parameter setting are as follows: amplitude transformer 6, (intermittent time) 2.2s/ (working time) 6.6s, altogether ultrasound 60min.It is super
After the completion of sound is broken, it is dispensed into 50ml centrifuge tube at once and is centrifuged (12500r/s, 25min), and gained precipitating is washed two
All over (12500r/s, 25min), dehydrated alcohol is washed twice (12500r/s, 10min).Vacuum drying 5h will be finally precipitated, will be obtained
New Two Dimensional boronation magnesium nanometer sheet MBNs.
Fig. 1 is transmission electron microscope (TEM), scanning electron microscope (SEM) image of material, shows that the MBNs material of above-mentioned preparation is
The reticular structure that two-dimensional nano piece is stacked up, Fig. 2 are XRD, Mapping image of New Two Dimensional nano material, can be from figure
Two-dimension nano materials prepared by A contain MgB2、MgB4Two kinds of crystal phases, and scheme to contain in the Mapping image displaying material of B
Some elements mainly have Mg, B, O.Fig. 3 shows the ultra-violet absorption spectrum and its near infrared spectrum of MBNs, near infrared light
Spectrum it can be seen that show O should from the coating of PEG, formed on the surface of MBN borate borate (B-O-PEG) and
HO-B, such as in 2470 and 1165cm-1What emerging peak reflected.
Fig. 4 illustrates novel two nanometer sheet MBNs made from embodiment 1 and utilizes the probe of MB+Pt in different pH in vitro
PBS in detection its hydrogen release (hydrogen of generation makes MB+Pt probe fade), indicate MBNs pH value be respectively 7.4,
6.8, the hydrogen release conditions in 5.0,1.2 PBS, the results showed that, there is no hydrogen release in the PBS that pH value is 7.4, with
The reduction of pH value, hydrogen release time is also shorter and shorter, and hydrogen release time of the MBNs in simulation hydrochloric acid environment only has
2h or so.
Embodiment 2
External hydrogen combined chemotherapy drug DOX is detected to the toxicity and ATP yield effect situation of cancer cell and normal cell,
The cell chosen respectively is stomach cancer cell (BGC-823) and four kinds of normal cells (H9C2, LO2, mMSC, HEK-
293cells), and by every kind of cell it is divided into air group, air+DOX (1 μ g/mL, 0.5 μ g/mL, 0.25 μ g/mL, 0.125 μ
G/mL), hydrogen group, hydrogen+DOX (1 μ g/mL, 0.5 μ g/mL, 0.25 μ g/mL, 0.125 μ g/mL) group detect hydrogen to change
The synergism and attenuation of drug is treated, wherein hydrogen group, hydrogen+DOX (1 μ g/mL, 0.5 μ g/mL, 0.25 μ g/mL, 0.125 μ g/
ML) group uses the special hydrogen incubator culture for specifically containing 60%.Fig. 6 illustrate on a cellular level hydrogen to chemotherapeutic
The synergism and attenuation of object, the results showed that, DOX concentration is that (1 μ g/mL, 0.5 μ g/mL, 0.25 μ g/mL, 0.125 μ g/mL) is right
The extent of damage of cancer cell increases with the increase of DOX concentration, and also has identical injuring rule to normal cell, but after logical hydrogen
The damage of cancer cell is reinforced, is that the damage to normal cell weakens, it was demonstrated that hydrogen plays the role of attenuation synergistic to chemotherapeutics.
The cell ATP production history of different incubation time upon administration (3h, 6h, 12h, for 24 hours) is had detected simultaneously.Figure
7 be testing result figure, shows H2The ATP level of the normal cell of processing will fluctuate slightly around in normal value, but most of
In the case of be usually slightly above normal level.Damage of the DOX to DNA can also damage mitochondrial function, therefore DOX and H2It can cooperate with
The breathing (figure A) for limiting cancer cell, to generate the cytotoxicity of collaboration enhancing to cancer cell.On the other hand, H2To normal thin
The activation of born of the same parents' energy effectively overcomes DOX and loses to cellular energy caused by injury of mitochondria, keeps the energy of mMSC cell
Near normal level (figure B), H is reflected2Chondriosome protective and injury-resistance.H2To the line grain of liver cell and splenocyte
The case where protection of bulk damage is with mMSC cell within a certain period of time is similar (figure DE).But seem DOX to heart, liver and spleen
Dirty long-term injury of mitochondria is under present circumstances almost without recovery (figure CDE), it can be deduced that H2Protective effect and thin
Born of the same parents' type is highly relevant, and depends on cell to the DOX and H of various concentration2Sensibility.
Embodiment 3
The present embodiment is by demonstrating the oral dosage combination of novel nano drug of the invention on model of gastric carcinoma in situ
Intravenous injection Dox can achieve synergism and attenuation.
The specific production step of oral preparation first are as follows: the MBNs for preparing embodiment 1 and PVP (55000) are by 1:19's
Mass ratio ground and mixed is uniform, carries out tabletting (pressure size 8*104N or so), the material MBNs@finally taken orally
PVP.And according to hydrogen of the method detection MBNs@PVP for the probe for utilizing MB+Pt in embodiment 1 in simulation hydrochloric acid environment
Release conditions, Fig. 5 indicate that detection MBNs@PVP hydrogen of (PBS, pH=1.2) in simulation hydrochloric acid environment is released in aforementioned manners
To one's heart's content condition, the results showed that, the MBNs after PVP is wrapped up extends hydrogen release time (release hydrogen 8h or so completely), reaches
The purpose of sustained release hydrogen is arrived.
Since mouse can not directly swallow peroral dosage form, so mouse feed and Nano medication are used above-mentioned tabletting side
Method is prepared into oral type feed.Particularly: mouse feed is worn into powdery, and MBNs PVP according to the ratio of 5mg/4995mg
Example is pressed into diet shape, feeding mouse, the above-mentioned special feed of the daily feeding 5g of every mouse with tablet press machine after mixing；Chemotherapy
The specific administration mode of drug: selected chemotherapeutic level adriamycin (DOX), administration mode are intravenous injection, dosage 3mg/
Kg, injection in each three days is primary, continuously injects a course for the treatment of (three times).It is commented according to the survival condition of each group mouse and histology
Valence come judge treat gastric cancer result.
Fig. 8 show PBS in animal level, MBNs@PVP, DOX, MBNs@PVP+DOX group treatment effectiveness evaluation figure, figure
A compared the time-to-live length of each group, and the time-to-live longest of MBNs@PVP+DOX group, figure B and the haemocyte evaluation for scheming C can
With find mouse can DOX treat later period (after 29 or 36 days) voluntarily adjust WBC and RBC quantity to normal level, but
The synergy of MBN@PVP and DOX can significant acceleration leucocyte and red blood cell restore normal level, directly demonstrate MBNs@
PVP plays the role of attenuation synergistic to chemotherapeutics in the hydrogen of stomach acid response release, is cut by the heart between each group of Fig. 9
The H&E dyeing of piece and Masson, which dye available MBNs@PVP, can mitigate chemotherapeutics in the hydrogen of stomach acid response release
The myocardial fibrosis caused by heart.Figure 10 shows the Histological Study comparative situation between each tissue, it was demonstrated that material itself
Apparent toxic side effect is not caused to mouse normal tissue.
Claims (16)
1. a kind of application of metal boride in the drug for the treatment of tumour.
2. application as described in claim 1, which is characterized in that the tumour is tumor in digestive tract, preferably the digestion
Road tumour is gastric cancer and intestinal cancer in situ.
3. a kind of drug for oncotherapy, which is characterized in that the drug includes nano metal boride.
4. being used for the drug of oncotherapy as claimed in claim 3, which is characterized in that the metal boride is two wieners
Rice chip architecture.
5. being used for the drug of oncotherapy as claimed in claim 3, which is characterized in that the metal boride is MgB2、
MnB2、FeB2、CaB6、SrB6、BaB6、TiB2、ZrB2、EuB6、GdB6, AgB and its homologue.
6. being used for the drug of oncotherapy as claimed in claim 3, which is characterized in that the drug further includes having surface to repair
The protective agent of decorations.
7. being used for the drug of oncotherapy as claimed in claim 6, which is characterized in that the protective agent is can be with metal
The compound of boride complexing, the preferably compound are ethylenediamine tetra-acetic acid and its esters, polyvinylpyrrolidone, poly- second
Glycol, hyaluronic acid and its derivative.
8. being used for the drug of oncotherapy as claimed in claim 6, which is characterized in that the metal boride and compound
Mass ratio be (1:2)~(1:10).
9. as claimed in claim 3 be used for oncotherapy drug, which is characterized in that the drug further include have PVP and/
Or agarose.
10. being used for the drug of oncotherapy as claimed in claim 10, which is characterized in that the drug is peroral dosage form.
11. being used for the drug of oncotherapy as claimed in claim 10, which is characterized in that the drug and radiotherapy and chemotherapy medicine
It is used in combination.
12. being used for the drug of oncotherapy as claimed in claim 11, which is characterized in that the radiotherapy and chemotherapy medicine is chemotherapy
Drug Dox, Doxorubicin or mitomycin.
13. a kind of process for preparing medicine for oncotherapy, which is characterized in that the preparation method includes following step
It is rapid:
(1) metal diboride powder and protective agent (1:2) in mass ratio~(1:10) are dispersed in water, are then added thereto again
Enter (10:1)~(100:1) chemical etchant；
(2) above-mentioned mixed solution is subjected to liquid phase removing with sonicator again；
(3) broken solution is centrifuged off unreacted bulky grain, then by supernatant continue with 120000rpm~
13000rpm centrifugation, particle washs after centrifugation, and product vacuum drying can be obtained final product.
14. being used for the process for preparing medicine of oncotherapy as claimed in claim 13, which is characterized in that the chemical etching
Agent includes boron etching agent and metal etch agent.
15. being used for the process for preparing medicine of oncotherapy as claimed in claim 14, which is characterized in that boron etching agent is a kind of
Weak oxidant, including but not limited to hydrogen peroxide, oxygen, ozone.
16. being used for the process for preparing medicine of oncotherapy as claimed in claim 14, which is characterized in that the metal etch
Agent is a kind of weak acid, including but not limited to acetic acid, carbonic acid, hypochlorous acid, citric acid, malic acid.
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| CN111743881A (en) * | 2020-07-07 | 2020-10-09 | 深圳大学 | Acid-responsive hydrogen-releasing drugs, preparation method thereof and application thereof in treatment of glycometabolism disorders and/or lipid metabolism disorders |
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| CN114053441A (en) * | 2021-10-09 | 2022-02-18 | 中国辐射防护研究院 | Radioactive embolism microball containing nano silicon and its preparing method, composition and use |
| CN114053599A (en) * | 2021-10-09 | 2022-02-18 | 中国辐射防护研究院 | Medicine box for tumor immunotherapy |
| CN115920110A (en) * | 2023-01-03 | 2023-04-07 | 大连理工大学 | Preparation method of nano metal boride composite material |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115920110A (en) * | 2023-01-03 | 2023-04-07 | 大连理工大学 | Preparation method of nano metal boride composite material |
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